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At least 469 records · Page 26Linked to original sources
PubMed2004

Potential of capillary-column-switching liquid chromatography-tandem mass spectrometry for the quantitative trace analysis of small molecules. Application to the on-line screening of drugs in water.

We have investigated the potential of capillary-column-switching liquid chromatography coupled to tandem mass spectrometry (cLC-MS-MS) for the quantitative on-line trace analysis of target compounds in aqueous solutions. The technical design of the nano-scale cLC system developed at our Institute for peptide and protein identification has been tested and evaluated for the direct trace analysis of drugs in water samples. Sulphametoxazole, bezafibrate, metoprolol, carbamazepine and bisoprolol occurring frequently in Dutch waters, were selected as test compounds. Adequate conditions for trapping, elution and MS-MS detection were investigated by employing laboratory made 200 microm i.d. capillary columns packed with 5 microm aqua C18 material. In the final cLC-MS-MS conditions, a 1 cm length trapping column and a 4 cm length analytical column were selected. Under these conditions, the target compounds could be directly determined in water down to a level of around 50 ng/l employing only 25 microl of water sample. Validation was done by recovery experiments in ground-, surface- and drinking-water matrices as well as by the analysis of water samples with incurred residues and previously analyzed with a conventional procedure involving off-line solid-phase extraction and narrow-bore LC with MS-MS detection. The new methodology provided recoveries (50-500 ng/l level) between 50 and 114% with RSDs (n = 3, each level) below 20% for most of the compounds. Despite the somewhat less analytical performance in comparison to the conventional procedure, the on-line approach of the new methodology is very suitable for screening of drugs in aqueous samples.

Chromatography, High Pressure Liquid↗
PubMed2006

A practical approach to optimization and validation of a HPLC assay for analysis of polyribosyl-ribitol phosphate in complex combination vaccines.

The use of multi-factor statistical experimental design methodology minimized the vaccine material and laboratory resources required for optimization and validation of an HPLC assay for quantitation of depolymerized and total PRP. Components of the assay selected for optimization were adjuvant dissolution, ultracentrifuge conditions including ultracentrifuge model, sample diluent, mobile phase and column oven temperature. Previous experience has shown these components of the assay to be most troublesome and therefore required optimization prior to validation. Specificity, linearity, precision, accuracy and ruggedness were confirmed through a validation of the optimized assay. The validation also established the assay to be stability indicating, by showing that changes to the integrity of the PRP-OMPC conjugate could be detected.

Chromatography, High Pressure Liquid↗
PubMed2006

Observation of anomalous diffusion in excised tissue by characterizing the diffusion-time dependence of the MR signal.

This report introduces a novel method to characterize the diffusion-time dependence of the diffusion-weighted magnetic resonance (MR) signal in biological tissues. The approach utilizes the theory of diffusion in disordered media where two parameters, the random walk dimension and the spectral dimension, describe the evolution of the average propagators obtained from q-space MR experiments. These parameters were estimated, using several schemes, on diffusion MR spectroscopy data obtained from human red blood cell ghosts and nervous tissue autopsy samples. The experiments demonstrated that water diffusion in human tissue is anomalous, where the mean-square displacements vary slower than linearly with diffusion time. These observations are consistent with a fractal microstructure for human tissues. Differences observed between healthy human nervous tissue and glioblastoma samples suggest that the proposed methodology may provide a novel, clinically useful form of diffusion MR contrast.

Algorithms↗
PubMed1996

Umbilical cord blood gas analysis at the time of delivery.

AIMS: it is now recommended that cord blood acid-base measurement is performed routinely at time of delivery in the UK as a measure of fetal response to labour. However, there remains some uncertainty about the value of this procedure. In this paper our experience of cord blood analysis is described and the literature is reviewed to: (1) provide an overview of the physiological basis of cord blood acid-base assessment; (2) describe the appropriate methodology and identify issues which have contributed to confusion and undermined the value of cord blood sampling; and (3) address the practical issues of cord blood sampling. CONCLUSIONS: cord blood acid-base measurement has a sound physiological basis. It provides objective information which is a useful adjunct to subjective methods of newborn assessment, enables babies at risk of neonatal morbidity to be identified, can be helpful in litigation cases and is a prerequisite for clinical audit. However, to be of benefit the information must be correct and correctly interpreted.

Bias↗
PubMed2006

Multiresidue method for the determination of quinolone antibiotics in bovine raw milk by capillary electrophoresis-tandem mass spectrometry.

A new analytical method based on capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) is proposed and validated for the identification and simultaneous quantification of eight quinolones for veterinary use in bovine raw milk. The studied quinolones include danofloxacin, sarafloxacin, ciprofloxacin, marbofloxacin, enrofloxacin, difloxacin, oxolinic acid, and flumequine, whose contents are regulated by the EU Council Regulation no. 2377/90 in animal edible tissues. Different parameters (i.e., separation buffer composition and electrospray conditions) were optimized in order to obtain both an adequate CE separation and a high sensitivity, using experimental design methodology to consider the interactions among the studied variables. MS/MS experiments using an ion trap as analyzer operating in the multiple reaction monitoring mode were carried out to achieve the minimum number of identification points according to the 2002/657/EC European Decision. For the quantification in bovine raw milk samples, a two-step solid-phase extraction procedure was developed using Oasis MAX and HLB cartridges without protein precipitation. Satisfactory results were obtained in terms of linearity (r2 between 0.989 and 0.992) and precision (RSD below 18%). The limits of detection and quantification (below 6 and 24 ppb, respectively) were in all cases lower than the maximum residues limits tolerated for these compounds in milk, the recoveries ranging from 81 to 110%, indicating the potential of the CZE-MS/MS for the analysis of regulated quinolone antibiotics in the food quality and safety control areas.

2-Propanol↗
PubMed2006

Accurately quantifying low-abundant targets amid similar sequences by revealing hidden correlations in oligonucleotide microarray data.

Microarrays have enabled the determination of how thousands of genes are expressed to coordinate function within single organisms. Yet applications to natural or engineered communities where different organisms interact to produce complex properties are hampered by theoretical and technological limitations. Here we describe a general method to accurately identify low-abundant targets in systems containing complex mixtures of homologous targets. We combined an analytical predictor of nonspecific probe-target interactions (cross-hybridization) with an optimization algorithm that iteratively deconvolutes true probe-target signal from raw signal affected by spurious contributions (cross-hybridization, noise, background, and unequal specific hybridization response). The method was capable of quantifying, with unprecedented specificity and accuracy, ribosomal RNA (rRNA) sequences in artificial and natural communities. Controlled experiments with spiked rRNA into artificial and natural communities demonstrated the accuracy of identification and quantitative behavior over different concentration ranges. Finally, we illustrated the power of this methodology for accurate detection of low-abundant targets in natural communities. We accurately identified Vibrio taxa in coastal marine samples at their natural concentrations (<0.05% of total bacteria), despite the high potential for cross-hybridization by hundreds of different coexisting rRNAs, suggesting this methodology should be expandable to any microarray platform and system requiring accurate identification of low-abundant targets amid pools of similar sequences.

Oligonucleotide Array Sequence Analysis↗
PubMed1994

A passive ozone sampler based on a reaction with nitrite.

Standard ozone monitoring techniques utilize large, heavy, and expensive instruments that are not easily adapted for personal or microenvironmental monitoring. For large-scale monitoring projects that examine spatial variations of a pollutant and human exposure assessments, passive sampling devices can provide the methodology to meet monitoring and statistical goals. Recently, we developed a coated filter for ozone collection that we used in a commercially available passive sampling device. Successful preliminary results merited further validation tests, which are presented in this report. The passive ozone sampler used in field and laboratory experiments consists of a badge clip supporting a barrel-shaped body that contains two coated glass fiber filters. The principle component of the coating is nitrite ion, which in the presence of ozone is oxidized to nitrate ion on the filter medium (NO2- + O3 produces NO3- + O2). After sample collection, the filters were extracted with ultrapure water and analyzed for nitrate ion by ion chromatography. The results from laboratory and field validation tests indicated excellent agreement between the passive method and standard ozone monitoring techniques. We determined that relative humidity (ranging from 10% to 80%) and temperature (ranging from 0 degrees C to 40 degrees C) at typical ambient ozone levels (40 to 100 parts per billion) do not influence sampler performance. Face velocity and sampler orientation with respect to wind direction were found to affect the sampler's collection rate of ozone. Using a protective cup, which acts as both a wind screen and a rain cover, we were able to obtain a constant collection rate over a wide range of wind speeds.

Environmental Exposure↗
PubMed1998

Consideration of PCR methods for the detection of SV40 in tissue and DNA specimens.

SV40 is capable of infecting humans, although its association with human diseases remains controversial. Recently, a subgenomic SV40 DNA sequence was detected by polymerase chain reaction (PCR) in certain types of human tumour tissue as well as in normal pituitary tissue. However, due to the limited DNA sequence information that was obtained in those experiments, SV40 could not be authenticated, and it was uncertain whether a related or hybrid virus (or endogenous DNA) accounted for the PCR-amplified DNA. To gain more insight into these observations, we are experimenting with PCR primers directed at various sites of the SV40 genome, as well as with various parameters of the PCR assay. In this communication, we describe methodology we currently use in our studies and discuss problems associated with the PCR detection of SV40 in various types of samples.

Bone Neoplasms↗
PubMed2005

A new method for generating poroelastograms in noisy environments.

Poroelastography has been recently introduced as a new elastographic technique that may be used to describe the spatial and temporal behavior of poroelastic materials. The experimental methodology proposed thus far for phantoms and tissues in vitro requires the acquisition of a precompression rf frame, the application of a unit step strain compression to the sample and the acquisition of subsequent post-compression frames from the material. Elastograms and poroelastograms are generated by cross-correlating the sequentially-acquired postcompression frames with the reference precompression frame. The application of poroelastography to tissues in vivo must address the echo decorrelation problems that are encountered due to uncontrolled tissue motion, which may become significant shortly after the acquisition of the precompression frame. In this paper, we investigate the feasibility of performing poroelastography experiments using an alternative experimental scheme. In the proposed experimental methodology, the reference precompression frame is continuously moved while the time interval between the frames that are correlated is kept short. This allows long data acquisition times with simultaneous minimization of the decorrelation due to undesired tissue motion in vivo. We validated this new method using both a step and a ramp compression functions. We performed poroelastographic simulations and experiments in phantoms and in tissues in vivo. The results were compared to those obtained using the traditional acquisition methodology. This study shows that the two methods yield similar results in vitro and suggests that the new method may be more robust to decorrelation noise in applications in vivo.

Arm↗
PubMed2000

Methodological issues in microdialysis sampling for pharmacokinetic studies.

Microdialysis is an in vivo technique that permits monitoring of local concentrations of drugs and metabolites at specific sites in the body. Microdialysis has several characteristics, which makes it an attractive tool for pharmacokinetic research. About a decade ago the microdialysis technique entered the field of pharmacokinetic research, in the brain, and later also in peripheral tissues and blood. Within this period much has been learned on the proper use of this technique. Today, it has outgrown its child diseases and its potentials and limitations have become more or less well defined. As microdialysis is a delicate technique for which experimental factors appear to be critical with respect to the validity of the experimental outcomes, several factors should be considered. These include the probe; the perfusion solution; post-surgery interval in relation to surgical trauma, tissue integrity and repeated experiments; the analysis of microdialysate samples; and the quantification of microdialysate data. Provided that experimental conditions are optimized to give valid and quantitative results, microdialysis can provide numerous data points from a relatively small number of individual animals to determine detailed pharmacokinetic information. An example of one of the added values of this technique compared with other in vivo pharmacokinetic techniques, is that microdialysis reflects free concentrations in tissues and plasma. This gives the opportunity to assess information on drug transport equilibration across membranes such as the blood-brain barrier, which already has provided new insights. With the progress of analytical methodology, especially with respect to low volume/low concentration measurements and simultaneous measurement of multiple compounds, the applications and importance of the microdialysis technique in pharmacokinetic research will continue to increase.

Animals↗
PubMed1991

The influence of depressive symptomatology on alcoholics' locus of control: a methodological note and a correction.

In a previously published experiment from our laboratory, we reported two findings. First, controls showed the expected negative correlation between internal and external LOC dimensions, while alcoholics did not. Second, a difference score between LOC-internal and LOC-external was correlated significantly and positively with alcoholics' abstracting impairment. Data from two new independent samples of alcoholics and controls confirmed the first finding, but not the second; further, a calculation error was discovered in the previously published data that invalidated the second result. However, in all three studies, depressive symptomatology was found to be correlated negatively with the internal-external LOC difference score in both alcoholics and controls. It is suggested that depressive symptomatology be assessed in any study that investigates LOC.

Adult↗
PubMed2005

Romantic relationships for women with anorexia nervosa: an integrative literature review.

OBJECTIVE: To critically appraise literature concerning romantic relationships for women with anorexia nervosa (AN), and to gain perspective on future research directives. METHOD: Key electronic databases, reference lists, and journals were searched. The search was limited to features of romantic relationships and their quality. Nineteen studies were critiqued and abstracted. RESULTS: All 19 studies used a quantitative design; no qualitative studies were retrieved. Methodological strengths of these studies included confounder control and appropriate statistical analyses for design. Weaknesses included a failure to declare study designs, address sources of bias, and justify study samples, as well as a lack of con-founder control. DISCUSSION: The review drew attention to the varied relational experiences and marked relationship dissatisfaction expressed by women with AN. However, an understanding of relationship quality and its dynamic nature remains limited. Future research should focus on how meanings are negotiated in relationships, providing detailed descriptions of the women's experiences, which is best done with qualitative study. Well-conducted longitudinal research should address questions that cannot be answered by current research designs, such as temporality.

Adult↗
PubMed2004

Genotyping with TaqMAMA.

TaqMAMA combines the quantitative strengths of TaqMan with the allele-specific PCR of MAMA. In this article we develop TaqMAMA as a technique for screening human DNA samples for known genetic polymorphisms. In the first set of experiments, plasmids that model all types of genetic polymorphisms were used to understand the relationship between TaqMAMA primer/template mismatches and their strength of allelic discrimination. These data can be used to improve allelic discrimination of other primer extension genotyping methodologies through directed use of nucleotide mismatches. We used the data to derive a guide for TaqMAMA primer design and DNA strand selection for TaqMAMA genotyping assays. The guide was then used to develop assays for 11 known and novel human genetic polymorphisms. Genotypes were assigned quickly and accurately in all cases. TaqMAMA genotyping assays require minimal development time, have a high probability of success, produce reliable data that are straightforward to analyze, and are very cost-competitive.

Alleles↗
PubMed2001

Evaluating the comprehensibility of jury instructions: a method and an example.

Methodological problems in jury simulation research involve issues of sampling, choice of stimulus materials, appropriate unit of analysis, appropriate dependent variable, corroborative data, and problems of role playing. Despite these issues, comprehension of jury instructions may be suitable for examination by jury simulation techniques--if certain of these methodological concerns can be satisfied. In a series of 5 experiments using typical Canadian legal instructions on criminal conspiracy and the coconspirator exception to the hearsay rule, this study attempted to validate a simple and inexpensive technique for testing the incomprehensibility of a given set of jury instructions by requiring participants to apply those legal instructions to a set of facts. The results demonstrate the utility of an application test, and suggest that for assessing the comprehensibility of jury instructions, it may be acceptable to use undergraduate students as participants, to use individual participants without group deliberation, and to employ written stimulus materials.

Adolescent↗
PubMed2006

Uptake rates for breast cancer genetic testing: a systematic review.

PURPOSE: Individuals and families dealing with the possibility of hereditary cancer risk face numerous decisions, including whether to obtain genetic testing. The purpose of this article is to determine what is known about the rate at which people obtain cancer genetic testing. METHODS: Using MEDLINE, CINAHL, and PSYCHINFO plus reviewing reference lists of relevant articles, we identified 40 studies in May 2002 that addressed breast cancer-related decisions, enrolled adult participants, were published in 1990 or more recently, were peer-reviewed primary clinical studies, addressed genetic testing either alone or in combination with genetic counseling, and reported rates at which participants showed interest in and/or underwent cancer genetic testing. Information regarding study design, participants, and genetic testing uptake rates was recorded. Each article was reviewed for methodologic quality using a flexible quality review system applicable to all study types. RESULTS: Of the 40 studies, 25 provided information about hypothetical genetic testing decisions, 14 about real decisions, and 1 about both. Mean hypothetical uptake was 66% (range, 20-96%) and real uptake was 59% (range, 25-96%). Multivariate logistic regression analyses found that decision type (real/hypothetical), personal and family history of breast cancer, and variability in sampling strategy, recruitment setting, and criteria for real and hypothetical uptake were independently associated with uptake. Our systematic review identified additional explanations for uptake variability (investigator influences, small sample sizes, variability in target populations, lack of clearly described sampling strategies, sampling methods open to bias, and variability in reporting associated risk factors). CONCLUSION: In addition to clinical characteristics, research methodologic issues are likely to be major determinants of variability in published breast cancer genetic testing uptake rates. An understanding of these issues will clarify to clinicians why their clinical experience may not be congruent with published rates and help guide future research.

Adult↗
PubMed1986

Mutagenic activity of disinfection by-products.

Data on raw water quality, disinfection treatment practices, and the resulting mutagenic properties of the treated water were compiled from pilot- and full-scale treatment experiments to evaluate that parameter which might produce variability in the results of a mutagenic study. Analysis of the data and comparison of treatment practices indicated that the measured mutagenic activity is strongly related to the characteristics of the organic matter in the raw water, the methodology used to sample and detect mutagens, the scale of the study both in terms of treatment flow and period of study, and the point at which and the conditions under which oxidants are added during treatment. Conclusions regarding disinfection systems in full-scale water treatment plants include the following: When raw water is pretreated and high concentrations of organics are present in the raw water, both ozonation and chlorination increased mutagenic activity. However, no significant difference in mutagenicity was found between the two oxidants. Both in the case of a nitrified groundwater and a clarified surface water, the mutagenic activity of the water after ozonation was related to its mutagenic activity before ozonation. With ozonation, mutagenic activity decreased after granular activated carbon (GAC) filtration. Thus, when GAC filtration follows ozone disinfection, early addition of oxidants may not be deleterious to the finished water quality. When chlorine or chlorine dioxide is added after GAC filtration, chlorine dioxide was found to produce a less mutagenic water than chlorine. Although these conclusions suggest means of controlling mutagenic activity during treatment, it must be stressed that the measurement of mutagenicity is a presumptive index of contamination level.

Carbon↗
PubMed1990

[Event-related desynchronization and the selection of an alpha-frequency band for quantifying cortical pre- and poststimulus activation].

The amount of event-related decrease of increase in alpha-band power is termed "Event-Related Desynchronization" or ERD. In this study, ERD is used as an index of cortical activation, which not only can be applied to monitor poststimulus but prestimulus processes as well. Usually, ERD is analyzed within a fixed range of alpha-frequency, typically within a range of 8-12 Hz. alpha-frequency, however, shows a considerable between subject variation, even in a sample of subjects with comparable age. The purpose of this study, thus is to show as to how the range of alpha-frequency affects the calculation of ERD. Two different methodological approaches were compared. First, ERD was calculated within a fixed range of 8-12 Hz. Second, ERD was calculated within an individual range, which was adjusted to the individually determined alpha-frequency of each subject. A sample of ten right handed subjects participated in an episodic memory experiment, in which target words, shown in a earlier session of the experiment, were to be identified. A time span of 375 ms preceding and following the presentation of a word was defined as the "pre-" and "poststimulus" interval. EEG-signals were recorded unipolarly from 29 leads. The results show a rather localized increase of ERD in the poststimulus interval. In the prestimulus interval on the other hand, the average magnitude of ERD is much lower and the topographical distribution less localized. Most importantly, however, when comparing the two methods of analyzing ERD, those ERD-values calculated within an individually determined alpha-band, are superior in order to differentiate between the pre- and poststimulus interval.

Adult↗
PubMed1988

Case-control studies of the effect of environmental sanitation on diarrhoea morbidity: methodological implications of field studies in Africa and Asia.

The problems and prospects in the use of case-control studies to assess the effects of improvements in environmental sanitation on diarrhoea morbidity are discussed on the basis of two field studies. It is concluded that an adequate design is available for assessing the effects of a single improvement on diarrhoeal disease. The estimates of effect appear to be valid and sufficiently precise. For addressing more complex questions of interactions, sample sizes would have to be increased substantially. The experience with two field studies suggests that there is hope that a simpler protocol may be feasible, in which only limited information is collected, in which few home visits are made, and in which analytical techniques are simple. Until more field studies have been conducted definitive conclusions cannot be reached on the applicability of such a simple, rapid and inexpensive approach.

Child, Preschool↗