[Platelet count in EDTA-dependent pseudothrombocytopenia--application of MgSO4 as an anticoagulant].
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Monitoring diabetes is critical for our understanding of the etiology and natural history of disease and for public health actions. However, traditional methods for monitoring are either too expensive (e.g., IDDM registries, NIDDM-OGTT prevalence surveys) or too inaccurate (routinely collected data or passive surveillance) for broad accurate, national programs for monitoring the incidence and prevalence of disease. We suggest that one technology called capture-recapture would considerably increase our ability to "count" diabetes, both nationally and globally. Implementation of this approach could lead to accurate inter- and intracountry data on rates of disease. Moreover, such tracking of diabetes could serve as the model for the monitoring of all disease in the 21st century and beyond.
A series of basic animal studies using a new subatmospheric pressure technique (The V.A.C.) to expedite wound healing are presented. The technique entails placing an open-cell foam into the wound, sealing the site with an adhesive drape, and applying subatmospheric pressure (125 mmHg below ambient) that is transmitted to the wound in a controlled manner. Utilizing a pig model, four studies were undertaken to determine the effect of subatmospheric pressure on laser Doppler-measured blood flow in the wound and adjacent tissue (N = 5), rate of granulation tissue formation (N = 10), clearance of bacteria from infected wounds (N = 5), and measurement of nutrient flow by random-pattern flap survival (N = 5). Blood flow levels increased fourfold when 125 mmHg subatmospheric pressure was applied. Significantly increased rates of granulation tissue formation (p < or = 0.05) occurred with both continuous (63.3 +/- 26.1%) and intermittent (103% +/- 35.3%) application. Tissue bacterial counts significantly decreased (p < or = 0.05) after 4 days of application. Random-pattern flap survival significantly increased (p < or = 0.05) by 21% compared to controls. We determined that the application of controlled subatmospheric pressure creates an environment that promotes would healing.
We initially developed a new flow cytometric (FCM) reference method for the enumeration and staging of nucleated red blood cells (NRBC) in 1997 [Wang et al., 1998 (XIth International Symposium on Technological Innovations in Laboratory Haematology, Banff, Canada, 1998); Tsuji et al., 1999 (Cytometry, 1999)]. The method used CD45 antibody and propidium iodide staining to separate NRBCs from other cells. Accuracy and precision were enhanced because larger numbers of cells were counted than was possible with the manual method. We also developed a method for automated NRBC counting on a haematology analyser, the XE-2100 (Wang, 1988). NRBC were separated from other cells using a special lysing buffer and a fluorescent dye. The XE-2100 was found to detect peripheral and cord blood NRBC accurately and precisely when compared with cell morphology or FCM control methods. The FCM NRBC staging method was established through the identification of different NRBC populations following the novel staining and lysing method. To evaluate the method further, we sorted samples containing NRBCs using a FACSort and investigated NRBC staging on the Sysmex XE-2100 based on the cell sorting results. Data were analysed using special software (ida). First, we used the data in various parameter combinations. We then established gates to classify the NRBC populations. Finally, we analysed blood specimens from patients with different types of diseases to explore possible clinical applications.
We show here an automated (50 samples/h) assay for serum IgG4 having a throughput time of 40 min per sample and a sensitivity of 10 micrograms/ml. The assay procedure is based on the inhibition by sample of the agglutination reaction between monoclonal anti-IgG4 antibodies and latex particles to which IgG4 myeloma protein has been coupled. Assay reliability was ascertained by testing for linearity, analytical recovery (96.4%), interassay precision (less than or equal to 8%), specificity and correlation between the results obtained with monoclonal and polyclonal anti-IgG4 antibodies (n = 84; rs = 0.97). Application of the assay to sera from various groups of patients indicated significantly (p less than 0.00005) higher geometrical means (Gx) in patients suffering from atopy (n = 87; Gx = 617 micrograms/ml), atopic dermatitis (n = 28; Gx = 1,043 micrograms/ml), filariasis with Onchocerca volvulus (n = 48; Gx = 1,681 micrograms/ml) and Brugia malayi (n = 20; Gx = 1,078 micrograms/ml) as compared to nonatopic subjects (n = 103; Gx = 302 micrograms/ml) and randomized paired maternal/cord sera (n = 41; Gx = 276 and 296 micrograms/ml, respectively). IgG4 in the paired maternal/cord sera correlated (r = 0.98; p less than 0.00005). There was no significant influence of age or sex on the IgG4 levels either among the nonatopics or the atopics even though low IgG4 (less than or equal to 30 micrograms/ml) was more common among women. The results suggest that IgG4 and IgE responses are somehow closely related in atopic and parasite-infested patients at the physiological, pathogenic or genetic level.
We describe techniques for the application of two methods, robust to the presence of "outliers", to the hierarchical analysis of variance of bacterial count data from collaborative trials. The techniques are tested against both artificially-generated data with known distributional parameters and actual trial results containing outliers. The relative merits of the robust methods are discussed in comparison with conventional ANOVA techniques.
For the first time, a Live/Dead (L/D) Bacterial Viability Kit (BacLight ) protocol was adapted to marine sediments and applied to deep-sea sediment samples to assess the viability (based on membrane integrity) of benthic bacterial communities. Following a transect of nine stations in the Fram Strait (Arctic Ocean), we observed a decrease of both bacterial viability and abundance with increasing water (1250-5600 m) and sediment depth (0-5 cm). Percentage of viable (and thus potentially active) cells ranged between 20-60% within the first and 10-40% within the fifth centimetre of sediment throughout the transect, esterase activity estimations (FDA) similarly varied from highest (13.3+/-5.4 nmol cm(-3) h(-1)) to lowest values below detection limit down the sediment column. Allowing for different bottom depths and vertical sediment sections, bacterial viability was significantly correlated with FDA estimations (p<0.001), indicating that viability assessed by BacLight staining is a good indicator for bacterial activity in deep-sea sediments. Comparisons between total L/D and DAPI counts not only indicated a complete bacterial cell coverage, but a better ability of BacLight staining to detect cells under low activity conditions. Time course experiments confirmed the need of a rapid method for viability measurements of deep-sea sediment bacteria, since changes in pressure and temperature conditions caused a decrease in bacterial viability of up to 50% within the first 48 h after sample retrieval. The Bacterial Viability Kit proved to be easy to handle and to provide rapid and reliable information. It's application to deep-sea samples in absence of pressure-retaining gears is very promising, as short staining exposure time is assumed to lessen profound adverse effects on bacterial metabolism due to decompression.
This article is dedicated to the application of voxel phantoms in whole-body counting calibration. The first study was performed to validate this approach using IGOR, a physical phantom dedicated to fission and activation product (FAP) measurement, and a graphical user interface, developed at the IRSN internal dose assessment laboratory, called OEDIPE (French acronym for the tool for personalised internal dose assessment) associated with the Monte Carlo code MCNP. The method was validated by comparing the results of real measurements and simulations using voxel phantoms obtained from CT scan images of IGOR. To take this application further, two studies were carried out and are presented in this article. First, a comparison was made between the IGOR voxel based phantom (IGOVOX) and a voxel human body (Zubal Phantom) to confirm whether IGOR could be considered as a realistic representation of a human. Second, the errors made when considering sources homogeneously distributed in the body were assessed against real contamination by taking into account the biokinetic behaviour of the radioactive material for two modes of exposure: the ingestion of 137Cs in soluble form and the inhalation of insoluble 60Co several days after acute incorporation.
OBJECTIVE: To evaluate the utility of capture-recapture methods using multiple, routinely collected, computerized data sources to estimate the numbers and prevalence of diabetes. Methods employed for regional and national monitoring of diabetes have been too inaccurate or too expensive. RESEARCH DESIGN AND METHODS: A survey was undertaken that used four sources of ascertainment to identify prevalent cases of known diabetes in community of Northern Italy: diabetic clinic and family physicians, hospital discharges, prescriptions, and reagent strips and insulin syringes. Capture-recapture methods were employed to estimate the number of missing cases and to adjust for undercount to accurately estimate the number of people who had diabetes. RESULTS: We identified 2,069 unique prevalent cases of known diabetes with the intensive case-finding procedure. The diabetic clinic and family physicians data source identified the largest number of cases. The evaluation of the two sample capture-recapture estimates showed that they were all biased downward because of dependencies between sources. Log-linear modeling was employed to take into account the dependence among all data sources and the heterogeneity of diabetic patients. This method estimated that 2,586 cases existed, resulting in an ascertainment-adjusted prevalence of 2.77% (95% confidence interval, 2.44-3.10). Thus, despite the active case identification, approximately 20% could not be identified. However, the number of cases and rates could easily be adjusted using capture-recapture. CONCLUSIONS: The study shows that a two-sample capture-recapture estimate could be very biased if the investigator is not assured that the sources are independent. However, if at least three data sources are employed, log-linear models allow estimation of the number and prevalence rate adjusted for the degree of undercount (in spite of both the dependence of data sources and the heterogeneity of the diabetic population). The critical factor, however, is that the application of multiple sources with capture-recapture methods could be applied across broad geographical areas and across time to have cost-effective monitoring of diabetes at local and national level.
Particle counting immunoassay (PACIA) is shown to be a suitable nonradioisotopic method for the titration of anti-phospholipase A2 antibodies in the sera from honeybee venom-desensitized patients. The sensitivity of the method is equal to the RAST-IgG with a correlation coefficient between the two methods of r = 0.94. Within-day and between-day precisions were 2.5 and 9.2%, respectively. The main advantages of PACIA are: (i) the use of easily prepared non-radioactive reagents having a shelf-life of at least 1 year at -20 degrees C: (ii) partial automation; (iii) calibration with antisera of animal origin, and (iv) the possibility of allergen standardization of honeybee venom extracts.
Investigations into the efficacy of parenteral ivermectin (Pandex) administration for strongylidosis control in donkeys were carried out. The preparation was applied subcutaneously at a dose of 0.2 mg/kg (1 ml/50 kg body weight). One day prior to the treatment and 14 days post-treatment, individual coprological samples were obtained for faecal nematode egg counts and larval culture. The study was performed on 263 donkeys originating from different regions of Bulgaria. Prior to the treatment and 20 days after that, blood samples were obtained from 64 previously infected animals for monitoring of changes in eosinophil leukocyte counts. The subcutaneous application of ivermectin had an efficacy of 96% in terms of reduction of faecal egg counts. In 92.2% of infected donkeys, a complete reduction of faecal eggs count occurred (0 eggs per gram of faeces epg), whereas in the remaining 7.8% of the infected donkeys, the egg counts were reduced by 72%. The reduction in faecal egg counts did not result in changes in eosinophil counts. The results obtained as well as the lack of local changes after the subcutaneous application of ivermectin in donkeys allow us to recommend its use for control of strongyles in donkeys.
A new image analysing system, designed for microphotometric measurement and pattern recognition has been applied in the discrimination of cells from the various phases of the mitotic cycle. The data acquisition procedure is controlled by a programmable electronic unit and involves the combination of the shifting of the microscope moving stages and the scanning of the successive fields by a mechanical device. The data processing is achieved by a computer. The preliminary results we obtained have shown that such a system allows the automatic recognition and counting of the M, G1, S and G2 cells as also the G0 resting cells. The most useful parameters of the cell proliferation kinetics are thus obtained from a single specimen of a cell population.
Egg counts from a simple composite faecal counting procedure using equal amounts of sample from ten sheep were compared statistically against the arithmetic means of the same ten samples prepared by a conventional egg counting method. Forty separate data sets were analysed in an untransformed bivariate plot and after natural logarithmic transformation. A sign test analysis indicated a high degree of similarity between the two data sets. A confidence interval for the composite count (n = 10) was calculated to give a result between five eggs more and 15 eggs less than the arithmetic mean count of the ten samples. When multiple faecal samples are to be examined, the composite method has significant advantages in time saving and increased throughput whilst still providing an accurate result. This technique has been used to monitor gastrointestinal helminthosis and for faecal egg count reduction testing to assess anthelmintic efficacy.
This paper identifies factors that influence the demand for a bass fishing trip taken in the southeastern United States using a hurdle negative binomial count data model. The probability of fishing for a bass is estimated in the first stage and the fishing trip frequency is estimated in the second stage for individuals reporting bass fishing trips in the Southeast. The applied approach allows the decomposition of the effects of factors responsible for the decision to take a trip and the trip number. Calculated partial and total elasticities indicate a highly inelastic demand for the number of fishing trips as trip costs increase. However, the demand can be expected to increase if anglers experience a success measured by the number of caught fish or their size. Benefit estimates based on alternative estimation methods differ substantially, suggesting the need for testing each modeling approach applied in empirical studies.
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AIMS: To develop a miniaturized analytical system for counting of bacteria. METHODS AND RESULTS: Escherichia coli cells were used throughout the experiments. The system consists of a microfluidic chamber, a fluorescence microscope with a charge-coupled device (CCD) camera and syringe pumps. The chamber was made of a silicone rubber (30 x 30 mm and 4 mm high). The E. coli cells were flowed from a micro-nozzle fabricated in the chamber and detected with the CCD camera. The individual cells were indicated as signal peaks on a computer. The cell counts showed a good correlation compared with that of a conventional plate counting method, and results of the simultaneous detection of live and dead cells were also presented. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The system having a small disposable nozzle has the advantages for low cost and safe medical or environmental analysis, when compared with a conventional flow cytometer. This is the first step of the development of a one-chip microbe analyzer.
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It was demonstrated that there is a relation of cell concentration with the parasite reading in an ELISA reader. It was determined that absorbance showed significant values in wavelengths of visible range and the possible minimum wavelength (450 nm)was chosen to guarantee the maximum sensitivity. Likewise, it was confirmed that absorbance significantly increased (p < 0.001) after the plate was filled up, it stabilized in the 40m-2h time interval when applying 100, 150, 200 or 300 ml per well of the various cell concentrations, it was proved that the optimum volume was 150 ml, a r2 = 0,9986 was reached, and the coefficient of correction(p < 0,001) and the gradient(p < 0,001)were highly significant. In the 5 x 10(4) to 1.5 x 10(7) cell/mL interval, there was an average variation coefficient of 1.75% (0.25-3.17%). Under these conditions, the quantification limit was 5,14 x 10(4) cell/mL. Finally, it was shown that the correlation of counting in Neubauer chamber and optical density was significant.