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Results for “Absolute quantification”

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PubMed1999

Quantification of dynamic changes in cerebral venous oxygenation with MR phase imaging at 1.9 T.

A double-echo magnetic resonance (MR) phase imaging method was used to measure the time-dependent changes in regional cerebral venous blood oxygenation during neural stimulation. Our results showed that a finger opposition task induced an absolute increase of blood oxygenation level of deltaY = 0.16+/-0.05 (n = 6), which was measured in the small veins located in the left central sulcus. It was observed that there is a long temporal delay of approximately 27 sec after the onset of the task to reach the steady-state venous oxygenation. This work demonstrated that MRI is a powerful clinical tool for in vivo monitoring of the dynamic nature of organ-specific venous blood oxygenation.

Adult↗

PubMed1994

Quantification of regional cerebral blood flow and volume with dynamic susceptibility contrast-enhanced MR imaging.

PURPOSE: Quantification of regional cerebral blood flow (rCBF) and volume (rCBV) with dynamic magnetic resonance (MR) imaging. MATERIALS AND METHODS: After bolus administration of a paramagnetic contrast medium, rapid T2*-weighted gradient-echo images of two sections were acquired for the simultaneous creation of concentration-time curves in the brain-feeding arteries and in brain tissue. Absolute rCBF and rCBV values were determined for gray and white brain matter in 12 subjects with use of principles of the indicator dilution theory. RESULTS: The mean rCBF value in gray matter was 69.7 mL/min +/- 29.7 per 100 g tissue and in white matter, 33.6 mL/min +/- 11.5 per 100 g tissue; the average rCBV was 8.0 mL +/- 3.1 per 100 g tissue and 4.2 mL +/- 1.0 per 100 g tissue, respectively. An age-related decrease in rCBF and rCBV for gray and white matter was observed. CONCLUSION: Preliminary data demonstrate that the proposed technique allows the quantification of rCBF and rCBV. Although the results are in good agreement with data from positron emission tomography studies, further evaluation is needed to establish the validity of method.

Brain↗

PubMed1995

Accuracy of urea removal estimated by kinetic models.

The most accurate method for assessing the dialysis dose delivered during high efficiency/flux hemodialysis has not been established. Most current indices of dialysis dose are based on blood-side urea measurements, and thus estimate urea removal. Unfortunately, these methods may lead to inappropriately short dialysis during high flux or high efficiency dialysis, perhaps because of inaccuracies in estimating the amount of urea removal. It is unknown whether these clearance-based approaches can accurately predict either absolute or fractional net urea removal, the latter being equivalent to the solute removal index (SRI). Therefore, we compared the urea removal calculated by five blood-side kinetic methods: (1) urea reduction ration, (2) 1-pool, (3) 2-pool models, and the (4) Smye and (5) Daugirdas formulae. These were compared with the gold standard measurement by direct dialysate quantification. Eight stable patients receiving high-flux hemodialysis were studied over four sessions each. BUN was measured at 0, 45 minutes, 90 minutes, end dialysis, one hour after dialysis (equilibrium value), and 48 hours later. Total body water was determined from the dialysate urea removal; the urea generation rate was calculated using one hour post-dialysis and 48-hour BUN values. Both the total body water and urea generation rate were provided to the 1- and 2-pool models to optimize accuracy. The urea reduction ratio overestimated SRI. The 1-pool model overestimated both absolute urea removal and SRI in 28 of 32 sessions. The 2-pool model slightly underestimated both absolute urea removal and SRI. In contrast, the Smye and Daugirdas formulas accurately estimated SRI.(ABSTRACT TRUNCATED AT 250 WORDS)

Female↗

PubMed2004

Quantification of oxygen fluxes in a long gravity sewer.

Quantification of the oxygen fluxes in the sewer system is at present the optimal methodology to obtain information about the influence of sewers on transformations and mass balances in the urban drainage system. However, the relative and absolute values of these fluxes are practically unknown. In this work, the oxygen fluxes were quantified experimentally in a full-scale aerobic main sewer. The sewer biofilm respiration was determined with an in situ flow cell, a method that has not been used before in the sewer. The surface reaeration was determined with a gas tracer method based on the inert, non-radio-active and non-toxic gas tracer sulphur hexafluoride. In addition, the wastewater biomass respiration rate was measured. The validity of the applied methods was verified with redundant oxygen balances over a 2-km-long section. Measurement campaigns under different hydrodynamic conditions showed that the relative contribution of the biofilm, the wastewater, the reaeration and the in- and outflow with the water, all contributed significantly. However, the absolute contributions varied extensively and depended especially on the discharge. The COD conversion in the sewer could be estimated from the aerobic activity. The aerobic total degradation in the study reach was 3%. However, when extrapolated to the entire sewer net of the catchment area with 5000 PE, the COD conversion was estimated as high as 30% of the dissolved COD during the night. This indicates that the wastewater composition at the treatment plant will be strongly affected by the sewer system.

Bacteria, Aerobic↗

PubMed2000

Normalizing complementary DNA by quantitative reverse transcriptase-polymerase chain reaction of beta2-microglobulin: molecular monitoring of minimal residual disease in acute promyelocytic leukemia.

Reverse transcription (RT)-polymerase chain reaction (PCR) raises unique methodological matters that may hamper the reliability of the procedure, especially when results should direct therapeutic decisions. One of these matters is represented by the RT step. The present study shows that differences in complementary DNA (cDNA) preparations purposely containing increasing amounts of retrotranscribed RNA were not disclosed by nonquantitative RT-PCR by two different housekeeping genes, leading to fictitious results when the expression of a given gene was quantitatively assessed. To overcome this problem, the following are proposed: 1) to evaluate the efficiency of RT step through the quantification, by competitive RT-PCR, of the expression levels of the housekeeping gene beta2-microglobulin (beta2M); 2) to normalize each cDNA preparation to be comprised within 1 standard deviation of the mean value of beta2M absolute level (3.14 +/- 1.14 attomoles/microg RNA) found by analyzing 33 cell lines of hematopoietic origin. To validate this strategy in a clinical setting, serial cDNA samples from patients were checked by conventional and quantitative RT-PCR for beta2M. Again, only a quantitative evaluation of beta2M levels was allowed to unveil significant differences, otherwise undetected, in the efficiency of RT reactions among these cDNA samples. Normalization of samples to obtain cDNA preparations containing comparable beta2M levels, eventually led to an increased sensitivity in the detection of PML-RARalpha fusion transcripts. This approach seems of great value for the monitoring of minimal residual disease in serial patient samples when a tumor-specific marker is available.

Actins↗

PubMed2005

A method for aligning trans-tibial residual limb shapes so as to identify regions of shape change.

Quantification of the change in shape of a residual limb over time is relevant to the fitting of an external prosthesis. Three algorithms were developed and evaluated to align residual limb shapes: iterative closest points (ICP), mean absolute difference, and weighted surface normals/mean absolute difference. Evaluations were conducted by aligning residual limb shapes with known deformations and transformations with their original shapes. Results showed that ICP did not perform well in that it tended to favor a global distribution of local shape difference rather than localization of the error. The mean absolute difference algorithm performed well as long as the shape difference was localized to one region. Weighted mean surface normals/absolute difference provided the best alignment results, performing well both if shape changes were localized and if they were globally distributed. Mean alignment errors for this method were less than 0.285 mm for each of the three translation directions and less than 0.357 degrees for each of the three rotation directions. This algorithm could be helpful to patients, prosthetists, and researchers developing treatments to overcome the detrimental fitting effects of residual limb shape change.

Algorithms↗

PubMed2006

Quantification of estrogenic mycotoxins at the ng/L level in aqueous environmental samples using deuterated internal standards.

Because of their pronounced estrogenicity, resorcyclic acid lactones (RALs) are of concern in aqueous environments even at the low ng/L level. Therefore, we developed an accurate, precise and sensitive HPLC-MS/MS method to detect these mycotoxins in different aqueous environmental samples. The compounds investigated included zearalenone (ZON), alpha- and beta-zearalenol, zearalanone as well as alpha- and beta-zearalanol. The use of isotope labelled internal standards (in this case deuterated RAL-analogues) ensured an accurate quantification of the target analytes, independent of matrix compounds interfering with the analytes during ionisation and analyte losses occurring during sample preparation. Sample enrichment was carried out by solid-phase extraction (SPE) using Supelclean Envi-18 cartridges. Absolute method recoveries for all analytes ranged from 95 to 108%, 70 to 102%, and 76 to 109%, method detection limits from 0.5 to 2.1 ng/L, 0.4 to 1.1 ng/L, and 0.8 to 12.4 ng/L and precision from 3 to 14%, 2 to 13% and 4 to 16% in drainage water, river water and wastewater treatment plant (WWTP) effluent, respectively. The method was applied to verify the emission of RALs from a Fusarium graminearum infested crop field into the drainage system. Zearalenone was present in drainage water in concentrations up to 30 ng/L. So far, none of the other five investigated compounds have been detected.

Analytic Sample Preparation Methods↗

PubMed2006

Use of piezoelectric-excited millimeter-sized cantilever sensors to measure albumin interaction with self-assembled monolayers of alkanethiols having different functional headgroups.

In this paper, we describe a new modality of measuring human serum albumin (HSA) adsorption continuously on CH3-, COOH-, and OH-terminated self-assembled monolayers (SAMs) of C11-alkanethiols and the direct quantification of the adsorbed amount. A gold-coated piezoelectric-excited millimeter-sized cantilever (PEMC) sensor of 6-mm2 sensing area was fabricated, where resonant frequency decreases upon mass increase. The resonant frequency in air of the detection peak was 45.5 +/- 0.01 kHz. SAMs of C11-thiols (in absolute ethanol) with different end groups was prepared on the PEMC sensor and then exposed to buffer solution containing HSA at 10 microg/mL. The resonant frequency decreased exponentially and reached a steady-state value within 30 min. The decrease in resonant frequency indicates that the mass of the sensor increased due to HSA adsorption onto the SAM layer. The frequency change obtained for the HSA adsorption on CH3-, COOH-, and OH-terminated SAM were 520.8 +/- 8.6 (n = 3), 290.4 +/- 6.1 (n = 2), and 210.6 +/- 8.1 Hz (n = 3), respectively. These results confirm prior conclusions that albumin adsorption decreased in the order, CH(3) > COOH > OH. Observed binding rate constants were 0.163 +/- 0.003, 0.248 +/- 0.006, and 0.381 +/- 0.001 min(-1), for methyl, carboxylic, and hydroxyl end groups, respectively. The significance of the results reported here is that both the formation of self-assembled monolayers and adsorption of serum protein onto the formed layer can be measured continuously, and quantification of the adsorbed amount can be determined directly.

Adsorption↗

PubMed2004

TaqMan real-time polymerase chain reaction assay for the correlation of Treponema denticola numbers with the severity of periodontal disease.

Treponema denticola has been implicated in periodontitis, and the presence of this organism in periodontal pockets has been investigated. However, qualitative analysis is insufficient for the clinical evaluation of periodontal treatments, and quantification of T. denticola populations is essential for monitoring therapeutic efficacy. Therefore, we developed a quantitative method for T. denticola that uses the TaqMan real-time polymerase chain reaction assay. Using this system, we evaluated the relative and absolute numbers of this organism in saliva and subgingival plaque. Furthermore, we analyzed the relationship between the numbers of T. denticola and pocket depth, and found a significant positive correlation (P < 0.0001) between these parameters. This report demonstrates the broad potential of real-time polymerase chain reaction applications in periodontology.

Adult↗

PubMedDate not supplied

HPLC determination of amoxicillin comparative bioavailability in healthy volunteers after a single dose administration.

PURPOSE: An accurate, precise and sensitive HPLC assay was developed for the determination of amoxicillin in human plasma samples, to compare the bioavailability of two amoxicillin capsule (500mg) formulations (Amoxicilina from Brazil, as a test formulation and Amoxil from SmithKline Beecham Laboratories Ltda., Brazil, as a reference formulations) in 24 volunteers of both sexes. METHODS: Amoxicillin concentrations were analyzed by combined reversed phase liquid chromatography and UV detection (lambda=229 nm). Amoxicillin and cefadroxil (internal standard) were extracted from the plasma by addition of cold methanol. The separation was achieved using the Lichrosorb 10 microm, C18 reversed phase column at room temperature. The mobile phase consisted of a 95% phosphate buffer (0.01 mol/L), pH=4.8 and 5% acetonitrile mixture. The study was conducted using an open randomized 2-period crossover balanced design with a 1-week washout period between the doses. Plasma samples were obtained over an 8-hour period. The bioequivalence between the two formulations was assessed by calculating individual peak plasma concentrations (C(max) ) and area under the curve (AUC(0-8h) ) ratios (test/reference). The statistical interval proposed was 80-125%, as established by the US Food and drug administration Agency. RESULTS: The internal standard and amoxicillin eluted about 4.2 and 5.2 min, respectively at a flow rate of 1.3ml/min. The mean absolute recovery of AMO in plasma was 90.0% at 3 microg/ml, 98.6% at 25 microg/ml and 95.3 at 50 microg/ml. The assay showed excellent relationships between peak height ratios and plasma concentrations (r(2)>or= 0.999). The limit of quantification was 1g/ml, based on 200l of plasma. The geometric mean of Amoxicilina/Amoxil 500 mg capsules individual percentage ratio was 101.4% for AUC(0-8h), and 99.9% for C(max). The 90% confidence intervals were 98.3-104.4% and 95.7-103.9%, respectively. CONCLUSION: This simple, rapid and selective method is suitable for pharmacokinetic, bioavailability and bioequivalence studies. Since the 90% CI for both C(max )and AUC(0-8h) lies within the 80-125% interval proposed by the Food and Drug Administration, it was concluded that Amoxicilina 500 mg capsules was bioequivalent to Amoxil capsules 500 mg, in terms of both the rate and extent of absorption.

Amoxicillin↗

PubMed1998

Quantification of the nucleocytoplasmic distribution of wild type and modified proteins using confocal microscopy: interaction between 90-kDa heat shock protein (Hsp90 alpha) and glucocorticosteroid receptor (GR).

The investigation of molecular interactions in whole cells by immunofluorescence was developed recently, based on the targeting of the protein partners to different cellular compartments and analysis of the modifications in their subcellular distribution resulting from their interaction. This paper describes the adaptation of the confocal microscopy to the quantification of the partitioning of transiently coexpressed proteins between nucleus and cytoplasm. We defined a nucleocytoplasmic ratio R, corresponding to the difference between nuclear and cytoplasmic fluorescence intensities divided by their sum (N - C/N + C), which does not refer to absolute fluorescence intensities. Interaction was detected by statistically comparing the distribution of R value frequencies in cell populations expressing one or both proteins. The convenience of this whole cell method was demonstrated by detecting and analyzing interaction between the human glucocorticosteroid receptor (GR) and the chick 90-kDa heat shock protein (Hsp90), using various combinations of wild-type and nuclear- or cytoplasmic-targeted GR and Hsp90. In addition, three Hsp90 deletion/ truncation mutants were tested: the C-terminal truncated mutant NC4 interacted slightly, indicating the contribution of this part of the molecule to the interaction with GR, while the shorter truncated mutant NC6 did not interact with GR, likely resulting from an incorrect folding of the molecule. No role for the first charged region (delta A') was found as shown by the strong interaction detected for the delta A'Hsp90. This method can fruitfully be applied to the delimitation of the amino-acid sequences involved in protein-protein interaction by mutational analysis, especially to seek confirmation of other methods or when other approaches have failed.

Animals↗

PubMed1984

Bronchoalveolar lavage in the normal lung. 2. Cell distribution and cytomorphology.

Bronchoalveolar lavage, performed on 15 healthy volunteers, enabled quantification and characterization of the alveolar cell populations. The subjects studied were 8 nonsmokers (5 males, 3 females) and 7 smokers (6 males, 1 female). It was found that in the smokers the macrophages increased compared with nonsmokers, both in absolute number (419,000 vs. 138,000/ml; p less than 0.005) and in percentage (93.8 +/- 3.0 vs. 88.1 +/- 4.8%; p less than 0.02), causing a significant increase in the total number of cells recovered after bronchoalveolar lavage (471,000 vs. 163,000/ml; p less than 0.005). Lymphocytes and neutrophils do not significantly vary in the two groups, even though among the smokers there is a tendency for the concentration of these cells to increase in the lavage liquids. The importance of the data obtained from healthy subjects lies in the possibility thus afforded of having reference values for the study of various lung pathologies with bronchoalveolar lavage.

Adult↗

PubMed1995

Human immunodeficiency virus-related microvasculopathy and Kaposi's sarcoma: a case-control study.

Ocular microangiopathic syndrome including retinal and conjunctival abnormalities is frequently found in patients with human immunodeficiency virus type 1 (HIV-1) disease. Kaposi's sarcoma (KS) is the most frequent neoplasia found in patients with HIV-1 disease. We have recently reported a significant association between conjunctival microvasculopathy and KS in 117 patients with HIV-1 disease. The objective of the present study was to determine whether this association is existent when matched patients with and without KS are compared. A total of 22 matched pairs were obtained under consideration of the absolute CD4+ lymphocyte count, Walter Reed (WR) classification, gender, and serum levels of beta-2-microglobulin and neopterin. Conjunctival microangiopathy was determined for each eye by a standardized rating scale ranging from 0 to 5, allowing a reliable and valid quantification of conjunctival blood-flow sludging. The mean value obtained for conjunctival sludge was 1.8 (SEM, 0.4) for patients without KS and 3.2 (SEM, 0.3) for patients with KS, demonstrating a clinically and statistically significant difference between the two groups (Student's t = 3.0; P = 0.003). This difference was higher when patients with a CD4+ lymphocyte count exceeding 200/microliters were regarded. Similar factors or mechanisms may contribute to HIV-related conjunctival microvasculopathy and KS.

Adult↗

PubMed1988

Quantification of regional myocardial function by rapid cine MR imaging.

The capability of rapid (cine) MR imaging to quantitate left ventricular function was assessed in 13 normal subjects and in 15 patients with ischemic heart disease and regional wall-motion abnormalities proved by echocardiography and/or by contrast ventriculography. Fifteen to 20 MR images/cardiac cycle were acquired by using partial flip angles, short repetition times, and gradient-refocused echoes. Regional wall motion was assessed qualitatively in the equatorial left ventricular section by using the cine display and quantitatively by measuring myocardial thickness at end-diastole and at end-systole in six left ventricular segments in this plane. In normal volunteers wall motion was normal in all segments. Heterogeneity of systolic wall thickening was observed in normal subjects, ranging from 33% +/- 17% in the posteroseptal segment to 66% +/- 29% in the posterior segment. Overall systolic wall thickening was 48% +/- 28%. From the cinematic display of MR images, abnormal wall motion was observed in 40 of 90 segments in patients with ischemic heart disease, which correlated well with results of echocardiography or contrast ventriculography. Twenty-one segments were hypokinetic, 15 were akinetic, and four were dyskinetic. In patients with ischemic heart disease, percentage systolic wall thickening was 43% +/- 31% in the segments with normal wall motion, 6% +/- 18% in hypokinetic segments, -4% +/- 24% in akinetic segments, and -13% +/- 25% in dyskinetic zones. Absolute systolic wall thickening was less than 2 mm in 31 of 40 abnormal segments and was greater than 2 mm in only three. Rapid acquisition, improved temporal resolution, and the capacity for cine display make this new MR technique potentially useful not only for qualitative assessment of cardiac wall motion, but also for quantification of regional myocardial function.

Adult↗

PubMed2006

A confirmatory analysis of malachite green residues in rainbow trout with liquid chromatography-electrospray tandem mass spectrometry.

A quantitative liquid chromatography-tandem mass spectrometric (LC-MS/MS) method has been developed for the determination of malachite green (MG) and its metabolite leucomalachite green (LMG) in fish. Residues were extracted with an acetonitrile-acetate buffer and purified using the automated solid-phase extraction (ASPEC). Residues were analyzed with a reversed-phase LC-MS/MS using a positive-ion electrospray ionisation (ESI). Isotope-labelled leucomalachite green (LMG-D5) was used as an internal standard for the quantification of LMG residues. The related dye, brilliant green (BG) was used as an instrumental standard. Identification and quantification of analytes were based on the ion transitions monitored by multiple reaction monitoring (MRM). The decision limit (CCalpha) for MG and LMG was 0.13 and 0.16 microgkg(-1). The respective detection capabilities (CCbeta) were 0.22 and 0.27 microgkg(-1). The absolute recovery (repeatability SD(r)) was in the range of 58-65% (7.8-11.2%) for MG and 59-68% (9.7-16.9%) for LMG. LMG was quantified also based on the internal standard, giving a recovery (repeatability SD(r)) of 103-110% (4.8-9.3%). The method was further evaluated by analyzing a total of 34 fish residue monitoring samples, of which eight samples were found to be non-compliant containing low residues of LMG.

Animals↗

PubMed1974

[The use of stereologic image analysis in medicine (peripheral nerves)].

The use of image plane scanning for the purpose of quantification of data is becoming increasingly important in medicine and biology. Such equipment allows complex geometric-statistical relationships to be investigated by reduction to a counting process in which picture elements are counted (using a photocell), stored, and evaluated. Various absolute values of interest can then be computed using certain mathematical relationships. These methods, using automatically controlled sampling, are fast, and, since they are able to detect small variations in light intensity, very accurate. Furthermore, even in the presence of highly complex patterns, an accurate surface area measurement can be made. Peripheral nerve cells, for example, lend themselves well to such methods, since the strong contrast between individual structural elements provides excellent conditions for automatic measurement.

Axons↗

PubMedDate not supplied

Quantification of statistical type I and II errors in correlation analysis of simulated functional magnetic resonance imaging data.

The potential of statistical analyses of functional magnetic resonance images using various threshold strategies in combination with correlation analysis was studied by simulating brain activation. Differences in statistical Type I (alpha) and II (beta) errors are substantial for the various thresholds. Absolute thresholds and individualized thresholds based on the assumption of a gaussian noise distribution are producing constant alpha-errors and thus do not sufficiently improve discrimination of "truly" activated pixels even for very high contrast-to-noise ratios (CNR). Only relative threshold strategies related to the maximum correlation coefficient and thus the individual data quality and activation level, i.e., a data-driven approach, can perfectly discriminate true positives, at least for CNR > 2.5. To further improve discrimination of activated and non-activated pixel in studies with lower CNR, additional prior knowledge would be necessary. From the data presented, one would also expect that the best performing threshold strategy in this simulation study would perform best under in vivo conditions.

Biometry↗

PubMed2001

[Real-Time Quantitative PCR and Detection of Minimal Residual Disease in Blood Malignancy]

Real-time quantitative PCR (RQ-PCR) is a recently developed technique for nuclear acid quantification, in which conventional PCR and probe hybridization are integrated together. A specific fluorescent-labeled probe, binded with DNA/cDNA target, is supplemented into the reaction system of PCR. During the extension phase of PCR amplification, the specific fluorescent-labeled probe is cleaved by the 5'-->3' exonuclease activity of Taq polymerase, producing a fluorescent signal that can be detected. The amount of fluorescence released in the reaction is proportional to the target sequence number. Thus, an exactly and absolutely quantitative assessment of initial target DNA/cDNA copy number can be made in a closed tube assay system. This technique eliminates the need for post-PCR sample processing which is necessary in conventional PCR and decreases the probability of contamination which is inevitable in conventional PCR. Moreover, it reduces the potiental for false positive or negative results by utilizing the hybridization of a probe to sequences internal to amplification primers. RQ-PCR is proved to be a sensitive, specific, reliable and convenient tool for the quantification of minimal residual diseases (MRD) of hematological malignancies by recent studies. This article reviews the principle of RQ-PCR and its application in quantitative detection of MRD of hematological malignancies such as acute myelogenous leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML) and malignant lymphoma.

Journal Article↗