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PubMed1990

Dependence on exercise intensity of changes in electrolyte secretion from the skin sampled by a simple method.

Secreta from the palm and forearm was sampled for 1-min periods by a new technique, using a glass cylinder. Subjects exercised for 10-min periods at successive intensities of 40%, 50% and 65% VO2max with a leg ergometer operated in the supine position. Changes in the concentrations (values) of Na+, K+ and Cl- in their secreta during exercise were investigated. Significant positive correlations were found between the values of any two electrolytes in samples from the palm or the forearm, but the correlations between values for any one of the three electrolytes from the two sites were not significant. Values for concentrations of the electrolytes were significantly higher in samples from the palm than in those from the forearm at rest, 10 min after the beginning of exercise and at the end of exercise. No significant correlation was found between values for electrolytes in samples from the palm and the exercise intensity, but values for Na+ in samples from the forearm increased stepwise with increase in exercise intensity, and similar tendencies were observed for values of K+ and Cl-. The values for the three electrolytes in samples from the forearm, but not the palm, were significantly correlated with values for blood lactate, the percentage of VO2max and the heart rate. These results suggest that the present technique is suitable for successive samplings of secreta from the forearm, and that values for the electrolytes in samples are useful indices of exercise intensity.

Chlorides↗
PubMed1981

Optimal sampling times for pharmacokinetic experiments.

A sequential estimation procedure is presented which uses optimal sampling times to estimate the parameters of a model from data obtained from a group of subjects. This optimal sampling sequential estimation procedure utilizes parameter estimates from previous subjects in the group to determine the optimal sampling times for the next subject. Parameter estimates obtained from the optimal sampling procedure are compared to those obtained from a conventional sampling scheme by using Monte Carlo simulations which include noise terms for both assay error and intersubject variability. The results of these numerical experiments, for the two examples considered here, show that the parameter estimates obtained from data collected at optimal sampling times have significantly less variability than those generated using the conventional sampling procedure. We conclude that optimal sampling and preexperiment simulation may be useful tools for designing informative pharmacokinetic experiments.

Animals↗
PubMed1986

[Cadmium contents in cereal samples in the past--a comparison with the present].

Samples of wheat, rye and barley from previous harvests, some of them dating back even to the past century and placed at the authors' disposal by a number of seed growers and governmental research institutions, were examined for their cadmium content by AAS or ICP-AES. Although no statistically valid data could be obtained owing to the low number of samples, it should nevertheless be stressed that the cadmium content found in cereal samples from harvests of the past were within the range of present-day wheat samples. Thus, the variation in wheat samples from former harvests was 20-100 micrograms/kg which is coincident with an average cadmium content of 50-60 micrograms/kg measured in 2000 what samples from the harvests of the last 10 years as determined in a special survey by the Federal Ministry of Food, Agriculture and Forestry. Comparison of the results for cereals samples from harvests of the past and the samples of the survey conducted by the Ministry does not indicate a trend towards a continuously rising cadmium contents of bread cereals.

Cadmium↗
PubMed1992

A comparison of methods for sampling the deer tick, Ixodes dammini, in a Lyme disease endemic area.

The purpose of this study was to compare the trapping and examining of mice, drag sampling, and CO2-baited traps for their ability to detect the presence and abundance of immature deer ticks, Ixodes dammini, in a Lyme disease endemic area in southern New York State. Eight study sites were sampled 14 times between 28 May and 31 August by setting 49 live-traps, four CO2-baited traps, and drag sampling 500 m2. A total of 1540 nymphs and 3079 larvae was collected during the study. Drag sampling collected the most nymphs (705), while more larvae were recovered from CO2-baited traps (1105). Comparisons among the methods showed a significant difference in the numbers of both larval and nymphal ticks collected (P less than 0.01). There was a positive correlation between the numbers of nymphs collected by drag sampling and CO2-baited tick traps (rs = 0.83, P less than 0.05), and between the numbers of larvae collected by drag sampling and mouse trapping (rs = 0.75, P less than 0.05). These results suggest that drag sampling would be the single most reliable method for quantitatively sampling immature I. dammini populations in a Lyme disease endemic area.

Analysis of Variance↗
PubMed1997

Comparison of a novel microaerobic system with three other gas-generating systems for the recovery of Campylobacter species from human faecal samples.

Three commercial gas-generating systems--CampyGen (Oxoid, UK), Oxoid BR56 (Oxoid, UK), and CampyPak Plus (Becton Dickinson, USA)--and the evacuation replacement technique were compared for the recovery of Campylobacter spp. from 500 human faecal samples collected from patients with gastroenteritis. Four hundred fifty faecal samples were tested upon receipt in the laboratory. Fifty faecal samples that had been previously found to be positive for Campylobacter spp. were tested retrospectively; these had been stored at 4 degrees C for more than 48 h. A total of 41 (9.1%) of the fresh faecal samples and 41 of 50 (82%) of the stored faecal samples were positive for thermophilic campylobacters. The CampyGen, the Oxoid BR56, the CampyPak Plus, and the evacuation replacement system detected Campylobacter spp. in 40 (97.6%), 39 (95.1%), 41 (100%), and 41 (100%) of the positive fresh faecal samples and in 37 (90.2%), 40 (97.6%), 39 (95.1%), and 40 (97.6%) of the stored samples, respectively. There was no statistical difference in performance of any of the four gas systems used (p = 0.98; chi-square test). Eighty-six percent of the isolates were Campylobacter jejuni and 14% were Campylobacter coli. Biotyping and phage typing of the isolates demonstrated that they were of a diverse range of subtypes. This study demonstrates that thermophilic campylobacters can be isolated from human diarrhoeal faecal samples using any of the four microaerobic-atmosphere-generating systems.

Campylobacter↗
PubMed1993

Evaluation of a commercial latex agglutination test for rapid detection of Salmonella in fecal samples.

A latex agglutination test for the detection of salmonella in feces was evaluated in comparison to direct culture and enriched culture using both artificially inoculated samples and clinical samples. In the samples inoculated artificially with different concentrations of salmonella (10(1) to 10(5) per gram) the enriched culture performed better only at the 10(2) level in 0.4 g samples, whereas the latex test performed as well as the enriched culture at all levels in 4 g samples. In the tests using clinical samples, there was no significant difference between results of the latex test performed in 2283 samples and the enriched culture performed in 2072 samples. The sensitivity, specificity and negative and positive predictive values of the latex test were 88.2%, 98%, 97.5% and 63% respectively. The test provided results rapidly but yielded a number of false positive results.

Bacteriological Techniques↗
PubMed1995

Detection of Giardia lamblia cysts in stool samples by immunofluorescence using monoclonal antibody.

The diagnostic potential of indirect immunofluorescence to detect Giardia cysts in stool samples using a cyst-specific anti-Giardia lamblia monoclonal antibody was evaluated in comparison to conventional light microscopy. One hundred fifty specimens from clinically suspected Giardia infections and 50 control samples from microscopically proved Giardia infections were tested. Giardia cysts were found in 15 of 150 (10%) samples tested by light microscopy, whereas immunofluorescence microscopy detected 35 of 150 (23%) positive samples. Forty-six of the 50 reference samples previously shown to contain Giardia cysts were positive. Apparently, the four discrepant samples contained very low numbers of parasites, as none could be detected by conventional microscopy. The results show that Giardia lamblia cysts are detected significantly more frequently using the antibody marker. The doubled number of positive stool specimens and detection of as little as four cysts per sample suggest that microscopical examination of samples can be improved by immunofluorescent staining of Giardia lamblia cysts.

Animals↗
PubMed2004

Quantitative PET cerebral glucose metabolism estimates using a single non-arterialized venous-blood sample.

UNLABELLED: The purpose of this study is to develop a method of quantitating the cerebral metabolic rate of glucose (CMRglc) by positron emission tomography using a population-based heated venous curve and one-point sampling from a non-heated vein, i.e. that can avoid arterial puncture. METHODS: We conducted this study on 17 subjects with a mean age of 61 +/- 9 years. A time-concentration curve as an input function was obtained by sampling 24 blood samples, from the heated left hand vein, one before and the others after intravenous injection of 259 MBq of F-18-fluorodeoxyglucose into the right cubital vein. A non-heated venous sample was also obtained from the right cubital vein. RESULTS: The population-based input function was calculated by averaging time-concentration curves from the first 7 subjects. A single sample obtained from 10 other subjects from 7.5 to 20 minutes and 35 and 40 minutes after injection predicted input function well with an error of less than 4.5%. The radioactivity in the non-heated 40 minutes' sample was 1.7 +/- 2.9% higher than in the heated vein. When we calibrated the population-based curve using the non-heated venous samples at 40 minutes in 10 subjects, the calculated CMRglc values were 1.3 +/- 5.4% lower than the actual values. CONCLUSIONS: Non-heated venous one-point sampling and the population-based curve can decrease the complexity of the procedures and the manpower required, and also make the FDG study less invasive, without a significant increase in measurement error.

Aged↗
PubMed2000

Reappraisal of single-sample and gamma camera methods for determination of the glomerular filtration rate with 99mTc-DTPA.

The aim of this study was to assess the clinical validity of single-sample methods and gamma camera uptake methods with 99mTc-DTPA for the estimation of the glomerular filtration rate (GFR). The study was performed in 50 patients with various degrees of renal function (29 males and 21 females; age 27-90 yrs; serum creatinine level 0.34-6.49 mg/dl). As a reference the "true" GFR (GFRt) was determined from plasma clearance by means of the two compartment model curve fitting 10 plasma samples. The GFRt of more than 30 ml/min in 46 patients was compared to the GFR which was estimated with 7 single-sample methods, two gamma camera uptake methods and 24-hour endogenous creatinine clearance (24hCcr). Close correlation was observed in all single-sample methods. The highest linear correlation was observed in the Christensen and Groth's method rewritten by Watson for a 180-min plasma sample (r = 0.991, see = 5.84 ml/min). The smallest random error was observed in the Groth and Aasted's method for a 180-min plasma (r = 0.989, see = 4.31 ml/min/1.73 m2). Our method was lowest in % absolute difference analysis (mean = 4.10%). The gamma camera uptake methods correlated significantly with the GFRt (r = 0.746-0.774), but were less reliable than any of the single-sample methods (see = 15.41 ml/min-19.14 ml/min). The lowest correlation was observed in the 24hCcr (r = 0.698, see = 50.76 ml/min/1.73 m2). The single-sample method was more accurate than the gamma camera method, and the gamma camera method was more accurate than 24hCcr. The single-sample method should be recommended for the accurate determination of the GFR with 99mTc-DTPA in a patient with mild to moderate renal dysfunction.

Adult↗
PubMed2004

Continuous ultrasound-assisted extraction coupled to a flow injection-flame atomic absorption spectrometric system for calcium determination in seafood samples.

Calcium was extracted on-line from solid seafood samples by a simple and rapid continuous ultrasound-assisted extraction system. This system is connected to a flow injection manifold, which allows the on-line flame atomic absorption spectrometric determination of calcium. This method enables the analysis of solid samples avoiding time-consuming traditional sample preparation methods and their inherent errors. The on-line manifold for calcium determination is the simplest possible, because a volume of 250 microL of acid extract is injected into an ultrapure water carrier stream. The acid extract was diluted on-line with lanthanum, which also acts as masking agent in order to avoid chemical interferences. The continuous monitoring of the calcium signal was accomplished by flame atomic absorption spectrometry. A Plackett-Burman experimental design was used for the optimisation of the continuous leaching procedure. The method allowed a total sampling frequency of 40 samples per hour, with a relative standard deviation for the complete procedure of 0.9% (for a sample containing 3414.35 mg/kg calcium (dry mass)). The limit of detection was found to be 44.4 mg/kg (dry mass) for 5 mg of sample. The analytical procedure was applied to real seafood samples.

Calcium↗
PubMed2004

Slurry sampling of sediments and coals for the determination of Sn by HG-GF AAS with retention in the graphite tube treated with Th or W as permanent modifiers.

A method for the determination of Sn in slurry samples of sediment and coal by hydride generation graphite furnace electrothermal atomic absorption spectrometry (HG-GF AAS) is proposed. The slurries were prepared by mixing the ground sample (particle size < or =50 microm) with 2.0 mol L(-1) HCl for the sediment samples or with 2.0 mol L(-1) HCl+1.0% v/v HF in a saturated boric acid medium for the coal samples. The slurry was placed in an ultrasonic bath for 30 min, before and after standing for 24 h, with occasional manual stirring. The graphite tube was treated with 0.5 mg of Th or W as a permanent modifier. Sn determination was carried out by electrothermal atomic absorption spectrometry at the optimized retention temperatures of 450 and 300 degrees C for Th and W treatment, respectively. With this coupling, kinetic interference in the formation of the hydrides is avoided, and excellent detection limits can be obtained by using peak height. For the chemical vapor generation device, an optimized volume of 2 mL of sample slurry and an optimized NaBH4 concentration of 5% m/v were employed. The vapor produced was transported and retained on the graphite tube surface, which was further heated for Sn atomization. The accuracy of the method was verified by analyzing five certified sediments and three coals. By using the external calibration against aqueous standard solutions, the results obtained were in agreement with the certified values only for the sediment samples. For the coal samples, an addition calibration curve, obtained for one certified coal, was necessary to achieve accurate results. The obtained limits of detection were 0.03 microg g(-1) for sediment and 0.09 microg g(-1) for coal with Th as permanent modifier. The relative standard deviations were lower than 15%, demonstrating an adequate precision for slurry analysis. Sediment and coal samples from Santa Catarina, Brazil, were also analyzed.

Journal Article↗
PubMed2006

Headspace solid-phase microextraction gas chromatography tandem mass spectrometry for the determination of brominated flame retardants in environmental solid samples.

A headspace solid-phase microextraction gas chromatography coupled with tandem mass spectrometry (HSSPME-GC-MS-MS) methodology for determination of brominated flame retardants in sediment and soil samples is presented. To the best of our knowledge, this is the first time that SPME has been applied to analyze polybrominated biphenyls (PBBs) and polybrominated diphenyl ethers (PBDEs) in environmental solid samples. Analyses were performed using 0.5-g solid samples moisturized with 2 mL water, employing a polydimethylsiloxane (PDMS) fiber coating, exposed to the headspace at 100 degrees C for 60 min. Several types of environmental solid samples were included in this study and the extraction efficiency was related to the organic matter content of the sample. Calibration was performed using real samples, and the method showed good linearity over a wide concentration range, precision, and afforded quantitative recoveries. The obtained detection limits were in the sub-ng g(-1) for all the target analytes in both samples. The proposed procedure was applied to several marine and river sediments and soils, some of which were found to contain PBDEs at concentrations in the ng g(-1) level; BDE-47, BDE-100, and BDE-99 were the major congeners detected. The proposed method constitutes a rapid and low-cost alternative for the analysis of the target brominated flame retardants in environmental solid samples, since the clean-up steps, fractionation, and preconcentration of extracts inherent to the classical multi-step solvent extraction procedures are avoided.

Journal Article↗
PubMed2006

Projection of multidimensional GC data into alternative dimensions-exploiting sample dimensionality and structured retention patterns.

Comprehensive multidimensional gas chromatography (GCxGC) is a powerful separation technique. One of the features of this technique is that it offers separations with more apparent structure than that offered by conventional one-dimensional GC (1-D GC). While some previous studies have alluded to this structure, and used structured retention patterns for some simple classifications, the topic of structured retention in GCxGC has not been studied in any great detail. Using the separation of fatty acid methyl esters (FAME) on both nonpolar/polar and polar/nonpolar column sets, the interaction between the separation dimensions and the sample dimensions is explored here. The GCxGC separation of a series of compounds is presented as a projection of the sample from sample space, a p-dimensional space with dimensions defined by the dimensionality of the sample, into separation space: for GCxGC, a two-dimensional plane passing through the sample space in an orientation defined by the separation conditions. Using this conceptual model and some a priori knowledge of the sample, it is shown how the image of the sample in the separation space can be used to construct an image of the sample in alternate dimensions, such as second dimension retention factor ((2)k) vs. chain length in the case of FAME. These projections into alternate dimensions should facilitate the interpretation of the complex patterns found within the GCxGC chromatogram for the identification and classification of compounds.

Journal Article↗
PubMed2001

Development of an ICP-HRIDMS method for accurate determination of traces of silicon in biological and clinical samples.

An inductively coupled plasma-high resolution isotope dilution mass spectrometric (ICP-HRIDMS) method in combination with a microwave-assisted decomposition technique has been developed for the determination of traces of silicon in biological and clinical samples. A 30Si-enriched spike solution was used for the isotope dilution step. Decomposition of the samples was achieved by use either of HNO3 or a mixture of HNO3 and HF. By application of both methods of digestion to the same sample it was possible to differentiate between a poorly soluble silicate fraction and an HNO3-soluble silicon species. Traces of silicon were determined in different reference materials, which are not certified for this element, and in other biological and clinical samples. A concentration range of 1-600 microgram g(-1) was covered by the different samples. For homogeneous samples relative standard deviations of 2-4% were obtained. The detection limit was strongly affected by the blank. In this connection purification of water, used in the analytical procedure, was especially critical. The blank contribution of the ICP-MS instrument could be minimized by applying a nebulizer and a spray chamber made of PFA, a sapphire injection tube, and a silicon nitride torch. Under these conditions detection limits of 0.15 microgram g(-1) and 0.2 microgram g(-1) were obtained for the HNO3 and HNO3-HF digestion methods, respectively, when a sample weight of 0.5 g was used. With regard to expected silicon content this enables determination in almost all biological and clinical samples. The ICP-HRIDMS results were compared with those recently obtained in an interlaboratory study. This isotope-dilution method is an potential option for certification of silicon in reference materials, a method for which is still required.

Animals↗
PubMed2001

Neuronal responses in the frontal cortico-basal ganglia system during delayed matching-to-sample task: ensemble recording in freely moving rats.

Electrophysiological recording of single neuron activity has been conducted in rats to investigate the patterns of distributed neuronal responses in the frontal cortico-basal ganglia system that code information during a spatial-delayed matching-to-sample task (DMTSt). Rats were trained to press one of the two retractable levers presented randomly as a sample response. The first valid nose-poke after a delay resulted in the presentation of both levers. Pressing the same lever as the sample lever led to a water reward (match to sample), whereas pressing the lever opposite the sample lever resulted in a time-out (house light turned off). One hundred seventy-one neurons in the medial prefrontal cortex (mPFC), 51 in the dorsal striatum (STR), and 93 in the nucleus accumbens (NAc) were recorded during DMTSt. Different patterns of neuronal responses were observed during different behavioral episodes (sample, delay, and match phases) in all three recording areas. Space-related neuronal responses specific to the side of the lever pressed were more often found in the sample phase than in the match phase in all three areas studied. Neuronal responses specific to either correct or error trials were observed with similar percentages in the mPFC and the NAc, while the incidence of correct/error-coded activity in the STR was lower. Ensemble neuronal activity that coded sample versus match lever presses was observed in three out of five rats in sets of trials with similar speed and trajectory of lever press. The results reveal specific patterns of neural responses in the frontal cortico-basal ganglia system in rats during the DMTSt and suggest the existence of specific neuronal coding for different behavioral events associated with a learned short-term memory process.

Action Potentials↗
PubMed2002

A femoral arteriovenous shunt facilitates arterial whole blood sampling in animals.

In this study we evaluated on-line continuous blood sampling in a femoral arteriovenous (a-v) shunt for use in quantitative tracer studies using gamma-emitting radionuclides in animals. The shunt consisted of 40 cm polyethylene tubing (PE-50) guided through a coincidence probe. Two three-way valves allowed blood pressure measurements and tracer injection. Blood flow in the shunt and the impulse response function (IRF) were assessed using heparinized human blood mixed with fluorine-18 fluorodeoxyglucose (FDG). In vivo experiments were performed in eight male rats (300-350 g) anaesthetized with halothane. In three rats, manual blood sampling was performed in parallel with on-line sampling. In another five animals, the arterial whole blood activity was recorded on-line for 40 min. For the experiments 150-180 MBq FDG was injected over 35 s. Blood flow in the shunt was 23.6, 29.2 and 42.8 ml/h at 100, 120 and 160 mmHg, respectively. The IRF was characterized by minimal dispersion (1-2 s FWHM). Deconvolution of the measured arterial input curves with the IRF changed the measured curve only minimally. Whole blood radioactivity concentration derived from manual and on-line sampling were in excellent agreement. The curves derived from on-line sampling were of high statistical quality. In conclusion, a femoral a-v shunt allows multiple manipulations such as measurement of the arterial whole blood activity, continuous blood pressure monitoring, injection of the tracer and collection of blood samples if necessary. It is not associated with blood loss if the collection of blood samples is not required. It is more convenient to use than manual sampling, the peak of the input curve is never missed and the input curves are of high statistical quality.

Animals↗
PubMed2004

Development of an optimal sampling strategy for clinical pharmacokinetic studies of the novel anthracycline disaccharide analogue MEN-10755.

AIM: MEN-10755 is a novel anthracycline analogue that has shown an improved therapeutic efficacy over doxorubicin in animal models, especially in gynaecological and lung cancers and is currently under clinical development for the treatment of solid tumours. The aim of the project was to develop an optimal sampling strategy for MEN-10755 to provide an efficient basis for future pharmacokinetic/pharmacodynamic investigations. METHODS: Data from 24 patients who participated in a phase I clinical pharmacokinetic study of MEN-10755 administered as a short i.v. infusion were included. Individual pharmacokinetic values were calculated by fitting the plasma concentration data to a two-compartment model using nonlinear least-squared regression (KINFIT, Ed 3.5). Population pharmacokinetic analysis was carried out using (a) the traditional standard two-stage method (STS) based on all data (KINFIT-ALL), (b) the iterative two-stage Bayesian (IT(2)B) population modelling algorithm (KINPOP), and (c) the STS method using KINFIT and using four optimally timed plasma concentrations (KINFIT-OSS4). Determinant (D) optimal sampling strategy (OSS) was used to evaluate the four most information-rich sampling times. The pharmacokinetic parameters V(c) (l), k(el) (h(-1)), k(12) (h(-1)) and k(21) (h(-1)) calculated using KINPOP served as a model for calculation of four D-optimal sampling times. D-optimal sampling data sets were analysed using KINFIT-OSS4 and compared with the population model obtained by the traditional standard two-stage approach for all data sets (KINFIT-ALL). RESULTS: The optimal sampling times were: the end of the infusion, and 1.5 h, 3.8 h and 24 h after the start of the infusion. The four-point D-optimal sampling design determined in this study gave individual parameter estimates close to the basic standard estimates using the full data set. CONCLUSION: Because accurate estimates of pharmacokinetic parameters were achieved, the four-point D-optimal sampling design may be very useful in future studies with MEN-10755.

Algorithms↗
PubMed2005

Identification and determination of the viability of Giardia lamblia cysts and Cryptosporidium parvum and Cryptosporidium hominis oocysts in human fecal and water supply samples by fluorescent in situ hybridization (FISH) and monoclonal antibodies.

In the present study, fluorescent in situ hybridization (FISH) and monoclonal antibodies (MAbs) were evaluated for species-specific detection and viability determination of Giardia lamblia, Cryptosporidium parvum, and Cryptosporidium hominis in human fecal and water supply samples. A total of 50 fecal human samples positive for G. lamblia cysts, 38 positive for C. parvum, and 23 positive for C. hominis were studied. Also, 18 water supply samples positive for Giardia spp. and Cryptosporidium spp. by the United States Environmental Protection Agency (USEPA) Method 1623 were studied by FISH and fluorescein isothiocyanate (FITC)-conjugated MAbs. Eighteen percent of the fecal samples parasitologically positive for G. lamblia presented viable and nonviable cysts, and 5% of those positive for Cryptosporidium spp. presented viable and nonviable oocysts. Of the 18 water supply samples analyzed, 6 (33%) presented Giardia spp. viable and nonviable cysts and 2 (11%) presented viable and nonviable Cryptosporidium spp. oocysts. G. lamblia identification was confirmed by polymerase chain reaction (PCR) and sequencing of the beta-giardin gene in the fecal and water samples found positive by FISH and FITC-conjugated MAbs. C. parvum and Cryptosporidium muris were identified, by PCR and sequencing of the small subunit of ribosomal RNA gene, in seven and one water samples, respectively. Our results confirm that this technique enables simultaneous visualization, species-specific identification, and viability determination of the organisms present in human fecal and water supply samples.

Animals↗

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