New views on RPE65 deficiency: the rod system is the source of vision in a mouse model of Leber congenital amaurosis.

Leber congenital amaurosis (LCA) is the most serious form of the autosomal recessive childhood-onset retinal dystrophies. Mutations in the gene encoding RPE65, a protein vital for regeneration of the visual pigment rhodopsin in the retinal pigment epithelium, account for 10-15% of LCA cases. Whereas previous studies of RPE65 deficiency in both animal models and patients attributed remaining visual function to cones, we show here that light-evoked retinal responses in fact originate from rods. For this purpose, we selectively impaired either rod or cone function in Rpe65-/- mice by generating double- mutant mice with models of pure cone function (rhodopsin-deficient mice; Rho-/-) and pure rod function (cyclic nucleotide-gated channel alpha3-deficient mice; Cnga3-/-). The electroretinograms (ERGs) of Rpe65-/- and Rpe65-/-Cnga3-/- mice were almost identical, whereas there was no assessable response in Rpe65-/-Rho-/- mice. Thus, we conclude that the rod system is the source of vision in RPE65 deficiency. Furthermore, we found that lack of RPE65 enables rods to mimic cone function by responding under normally cone-isolating lighting conditions. We propose as a mechanism decreased rod sensitivity due to a reduction in rhodopsin content to less than 1%. In general, the dissection of pathophysiological processes in animal models through the introduction of additional, selective mutations is a promising concept in functional genetics.

Diffusion-weighted MRI measurements on stroke patients reveal water-exchange mechanisms in sub-acute ischaemic lesions.

The aim of this study was to investigate the diffusion time dependence of signal-versus-b curves obtained from diffusion-weighted magnetic resonance imaging (DW-MRI) of sub-acute ischaemic lesions in stroke patients. In this case series study, 16 patients with sub-acute ischaemic stroke were examined with DW-MRI using two different diffusion times (60 and 260 ms). Nine of these patients showed sufficiently large lesions without artefacts to merit further analysis. The signal-versus-b curves from the lesions were plotted and analysed using a two-compartment model including compartmental exchange. To validate the model and to aid the interpretation of the estimated model parameters, Monte Carlo simulations were performed. In eight cases, the plotted signal-versus-b curves, obtained from the lesions, showed a signal-curve split-up when data for the two diffusion times were compared, revealing effects of compartmental water exchange. For one of the patients, parametric maps were generated based on the extracted model parameters. These novel observations suggest that water exchange between different water pools is measurable and thus potentially useful for clinical assessment. The information can improve the understanding of the relationship between the DW-MRI signal intensity and the microstructural properties of the lesions.

Functional evaluation of extracardiac ventriculopulmonary conduits and of the right ventricle with magnetic resonance imaging and velocity mapping.

Extracardiac ventriculopulmonary conduits tend to deteriorate over time, developing both obstruction and regurgitation. In this prospective study, magnetic resonance imaging (MRI) was compared with Doppler echocardiography to determine whether MRI improves the noninvasive evaluation of conduit patients. Twenty-five patients (median age 10 years, range 2.5 to 32) were investigated 27 times with Doppler echocardiography and an MRI protocol with spin echo sequences for morphology, velocity mapping, and multislice gradient echo technique for right ventricular volume measuring. Cardiac catheterization data were available in 6 patients. Echocardiography could assess the morphology of the conduits in 6 patients, whereas MRI demonstrated all conduits efficiently. Doppler echocardiography could evaluate the occurrence of regurgitation in 18 patients and could quantify peak velocity in 20 of the patients. A technically adequate MRI velocity mapping was obtained in 25 patients. There was good agreement between MRI and Doppler echocardiography in establishing or not establishing regurgitation, but Doppler echocardiography was less reliable in evaluating the degree of regurgitation. The correlation between peak velocity determined with Doppler and magnetic resonance imaging was r = 0.63 [corrected]. Correlations between catheterization pressure gradients and noninvasive techniques were r = 0.97 for magnetic resonance imaging [corrected] versus catheterization, and r = 0.86 [corrected] for Doppler versus catheterization. MRI can provide complete information on the morphology and function of extracardiac ventriculopulmonary conduits, as well as of the right ventricle. If the results of MRI and echocardiography with Doppler are in agreement, heart catheterization and angiography can be avoided, even in patients considered for conduit replacement.

Gradient echo imaging of flowing hyperpolarized nuclei: theory and phantom studies on 129Xe dissolved in ethanol.

The influence of flip angle and flow velocity on the signal intensity achieved when imaging a hyperpolarized substance with a spoiled gradient echo sequence was investigated. The study was performed both theoretically and experimentally using hyperpolarized xenon dissolved in ethanol. Analytical expressions regarding the optimal flip angle with respect to signal and the corresponding signal level are presented and comparisons with thermally polarized substances are made. Both experimentally and theoretically, the optimal flip angle was found to increase with increasing flow velocity. Numerical calculations showed that the velocity dependence of the signal differs between the cases of hyperpolarized and thermally polarized substances.

Demonstration of 99mTc-labelled plasmin on the surface of ex vivo thrombi.

In an vitro system using the Chandler model for the preparation of in vitro thrombi trace amounts of porcine or human 99mTc-labelled plasmin was found to adsorb to the surface of a preformed thrombus. A radioactive lining of the thrombus could be demonstrated using autoradiography after addition of 99mTc-labelled plasmin in concentrations of 0.04 - 0.07 or 0.4 - 0.7 CTA u/thrombus made from 2 ml whole blood (0.035 - 0.35 microM). The same pattern was found for porcine as for human plasmin. The presence of tranexamic acid in concentrations of 3 to 12 mM did not affect the binding of plasmin indicating that the plasmin binding to fibrin was independent of the lysine binding sites. Furthermore alpha 2-antiplasmin was demonstrated on/in the thrombus also when no plasmin was present indicating a binding of alpha 2-antiplasmin to the thrombus. The plasmin bound to the thrombus was proteolytically inactive. In order to obtain thrombolysis most of the alpha 2-antiplasmin in the surrounding medium had to be neutralized.

Increased brain water self-diffusion in patients with idiopathic intracranial hypertension.

PURPOSE: To investigate changes in brain water diffusion in patients with idiopathic intracranial hypertension. METHODS: A motion-compensated MR pulse sequence was used to create diffusion maps of the apparent diffusion coefficient (ADC) in 12 patients fulfilling conventional diagnostic criteria for idiopathic intracranial hypertension and in 12 healthy volunteers. RESULTS: A significantly larger ADC was found within subcortical white matter in the patient group (mean, 1.16 x 10(-9) m2/s) than in the control group (mean, 0.75 x 10(-9) m2/s), whereas no significant differences were found within cortical gray matter, the basal nuclei, the internal capsule, or the corpus callosum. Four of 7 patients with increased ADC in subcortical white matter also had increased ADC within gray matter. CONCLUSION: Measurement of diffusion coefficients in vivo demonstrated increased local water mobility within subcortical white matter in 7 patients with idiopathic intracranial hypertension that otherwise appeared normal on conventional MR imaging. Further studies are necessary to assess the clinical significance of these observations.

Proton MR spectroscopy and preoperative diagnostic accuracy: an evaluation of intracranial mass lesions characterized by stereotactic biopsy findings.

BACKGROUND AND PURPOSE: MR imaging has made it easier to distinguish among the different types of intracranial mass lesions. Nevertheless, it is sometimes impossible to base a diagnosis solely on clinical and neuroradiologic findings, and, in these cases, biopsy must be performed. The purpose of this study was to evaluate the hypothesis that proton MR spectroscopy is able to improve preoperative diagnostic accuracy in cases of intracranial tumors and may therefore obviate stereotactic biopsy. METHODS: Twenty-six patients with intracranial tumors underwent MR imaging, proton MR spectroscopy, and stereotactic biopsy. MR spectroscopic findings were evaluated for the distribution pattern of pathologic spectra (NAA/Cho ratio < 1) across the lesion and neighboring tissue, for signal ratios in different tumor types, and for their potential to improve preoperative diagnostic accuracy. RESULTS: Gliomas and lymphomas showed pathologic spectra outside the area of contrast enhancement while four nonastrocytic circumscribed tumors (meningioma, pineocytoma, metastasis, and germinoma) showed no pathologic spectra outside the region of enhancement. No significant correlation was found between different tumor types and signal ratios. MR spectroscopy improved diagnostic accuracy by differentiating infiltrative from circumscribed tumors; however, diagnostic accuracy was not improved in terms of differentiating the types of infiltrative or circumscribed lesions. CONCLUSION: MR spectroscopy can improve diagnostic accuracy by differentiating circumscribed brain lesions from histologically infiltrating processes, which may be difficult or impossible solely on the basis of clinical or neuroradiologic findings.

A two-compartment phantom for VOI profile measurements in small-bore 31P MR spectroscopy.

A two-compartment gel phantom for VOI profile measurements in volume-selective 31P spectroscopy in small-bore units is presented. The phantom is cylindrical with two compartments divided by a very thin (30 microm) polyethene film. This thin film permits measurements with a minimum of susceptibility influences from the partition wall. The phantom was used for evaluation of the volume selection method ISIS (image-selected in vivo spectroscopy). The position of the phantom was fixed in the magnet during the measurements, while the volume of interest (VOI) was moved stepwise over the border. The signal from the two compartments was measured for each position and the data were evaluated following differentiation. We have found this phantom suitable for VOI profile measurements of ISIS in small-bore systems. The phantom forms a useful complement to recommended phantoms for small bore-spectroscopy.

MRI thermometry in phantoms by use of the proton resonance frequency shift method: application to interstitial laser thermotherapy.

In this work the temperature dependence of the proton resonance frequency was assessed in agarose gel with a high melting temperature (95 degrees C) and in porcine liver in vitro at temperatures relevant to thermotherapy (25-80 degrees C). Furthermore, an optically tissue-like agarose gel phantom was developed and evaluated for use in MRI. The phantom was used to visualize temperature distributions from a diffusing laser fibre by means of the proton resonance frequency shift method. An approximately linear relationship (0.0085 ppm degrees C(-1)) between proton resonance frequency shift and temperature change was found for agarose gel, whereas deviations from a linear relationship were observed for porcine liver. The optically tissue-like agarose gel allowed reliable MRI temperature monitoring, and the MR relaxation times (T1 and T2) and the optical properties were found to be independently alterable. Temperature distributions around a diffusing laser fibre, during irradiation and subsequent cooling, were assessed with high spatial resolution (voxel size = 4.3 mm3) and with random uncertainties ranging from 0.3 degrees C to 1.4 degrees C (1 SD) with a 40 s scan time.

A rotating phantom for the study of flow effects in MR imaging.

A common type of phantom used for the study of flow effects in MR imaging is the tube phantom, where a liquid passes through a set of tubes placed in the main magnetic field of an MR scanner. Among the disadvantages with this type of phantom are that a distribution of velocities is present in each tube, and that quantifications of flow effects using tube phantoms may be very time-consuming. In this work, we describe the design and the properties of a rotating wheel flow phantom used for quantification of the effects of flow through the imaging plane as well as in the imaging plane. The proposed phantom is constructed as a rotating gel-filled wheel, surrounded by static volumes filled with the same gel, and the evaluation of the information from rotating and static parts is made with a specially designed computer program. The phantom can be used as a plug flow phantom covering simultaneously an interchangeable velocity interval, which at present has the range -52 mm/s, +52 mm/s. It is shown that the phantom gives adequate information on the dependence of pixel content on first-order motion in MR modulus and phase images. Among the fields of application are rapid calibration of MR imaging units for flow determination using phase information, as well as testing of pulse sequence characteristics and verification of theoretical predictions concerning the flow dependence in MR images.

Quantification of complex flow using MR phase imaging--a study of parameters influencing the phase/velocity relation.

In this study, we describe how motion-induced phase angle is affected by different flow models and imaging parameters when using the MR flow phase mapping technique. In a phantom with straight as well as constricted tubes, simulating healthy and stenotic vessels, nonpulsatile flow in the velocity range 0-1 m/sec was maintained. The phase/velocity relation was studied for various degrees of complex flow caused by the constriction, and regions with a breakdown in linearity were determined. Further studies in these regions were made regarding the influence of pulse sequence parameters on the phase/velocity relation. The results showed that in poststenotic areas characterized by so-called separated flow, the phase/velocity relation became nonlinear due to dephasing effects. In regions with fully developed turbulent flow in straight tubes, however, no breakdown in linearity was observed. Parameters seen to have a substantial influence on the phase/velocity relation were first- and second-order velocity encoding and voxel size. Finally, a pilot in vivo demonstration of complex flow was done using a sequence designed to be robust with respect to linearity of the phase/velocity relation. The results indicate that the MR phase mapping technique can be used to measure flow quantitatively in regions with complex flow. This opens possibilities for future clinical use of the technique in the study of areas of complex flow such as valvular heart disease.

A theoretical study of amplitude modulation and time shifting in quantitative MR measurements of motion in brain tissue.

MR imaging pulse sequences can be made sensitive to motion by adding gradients with different strengths at different time intervals. In the well-known phase mapping method, such velocity encoding gradients are used to obtain phase information linear to the velocity of the studied object in the direction of the gradient. When very low velocities are studied, a long duration velocity-encoded gradient is required to obtain sufficient velocity sensitivity. In such cases, variation in the object velocity during the execution of the sequence may hamper the accuracy of the method. In this study, we have made a computer simulation of the performance of a phase mapping method sequence (TE = 46 msec) designed for quantitative studies of motion in brain tissue. Using a Gaussian-shaped velocity input function, the time shifting and the amplitude modulation properties of the sequence was studied for various values of the duration, defined as the full width of tenth of maximum (FWTM), of the input function. The time shift corresponded well to the center of the 180 degrees RF pulse, and the amplitude modulation was seen to decrease with increasing time duration of the velocity input function. Applied on in vivo data, where an approximately gaussian-shaped brain motion velocity pattern was assumed to have a duration of 150 msec, the amplitude modulation of the sequence was estimated to 2%.

Pulse sequence design for MR velocity mapping of complex flow: notes on the necessity of low echo times.

Lowering of the echo time (TE) has been proposed as a way to reduce effects of phase dispersion in MR velocity mapping, because a low TE reduces sensitivity to higher-order motion terms while first-order velocity sensitivity is maintained. Methods of lowering TE involves the use of extreme gradient ramp times and gradient strengths as well as reduction of the duration of transmit/receive windows, the latter method causing decrements in image resolution. When reducing higher-order sensitivity, however, it is not the overall TE that is the critical parameter, but rather the time pattern of the gradients used in the experiment. Hence, changes in TE without subsequent variations in gradient pattern would, according to theory, not affect quantitative measurements of complex flow and vice versa. In this study, we experimentally demonstrate this relation and utilize the experience to create a sequence robust towards complex flow without sacrifices in image resolution. Our experimental observations show that variations in TE alone while maintaining the time course of the velocity-encoding gradient does not significantly affect measurements of through-plane average complex flow in the studied velocity range. A parameter that cannot be measured as accurately if TE is increased is the peak flow. A phase mapping sequence with prolonged TE from 3 ms to 5 ms but with short duration of the velocity-encoding (section-selective) gradient and improved in-plane resolution was demonstrated in vivo.

Quantitative diffusion coefficient maps using fast spin-echo MRI.

In this work, we have evaluated the performance of a diffusion-sensitive fast spin-echo (FSE) pulse sequence. The proposed pulse sequence utilises velocity-compensating diffusion-encoding gradients and includes the collection of navigator echoes. Spoiler gradients were inserted in the slice-selecting direction to minimise effects from stimulated echoes. Calculations of the b values showed that cross-terms between imaging gradients and diffusion gradients only led to a marginal increase of b values. Pixel-wise calculation of apparent diffusion coefficient (ADC) maps was performed numerically, considering cross-terms between diffusion-encoding and imaging gradients. The sequences investigated used echo train lengths of 16, 8 and 4 echoes and were encoded in either the slice-, frequency- or phase-encoding direction. In order to allow for higher b values a pulse-sequence version using non-motion compensating diffusion-encoding gradients was written. Phantom measurements were performed and the diffusion coefficients of water and acetone were reasonable. Seven healthy volunteers (age 28-50 years) were examined and apparent diffusion coefficient values agreed well with expected values. Diffusion-weighted images, apparent diffusion coefficient maps and images corresponding to the trace of the diffusion tensor of good quality were retrieved in vivo.

Quality assessment of localization technique performance in small volume in vivo 1H MR spectroscopy.

A new phantom and evaluation method for experimental evaluation of 1H-magnetic resonance spectroscopy single volume localization techniques regarding signal contamination (C), defined as the part of the signal originating outside the volume of interest, is presented. The quality assessment method is based on a spherical phantom with an oil/water interface in order to reduce susceptibility effects, and applied for stimulated-echo acquisition method (STEAM) and spin-echo (SE) sequences, echo times of 270, 135, and 10 ms, and cubic volumes of interest (VOI) of 1(3), 1.5(3), 2(3), 2.5(3), and 3(3) cm3. To be able to mimic measurements of the contamination in three dimensions the physical gradients representing the three orthogonal directions for slice selection were shifted in the pulse sequences. Contamination values in one dimension differed between 6.5% and 8.4% in SE sequences, and between 0.7% and 13.8% in STEAM sequences. In STEAM sequences a decrease of C with increasing VOI size was observed while SE sequences showed comparable C values for the different VOI sizes tested. The total contamination in three dimensions were 19% and 18% in SE and STEAM sequences with a TE of 270 ms, and 7% in a STEAM sequence with a TE of 10 ms, respectively. The presented evaluation method is easily applied to the new phantom and showed high reproducibility.

High-resolution diffusion imaging using phase-corrected segmented echo-planar imaging.

Diffusion magnetic resonance imaging (MRI) was performed with a high-resolution segmented echo-planar imaging technique, which provided images with substantially less susceptibility artifacts than images obtained with single-shot echo-planar imaging (EPI). Diffusion imaging performed with any multishot pulse sequence is inherently sensitive to motion artifacts and in order to reduce motion artifacts, the presented method utilizes navigator echo phase corrections, performed after a one-dimensional Fourier transform along the frequency-encoding direction. Navigator echo phases were fitted to a straight line prior to phase correction to avoid errors from internal motion. In vivo imaging was performed using electro cardiographic (ECG) triggering. Apparent diffusion coefficient (ADC) maps were calculated on a pixel-by-pixel basis using up to seven diffusion sensitivities, ranging from b = 0 to 1129 x 10(6) s/m(2).

Quantification of low-velocity motion using a navigator-echo supported MR velocity-mapping technique: application to intracranial dynamics in volunteers and patients with brain tumours.

Gradient-echo pulse sequences with velocity-encoding gradients of 22.5-25 mT/m, were used for brain-motion and CSF-flow studies. To reduce motion artifacts, a phase-correction technique based on navigator echoes was evaluated. Three patients with right-sided parietal tumours were investigated; one astrocytoma grade III-IV, one astrocytoma grade I-II and one benign meningioma. In healthy volunteers, a maximal brain-tissue velocity of (0.94 +/- 0.26) mm/s (mean +/- 1SD) was observed, which is consistent with previously presented results. The phase correction was proven useful for reduction of artifacts due to external head movements in modulus and phase images, without loss of phase information related to internal motion. The tissue velocity within the astrocytomas was low during the entire cardiac cycle. An abnormally high rostral velocity component was, however, observed in the brain tissue frontal to the astrocytomas. In all patients, an abnormal CSF flow pattern was observed. The study of brain motion may provide further understanding of the effects of tumours and other pathological conditions in the brain. When considering intracranial motion as a source of error in diffusion/perfusion MRI, the present study suggests that a pathology can alter the properties of brain motion and CSF flow considerably, leading to a more complex impact on diffusion/perfusion images.

Effects of echo time variation on perfusion assessment using dynamic susceptibility contrast MR imaging at 3 tesla.

The implications of changing the echo time of a gradient-echo echo planar imaging sequence applied to dynamic susceptibility contrast magnetic resonance imaging (DSC-MRI) for perfusion imaging at 3T were investigated. Four echo times in the range of 21 to 45 ms were examined in a total of 17 patients who received a dose of 0.1 mmol/kg bodyweight Gadobutrol (Gadovist, 1.0 mmol/ml). As the primary optimization parameter, the concentration-to-noise ratio (SNRc) was selected as it takes effects of variations in baseline as well as in signal drop into account. In an analysis of gray matter, white matter and arterial regions of interest, SNRc showed the highest values for the shortest applied echo time in all cases. Maps of regional cerebral blood volume (rCBV) and blood flow (rCBF) were calculated using deconvolution based on singular value decomposition. The quality of rCBF and rCBV images was judged to be good or excellent in all cases, independent of the echo time. Calculated gray matter/white matter ratios of rCBF and rCBV displayed no significant dependence on the applied echo time. Considering the better SNRc and arterial signal saturation aspects, we found that the shortest investigated echo time was the superior one. We thus suggest that short echo times should be applied, taking technical limitations and clinical demands into consideration.

A method for MR quantification of flow velocities in blood and CSF using interleaved gradient-echo pulse sequences.

The aim of this study was to establish a rapid method for in vivo quantification of a large range of flow velocities using phase information. A basic gradient-echo sequence was constructed, in which flow was encoded along the slice selection direction by variation of the amplitude of a bipolar gradient without changes in sequence timings. The influence of field inhomogeneities and eddy currents was studied in a 1.5 T interleaved sequences for calibration and in vivo flow determination were constructed, and flow information was obtained by pairwise subtraction of velocity-encoded from velocity non-encoded phase images. Calibration was performed in a nongated mode using flow phantoms, and the results were compared with theoretically calculated encoding efficiencies. In vivo flow was studied in healthy volunteers in three different areas using cardiac gating; central blood flow in the great thoracic vessels, peripheral blood flow in the popliteal vessels, and flow of cerebrospinal fluid (CSF) in the cerebral aqueduct. The results show good agreement with results obtained with other techniques. The proposed method for flow determination was shown to be rapid and flexible, and we thus conclude that it seems well suited for routine clinical MR examinations.

Comparison between retrospective gating and ECG triggering in magnetic resonance velocity mapping.

ECG-triggered cinematographic studies of the cardiovascular system are hampered by several technical restrictions such as the inability to image end-diastole, ghosting, varying signal intensity, and phase contributions from eddy currents. Retrospective gating may solve these problems, but involves signal manipulation such as interpolating raw data from a time window. In this study, the performance of the two gating strategies was compared in quantitative MR velocity mapping on the abdominal aorta in eight healthy volunteers and on a pulsatile flow phantom. The results were compared to a one-dimensional velocity mapping technique and Doppler ultrasound. Finally, the consequence of decreasing the time window in the raw data interpolation used for retrospective gating was also examined. With retrospective gating, a low-pass filtering was seen, causing significantly prolonged duration and decreased amplitude of flow pulses. However, by reducing the time window retrospectively gated flow measurements were in good agreement with those that are ECG triggered. When fulfilling the demand of a narrow time window for interpolation, retrospective gating offers several advantages in MR velocity mapping.