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The late Neandertal supraorbital fossils from Vindija Cave, Croatia: a biased sample?

The late Neandertal sample from Vindija (Croatia) has been described as transitional between the earlier Central European Neandertals from Krapina (Croatia) and modern humans. However, the morphological differences indicating this transition may rather be the result of different sex and/or age compositions between the samples. This study tests the hypothesis that the metric differences between the Krapina and Vindija supraorbital samples are due to sampling bias. We focus upon the supraorbital region because past studies have posited this region as particularly indicative of the Vindija sample's transitional nature. Furthermore, the supraorbital region varies significantly with both age and sex. We analyzed four chords and two derived indices of supraorbital torus form as defined by Smith & Ranyard (1980, Am. J. phys. Anthrop.93, pp. 589-610). For each variable, we analyzed relative sample bias of the Krapina and Vindija samples using three sampling methods. In order to test the hypothesis that the Vindija sample contains an over-representation of females and/or young while the Krapina sample is normal or also female/young biased, we determined the probability of drawing a sample of the same size as and with a mean equal to or less than Vindija's from a Krapina-based population. In order to test the hypothesis that the Vindija sample is female/young biased while the Krapina sample is male/old biased, we determined the probability of drawing a sample of the same size as and with a mean equal or less than Vindija's from a generated population whose mean is halfway between Krapina's and Vindija's. Finally, in order to test the hypothesis that the Vindija sample is normal while the Krapina sample contains an over-representation of males and/or old, we determined the probability of drawing a sample of the same size as and with a mean equal to or greater than Krapina's from a Vindija-based population. Unless we assume that the Vindija sample is female/young and the Krapina sample is male/old biased, our results falsify the hypothesis that the metric differences between the Krapina and Vindija samples are due to sample bias.

Animals↗

Behavioral sampling techniques for feedlot cattle.

Continuous observations are an accurate method for behavioral measurements but are difficult to conduct on large numbers of animals because of extensive labor requirements. Thus, we sought to develop methods of behavioral data collection in feedlot cattle production systems that reasonably approximated continuous sampling. Standing, lying, feeding, drinking, and walking behaviors were examined from 224 h of continuous video from 64 heifers. Experiment 1 (n = 24 heifers) compared continuous behavioral sampling techniques (Continuous) with scan sampling using intervals of 1, 5, 10, 15, 30, and 60 min and time sampling (a technique for the periodic recording of behavior) for the first 10 min out of every 60 min. Means for each scan sampling method did not differ in estimated percentage of duration of behaviors (P > 0.05) from continuous sampling, except for scan sampling with a 60-min interval. Scan sampling with a 60-min interval differed from more frequent scan sampling intervals for all behaviors except lying. Scan sampling with short intervals (1 and 5 min) was correlated highly with Continuous for all behaviors. The longer the scan interval, the lower the correlations, especially for behaviors with short duration. Time sampling was not an accurate technique for measuring the sampled behaviors. Focal animal sampling (using continuous sampling of individuals) indicated that one heifer was representative of the entire pen of 10 animals (Continuous) for all maintenance behaviors except drinking. Scan sampling methods (1-, 5-, 10-, and 15-min intervals) were accurate methods of behavioral sampling for feedlot cattle, but scan intervals of 30 or 60 min were less accurate and less precise. Time sampling was not an accurate method because it overestimated standing and underestimated lying behaviors. Experiment 2 (n = 40 heifers) investigated the number of focal animals required to accurately represent continuous behavioral sampling for all animals. Focal animal sampling was accurate for most behaviors using as few as 1 animal out of 10 but was not an accurate method for drinking behavior unless 40% of the animals in the pen were observed. Estimates of sample sizes needed for experimental protocols are provided. Behavioral means, standard deviations, and coefficients of variation are presented along with estimates of required sample sizes. These results validate accurate, precise, and efficient methods for quantifying feedlot cattle behavior.

Animals↗

Bilateral sampling of the internal jugular vein to distinguish between mechanisms of adrenocorticotropic hormone-dependent Cushing syndrome.

BACKGROUND: Bilateral sampling of the petrosal sinuses to distinguish Cushing disease from the ectopic adrenocorticotropic hormone (ACTH) syndrome is accurate but technically demanding and risky. Bilateral sampling of the internal jugular vein is simpler and safer. OBJECTIVE: To compare bilateral internal jugular vein sampling with bilateral inferior petrosal sinus sampling for distinguishing patients with Cushing disease from those with the ectopic ACTH syndrome. SETTING: Tertiary referral hospital. PATIENTS: 20 patients with surgically proven Cushing disease and 1 patient with proven ectopic ACTH syndrome. INTERVENTION: All patients underwent petrosal sinus sampling and jugular vein sampling before and after administration of corticotropin-releasing hormone (CRH) on separate days. MEASUREMENTS: Ratios of central ACTH to peripheral ACTH in petrosal sinus samples and jugular vein samples were calculated before and after administration of CRH. RESULTS: Ratios of central to peripheral ACTH were diagnostic for Cushing disease (> 2 before administration of CRH and > 3 after administration of CRH) in jugular vein samples from 16 of 20 patients with surgically proven Cushing disease (sensitivity, 80% [95% CI, 56% to 96%]). Ten of these 16 patients (63%) had diagnostic results only after CRH was administered. The average ratio of central to peripheral ACTH in jugular vein samples was 2.7 before CRH and 7.7 after CRH. Ratios of central to peripheral ACTH were diagnostic in petrosal sinus samples from 19 of 20 patients with surgically proven Cushing disease (sensitivity, 95% [CI, 75% to 99%]). Samples from all 19 patients had diagnostic ratios before and after administration of CRH. The average ratio of central to peripheral ACTH in petrosal sinus samples was 17.7 before CRH and 90.0 after CRH. In the patient with the ectopic ACTH syndrome, ratios of central to peripheral ACTH were negative in jugular vein samples and petrosal sinus samples before and after CRH (< 2 and < 3, respectively). CONCLUSIONS: Jugular vein sampling correctly identified ACTH-secreting pituitary adenomas in 80% of patients with proven Cushing disease. Administration of CRH was essential for diagnostic results in 63% of the patients. Jugular vein sampling is less invasive than petrosal sinus sampling. Negative results on jugular vein sampling should be confirmed by petrosal sinus sampling.

Adolescent↗

Adaptive sampling in behavioral surveys.

Studies of populations such as drug users encounter difficulties because the members of the populations are rare, hidden, or hard to reach. Conventionally designed large-scale surveys detect relatively few members of the populations so that estimates of population characteristics have high uncertainty. Ethnographic studies, on the other hand, reach suitable numbers of individuals only through the use of link-tracing, chain referral, or snowball sampling procedures that often leave the investigators unable to make inferences from their sample to the hidden population as a whole. In adaptive sampling, the procedure for selecting people or other units to be in the sample depends on variables of interest observed during the survey, so the design adapts to the population as encountered. For example, when self-reported drug use is found among members of the sample, sampling effort may be increased in nearby areas. Types of adaptive sampling designs include ordinary sequential sampling, adaptive allocation in stratified sampling, adaptive cluster sampling, and optimal model-based designs. Graph sampling refers to situations with nodes (for example, people) connected by edges (such as social links or geographic proximity). An initial sample of nodes or edges is selected and edges are subsequently followed to bring other nodes into the sample. Graph sampling designs include network sampling, snowball sampling, link-tracing, chain referral, and adaptive cluster sampling. A graph sampling design is adaptive if the decision to include linked nodes depends on variables of interest observed on nodes already in the sample. Adjustment methods for nonsampling errors such as imperfect detection of drug users in the sample apply to adaptive as well as conventional designs.

Behavior↗

Comparison of three sampling and analytical methods for measuring m-xylene in expired air of exposed humans.

Three breath sampling and analytical methods were tested following exposure of 12 subjects for 4 hr to 75 ppm m-xylene in a controlled environmental chamber. Mixed-expired breath was sampled for m-xylene from all 12 subjects with a new stainless steel device that permits continuous mainstream or sidestream sampling of the solvents present. The m-xylene was sampled from the mainstream using charcoal cloth and from the sidestream using Tenax TA. Alveolar breath also was sampled for m-xylene from 6 of these subjects using bags. The carbon dioxide concentrations of the mixed and alveolar samples, obtained from these 6 subjects, were also determined and used to assess the accuracy of the mixed-expired sampling and analytical procedures. Breath sampling was conducted over the immediate 240-min postexposure period. All m-xylene samples were analyzed using gas chromatography with flame ionization detection. Carbon dioxide concentrations were determined with an infrared analyzer. Nonlinear regression analysis was used to model the desaturation of m-xylene via the breath. Overall, the desaturation of m-xylene from all subjects by all methods was best described using three-compartment pharmacokinetic models. The precision of each sampling and analytical method, estimated from the residual variabilities of the desaturation curves were 0.13 for alveolar sampling, 0.14 for mainstream-mixed sampling (12 subjects), and 0.23 for sidestream-mixed sampling (12 subjects). For all 12 subjects, the breath m-xylene concentrations determined by sidestream-mixed sampling averaged 83% of those determined by mainstream-mixed sampling; this bias was significant. For the 6 subjects from whom both mixed-expired and alveolar breath samples were obtained, the average m-xylene desaturation rates determined by both mainstream-mixed and alveolar sampling were comparable but substantially different from those determined by sidestream-mixed sampling. For these subjects, comparison of the average and individual mixed to alveolar ratios of m-xylene and carbon dioxide showed that mainstream-mixed sampling was accurate and that sidestream-mixed sampling was not.

Air Pollutants, Occupational↗

Evaluation of a sampling method for the measurement of occupational exposures to ethylene.

In an effort to assess the scope of occupational exposures to ethylene, the Olefins Panel of the American Chemistry Council designed and conducted a research project to develop and apply a sampling and analytical method to measure workplace exposure. The method uses packed Carbosieve S-III thermal desorption tubes (Supelco, Bellefonte, Pa.) with a low sample collection flow rate. The tubes were analyzed by thermal desorption gas chromatography. The methodology was validated for both 15-min short-term exposure limit and 8-hr time-weighted average (TWA) samples in the laboratory prior to the field study. The effects of varying sampling flow rate (2, 5, 10, and 25 mL/min) and temperature (25 and 35 degrees C) on sample breakthrough time were assessed at a constant relative humidity of 90%. Breakthrough times decreased linearly with sampling flow rate and temperature. The optimal sampling flow rate and temperature at 90% relative humidity were 2 mL/ min and 25 degrees C. A full-shift TWA sample can be collected using two tubes for up to 4 hours each at a flow rate of 2 mL/min, while a STEL sample can be collected at 25 mL/min flow rate. The evaluation indicated samples can be stored under ambient conditions for a period up to 14 days without significant sample loss. Field measurements were performed at 14 petrochemical facilities within North America. The mean 8-hour TWA ethylene concentration (71 sample pairs) was 2.6 ppm (range: <0.05 to 2100 ppm). Significant ethylene concentrations were observed for only two of the 73 TWA sample pairs. Each of these two samples was obtained from the same facility, and only one tube of the sample pairs showed a high ethylene concentration (3200 ppm and 4200 ppm, respectively) for the 1600 ppm and 2100 ppm TWA sample. The first tube of each of these two sample pairs showed no detectable levels. Further, 69 of 71 sample pairs had TWA concentrations below 13 ppm. The mean of 26 short-term exposure limit samples was 16 ppm (range: <0.05 to 63 ppm), with only one sample above 50 ppm. The results of this study indicate that airborne concentrations of ethylene can be effectively measured using Carbosieve S-III packed thermal desorption tubes under typical workplace conditions.

Adsorption↗

Effect of sampling time and repetition on gingival crevicular fluid and aspartate aminotransferase activity.

Tests based on the composition of gingival crevicular fluid (GCF) for detection of active periodontitis require a better understanding of sampling variables than currently exists. We have studied the effects of sample time and repetition on the presence and activity of aspartate aminotransferase (AST). Two 30-second samples of GCF were harvested within 10-minute intervals from 192 teeth with periodontitis. GCF sample size and AST activity were measured. GCF volume and AST activity of first samples were each approximately 10% greater than for second samples. The differences were significant. AST activity correlated positively with gingival index scores and probing pocket depth. Samples were also harvested from groups of 4 teeth during 5-, 10-, 20- and 30-s periods with 1-min intervals and varying sample order. For these samples, first samples contained the greatest amount of enzyme activity, regardless of sample time. When only first samples were considered, the 5- and 10-s samples showed more total activity than the 20- and 30-s samples, and differences were statistically significant. Flow rate for the 5-s sample was always higher than for all other samples, regardless of its position in the sampling sequence. Our observations are consistent with the existence of a reservoir of AST activity that is, in major part, depleted during the first 5 to 10 s of sampling, and that requires a time period of more than 10 min to reequilibrate. Five- to 10-s samples may distinguish active disease better than 20- or 30-s samples, since most of the activity is taken onto the strip in the first few seconds, and the activity is subsequently partly inactivated or diluted by the uptake of fluid less rich in enzyme activity.

Aspartate Aminotransferases↗

Isolation of Mycobacterium avium subsp paratuberculosis from environmental samples collected from farms before and after destocking sheep with paratuberculosis.

OBJECTIVE: To determine whether Mycobacterium avium subsp paratuberculosis could be isolated from soil-pasture, faecal, water and sediment samples on farms before and after removal of sheep with paratuberculosis. A feasibility study and subsequent field survey. PROCEDURE: First the analytical sensitivity of radiometric culture of the organism from two types of soil was determined relative to faeces. Then soil-pasture, faecal, water and sediment samples were collected for culture from a range of sites from 6 farms with paratuberculosis affected sheep and goats. Similar samples were collected from 20 farms at least 9 months after removal of infected stock. RESULTS: The analytical sensitivity of culture of M a paratuberculosis from soil samples was 2 orders of magnitude less than that from faeces, and environmental samples required longer incubation periods to yield significant growth in radiometric culture (BACTEC) medium. However, the organism was recovered from approximately 20% of 163 soil-pasture, water and sediment samples from 6 properties with clinically-affected animals with paratuberculosis. The positive samples were from a range of topographic sites, including open exposed and dry areas, however, low lying areas tended to have larger numbers of organisms. When the same sites were sampled again about 5 months later, only 1 was culture positive, and none were culture positive > 12 months later. Of 17 water and dam sediment samples collected from farm 6, which had long-standing high prevalence OJD infection, only one water sample and one sediment from the same dam were culture positive. None of the 5 water samples from the other farms were culture positive. Of 96 water samples, 90 sediment samples and 93 soil samples from farms that had been destocked of infected sheep/goats for 9 to 24 months, one sediment sample from a farm in Victoria (destocked for 12 months) and two sediment samples from a farm in New South Wales (10, 19 months) were culture positive. Recontamination from cattle or water could not be excluded as a cause of the positive cultures from the second farm. CONCLUSION: M a paratuberculosis can be detected by radiometric culture in environmental samples from farms grazed by sheep or goats with paratuberculosis. There is a relatively low likelihood of recovery of the organism from water samples from such farms, and at 5 or more months after removing stock with paratuberculosis the likelihood of positive cultures from environmental samples is very low. Although the analytical sensitivity of culture from environmental samples is less than that from faeces, surveys of environmental sites are nevertheless feasible. However, improved culture methods are needed for critical surveys and to study the movement and fate of the organism in the environment.

Animals↗

Comparison of sampling techniques for measuring the antimicrobial susceptibility of enteric Escherichia coli recovered from feedlot cattle.

OBJECTIVE: To evaluate the effectiveness of various sampling techniques for determining antimicrobial resistance patterns in Escherichia coli isolated from feces of feedlot cattle. SAMPLE POPULATION: Fecal samples obtained from 328 beef steers and 6 feedlot pens in which the cattle resided. PROCEDURE: Single fecal samples were collected from the rectum of each steer and from floors of pens in which the cattle resided. Fecal material from each single sample was combined into pools containing 5 and 10 samples. Five isolates of Escherichia coli from each single sample and each pooled sample were tested for susceptibility to 17 antimicrobials. RESULTS: Patterns of antimicrobial resistance for fecal samples obtained from the rectum of cattle did not differ from fecal samples obtained from pen floors. Resistance patterns from pooled samples differed from patterns observed for single fecal samples. Little pen-to-pen variation in resistance prevalence was observed. Clustering of resistance phenotypes within samples was detected. CONCLUSIONS AND CLINICAL RELEVANCE: Studies of antimicrobial resistance in feedlot cattle can rely on fecal samples obtained from pen floors, thus avoiding the cost and effort of obtaining fecal samples from the rectum of cattle. Pooled fecal samples yielded resistance patterns that were consistent with those of single fecal samples when the prevalence of resistance to an antimicrobial was > 2%. Pooling may be a practical altemative when investigating patterns of resistance that are not rare. Apparent clustering of resistance phenotypes within samples argues for examining fewer isolates per fecal sample and more fecal samples per pen.

Animals↗

Differences in total protein concentration, nucleated cell count, and red blood cell count among sequential samples of cerebrospinal fluid from horses.

OBJECTIVE: To examine total protein concentration and cell counts of sequentially collected samples of CSF to determine whether blood contamination decreases in subsequent samples and whether formulas used to correct nucleated cell count and total protein concentration are accurate. DESIGN: Case series. ANIMALS: 22 horses. PROCEDURE: For each horse, 3 or 4 sequential 2-ml samples of CSF were collected from the subarachnoid space in the lumbosacral region into separate syringes, and blood was obtained from the jugular vein. Total protein concentration, nucleated cell count, and RBC counts were determined in all samples. RESULTS: Among 3 sequential samples, total protein concentration and RBC count were significantly lower in samples 2 and 3, compared with sample 1. Nucleated cell count was significantly lower in sample 3, compared with sample 1. Among 4 sequential samples, total protein concentration and RBC count were significantly lower in samples 2, 3, and 4, compared with sample 1. Nucleated cell count was significantly lower in samples 3 and 4, compared with sample 1. For 3 correction formulas, significant differences in corrected values for nucleated cell count and total protein concentration were detected between sample 1 and sample 3 or 4. CONCLUSIONS AND CLINICAL RELEVANCE: Because iatrogenic blood contamination decreases in sequential CSF samples, a minimum of 3 samples should be collected before submitting the final sample for analysis. Formulas to correct nucleated cell count and total protein concentration are inaccurate and should not be used to correct for blood contamination in CSF samples.

Animals↗

Levels of fungi and mycotoxins in samples of grain and grain dust collected on farms in Eastern Poland.

Ten samples of stored wheat grain and 10 samples of settled grain dust released during machine threshing of wheat grain were collected on 10 farms located in Lublin province (eastern Poland). The samples were examined for the concentration of total microfungi, Fusarium species, fusariotoxins (moniliformin, deoxynivalenol, nivalenol), and ochratoxin. Microfungi able to grow on malt agar were present in 30% of grain samples (median for all examined samples = 0, range 0-227.5 x 10(3) cfu/g) and in all samples of grain dust (median = 977.5 x 10(3) cfu/g, range 115.0-16,700.0 x 10(3) cfu/g). Fusarium species (F. avenaceum) were found only in 10% of grain samples (median = 0, range 0-800.0 x 10(3) cfu/g), but in 90% of grain dust samples (median = 1,150 x 10(3) cfu/g, range 5.5-10,060.0 x 10(3) cfu/g). The species F. avenaceum, F. culmorum, F. graminearum, F. poae and F. sporotrichioides were isolated respectively from 50%, 10%, 20%, 40% and 20% of examined grain dust samples. The presence of the mycotoxins produced by Fusarium (moniliformin, deoxynivalenol, and nivalenol) was found altogether in 70% of wheat grain samples (median = 0.1275 microg/g, range 0-1.480 microg/g) and in 90% of grain dust samples (median = 0.350 microg/g, range 0-1.090 microg/g). Moniliformin (MON), deoxynivalenol (DON), and nivalenol (NIV) were each detected in 40% of grain samples, and respectively in 80%, 40%, and 40% of grain dust samples. Ochratoxin A (OTA) was detected in 60% of grain samples and in 60% of grain dust samples (median in both cases was 0.0005 microg/g). The concentrations of F. poae (p<0.05) and of total Fusarium species (p<0.01) in grain samples, and the concentrations of F. culmorum and F. graminearum (p<0.05) in grain dust samples were significantly correlated with the concentration of deoxynivalenol. The concentrations of F. poae (p<0.05) and of total Fusarium species (p<0.01) in grain dust samples were significantly correlated with the concentration of total fusariotoxins. Moreover, the concentration of total Fusarium species in grain dust samples was significantly correlated with the concentration of nivalenol (p<0.05). In conclusion, the majority of samples of wheat grain and grain threshing dust collected on farms in eastern Poland contained notable quantities of fusaria and/or fusariotoxins. This fact poses a potential risk of mycotoxicoses to agricultural workers exposed to grain dust when handling wheat during threshing, unloading, shuffling, and other farm occupations.

Agricultural Workers' Diseases↗

Effects of sample handling on total carbon dioxide concentrations in canine and feline serum and blood.

OBJECTIVE: To determine whether underfilling blood collection tubes leads to in vitro reduction in serum measured total CO2 concentration ([TCO2]m) in canine and feline blood samples sufficient to create the impression of metabolic acidosis (pseudometabolic acidosis) or high anion gap. SAMPLE POPULATION: Blood samples from healthy client-owned animals (16 dogs, 17 cats). PROCEDURE: Venous blood samples were collected in random order for determination of serum [TCO2] and blood gas tensions. Blood gas analysis was performed on iced, capped blood samples. In dogs, serum [TCO2] was measured in 1-, 3-, and 10-ml samples in 10-ml type-B tubes and in a 3-ml sample in 3-ml type-A tubes. In cats, serum [TCO2] was determined in 1-, 2-, and 3-ml samples in 3-ml type-A tubes and in a 3-ml sample in 10-ml type-B tubes. RESULTS: For dogs, serum [TCO2] in full-tube, 10-ml samples was a mean +/- SD, 2.0 +/- 1.1 mmol/L greater than that in 3-ml samples and 3.7 +/- 1.3 mmol/L greater than the value in 1-ml samples; both differences were significant at P < 0.0001. The serum [TCO2] in full 3-ml samples was lower by 0.4 +/- 0.6 mmol/L than the value in full-tube 10-ml samples (P = 0.019). For cats, serum [TCO2] in full-tube, 3-ml samples was 0.5 +/- 0.6 mmol/L greater than that in 2-ml samples (P = 0.004) and was 1.5 +/- 0.8 mmol/L greater than the value in 1-ml samples (P < 0.0001). Serum [TCO2] in 3-ml samples of feline blood in 10-ml tubes was 0.8 +/- 0.8 mmol/L lower than that in samples from full 3-ml tubes (P = 0.0007). In dogs and cats, [TCO2] in fully filled collection tubes was approximately 6 mmol/L higher when calculated from blood gas analysis data than when chemically determined in serum. CONCLUSIONS AND CLINICAL RELEVANCE: Underfilling blood collection tubes results in a false decrease in serum [TCO2], which can contribute in part to descrepancies between blood gas and chemical analyses as estimates of plasma bicarbonate concentration. This, and other in vitro effects of sample handling and collection, may result in a false assessment of metabolic acidosis in dogs and cats.

Acidosis↗

Effect of different sampling techniques on odds ratio estimates using hospital-based cases and controls.

Potential biases introduced by the use of hospital admission records have rarely been discussed in the veterinary literature. Veterinary Medical Teaching Hospital (VMTH) patient records kept at the University of California, Davis (UCD) School of Veterinary Medicine provide a unique opportunity to perform in-depth analyses on the effect of different control selection (sampling) techniques on odds ratio (OR) estimates for disease risk factors in a retrospective case-control study. Horses with Corynebacterium pseudotuberculosis abscesses (134) and the (secondary) study base population (source for controls) were identified, and a 'gold standard' OR for each category of the factors admission type, age, breed and sex was derived. Example data were used to calculate sampling ratios (SRs), defined as the ratio between any sample proportion (of an arbitrary risk factor) and the study base proportion for this risk factor. Sampling ratios different from 1.0 introduced biases into the observed OR estimates, when compared with the 'gold standard' OR. Three randomized samples (simple random, stratified random, systematic sampling), one matched (on date of admission) and three different diagnosis samples ('colic', 'cuts and lacerations', 'fractures') were selected from the study base, and the SRs for all categories of the four factors were derived. The matched and two different disease samples ('colic' and 'fractures') had especially wide ranges of observed SRs (and large errors in the OR estimates), whereas simple random and systematic sampling had comparably narrow ranges (less biased OR estimates). For the three randomized sampling techniques under study, repeated sampling was used to derive SR distributions. The SRs were approximately normally distributed. Analysis of variance and covariance showed that simple random and systematic sampling provided SR distributions with means closest to 1.0 (expected value) and small standard deviations. The OR estimates obtained from records selected by these two sampling techniques therefore were least biased. The findings demonstrate the importance of selecting appropriate sampling techniques in addition to properly defining the study (base) population. Sampling design introduces uncertainty into the OR estimates. The direction of the bias, however, depends on the OR between factor and disease in the source population (the 'gold standard'), and on the direction and magnitude of the SR. When combining the results from single and repeated sampling we conclude that sampling design is most influential on the range of the observed SRs (single samples), on the absolute deviation of the SR from 1.0 (expressed as SR delta Mean) and on the SR standard deviation (SD) (repeated sampling).

Animals↗

Reprocessing unsatisfactory ThinPrep Papanicolaou test specimens increases sample adequacy and detection of significant cervicovaginal lesions.

BACKGROUND: The objective of the current study was to determine the effect of reprocessing bloody ThinPrep (TP) samples using a glacial acetic acid technique on the unsatisfactory rate. METHODS: During a 12-month study period, all TP gynecologic samples received by the Cytology Laboratory from inpatient, outpatient, and community-based clinics at the University of Texas Southwestern Medical Center and the Parkland Health and Hospital System in Dallas, Texas, were enrolled prospectively into the study. The initial TP slides were evaluated for specimen adequacy based on the 2001 Bethesda System. Any TP sample that contained abnormal cells, by definition, was not considered unsatisfactory. The criteria for reprocessing included scant cellularity in a background of abundant blood. Biopsy correlations for all abnormal cytologic diagnoses were established between TP smears and follow-up biopsies. RESULTS: During the course of the 1-year study period, a total volume of 57,296 TP samples were evaluated by the study laboratory. Prior to reprocessing, the laboratory unsatisfactory rate was 8.32% (4767 of 57,296 TP samples). After reprocessing those samples that were compromised by blood (2593 of 4767 TP samples), the unsatisfactory rate was reduced to 5.47% (3134 of 57,296 TP samples), with an overall reduction by 34.25% (2.85 of 8.32 TP samples). Of 2593 samples that were reprocessed, 62.98% (1633 of 2593 TP samples) changed from inadequate to adequate for the purposes of evaluation. Of those 1633 adequate samples, 1509 samples (92.41%) were negative for intraepithelial lesion or malignancy, 72 samples (4.40%) were atypical squamous cells of unknown significance, 5 samples (0.31%) were atypical glandular cells of undetermined significance, 36 samples (2.20%) were low-grade squamous intraepithelial lesions, 10 samples (0.61%) were high-grade squamous intraepithelial lesions, and 1 sample (0.06%) was adenocarcinoma. CONCLUSIONS: The authors conclude that the reprocessing of unsatisfactory ThinPrep cervicovaginal cytology samples decrease the unsatisfactory rate considerably, with an increase in the detection of significant abnormal cervicovaginal lesions.

Female↗

Estimation of Salmonella prevalence on individual-level based upon pooled swab samples from swine carcasses.

Pooling of samples might be an effective means to increase cost-effectiveness in routine surveillance. The present study assessed the effect on the sensitivity of detection of Salmonella when pooling swab samples from swine carcasses compared to individual analyses. A total of 18,984 samples from nine Danish swine abattoirs were collected during 1 year, covering 2017 slaughter days. At each abattoir, swab samples were taken on a daily basis from 10 carcasses randomly selected. From each carcass, an area of 3 cm x 100 cm was swabbed. Five of these samples were analysed individually and the other five were analysed as one pooled sample. Standard culture methods were used. A logistic regression model was built, where the response was whether a sample was Salmonella positive or not. The explanatory factors were abattoir, type of sampling (individual or pooled sample), and season of year 2000 (four quarters). The odds ratio (OR) of the effect of type of sampling in the logistic model accounting for abattoir and season was interpreted as the conversion factor between pooled and individual sample prevalence. The results of the individually analysed samples showed a low prevalence of Salmonella (1.4%). When Salmonella was isolated, mostly only one positive sample was found among the five individually analysed samples per slaughter day. On a few days >1 positive samples' were found (9 out of 2017 days approximately 0.4%). The pooled sample prevalence was 4.1%. Because the individual prevalence was low, the pooled sample prevalence would have been around five times higher than the individual-level prevalence-if there had been no loss of sensitivity. However, we found that due to loss of sensitivity the pooled prevalence was only three times higher (OR = 2.7; CI 2.0-3.7). Therefore, a conversion factor of 3 instead of 5 should be applied to calculate the individual prevalence from a pooled prevalence. This approach has been used in the national surveillance of Danish pork since 2001. The estimated conversion factor and accept of pooling samples do not necessarily apply to a population with a higher prevalence or to other types of samples (e.g. faeces or lymph nodes) or diagnostic procedures.

Abattoirs↗

GLLS for optimally sampled continuous dynamic system modeling: theory and algorithm.

The original generalized linear least squares (GLLS) algorithm was developed for non-uniformly sampled biomedical system parameter estimation using finely sampled instantaneous measurements (D. Feng, S.C. Huang, Z. Wang, D. Ho, An unbiased parametric imaging algorithm for non-uniformly sampled biomedical system parameter estimation, IEEE Trans. Med. Imag. 15 (1996) 512-518). This algorithm is particularly useful for image-wide generation of parametric images with positron emission tomography (PET), as it is computationally efficient and statistically reliable (D. Feng, D. Ho, Chen, K., L.C. Wu, J.K. Wang, R.S. Liu, S.H. Yeh, An evaluation of the algorithms for determining local cerebral metabolic rates of glucose using positron emission tomography dynamic data, IEEE Trans. Med. Imag. 14 (1995) 697-710). However, when dynamic PET image data are sampled according to the optimal image sampling schedule (OISS) to reduce memory and storage space (X. Li, D. Feng, K. Chen, Optimal image sampling schedule: A new effective way to reduce dynamic image storage space and functional image processing time, IEEE Trans. Med. Imag. 15 (1996) 710-718), only a few temporal image frames are recorded (e.g. only four images are recorded for the four parameter fluoro-deoxy-glucose (FDG) model). These image frames are recorded in terms of accumulated radio-activity counts and as a result, the direct application of GLLS is not reliable as instantaneous measurement samples can no longer be approximated by averaging of accumulated measurements over the sampling intervals. In this paper, we extend GLLS to OISS-GLLS which deals with the fewer accumulated measurement samples obtained from OISS dynamic systems. The theory and algorithm of this new technique are formulated and studied extensively. To investigate statistical reliability and computational efficiency of OISS-GLLS, a simulation study using dynamic PET data was performed. OISS-GLLS using 4-measurement samples was compared to the non-linear least squares (NLS) method using 22-measurement samples, GLLS using 22-measurement samples and OISS-NLS using 4-measurement samples. Results demonstrated that OISS-GLLS was able to achieve parameter estimates of equivalent accuracy and reliability in comparison to NLS or GLLS using finely sampled measurements (22-measurement samples), or OISS-NLS using optimally sampled measurements (4-measurement samples). Further more, as fewer measurement samples are used in OISS-GLLS, this algorithm is computationally faster than NLS or GLLS. Therefore, OISS-GLLS is well-suited for image-wide parameter estimation when PET image data are recorded according to the optimal image sampling schedule.

Algorithms↗

Use of EDTA samples for prothrombin time measurement in patients receiving oral anticoagulants.

BACKGROUND AND OBJECTIVES: Oral anticoagulant therapy is commonly called for in health care. Hitherto sampling for prothrombin time (PT) has been measured on blood collected into a coagulation tube and diluted in citrate solution. Blood samples anticoagulated with EDTA are used for hematologic tests and the same sample could also be available for PT. DESIGN AND METHODS: We studied 107 patients on oral anticoagulant therapy. Samples were taken by from both coagulation tubes (citrate) and EDTA tubes. The PT from both samples was measured with an ACL 7000 analyzer and reported in seconds and as an international normalized ratio (INR). The regression equation between citrate and EDTA samples was calculated in both units. We studied the clinical significance of INR results from both sample types and compared the effect of different combined thromboplastin reagents on the correlation equation between citrate and EDTA samples. RESULTS: The regression equation for PT by Owren's PT reagent from citrate (y) and EDTA (x) plasma was y = 1.11 x -0.24 INR, R(2)= 0.99. We observed no clinically significant difference between INR results from citrate and EDTA samples using the regression equation for INR calculation from EDTA samples. ISI depends on sample type (dilution, anticoagulant) and the difference is 0.117, 10%. We calculated ISI for EDTA samples and no clinically significant difference was seen between citrate and EDTA INR results. INTERPRETATION AND CONCLUSIONS: A good correlation was observed between INR results with citrate and EDTA samples from patients receiving oral anticoagulants using Owren's PT reagent with the same citrate calibration. Using the regression equation (INR or sec) for analysis of INR results from EDTA samples is clinically acceptable and offers the possibility of using EDTA samples for PT measurement with citrate calibrators. Reagent international sensitivity index (ISI) values for citrate and EDTA samples differ from each other. ISI determination for EDTA samples requires mathematical calculation of EDTA ISI as in the present study or EDTA-based ISI calibrators. The regression equation for INR from citrate and EDTA samples depended on the reagent used, not only on sample dilution or anticoagulant.

Administration, Oral↗

Detection and identification of significant ANAs in previously determined ANA negative samples.

BACKGROUND: An antinuclear antibody (ANA) substrate transfected with the cDNA to hyperexpress the 60 kD SS-A/Ro antigen (HEp-2000) has been shown to detect anti-SS-A/Ro antibodies missed by standard HEp-2 and other immunoassays. Despite this evidence, many laboratories remain convinced that with experienced technicians, standard HEp-2 is acceptable for ANA detection. AIM: To challenge the ability of HEp-2000 to detect anti-SS-A/Ro antibodies in samples previously determined to be ANA negative using standard HEp-2. METHODS: Three hundred and seventy-one pre-screened "negative" ANA samples were provided by a university hospital laboratory in Germany. These samples were tested on the HEp-2000 substrate at a dilution of 1:40 by indirect immunofluorescence (IIF). Samples that screened positive for a nuclear pattern were titered (range of 1:40-1:640) and all ANA-positive patterns were identified. Samples containing at least one positive ANA pattern at a dilution greater than or equal to 1:160 were further tested. Samples that produced a speckled pattern were tested for antibodies to the extractable nuclear antigens (ENA) and samples that showed homogeneous staining were tested for antibodies to dsDNA, and if negative, were then tested for anti-histone antibodies. RESULTS: Ninety-one patient samples were positive with titers > or =1:160. Speckled patterns were the most common finding (30 samples) followed by speckled/homogeneous mixed patterns (19 samples) and samples demonstrating the SS-A/Ro pattern (16 samples) either alone or in combination with other ANA patterns. The remaining 26 positive samples consisted of various other ANA patterns. The most commonly identified ENAs were SS-A/Ro (14 samples), Scl-70 (11 samples) and SSB (6 samples). No antibodies to dsDNA were identified in 23 positive samples with homogeneous staining patterns, though 17 of these samples tested positive for antibodies to histone. CONCLUSIONS: HEp-2000 detected anti-SS-A/Ro antibodies in 16 (4%) of the "ANA negative" samples. In addition to improved sensitivity for anti-SS-A antibodies, HEp-2000 demonstrated improved sensitivity over standard HEp-2 substrate for other significant ANAs including anti-Scl-70, anti-histone, and anti-SS-B antibodies.

Antibodies, Antinuclear↗