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PubMed2004

Wall-less sample preparation of microm-sized sample spots for femtomole detection limits of proteins from liquid based UV-MALDI matrices.

Previously, we introduced wall-less sample preparation (WaSP), technology that involves the use of an electrodynamic balance (EDB) to prepare microm-sized sample spots for analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In that work we demonstrated the detection of femtomole quantities of a low molecular weight peptide and a hydrophobic ester (both <600 Da). Here we use WaSP to test the hypothesis that the use of small sample spot sizes and an instrument equipped with delayed extraction would increase the analytical utility of liquid sample spots for peptide and protein (>2500 Da) analysis by UV-MALDI-TOF-MS (Sze et al.; J. Am. Soc. Mass Spectrom. 1998, 9, 166-174). To aid the optimization of preparing microm-sized sample spots by WaSP, optical microscopy and mass spectrometry were used to investigate nonvolatile solute concentration effects on droplet fission and sample spot size, modifications of the EDB electric field to control droplet ejection, and the use of multiple droplet deposition to increase sample loading. Also described is a rapid deposition mode of operation for WaSP that allows single droplets generated at 1 Hz to be levitated briefly ( approximately 500 ms) before being ejected autonomously and deposited as a concentrated sample spot with a spatial accuracy of +/-5 microm. To test the sensitivity of the method, one hundred glycerol droplets (270 pL each, 27 nL total) each containing 32 amol lysozyme were deposited on top of each other one-at-a-time to create a single sample spot. Using a mass spectrometer equipped with delayed extraction to analyze this sample spot, we verified the hypothesis of Sze et al. by achieving detection limits three orders of magnitude below that previously observed for the detection of a protein by UV-MALDI-TOF-MS with a chemical-doped liquid matrix sample preparation.

Adrenocorticotropic Hormone↗
PubMed2000

Delayed matching to sample: the effects of sample-set size on human performance.

The discriminative performance of students was assessed on a delayed-matching-to-sample task (DMST) using disks, presented by a computer, as stimuli. The size of the non-matching comparison stimuli was changed, for each participant, until each was 100% correct at 0.05 s delay. Then delays of 0.05, 2, 4, 8 and 16 s were paired with each of one, four, eight, 16, and 32 sample stimuli. Accuracy generally decreased over the one, four and eight samples, did not change consistently over the largest sample-set sizes and decreased as delay increased. Both delay and sample-set size had statistically significant main effects, their interaction was not significant. Fitted exponential functions gave a measure of discrimination at zero delay, a, and a measure of the rate of decrement in performance with increasing delay, b. As number of sample stimuli increased there was no systematic change in b, while a decreased most over one to four samples, decreased less with eight samples, and decreased least from 16 to 32 samples. These results suggest that the effects of varying sample-set size depends on the range of sizes studied, thus, they provide a possible explanation for some previous disparate findings. They also suggest that it might be proactive interference that leads to decreases in accuracy with increasing sample-set size.

Journal Article↗
PubMed1999

The effect of timing of sample collection on the detection of measles-specific IgM in serum and oral fluid samples after primary measles vaccination.

This study compares the timing of the rise and decline of measles-specific IgM in serum samples and in oral fluid samples. Two hundred and eighty 9-month-old infants presenting for routine measles vaccination in Addis Ababa, Ethiopia, were enrolled. Paired serum and oral fluid samples were collected before and 1, 2, 3 or 4 weeks after measles vaccination. Samples were tested by using a modified antibody-capture enzyme immunoassay. For the 321 IgM-negative pre- and post-vaccination serum samples, 317 (99 %) of their corresponding oral fluid samples were IgM-negative. Among the 130 IgM-positive serum samples, 75% of their paired oral fluid samples were IgM-positive, with the percentage rising to 87% after oral fluid samples collected > or =3.5 weeks after vaccination were excluded. Among the post-vaccination serum samples, the percent IgM-positive peaked in week 3 and declined to 79% in week 4. For post-vaccination oral fluid samples, the percent IgM-positive peaked in weeks 2 and 3, and then declined to 43% in week 4. This modified antibody-capture enzyme immunoassay appears to detect vaccine-induced measles-specific IgM in the first 3 weeks after vaccination.

Female↗
PubMed2006

Double-cross hydrostatic pressure sample injection for chip CE: variable sample plug volume and minimum number of electrodes.

A novel sample injection method for chip CE was presented. This injection method uses hydrostatic pressure, generated by emptying the sample waste reservoir, for sample loading and electrokinetic force for dispensing. The injection was performed on a double-cross microchip. One cross, created by the sample and separation channels, is used for formation of a sample plug. Another cross, formed by the sample and controlling channels, is used for plug control. By varying the electric field in the controlling channel, the sample plug volume can be linearly adjusted. Hydrostatic pressure takes advantage of its ease of generation on a microfluidic chip, without any electrode or external pressure pump, thus allowing a sample injection with a minimum number of electrodes. The potential of this injection method was demonstrated by a four-separation-channel chip CE system. In this system, parallel sample separation can be achieved with only two electrodes, which is otherwise impossible with conventional injection methods. Hydrostatic pressure maintains the sample composition during the sample loading, allowing the injection to be free of injection bias.

Electrodes↗
PubMed1997

Probability for detecting hot particles in environmental samples by sample splitting.

The presence of radioactive hot particles in environmental samples (e.g., soil, vegetation, sediments) is frequently detected by observing significant differences in the activities of sub-samples, which are otherwise alike. The probabilities for detecting hot particles in this way were calculated by using Monte Carlo methods as a function of the number of hot particles in the original sample, the number of sub-samples used, the frequency distribution of the activities of the hot particles, and the precision with which the activities of the sub-samples are determined. Assuming, for example, (i) a log-normal distribution of the activities of the hot particles with a relative standard deviation eta > or = 1, and (ii) that a difference of > 30% between the activities of the sub-sample with the largest and that with the smallest activity can be detected, splitting the original sample into three sub-samples will be sufficient to detect the presence of up to five hot particles with a probability of > 95%. If four sub-samples are used, the presence of up to 20 hot particles can be detected with this probability. In general, it will not be effective to increase the precision of the activity measurements of the sub-samples at the expense of the number of sub-samples investigated.

Environmental Pollution↗
PubMed2002

Strategically designed sample composition for fastest screening of polychlorinated biphenyl congeners in water samples.

A new strategy for preparing composite samples of special interest and applicability in environmental screening studies is presented. The use of supersaturated experimental design matrices to conduct the sample composition of water sample specimens in screening studies is demonstrated. In contrast to well known conventional sample composition, this strategic approach provides analytical objects allowing the accurate prediction of analyte concentration levels in the original individual sample specimens while fixing the number of experiments to be carried out down to the very number of sample specimens. This will be of special importance when dealing with analytes that require complicated, labour intensive and expensive analytical processes. To reach this goal, two main conditions must be fulfilled. The first one is the sparsity effect (Pareto principle) which holds for the specimens in the sampling campaign. This means that the number of really anomalous or contaminated specimens, as compared to the total number of specimens to be analysed, is low. In environmental screening studies, frequently this situation can be reasonably assumed. The second condition is to have an effective manner to develop and solve the experimental designs required to build-up the composite samples. The challenging problem of screening PCBs in water samples has been tackled to show the usefulness of this strategic approach by combining chemometrically assisted sample composition and rapid analysis using solid-phase microextraction of the composite samples.

Environmental Monitoring↗
PubMed1992

A study of the performance and comparability of the sampling response to cotton dust of work area and personal sampling techniques.

In order to compare and contrast the sampling response to cotton dust of two forms of dust sampling 85 work areas were identified over a 2-year period for investigation in eight Lancashire spinning mills. Three hundred and five work area dust samples were undertaken and 252 personal dust samples were performed. Operatives who spent a minimum of 80% of their working shift in the area in which work area sampling was also performed were selected for personal sampling. Work area dust exposures have recently shown an upward trend, with highest concentrations occurring in the ring spinning room (median 1.15 mg m-3, range 0.82-2.06). Personal dust samples showed a reduction in dust exposures as cotton processing progressed, from a high in the opening room (median value of 6.24 mg m-3, range 1.0-41.5) to a minimum of 1.02 mg m-3 (range 0.30-0.93) in the winding room. The ratio of measured personal sampling dust exposure to work area sampling exposure was used to compare the relative performance of the two techniques. This ratio was highest in the early processes. There was a 7.8-fold difference in measurement between the two techniques in the opening processes, falling to 4.9 in carding and 4.2 in the other card-room processes. However in ring spinning the ratio was only 1.4, suggesting a degree of comparability in the methods at this stage of processing. The value rose to 2.5 for the last stage (winding). Respiratory disease is known to occur predominantly in the early stages of processing (opening and carding) where high dust concentrations are found using the personal technique. These data support the use of personal sampling for setting exposure limits to cotton dust in preference to the current recommended method using work area sampling techniques, which may significantly underestimate dust exposure in the high risk work areas and is outdated.

Dust↗
PubMed2002

Effect of sampling time on the culturability of airborne fungi and bacteria sampled by filtration.

Air sampling of bioaerosols by filtration may be preferable for many epidemiological studies because the methods can be used to collect personal samples for a full work-shift. There is some concern, however, that the viability of fungal spores and bacterial cells might be compromised by sampling for as long as a full shift. This study was designed to determine the effect of sampling up to 6 h on the viability (measured by culture) of airborne fungi and bacteria at composting facilities. Six side-by-side samples were collected in two locations at each of three composting facilities for 1 h at 2 l/m on polycarbonate filters. Two samples in each set were then capped while clean, HEPA-filtered air was drawn across two others for an additional 2 h and across the last two for an additional 5 h. Filters were washed and the samples were analyzed for culturable bacteria and fungi, and for total bacteria and fungi by microscopic counting. Concentrations ranged from 1.7 x 10(3) to 6.2 x 10(7) c.f.u./m3 of culturable fungi and 1.17 x 10(4) to 1.0 x 10(6) c.f.u./m3 of culturable bacteria. In linear models that included duration of sampling, location, and the interaction of location and sample duration, neither sample duration nor the interaction term were significant predictors of the logs of the concentrations of culturable fungi or bacteria or of the ratio of the logs of the culturable concentrations to total concentrations for fungi or bacteria. This suggests that increased sampling time does not affect the viability of the organisms commonly found in the air at composting facilities.

Air Microbiology↗
PubMed2000

Reconstructing genealogies of serial samples under the assumption of a molecular clock using serial-sample UPGMA.

Reconstruction of evolutionary relationships from noncontemporaneous molecular samples provides a new challenge for phylogenetic reconstruction methods. With recent biotechnological advances there has been an increase in molecular sequencing throughput, and the potential to obtain serial samples of sequences from populations, including rapidly evolving pathogens, is fast being realized. A new method called the serial-sample unweighted pair grouping method with arithmetic means (sUPGMA) is presented that reconstructs a genealogy or phylogeny of sequences sampled serially in time using a matrix of pairwise distances. The resulting tree depicts the terminal lineages of each sample ending at a different level consistent with the sample's temporal order. Since sUPGMA is a variant of UPGMA, it will perform best when sequences have evolved at a constant rate (i.e., according to a molecular clock). On simulated data, this new method performs better than standard cluster analysis under a variety of longitudinal sampling strategies. Serial-sample UPGMA is particularly useful for analysis of longitudinal samples of viruses and bacteria, as well as ancient DNA samples, with the minimal requirement that samples of sequences be ordered in time.

DNA, Viral↗
PubMed1996

Impact of sample selection on APOE epsilon 4 allele frequency: a comparison of two Alzheimer's disease samples.

OBJECTIVE: In a highly selected sample of unrelated Alzheimer's disease (AD) patients, we found that the APOE epsilon 4 allele frequency was higher than previously reported. Differing selection and ascertainment criteria may lead to these differences. To address this possibility, we compared the epsilon 4 allele frequency in two samples of AD patients selected from the same geographical area. SETTING AND PARTICIPANTS: Cases (n = 55) and controls (n = 99) from a research clinic-based sample were compared with subjects (n = 537) from a community-based AD patient sample. The samples consisted of unrelated cases who met NINCDS/ADRDA criteria for probable AD. DESIGN AND MEASUREMENTS: Clinical characteristics and APOE genotype data were obtained from AD cases and controls from both samples. RESULTS: Frequency of APOE epsilon 4 allele in the research cases compared with the community cases (0.45 vs 0.36) was nearly significant. We compared demographic and clinical characteristics that might account for this difference and found that the research cases were younger, had an earlier age of onset, and had more advanced disease than the community cases. After onset age was controlled, there was no overall difference between epsilon 4 allele frequency of the two samples. CONCLUSIONS: We found that the epsilon 4 allele frequency tended to be higher in the research AD sample compared the community-based sample. The two samples differed in several demographic and clinical characteristics. We conclude that research-based samples may lead to enrollment of younger patients with more severe disease who have higher APOE epsilon 4 allele load. This potential selection bias must be considered in the interpretation of studies of APOE allele frequency.

Age of Onset↗
PubMed1993

Verbal self-reports about matching to sample: effects of the number of elements in a compound sample stimulus.

Adults' self-reports about their choices in a delayed matching-to-sample task were studied as a function of the number of elements (one, two, or three) in a compound sample stimulus. Signal-detection analyses were used to examine control of self-reports by the number of sample elements, by the speed and accuracy of choices reported about, and by several events contingent on self-reports. On each matching-to-sample trial, a sample element appeared as one of two comparison stimuli. Choice of the matching element, if made within 500 ms of the onset of the comparison stimuli, produced points worth money or chances in a drawing for money, depending on the subject. After each choice, subjects pressed either a "yes" or "no" button to answer a computer-generated query about whether the choice met the point contingency. The number of sample elements in the matching-to-sample task varied across trials, and events contingent on self-reports varied across experimental conditions. In Experiment 1, the conditions were defined by different combinations of feedback messages and point consequences contingent on self-reports, but self-reports were systematically influenced only by the sample-stimulus manipulation. Self-report errors increased with the number of sample elements. False alarms (inaccurate reports of success) were far more common than misses (inaccurate reports of failure), and false alarms were especially likely after choices that were correct but too slow to meet the point contingency. Sensitivity (A') of self-reports decreases as the number of sample elements increased. In addition, self-reports were more sensitive to choice accuracy than to choice speed. All subjects showed a pronounced bias (B'H) for reporting successful responses, although the bias was reduced as the number of sample elements increased and successful choices became less frequent. Experiment 2 demonstrated that the failure of point contingencies to influence self-reports in the first experiment was not due to a general ineffectiveness of the point consequences. Rates of inaccurate self-reports decreased when they resulted in point losses and increased when they resulted in point gains.

Adolescent↗
PubMed2001

Analysis of factors that may affect the enzymatic profile of gingival crevicular fluid: sampling technique, sequential sampling and mode of data presentation.

To evaluate the possible effect of sampling technique and sequential sampling on gingival crevicular fluid (GCF) volume and myeloperoxidase (MPO) activity, 14 patients presenting at least two symmetrical maxillary sites with mild/moderate periodontitis were selected. Two sites in each individual were sequentially sampled using either the deep-intracrevicular or orifice technique. Spectrophotometrically determined MPO levels were presented either as total MPO activity or MPO concentration. Although the clinical periodontal status of the 20 sampling sites were similar, the deep-intracrevicular technique regularly provided larger GCF volumes. With both techniques, the last samples contained the highest GCF volume. During sequential orifice sampling, GCF volume was relatively more stable. In general, a depletion of MPO activity was observed with sequential sampling performed with either of the techniques. Depletion of MPO did not replenish to baseline levels at the end of the 10-min sequential sampling. Although MPO activity showed a general reduction during sequential orifice sampling with both modes of data presentation, total MPO activity and MPO concentration did not match with the deep-intracrevicular technique. Due to the potential of affecting GCF volume/composition, the selection of sampling technique seems to be a critical methodological decision in GCF-profile studies, primarily during sequential sampling. In GCF-profile studies, mode of data presentation should also be considered.

Adult↗
PubMed2002

The effect of sampling time and sample handling on the detection of Staphylococcus aureus in milk from quarters with subclinical mastitis.

Staphylococcus aureus mastitis is an important cause of economic loss for the dairy industry. Control programs rely on the timely and accurate identification of positive quarters. The effects of sampling time and sample handling were examined in an attempt to improve the accuracy of detection of S. aureus. Premilking and postmilking milk samples were collected from 55 lactating quarters with subclinical S. aureus infection. Each sample was divided into 2 aliquots; one of which was cultured fresh, the other was frozen at -20 degrees C for 14 days before being cultured. Analysis of variance was used to determine the effect of sampling time (premilking vs postmilking) and sample handling (fresh vs frozen) on the detection of S. aureus, as measured by the mean category for colony-forming units per millilitre (cfu/mL). A stratified analysis was required, due to interaction between sampling time and sample handling. Only a fresh postmilking sample was inferior, yielding a lower mean category for cfu/mL (P < 0.05). The ability to detect S. aureus in quarters with subclinical intramammary infection, as measured by the mean category of cfu/mL, was maximized in fresh or frozen premilking samples and in frozen postmilking samples.

Animals↗
PubMed1991

Sampling the difficult-to-sample.

Difficult-to-sample populations are defined as rare populations or populations that are difficult to locate, enumerate or interview. This definition includes subgroups of the United States population that are at increased risk of adverse health effects associated with malnutrition. Examples include persons who are rare (pregnant women), difficult to locate (migrant farm workers), difficult to enumerate (homeless individuals) or difficult to interview (substance abusers). Probability methods to sample rare and elusive populations are reviewed briefly. Methods include disproportionately allocated sampling, multiplicity sampling and the use of multiple frames. The advantages and disadvantages of nonprobability sampling methods are compared using criteria typically applied to assess alternative probability sampling methods. The cost of data collection alone may prohibit consideration of probability sampling methods, but caution is urged before abandoning this statistically sound approach to sample selection. Considerations for sampling the difficult-to-sample are illustrated for one such population, the homeless.

Female↗
PubMed2007

Men who have sex with men in Great Britain: comparison of a self-selected internet sample with a national probability sample.

OBJECTIVES: To compare the characteristics of a self-selected, convenience sample of men who have sex with men (MSM) recruited through the internet with MSM drawn from a national probability survey in Great Britain. METHODS: The internet sample (n = 2065) was recruited through two popular websites for homosexual men in Great Britain in May and June 2003. This sample was compared with MSM (n = 117) from the National Survey of Sexual Attitudes and Lifestyles (Natsal), a probability sample survey of adults resident in Great Britain conducted between May 1999 and February 2001. RESULTS: No significant differences were observed between the samples on a range of sociodemographic and behavioural variables (p>0.05). However, men from the internet sample were younger (p<0.001) and more likely to be students (p = 0.001), but less likely to live in London (p = 0.001) or report good health (p = 0.014). Although both samples were equally likely to report testing for HIV, men from the internet sample were more likely to report a sexually transmitted infection in the past year (16.9% v 4.8%, adjusted odds ratio 4.14, 95% CI 1.76 to 9.74; p = 0.001), anal intercourse (76.9% v 63.3%; p = 0.001) and unprotected anal intercourse in the past 3 months (45% v 36.6%; p = 0.064). CONCLUSIONS: The internet provides a means of recruiting a self-selected, convenience sample of MSM whose social and demographic characteristics are broadly similar to those of MSM drawn from a national probability survey. However, estimates of high-risk sexual behaviour based on internet convenience samples are likely to overestimate levels of sexual risk behaviour in the wider MSM population.

Adolescent↗
PubMed2006

Sample to sample carryover: a source of analytical laboratory error and its relevance to integrated clinical chemistry/immunoassay systems.

BACKGROUND: Integrated systems that combine clinical chemistry and immunoassay analyzers are used routinely. Sample to sample carryover is an inherent risk and can cause erroneously high patient test results for immunoassays. IVD manufacturers and laboratories must be aware of this phenomenon and guard against it. METHODS: We used a sample carryover protocol that directs the clinical chemistry module to process samples with very high immunoassay analyte concentrations followed by samples with very low concentrations for the same analyte. Low concentration samples were then tested by the immunoassay module to determine if the clinical chemistry module caused primary sample tube to primary sample tube carryover of the immunoassay analyte. RESULTS: Sample carryover was assessed on the Abbott ci8200 for HBsAg, AFP, beta-hCG, and PSA. Observed HBsAg carryover met the design specification of <0.1 ppm. Carryover for the other analytes was <0.1 ppm or below the assay limit of detection. CONCLUSIONS: IVD manufacturers must design integrated systems to minimize primary specimen tube carryover and avoid analytical laboratory error that can impact patient safety. Carryover testing is difficult for clinical laboratories to perform in order to verify system performance. Laboratories must consider the potential for specimen carryover and its impact on results whether moving primary sample tubes between separate analyzers or using an integrated system.

Clinical Chemistry Tests↗
PubMed2002

The efficiency of alkaline extraction for the recovery of hexavalent chromium (CrVI) from paint samples and the effect of sample storage on CrVI recovery.

Workplace exposures to CrVI, a human carcinogen, are significant in spraying operations of chromate-containing paints. Accurate determination of CrVI in paint aerosol air samples is important in assessing a worker's exposure to CrVI. National Institute for Occupational Safety and Health method 7604 is widely used for determining CrVI in air samples. It utilizes an alkaline extraction procedure. It was historically validated for paint aerosol samples containing 24.5 to 61.5 microg of CrVI. The literature documented potential airborne CrVI exposures greater than 61.5 microg in recent paint spraying operations. The efficiency of the alkaline method at extracting CrVI from paint samples containing 250 to 3000 microg of CrVI was determined. Paint was prepared, sampled, extracted twice and then digested. Extracts were analyzed for CrVI and digestates of the residual Cr were analyzed for total Cr. Alkaline extraction of paint samples using NIOSH method 7604 resulted in quantitative recoveries for paint samples with CrVI filter loadings from 250 to 3000 microg. A decrease in CrVI extraction efficiency was observed in samples containing > 1000 microg of CrVI. A second extraction improved the recovery of CrVI in these samples. Refrigerating paint aerosol samples for up to 2 weeks did not affect their CrVI content.

Carcinogens↗
PubMed2002

Industrial hygiene sampling for airborne TDI in six flexible slabstock foam manufacturing facilities in the United States: a comparison of the short-term and long-term sampling data.

A majority of all the flexible polyurethane foams are produced by the continuous slabstock process using toluene diisocyanate (TDI) and polyether polyol. The commercially available TDI product is predominantly an 80:20 mixture of the 2,4-TDI and 2,6-TDI isomers. TDI can be used in many applications, but the market is dominated by the flexible slabstock foam industry. Even though there has been a considerable amount of industrial hygiene sampling conducted for airborne TDI in this industry, the focus of the sampling has generally included long-term (time-weighted average) monitoring with a limited amount of sampling for short-term (peak) episodes. By conducting only long-term sampling, which averages the exposure over some extended time period, these short-term episodes (i.e., tasks or activities) of peak exposure were often not characterized. To investigate whether long-term sampling was adequately predictive of short-term excursions, simultaneous (side-by-side) short-term and long-term breathing zone samples were collected on 31 flexible slabstock foam line employees at six manufacturing facilities throughout the United States. Four hundred three short-term samples averaging 15 minutes in length, and 78 long-term samples averaging 124 minutes in length were collected for four operator categories. From these 78 long-term samples, 31 eight-hour TWAs were projected (assuming no further exposure to airborne TDI for the remainder of the work shift) for comparison to the short-term samples. Based on the data collected, compliance with the current ACGIH TLV TWA of 5 ppb would also ensure compliance with the OSHA PEL-C of 20 ppb in a majority of the cases associated with the flexible slabstock foam industry provided that (1) the TLV TWA accounted for the 2,6-TDI isomer in addition to the 2,4-TDI isomer (at this time, exposure limits are published for only the 2,4-TDI isomer), and (2) the TLV TWA was compared to the partial shift TWA measured over the operational time of the workday rather than as an average over an eight-hour workday. Further, exceedences of the 20 ppb PEL-C did occur (e.g., when the foam line tunnel was entered, where there was an "upset" condition, or where there was close contact with hot [fresh cut] buns), but can be protected against with (1) improved work practices, (2) engineering controls, and (3) respiratory protection.

Air Pollutants, Occupational↗