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Biomedical subjects

F Frouin

Publications and source records attributed to F Frouin.

At least 19 recordsLinked to original sources

A posteriori respiratory gating in contrast ultrasound for assessment of hepatic perfusion.

An original strategy is proposed to minimize the impact of respiratory motion on hepatic contrast-enhanced ultrasound studies. It is based on the a posteriori triggering of dynamic image sequences. It was tested on perfusion studies acquired with a high temporal resolution (8 images s-1) to enable parametric imaging. A respiratory component was first estimated by independent component analysis. The estimation of the local minima and maxima of this curve enabled us to select two subsets of frames, corresponding to the end-of-inspiration plane and to the end-of-expiration plane. Both subsets were simultaneously analysed using factor analysis of medical image sequences. This method identified the main contrast uptake kinetics and their associated localizations. The global strategy was validated firstly on a simulated study and then applied to 11 patients' studies. In both cases, the frame selection was judged relevant and a necessary preliminary step before applying methods of parametric imaging. In conclusion, the a posteriori gating method that is proposed is a first step towards local quantification of hepatic contrast-enhanced ultrasound studies.

Computer Simulation↗

Assessment of left ventricular contraction by parametric analysis of main motion (PAMM): theory and application for echocardiography.

The computerized study of the regional contraction of the left ventricle has undergone numerous developments, particularly in relation to echocardiography. A new method, parametric analysis of main motion (PAMM), is proposed in order to synthesize the information contained in a cine loop of images in parametric images. PAMM determines, for the intensity variation time curves (IVTC) observed in each pixel, two amplitude coefficients characterizing the continuous component and the alternating component; the variable component is generated from a mother curve by introducing a time shift coefficient and a scale coefficient. Two approaches, a PAMM data driven and a PAMM model driven (simpler and faster), are proposed. On the basis of the four coefficients, an amplitude image and an image of mean contraction time are synthesized and interpreted by a cardiologist. In all cases, both PAMM methods allow better IVTC adjustment than the other methods of parametric imaging used in echocardiography. A preliminary database comprising 70 segments is scored and compared with the visual analysis, taken from a consensus of two expert interpreters. The levels of absolute and relative concordance are 79% and 97%. PAMM model driven is a promising method for the rapid detection of abnormalities in left ventricle contraction.

Algorithms↗

Analysis of the distribution of MRI contrast agents in the livers of small animals by means of complementary microscopies.

BACKGROUND: Magnetic resonance imaging (MRI) contrast agents contain magnetic molecules such as iron (Fe) or gadolinium (Gd) that are injected in vivo into rats or mice to study their distribution inside the liver. Fluorescent europium (Eu) can be used as a model of Gd to obtain comparable information of this distribution of corresponding contrast agents. In a similar approach, Fe can be attached to Texas Red and used as a model of ferumoxides and be detected by fluorescence. METHODS: To combine and compare the advantages of different microscopic imaging modes, characterization studies were carried out by means of a confocal laser scanning microscope (CLSM), a secondary ion mass spectrometric (SIMS) microscope, and an electron energy loss spectrometric (EELS) microscope. In the case of CLSM, the locations of fluorescent signals inside preparations were determined by factor analysis of biomedical image sequences (FAMIS) and selection of image sequences at emission. RESULTS: By CLSM and FAMIS, we distinguished chelated Eu and Texas Red attached to Fe. By SIMS microscopy, we distinguished Eu and Gd of chlorides and chelates and Fe of a ferumoxide. By EELS microscopy, we distinguished Eu and Gd of chlorides. CONCLUSIONS: Analysis of compounds inside correlative specimens by means of CLSM, SIMS, and EELS microscopes provided complementary results.

Animals↗

Local reconstruction of stenosed sections of artery using multiple MRA acquisitions.

A method for reconstructing magnetic resonance angiography (MRA) volumes from successive acquisitions is described. The method is based on double oblique acquisitions of highly anisotropic MRA volumes, each of which corresponds to reduced k-space filling. These partial k-spaces are then combined to obtain a 3D k-space adapted to the frequency spread of the angiographic image of the stenosis. The SNR-resolution compromise of MRA is thus improved by focusing the acquisition on the most relevant k-space regions. The reconstruction is performed directly in k-space by averaging the partial k-spaces. The feasibility of the method was demonstrated in studies on a Lucite stenosis phantom, on MRAs of carotid arteries using three bolus injections, and on MRAs of renal arteries using a single contrast injection.

Carotid Arteries↗

Reduced capillary perfusion and permeability in human tumour xenografts treated with the VEGF signalling inhibitor ZD4190: an in vivo assessment using dynamic MR imaging and macromolecular contrast media.

We describe the use of perfusion-permeability magnetic resonance imaging (ppMRI) to study hemodynamic parameters in human prostate tumor xenografts, following treatment with the vascular endothelial growth factor-A (VEGF) receptor tyrosine kinase inhibitor, ZD4190. Using a macromolecular contrast agent (P792), a fast MR imaging protocol and a compartmental data analysis, we were able to demonstrate a significant simultaneous reduction in tumor vascular permeability, tumor vascular volume and tumor blood flow (43%, 30% and 42%, respectively) following ZD4190 treatment (100 mg/kg orally, 24 h and 2 h prior to imaging). This study indicates that MR imaging can be used to measure multiple hemodynamic parameters in tumors, and that tumor vascular permeability, volume and flow, can change in response to acute treatment with a VEGF signaling inhibitor.

Angiogenesis Inhibitors↗

[Image analysis in contrast echocardiography].

The visualization of contrast agents in echocardiography is obtained with complex acquisition sequences and their analysis is dependent on operator experience. In order to be more accessible, images analysis is essential and the main objective is to integrate qualitative and quantitative data in one single image.

Contrast Media↗

[Value of factor analysis in a wall motion study: preliminary application in the detection of left ventricular segmental contraction ischemic disorders by echocardiography].

PURPOSE: Factor Analysis of Medical Image Sequences (FAMIS) was tested to study the regional wall motion of the left ventricle at echocardiography. MATERIALS AND METHODS: FAMIS analyzed the time signal curves of each pixel. One flat curve and one curve describing the contraction-relaxation of the left ventricle were first estimated. The contributions of the curve of each pixel to the two previous curves were computed, yielding two "factor images". The spatial distribution of positive and negative coefficients of the second factor image was analyzed. The evaluation was carried out on 222 segments (20 patients, 18 parasternal short-axis views, 17 apical four-chamber views, and 15 apical two-chamber views). A first echocardiographer reviewed the factor images and the reading was compared to the conventional reading of the cine-loops by two other echocardiographers. Each segment was scored as normal, hypokinetic, akinetic, or dyskinetic. RESULTS: On normal segments, the positive coefficients of the second factor image were on the inner side, the negative coefficients were on the outer side. Dyskinesis yields the opposite pattern. Hypokinesis and akinesis give intermediate images. An absolute concordance was obtained on 71.2% of all segments between the two types of reading. Larger discrepancies were found for akinetic and hypokinetic segments. CONCLUSION: FAMIS is a promising tool to study regional wall motion of the left ventricle.

Echocardiography, Doppler, Color↗

Adaptive and self-evaluating registration method for myocardial perfusion assessment.

With the advent of ultra-fast MRI, it is now possible to assess non-invasively regional myocardial perfusion with multislice coverage and sub-second temporal resolution. First-pass contrast enhanced studies are acquired with ECG-triggering and breath holding. Nevertheless, some respiratory induced movements still remain. Myocardial perfusion can be assessed locally by parametric imaging methods such as Factor Analysis of Medical Image Sequences (FAMIS), provided that residual motion can be corrected. An a posteriori registration method implemented in the image domain is proposed. It is based on an adaptive registration model of the heart combining three elementary shapes (left ventricle, right ventricle and pericardium). The registration procedure is performed on a potential map derived from the distance map. To evaluate the quality of the registration procedure a superimposition score between the registration model and the contour automatically extracted in the sequence is proposed. Rigid transformation hypotheses and registration analysis provide an efficient and automatic method which allows the rejection of outlier images, such as: out of synchronisation images, out of plane acquisitions. When compared to a manual registration method, this approach reduces processing time and requires a minimal intervention from the operator. The proposed method performs registration with a subpixel accuracy. It has been successfully applied to simulated images and clinical data. It should facilitate the use of MR first-pass perfusion studies in clinical practice.

Humans↗

Early changes in liver perfusion caused by occult metastases in rats: detection with quantitative CT.

PURPOSE: To determine whether computed tomography (CT) can depict liver hemodynamic changes caused by occult hepatic micrometastases in rat. MATERIALS AND METHODS: Liver micrometastases (mean diameter, 500 micrometer +/- 300) were produced in seven BD IX rats by injecting 10(7) DHDK12 PROb colorectal carcinoma cells into the spleen. Macrometastases (mean diameter, 7 mm +/- 3) were produced in four other rats. Five normal rats were studied as controls. CT images were obtained every 300 msec for 30 seconds during the injection of 1 mL per kilogram of body weight of contrast medium. The time-attenuation curves of the aorta, portal vein, and liver were used to calculate liver perfusion with a deconvolution model designed for the dual blood supply. RESULTS: Micrometastases in an apparently normal liver caused a 34% decrease in portal blood flow and a 25% increase in the mean transit time for the blood to pass through the liver. These findings suggest increased resistance in the sinusoidal capillaries. Similar but greater changes were found in the macrometastases. CONCLUSION: Occult liver micrometastases in rats generate changes in liver perfusion that can be detected with CT.

Animals↗

Confocal analysis of phosphatidylserine externalization with the use of biotinylated annexin V revealed with streptavidin-FITC, -europium, -phycoerythrin or -Texas Red in oxysterol-treated apoptotic cells.

OBJECTIVE: To analyze externalization of phosphatidylserine via annexin V on apoptotic cells by laser scanning confocal microscopy and factor analysis of biomedical image sequences (FAMIS). STUDY DESIGN: Streptavidin-fluorescein isothiocyanate (FITC), -europium (Eu), -phycoerythrin (PE) and -Texas Red (TR) were chosen to reveal the binding of biotinylated annexin V on apoptotic U937 human leukemic cells and ECV-304 human endothelial cells induced under treatment with 7-ketocholesterol or 7 beta-hydroxycholesterol. Excitation of each fluorochrome was obtained by selection of specific lines (351 + 364 nm, 488 nm) of the argon laser of a confocal microscope. Temporal and spectral series were performed to characterize each fluorochrome. FAMIS was applied to these series to estimate images corresponding to stains. RESULTS: Each fluorochrome was clearly distinguished, and images showed localization of phosphatidylserine, which was improved by image analysis. CONCLUSION: On apoptotic cells it is possible to analyze differences in the improved visualization of phosphatidylserine in series processed by FAMIS with the use of biotinylated annexin V revealed with streptavidin-FITC, -Eu, -PE or -TR.

Annexin A5↗

Confocal image characterization of human papillomavirus DNA sequences revealed with Eu in HeLa cell nuclei stained with Hoechst 33342.

OBJECTIVE: To visualize and localize specific viral DNA sequences revealed with Eu by fluorescence in situ hybridization, confocal laser scanning microscopy (CLSM) and factor analysis of biomedical image sequences (FAMIS). STUDY DESIGN: Human papillomavirus DNA (HPV-DNA) was identified in HeLa cells with biotinylated DNA probes recognizing HPV-DNA types 16/18. DNA-DNA hybrids were revealed by a three-step immunohistochemical amplification procedure involving an antibiotin mouse monoclonal antibody, a biotinylated goat antimouse polyclonal antibody and streptavidin-Eu. Cell nuclei were counterstained with Hoechst 33342. Image sequences were obtained using a CLSM that made possible ultraviolet excitation. The location of fluorescent signals inside cellular preparations was determined by FAMIS and selection of filters at emission. Image sequences were summarized into a reduced number of images, or factor images, and curves, or factors. Factors estimate spectral or temporal patterns and depth emission profiles. Factor images correspond to spatial distributions of the different factors. RESULTS: We distinguished between Eu corresponding to HPV-DNA hybridization signals and nuclear staining by taking into account differences in their spectral and temporal patterns and (using their decay rates). CONCLUSION: FAMIS, together with CLSM and Eu, made possible the detection and characterization of viral papillomavirus DNA sequences in HeLa cells.

Base Sequence↗

Validation of myocardial perfusion reserve measurements using regularized factor images of H(2)(15)O dynamic PET scans.

UNLABELLED: The use of H(2)(15)O PET scans for the measurement of myocardial perfusion reserve (MPR) has been validated in both animal models and humans. Nevertheless, this protocol requires cumbersome acquisitions such as C(15)O inhalation or (18)F-FDG injection to obtain images suitable for determining myocardial regions of interest. Regularized factor analysis is an alternative method proposed to define myocardial contours directly from H(2)(15)O studies without any C(15)O or FDG scan. The study validates this method by comparing the MPR obtained by the regularized factor analysis with the coronary flow reserve (CFR) obtained by intracoronary Doppler as well as with the MPR obtained by an FDG acquisition. METHODS: Ten healthy volunteers and 10 patients with ischemic cardiopathy or idiopathic dilated cardiomyopathy were investigated. The CFR of patients was measured sonographically using a Doppler catheter tip placed into the proximal left anterior descending artery. The mean velocity was recorded at baseline and after dipyridamole administration. All subjects underwent PET imaging, including 2 H(2)(15)O myocardial perfusion studies at baseline and after dipyridamole infusion, followed by an FDG acquisition. Dynamic H(2)(15)O scans were processed by regularized factor analysis. Left ventricular cavity and anteroseptal myocardial regions of interest were drawn independently on regularized factor images and on FDG images. Myocardial blood flow (MBF) and MPR were estimated by fitting the H(2)(15)O time-activity curves with a compartmental model. RESULTS: In patients, no significant difference was observed among the 3 methods of measurement-Doppler CFR, 1.73 +/- 0.57; regularized factor analysis MPR, 1.71 +/- 0.68; FDG MPR, 1.83 +/- 0.49-using a Friedman 2-way ANOVA by ranks. MPR measured with the regularized factor images correlated significantly with CFR (y = 1.17x - 0.30; r = 0.97). In the global population, the regularized factor analysis MPR and FDG MPR correlated strongly (y = 0.99x; r = 0.93). Interoperator repeatability on regularized factor images was 0.126 mL/min/g for rest MBF, 0.38 mL/min/g for stress MBF, and 0.34 for MPR (19% of mean MPR). CONCLUSION: Regularized factor analysis provides well-defined myocardial images from H(2)(15)O dynamic scans, permitting an accurate and simple measurement of MPR. The method reduces exposure to radiation and examination time and lowers the cost of MPR protocols using a PET scanner.

Analysis of Variance↗

Confocal multilaser focusing and single-laser characterization of ultraviolet excitable stains of cellular preparations.

BACKGROUND: The aims of this study were (1) to realign cellular preparations when spots and structures are excited by different lasers of a confocal laser scanning microscope (multilaser studies); (2) to avoid the use of realigment methods by selecting fluorochromes that can be excited by only one laser (single-laser experiments). METHODS: In multilaser studies, we used propidium iodide fluorescent beads, as well as tetramethyl rhodamine isothiocyanate (TRITC), fluorescein isothiocyanate (FITC), and 4'-6 diamidino-2-phenylindole (DAPI)-stained human cancer lines. They were excited using HeNe, argon, and ultraviolet (UV) argon laser lines of a confocal laser scanning microscope. Single-laser experiments using UV excitation only were performed using europium as a model for magnetic resonance paramagnetic contrast agents. Nuclei of human cancer lines and tissue were counterstained by DAPI and cytoplasms were labeled with ELF-97 substrates. Factor analysis of medical images (FAMIS) and correlation methods were used to realign shifted images, focus images, and characterize each fluorochrome when necessary. RESULTS: In multilaser studies, superimposition of factor images corrected Z shifts and correlation methods provided X, Y correction values. In single-laser experiments, each fluorochrome was clearly distinguished in the group of fluorochromes. Estimated images in both studies showed colocalizations of structures. CONCLUSIONS: It is possible to characterize differences in the focus and alignment of fluorescent probes and to correct them. It is also possible to study colocalization of UV excitable fluorochromes (DAPI, ELF-97, europium) in cellular and tissular preparations via multilaser or single-laser experiments.

Europium↗

Improved estimation of velocity and flow rate using regularized three-point phase-contrast velocimetry.

We improved the three-point phase-contrast method by regularization of MR velocity data after acquisition of a low velocity-to-noise ratio (VNR) velocity image and a high VNR aliased velocity image. The phase unwrapping algorithm is based on the assumed correlation of the velocity of adjacent flow voxels on the low VNR and the unaliased high VNR images. We used Fourier encoding with eight velocity-encoding gradient steps to obtain reference velocity images of the aorta from five subjects (274 images) and compared them with the phase-contrast and three-point phase-contrast images with and without regularization. The VNR of the regularized velocity image was improved by 9.1 dB and the VNR of the three-point phase-contrast velocity image was improved by 0.7 dB with respect to the low first moment velocity image. Corresponding improvements of 9 dB and 3.7 dB were obtained for the estimations of instantaneous flow rate. Magn Reson Med 44:122-128, 2000.

Adult↗

Factor analysis of confocal image sequences of human papillomavirus DNA revealed with fast red in cervical tissue sections stained with TOTO-iodide.

OBJECTIVE: To visualize and localize specific DNA sequences by fluorescence in situ hybridization, confocal laser scanning microscopy (CLSM) and factor analysis of biomedical image sequences (FAMIS). STUDY DESIGN: Human papillomavirus (HPV) DNA was identified in cervical tissue sections with biotinylated DNA probes recognizing the whole genome of HPV DNA types 18 and 16, and DNA-DNA hybrids were revealed by streptavidin-alkaline phosphatase and Fast Red (FR). Cell nuclei were counterstained with TOTO-iodide. Image sequences were obtained using successive dynamic or spectral sequences of images on different optical slices from CLSM. The location of fluorescent signals inside tissue preparations was determined by FAMIS and/or selection of filters at emission. Image sequences were summarized into a reduced number of images, called "factor images," and curves, called "factors." Factors estimate spectral patterns and depth emission profiles. Factor images correspond to spatial distributions of the different factors. RESULTS: We distinguished between FR and nucleus staining in HPV DNA hybridization signals by taking into account differences in their spectral patterns and improved visualization by taking into account differences in their focus (depth emission profiles). CONCLUSION: FAMIS, together with CLSM, made possible the detection and characterization of HPV DNA sequences in cells of cervical tissue sections.

Azo Compounds↗

Four-dimensional factor analysis of confocal image sequences (4D-FAMIS) to detect and characterize low copy numbers of human papillomavirus DNA by FISH in HeLa and SiHa cells.

Visualization and localization of specific DNA sequences were performed by fluorescence in situ hybridization, confocal laser scanning microscopy (CLSM), and four-dimensional factor analysis of biomedical image sequences (4D-FAMIS). HeLa and SiHa cells containing, respectively 20-50 and 1-2 copies per cell of human papillomavirus (HPV) DNA type 18 and 16 integrated in cellular DNA were used as models. HPV-DNA was identified using DNA probes containing the whole genome of HPV-DNA type 18 or 16, and DNA-DNA hybrids were revealed by alkaline phosphatase and Fast Red. Cell nuclei were counterstained with thiazole orange (TO) or TOTO-iodide. 4D image sequences were obtained using successive dynamic or spectral sequences of images on different optical sections from CLSM. The location of fluorescent signals within the preparations was determined by FAMIS. This original method summarizes image sequences into a reduced number of images called factor images, and curves called factors. Factors estimate different individual physical behaviours in the sequence such as extinction velocity, spectral patterns and depth emission profiles. Factor images correspond to spatial distributions of the different factors. We distinguished between Fast Red and nucleus stainings in HPV-DNA hybridization signals by taking into account differences in their extinction velocities (fluorescence decay rate) or spectral patterns, and in their focus (depth emission profiles). In HeLa cells, factor images showed that Fast-Red-stained targets could be distinguished from nucleus stainings, and were located on different focal planes of the nuclei. In SiHa cells, 4D-FAMIS determined as few as 1-2 copies per cell of HPV-DNA type 16 located in continuous focal planes. Therefore, 4D-FAMIS, together with CLSM, made the detection and characterization of low copy numbers of genes in whole cells possible.

Cell Nucleus↗

Spatial regularization applied to factor analysis of medical image sequences (FAMIS).

Dynamic image sequences allow physiological mechanisms to be monitored after the injection of a tracer. Factor analysis of medical image sequences (FAMIS) hence creates a synthesis of the information in one image sequence. It estimates a limited number of structures (factor images) assuming that the tracer kinetics (factors) are similar at each point inside the structure. A spatial regularization method for computing factor images (REG-FAMIS) is proposed to remove irregularities due to noise in the original data while preserving discontinuities between structures. REG-FAMIS has been applied to two sets of simulations: (a) dynamic data with Gaussian noise and (b) dynamic studies in emission tomography (PET or SPECT), which respect real tomographic acquisition parameters and noise characteristics. Optimal regularization parameters are estimated in order to minimize the distance between reference images and regularized factor images. Compared with conventional factor images, the root mean square error between regularized images and reference factor images is improved by 3 for the first set of simulations, and by about 1.5 for the second set of simulations. In all cases, regularized factor images are qualitatively and quantitatively improved.

Algorithms↗

Confocal-multilabeling, ultrasensitive TUNEL analysis of DNA breaks in individual cells.

OBJECTIVE: To visualize and localize fragmented DNA strands within apoptotic cells by means of fluorescence using TdT-mediated dUTP-biotin nick end labeling (TUNEL) techniques, laser scanning confocal microscopy (CLSM) and factor analysis of biomedical image sequences (FAMIS). STUDY DESIGN: For this experiment, lymphoid reverted cells were used as a model. Characteristic DNA breaks inside apoptotic cells were detected using TUNEL techniques by a reaction involving tetramethyl rhodamin isothyocyanate (TRITC). The DNA from cell nuclei was counterstained using chromomycin A3 (CA3). The tandem TRITC-CA3 in CLSM was applied to investigate the ability to detect DNA breaks in individual cells using TUNEL techniques and its amplified variants (TUNEL-CARD). FAMIS was applied on dynamic sequences of images of TUNEL preparations and on four-dimensional (4-D) sequences of images of TUNEL-CARD preparations. RESULTS: Distribution and amplitude of fluorescent structures were characterized on dynamic sequences of images. Characterization was improved when FAMIS was applied on 4-D sequences of images, taking into account differences in photobleaching and/or spectrum of TRITC and CA3. CONCLUSION: It is possible to discriminate targets from CA3. FAMIS and TUNEL methods can be used to visualize and localize multiple DNA breaks in lymphoid reverted cells in improved methods of experimentation.

Cell Nucleus↗