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Cardiopulmonary vascular imaging.

Magnetic resonance angiography (MRA) effectively maximizes the flow sensitivity of MR imaging (MRI) to display the body's vasculature. Although MRA has proven quite effective in imaging the intracranial and extracranial vessels, cardiopulmonary MRA faces several additional inherent challenges. Cardiorespiratory motion, vessel pulsatility, and irregular flow patterns all degrade image quality and necessitate appropriate compensation schemes. Although preliminary cardiopulmonary MRA methods appear promising, near instantaneous scan times may be necessary to fully maximize the potential of this technique. Multislice spin-echo and gradient-echo techniques have also proven effective. These methods may compensate for cardiac motion through either ECG-gating or referencing, and may maximize the signal characteristics from the intrinsic slow blood flow of the pulmonary vasculature. Phase velocity mapping techniques, which have proven capable of quantifying blood flow in the body, appear equally promising in preliminary studies in evaluating the cardiopulmonary vascular system. Because of the nature of blood flow dynamics in the cardiopulmonary vascular system, spin-echo (SE), gradient refocussed echo (GRE), and possibly phase mapping technique appear well suited for evaluating these regions.

Artifacts↗

Three-dimensional crystallographic reconstruction for atomic resolution.

Three-dimensional structures have recently been determined by electron crystallography at a resolution high enough to determine atomic arrangements in both protein and mineral specimens. The different nature of these two types of specimens produces some very significant differences in the way data is obtained and processed, although the principles are the same. The sensitivity of proteins to damage by the electron beam limits the signal-to-noise ratio in the image and the resolution to which data can be extracted from the image. A number of constraints, such as the amino acid sequence and the connectivity of atoms within amino acids, can be used in interpreting the limited image data. In materials samples, the relative insensitivity to damage allows obtaining resolution limited only by the microscope. In many samples, dynamical scattering and other non-linear effects limit the information in the image, but this limit can be circumvented by working in very thin areas of the specimen.

Bacteriorhodopsins↗

Noninvasive external recording of cardiac conduction system (His bundle) activity.

Successful and adequate external recording of the cardiac conduction system from the body's surface can be accomplished in 80 to 90 percent of subjects studied. High-gain amplification, signal averaging, and triggering with a conditioned QRS signal results in good recording reproducibility. Averaging of 128 consecutive cycles is adequate, but on occasion averaging of 256 cycles may yield better results. The patients's QRS signal triggers the transfer of signals, which are digitized and stored during the preceding P-R interval. Comparison of external recordings with direct invasive recordings in animals and patients shows good correlation between the major His bundle deflections. The advantages of the system developed include its mobility, triggering the QRS with pretrigger data processing, and instantaneous display on Polaroid photograph. Future research should concentrate on further miniaturization and simplification of the instrumentation, detailed experimental comparison between direct and external recordings for identification of deflections and their origin, further study of the recording lead system, and the most appropriate method of information display.

Animals↗

Alignment, classification, and three-dimensional reconstruction of single particles embedded in ice.

Cryo-electron microscopy of single biological particles poses new challenges to digital image processing due to the low signal-to-noise ratio of the data. New tools have been devised to deal with important aspects of 3-D reconstruction following the random-conical data collection scheme: (a) a new shift-invariant function has been derived, which promises to facilitate alignment and classification of single particle projections; (b) a new method of orientation search is proposed, which makes it possible to relate random-conical data sets to one another prior to reconstruction; and (c) the foundation is laid for a 3-D variance estimation which utilizes the oversampling of 3-D angular space by projections in the random-conical reconstruction scheme.

Algorithms↗

Magnetic resonance neurography.

We have made cross-sectional image "neurograms" in which peripheral nerve has a greater signal intensity than that of other tissue. Neurographic images of the rabbit forelimb were obtained using a spin-echo magnetic resonance imaging (MRI) technique that combines fat suppression and diffusion weighting. After fat suppression the nerve shows up in relative isolation and is brighter than the surrounding tissue due to its longer T2 relaxation time of approximately 50 ms compared to approximately 27 ms for muscle. The addition of pulsed gradients for diffusion weighting of the MR signal further enhances the intensity of the nerve signal relative to that of surrounding muscle tissue. The greater diffusional anisotropy of nerve tissue (D parallel/D perpendicular = 3.1) compared to that of muscle (D parallel/D perpendicular = 1.9) allows further enhancement of the nerve by a subtraction of two diffusion-weighted images, one with the gradients oriented parallel and one with the gradients oriented perpendicular to the nerve orientation. We show that by manipulation of the MRI parameters, either echo time or pulsed gradient strength, the nerves can be made to show up as the most intense feature. This verifies the feasibility of generating three-dimensional "neurographic" images, analogous to angiograms, but which demonstrate the peripheral nerve tracts in apparent isolation.

Adipose Tissue↗

Magnetization-prepared MR angiography with fat suppression and venous saturation.

Magnetization-prepared magnetic resonance (MR) angiography (MPMRA) is an inflow-based two-dimensional (2D) imaging sequence in which a preparation phase precedes rapid image acquisition. For maximal blood/tissue contrast, an inversion-recovery preparation nulls signal from static tissue. If needed, a second inversion suppresses signal from fat. Fully magnetized blood flows in after the inversion pulse(s), providing high signal intensity. The centric phase-encoding order, which ensures that the initial contrast is reflected in the image set, requires the use of a modified venous saturation technique. The sequence is described and its performance assessed with regard to (a) depiction of in-plane flow, (b) fat suppression, and (c) venous saturation. Phantom and volunteer studies showed good performance in all three areas. MPMRA images, acquired in just 2-4 seconds per image, had a blood/tissue contrast-to-noise ratio nearly twice that of standard 2D time-of-flight MR angiograms, acquired in 5-7 seconds. The technique is promising for restless patients and in anatomic areas plagued by motion degradation.

Adipose Tissue↗

Uses and limitations of spoiled gradient-refocused imaging in the evaluation of suspected intracranial tumors.

This article describes the use of a radiofrequency-spoiled gradient-recalled (SPGR) imaging pulse sequence in the evaluation of intracranial masses. This pulse sequence provides excellent anatomic detail with T1-weighted image contrast. Rapid, sequential, single-slice (two-dimensional) images of the brain can be obtained in patients who are unable to hold still for long periods of time. In addition, volumetric (three-dimensional) image data sets can be obtained that provide extremely thin (1- to 2-mm) sections of high detail and good signal-to-noise ratio for selected critical structures within the brain. Finally, because SPGR is also utilized for time-of-flight angiography, parenchymal information can be obtained simultaneously with cerebral blood vessel definition. One potential pitfall that the magnetic resonance radiologist must be aware of is the fact that, in many patients, the degree of contrast enhancement is greatly diminished on postgadolinium SPGR images compared with conventional spin-echo T1-weighted images. Comparison with a set of standard spin-echo postcontrast images or, potentially, the use of higher doses of gadolinium may solve this problem. In spite of this limitation, the selective utilization of SPGR imaging can yield additional useful information for evaluation and preoperative planning in patients with intracranial masses.

Brain Neoplasms↗

The recording and analysis of EMG and jaw tracking. I. The recording procedure.

A computer-based system is described which records and analyses electromyographic (EMG) signals and tracking data for mandibular movements during function. EMG signals were obtained bilaterally from six muscles and a Sirognathograph monitored the position of the lower incisor teeth in three dimensions. Directed jaw movement tasks by subjects were monitored. These data were continuously sampled at a rate of 1 KHz per channel over an operator controlled period. Analysis of the data was performed off-line following the experiment. Individual chewing cycles associated with masticatory function were identified and the timing and dimensions of each chewing envelope established. Corresponding EMG values were also determined and displays of EMG and jaw movement were plotted on a common time base. An average chewing cycle and correlated EMG activity could also be displayed on a high resolution monitor and plotted for a hard copy to enable a comparison to be made between subjects with and without jaw dysfunction.

Analysis of Variance↗

Reconstruction of the electrocardiogram during heart surgery.

Electrocardiograms (ECG) recorded during arrhythmia surgery are used for identification of arrhythmias of different morphology. However, the interpretation of an intraoperative ECG is difficult because some leads cannot be recorded and the signals of the remaining leads often differ from those of a preoperative recording because of the sternotomy. Therefore, a method for reconstruction of a complete intraoperative ECG, which resembles a preoperatively recorded ECG, was studied in 24 patients undergoing heart surgery. The reconstruction method involves calculating coefficients for a transformation matrix, using a preoperative ECG recording and a first intraoperative ECG recording. Once this matrix has been established, further intraoperative recordings can be transformed into an ECG which strongly resembles a preoperative ECG. The correlation between reconstructed intraoperative leads and the corresponding preoperative leads was high in the leads Vx and Vy (median correlation coefficient 0.98 and 0.97) and slightly smaller in lead Vz (0.94). Further studies will prove if the method can be useful in arrhythmia surgery.

Aged↗

Focal liver lesions: MR imaging with Mn-DPDP--initial clinical results in 40 patients.

Manganese (II) N,N'-dipyridoxylethylenediamine-N,N'-diacetate-5,5'-bis(phosphate) (DPDP) was evaluated as a contrast agent for magnetic resonance (MR) imaging (1.5 T) of focal liver lesions in 40 patients. Doses of 5 and 10 mumol/kg were administered intravenously. Mn-DPDP-enhanced T1-weighted images were compared quantitatively and subjectively with standard T1- and T2-weighted nonenhanced images. Use of Mn-DPDP resulted in a statistically significant increase in signal intensity of liver parenchyma in T1-weighted images at both doses. No enhancement was seen in metastases, cholangiocarcinomas, or lymphomas, while all hepatocellular carcinomas were enhanced. Enhancement was seen in focal nodular hyperplasia and in regenerative nodules. The lesion-to-liver contrast in Mn-DPDP-enhanced gradient-recalled-echo images was superior to that of all precontrast images (P less than .01). The number of nonenhancing malignant liver lesions detected in spin-echo (SE) images was increased (272 in T2-weighted SE images vs 390 in T1-weighted Mn-DPDP-enhanced SE images). Image interpretation (eg, visualization and demarcation of the lesions) was markedly better in Mn-DPDP-enhanced images than in all precontrast images (P less than .001).

Adult↗

Dynamic analysis of drug action on in vitro reconstituted thyroid follicle by microinjection of tracer molecules and videomicroscopy.

Thyroid cells isolated from the gland by trypsinization are capable in culture of reconstituting histiotypic structures, the thyroid follicles. This morphological differentiation requires the presence of the main thyroid regulator; thyrotropin. We have analyzed some structural and functional aspects of in vitro reconstituted thyroid follicles (RTF) using microinjection of fluorescent probes and videomicroscopy. This experimental approach allowed to visualize biological processes and actions of drugs, signalling factors, etc. in living cells. We describe here some examples of what can be studied with this powerful still-undervalued method. Microinjection of a cell-impermeant fluorescent probe of either high or low molecular mass into the lumen of RTF allowed to check the tightness of this compartment and therefore to analyze the control of tight junctions assembly. A small cell-impermeant probe like Lucifer Yellow microinjected into a cell was used to demonstrate and then to study the regulation of cell to cell communication via gap junctions. The presence of calcium in the lumen of RTF was detected by microinjection of a properly designed probe: Calcium Green which becomes fluorescent in the presence of the ligand. The lumen to cell transport or endocytosis of thyroglobulin, the thyroid prohormone, which is stored into the lumen of the follicles, is currently studied by microinjection of TRITC-labeled thyroglobulin. Coupled to image processing and videorecorder systems, kinetic analysis and quantitative measurements can be performed.

Animals↗

A novel principle for quantitation of fast intracellular calcium changes using Fura-2 and a modified image processing system--applications in studies of neutrophil motility and phagocytosis.

A new principle is described for imaging intracellular free calcium [Ca2+]i changes in single, living cells utilizing the fluorescent probe Fura-2. It is based upon video color mixing in real time and allows high-speed visualization, at maximum image resolution, of [Ca2+]i changes without digital image ratioing. The epifluorescence images produced by 340 and 380 nm excitations are stored in two memory buffers of a personal computer-based image processing system. Two video signals are generated independently from each buffer and connected to the red and green inputs of a video display. An image is this way created, in which [Ca2+]i shows up as a specific hue, whereas changes in dye concentration, light intensity, cell thickness show up as variations in brightness of the imaged cells. The method has advantages over conventional ratio imaging, notably simplicity and speed, since no calculations are made. Yet it can be combined with traditional digital image processing. The imaging technique allows monitoring of [Ca2+]i changes in rapidly moving cells, like neutrophils. It is demonstrated that during random locomotion on serum-coated glass surfaces, [Ca2+]i levels appeared to oscillate and that the frequency of the oscillations are related to locomotive activity. Furthermore, in Ca2+ free medium, the cells continue to move and phagocytose in the presence of Ca2+ ionophore (ionomycin) and 2 mM EGTA. In the presence of 1 mM extracellular Ca2+, ionomycin-treated cells were not able to move or phagocytose.

Calcium↗

Poly(A) RNA codistribution with microfilaments: evaluation by in situ hybridization and quantitative digital imaging microscopy.

The distribution of poly(A) RNA has been visualized in single cells using high-resolution fluorescent in situ hybridization. Digital imaging microscopy was used to quantitate the signal in various cellular compartments. Most of the poly(A) signal remained associated with the cellular filament systems after solubilization of membranes with Triton, dissociation of ribosomes with puromycin, and digestion of non-poly(A) RNA with ribonuclease A and T1. The actin filaments were shown to be the predominant cellular structural elements associating with the poly(A) because low doses of cytochalasin released about two-thirds of the poly(A). An approach to assess the extent of colocalization of two images was devised using in situ hybridization to poly(A) in combination with probes for ribosomes, membranes, or F-actin. Digital imaging microscopy showed that most poly(A) spatially distributes most significantly with ribosomes, slightly less with F-actin, and least of all with membranes. The results suggest a mechanism for anchoring (and perhaps moving) much of the cellular mRNA utilizing the interaction between actin filaments and poly(A).

Actin Cytoskeleton↗

Interleukin-1 is a motility factor for human breast carcinoma cells in vitro: additive effect with interleukin-6.

Interleukin-1 beta (Il-1 beta) and interleukin-1 alpha (Il-1 alpha) were shown to act as motility factors for the human breast carcinoma cell lines SK-BR-3 and ZR-75-1 in vitro. Both cytokines induced transition from the stationary to the motile phenotype (spreading). Il-1 beta stimulated translocation, shape change and random migration (chemokinesis) of SK-BR-3 cells as demonstrated by time-lapse video recordings and by a modified Boyden chamber assay. Interleukin-6 (Il-6) stimulated spreading of the SK-BR-3 cells; an additive effect with Il-1 beta on spreading and fast plasma membrane movements was evidenced. In the SK-BR-3 cell line, the signal transduction of Il-1 beta and Il-6 differed, since only the effect of Il-6 on spreading was sensitive to pertussis toxin. Both Il-1 beta and Il-6 required protein synthesis to stimulate spreading, since cycloheximide inhibited the effect of the cytokines. Induction of an autocrine loop of Il-6 in the SK-BR-3 cells by Il-1 beta was unlikely, since after stimulation with Il-1 beta, no induction of Il-6 activity was measured, nor was inhibition of stimulated spreading seen in the presence of an antiserum against Il-6. Addition of Il-8 or of an antiserum against Il-8 did not affect spreading. We concluded that Il-1 and Il-6 could act as motility factors for human breast carcinoma cells, in both an independent and an additive way.(ABSTRACT TRUNCATED AT 250 WORDS)

Breast Neoplasms↗

[Clinical potentialities and limitations of two- and three-dimensional time-of-flight MR angiography in the diagnosis of carotid stenosis].

For diagnosing a carotid artery stenosis, 26 patients underwent MR angiography. A spin echo (SE) sequence with presaturation on which the flowing blood appears with low signal intensity was applied, as well as a two- and three-dimensional gradient echo (GRE) sequence with flow compensation, showing blood flow with high signal intensity. Subsequently, projection angiograms were made from the MR images with a maximum intensity projection algorithm. Degree, length and localisation of a carotid artery stenosis were reviewed. To find out the clinical usability, the results of the MR angiograms were compared subsequently with the findings of intravenous digital subtraction angiography (i.v. DSA). In comparison with DSA a correlation in the degree of stenosis was noticed in 42 out of 48 SE-, in 39 out of 52 2-D GRE and in 41 out of 48 3-D GRE sequence angiograms. The length of 34 moderate and severe stenoses, demonstrated by DSA, was overestimated 5 times with the SE-sequence, 9 times with the 3-D and 23 times with the 2-D GRE sequence. MR angiography with a 3-D GRE sequence is suitable for screening for carotid artery stenosis. In cases of severe stenosis an SE sequence should be performed for more precise delineation of the stenotic lesion.

Aged↗

An NMR phased array for human cardiac 31P spectroscopy.

A four-coil phased-array 31P NMR receiver was designed and tested for human cardiac applications, to determine whether the combination of relatively high signal-to-noise ratio (SNR) and large field of view produced in 1H imaging is also realized for in vivo 31P spectroscopy. Spectra were acquired in parallel from an array of four overlapping 6.5-cm surface coils using one- and two-dimensional phase-encoding pulse sequences and were optimally combined to yield composite spectroscopic images. The phased array was found to generate useful 31P spectra from a 2.5-fold wider lateral region around the anterior myocardium than a single receiver of the same size as the array elements, with no increase in imaging time. In addition, the sensitive depth was increased by up to 2 cm over that of a single coil. Spectra could be acquired in roughly 15 min from a region extending to the middle of the heart, with voxel sizes of 2 x 2 x 4 cm3. For the average heart voxel, the SNR of the combined spectrum was higher than that of the best spectrum from any one coil in the array by 30%, with some voxels showing an increase as high as 60%.

Heart↗

Magnetic resonance angiography techniques.

After a radio frequency pulse, the decay of the magnetic resonance (MR) signal is described by two relaxation processes, T1 and T2. T1 describes the rate at which the magnetization realigns itself along the external magnetic field direction (ML), and T2 describes the rate of decay of the magnetization component along the transverse axis (MT). Magnetic resonance angiography (MRA) sequences have been developed that encode flow as changes in the apparent T1 or T2 of the moving blood relative to stationary tissues. MRA sequences typically use either time-of-flight (TOF) techniques to encode T1 or phase-contrast techniques to encode T2. TOF techniques encode flow as an apparent T1 shortening through the wash-in of fully relaxed blood from outside the image volume. The shorter T1 produces an enhancement of vascular structures relative to stationary tissues. TOF methods may use either sequential two-dimensional, three-dimensional, or multi-slab three-dimensional imaging sequences to produce a three-dimensional MRA data set. Phase-contrast methods use additional magnetic field gradients to encode flow as shifts in the phase of MT. Both TOF and phase-contrast methods use maximum intensity projection (MIP) images displayed in a cine format to aid in the visualization of three-dimensional vascular structures.

Animals↗

Quantification of inter- and intra-nuclear variation of fluorescence in situ hybridization signals.

This study aims at the quantification of specific DNA sequences by using fluorescence in situ hybridization (ISH) and digital imaging microscopy. The cytochemical and cytometric aspects of a quantitative ISH procedure were investigated, using human peripheral blood lymphocyte interphase nuclei and probes detecting high copy number target sequences as a model system. These chromosome-specific probes were labeled with biotin, digoxigenin, or fluorescein. Quantification of the fluorescence ISH signals was performed using an epifluorescence microscope equipped with a multi-wavelength illuminator, and a cooled charge coupled device (CCD) camera. Specific image analysis programs were developed for the segmentation and analysis of the images provided by ISH. The fluorescence intensity distributions of the ISH spots showed large internuclear variation (CVs up to 65%) for the probes used. The variation in intensity was found to be independent of the probe, the type of labeling, and the type of immunocytochemical detection used. Variation in intensity was not caused primarily by the immunocytochemical detection method, since directly fluorescein-labeled probes showed similar internuclear variation. Furthermore, it was found that different white blood cell types, which harbor different degrees of compactness of the nuclear chromatin, showed the same variation. The intra-nuclear variation in intensity of the ISH spots on the two chromosome homologs within one nucleus was significantly smaller (approximately 20%) than the inter-nuclear variation, probably due to more constant local hybridization conditions. Due to the relatively small intranuclear variation, copy number polymorphisms of the satellite DNA sequence on chromosome 1 could readily be quantified.(ABSTRACT TRUNCATED AT 250 WORDS)

Analog-Digital Conversion↗