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Accuracy of semiautomated analysis of 3D contrast-enhanced magnetic resonance angiography for detection and quantification of aortoiliac stenoses.

OBJECTIVES: We sought to compare reproducibility and accuracy of semiautomated stenosis detection and quantification in 3D contrast-enhanced magnetic resonance angiography (CE-MRA) images with conventional evaluation of 3D CE-MRA in patients with peripheral arterial disease (PAD) by using intra-arterial digital subtraction angiography (IA-DSA) as standard of reference. METHODS: Twenty-five patients with PAD underwent CE-MRA and IA-DSA. Three blinded observers independently evaluated CE-MRA datasets for stenoses in the aortoiliac region using semiautomated computer analysis. Semiautomated measurements were compared with conventional measurements of stenosis on CE-MRA datasets, as measured by 3 other independent observers blinded to all other measurements. Interobserver agreement was quantified using kappa (kappa) and intraclass correlation coefficients (ICCs). Sensitivity and specificity were determined for both semiautomated and conventional measurements. RESULTS: Semiautomated measurements were successful in 124 of the 125 arterial segments. The sensitivity of semiautomated measurements was 89% for all observers; specificity varied between 87% and 89%. For conventional measurements of CE-MRA, sensitivity varied between 79% and 86%; specificity was between 86% and 96%. There was good interobserver agreement between all readers for semiautomated measurements (combined kappa for all 3 observers together = 0.78; ICC = 0.82), as well as for conventional measurements (combined kappa = 0.70; and ICC = 0.83). Differences between ICCs, combined kappa values, and accuracy of both measurements were not significant (all P > 0.05). CONCLUSION: Semiautomated analysis of aortoiliac 3D CE-MRA has the same high accuracy for detection and quantification of stenoses as conventional readings of CE-MRA.

Aged↗

Quantification of aortic valve calcification using multislice spiral computed tomography: comparison with atomic absorption spectroscopy.

OBJECTIVES: Multislice spiral computed tomography (MSCT) allows the in vivo detection of valvular calcification. The aim of this study was to validate the quantification of aortic valve calcification (AVC) by MSCT with in vitro measurements by atomic absorption spectroscopy. METHODS: In 18 patients with severe aortic stenosis, 16 detector row MSCT (SOMATOM Sensation 16, Siemens, Forchheim, Germany with scan parameters as follows: 420 milliseconds tube rotation time, 12 x 0.75 mm collimation, tube voltage 120 KV) was performed before aortic valve replacement. Images were reconstructed at 60% of the RR interval with an effective slice thickness of 3 mm and a reconstruction increment of 2 mm. AVC was assessed using Agatston AVC score, mass AVC score, and volumetric AVC score. After valve replacement, the calcium content of the excised human stenotic aortic valves was determined in vitro using atomic absorption spectroscopy. RESULTS: The mean Agatston AVC score was 3,842 +/- 1,790, the mean volumetric AVC score was 3,061 +/- 1,406, and mass AVC score was 888 +/- 492 as quantified by MSCT. Atomic absorption spectroscopy showed a mean true calcification mass (Ca5(PO4)3OH) of 19 +/- 8 mass%. There was a significant correlation between in vivo AVC scores determined by MSCT and in vitro mean true calcification mass (r = 0.74, P = 0.0004 for mass AVC score, r = 0.79, P = 0.0001 for volumetric AVC score and r = 0.80, P = 0.0001 for Agatston AVC score) determined by atomic absorption spectroscopy. Linear regression analysis showed a significant association between the degree of hydroxyapatite (given in mass%) in the aortic valve and the degree of AVC (R = 0.74, F = 19.6, P = 0.0004 for mass AVC score, R = 0.80, F = 29.3, P = 0.0001 for Agatston AVC score and R = 0.79, F = 27.3, P = 0.0001 for volumetric AVC score) assessed by MSCT. CONCLUSION: MSCT allows accurate in vivo quantification of aortic valve calcifications.

Aged↗

Digital quantification of mutant DNA in cancer patients.

PURPOSE OF REVIEW: The accumulation of somatic mutations is the major driving force for tumorigenesis. These mutations uniquely differentiate tumor cells from their normal counterparts. Mutations within tumor cells and mutant DNA released by tumor cells into blood, lymph, stool, tissues and other bodily compartments can thereby be used for cancer detection. Here we discuss technologies available for the detection and quantification of mutant DNA in clinical samples and the value of such measurements for patient management. RECENT FINDINGS: Conventional mutation detection technologies are either qualitative or only roughly estimate the abundance of mutant DNA molecules. Recently-developed approaches, however, use single molecule counting to determine the genotype of each individual member of a DNA population, providing a more accurate and precise digital output. SUMMARY: In this review, we discuss the clinical utility of mutant DNA quantification in cancer patients in the context of recent technical advances made in digital mutation detection.

DNA Mutational Analysis↗

Quantification of the antiviral effect of interferon by immunoassay of vesicular stomatitis virus proteins.

A simple solid-phase immunoassay for quantification of vesicular stomatitis virus (VSV) is described. Infected cultures are lysed with deoxycholate. Samples of the lysates are transferred to PVC immunoassay plates and the amount of virus protein adsorbed to the plates is then quantified by sequential incubation with antiserum against VSV proteins and 125I-labelled Protein A. The decrease of VSV protein in interferon (IFN)-treated cultures is correlated with inhibition of formation of infectious virions; its quantification therefore allows accurate measurement of the antiviral effect. The applicability of the immunoassay for measuring the virus yield is not restricted to cells exhibiting a virus cytopathic effect. Moreover, since the decrease of virus protein is obtained at IFN concentrations lower than those that reduce cell killing by the virus, the assay provides a more sensitive measure for the IFN effect than that obtained by 'cytopathic effect inhibition' assays.

Cell Line↗

Quantification of human cytomegalovirus DNA using the polymerase chain reaction.

The important goal of developing quantitative assays for viral nucleic acids in clinical samples has been achieved for human cytomegalovirus (HCMV) by using a modified polymerase chain reaction (PCR). A control PCR target sequence was constructed by PCR mutagenesis to allow the post-amplification quantification of HCMV DNA. The control region was identical to a naturally occurring sequence within the glycoprotein B (gB) coding part of the virus genome, except that a unique restriction site, introduced by the aforementioned mutagenesis step, allowed post-amplification differentiation of control/non-control target amplified product. This technique was initially validated using known amounts of cloned control/non-control target DNA, and was found to be sufficiently sensitive to allow the quantification of a range of 10 to 10(6) genome equivalents of virus. The method was applied to urine samples of congenitally infected infants for which infectious virus titres were available. The results obtained demonstrated that the number of infectious virions determined by conventional cell culture represented a small proportion of the HCMV genome present in the samples, as assessed by the quantitative PCR methodology.

Base Sequence↗

Monochrome LightCycler PCR assay for detection and quantification of five common species of Candida and Aspergillus.

Invasive fungal pathogens, especially in immunocompromised hosts, can result in life-threatening infections. Current laboratory/radiological methods for fungal identification are time-consuming and lack sensitivity and specificity. A monochrome, multiplex, real-time PCR assay for the identification and quantification of Candida albicans, Candida krusei, Candida tropicalis, Aspergillus flavus and Aspergillus fumigatus is described here. Detection of each of these fungi was specific and demonstrated 100 % concordance with biochemical/culture identification in all 60 isolates tested. Samples from 16 febrile neutropenic patients with haematological malignancies were also analysed and the utility of the assay in clinical samples was reconfirmed without false-negative results. The sensitivity of this assay was 0.1 pg fungal genomic DNA, corresponding to three cells, for C. albicans, C. krusei, C. tropicalis and A. flavus, and 0.01 pg fungal genomic DNA, i.e. less than one cell, for A. fumigatus. The analysis allows a low-cost, simple, rapid and sensitive alternative for clinical identification and quantification of these five common fungal species.

Aspergillosis↗

Detection and quantification of Legionella pneumophila DNA in serum: case reports and review of the literature.

Legionella pneumonia can be difficult to diagnose. Existing laboratory tests all have shortcomings, especially the ability to diagnose all Legionella spp. at an early stage. Detection of Legionella DNA in serum can be a valuable tool for the diagnosis of Legionnaires' disease (LD). This report describes two patients with LD diagnosed by PCR using serum samples. In addition, quantification of L. pneumophila DNA using real-time PCR during the course of illness was carried out. The results obtained mirrored both the clinical condition and C-reactive protein values during the course of the illness. Quantification of Legionella DNA in serum using real-time PCR could be a valuable tool to monitor the effects of antimicrobial therapy in patients with LD.

Adult↗

Quantification, sources, and control of ammonia emissions in the Czech Republic.

The exact quantification of ammonia (NH3) emissions is the basic presumption for the fulfillment of obligations set by the CLRTAP (Convention on Long Range Transboundary Air Pollution) Protocol which was signed by the Czech Republic in 1999. Most NH 3 emissions in the Czech Republic are produced during the breeding of cattle, pigs, and poultry; therefore, determinating emission factors for these kinds of animals by studying their total number, type of breeding, and subsequent disposal of manure is the solution to the problem of NH 3 emissions quantification. This paper summarizes the results of 4 years of research in this area, determining the emission factors and ways of decreasing emissions from the breeding of cattle, pigs, and poultry.

Agriculture↗

Fully automatic quantification of microarray image data.

DNA microarrays are now widely used to measure expression levels and DNA copy number in biological samples. Ratios of relative abundance of nucleic acids are derived from images of regular arrays of spots containing target genetic material to which fluorescently labeled samples are hybridized. Whereas there are a number of methods in use for the quantification of images, many of the software systems in wide use either encourage or require extensive human interaction at the level of individual spots on arrays. We present a fully automatic system for microarray image quantification. The system automatically locates both subarray grids and individual spots, requiring no user identification of any image coordinates. Ratios are computed based on explicit segmentation of each spot. On a typical image of 6000 spots, the entire process takes less than 20 sec. We present a quantitative assessment of performance on multiple replicates of genome-wide array-based comparative genomic hybridization experiments. By explicitly identifying the pixels in each spot, the system yields more accurate estimates of ratios than systems assuming spot circularity. The software, called, runs on Windows platforms and is available free of charge for academic use.

Breast Neoplasms↗

Quantification of multiple gene expression in individual cells.

Quantitative gene expression analysis aims to define the gene expression patterns determining cell behavior. So far, these assessments can only be performed at the population level. Therefore, they determine the average gene expression within a population, overlooking possible cell-to-cell heterogeneity that could lead to different cell behaviors/cell fates. Understanding individual cell behavior requires multiple gene expression analyses of single cells, and may be fundamental for the understanding of all types of biological events and/or differentiation processes. We here describe a new reverse transcription-polymerase chain reaction (RT-PCR) approach allowing the simultaneous quantification of the expression of 20 genes in the same single cell. This method has broad application, in different species and any type of gene combination. RT efficiency is evaluated. Uniform and maximized amplification conditions for all genes are provided. Abundance relationships are maintained, allowing the precise quantification of the absolute number of mRNA molecules per cell, ranging from 2 to 1.28 x 10(9) for each individual gene. We evaluated the impact of this approach on functional genetic read-outs by studying an apparently homogeneous population (monoclonal T cells recovered 4 d after antigen stimulation), using either this method or conventional real-time RT-PCR. Single-cell studies revealed considerable cell-to-cell variation: All T cells did not express all individual genes. Gene coexpression patterns were very heterogeneous. mRNA copy numbers varied between different transcripts and in different cells. As a consequence, this single-cell assay introduces new and fundamental information regarding functional genomic read-outs. By comparison, we also show that conventional quantitative assays determining population averages supply insufficient information, and may even be highly misleading.

Base Sequence↗

Visualization, quantification and therapeutic evaluation of angiogenic vessels in cancer by synchrotron microangiography.

The usefulness of a synchrotron microangiography system for depicting, quantitating and therapeutically evaluating angiogenic vessels in cancer is illustrated. In 20 mice transplanted with murine colon cancer, sequential changes in the angiogenic vessels were determined by using synchrotron microangiography, using changes in tumor volume for reference. This system allowed the depiction and quantification of angiogenic vessels in the period from one to four weeks after transplantation. The effects of antiangiogenic therapy were evaluated by using a neutralizing antibody against vascular endothelial growth factor. The neutralizing antibody partially suppressed angiogenesis and tumor growth. Synchrotron microangiography is shown to be useful for the depiction, quantification and evaluation of angiogenic vessels in cancer.

Journal Article↗

A viewpoint determination system for stenosis diagnosis and quantification in coronary angiographic image acquisition.

This paper describes the usefulness of computer assistance in the acquisition of "good" images for stenosis diagnosis and quantification in coronary angiography. The system recommends the optimal viewpoints from which stenotic lesions can be observed clearly based on images obtained from initial viewpoints. First, the viewpoint dependency of the apparent severity of a stenotic lesion is experimentally analyzed using software phantoms in order to show the seriousness of the problem. The implementation of the viewpoint determination system is then described. The system provides good user-interactive tools for the semiautomated estimation of the orientation and diameter of stenotic segments and the three-dimensional (3-D) reconstruction of vessel structures. Using these tools, viewpoints that will not give rise to foreshortening and vessel overlap can be efficiently determined. Experiments using real coronary angiograms show the system to be capable of the reliable diagnosis and quantification of stenosis.

Coronary Angiography↗

Feature extraction and quantification of the variability of dynamic performance profiles due to the different sagittal lift characteristics.

Investigation of manual material handling (MMH) tasks, such as lifting, requires the quantification of the various kinematic and kinetic parameters of performance for assessment of the functional capacity and/or task demand profiles. Traditional statistical descriptive analyses usually involve computing the summary statistics (maximum, minimum, mean, and/or range) of the resulting performance parameters over the cycle duration (i.e., lifting/lowering cycle). Consequently, the significant information content of the time-varying signals is diminished, limiting the sensitivity of subsequent hypothesis testing procedures. The present study developed a methodology for representing and quantifying performance data variability of the kinematic and kinetic motion profiles due to the different lift characteristics (load, mode, and speed) during MMH tasks while capturing the temporal characteristics. Using a database of motion profiles from a manual lifting experiment, the Karhunen-Loeve Expansion (KLE) feature extraction technique was shown to be quite effective for representing the various motion profiles. The number of basis vectors (eigenvectors) and corresponding coefficients needed for accurate representation were substantially smaller than the original data set, resulting in data compression. Moreover, the effects of lift characteristics were investigated using analysis of variance techniques that recognize the vectorial constitution of the waveforms. The application of these techniques will enable the quantification of highly phasic profiles and enhance the ability to document the effect of intervening measures such as educational or physical training/exercise on the kinematic and kinetic patterns of performance. Additionally, the differential influence of lift characteristics on the variability of performance during different phases of lifting and lowering provides added resolution in the analysis of MMH tasks.

Adult↗

Movement quantification in epileptic seizures: a new approach to video-EEG analysis.

It is common that epileptic seizures induce uncoordinated movement in a patient's body. This movement is a relevant clinical factor in seizure identification. Nevertheless, quantification of this information has not been an object of much attention from the scientific community. In this paper, we present our effort in developing a new approach to the quantification of movement patterns in patients during epileptic seizures. We attach markers at landmark points of a patient's body and use a camera and a commercial video-electroencephalogram (EEG) system to synchronously register EEG and video during seizures. Then, we apply image-processing techniques to analyze the video frames and extract the trajectories of those points that represent the course of the quantified movement of different body parts. This information may help clinicians in seizure classification. We describe the framework of our system and a method of analyzing video in order to achieve the proposed goal. Our experimental results show that our method can reflect quantified motion patterns of epileptic seizures, which cannot be accessed by means of traditional visual inspection of video recordings. We were able, for the first time, to quantify the movement of different parts of a convulsive human body in the course of an epileptic seizure. This result represents an enhanced value to clinicians in studying seizures for reaching a diagnosis.

Algorithms↗

In vivo quantification of retraction deformation modeling for updated image-guidance during neurosurgery.

The use of coregistered preoperative anatomical scans to provide navigational information in the operating room has greatly benefited the field of neurosurgery. Nonetheless, it has been widely acknowledged that significant errors between the operating field and the preoperative images are generated as surgery progresses. Quantification of tissue shift can be accomplished with volumetric intraoperative imaging; however, more functional, lower cost alternative solutions to this challenge are desirable. We are developing the strategy of exploiting a computational model driven by sparse data obtained from intraoperative ultrasound and cortical surface tracking to warp preoperative images to reflect the current state of the operating field. This paper presents an initial quantification of the predictive capability of the current model to computationally capture tissue deformation during retraction in the porcine brain. Performance validation is achieved through comparisons of displacement and pressure predictions to experimental measurements obtained from computed tomographic images and pressure sensor recordings. Group results are based upon a generalized set of boundary conditions for four subjects that, on average, account for at least 75% of tissue motion generated during interhemispheric retraction. Individualized boundary conditions can improve the degree of data-model match by 10% or more but warrant further study. Overall, the level of quantitative agreement achieved in these experiments is encouraging for updating preoperative images to reflect tissue deformation resulting from retraction, especially since model improvements are likely as a result of the intraoperative constraints that can be applied through sparse data collection.

Animals↗

Using recurrence quantification analysis determinism for noise removal in cardiac optical mapping.

Selecting signal processing parameters in optical imaging by utilizing the change in Determinism, a measure introduced in Recurrence Quantification Analysis, provides a novel method using the change in residual noise Determinism for improving noise quantification and removal across signals exhibiting disparate underlying tissue pathologies. The method illustrates an improved process for selecting filtering parameters and how using measured signal-to-noise ratio alone can lead to improper parameter selection.

Algorithms↗

Contrast echocardiography for pulmonary blood volume quantification.

Pulmonary blood volume quantification is important both for diagnosis and for monitoring of the circulatory system. It requires employment of transpulmonary indicator dilution techniques, which are very invasive due to the need for double catheterization. This paper presents a new minimally invasive technique for blood volume quantification. An ultrasound contrast agent bolus is injected peripherally and detected by an ultrasound transducer in the central circulation. Several echocardiographic views permit simultaneous detection of contrast in different cardiac cavities and central vessels, and acoustic backscatter measurements produce multiple indicator dilution curves (IDCs). Contrast mean-transit-time differences are derived from the IDC analysis and multiplied times cardiac output for the assessment of blood volumes between different detection sites. For pulmonary blood volume estimates, the right ventricle and the left atrium IDCs are measured. The mean transit time of the IDC is estimated by specific modelling. The Local Density Random Walk and the First Passage Time models were tested for IDC interpolation and interpretation. The system was validated in vitro for a wide range of flows. The results show very accurate volume measurements. The volume estimate determination coefficient is greater than 0.999 for both model fits. A preliminary study in patients shows promising results.

Algorithms↗

A new formalism for the quantification of tissue perfusion by the destruction-replenishment method in contrast ultrasound imaging.

A new formalism is presented for the destruction-replenishment perfusion quantification approach at low mechanical index. On the basis of physical considerations, best-fit methods should be applied using perfusion functions with S-shape characteristics. These functions are first described for the case of a geometry with a single flow velocity, then extended to the case of vascular beds with blood vessels having multiple flow velocity values and directions. The principles guiding the analysis are, on one hand, a linearization of video echo signals to overcome the log-compression of the imaging instrument, and, on the other hand, the spatial distribution of the transmit-receive ultrasound beam in the elevation direction. An in vitro model also is described; it was used to confirm experimentally the validity of the approach using a commercial contrast agent. The approach was implemented in the form of a computer program, taking as input a sequence of contrast-specific images, as well as parameters related to the ultrasound imaging equipment used. The generated output is either flow-parameter values computed in regions-of-interest, or parametric flow-images (e.g., mean velocity, mean transit time, mean flow, flow variance, or skewness). This approach thus establishes a base for extracting information about the morphology of vascular beds in vivo, and could allow absolute quantification provided that appropriate instrument calibration is implemented.

Algorithms↗