SEARCH · PubMed Health

Results for “Absolute quantification”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,189 records · Page 66Linked to original sources

PubMed2000

Direct in situ measurements of labile inorganic and organically bound metal species in synthetic solutions and natural waters using diffusive gradients in thin films.

The emerging technique of DGT (diffusive gradients in thin films) is shown to be capable of performing new speciation measurements in situ in natural waters. In DGT, metals are bound to a resin layer after passing through a well-defined diffusion layer. Cd was measured in solutions containing glycine, EDTA, and fulvic (FA) and humic acids (HA) by atomic absorption spectroscopy (AAS), anodic stripping voltammetry (ASV), and DGT. DGT measured similar labile fractions to ASV, with detailed differences being consistent with a thicker diffusion layer allowing more dissociation of labile complexes and a slower diffusion of FA and HA complexes through the gel. When single measurements are made in complex solutions with DGT, precise quantification is impossible due to uncertainties concerning the distribution of species with different diffusion coefficients. A new procedure was proposed based on the advantage of DGT of being able to control the pore size of the diffusive gel layer. Small (inorganic) species diffuse freely through all gels but larger FA and HA (organic) complexes diffuse less freely in more constrained gels. When measurements were made on known solutions of Cu and FA or HA, it was possible to quantify the inorganic and organic species separately. They agreed well with predictions made using the WHAM speciation code. Multiple DGT units were also deployed in situ in a stream with high dissolved organic carbon (14.6 mg/L). The systematic differences between the devices with different gel compositions enabled determination, for the first time, of the in situ concentrations of both labile inorganic and organic species in natural water. A single DGT device with a constrained gel can be used to quantify inorganic species directly, providing absolute accuracy is not required. This ability of DGT to measure well-defined fractions of metals in situ using a simple device gives it considerable potential as a regulatory tool. A direct speciation measurement may be preferable to modeling approaches which require diverse input data that are difficult to determine.

Benzopyrans↗

PubMed2004

Quantification of a novel group of nitrate-reducing bacteria in the environment by real-time PCR.

Nitrate reduction is performed by phylogenetically diverse bacteria. Analysis of narG (alpha subunit of the membrane bound nitrate reductase) trees constructed using environmental sequences revealed a new cluster that is not related to narG gene from known nitrate-reducing bacteria. In this study, primers targeting this as yet uncultivated nitrate-reducing group were designed and used to develop a real-time SYBR(R) Green PCR assay. The assay was tested with clones from distinct nitrate-reducing groups and applied to various environmental samples. narG copy number was high ranging between 5.08x10(8) and 1.12x10(11) copies per gram of dry weight of environmental sample. Environmental real-time PCR products were cloned and sequenced. Data was used to generate a phylogenetic tree showing that all environmental products belonged to the target group. Moreover, 16S rDNA copy number was quantified in the different environments by real-time PCR using universal primers for Eubacteria. 16S rDNA copy number was similar or slightly higher than that of narG, between 7.12x10(9) and 1.14x10(11) copies per gram of dry weight of environmental sample. Therefore, the yet uncultivated nitrate-reducing group targeted in this study seems to be numerically important in the environment, as revealed by narG high absolute and relative densities across various environments. Further analysis of the density of the nitrate-reducing community as a whole by real-time PCR may provide insights into the correlation between microbial density, diversity and activity.

Bacteria↗

PubMed1995

Short-term evaluation of plaque area measurement reproducibility from computer-assisted sonography.

One reason why quantifying plaque regression is difficult is the poor spatial control of the shooting angle whether in angiography or ultrasonography techniques. A computer-assisted technique has been developed to assess absolute carotid plaque dimensions from B-mode ultrasonography, with enhanced capability of comparative examinations at large time intervals. Plaque area is measured from arterial lumen to adventitia with a real-time tissular detection program. Further measurements on the same patient are made using an echo-specific mask automatically generated by the computer from the original section. For an average sonographer, the manipulation takes no more than ten minutes for each view. In order to determine the reproducibility of this technique, a repeated measurement study (T0, T1, T2) was carried out on 8 patients with moderate to severe atherosclerotic lesions at carotid localizations. The plaque areas ranged from 52.7 to 202.3 mm2 (120.7 +/- 61). The coefficients of correlation between the measurements (T0-T1, T0-T2) were respectively 0.93 and 0.96 (P < 0.0001). The mean coefficient of variation (+/- SD) was 9.8% +/- 4.8. This study shows the feasibility of an accurate follow-up for atherosclerotic patients, with a two-dimensional plaque quantification, closer to the reality of the evolution of the pathology than the usual scoring system.

Aged↗

PubMed2004

[Genotyping and the quantification of hepatitis C virus by the melting curves in light cycler].

Detection of the HCV genome is crucial for diagnosis of HCV infection and for monitoring the efficacy of interferon treatment for patients with HCV. We developed a convenient screening test for HCV genotypes 1 and 2 based on the melting curve analysis with SYBER green I. Serum samples were drawn from 114 patients with known chronic HCV infection confirmed to be antibody-positive by immunoblot assay. A characteristic melting profile for each genotype was obtained by monitoring the fluorescence as the temperature increases through the melting point of the PCR product. Serum samples with HCV-RNA genotype (1b, 2a and 2b) were analyzed every test as standard samples and the genotype of unknown samples was determined by the comparison with the melting point of standard samples. Serum samples with known HCV-RNA genotype (1b, 2a and 2b) and HCV-RNA-negative sample were tested using the Light cycler system. The melting curve analysis indicated that melting points are 93.08 +/- 0.56 degrees C for genotype 1b (n = 63), 91.08 +/- 0.49 degrees C for genotype 2a (n = 33), and 91.77 +/- 0.28 degrees C for genotype 2b (n = 18). The melting points for genotypes 1b, 2a, and 2b differed by approximately 1 degree C in each other. The genotype was determined for all samples using Okamoto's method and Light cycler system, and both systems produced absolutely identical results for all the samples studied. Sixty-three of 114 were genotype 1b, 33 samples were genotype 2a, and 18 were genotype 2b. This melting curve analysis is a rapid and convenient screening test for differentiation of HCV genotypes 1 and 2.

Benzothiazoles↗

PubMed1981

Rapid analysis of human fecal bile acids.

A rapid, accurate, precise method for determining human fecal bile acids is reported. Feces are homogenized and then briefly extracted with boiling absolute ethanol. A portion of the extract is evaporated to dryness and the residue heated with mild alkali to hydrolyze bile acid 3 alpha-hydroxyl esters. Aliquots of hydrolyzed crude extract are treated with resazurin reagent which effects a series of enzyme catalyzed reactions in which bile acid free 3 alpha-hydroxyls are first oxidized to 3-oxo-groups in a reaction catalyzed by 3 alpha-hydroxysteroid dehydrogenase. Resulting protons are transferred to beta-nicotinamide adenine dinucleotide, yielding reduced beta-nicotinamide adenine dinucleotide (beta-NADH). beta-NADH then reduces nonfluorescent resazurin to fluorescent resorufin in a reaction catalyzed by diaphorase. Developed fluorescence, which is proportional to the extract aliquots bile acid content, is excited at 565 nm and read at 580 nm, wavelengths which lie in a spectral region in which there is minimal fecal pigment absorption. 3-Oxo-bile acids and bile acid 3 alpha-sulfates are extracted in the procedure but reduction and/or solvolysis is necessary before quantification.

Bile Acids and Salts↗

PubMed1999

Quantification in patient urine samples of felbamate and three metabolites: acid carbamate and two mercapturic acids.

PURPOSE: Previously we proposed and provided evidence for the metabolic pathway of felbamate (FBM), which leads to the reactive metabolite, 3-carbamoyl-2-phenylpropion-aldehyde. This aldehyde carbamate was suggested to be the reactive intermediate in the oxidation of 2-phenyl-1,3-propanediol monocarbamate to the major human metabolite 3-carbamoyl-2-phenylpropionic acid. In addition, the aldehyde carbamate was found to undergo spontaneous elimination to 2-phenylpropenal, commonly known as atropaldehyde. Moreover, atropaldehyde was proposed to play a role in the development of toxicity during FBM therapy. Evidence for atropaldehyde formation in vivo was reported with the identification of modified N-acetyl-cysteine conjugates of atropaldehyde in both human and rat urine after FBM administration. Identification of the atropaldehyde-derived mercapturic acids in urine after FBM administration is consistent with the hypothesis that atropaldehyde is formed in vivo and that it reacts with thiol nucleophiles. Based on the hypothesis that the potential for toxicity will correlate to the amount of atropaldehyde formed, we sought to develop an analytic method that would quantify the amount of relevant metabolites excreted in patient urine. METHODS: We summarize the results of an LC/MS method used to quantify FBM, 3-carbamoyl-2-phenylpropionic acid and two atropaldehyde-derived mercapturic acids in the patient population. RESULTS: Analysis was performed on 31 patients undergoing FBM therapy. The absolute quantities of FBM and three metabolites were measured. CONCLUSIONS: This method demonstrated sufficient precision for the identification of patients exhibiting "abnormal" levels of atropaldehyde conjugates and may hold potential for patient monitoring.

Acetylcysteine↗

PubMedDate not supplied

Expression of tetraspanin adaptor proteins below defined threshold values is associated with in vitro invasiveness of mammary carcinoma cells.

Tetraspanins are transmembrane adaptor proteins involved in the regulation of various fundamental cellular processes. For a number of malignant diseases, the level of expression of members of the tetraspanin family was found to correlate with tumor cell invasiveness, ability to form metastases, and poor clinical outcome. We describe the exact quantification of mRNAs coding for the tetraspanins CD9, CD63, CD82 and CD151 expressed by mammary carcinoma-derived cell lines that were classified as invasive or non-invasive according to their ability to penetrate collagen-fibroblast gels in vitro. The mean of beta2-microglobulin-normalized expression of CD9 was about 10-fold higher than the mean calculated for CD63 and about 20-fold higher than expression of CD82 and CD151. Direct comparison of tetraspanin expression of invasive and non-invasive cell lines with the Mann-Whitney test revealed a significant correlation for CD63. Grouping of cell lines in relation to threshold values of expression resulted in significant correlations for CD63 (Fisher's exact test p=0.004) and CD151 (p=0.02) but not for CD82 (p=0.065) and CD9 (p=0.168). Expression of CD9, C63 and CD151 was found to be coupled whereas CD82 was expressed independently. This highly significant association points to common mechanisms of gene regulation for this subgroup of tetraspanins. We showed that on basis of absolute amounts of tetraspanin mRNAs, at least in vitro invasiveness is clearly predictable. Our results support the assumption that downregulation of tetraspanins in breast cancer cells is an important step of tumor progression to more malignant phenotypes and underline their important role as mediators in multimolecular membrane protein complexes regulating cell adhesion and migration.

Antigens, CD↗

PubMed1994

Quantification of protein turnover in primary cultures of rat hepatocytes.

1. A radioactive tracer method, based on [3H]valine, for determination in parallel experiments of degradation of cellular proteins and synthesis of both cellular and secreted proteins in cultured rat hepatocytes was developed with special emphasis on methods of calculation, the number of protein pools and maintenance of nitrogen balance. 2. An extracellular concentration of 2 mM valine ensured a specific activity of the precursor pool for protein synthesis, which was constant during the experimental period and practically identical to that of extracellular valine. 3. Amino acid concentrations in the culture medium used were not rate-limiting for the synthesis of proteins. 4. The rate of labelling of the cellular protein pool during 8 days was in accordance with first-order saturation kinetics, which together with a constant ratio between labelling of soluble and membrane bound proteins, is compatible with a single pool of cellular proteins. 5. Protein degradation can be accurately measured by the release from prelabelled proteins of [3H]valine in the presence of 2 mM extracellular valine, if a 1 h chasing period is included in the experimental design. 6. The constancy of the degradation constant (kd) during protein labelling for up to 8 days, is in accordance with the existence of only one pool of cellular protein. 7. The cultured hepatocytes were in nitrogen balance during the experimental period of 8 days as reflected in a constant protein content per cell. The absolute rates of degradation and synthesis of cellular protein were identical, which confirm the validity of the method described.

Animals↗

PubMed2000

Measurement of extrastriatal D2-like receptor binding with [11C]FLB 457--a test-retest analysis.

[11C]FLB 457 is a radioligand for positron emission tomography (PET) that possesses high affinity to D2/D3 receptors. It has been suggested to be useful for quantification of low-density dopamine D2 receptor populations, e.g. in cortical and limbic brain areas. We explored the reproducibility of five methods for measuring extrastriatal D2-like receptor binding potential with [11C]FLB 457. Seven healthy male volunteers were examined twice with [11C]FLB 457 (high specific radioactivity) on the same day, at least 3 h apart. Four brain areas, frontal cortex, nucleus thalamus, temporal cortex and cerebellar cortex, were examined. Binding potentials (BPs) were derived from (1) a target to cerebellum distribution volume ratio, (2/3) two reversible reference tissue compartment models and (4) a transient equilibrium approach. For comparison, BP values were also calculated with the standard three-compartment kinetic model that does not assume a receptor-free reference region. The use of the standard three-compartment model did not result in reproducible BP estimates. The distribution volume (DV) ratio, reference tissue compartment models and the transient equilibrium method all had good to excellent intraclass correlation coefficients (ICCs) in the studied brain areas ranging from 0.56 to 0.93. Absolute variability was also relatively low, ranging from 5.3% to 10.4%. There were no marked differences in the ICC or absolute and relative variability between the four methods based on a reference tissue (cerebellum). In addition, we did not observe systematic differences in the BP between the first and the second scan. These data indicate that the reproducibility of the DV ratio, reference tissue models and the transient equilibrium method is good or excellent. However, each of these methods includes assumptions affecting their validity. Thus, the choice of method will be critically dependent on the purpose of the study.

Adult↗

PubMed2005

Power and sample size calculations in the presence of phenotype errors for case/control genetic association studies.

BACKGROUND: Phenotype error causes reduction in power to detect genetic association. We present a quantification of phenotype error, also known as diagnostic error, on power and sample size calculations for case-control genetic association studies between a marker locus and a disease phenotype. We consider the classic Pearson chi-square test for independence as our test of genetic association. To determine asymptotic power analytically, we compute the distribution's non-centrality parameter, which is a function of the case and control sample sizes, genotype frequencies, disease prevalence, and phenotype misclassification probabilities. We derive the non-centrality parameter in the presence of phenotype errors and equivalent formulas for misclassification cost (the percentage increase in minimum sample size needed to maintain constant asymptotic power at a fixed significance level for each percentage increase in a given misclassification parameter). We use a linear Taylor Series approximation for the cost of phenotype misclassification to determine lower bounds for the relative costs of misclassifying a true affected (respectively, unaffected) as a control (respectively, case). Power is verified by computer simulation. RESULTS: Our major findings are that: (i) the median absolute difference between analytic power with our method and simulation power was 0.001 and the absolute difference was no larger than 0.011; (ii) as the disease prevalence approaches 0, the cost of misclassifying a unaffected as a case becomes infinitely large while the cost of misclassifying an affected as a control approaches 0. CONCLUSION: Our work enables researchers to specifically quantify power loss and minimum sample size requirements in the presence of phenotype errors, thereby allowing for more realistic study design. For most diseases of current interest, verifying that cases are correctly classified is of paramount importance.

Alzheimer Disease↗

PubMed1995

Evaluation of technical factors affecting the quantification of trabecular bone structure using magnetic resonance imaging.

High resolution magnetic resonance (MR) techniques combined with standard techniques of stereology and texture analysis have been used to quantify trabecular structure. Using dried excised specimens from the tibia (n = 10) and radius (n = 2) we evaluate the impact of using volumetric gradient-echo (GE) and spin-echo (SE) MR imaging sequences, the relative importance of echo time in gradient-echo MR imaging, and the impact of different threshold values to segment the bone and bone marrow on the estimation of trabecular bone structure. We also investigate the inter-relationships between the different structural parameters derived from MR images. Images were obtained using fast gradient-echo and spin-echo imaging sequences, with TE values ranging from 7 to 17 ms using 4.7 and 1.5 Tesla imaging systems. In-plane image resolution ranged from 128 to 156 microns, and slice thickness ranged from 128 to 1000 microns. We derived stereological measures such as the mean intercept length, trabecular width, fractional area of trabecular bone, trabecular number, and trabecular spacing, the fractal dimension as a texture-related parameter and the Euler number as a measure of connectivity from these images. We found that the mean intercept length as a function of angle traced an ellipse with the orientation of the principal axis of the ellipse, a measure of trabecular orientation, identical when measured from the spin-echo or gradient-echo MR images. Absolute measures such as the fractional area, trabecular width, trabecular number, and fractal dimension as measured from gradient echo images were 28%, 30%, 1.3%, and 0.6% greater, respectively, than those calculated from spin-echo images, while the trabecular spacing was 14% less when calculated from gradient-echo images compared to spin-echo images. The structural parameters also depended on the echo time used to obtain the MR image. The choice of the threshold used to segment the high resolution images also affected the estimated structural parameters significantly. Our results indicate that MR may be used to visualize and quantify trabecular bone architecture; however, the different technical factors that could affect the appearance of MR images must be understood and considered in the data analysis and interpretation.

Absorptiometry, Photon↗

PubMed1996

Multicenter comparison of three commercial methods for quantification of human immunodeficiency virus type 1 RNA in plasma.

Three procedures for the quantification of human immunodeficiency virus type 1 (HIV-1) RNA from plasma were compared at three laboratories. The comparison involved the Quantiplex branched DNA assay (version 1.0) by Chiron Diagnostics, the NASBA-QT assay by Organon Teknika, and the Amplicor Monitor assay by Roche Molecular Systems. The laboratories performed each of the three assays with the same sets of reconstructed HIV-1-infected human plasma samples, cross-sectionally collected clinical plasma samples and longitudinally collected plasma samples from patients starting zidovudine therapy. Analysis of the reconstruction panel results for interlaboratory variation demonstrated that no laboratory differences in results were detected for any of the assays. A comparison of the reproducibilities of duplicate samples analyzed by batch and in separate assay runs demonstrated that the reproducibilities of the test results were similar within one assay and appeared to be independent of the HIV-1 concentration. The best reproducibility was obtained with the Quantiplex assay, but all three assays demonstrated equal reliability, which was independent of batched or unbatched analysis of replicate samples. Differences in the absolute concentrations calculated were observed for the assays, in particular in the analysis of reconstructed samples. In all assays, similar changes in plasma HIV-1 RNA concentrations were determined for longitudinally collected clinical samples.

Biomarkers↗

PubMed2000

Cardiac output quantification by Doppler echocardiography in intensive care--limitations and validation.

UNLABELLED: Cardiac output (CO) quantification is primordial to the evaluation of patients with heart failure who are on tailored therapy and under invasive hemodynamic monitoring. Doppler echocardiography can be used to access CO noninvasively, but the concordance between its results and those obtained by invasive methods in paired measurements is still controversial. To our knowledge, no previous studies have assessed the clinical relevance of Doppler echocardiography for CO serial evaluation in patients submitted to tailored therapy. AIM: To evaluate the usefulness of echo-Doppler in the assessment of CO and quantification of changes in CO, compared to thermodilution, in patients with advanced heart failure under hemodynamic monitoring to guide tailored therapy. METHODS: In 20 patients (14 male, 62 +/- 14 years old, all in sinus rhythm), with dilated cardiomyopathy and NYHA IV, admitted to the intensive care unit (ICU), CO was simultaneously determined by Doppler echocardiography (dpCO) and thermodilution (tdCO) in three serial evaluations (overall 60). The dpCO was calculated by multiplying the aortic orifice area by the velocity-time integral of aortic continuous wave Doppler flow and by the heart rate. A difference between tdCO and dpCO of more than 20% was considered a major error. RESULTS: In the overall evaluations, dpCO systematically overestimates tdCO (p = 0.026). The correlation between tdCO and dpCO was 0.81, the mean difference between measurements was 0.40 +/- 0.61 l/min (mean -2SD = -1.62 mean +2SD = 0.81) and 19 (32%) major errors occurred. No significant difference was found between CO percentual variation values assessed by both methods, with a stronger correlation (r = 0.92-p = 0.014) compared to that found for absolute values. On using the dpCO/tdCO ratio in the first evaluation to correct subsequent dpCO, the correlation was fairly good (r = 0.96-p = 0.0002 versus corrected dpCO). The mean difference between paired measurements was significantly lower (0.12 +/- 0.28 l/min-mean 2SD = -0.44 mean +2SD = 0.67), and there were no errors. CONCLUSIONS: CO estimated by Doppler echocardiography has a good correlation with thermodilution although with a weak concordance between paired results in patients with dilated cardiomyopathy and advanced heart failure admitted to the ICU for tailored therapy. Our results with dpCO percentual change in repeated evaluations and with corrected dpCO value after a single simultaneous invasive determination suggest that Doppler echocardiography is a valid method for clinical purposes, allowing us to propose a reduction in the time period of invasive hemodynamic monitoring.

Adult↗

PubMed2005

Quantification of tension in Thera-Band and Cando tubing at different strains and starting lengths.

AIM: Elastic resistance exercise is frequently used in rehabilitation programs. Precise quantification using color coded bands is not possible. A method to predict forces for any length of mostly used elastic tubings and thereby a general equation for the comparison of elastic material is presented. METHODS: In an in vitro-series the tubing tension in relation to elongation of 6 colors of Thera-Band and Cando tubings was directly measured. The relation between tension and elongation was determined depending on different resting lengths. In clinical practice tension and elongation are prescribed. Two mathematical methods to calculate the resting length as regulatory variable are presented. One is based on the construction of iso-tension curves, the 2nd on the general mathematical relation delta % tension=K* % length change+n. RESULTS: 1. The tension/elongation relation is linear beyond a threshold in the early part of elongation. 2. The iso-tension method works graphically and facilitates the precise determination of resting length. 3. The double % relation between tension and elongation method describes the general properties of all tubings, where K=0.75 is a general constant. To calculate absolute values for the resting length, corresponding to a given tension and elongation, special ''color constants'' are presented. CONCLUSIONS: The prediction of maximum tension values for any given elongation is possible by simple variation of resting length. These findings may improve the usefulness of elastic tubings in rehabilitation programs. The mathematical method further may be used for the comparison of elastic materials with linear characteristics.

Algorithms↗

PubMed2004

Quantification of alpha-subunit isoforms of Na,K-ATPase in rat resistance vessels.

AIM: Rat mesenteric resistance vessels (RV) were characterized with respect to concentration of individual alpha-subunit isoforms of Na,K-ATPase. METHODS: Total vessel homogenates were used to avoid any loss or subfractionation of membranes. They were applied to sodium dodecyl sulphate gels and, for calibration, in parallel lanes were run purified rat Na,K-ATPase preparations with known isoform distribution and content. The capacity per mg protein for Na+-dependent 32P-phosphorylation of Na,K-ATPase isolated from rat kidney was used for alpha1 calibration and that for high-affinity (3H)ouabain binding of Na,K-ATPase isolated from rat brain was used for (alpha2 + alpha3) calibration. Western blots containing homogenate proteins and reference enzyme were incubated with isoform-specific antibodies and radiolabelled secondary antibodies. The signals from adjacent alpha spots were used for qualitative and quantitative characterization of rat vessels. RESULTS: A concentration of 100.7 +/- 14.4 pmol (n = 11) per g wet weight of the alpha1-isoform containing Na,K-ATPase was found in RV from 12-14-week rats. A much lower and more unreliable content of alpha2- and alpha3-isoforms was found. These ouabain-sensitive isoforms seem to represent a maximum of 5-10% each compared with the ouabain-insensitive rat alpha1-isoform. CONCLUSIONS: The isoform pattern in RV, in which the isoform with high/intermediate Na+-affinity is the absolutely dominating one representing nearly all sodium pumps in this tissue, is very different from that seen in rat skeletal muscles. Due to the high content of the ouabain-insensitive alpha1-isoform in rat RV this species would seem a less relevant model in studies addressing a role of cardiac glycosides and putative endogenous ouabain-like factors in hypertension.

Animals↗

PubMed2001

Rare-event analysis of circulating human dendritic cell subsets and their presumptive mouse counterparts.

BACKGROUND: Considerable interest has focused recently on murine CD8alpha- and CD8alpha+ dendritic cell (DC) subsets, because of their roles in initiating and regulating immune responses. Attention has also centered on their presumed human counterparts, DC1 and DC2, respectively, and their precursors. Identification and quantification of these subsets in the blood may be crucial to understanding and monitoring of their immunologic significance, particularly in humans, where blood may be the only tissue readily or routinely available. METHODS: Leukocytes were isolated from anticoagulated human or mouse (C57BL/10J) blood using conventional procedures. Four-color, rare-event, flow cytometric analysis was used to identify DC1 precursors (pDC1; lineage [lin]- CD4+ CD11c+ HLA-DR+) or DC2 precursors (pDC2; lin- CD4+ CD11c- CD123(hi) [IL-3Ralpha(hi)] HLA-DR+) in normal humans. In mice, CD8alpha+ (CD11b(lo), CD11c+) and CD8alpha- (CD11b(hi), CD11c+) DC subsets were identified both in normal animals and after administration of the potent DC growth factor, fms-like tyrosine kinase 3 ligand (Flt3L). RESULTS: All human subjects examined had discrete populations of pDC1 and pDC2 comprising approximately 0.6% and 0.1% of blood mononuclear cells. CD8alpha- and CD8alpha+ DC constituted approximately 0.75% and 0.2%, respectively, of blood mononuclear cells in normal mice, and 12% and 0.5%, respectively, in Flt3L-treated animals. Flt3L administration substantially increased the absolute numbers of circulating CD11c+ DC by approximately 200-fold. CONCLUSIONS: In addition to pDC1 and CD8alpha- DC, pDC2 and CD8alpha+ DC can be identified in normal human or mouse blood, respectively. Monitoring and isolation or characterization of these cells may provide novel insights into their functional significance in transplantation and other clinical conditions.

Adjuvants, Immunologic↗

PubMed2006

[Pregnancy-associated venous thromboembolic disease: prediction, prevention, and therapy].

Thromboembolic disease remains a leading cause of maternal mortality during pregnancy and the puerperium. Rational and risk-adapted administration of heparin prophylaxis depends on 1. the identification of those women who have an increased risk of thrombosis and 2. the accurate quantification of this risk. In women without prior thrombosis, the presence of a heterozygous factor V Leiden or heterozygous G20210A mutation in the prothrombin gene is associated with a pregnancy-associated thrombotic risk of approximately 1 in 400. Thus, in pregnant carriers of either one of these mutations the risk of venous thromboembolism is low. Therefore, no heparin prophylaxis is recommended. A combination of the two genetic risk factors can increase the risk to a modest level of 1 in 25. In women with a single episode of prior thrombosis associated with a transient risk factor, e.g. surgery or trauma, and no additional genetic risk factor, the probability of a pregnancy-associated thrombosis appears also to be low. However, data are sparse and conflicting. In contrast, in women with a prior idiopathic venous thrombosis who carry an additional hereditary risk factor or who have a positive family history of thrombosis, a high risk (>10%) can be expected supporting the indication for active antepartum and postpartum heparin prophylaxis. Despite the remarkable progress in risk stratification, the absolute magnitude of risk and the optimal management in many cases is an issue of ongoing debate.

Antiphospholipid Syndrome↗

PubMed2003

Automated measurement of microaneurysm turnover.

PURPOSE: An automated system for the measurement of microaneurysm (MA) turnover was developed and compared with manual measurement. The system analyses serial fluorescein angiogram (FA) or red-free (RF) fundus images; fluorescein angiography was used in this study because it is the more sensitive test for MAs. Previous studies have shown that the absolute number of MAs observed does not reflect the dynamic temporal nature of the MA population. In this study, almost half of the MAs present at baseline had regressed after a year and been replaced by new lesions elsewhere. METHODS: Two clinical datasets were used to evaluate the performance of the automated turnover measurement system. The first consisted of 10 patients who had two fluorescein angiograms acquired a year apart. These data were analyzed, both manually and using the automated system, to investigate the inter- and intraobserver variations associated with manual measurement and to assess the performance of the automated system. The second dataset contained FAs from a further 25 patients. This dataset was analyzed only with the automated system to investigate some properties of microaneurysm turnover, in particular the differing detection sensitivities of new, static and regressed microaneurysms. RESULTS: Manual measurements exhibited large inter- and intraobserver variation. The sensitivity and specificity of the automated system were similar to those of the human observers. However, the automated measurements were more consistent-an important condition for accurate turnover quantification. Regressed MAs were more difficult to detect reliably than new MAs, which were themselves more difficult to detect reliably than static MAs. CONCLUSIONS: The automated system was shown to be fast, reliable, and repeatable, making it suitable for processing large numbers of images. Performance was similar to that of trained manual observers.

Adult↗