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PubMed1983

Recovery of immune complexes from serum-based samples is dependent on the concentration of reactants: heat-inactivated and unheated samples compared by nephelometric assay.

We examined heat-treated and untreated samples for immune complexes (IC) with a nephelometric type Clq binding assay. Curves of absorbance vs. concentration derived from aggregated gamma-globulin in serum samples showed curvilinear behavior, while curves obtained from aqueous samples were highly linear. Curves from heat-treated sera bent upward, while those from unheated sera tended to level off, causing the curves to cross. Heated sera showed greater recovery of IC in the high range and unheated sera greater recovery of IC in the low. Recovery from serum-based samples varied as the concentrations of both Clq and IC in the reaction mixture changed when aqueous curves were used as calibrators. This behavior in serum-based samples seems to be caused by endogenous binders of IC, and may be an important reason for inter-laboratory variation in results from similar samples.

Antigen-Antibody Complex↗
PubMed1994

Field-amplified sample stacking in micellar electrokinetic chromatography for on-column sample concentration of neutral molecules.

On-column concentration of neutral molecules was achieved for the first time in micellar electrokinetic chromatography by means of field-amplified sample stacking. The stacking process was accomplished by dissolving the neutral analytes in a low-concentration micellar solution that was still above the critical micelle concentration. The lower total ionic strength in the sample buffer compared to the electrophoresis buffer allowed the negatively charged micelles to migrate rapidly into the boundary between the sample and the running buffer where they slow down. This field-amplified sample stacking was achieved by using normal or reversed electrode polarity and produced a 75-85-fold increase in sensitivity for 1,2,4,7- and 1,2,4,8-tetrachlorodibenzo-p-dioxins. The peak area counts obtained from the sample stacking process were proportional to the sample volume injected, and the stacking efficiency was dependent on the micellar concentration. The best stacking efficiency was obtained when the micelle concentration was slightly higher than the critical micelle concentration. When the injection volume was relatively small, the normal-polarity stacking procedure produced a higher stacking efficiency. However, when the injection volume was large, reversed polarity produced a higher stacking efficiency because the non-uniform distribution of the electrical field strength had been eliminated.

Chromatography↗
PubMed1991

The use of national sample surveys for nutritional surveillance: lessons from Malawi's National Sample Survey of Agriculture.

National sample surveys, containing measurements of the weights and heights of children along with other socioeconomic modules, represent one of the important sources of information for nutritional surveillance. The potential uses of such information are political sensitization, targeting by geographic area, targeting by socioeconomic group, and identifying the most promising intervention options according to the presumed causes of malnutrition. The latter two applications depend upon the ability to detect stable associations between nutritional status and socioeconomic factors. This paper examines the extent to which these planning applications are affected by variation in the ecology of malnutrition across different segments of society, using Malawi's National Sample Survey of Agriculture (NSSA). The NSSA is nationally representative of Malawi's smallholder sector and contains information on anthropometrics of underfives and various socioeconomic characteristics of their households. This analysis is based upon 3000 households containing at least one underfive, with one child per household being selected for analysis. Height-for-age Z-scores (HAZ) are calculated using WHO standards, and analyzed using analysis of variance (ANOVA). Variation in nutritional ecology is investigated by examining statistical interactions among socioeconomic variables as they relate to HAZ and by comparing ANOVA models constructed within several sample strata. These strata are defined according to geographic region, size of cultivated area, child's sex, child's age and, for a subsample, ethnic/religious identification. The results demonstrate that significant two-way interactions exist between region, cultivated area, household labor availability and age of the child. For the total sample HAZ declines with increasing cultivated area among young children (less than 24 months) but improves with increasing cultivated area among older children (greater than or equal to 24 months). The overall trend for young children is region-specific, however, such that no generalizations can be made concerning the effects of region and cultivated area without taking the other factor and child's age into account. Similarly, cultivated area interacts with household labor availability among young children, such that more labor is associated with lower HAZ on plots less than 0.7 ha, but more labor is associated with higher HAZ on plots above 1.5 ha. The existence of variation in nutritional ecology is further borne out by the observed variation in multivariable ANOVA models constructed within various sample strata. It is concluded that significant variation in nutritional ecology does exist between various segments of society. This limits the usefulness of national sample surveys for selecting intervention options and for targeting interventions according to socioeconomic characteristics.

Age Factors↗
PubMed2004

New trends in sample preparation: on-line microextraction in packed syringe (MEPS) for LC and GC applications Part III: Determination and validation of local anaesthetics in human plasma samples using a cation-exchange sorbent, and MEPS-LC-MS-MS.

The need for on-line sample preparation for high-throughput applications in bioanalysis has increased during the past decade. In this paper a robust and on-line sample preparation technique, micro extraction in packed syringe (MEPS) has been developed and validated. The method is a miniaturized, fully automated, solid-phase extraction (SPE) technique that can be connected on-line to GC or LC without any modification of the chromatographs. The performance of MEPS as sample preparation method is illustrated by the determination of local anaesthetics in human plasma samples on-line with high performance liquid chromatography (HPLC) and tandem mass spectrometry. The sampling sorbent was 1mg silica based benzenesulphonic acid cation exchanger that was inserted in a 250 microl syringe. Ropicavine and two of its metabolites (PPX and 3-OH-ropivacine), lidocaine and bupivacine were used as model substances. The accuracy values of quality control samples (QC) were between 95% and 109%, and precision (relative standard deviation, R.S.D.) had a maximum deviation of 9% for the analytes.

Anesthetics↗
PubMed1998

Improved method for the simultaneous determination of proguanil and its metabolites by high-performance liquid chromatography and solid-phase extraction of 100-microl capillary blood samples dried on sampling paper.

An improved method is presented for the determination of proguanil, cycloguanil and 4-chlorophenylbiguanide in 100-microl capillary blood samples applied to sampling paper. This method also utilises a solid-phase extraction technique and high-performance liquid chromatography. Different kinds of sampling paper, such as ion-exchange and cellulose sampling paper were tested. The best elution recovery (70-80%) was obtained after treatment of cellulose sampling paper with a quaternary ammonium compound. The limit of determination was 50 nmol/l for cycloguanil and 4-chlorophenylbiguanide and 125 nmol/l for proguanil using 100 microl capillary blood. The stability of the analytes and elution performance from sampling paper was validated at different temperature and storage time. Venous blood and capillary blood concentrations of proguanil and metabolites were found to be similar.

Antimalarials↗
PubMed2003

Effect of sample preparation, length of time, and sample size on quantification of total lipids from bovine liver.

The objective was to evaluate the effect of sample preparation (pulverization under liquid nitrogen, homogenization, or sonication), time length of sonication (0-60 s), shaking in chloroform/methanol solvent (0, 2, 4, or 12 h), incubation in chloroform (0 or 12 h), and drying of extracted lipids at 50 degrees C (2, 4, 6, or 24 h), and sample size (50-250 mg) on quantification of total lipids from bovine liver. Pulverization under liquid nitrogen yielded the lowest recovery. Sonication was least time-consuming for sample preparation. Precise estimates and the greatest recovery were obtained with 30 s of sonication, at least 2 h of shaking in chloroform/methanol solvent, 12 h of incubation in chloroform, and at least 6 h of drying. Sample sizes of at least 150 mg gave precise estimates. The results demonstrate that sample preparation, time length of different steps of the extraction procedure, and sample size affect quantification of total lipid from bovine liver.

Animals↗
PubMed2000

Determination of lead in fish samples by slurry sampling electrothermal atomic absorption spectrometry.

Ultrasonic slurry sampling electrothermal atomic absorption spectrometry (USS-ETAAS) was been applied to the determination of lead in several fish samples. The influences of instrument operating conditions and slurry preparation on the signal were examined. Palladium and ammonium nitrate were used as the modifier to improve the signal. Since the sensitivity to lead in various fish slurries and aqueous solutions was different, the standard additions method was used for the determination of lead in these fish samples. The method was applied to the determination of lead in dogfish muscle reference material (DORM-2) and a swordfish muscle sample purchased from the local market. The analysis results agreed with the reference value. The accuracy was better than 6%. The precision between sample replicates was better than 16% with the USS-ETAAS method. The detection limit of lead estimated from standard additions curve was about 0.053-0.058 microgram g-1 in different samples.

Animals↗
PubMed2002

DNA profiling: a valuable tool for quality control of sample logistics including occurrences of suspected sample confusion in a blood donation centre.

BACKGROUND AND OBJECTIVES: A molecular method for analysing whole-blood samples should be established for quality control of plasma sample logistics. MATERIALS AND METHODS: DNA profiles of retention samples (plasma) were compared to profiles of recent donations (whole blood). DNA extraction, amplification and detection were performed using the Qiagen DNA Blood Mini kit, the AmpFFISTR Profiler Plus Kit and capillary electrophoresis, respectively. RESULTS: Matched pairs of full profiles were obtained for all samples investigated, therefore no deviation from the standardized procedures was detected. CONCLUSIONS: Modified extraction and amplification protocols enabled DNA profiling to be used for the quality control of plasma samples. Hence, DNA profiling can be used in the blood bank as a safe and easy method for quality control of sample logistics.

Blood Banks↗
PubMed2004

Army ants algorithm for rare event sampling of delocalized nonadiabatic transitions by trajectory surface hopping and the estimation of sampling errors by the bootstrap method.

The most widely used algorithm for Monte Carlo sampling of electronic transitions in trajectory surface hopping (TSH) calculations is the so-called anteater algorithm, which is inefficient for sampling low-probability nonadiabatic events. We present a new sampling scheme (called the army ants algorithm) for carrying out TSH calculations that is applicable to systems with any strength of coupling. The army ants algorithm is a form of rare event sampling whose efficiency is controlled by an input parameter. By choosing a suitable value of the input parameter the army ants algorithm can be reduced to the anteater algorithm (which is efficient for strongly coupled cases), and by optimizing the parameter the army ants algorithm may be efficiently applied to systems with low-probability events. To demonstrate the efficiency of the army ants algorithm, we performed atom-diatom scattering calculations on a model system involving weakly coupled electronic states. Fully converged quantum mechanical calculations were performed, and the probabilities for nonadiabatic reaction and nonreactive deexcitation (quenching) were found to be on the order of 10(-8). For such low-probability events the anteater sampling scheme requires a large number of trajectories ( approximately 10(10)) to obtain good statistics and converged semiclassical results. In contrast by using the new army ants algorithm converged results were obtained by running 10(5) trajectories. Furthermore, the results were found to be in excellent agreement with the quantum mechanical results. Sampling errors were estimated using the bootstrap method, which is validated for use with the army ants algorithm.

Journal Article↗
PubMed1999

Synovial fluid sampling from the temporomandibular joint: sample quality criteria and levels of interleukin-1 beta and serotonin.

The aims of this study were to compare two sets of quality criteria (SQC A and B) with respect to synovial fluid (SF) sampling and to present temporomandibular joint (TMJ) SF levels of IL-1 beta and 5-HT. The study comprised 310 TMJ SF samples from 12 healthy individuals (HI) and 59 patients with TMJ inflammatory disorders. Ten HI and 37 patients were selected for investigation of TMJ SF levels and samples were obtained by a push-and-pull method with quantification by vitamin B12. The SQC comprised aspirate weight (AW), dilution factor (DF), blood contamination and hemolysis. IL-1 beta and 5-HT levels did not differ between the samples that satisfied SQC A or B. The proportion of samples that satisfied SQC A was higher than for SQC B. Patients with polyarthritides had significantly higher TMJ SF concentrations of 5-HT and IL-1 beta than HL. In conclusion, there is a recovery of TMJ SF of 0.1-0.2 g with the method used and the criteria set with the highest success rate do not differ from the other one with respect to SF levels of IL-1 beta and 5-HT. This set of sample quality criteria comprised no hemolysis, no or only minor blood contamination, AW > 0.5 g and DF < 0.98. The higher SF levels in the diseased TMJ (polyarthritides) compared to the healthy joint with respect to 5-HT and IL-1 beta is of clinical diagnostic relevance and the presence of 5-HT or IL-1 beta in TMJ SF seems to indicate a pathological joint condition probably of an inflammatory nature.

Adolescent↗
PubMed2006

Two methods of self-sampling compared to clinician sampling to detect reproductive tract infections in Gugulethu, South Africa.

OBJECTIVES: To assess the validity, feasibility, and acceptability of 2 methods of self-sampling compared to clinician sampling during a speculum examination. GOAL: To improve screening for reproductive tract infections (RTIs) in resource-poor settings. STUDY DESIGN: In a public clinic in Cape Town, 450 women underwent a speculum examination and were randomized to self-sample with either a tampon or vaginal swabs. All specimens were tested for the same pathogens using the same diagnostic tests. RESULTS: Self-sampling resulted in satisfactory validity for N gonorrhoeae, C trachomatis, bacterial vaginosis, and Candida species (tampons and swabs) and high-risk human papillomavirus (swabs only) when tested with molecular tests or microscopy, but not for T vaginalis by culture. Self-sampling was feasible and acceptable, but some women preferred speculum examinations, which allow the clinician to view the vagina and cervix. CONCLUSIONS: Although self-sampling should not replace speculum examinations in all circumstances, it should be explored further as an RTI screening strategy.

Adolescent↗
PubMed2005

The effect of cleaning and disinfecting the sampling well on the microbial communities of deep subsurface water samples.

Our knowledge of the microbial characteristics of deep subsurface waters is currently very limited, mainly because of the methods used to collect representative microbial samples from such environments. In order to improve this procedure, a protocol designed to remove the unspecific, contaminant biofilm present on the walls of an approximately 800 m deep well is proposed. This procedure included extensive purges of the well, a mechanical cleaning of its wall, and three successive chlorine injections to disinfect the whole line before sampling. Total bacterial counts in water samples decreased from 2.5 x 10(5) to 1.0 x 10(4) per millilitre during the cleaning procedure. Culture experiments showed that the first samples were dominated by sulfate-reducers and heterotrophs, whereas the final sample was dominated by oligotrophic and hydrogenotrophic bacteria. Community structures established on the diversity of the 16S rRNA genes and data analysis revealed that the water sample collected, after a purge without removal of the biofilm, was characterized by numerous phyla which are not representative of the deep subsurface water. On the other hand, several bacterial phyla were only detected after the full cleaning of the well, and were considered as important components of the subsurface ecosystem which would have been missed in the absence of well cleaning.

Bacteria↗
PubMed2003

Sampling patients within physician practices and health plans: multistage cluster samples in health services research.

OBJECTIVE: To better inform study design decisions when sampling patients within health plans and physician practices with multiple analysis goals. STUDY SETTING: Chronic eye care patients within six health plans across the United States. STUDY DESIGN: We developed a simulation-based approach for designing multistage samples. We created a range of candidate designs, evaluated them with respect to multiple sampling goals, investigated their tradeoffs, and identified the design that is the best compromise among all goals. This approach recognizes that most data collection efforts have multiple competing goals. DATA COLLECTION: We constructed a sample frame from all diabetic patients in six health plans with evidence of chronic eye disease (glaucoma and retinopathy). PRINCIPAL FINDINGS: Simulations of different study designs can uncover efficiency gains as well as inform potential tradeoffs among study goals. Simulations enable us to quantify these efficiency gains and to draw tradeoff curves. CONCLUSIONS: When designing a complex multistage sample it is desirable to explore the tradeoffs between competing sampling goals via simulation. Simulations enable us to investigate a larger number of candidate designs and are therefore likely to identify more efficient designs.

Cluster Analysis↗
PubMed1990

Influence of the interval between removal and freezing of muscle samples on muscle fibre size with special reference to sample size and fibre type.

To study the influences of the interval between removal and freezing of a muscle sample on fibre size, rat hindlimb muscles (soleus and extensor digitorum longus, EDL) were cut into three or five pieces and frozen at different intervals, cross-sectioned, stained and analysed morphometrically. As in a previous study (Larsson & Skogsberg 1988), a pronounced interval-dependent change in soleus muscle fibre size was observed, the fibres in soleus muscle samples frozen immediately (within 15 s) being significantly (P less than 0.01-0.001) larger than those samples frozen 4 or 10 min after removal. However, the interval-dependent change was only noted in small muscle samples, i.e. those where the maximum muscle fibre length was 6.5 mm, while samples in which the fibres were approximately twice this length showed no change. In the slow-twitch soleus, type I fibres predominate, and type II fibres were often too few in one or more cross-sections in each animal to allow reliable measurements of fibre size. The fast-twitch EDL was therefore studied, and the results indicated an interval-dependent change in fibre size which was differentiated according to enzyme-histochemical type, i.e. there was a significant change in type I and IIA fibres but not in type IIB. It is concluded that the interval between removal and freezing of small muscle samples, as in percutaneous muscle biopsies in man, is of vital importance for fibre size, and that this interval should be at least 2-4 min.

Animals↗
PubMed2004

Effects of urinary tract inflammation and sample blood contamination on urine albumin and total protein concentrations in canine urine samples.

BACKGROUND: Urinary tract inflammation and hemorrhage are believed to be common causes of proteinuria in dogs based on results of studies that measured total urine protein concentration. A method to quantify urine albumin (UAlb) concentration in dogs recently has become available; however, the effect of inflammation on albuminuria is unknown. OBJECTIVES: The goals of this study were to determine the effects of urinary tract inflammation, as indicated by pyuria and sample blood contamination, on UAlb concentration and on urine protein:creatinine (UPC) ratio in dogs. METHODS: Urine samples were obtained from dogs with pyuria that were presented to a veterinary teaching hospital or were part of a laboratory colony. To mimic the effects of hematuria, canine whole blood was added to a microscopically normal canine urine sample that had baseline albumin and total protein concentrations below the limits of detection. UAlb concentration was measured using a canine albumin-specific competitive ELISA. UPC ratio was determined using routine methods. RESULTS: Of 70 samples with pyuria, 67% had negligible UAlb concentrations and 81% had normal UPC ratios. UAlb concentration but not UPC ratio was significantly higher (P < 0.05) in samples with concurrent hematuria or bacteriuria. When whole blood was added to normal urine, UAlb concentration did not exceed 1 mg/dL until the sample became visibly pink; the UPC did not exceed 0.4 at any dilution. CONCLUSIONS: Many dogs with pyuria do not have albuminuria or proteinuria; however, albuminuria may be more likely in dogs with pyuria and concurrent hematuria or bacteriuria. Hematuria may not cause an increase in UAlb concentration until it becomes macroscopic and even then may not increase the UPC ratio.

Albuminuria↗
PubMed1992

Detection of Borrelia burgdorferi DNA in urine samples and cerebrospinal fluid samples from patients with early and late Lyme neuroborreliosis by polymerase chain reaction.

A polymerase chain reaction (PCR) was developed for use in the identification of a 248-bp fragment of the Borrelia burgdorferi flagellin gene in urine and cerebrospinal fluid (CSF) from patients with Lyme neuroborreliosis. The specificities of the PCR products were confirmed by DNA-DNA hybridization with an internal probe. The assay had a detection limit of 10 in vitro-cultivated B. burgdorferi. The PCR assay seemed to be species wide as well as species specific, since DNA from all 21 B. burgdorferi isolates from humans tested but not from Borrelia hermsii or Treponema pallidum could be amplified. We tested 10 consecutively diagnosed patients with untreated neuroborreliosis. There was lymphocytic pleocytosis and intrathecal B. burgdorferi-specific antibody synthesis in the CSF of all patients. Urine and CSF samples were investigated by PCR before, during, and up to 8.5 months after therapy. B. burgdorferi DNA was detected in urine samples from nine patients; five patients, including two patients with chronic neuroborreliosis, were PCR positive prior to treatment, whereas urine samples from the remaining four patients obtained 3 to 6 days after the onset of therapy became PCR positive. All urine samples obtained greater than 4 weeks after therapy were negative by PCR. PCR of CSF was less sensitive, and samples from only four patients, including one with chronic neuroborreliosis, were positive. We conclude that urine is a more suitable sample source than CSF for use in B. burgdorferi DNA detection by PCR. Normalization of inflammatory CSF changes and the negative PCR results during follow-up even in patients with chronic neuroborreliosis do not point to a persistent infection. The future role of PCR as a diagnostic tool for Lyme neuroborreliosis is still uncertain.

Adolescent↗
PubMed1998

Rapid detection of Mycobacterium paratuberculosis in clinical samples from ruminants and in spiked environmental samples by modified BACTEC 12B radiometric culture and direct confirmation by IS900 PCR.

The suitability of a radiometric culture medium consisting of BACTEC 12B with PANTA PLUS, mycobactin J, and egg yolk was evaluated for detection of Mycobacterium paratuberculosis in feces, mesenteric lymph nodes, and intestinal walls from cattle, sheep, and goats. In addition, a simple method that would enable the rapid identification of Mycobacterium paratuberculosis by IS900 PCR in the primary cultures was sought so that subculture to secondary egg-free radiometric medium could be avoided. An ethanol extraction followed by differential centrifugation was used to separate M. paratuberculosis from PCR inhibitors in the primary culture. PCR was then undertaken with the pellet, after boiling to lyse the mycobacteria; if this test was negative, the DNA in the lysate was purified with guanidine thiocyanate and silica. Cultures of feces, ilea, and mesenteric lymph nodes from cattle, sheep, and goats known to have or suspected of having Johne's disease yielded positive PCR results 1 to 7 weeks after inoculation. Similar results were obtained with soil and pasture samples that had been spiked with M. paratuberculosis. The results suggested that radiometric culture was more sensitive than histopathology in detecting M. paratuberculosis infection in sheep and goats and more sensitive than culture on Herrold's egg yolk medium for the detection of the infection in cattle. Of 259 individual PCR tests with samples from cultures with growth indices of > or = 10,237 (91.5%) were positive, with only 28 (11.8%) requiring both ethanol and silica preparation to yield a positive result. Of the 22 negative PCR results for samples from cultures with growth indices of > or = 10, 18 were for samples from cultures that had only just developed evidence of growth. PCR-positive cultures tended to remain PCR positive over successive weeks. Flexibility in the timing of the sampling for PCR is thus possible, facilitating batch processing of samples in large-scale disease control programs for ruminants.

Animals↗
PubMed1983

Sample-specific ratio effects in matching to sample.

In a symbolic matching-to-sample task, pigeons were trained using sample-specific, fixed-ratio "observing responses." Subsequently, in a mixed condition, each sample was presented equally often with each ratio requirement, i.e., the ratios were no longer correlated with the samples. In a second experiment, pigeons were trained initially in the mixed condition and subsequently shifted to the sample-specific condition in which the required ratios were correlated with the samples. Results of both experiments suggested joint control of choices by ratio value and by the exteroceptive stimuli. The discriminative properties of the ratios appeared to outweigh absolute ratio-size effects.

Journal Article↗