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Interference by nonsteroidal anti-inflammatory drugs in EMIT and TDx assays for drugs of abuse.

Fourteen nonsteroidal anti-inflammatory drugs were evaluated for interference in EMIT and TDx assays for drugs of abuse. Only tolmetin demonstrated significant interferences in the EMIT assay. Urine samples that contained high concentrations of tolmetin (1800 mg/L) had characteristic high molar absorptivity at the wavelength used in EMIT assays (340 nm). Consequently, EMIT analysis of samples resulted in instrument error alarms on a Hitachi 704 instrument and depressed milliabsorbance values (delta A) relative to calibrators. Similar results were obtained with urine samples collected from an arthritic patient after the administration of 200 and 400 mg of tolmetin. When tolmetin samples were mixed with drugs of abuse, depressed delta A values were noted in all assays. Samples containing opiates and cannabinoids tested negative, and instrument error alarms were produced with samples that contained amphetamines. A gas chromatographic-mass spectrometric (GC-MS) assay for benzoylecgonine in the presence of tolmetin was successful, and no interferences were noted. Similar interferences by tolmetin were not observed in TDx assays, probably because of the different wavelength (525 nm) used in this assay. However, a potential for false-positive results in the TDx benzodiazepine assay was noted for urine samples containing high concentrations of fenoprofen, flurbiprofen, indomethacin, ketoprofen, and tolmetin. Generally, it was concluded that the presence of tolmetin in urine samples could lead to the production of unacceptable results by the EMIT assay for drugs of abuse. However, TDx and GC-MS assays were useful alternatives for the analysis of urine samples suspected of containing tolmetin.

Anti-Inflammatory Agents, Non-Steroidal↗

Exact mass measurement on an electrospray ionization time-of-flight mass spectrometer: error distribution and selective averaging.

An automated, accurate and reliable way of acquiring and processing flow injection data for exact mass measurement using a bench-top electrospray ionization time-of-flight (ESI-TOF) mass spectrometer is described. Using Visual Basic programs, individual scans were selected objectively with restrictions on ion counts per second for both the compound of interest and the mass reference peaks. The selected "good scans" were then subjected to two different data-processing schemes ("combine-then-center" and "center-then-average"), and the results were compared at various ion count limit settings. It was found that, in general, the average of mass values from individual scans is more accurate than the centroid mass value of the combined (same) scans. In order to acquire a large number of good scans in one injection (to increase the sampling size for statistically valid averaging), an on-line dilution chamber was added to slow down the typically rapid mass chromatographic peak decay in flow-injection analysis. This simple addition worked well in automation without the need for manual sample dilution. In addition, by dissolving the reference compound directly into the mobile phase, manual syringe filling can be eliminated. Twenty-seven samples were analyzed with the new acquisition and process routines in positive electrospray ionization mode. For the best method found, the percentage of samples with RMS error less than 5 ppm was 100% with repetitive injection data (6 injections per sample), and 95% with single injection data. Afterwards, 31 other test samples were run (with MW ranging from 310 to 3493 Da, 21 samples in ESI+ and 10 in ESI- mode) and processed with similar parameters and 100% of them were mass-calculated to RMS error less than 5 ppm also.

Automation↗

PLS regression using real sample calibration for aluminum and iron determination in plant extracts.

Real samples were used for PLS model calibration and validation steps, showing that this approach can be of value in preventing deviations in the results caused by the matrix effects for the simultaneous spectrophotometric determination of aluminum and iron in plant extracts. One hundred UV-vis spectra, obtained from samples of the 1997 to 2000 International Plant-Analytical Exchange (IPE) program (The Netherlands), were used for model development, with ICP-AES aluminum and iron determinations as reference values for model calculation. The plant extracts were analyzed both by ICP-AES and by the PLS models developed in this work, using calibrations with both aqueous standard solutions and with real sample extracts. In addition, since the use of smaller calibration sets could be of value in reducing both the cost and the time of analysis, sets with fewer calibration samples were also investigated, with the help of the Kennard and Stone algorithm for sample selection. Comparison of the predictability of the best model obtained with each calibration set was made using the ratio of their relative root mean square error (%RMSEV) for samples in the validation set, for aluminum or iron determinations, and were compared against F-test tabulated values. For all the models developed with real samples, the differences in the %RMSEV values for the aluminum or iron determinations were found not to be statistically significant, at a confidence level of 95%. Although it was observed that the aluminum, but not the iron, determinations with the PLS 2 model prepared with aqueous standards tend to be slightly lower than the ICP-AES determinations, this model has a good global prediction ability, as observed through the correlation curves presented, and can be used for screening determinations or for other agricultural purposes.

Aluminum↗

Accurate nucleic acid concentrations by nuclear magnetic resonance.

Determination of the concentration of biochemical samples often yields values with uncertainties of 10-20% or more. This paper details a protocol for use with 500- to 600-MHz NMR spectrometers to measure approximately 1mM concentrations within +/-1-3% accuracy. With suitable precautions, all compounds have equal NMR "absorption coefficients" for protons. About 2mg of sample are needed for proteins and nucleic acids with MW=5000, although less accurate determinations could be made with smaller amounts. The technique utilizes standardized internal reference reagent compounds, cacodylic acid or 3-(trimethylsilyl)propionic-2,2,3,3-d(4) acid sodium salt. Spectra were signal-averaged using long interpulse delays so that integrals of nonexchangeable protons could be quantified relative to the reference standard. Accurate determinations require careful optimization of the homogeneity of the magnetic field and meticulous attention to sample preparation and spectral processing. The main source of error is usually the accuracy of micropipets. If sample preparation errors could be eliminated, the lower limit of accuracy with the current generation of NMR spectrometers is probably near 0.4%. However, this would require >99.5% sample purity. Methods are described to establish the concentration of the standards, and applications are illustrated with DNA mono- and oligonucleotides. Similar procedures should apply to proteins, polysaccharides, and other biomolecules, with about the same accuracy and precision.

Cacodylic Acid↗

Mass spectrometry of medicines. Quantitative determination by direct probe inlet system mass chromatography.

A fundamental study was performed to establish a method of quantitative analysis using mass chromatography by the direct inlet system. The samples were aspirin, phenacetin and caffeine, which are often used as cold medicines, and barbital, allobarbital, phenobarbital and phenytoin which are difficult to analyse by gas chromatography in their intact states. N-acetylsulfamine and ethyl p-aminobenzoate were used as internal standards. Direct inlet mass chromatography was performed by an on-line system of the Shimadzu LKB-9000 and the GC-MSPAC 300. The ratio of the cumulative ions of certain peaks of sample and the internal standard was studied. It was found that, whether the sample is a pure reagent or a mixture, the ratio of cumulative ions of a peak specific to the sample and of a selected peak of the internal standard is proportional to the sample size, the error being less than +/- 3.5% for aspirin, phenacetin and caffeine, and less than +/- 2.7% for barbital, allobarbital and phenobarbital. The same relationship was observed for the phenobarbital and phenytoin mixed in rat plasma, the error being less than +/- 2.0%. It can be concluded that this method is applicable to the quantiative determination of medicines in urine, body fluids and other biological samples.

Aspirin↗

Estimating distributions of numbers of organisms in food products.

Procedures of sampling and measurement contribute variability and uncertainty to exposure models that predict incidence and levels of organisms in food products. This paper focuses on methods that account for sampling and measurement error in fitting distributions of organisms in food products for use in exposure models for microbial risk assessment. Define y to be a measured density on a sample selected with stipulated probability from a population, and define x to be the "true" density for that sample. Designate the conditional distribution of y given the sample with "true" value x as g(y/x), and let F(x) be the unknown cumulative density of x. The distribution of the observed values y, h(y), can be expressed through the integral equation h(y) = integral of g(y/x) dF(x). Knowledge of g(y/x) and h(y) enables an estimate of the unknown distribution of the organism's F(x). In applications to risk assessment, use of continuous distributions described by a few parameters is desirable. Also desirable may be imputation or assignment of possible nonzero values for nondetect observations--those results below the limit of detection of the methodology--that may not be truly zero. This paper explores the use of the above formulation for estimating distributions of organisms used for microbial risk assessment and presents some simple examples.

Clostridium perfringens↗

Managing patients with identical names in the same ward.

PURPOSE: To review the experience of managing two patients with identical names in the same ward during a five-month period. DESIGN/METHODOLOGY/APPROACH: The records of the patients were reviewed to look for incorrect entries, errors in specimens sampling, administration of blood products and chemotherapy, and misplacement of clinical notes. Doctors and nurses involved were also invited to complete a questionnaire study to comment on the usefulness of the measures implemented for correct patient identification. A random sample of 60 patients was also selected to see if their full names were shared with other patients attending the same hospital. FINDINGS: Among the 1442 sheets of hospital records from the two patients, no errors pertaining to the clinical activities were found. However, 13 (0.9 per cent) sheets of the hospital records were misplaced. The 21 doctors and nurses participating in the questionnaire study gave positive support to all the additional measures implemented for safeguarding patient identification, of which the automated alerting feature in the electronic clinical management system received the highest scores. A total of 32 (53 per cent) of the 60 sampled patients shared a common full name with one to 101 other patients attending the same hospital. ORIGINALITY/VALUE: Patients with identical names staying in the same ward present a unique challenge to acute health-care settings. The situation is especially relevant in communities where most people's names are not unique. Specific guidelines and measures are needed to prevent patient misidentification. Errors in filing of patient notes and laboratory reports to the hospital record deserve further attention.

Adolescent↗

Some sources of error and their effect on Census statistics.

Often the reliability of survey data is examined only in relationship to sampling variances, excluding many other potential sources of error. If the sampling variance dominates the mean-square error, then few mistakes result by considering sampling variance only; however, if sampling variance is only a small part of the mean-square error, serious mistakes in inference could be made. The Bureau of the Census has developed a model describing the joint effect of sampling and nonsampling errors on census statistics. This article shows how a study of the components of error may lead to methods of improving the accuracy and reliability of survey data.

Demography↗

The Mine Safety and Health Administration's criterion threshold value policy increases miners' risk of pneumoconiosis.

BACKGROUND: The Mine Safety and Health Administration (MSHA) proposes to issue citations for non-compliance with the exposure limit for respirable coal mine dust when measured exposure exceeds the exposure limit with a "high degree of confidence." This criterion threshold value (CTV) is derived from the sampling and analytical error of the measurement method. CONCEPTUAL FRAMEWORK: This policy is based on a combination of statistical and legal reasoning: the one-tailed 95% confidence limit of the sampling method, the apparent principle of due process and a standard of proof analogous to "beyond a reasonable doubt." CRITIQUE: This policy raises the effective exposure limit, it is contrary to the precautionary principle, it is not a fair sharing of the burden of uncertainty, and it employs an inappropriate standard of proof. Its own advisory committee and NIOSH have advised against this policy. For longwall mining sections, it results in a failure to issue citations for approximately 36% of the measured values that exceed the statutory exposure limit. DISCUSSION: Citations for non-compliance with the respirable dust standard should be issued for any measure exposure that exceeds the exposure limit.

Dust↗

Quantitative evaluation of the CEEM soil sampling intercomparison.

The aim of the CEEM soil project was to compare and to test the soil sampling and sample preparation guidelines used in the member states of the European Union and Switzerland for investigations of background and large-scale contamination of soils, soil monitoring and environmental risk assessments. The results of the comparative evaluation of the sampling guidelines demonstrated that, in soil contamination studies carried out with different sampling strategies and methods, comparable results can hardly be expected. Therefore, a reference database (RDB) was established by the organisers, which acted as a basis for the quantitative comparison of the participants' results. The detected deviations were related to the methodological details of the individual strategies. The comparative evaluation concept consisted of three steps: The first step was a comparison of the participants' samples (which were both centrally and individually analysed) between each other, as well as with the reference data base (RDB) and some given soil quality standards on the level of concentrations present. The comparison was made using the example of the metals cadmium, copper, lead and zinc. As a second step, the absolute and relative deviations between the reference database and the participants' results (both centrally analysed under repeatability conditions) were calculated. The comparability of the samples with the RDB was categorised on four levels. Methods of exploratory statistical analysis were applied to estimate the differential method bias among the participants. The levels of error caused by sampling and sample preparation were compared with those caused by the analytical procedures. As a third step, the methodological profiles of the participants were compiled to concisely describe the different procedures used. They were related to the results to find out the main factors leading to their incomparability. The outcome of this evaluation process was a list of strategies and methods, which are problematic with respect to comparability, and should be standardised and/or specified in order to arrive at representative and comparable results in soil contamination studies throughout Europe. Pre-normative recommendations for harmonising European soil sampling guidelines and standard operating procedures have been outlined in Wagner G, Desules A, Muntau H, Theocharopoulos S. Comparative Evaluation of European Methods for Sampling and Sample Preparation of Soils for Inorganic Analysis (CEEM Soil). Final Report of the Contract SMT4-CT96-2085, Sci Total Environ 2001;264:181-186. Wagner G, Desaules A, Munatu H. Theocharopolous S, Quevauvaller Ph. Suggestions for harmonising sampling and sample pre-treatment procedures and improving quality assurance in pre-analytical steps of soil contamination studies. Paper 1.7 Sci Total Environ 2001b;264:103-118.

Environmental Monitoring↗

Nonparametric AUC estimation in population studies with incomplete sampling: a Bayesian approach.

The estimation of the AUC in a population without frequent and/or fixed individual samplings is of interest because the number of plasma samples can often be limited due to technical, ethical and cost reasons. Non-linear mixed effect models can provide both population and individual estimates of AUC based on sparse sampling protocols; however, appropriate structural models for the description of the pharmacokinetics are required. Nonparametric solutions have also been proposed to estimate the population AUC and the associated error when particular sampling protocols are adopted. However, they do not estimate the individual AUCs and lack flexibility. Also a semiparametric method has been proposed for addressing the problem of sparse sampling in reasonably well designed studies. In this work, we propose and evaluate a nonparametric Bayesian scheme for AUC estimation in population studies with arbitrary sampling protocols. In the stochastic model representing the whole population, the individual plasma concentration curves and the "mean" population curve are described by random walk processes, allowing the application of the method to the reconstruction of any kind of "regular" curves. Population and individual AUC estimation are performed by numerically computing the posterior expectation through a Markov chain Monte Carlo algorithm.

Algorithms↗

Reproducibility and accuracy of measurements of free and total prostate-specific antigen in serum vs plasma after long-term storage at -20 degrees C.

BACKGROUND: Long-term frozen storage may alter the results of prostate-specific antigen (PSA) measurements, mainly because of degradation of free PSA (fPSA) in vitro. We compared the effects of long-term storage on fPSA, total PSA (tPSA), and complexed PSA (cPSA) in serum vs EDTA-plasma samples. METHODS: We measured fPSA and tPSA concentrations in matched pairs of archival serum and EDTA-plasma samples (stored frozen at -20 degrees C for 20 years) from a large population-based cohort in Malmö, Sweden. We also compared concentrations in age-matched men with those in samples not subjected to long-term storage, obtained from participants in a population-based study of prostate cancer screening in Göteborg, Sweden. These contemporary samples were handled according to standardized preanalytical and analytical protocols aimed at minimizing in vitro degradation. tPSA and fPSA measurements were performed with a commercial assay (Prostatus Dual Assay; Perkin-Elmer Life Sciences). RESULTS: Concentrations of tPSA and fPSA and calculated cPSA (tPSA - fPSA) in archival plasma were not significantly different from those in contemporary serum from age-matched men. In archival serum, however, random variability of fPSA was higher vs plasma than in contemporary samples, whereas systematic error of fPSA analyses was similarly small in archival and contemporary serum and plasma. CONCLUSIONS: Concentrations of tPSA and calculated cPSA were highly stable in plasma and serum samples subjected to long-term storage at -20 degrees C. Greater random variability, rather than a systematic decrease, may explain differences in fPSA analyses observed in archival serum.

Aged↗

Good-Enough RFLP Matcher (GERM) program.

A spreadsheet-based program (Good-Enough RFLP Matcher or GERM) is presented that matches unknown restriction fragment length polymorphism (RFLP) patterns of ectomycorrhizal fungi to a database of known ectomycorrhizal fungi. The program uses three simple methods to determine whether a sample matches a known: (1) Forward Matching: whether every band in the unknown is present in a known sample within a given error range; (2) Backward Matching: whether every band in the known sample is present in the unknown within a given error range; (3) Sum of Bands: whether the sum of all bands in the known and unknown are similar within a given error range. The program is available through the web page of this journal.

Database Management Systems↗

On corrected score approach for proportional hazards model with covariate measurement error.

In the presence of covariate measurement error with the proportional hazards model, several functional modeling methods have been proposed. These include the conditional score estimator (Tsiatis and Davidian, 2001, Biometrika 88, 447-458), the parametric correction estimator (Nakamura, 1992, Biometrics 48, 829-838), and the nonparametric correction estimator (Huang and Wang, 2000, Journal of the American Statistical Association 95, 1209-1219) in the order of weaker assumptions on the error. Although they are all consistent, each suffers from potential difficulties with small samples and substantial measurement error. In this article, upon noting that the conditional score and parametric correction estimators are asymptotically equivalent in the case of normal error, we investigate their relative finite sample performance and discover that the former is superior. This finding motivates a general refinement approach to parametric and nonparametric correction methods. The refined correction estimators are asymptotically equivalent to their standard counterparts, but have improved numerical properties and perform better when the standard estimates do not exist or are outliers. Simulation results and application to an HIV clinical trial are presented.

Acquired Immunodeficiency Syndrome↗

Unbiased estimation of relative reproductive success of different groups: evaluation and correction of bias caused by parentage assignment errors.

Parentage assignment is widely applied to studies on mating systems, population dynamics and natural selection. However, little is known about the consequence of assignment errors, especially when some parents are not sampled. We investigated the effects of two types of error in parentage assignment, failing to assign a true parent (type A) and assigning an untrue parent (type B), on an estimate of the relative reproductive success (RRS) of two groups of parents. Employing a mathematical approach, we found that (i) when all parents are sampled, minimizing either type A or type B error insures the minimum bias on RRS, and (ii) when a large number of parents is not sampled, type B error substantially biases the estimated RRS towards one. Interestingly, however, (iii) when all parents were sampled and both error rates were moderately high, type A error biased the estimated RRS even more than type B error. We propose new methods to obtain an unbiased estimate of RRS and the number of offspring whose parents are not sampled (zW(z)), by correcting the error effects. Applying them to genotypic data from steelhead trout (Oncorhynchus mykiss), we illustrated how to estimate and control the assignment errors. In the data, we observed up to a 30% assignment error and a strong trade-off between the two types of error, depending on the stringency of the assignment decision criterion. We show that our methods can efficiently estimate an unbiased RRS and zW(z) regardless of assignment method, and how to maximize the statistical power to detect a difference in reproductive success between groups.

Animals↗

Monitoring trends in under-5 mortality rates through national birth history surveys.

BACKGROUND: We assessed whether Demographic and Health Surveys (DHS), a large and high-quality source of under-5 mortality estimates in developing countries, would be able to detect reductions in under-5 mortality as established in global child health goals. METHODS AND RESULTS: Mortality estimates from 41 DHS conducted in African countries between 1986 and 2002, for the interval of 0-4 years preceding each survey (with a mean time lag of 2.5 years), were reviewed. The median relative error on national mortality rates was 4.4%. In multivariate regression, the relative error decreased with increasing sample size, increasing fertility rates, and increasing mortality rates. The error increased with the magnitude of the survey design effect, which resulted from cluster sampling. With levels of precision observed in previous surveys, reductions in all-cause under-5 mortality rates between two subsequent surveys of 15% or more would be detectable. The detection of smaller mortality reductions would require increases in sample size, from a current median of 7060 to over 20,000 women. Across the actual surveys conducted between 1986 and 2002, varying mortality trends were apparent at a national scale, but only around half of these were statistically significant. CONCLUSIONS: The interpretation of changes in under-5 mortality rates between subsequent surveys needs to take into account statistical significance. DHS birth history surveys with their present sampling design would be able to statistically confirm under-5 mortality reductions in African countries if true reductions were 15% or larger, and are highly relevant to tracking progress towards existing international child health targets.

Africa↗

[Prescription habits. Illustrated by a random sample].

With the object of illustrating prescribing habits and errors in prescriptions, a randomized sample of 1,000 prescriptions was employed from the prescriptions from a single day. The main result of the investigation is that, in general, a great proportion of errors occur as 61.7% of the prescriptions were incorrect. Errors concerning limiting of issue and indications of reasons for use are particularly noticeable. Only 48% of the drugs prescribed for which limitation of issue is compulsory and 32.4% of all preparations do not show the indication for use. The alterations in the circular on dispensing issued by the Danish Board of Health with the object of prompting doctors to write multiple prescriptions and always to give the indication for employment of the preparation concerned are thus far from fulfilled. Employment of telephonic prescriptions is still extensively employed as nearly 50% of all prescriptions are given by telephone and the intention of the circular that telephone prescriptions should only be employed in special circumstances is thus far from fulfilled. The present authors find the the doctors' duty to provide the indications for use should be emphasized on account of control of prescriptions in the pharmacies and ensuring correct medication for the patients concerned. Similarly is should be deliberated whether the requirement of endorsing prescriptions for reissue of B preparations should be maintained and measures for limitation of the use of telephonic prescriptions should be limited with due respect to the advantages and disadvantages of these.

Denmark↗

Prediction of clean mohair, fiber diameter, vegetable matter, and medullated fiber with near-infrared spectroscopy.

Four experiments were conducted in three separate years to test the utility of near-infrared spectroscopy (NIRS) to predict the clean mohair content of Angora goat fleece. Mohair fleece samples were obtained each year from yearling billies at the conclusion of the Angora Goat Performance Test conducted at the Texas A&M University Research Station, Sonora. In Exp. 1 (n = 293) and Exp. 2 (n = 256), fleeces were scanned with a Pacific Scientific (Silver Spring, MD) near-infrared spectrometer fitted with a fiber-optic probe, and calibrations were developed for clean mohair content. In Exp. 3, 59 mohair fleeces collected at the Texas A&M Research Station in San Angelo were sampled four times each. Each sample was scanned with the same spectrometer in reflectance mode fitted with a transport mechanism. This mechanism allowed the instrument to scan a 15-cm2 segment of the fleece sample. Conventional procedures to determine reference values for mohair yield, vegetable matter content, fiber diameter, and percentage of medullated and kemp fibers were conducted. Prediction equations were developed that related NIR spectra to reference values for yield and diameter parameters and were used to predict mohair characteristics for each fleece sample. The predicted and reference values were subjected to a simple analysis of variance to determine variation within and across samples. In Exp. 1, mohair base was related to NIR spectra with R2 = .46 and standard error of calibration (SEC) = 2.84%. In Exp. 2, similar repeatability errors for mohair base could be obtained for both reference- and NIRS-derived values. Fiber diameter and medullated fibers were poorly related to NIR spectra. When samples were scanned using the transport mechanism (Exp. 3), R2 and SEC were .82 and 1.19% for mohair base and .93 and .98 microm for fiber diameter, respectively. The CV for mohair base and diameter were 1.0 and 1.4%, whereas those for predicted mohair base and diameter were 1.4 and 3.4%, respectively. The increased variation within samples for predicted values represents sampling error and lack of fit between NIRS and the laboratory determined values. When the samples from Exp. 1 and 2 were rescanned with the NIRS transport (Exp. 4), R2 and SEC were .79 and 2.03% for mohair base and .52 and 3.49 microm for fiber diameter. The fiber optic probe would facilitate real-time analysis on the shearing floor, but our data indicate that the spectral limitations so far are too severe. A large sample device such as the transport gave excellent results for predicting mohair base and fiber diameter.

Animals↗