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At least 1,153 records · Page 64Linked to original sources
PubMed1998

Quantification of mitral regurgitation by automated cardiac output measurement: experimental and clinical validation.

OBJECTIVES: To develop and validate an automated noninvasive method to quantify mitral regurgitation. BACKGROUND: Automated cardiac output measurement (ACM), which integrates digital color Doppler velocities in space and in time, has been validated for the left ventricular (LV) outflow tract but has not been tested for the LV inflow tract or to assess mitral regurgitation (MR). METHODS: First, to validate ACM against a gold standard (ultrasonic flow meter), 8 dogs were studied at 40 different stages of cardiac output (CO). Second, to compare ACM to the LV outflow (ACMa) and inflow (ACMm) tracts, 50 normal volunteers without MR or aortic regurgitation (44+/-5 years, 31 male) were studied. Third, to compare ACM with the standard pulsed Doppler-two-dimensional echocardiographic (PD-2D) method for quantification of MR, 51 patients (61+/-14 years, 30 male) with MR were studied. RESULTS: In the canine studies, CO by ACM (1.32+/-0.3 liter/min, y) and flow meter (1.35+/-0.3 liter/min, x) showed good correlation (r=0.95, y=0.89x+0.11) and agreement (deltaCO(y-x)=0.03+/-0.08 [mean+/-SD] liter/min). In the normal subjects, CO measured by ACMm agreed with CO by ACMa (r=0.90, p < 0.0001, deltaCO=-0.09+/-0.42 liter/min), PD (r=0.87, p < 0.0001, deltaCO=0.12+/-0.49 liter/min) and 2D (r=0.84, p < 0.0001, deltaCO=-0.16+/-0.48 liter/min). In the patients, mitral regurgitant volume (MRV) by ACMm-ACMa agreed with PD-2D (r= 0.88, y=0.88x+6.6, p < 0.0001, deltaMRV=2.68+/-9.7 ml). CONCLUSIONS: We determined that ACM is a feasible new method for quantifying LV outflow and inflow volume to measure MRV and that ACM automatically performs calculations that are equivalent to more time-consuming Doppler and 2D measurements. Additionally, ACM should improve MR quantification in routine clinical practice.

Adult↗
PubMed2002

Real-time RT-PCR quantification of insulin-like growth factor (IGF)-1, IGF-1 receptor, IGF-2, IGF-2 receptor, insulin receptor, growth hormone receptor, IGF-binding proteins 1, 2 and 3 in the bovine species.

Reverse transcription (RT) followed by polymerase chain reaction (PCR) is the technique of choice for analysing mRNA in extremely low abundance. Real-time RT-PCR using SYBR Green I detection combines the ease and necessary exactness to be able to produce reliable as well as rapid results. To obtain highly accurate and reliable results in a real-time RT-PCR a highly defined calibration curve is needed. We designed and developed nine different calibration curves, based on recombinant DNA plasmid standards and established them on a constant real-time PCR platform for the following factors: growth hormone receptor (GHR), insulin-like growth factor (IGF)-1, IGF-1 receptor (IGF-1R), IGF-2, IGF-2 receptor (IGF-2R), insulin receptor (INSR), and IGF-binding proteins (IGF-BP) 1, 2 and 3. Developed assays were applied in the LightCycler system on bovine ileum and liver total RNA and showed high specificity and sensitivity of quantification. All assays had a detection limit of under 35 recombinant DNA molecules present in the capillary. The SYBR Green I determination resulted in a reliable and accurate quantification with high test linearity (Pearson correlation coefficient r > 0.99) over seven orders of magnitude from <10(2) to >10(8) recombinant DNA start molecules and an assay variation of maximal 5.3%. Applicability of the method was shown by analysing mRNA levels in newborn calves: mRNA concentrations per gram tissue of mRNAs of IGF-1, IGF-1R, IGF-2, IGF-2R, GHR, INSR, and IGF-BP1, 2 and 3 were all different between in liver and ileum and the traits all exhibited individual differences.

Animals↗
PubMed2000

Detailed methods for the quantification of nitric oxide in aqueous solutions using either an oxygen monitor or EPR.

The interest in nitric oxide has grown with the discovery that it has many biological functions. This has heightened the need for methods to quantify nitric oxide. Here we report two separate methods for the quantification of aqueous stock solutions of nitric oxide. The first is a new method based on the reaction of nitric oxide with oxygen in liquid phase (*NO + O2 + 2H2O --> 4HNO2); an oxygen monitor is used to measure the consumption of oxygen by nitric oxide. This method offers the advantages of being both simple and direct. The presence of nitrite or nitrate, frequent contaminants in nitric oxide stock solutions, does not interfere with the quantification of nitric oxide. Measuring the disappearance of dissolved oxygen, a reactant, in the presence of known amounts of nitric oxide has provided verification of the 4:1 stoichiometry of the reaction. The second method uses electron paramagnetic resonance spectroscopy (EPR) and the nitric oxide trap [Fe2+-(MGD)2], (MGD = N-methyl-D-glucamine dithiocarbamate). The nitrosyl complex is stable and easily quantitated as a room temperature aqueous solution. These two methods are validated with Sievers 280 Nitric Oxide Analyzer and cross-checked with standards using UV-Vis spectroscopy. The practical lower limits for measuring the concentration of nitric oxide using the oxygen monitor approach and EPR are approximately 3 microM and 500 nM, respectively. Both methods provide straightforward approaches for the standardization of nitric oxide in solution.

Electron Spin Resonance Spectroscopy↗
PubMed1987

Quantification of 5- and 15-HPETE's by direct chemical ionization mass spectrometry.

This report describes the application of direct chemical ionization mass spectrometry (DCIMS) to the identification and quantification of 5- and 15-HPETEs. A unique feature of the method is use of a polyimide-coated fused silica fiber that allows vaporization of the hydroperoxides, with very low excess energy, into the plume of the chemical ionization reagent gas plasma. Mass spectra are obtained that allow identification of the nonreduced and nonderivatized free acid forms of 5- and 15-HPETE as well as their quantification from 1 microgram to 100 picograms.

Animals↗
PubMed1998

ESR and HPLC-EC analysis of ethanol oxidation to 1-hydroxyethyl radical: rapid reduction and quantification of POBN and PBN nitroxides.

Extensive ESR spin-trapping studies have shown that ethanol is oxidized to 1-hydroxyethyl radical (HER) by rat and deer mice liver microsomal systems. The ESR detection of POBN/HER nitroxide in bile, and formation of antibodies, which recognize HER adducts in alcoholics, suggest that HER is produced in vivo. In liver, where ethanol is primarily metabolized, only traces of PBN/HER nitroxide are documented. One limitation of the ESR spin-trapping technique, however, is that the nitroxides formed in the presence of cellular reductants can be metabolized to the corresponding ESR "silent" hydroxylamines. Ascorbate and NADPH plus liver microsomes were found to reduce rapidly both POBN/HER and PBN/HER nitroxides to their ESR "silent" hydroxylamine derivatives. An HPLC method with electrochemical detection was developed for the detection and quantification of both POBN/HER and PBN/HER nitroxides, as well as their hydroxylamines. Both the diastereomers of the POBN/HER nitroxide and hydroxylamine can be detected, as can both isomers of the PBN/HER nitroxide, and it is estimated that the sensitivity of the HPLC procedure is in the nM range when using EC detection. The hydroxylamines are stable in ethanol, while pH-dependent auto-oxidation occurs in aqueous buffers. Some of the characteristics associated with HER formation by microsomes as detected with ESR (e.g., sensitivity to SOD and catalase, increase after induction of CYP2E1) are reproduced with the HPLC method. By quantification of the POBN/HER hydroxylamine, the NADPH-dependent rates of HER formation by microsomes from pyrazole-treated rats are estimated to be about 1-1.5 nmol HER per min per mg microsomal protein. This rate is less, as compared to the two electron-dependent rate of acetaldehyde formation by these microsomes, about 10-15 nmol per min per mg protein. Thus, at first approximation, the one electron-dependent rate of ethanol oxidation is about 10% the two electron-dependent rate by isolated microsomes. The HPLC procedure can readily detect the POBN/HER and PBN/HER nitroxides and their hydroxylamine derivatives in the same sample and may be of value in detecting HER spin-trapped adducts under biological reducing conditions.

Animals↗
PubMed1998

Quantification of mitral regurgitation by the automated cardiac output method: an in vitro and in vivo study.

BACKGROUND: Recently, the automated cardiac output method (ACM) was introduced for the calculation of blood flow at the left ventricular outflow tract (LVOT). This study was performed to examine the possibility of using ACM for flow calculation at the level of the mitral valve and for the quantification of mitral regurgitation (MR) in vitro and in vivo. METHODS AND RESULTS: In a computer-controlled in vitro model of the human heart, aortic and mitral normal bioprosthetic valves were inserted. ACM and electromagnetic probe flow measurements correlated well at the LVOT and at the mitral level (r2 = 0.79 and 0.77, respectively). For stroke volumes ranging from 30 to 100 ml/beat, there was no statistically significant bias between ACM and electromagnetic flow probe (-1.5 and 1.3 ml for LVOT and mitral level, respectively). Limits of agreement were [-14; +11] ml and [-18; +16] ml, respectively. We evaluated 68 patients in our in vivo study. They were divided into three groups according to the results of "standard" echocardiographic Doppler methods for the semiquantification of MR: echocardiographic color Doppler cartography, intensity of the continuous wave Doppler spectra, and in some patients, pulmonary venous flow, conventional Doppler, and proximal isovelocity surface area quantitative data. Group 1 consisted of 35 patients without MR or a physiologic one; the 17 patients in group 2 had a mild MR (1-2/4) and in group 3, 16 patients with MR 3-4/4 were included. Regurgitant volume (RV) was calculated as the difference between ACM mitral flow and ACM aortic flow, and regurgitant fraction (RF) was defined as the ratio between RV and ACM mitral flow. When mitral flow was measured only from the four-chamber view, we found in group 1, RV = -0.57 (0.67) L/min and RF = -16% (19%); in group 2, RV = -0.31 (1.06) L/min and RF = -8% (19%); and in group 3, RV = 1.53 (0.94) L/min and RF = 23% (13%). RV and RF were statistically higher in group 3 compared with group 2 or group 1 (p < 0.0005), but no significant difference was found between groups 1 and 2. When mitral flow was measured by the mean value of ACM four-chamber and two-chamber views, this resulted in group 1, RV = -0.26 (0.63) L/min and RF = -8% (15%); in group 2, RV = 0.01 (1.04) L/min and RF = -2% (18%); and in group 3, RV = 2.07 (1.21) L/min and RF = 34% (19%). RV and RF were again significantly higher in group 3 (p < 0.0001). There was no significant difference between group 1 and group 2, but in group 1 RF was no longer statistically different from 0%. CONCLUSIONS: (1) In our in vitro setting, ACM is reliable both at the LVOT and at the mitral valve. (2) In the in vivo situation, some overlapping does exist between the three groups of MR. However, ACM is a very easy, rapid, and objective method to differentiate hemodynamic nonsignificant (<3/4) from significant (> or =3/4) MR. Together with other well-known methods for the quantification of MR, it should facilitate the gradation of MR in the clinical setting. The absence of significant differences between group 1 and group 2 proves that the accuracy of ACM measurements at the mitral valve needs to be ameliorated before ACM can be used as a gold standard for the noninvasive measurement of RV and RF.

Aortic Valve↗
PubMed1998

Acoustic quantification indexes of left ventricular size and function: effects of signal averaging.

BACKGROUND: The aim of this study was to evaluate the clinical utility of using signal-averaged acoustic quantification (SAAQ) waveforms for improved assessment of left ventricular (LV) size and function. METHODS AND RESULTS: Four separate protocols were performed in 47 subjects. SAAQ waveforms were used to assess alterations in LV function induced by dobutamine (15 microg/kg per minute) and esmolol (200 microg/kg per minute) in eight normal subjects. Subsequently, we analyzed SAAQ waveforms obtained in 12 patients with LV dysfunction secondary to dilated cardiomyopathy and 12 age-matched normal subjects. Finally, we developed computer software for monitoring of LV function on the basis of continuous acquisition and repeated analysis of SAAQ waveforms. We compared the interbeat variability in indexes of LV function obtained from raw AQ and SAAQ during 10 minutes of steady-state monitoring in eight patients undergoing transesophageal echocardiography. The feasibility of long-term monitoring in the intensive care setting was then studied in seven patients undergoing abdominal surgery. Our analysis tracked variations in LV function induced by dobutamine and esmolol. Significant differences in all measured indexes were found between normal subjects and patients with dilated cadiomyopathy. Signal averaging during steady-state monitoring significantly reduced the interbeat variability of all indexes (21% to 42%). In the operating room, the SAAQ monitoring system tracked hemodynamic changes in close agreement with invasive measurements. CONCLUSIONS: SAAQ allows fast and easy quantification of LV function and can track hemodynamic trends in the operating room setting.

Adrenergic beta-Agonists↗
PubMedDate not supplied

Detection and quantification of the iap gene of Listeria monocytogenes and Listeria innocua by a new real-time quantitative PCR assay.

A real-time quantitative polymerase chain reaction (PCR) assay for direct detection and enumeration of Listeria monocytogenes and Listeria innocua was developed and applied to artificially contaminated milk samples. The iap gene present in both species was used as a target for amplification of a 175-bp (L. monocytogenes) and a 309-bp (L. innocua) fragment. To ensure that L. monocytogenes and L. innocua are specifically detectable, tests were carried out using 42 L. monocytogenes strains and 33 L. innocua strains belonging to different serovars. Specificity was also confirmed using 22 bacterial strains not belonging to the genus Listeria, including closely related bacteria. In addition to specificity, the reported assay is characterized by a wide dynamic range of quantification and a high sensitivity, as we could detect as few as six copies of the iap gene per PCR using purified DNA as template. When applied to direct detection and quantification of L. monocytogenes in milk, the more rapid real-time quantitative PCR assay was as sensitive as the traditional plate count method, but real-time quantitative PCR-derived iap gene copy numbers were one to two logs higher than colony-forming units obtained by the plate count method.

Animals↗
PubMed2000

Methods for ELNES-quantification: characterization of the degree of inversion of Mg-Al-spinels

The energy loss near-edge structures in electron energy-loss spectra contain information about bonding characteristics, the electronic structure and coordinations of the excited atoms. We have calculated sets of reference spectra for the normal and for the inverse Mg-Al-spinel using a full multiple scattering approach. By a quantitative comparison of these reference spectra with experimental data ELNES-quantification becomes possible. We characterized the degree of inversion lambda by the analysis of the relative peak-intensities and the relative peak-positions within 35 eV beyond the edge onset. The results demonstrate that by using the provided methods ELNES-quantification will become possible when uncertainties in the experiment are reduced and a better fit of the simulations to the experiment is achieved.

Journal Article↗
PubMed1999

Quantification aspects of patient studies with 52Fe in positron emission tomography.

Quantification accuracy in positron emission tomography (PET) using non-pure positron emitters, such as 52Fe, may be influenced by gamma radiation emitted in the decay of these isotopes. High-energy positrons, emitted in the decay of the 52Fe-daughter 52mMn, also affect the quantification accuracy. A specific problem of the 52Fe/52mMn decay chain in vivo is that the kinetics of iron and manganese are different, and that PET cannot discriminate between the two nuclides. The effect of the decay properties of 52Fe/52mMn on the performance of PET was investigated using phantoms. Minor degradation in PET performance was found for 52Fe/52mMn compared to the pure low-energy positron emitter 18F. A method is presented to obtain a correction factor for the 52mMn radioactivity in blood. A model for correction of 52mMn-radioactivity in organs, based on existing data on manganese kinetics, is given. The presented corrections are discussed and illustrated in a patient study.

Adult↗
PubMed2003

Quantification of brain mu-opioid receptors with [11C]carfentanil: reference-tissue methods.

[(11)C]Carfentanil (CFN) is a mu-opioid agonist used for in vivo positron emission tomography (PET) studies of mu-opioid receptors. Previously, a tissue-ratio method was validated for the quantification of CFN binding. However, since that initial validation, several other blood independent (reference-tissue) methods have become available. To evaluate these methods, CFN PET studies with arterial blood sampling were acquired in six healthy male control subjects. Specific binding estimates obtained from reference-tissue methods were compared to those obtained with a more rigorous blood input modeling technique. It was determined that both a graphical method, and a simplified reference tissue model, were more accurate than the tissue-ratio method for quantification of CFN binding.

Adult↗
PubMed2000

Utilization of MS3 spectra for the multicomponent quantification of diastereomeric N-acetylhexosamines.

A rapid and accurate means of quantifying mixtures of diastereomeric N-acetylhexosamine monosaccharides using MS3 product ions is introduced. The method involves derivatizing the monosaccharides with [Co(DAP)2Cl2]Cl (where DAP is diaminopropane), and subjecting the derivatized products to collision-induced dissociation (CID) in a quadrupole ion trap mass spectrometer. Each diastereomer provides unique MS3 product ion abundances. The abundances for the pure monosaccharide standards are used in a system of equations in order to quantify mixtures of these diastereomers. Using the system of equations is quite advantageous, as it is the only mass spectrometric method that has been shown to successfully quantify mixtures of more than two isomers. The utility of the method is demonstrated by successfully quantifying various two and three component mixtures of the diastereomeric monosaccharides. Furthermore, the method is used to quantify the recovery of a single diastereomeric monosaccharide from an acidic resin. Although the multicomponent quantification method described herein is used to quantify mixtures of N-acetylhexosamine diastereomers, it could be applied to any group of isomers, provided distinguishing CID spectra are obtained. This is the first known report of utilizing MS3 product ions for quantification of structural isomeric mixtures.

Acetylgalactosamine↗
PubMed2003

Recognition and quantification of binary and ternary mixtures of isomeric peptides by the kinetic method: metal ion and ligand effects on the dissociation of metal-bound complexes.

The kinetic method is applied to differentiate and quantify mixtures of isomeric tripeptides based on the competitive dissociations of divalent metal ion-bound clusters in an ion trap mass spectrometer. This methodology is extended further to determine compositions of ternary mixtures of the isomers Gly-Gly-Ala (GGA), Ala-Gly-Gly (AGG), and Gly-Ala-Gly (GAG). This procedure also allows to perform chiral quantification of a ternary mixture of optical isomers. The divalent metal ion Ca(II) is particularly appropriate for isomeric distinction and quantification of the isobaric tripeptides Gly-Gly-Leu/Gly-Gly-Ile (GGL/GGI). Among the first-row transition metal ions, Cu(II) yields remarkably effective isomeric differentiation for both the isobaric tripeptides, GGI/GGL using GAG as the reference ligand, and the positional isomers GAG/GGA using GGI as the reference ligand. This is probably due to agostic bonding: alpha-agostic bonding occurs between Cu(II) and GAG and beta-agostic bonding between Cu(II) and GGI, each produces large but different steric effects on the stability of the Cu(II)-bound dimeric clusters. These data form the basis for possible future quantitative analyses of mixtures of larger peptides such as are generated, for example, in combinatorial synthesis of peptides and peptide mimics.

Calcium↗
PubMed2003

Investigation of ion/molecule reactions as a quantification method for phosphorylated positional isomers. an FT-ICR approach.

A rapid and accurate method of quantifying positional isomeric mixtures of phosphorylated hexose and N-acetylhexosamine monosacchrides by using gas-phase ion/molecule reactions coupled with FT-ICR mass spectrometry is described. Trimethyl borate, the reagent gas, reacts readily with the singly charged negative ions of phosphorylated monosaccharides to form two stable product ions corresponding to the loss of one or two neutral molecules of methanol from the original adduct. Product distribution in the ion/molecule reaction spectra differs significantly for isomers phosphorylated in either the 1- or the 6-position. As a result, the percents of total ion current of these product ions for a mixture of the two isomers vary with its composition. In order to determine the percentage of each isomer in an unknown mixture, a multicomponent quantification method is utilized in which the percents of total ion current of the two product ions for each pure monosaccharide phosphate and the mixture are used in a two-equation, two-unknown system. The applicability of this method is demonstrated by successfully quantifying mock mixtures of four different isomeric pairs: Glucose-1-phosphate and glucose-6-phosphate; mannose-1-phosphate and mannose-6-phosphate; galactose-1-phosphate and galactose-6-phosphate; N-acetylglucosamine-1-phosphate and N-acetylglucosamine-6-phosphate. The effects of mixture concentrations and ion/molecule reaction conditions on the quantification are also discussed. Our results demonstrate that this assay is a fast, sensitive, and robust method to quantify isomeric mixtures of phosphorylated monosaccharides.

Fourier Analysis↗
PubMed1999

Noninvasive monitoring of peak filling rate with acoustic quantification echocardiography accurately detects acute cardiac allograft rejection.

BACKGROUND: Acute cardiac allograft rejection is associated with early diastolic dysfunction. The development of chronic rejection is dependent on the frequency and severity of acute rejection episodes. Therefore, early diagnosis and therapy influence long-term survival significantly. For the first time, acoustic quantification, a new echocardiographic technology for on-line measurement of cardiac volumes and their changes, facilitates quantitative assessment of systolic and diastolic function noninvasively. METHODS: Since May 1996, all consecutive patients after cardiac transplantation (n = 94) underwent 475 endomyocardial biopsies and the same number of echocardiographic studies within 6 hours after biopsy before the histological results were available. RESULTS: Nineteen patients showed 23 episodes of acute rejection (ISHLT > or = 2). There was a significant decrease in left ventricular peak filling rate [PFR: end-diastolic volume (EDV)/ second) as a parameter of diastolic function during rejection (2.9 +/- 0.4, n = 23) as compared to PFR measured under nonrejection status (4.5 +/- 0.8; n = 452; p < 0.0001). Most importantly we found that in these 19 patients showing rejection, the PFR was normal in the last examination before rejection, but was significantly reduced during rejection (2.9 +/- 0.4 vs 4.5 +/- 0.7; n = 23, p < 0.0001). After successful rejection therapy, PFR again normalized in all patients, with the exception of 1 patient with steroid-refractory humoral rejection. We calculated sensitivity and specificity for several cutpoints for the event "first rejection" in 15 patients and plotted them in a receiver operating characteristic curve, showing that a PFR > or = 4.0 EDV/second is never associated with treatable rejection. A decrease of PFR of more than 18% from its prevalue of the last biopsy with no rejection increases the accuracy for the diagnosis of rejection significantly. CONCLUSIONS: We conclude that diastolic dysfunction during acute cardiac allograft rejection can be accurately detected by noninvasive measurement of peak filling rate with acoustic quantification echocardiography. Monitoring of this parameter provides reliable discrimination between treatable and nontreatable rejection.

Acute Disease↗
PubMed1997

Improved high-performance liquid chromatographic procedure for the separation and quantification of hydroxytestosterone metabolites.

A reproducible, sensitive high-performance liquid chromatography (HPLC) method has been developed to quantify hydroxytestosterone metabolites. The method features a simple one-step linear gradient of methanol in water (10%-60% methanol) for separation of the testosterone metabolites on a Supelcosil LC-18 column; metabolites are detected at 247 nm. This method provides a distinct advantage over previously developed assays in that the solvent gradient does not contribute to baseline changes throughout the chromatographic run. In this way, the flat baseline markedly improves robustness of the assay and simplifies peak integration and quantification. At the same time, resolution of 15 different steroid metabolites catalyzed by the cytochrome P-450 enzymes are readily separated in rat or mouse liver microsomes. An internal standard, cortexolone, was selected for use based on its structural and spectral similarity to the hydroxytestosterone metabolites, and quantification is based on the molar response for the testosterone/cortexolone peak area ratio. The limit of detection (LOD) is 1 pmol on-column with a limit of quantitation (LOQ) of 4 pmol on-column. The intraday repeatability is approximately 3%. This simplified procedure is straightforward and should greatly facilitate the routine use of the testosterone hydroxylation assay to measure cytochrome P-450 isozyme activity.

Animals↗
PubMed1995

Type II collagen quantification in experimental chondrogenesis.

Type II collagen is an excellent indicator of the cartilage phenotype. Accurate quantification with existing methods requires about 100 micrograms of collagen. We have developed a method which allows for accurate quantification of type II collagen in samples as small as 1 microgram (and sample volumes as little as 1 microliters). Types I and II collagen were pepsin purified, cleaved with cyanogen bromide, and dissolved in sample buffer at concentrations of 0.125-200 micrograms/microliters. Volumes of 1 microliter were analyzed by electrophoresis on microgels. The gels were scanned on a laser densitometer and the ratios of the alpha 1 (II)CB10 to the alpha 1(I)CB7,8 plus alpha 1(II)CB11 determined. The cyanogen bromide-derived (CNBr) peptides could be resolved at concentrations as low as 0.25 microgram/microliter. The ideal working concentration for purified collagens was 1-8 micrograms/microliters. Standard mixtures of both purified and non-purified types I and II collagen were analyzed. At a concentration of 1 microgram/microliter the ratio of the bands referred to above was closely related to the relative proportion of type II collagen, in a polynomial fashion. At 8 micrograms/microliters there was an almost perfect linear relationship. The presence of 15-30% type III collagen had < 5% effect on the measurements of type II collagen. The method is simple, reliable, fast and automated. It should have good potential for application in cartilage research as it permits quantitation of type II collagen in extremely small samples of tissue.

Cartilage, Articular↗
PubMedDate not supplied

Quantification of technetium 99m-labeled sestamibi single-photon emission computed tomography based on mean counts improves accuracy for assessment of relative regional myocardial blood flow: experimental validation in a canine model.

BACKGROUND: Quantification of single-photon emission computed tomographic (SPECT) images is generally based on determination of maximal counts on radial sectors of short-axis slices. We hypothesized that analysis of mean counts may reduce estimation error. METHODS AND RESULTS: We compared quantitative 99mTc-labeled sestamibi (MIBI) SPECT based on maximal myocardial counts with that based on mean myocardial counts for accuracy of quantifying relative regional myocardial perfusion in a canine model of permanent left anterior descending coronary artery occlusion. MIBI and radiolabeled microspheres were injected during left anterior descending coronary artery occlusion. Relative microsphere myocardial blood flow was expressed as a percentage of normal (left circumflex coronary artery territory) blood flow. SPECT imaging was performed in vivo and ex vivo. Relative MIBI uptake on SPECT short-axis slices was quantified with normalized circumferential profiles based on maximal and mean counts. In vivo and ex vivo SPECT relative myocardial count density was compared to relative myocardial blood flow in six dogs. In the comparisons, percent errors in estimating the relative blood flow and relative flow deficit with MIBI SPECT imaging were calculated. There was an excellent correlation between absolute myocardial tissue MIBI activity and regional myocardial blood flow for each of the six dogs (r = 0.90 to 0.98). The correlations between relative myocardial count density on SPECT and relative blood flow for individual sectors were similar for maximal and mean count profiles (maximal, 0.79 to 0.83; mean, 0.77 to 0.82). Comparing the nadirs of in vivo and ex vivo circumferential count profiles, the correlations were slightly better (maximal, 0.82 to 0.91; mean, 0.87 to 0.91). Average percent errors in assessing relative blood flow and relative flow deficit were decreased significantly by use of mean count profiles (p < 0.05). CONCLUSIONS: Relative SPECT count density with either maximal or mean count profiles correlated well with relative myocardial blood flow. Compared with maximal count profiles, quantification with mean count profiles improved estimation of relative flow.

Animals↗