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PubMed2006

Important experimental parameters for determining infection rates in arthropod vectors using pool screening approaches.

Measuring transmission of a vector-borne infection is essential to understanding infection dynamics. When infection prevalence in the vector population is low, transmission is often measured by pool screening (also referred to as group testing). Several investigators have developed statistical methods to recover infection prevalence estimates from pool screen data. These are based on models that contain certain assumptions, and a pool screening approach must be designed to take these into account if accurate estimates of infection prevalence are to be obtained. Here we describe these assumptions and discuss appropriate sampling protocols. The sources of error inherent in pool screening are described, and we show that, under most conditions in which one would want to use group testing, most of the error results from sampling and not the pooling process. Issues involved in developing a sampling protocol, including the total number of insects to be screened and optimal pool size, are explored. The meaning of confidence intervals associated with prevalence estimates and the appropriate interpretation of these intervals are discussed.

Africa, Western↗
PubMed1987

Modification and evaluation of a multichannel blood cell counting system for blood analysis in veterinary hematology.

A multichannel, semiautomated, blood cell counting system (Coulter Counter Model S550) was modified for use in veterinary hematology by increasing both the erythrocyte and leukocyte aperture currents to 225 V and 195 V, respectively, followed by calibration with human blood. It was evaluated by use of 350 samples from dogs, cats, horses, and cows. Values for leukocyte count, erythrocyte count, mean corpuscular volume, and hematocrit generated by the S550 were compared with values generated by an automated multichannel counter with histogram capability and other reference procedures when appropriate. Mean differences for values between S550 and reference values were less than calibration tolerance limits for the instrument. Correlation coefficients were excellent for all values of each species. To assess behavior of leukocytes of the different species with respect to the counting threshold, leukocyte size distribution histograms were generated for all samples analyzed on the S550. Means for mean leukocyte volumes in diluent and lysing reagents were 55.5, 56.6, 67.4, and 72.8 fl for dogs, cats, horses, and cows, respectively. Canine leukocyte counts, because of small leukocyte size, were an average of 14% less for 5 samples analyzed on the unmodified instrument, compared with analysis after increasing the leukocyte aperture current. Leukocyte threshold failures attributable to interfering particles, resulting in falsely high counts, were recognized in 14%, 10%, 8% and 0% of feline, bovine, canine, and equine samples, respectively. The magnitude of error in these samples averaged 5% for cows and dogs, but was considered not important. However, leukocyte counts of feline samples in this group averaged 44% falsely high.

Animals↗
PubMed1986

Evaluation of "HemoCue," a new device for determining hemoglobin.

Hemoglobinometry according to the International Committee of Standardization in Hematology (ICSH) suffers from imprecision related to high sample dilution and from potential errors owing to sample turbidity. We have evaluated a new instrument, "HemoCue," that measures hemoglobin at two wavelengths as azide methemoglobin, without dilution. The HemoCue method is superior to the ICSH method: by correction for turbidity, it avoids false hemoglobin readings that may arise from hyperlipemia or some large M-component of the immunoglobulin M class. We find the equipment suitable for use in outpatient units.

Evaluation Studies as Topic↗
PubMed2006

Assessment of the apparent intra- and inter-sample variability in the collagen fibril diameter in the posterior corneal stroma of rabbits. A transmission electron microscopy study.

PURPOSE: To assess the natural variability in the diameter of small collagen fibrils in the lamellae of corneal stroma of healthy young rabbits. MATERIALS AND METHODS: The corneas of 6 young adult female New Zealand White rabbits (2.1 kg) were prepared for transmission electron microscopy (TEM) by fixation with a cacodylate-buffered 2% glutaraldehyde (pH 7.2-7.4, 320-340 mosm/kg). The corneas were embedded in Epon-Araldite, thin sections prepared from the central region of the posterior corneal stroma and stained with acidic uranyl acetate followed by acidic lead citrate. High magnification (x 33,000) micrographs were taken, and fibril diameters (FDs) measured at a final magnification of x 275,000 to a resolution of 2 nm. RESULTS: Assessment of sampling-related errors indicated that the average diameter of the fibrils within any particular micrograph could be estimated to within 1% or better by measures of 100 fibrils. Assessments of the intra-sample variance (6 micrographs taken from the same cornea) indicated a group mean FD of 32.4 +/- 3.6 nm, whilst the inter-sample variance (6 micrographs taken from 6 different corneas) yielded an average of 33.1 +/- 4.5 nm (n = 100 fibrils/micrograph, +/-SD). However, group-averaged data sets of FDs, while unimodal, were not normally distributed, and cumulative averaging indicated a fixed range of FDs across the data sets. CONCLUSIONS: Intra- and inter-sample variability in collagen FDs is very similar, but the analysis indicates that the collagen fibrils are not homogeneous and that closely adjacent lamellae can have subtle differences in average FD.

Animals↗
PubMed1987

Comparison of measurement of glomerular filtration rate by single sample, plasma disappearance slope/intercept and other methods.

Various in vitro methods employing radioactive indicators have been presented for the determination of glomerular filtration rate (GFR) including slope/intercept (S/I) and single sample (SS) methods. The S/I technique utilizes the analysis of the slope of a certain segment of the plasma disappearance curve of a suitable radioactive indicator and its intercept on the y axis. The SS method is based on the theoretical volumes of distribution derived from a single plasma concentration at some specific time after intravenous injection of the indicator. Using GFRs estimated from the compartmental analysis of the entire plasma disappearance curve as a reference standard, we have compared the errors of GFR estimation calculated from 12 S/I methods, 3 SS methods and 2 techniques in which original formulae were derived. The errors of the SS method of Tauxe et al. gave the lowest standard error of estimate (Sy X x) of all the methods. The S/I methods gave lowest errors when sampling times were taken between 60 min and 240 min. Almost all the errors observed here were significantly less than those observed using scintillation camera techniques. The SS method seems to be the method of choice for estimation of GFR by single injection techniques. The use of the scintillation camera in conjunction with the SS technique would provide a useful right to left ratio so that individual kidney GFR could be calculated.

Adolescent↗
PubMed2005

Report on the 12th International Society of Blood Transfusion platelet immunology workshop.

BACKGROUND AND OBJECTIVES: The aims of the 12th International Society of Blood Transfusion (ISBT) Platelet Immunology Workshop were to evaluate the proficiency of molecular human platelet antigen (HPA) genotyping and detection of platelet antibodies of unusual specificity or reactivity, to assess whether quantification of anti-HPA-1a is practicable, and to determine the variability of reagents and components used in the monoclonal antibody immobilization of platelet antigens assay (MAIPA). MATERIALS AND METHODS: Forty participants from 23 countries were sent 10 samples for DNA typing, five samples for antibody detection, a freeze-dried anti-HPA-1a standard, three samples for anti-HPA-1a quantification and a MAIPA method questionnaire. RESULTS: The detection and identification of HPA antibodies varied from 2.7 to 95% of participants. The number of HPA genotyping errors per sample ranged from 0 to 3.96% per HPA loci. The majority of laboratories were able to assign an arbitrary number of units/ml of anti-HPA-1a activity to the unknown samples. The MAIPA questionnaire indicated a wide variation among participants, both in method and in reagents used. CONCLUSIONS: The results obtained from this workshop highlighted deficiencies in testing regimes and identified a need for internationally available reference materials.

Antibodies, Monoclonal↗
PubMed1998

Glutathione measurement in human plasma. Evaluation of sample collection, storage and derivatization conditions for analysis of dansyl derivatives by HPLC.

Literature values for human plasma GSH vary over 10-fold despite the use of apparently valid analytical procedures for GSH measurement. The purpose of this study was to develop a procedure to minimize error in sample collection, processing and storage that could contribute to such differences. HPLC with fluorescence detection of dansyl derivatives was used for quantification. The results show that collection of blood with a butterfly needle and syringe reduces overestimation due to limited hemolysis and that use of a preservation solution designed to inhibit autooxidation and enzymatic degradation allows quantitative recovery of both GSH and GSSG. Stability tests showed that non-derivatized samples were stable for at least 2 months at - 80 degrees while dansyl derivatives were stable in the dark at 0-4 degrees for 12 months. Results from 59 healthy individuals (20-43 years) provided a mean (+/-1 SD) GSH value of 2.09+/-1.14 micromolar.

Blood Specimen Collection↗
PubMed2001

Objectives, concept and design of the CEEM soil project.

Within the member countries of the European Union, different strategies and guidelines for the sampling and the preparation of soils are used to investigate soil contamination. As a consequence, there is reasonable doubt that the results gained by applying these different strategies and methods are comparable. In general, the relative contribution of sampling and sample preparation to the total uncertainty in soil contamination studies is a subject of speculation than of knowledge. For this purpose, the first European soil sampling inter-comparison test was organised as a project under the Standards, Measurement and Testing Programme of the European Commission. The aim of this project was to analyse and to compare the existing guidelines, the degree and the state of standardisation in this field in the European countries, and to compare their results when applied on the same test area under comparable conditions by representative sampling teams. The results of the project in general demonstrate that sampling and sample preparation errors reach about the same order of magnitude as errors caused in the chemical analysis. However, there are great differences depending on the elements investigated and their concentration ranges, as well as on pedological and land-use conditions in the test area. Consequently, sampling and sample preparation require the same attentiveness and equivalent measures for quality assurance (QA) and quality control (QC) as the chemical analysis, which has not been the case up to now. Insufficient comparability of the results is illustrated by the fact that different participants arrive at different conclusions for up to 61% of tested soil quality criteria (e.g. threshold levels). The methods and results of the project are presented in detail in the six following papers within this volume. The following 13 papers present some key studies and specific contributions of the participants, as well as synoptical papers of the invited experts in standardisation and accreditation.

Environmental Monitoring↗
PubMed2000

Evaluation of the Roche Diagnostics LightCycler-Apo B 3500 Mutation Detection Kit.

Familial defective apolipoprotein (apo) B-100 is an autosomal codominant disorder associated with hypercholesterolemia and an increased risk of coronary artery disease. Two independent mutations affecting the codon 3500 (Arg3500-->Gln and Arg3500-->Trp) have been shown to cause ligand-defective apo B-100. Identification of carriers of these mutations is an important step in the risk stratification of individuals and families with hypercholesterolemia. We evaluated a homogeneous assay for detection of mutations at codon 3500 that combines rapid-cycle PCR with allele-specific fluorescent probe melting profiles for product genotyping. This single-tube analysis is performed on the LightCycler, a microvolume fluorimeter integrated with a thermal cycler. Continuous acquisition of fluorescence data during a melting curve analysis at completion of PCR allows the detection of mutations, as loss of fluorescence occurs in an allele-specific manner. By plotting melting peaks, the three apo B-100 alleles were readily distinguishable. Using this method, genotyping of 32 samples is completed within 40 min without the need for any post-PCR sample manipulation, thereby eliminating the risks of end-product contamination and sample tracking errors. The specific detection of mutations at codon 3500 of the apo B gene on the LightCycler is a rapid and reliable method that is ideally suitable for typing both small and large numbers of samples.

Apolipoprotein B-100↗
PubMed1987

Articulation error migration: a comparison of single word and connected speech samples.

In this study frequency and type of articulation error, including error migration, were compared between single word production and connected speech samples when vocabulary was held constant. Spontaneous single word and connected speech samples were obtained from 20 articulation disordered children ages 5-4 to 12-8 years. There were significantly more errors in connected speech samples than single word utterances. The difference between samples, i.e., error migration, is discussed, as is the predictive value for speech-language clinicians.

Articulation Disorders↗
PubMed2005

Loss of heterozygosity of APC and CDH1 genes in laryngeal squamous cell carcinoma.

The molecular mechanisms involved in the development and progression of laryngeal cancer, specifically squamous cell carcinoma, still need further investigation and elucidation. Twenty-two laryngeal squamous cell carcinomas were analyzed in our study regarding genetic changes of two tumor suppressor genes: Adenomatous polyposis coli (APC) and E-cadherin (CDH1). APC gene instability was tested by polymerase chain reaction (PCR)/loss of heterozygosity (LOH) using the restriction fragment length polymorphism (RFLP) method. The samples were also screened for mutations using the heteroduplex method. E-cadherin gene was analyzed by PCR amplification of tetranucleotide marker (D16S752) linked to E-cadherin gene. The results of our analysis showed three samples with LOH of the APC gene out of 15 heterozygous patients (20%). Only one LOH of the CDH1 gene (5.5%) out of 18 heterozygous patients was discovered. D16S752 marker did not reveal any replication error-positive samples. There were six samples showing heteroduplexes (33%) encompassed in APC's exon 11. Altogether, nine samples (41%) showed alterations of the APC gene. Our results suggest that alterations of APC gene may have a role in squamous cell carcinoma development. Detected LOH of the E-cadherin gene indicates that genetic changes of this gene are not very frequent, but that other components of the wnt signaling cascade may also be involved.

Adult↗
PubMed1976

The effect of measurement error on differences in hospital expenditures.

Differences in expenditures for medical care among various subgroups in the population according to age, income, residence, and ethnicity have important public policy implications. However, few efforts have been made to assess the extent to which these apparent differences among subgroups observed in social surveys may be influenced by measurement error. This paper provides a framework for examining the impact of various kinds of error including random error and biases. The framework is applied to data on hospital expenditures from a national survey of health services expenditure which includes in addition to consumer reports, verifying information from hospitals, physicians, and third-party payers who provided services to persons in the sample. Differential error according to demographic and social characteristics is estimated. The impact of such differential error on conclusions about the distribution of health resources is discussed.

Age Factors↗
PubMedDate not supplied

Role of exposure databases in epidemiology.

At present, exposure databases record data primarily for regulatory purposes; they have not focused on serving the needs of epidemiologists or public health. However, the modification of exposure databases could facilitate their use in epidemiology. Characteristics necessary to enhance the use of all databases include easy access by users; documentation of methods, sampling bias, error, and inconsistences; widespread coverage in time and space; and methods and measures for estimating exposure of individuals as well as populations. Also needed are exposure scenarios and models to estimate exposures for geographic areas and time intervals not currently sampled. Multidisciplinary teams are needed to examine current databases, to review strategies for improving data collection, and to suggest and help implement appropriate changes. A long-term goal is to develop and validate data from exposure scenarios and models using data on the relationship of exposure to doses measured in humans.

Databases, Factual↗
PubMedDate not supplied

Evaluating the results of mail survey research.

OBJECTIVE: To present overall guidance enabling the reader to critically evaluate survey research conducted via mail questionnaire. Also considered are the sources of error that may operate during the conduct of survey research and the threats they pose to both the internal rigor of the study and the generalizability of the results. STUDY SELECTION: Not applicable. DATA EXTRACTION: Not applicable. DATA SYNTHESIS: Not applicable. CONCLUSION: Survey research has been used so commonly and for such a wide range of purposes in a large number of fields, that some people may mistakenly conclude that surveys are easy to conduct. Survey research involves much more than generating a set of questions to ask a group of subjects. To draw valid conclusions about the topic of a survey, the pharmacist must be prepared to evaluate the results in light of how well the investigators approached four sources of error--coverage, sampling, measurement, and nonresponse. The evaluation of survey administration procedures primarily concerns determining whether or not a pilot study was conducted, whether proper statistical analyses were conducted, and whether multiple mailing procedures were used. Failure to account for these sources of error, or inappropriate survey administration, could seriously affect both the generalizability and validity of the results.

Data Interpretation, Statistical↗
PubMed1987

Cisternal cerebrospinal fluid concentrations of morphine following intrathecal and epidural administration in the baboon.

Eleven baboons, anaesthetised with ketamine, had catheters introduced into the cisterna magna. Morphine was injected at lumbar level intrathecally in six and epidurally in five. Cisternal CSF was sampled hourly and the morphine concentration measured using a high pressure liquid chromatograph. In two cases following intrathecal injection peaks of 180 ng/ml at 4 hours, and 2,200 ng/ml at 3 hours were detected. In the latter case there was associated error in sampling therefore this baboon had a repeat injection four weeks later. The maximum level of morphine obtained then was 139 ng/ml at 4 hours. In the epidural group peaks of 113 ng/ml and 53 ng/ml at 1 hour were measured in 2 baboons and 27 ng/ml at 6 hours in a third. In all six other baboons following either procedure no morphine was detected in the cisterna. We conclude that morphine injected either intrathecally or epidurally in primates does migrate centrally in varying quantities. This finding would seem to have some bearing on the unpredictability of reported episodes of respiratory depression following intraspinal morphine.

Animals↗
PubMed2001

Bias assessment of current technologies used for the determination of low levels of moisture in mineral oil samples.

The problem in the current debate on the accuracy of Karl Fischer (KF) titrations lies in the fact that coulometry is being compared to volumetry on mineral oil samples for which the true moisture content is unknown. To clarify this point, dehydrated oil samples equilibrated under known temperature and relative humidity conditions and equilibrated oil samples containing known amounts of added moisture were used to assess the accuracy of the determinations. In addition, the measurements were extended to other techniques given that it is unlikely that they would be affected by the same phenomenon causing the KF systematic errors. The samples sent to different laboratories were analyzed by headspace/capillary gas chromatography, gas-phase H2 sensor, oil-phase or gas-phase RH sensors, KF coulometric titration with direct or indirect injection, and KF volumetric titration using a standard or NIST modified procedure. The laboratory comparison showed that with the exception of 4 techniques out of 10 that were tested, the measurements gave results in the expected concentration range. Considering the exceptions, two techniques based on volumetric titration yielded results tainted with an important positive bias for both sample types. This bias, tentatively associated with the high iodine end point concentration used by these systems, was estimated at approximately 22 ppm under the conditions applied by NIST. On the other hand, the two RH sensors showed a marked tendency to underestimate the value of the samples containing high moisture content. In this case, a loss of analyte through wall adsorption during the time required to achieve steady-state conditions in the measuring chamber seems to be at the origin of the negative biases.

Journal Article↗