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Calcification of coronary arteries: detection and quantification with double-helix CT.

PURPOSE: To evaluate the accuracy of double-helix computed tomography (CT) in coronary artery calcification detection and quantification. MATERIALS AND METHODS: One hundred sixty patients with coronary disease (135 men, 25 women; age range, 45-62 years), of whom 138 had obstructive (stenosis of > 50% of diameter; n = 129) or mild (< 50% stenosis; n = 9) coronary artery disease (CAD) and 22 had normal coronary arteries (per angiographic findings), and 56 age-matched healthy control subjects underwent double-helix CT. RESULTS: Double-helix CT findings indicated that calcification was significantly more prevalent in patients with CAD (> 83%) than in patients with normal coronary arteries (27%) or in healthy control subjects (34%; P < .01). Sensitivity in detecting obstructive CAD was high (91%); however, specificity was low (52%) because of calcification in nonobstructive lesions. Comparison of double-helix CT and angiographic findings indicated that double-helix CT was 84% accurate with positive and negative predictive values of 89% and 59%, respectively. CONCLUSION: Double-helix CT is a useful noninvasive method for detection and quantification of coronary artery calcification.

Adult↗

Pulmonary emphysema: subjective visual grading versus objective quantification with macroscopic morphometry and thin-section CT densitometry.

PURPOSE: To compare subjective visual grading of pulmonary emphysema with macroscopic morphometry and computed tomographic (CT) densitometry. MATERIALS AND METHODS: In 62 consecutive patients who underwent thinsection CT before surgical lung resection, emphysema was objectively quantified with computer-assisted macroscopic morphometry and CT densitometry. The percentage of lung macroscopically occupied by emphysema was compared with the percentage occupied on CT scans by pixels with attenuation values lower than a predefined threshold (CT densitometry). Three readers with varying degrees of expertise subjectively graded emphysema with visual assessment at two reading sessions. Data from objective quantification and subjective grading were analyzed with correlation coefficients, and interobserver and intraobserver agreement were calculated. RESULTS: Subjective grading of emphysema showed less agreement with the macroscopic reference standard results (r = 0.439-0.505; P < .05) than with objective CT densitometric results (r = 0.555-0.623; P < .001). The 95% CIs for the intercepts of the linear regression lines were suggestive of systematic subjective overestimation of emphysema by all three readers. Interobserver agreement was moderate (kappa = 0.431-0.589). Intraobserver agreement was good to excellent (kappa = 0.738-0.936). The expertise of individual readers did not substantially influence results. CONCLUSION: Systematic overestimation and moderate interobserver agreement may compromise subjective visual grading of emphysema, which suggests that subjective visual grading should be supplemented with objective methods to achieve precise, reader-independent quantification of emphysema.

Absorptiometry, Photon↗

Quantification of viral load: clinical relevance for human immunodeficiency virus, hepatitis B virus and hepatitis C virus infection.

Quantitative determination of viral load using nucleic acid amplification techniques represents the most accurate prognostic marker for human immunodeficiency virus type 1 (HIV-1) infection, independently of CD4+ cell count. Overall, the different methods for HIV-1 RNA determination (RT-PCR, nucleic acid sequence-based amplification, branched DNA) show a good reproducibility (0.5 log), however for low copy numbers and in HIV-1-infected children the variability may exceed 0.7 log. In non-HIV-1 subtype B infections the copy number is underestimated. While serology permits an accurate follow-up of hepatitis B virus (HBV) infection, HBV DNA quantification is used for monitoring of antiviral therapy, determination of infectiosity and in combination with serological markers for the resolution of unusual profiles, i.e. isolated anti-HBc reactivity. The prognostic relevance of hepatitis C virus (HCV) RNA determination is of limited value for the long-term prognosis of chronic hepatitis C, however the viral load may predict the outcome of antiviral therapy. Genetic diversity represents a challenge for HCV RNA quantification.

Acquired Immunodeficiency Syndrome↗

Quantification of osteoclastic resorption of the bovine otic capsule in vitro by an enzyme-linked immunosorbent assay.

The bony shell surrounding the inner ear is known to have a very pronounced centripetal inhibition of remodelling in vivo, with almost no bone turnover immediately adjacent to the perilymphatic spaces and a gradually increasing turnover rate towards outer parts of the bony otic capsule. By the use of in vitro markers of bone resorption, including an enzyme-linked immunosorbent assay for quantification of type I collagen degradation and a colorimetric enzyme assay for quantification of osteoclast tartrate-resistant acid phosphatase activity, this study demonstrates that there are no ex vivo differences in bone matrix resorption between the inner and outer parts of the otic capsule when exposed to seeded osteoclasts from rabbits. Thus, the unique spatial distribution of perilabyrinthine bone turnover is not caused by a shift in resorbability from inner to outer capsular bone that is due to inherent bone quality differences particular to these bone compartments. More likely, the sustained action of some intravital 'field force', originating from the inner ear spaces, is responsible for the unique spatial distribution of the otic capsular bone turnover found in vivo. Though the character of this force is not yet defined, it is appealing to relate it to the large electromagnetic potential gradient present in the inner ear.

Analysis of Variance↗

Comparison of QLF and DIAGNOdent for quantification of smooth surface caries.

The aim was to compare the detection and quantification of smooth surface caries by KaVo DIAGNOdent and quantitative laser/light-induced fluorescence (QLF) using a laser and a lamp as two different light sources. On 40 premolar teeth extracted from adolescents, 71 non-cavitated approximal surfaces were assessed. For QLF, both mean and maximum fluorescence losses were registered. To provide a gold standard for verification, the teeth were then sectioned and lesion depth, stratified on a five-point scale, was determined by histopathology and microradiography. The correlation between the gold standard and the two methods was assessed by Spearman's rank correlation coefficient. The sensitivity and specificity of the methods for detection of smooth surface caries at D(3) level were also assessed. For lesion depth, correlation with the gold standard was similar for QLF and DIAGNOdent: about 0.85. With respect to dentinal caries detection, sensitivity for DIAGNOdent was 0.75 and specificity 0.96, with a cut-off point of 9. The corresponding values for QLF were 0.94 and 1, with a cut-off value of 20% of fluorescence loss. Spearman's rank correlation coefficients for enamel mineral loss and QLF and DIAGNOdent, respectively, were 0.76 and 0.67. It was concluded that for quantification of smooth surface caries, the methods are of equal merit, but for scientific purposes, QLF offers the advantage of closer correlation with changes in mineral content.

Adolescent↗

Quantification of pancreatic zinc output as pancreatic function test: making the secretin-caerulein test applicable to clinical practice.

BACKGROUND/AIM: The secretin-caerulein test (SCT) is generally considered the gold standard for evaluation of the exocrine pancreatic function. Problems related to enzyme inactivation in the aspirated duodenal juice limit the clinical applicability of the test. Pancreatic zinc, which is mainly secreted as constituent of metalloenzymes, is stable in duodenal juice and easy to quantify. The aim of this study was to analyze the accuracy of the secretin-caerulein-stimulated pancreatic zinc output as pancreatic function test in comparison with the standard SCT. METHODS: Forty consecutive patients with suspected chronic pancreatitis and 28 healthy subjects were studied. A SCT was performed after overnight fast by infusing intravenously secretin (1 U/kg/h) and caerulein (100 ng/kg/h) over 90 min. The duodenal content was continuously aspirated and separated at 15-min intervals and immediately analyzed for pH, bicarbonate, amylase, lipase, elastase, carboxypeptidase A, and zinc. Correlation and concordance between standard SCT and quantification of zinc output and the accuracy of the latter for diagnosing and grading the exocrine pancreatic dysfunction were calculated. RESULTS: The pancreatic zinc output correlated significantly with enzyme and bicarbonate output (r ranging from 0.670 to 0.855; p < 0.001). A highly significant concordance was found between the degree of exocrine pancreatic dysfunction based on the standard SCT (bicarbonate and enzymes output) and that based only on zinc output (k = 0.831; p < 0.001). Quantification of the stimulated pancreatic zinc output has a sensitivity of 97% and a specificity of 91% in the diagnosis of exocrine pancreatic dysfunction. CONCLUSIONS: The determination of pancreatic zinc output during secretin and caerulein stimulation is a simple and accurate method for evaluation of the exocrine pancreatic function. Zinc is stable in duodenal juice, and its determination as a single parameter simplifies the clinical applicability of the SCT.

Adolescent↗

A new method of color Doppler perfusion measurement via dynamic sonographic signal quantification in renal parenchyma.

OBJECTIVE: Perfusion quantification of tissues is an important goal to evaluate the state of blood supply of an organ. We developed a method to quantify tissue perfusion via color Doppler signal quantification from sonographic videos and applied this to describe renal parenchymal perfusion in healthy kidneys. METHOD: Color Doppler sonographic videos of renal perfusion from both kidneys of 87 healthy children (age 2 weeks to 16 years) were recorded under defined conditions. Perfusion data (color hue, color area) were measured in a standardized region of interest automatically. Signal intensity was calculated as whole ROIs (regions of interest) mean flow velocity (cm/s) encoded by color Doppler signals during one full heart cycle. RESULTS: Normal signal intensity values are: 1.86 cm/s in the region encompassing central 50% of the renal cortex and 0.56 cm/s in the peripheral 50% of the renal cortex. These differences are significant. Signal intensity of both kidneys did not differ. CONCLUSION: Signal intensity of cortical tissue in healthy kidneys was quantified noninvasively from color Doppler signal data in an easily accomplishable manner with new measurement software. Normal values for this technique have been calculated. Possible further applications might be all situations, where perfusion changes could be expected like inflammation, renal insufficiency, vascular diseases and tumors.

Adolescent↗

Quantification of the major allergen from cypress (Cupressus arizonica) pollen, Cup a 1, by monoclonal antibody-based ELISA.

BACKGROUND: Cypress pollen allergy is an important cause of rhinoconjunctivitis and asthma in Mediterranean countries. Cypress allergenic extracts are difficult to produce since they have low protein and high carbohydrate content, thus accurate standardization of them is essential to guarantee their quality. The aim of this study is to develop a sandwich ELISA for the quantification of Cup a 1, the major allergen of cypress (Cupressus arizonica) pollen extract. METHODS: Monoclonal antibodies directed to purified Cup a 1 were produced. Two of them (9C7 as capture antibody and 3D2 as the tracer) were selected to develop a quantitative sandwich ELISA. This ELISA was subsequently evaluated and compared with other techniques. RESULTS: The described ELISA is very sensitive with a detection limit of 8.7 ng/ml and a practical working range of 62.5-1,000 ng/ml. The assay is also highly reproducible with intra-assay and interassay coefficients of variation of less than 10%. The purified Cup a 1, used as standard, presents pectate lyase enzymatic activity. The assay also detected Cup a 1-like proteins in pollen from other Cupressaceae. A good correlation was obtained between Cup a 1 content of 12 C. arizonica pollen extracts and their IgE-binding activity. CONCLUSIONS: The described Cup a 1 ELISA is sensitive, specific and reproducible and can be used for the quantification of Cup a 1 in C. arizonica and other related pollen extracts. It also provides a reliable indication of the allergenic activity of the whole cypress pollen extract.

Allergens↗

Interest of fluorine-19 nuclear magnetic resonance spectroscopy in the detection, identification and quantification of metabolites of anticancer and antifungal fluoropyrimidine drugs in human biofluids.

BACKGROUND: The metabolism of fluorouracil and fluorocytosine, two 5-fluoropyrimidine drugs in clinical use, was investigated. METHODS: (19)F nuclear magnetic resonance (NMR) spectroscopy was used as an analytical technique for the detection, identification and quantification of fluorinated metabolites of these drugs in intact human biofluids as well as fluorinated degradation compounds of fluorouracil in commercial vials. RESULTS: (19)F NMR provides a highly specific tool for the detection and absolute quantification, in a single run, of all the fluorinated species, including unexpected substances, present in biofluids of patients treated with fluorouracil or fluorocytosine. Besides the parent drug and the already known fluorinated metabolites, nine new metabolites were identified for the first time with (19)F NMR in human biofluids. Six of them can only be observed with this technique: fluoride ion, N-carboxy-alpha-fluoro-beta-alanine, alpha-fluoro-beta-alanine conjugate with deoxycholic acid, 2-fluoro-3-hydroxypropanoic acid, fluoroacetic acid, O(2)-beta-glucuronide of fluorocytosine. CONCLUSION: (19)F NMR studies of biological fluids of patients treated with anticancer fluorouracil or antifungal fluorocytosine have furthered the understanding of their catabolic pathways.

Antifungal Agents↗

Agreement between the classical urea kinetic model and direct dialysis quantification: importance of urea rebound.

From a review of the literature regarding kinetic models used for assessing the adequacy of hemodialysis, no definite conclusions can be drawn as to whether the classical urea kinetic model (UKM) or modified direct dialysis quantification (mDDQ) is more reliable. We compared mDDQ with classical UKM and with a modified UKM that employs an equilibrated urea value. From the theoretical viewpoint, no substantial conflict is found between the two models as regards the dialysis dose, if urea rebound is considered. From the practical viewpoint, in our opinion direct quantification lends itself better for experimental purposes whereas for routine Kt/V evaluation UKM is easier and accurate enough, provided that rebound is taken into account.

Female↗

Quantification of fibronectin in bronchoalveolar lavage fluid and in lung parenchyma: differences between smokers and non-smokers.

Fibronectin (FN) is a glycoprotein present in the epithelial surface of airways and in the interstitial parenchyma of the lung. The aim of this study was to quantify the amount of FN in human lung parenchyma, and to investigate possible differences between smokers and non-smokers, both in lung parenchyma and in bronchoalveolar lavage (BAL) fluid. BAL fluid was recovered in 21 individuals. Quantification of FN in BAL was done by a non-competitive ELISA. Twenty-six lung parenchyma samples were also obtained from lung resections. FN was extracted with urea and heparin. Quantification of FN in lung parenchyma was determined by a competitive ELISA. The mean +/- SD FN content in BAL fluid from smokers was 1.59 +/- 1.65 micrograms/mg total protein and 0.53 +/- 0.26 in non-smokers (p < 0.05). However, the amount of FN/ml BAL fluid was not statistically different between smokers (0.15 +/- 0.13 micrograms/ml) and non-smokers (0.106 +/- 0.07). The mean extractable FN in human lung tissue was 38.6 micrograms/g wet weight (31.7-45.5, 95% mean confidence interval) with no statistical difference between smokers and non-smokers. We conclude that in smokers, FN accounts for a higher proportion of the total protein in BAL fluid, and that there were no differences in FN content in lung parenchyma between smokers and non-smokers.

Bronchoalveolar Lavage Fluid↗

Quantification of angioscotomas.

The current normal visual-field indices to quantify scotomas are not applicable to the stimuli in angioscotometry. Therefore, different alternative methods for the quantification of angioscotomas are presented and discussed. The proposed method for quantification of angioscotomas can easily be calculated with commercially available statistical programs by every physician interested in statistics. A program for angioscotometry including an index calculation is proposed. Three examples illustrate the possible alternative calculations.

Adult↗

Detection and quantification of mature circulating endothelial cells using flow cytometry and immunomagnetic beads: a methodological comparison.

Mature circulating endothelial cells (CECs) are novel cellular markers of endothelial damage/dysfunction. The two main techniques of CEC enumeration are flow cytometry (FC) and immunomagnetic bead (IB) isolation. Both quantify CECs accurately, but a direct comparison of both methods has not been reported. We sought to assess the agreement between the two methods in two patient populations, and a group of healthy subjects, with emphasis given to methodological issues. We included 34 patients with acute coronary syndrome (ACS), 60 patients with primary breast cancer (PBC) and 30 healthy controls (HC). We quantified CECs using the IB method [CD146 and FITCUlex europaeus lectin-1] and FC [CD45, CD34 and CD146]. Bland-Altman plots suggested reasonable agreement (<5% of events >2 standard deviations from the mean) between FC and the IB methods for CEC quantification in whole blood in the two disease groups (ACS and PBC), but not among the HCs. There were no statistically significant differences in CEC levels by the two methods amongst all three patient groups. There is reasonable agreement between the FC and the IB methods for mature CEC quantification in whole blood, especially amongst disease groups. The agreement between the two methods appears to weaken in healthy controls, and at lower and higher absolute CEC counts.

Adult↗

Noninvasive quantification of atherosclerotic lesions. Reproducibility of ultrasonographic measurement of arterial wall thickness and plaque size.

A noninvasive method based on high-resolution B-mode ultrasonography and a computerized image-analyzing system were used for the quantification of early (thickening of the intima-media complex) and late (plaque) atherosclerosis in the carotid and the femoral artery. The difference between repeated measurements was assessed to estimate intraobserver and interobserver differences. The results were satisfactory, with a coefficient of variation for measurement of lumen diameter in the common carotid artery of 3.8 +/- 4.1% (r = 0.91) and for the femoral artery of 4.8 +/- 4.1% (r = 0.93). Corresponding figures for intima-media thickness in the common carotid artery and the femoral artery were 10.0 +/- 8.5% (r = 0.86) and 16.2 +/- 12.6% (r = 0.91), respectively. The coefficient of variation for measurements of maximal intima-media thickness at the site of the plaque was 14.6 +/- 10.5% (r = 0.88); for plaque base, 13.1 +/- 9.0% (r = 0.91); and for plaque area, 16.3 +/- 12.6% (r = 0.90). The method seems promising for the detection and quantification of early and late atherosclerotic lesions in the carotid and femoral arteries.

Aged↗

Quantification of cholesteryl esters in human and rabbit atherosclerotic plaques by magic-angle spinning (13)C-NMR.

Accumulation of cholesteryl esters (CEs) is a key event in the formation of atherosclerotic plaques. More recent work suggests a role for CEs in plaque rupture leading to thrombosis, which can result in an acute event such as myocardial infarction or stroke. In this study, we present nuclear magnetic resonance (NMR) protocols for quantification of CEs in plaques in situ. Total CEs quantified by (13)C magic-angle spinning (MAS) NMR in excised plaques from human carotid arteries and rabbit aortic arteries were in good agreement with the amounts determined by subsequent standard chemical assays. The latter analysis is disadvantageous because it requires that plaque lipids be extracted from the tissue, resulting in the loss of all phase information of CEs as well as other major plaque components. With our MAS-NMR protocol, the plaque components are preserved in their native phases. Combining MAS and off-MAS NMR, we were able to quantitatively distinguish isotropic (liquid) CEs from anisotropic (liquid-crystalline) CEs in plaque tissues. In a recent study, we applied a different (13)C MAS-NMR protocol to quantify crystalline cholesterol monohydrate in plaques. Together, these 2 studies describe a new, noninvasive MAS-NMR strategy for the identification and quantification of the major lipid components in plaques in situ. This approach will be useful for investigation of the relationship between plaque rupture and specific lipids in their biologically relevant phases.

Animals↗

Rapid left-to-right shunt quantification in children by phase-contrast magnetic resonance imaging combined with sensitivity encoding (SENSE).

BACKGROUND: Parallel imaging by sensitivity encoding (SENSE) may considerably reduce scan time in MRI. For rapid flow quantification in children with congenital heart disease, we evaluated phase-contrast MRI (PC-MRI) techniques combined with SENSE. METHODS AND RESULTS: In 22 pediatric patients (mean age, 7.2+/-6.2 years) with cardiac left-to-right shunt, blood flow rate in the pulmonary artery (Qp) and ascending aorta (Qs) and flow ratio Qp/Qs were determined by PC-MRI with SENSE reduction-factor 2 and 3 (SF-2 and SF-3). Additionally, we used PC-MRI with higher spatial in-plane resolution (1.6x2.1 versus 2.3x3.1 mm) with and without SF-3. Results were compared with a recently validated standard PC-MRI protocol and tested in vitro using a pulsatile flow phantom. Reduction of signal averages from 2 to 1 and application of SENSE accelerated flow measurements by a factor of 3.5 (5.2) using PC-MRI with SF-2 (SF-3) compared with standard PC-MRI. For blood flow rate through the pulmonary artery and aorta, as well as for the Qp/Qs ratio we found negligible differences of +/-3%, lower limits of agreement (mean+/-2 SD) of -7% to -18%, and upper limits of agreement (mean+/-2 SD) of +3 to +24%, demonstrating good agreement with standard PC-MRI. Mean Qp/Qs ratio by standard PC-MRI was 1.69+/-0.45 (range, 1.27 to 2.79). Interobserver variability was low, and high accuracy was confirmed in vitro for all protocols. CONCLUSIONS: PC-MRI for flow quantitation may be combined with SENSE to achieve a substantive reduction of scanning time. In children with left-to-right shunt, Qp/Qs quantification is possible by PC-MRI+SF-3 in <60 seconds. Use of higher in-plane resolution did not improve measurement results.

Adolescent↗

Flow volume and shunt quantification in pediatric congenital heart disease by real-time magnetic resonance velocity mapping: a validation study.

BACKGROUND: Flow quantification in real time by phase-contrast MRI (PC-MRI) may provide unique hemodynamic information in congenital heart disease, but available techniques have important limitations. We sought to validate a novel real-time magnetic resonance flow sequence in children. METHODS AND RESULTS: In 14 pediatric patients (mean age 5.2+/-2.0 years) with cardiac left-to-right shunt, pulmonary (Q(p)) and aortic (Q(s)) flow rates were determined by nontriggered free-breathing real-time PC-MRI with single-shot echo-planar imaging combined with sensitivity encoding, which yielded 25 phase images per second at 2.7x2.7-mm in-plane resolution (field of view 30x34 cm2). Over a 9.5-second period that included 2 to 5 respiratory cycles, 16.6+/-2.6 subsequent stroke volumes (range 13 to 22) were acquired in each vessel. Results were compared with conventional retrospectively ECG-gated PC-MRI. Mean Q(p)/Q(s) by conventional PC-MRI was 1.91+/-0.64, and it was 1.94+/-0.68 (mean+/-SD) by real-time PC-MRI. For blood flow rate through pulmonary artery and aorta, we found differences of 2% to 3% (Bland-Altman analysis), with lower limits of agreement of -11% to -13% (mean-2 SD) and upper limits of 18% to 19% (mean+2 SD), which demonstrated good agreement between both methods. Mean difference for Q(p)/Q(s) was 1%, with limits of agreement ranging between -18% and 22% (mean+/-2 SD). High repeatability but some flow overestimation was observed in vitro (pulsatile flow phantom) with real-time PC-MRI, whereas conventional PC-MRI was accurate. Beat-to-beat stroke-volume variation was 6.1+/-2.3% in vivo and 3.7+/-0.3% in vitro. CONCLUSIONS: Beat-to-beat quantification of pulmonary and aortic flows and hence left-to-right shunt within a few seconds is reliable by nontriggered real-time PC-MRI with echo-planar imaging and sensitivity encoding. Good spatial/temporal resolution and a large field of view may render the sequence valuable for multiple applications in congenital heart disease.

Blood Circulation↗

Noninvasive detection and quantification of left-to-right shunts in children using oxygen-15 labeled carbon dioxide.

A method for the detection and quantification of left-to-right intracardiac shunts is described which utilizes a single breath inhalation of oxygen-15 labeled carbon dioxide (C15O2). The inhaled gas rapidly crosses the alveolar membrane and the oxygen-15 label is exchanged through the carbonate cycle to form oxygen-15 labeled water within the pulmonary capillary blood. Pulmonary indicator clearance curves are measured by external scintillation probes. A simplified method of shunt flow quantification was developed from indicator dilution principles and used for the analysis of the clearance curves. Inhalation studies were performed with 62 children on the day prior to cardiac catheterization. The presence or absence of left-to-right shunt was confirmed by contrast angiography in all cases. Twenty-six children were found to have no shunts by C15O2 inhalation, oximetry or angiography. Of the 36 with shunts, 34 were detected by C15O2. Two of these were designated as equivocal because they were considered to be less than the threshold of definitive detection by C15O2 (having Qp/Qs less than 1.2); 32 were positive and there were two false negatives with small ventricular septal defects. There were no false positives by C15O2. The correlation coefficient between C15O2 and oximetry values of shunt flow for those patients with proven shunts was 0.82.

Adolescent↗