Search

Objectives It has been proposed that successful left ventricular surgical restoration should restore normal helical myofiber orientation. A magnetic resonance imaging technique, magnetic resonance diffusion tensor imaging, has been developed to measure myocyte orientation. By using magnetic resonance diffusion tensor imaging, this study tested the hypothesis that (1) myocyte orientation is altered after anteroapical myocardial infarction and (2) left ventricular surgical restoration restores normal helix angles. Methods Thirteen sheep underwent anteroapical myocardial infarction (25% of left ventricular mass). Ten weeks later, animals underwent either aneurysm plication (n = 8) or sham operations (n = 5). Six weeks after this operation, hearts were excised, perfusion fixed in diastole, and underwent magnetic resonance diffusion tensor imaging. Hearts from normal sheep (n = 5) were also harvested and imaged. Primary eigenvectors of the diffusion tensors from magnetic resonance diffusion tensor imaging were resolved into helix angles relative to a local wall coordinate system. Transmural samples of the helix angles were compared at the border zone of the aneurysm or repair (or a comparable distance from the base in normal sheep), 1 cm below the valves, and halfway between. Results The helical myofiber orientation did not change after myocardial infarction. However, aneurysm plication caused myofibers in the anterior border zone to rotate counterclockwise (−35.6 ± 10.5°, P = .028) and those in the lateral border zone to rotate clockwise (34.4 ± 8.1°, P = .031). Conclusions Surgical restoration alters myocyte orientation adjacent to the surgical repair. However, myofiber orientation is not abnormal after myocardial infarction, and thus surgical restoration techniques intent on restoring normal helix angles might not be warranted.

Although promising for visualizing the structure of ordered tissues, MR diffusion tensor imaging (DTI) has been hampered by long acquisition time and low spatial resolution associated with its inherently low signal-to-noise ratio (SNR). Moreover, the uncertainty in the DTI measurements has a direct impact on the accuracy of structural renderings such as fiber streamline tracking. Noise removal techniques can be used to improve the SNR of DTI without requiring additional acquisitions, albeit most low-pass filtering methods are accompanied by undesirable image blurring. In the present study, a modified vector-based partial-differential-equation (PDE) filtering formalism was implemented for smoothing DTI vector fields. Using an image residual-energy criterion to equate the degree of smoothing and error metrics empirically derived from DTI data to quantify the relative performances, the effectiveness in denoising DTI data is compared among image-based and vector-based PDE and fixed and adaptive low-pass k-space filtering. The results demonstrate that the edge-preservation feature of the PDE approach can be highly advantageous in enhancing DTI measurements, particularly for vector-based PDE filtering in applications relying on DTI directional information. These findings suggest a potential role for the postprocessing enhancement technique to improve the practical utility of DTI.

Diffusion tensor imaging is a variation of magnetic resonance imaging that measures the diffusion of water in tissues. This can help measure and quantify a tissue's orientation and structure, making it an ideal tool for examining cerebral white matter and neural fiber tracts. It is only beginning to be utilized in psychiatric research. This article reviews the theory behind diffusion tensor imaging, its potential to map fiber tracts in the brain, and its recent use in psychiatric research.

The relative utility of 3D, microscopic resolution assessments of fixed mouse myocardial structure via diffusion tensor imaging is demonstrated in this study. Isotropic 100-m resolution fiber orientation mapping within 5.5° accuracy was achieved in 9.1 hr scan time. Preliminary characterization of the diffusion tensor primary eigenvector reveals a smooth and largely linear angular rotation across the left ventricular wall. Moreover, a higher level of structural hierarchy is evident from the organized secondary and tertiary eigenvector fields. These findings are consistent with the known myocardial fiber and laminar structures reported in the literature and suggest an essential role of diffusion tensor microscopy in developing quantitative atlases for studying the structure–function relationships of mouse hearts. Magn Reson Med 52:453– 460, 2004. © 2004 Wiley-Liss, Inc.

MR diffusion tensor imaging (DTI) is a promising tool for characterizing the microstructure of ordered tissues. However, its practical applications are hampered by relatively low signal-to-noise-ratio and spatial and temporal resolution. Reduced-encoding imaging (REI) via k-space sharing with constrained reconstruction has previously been shown to be effective for accelerating DTI, although the implementation was based on rectilinear k-space sampling. Due to the intrinsic oversampling of central k-space and allowance for isotropic downsampling, projection-reconstruction (PR) imaging may be better suited for REI. In this study, regularization procedures, including radial filtering and baseline signal correction to adequately reconstruct reduced encoded PR imaging data, are investigated. The proposed filtered reduced-encoding projection-reconstruction (FREPR) technique is applied to DTI tissue fiber orientation and fractional anisotropy (FA) measurements. Results show that FREPR offers improved reconstructions of the reduced encoded images and on an equal total scan-time basis provides more accurate fiber orientation and FA measurements compared to rectilinear k-space sampling-based REI methods or a control experiment consisting of only fully encoded images. These findings suggest a potentially significant role of FREPR in accelerating repeated imaging and improving the data acquisition-time efficiency of DTI experiments.

Detailed mechanical information of the vein is important to better understand remodeling of the vessel in disease states, but has been difficult to obtain due to its thinness, unique geometry, and limitations of mechanical testing. This study presents a novel method for characterizing deformation of the intact explanted vein under physiological loads and determining its material properties by combining high-resolution imaging and computational analysis. High-resolution CT (microCT) was used to image an iodine-stained, excised porcine internal jugular vein sample under extension to 100 and 120% of in situ length, and inflation and 2, 10, 20 mmHg of pressure, inside a microCT-compatible hydrostatic loading chamber. Regional strains were measured with the finite element (FE) image registration method known as Hyperelastic Warping. Material properties were approximated with inverse FE characterization by optimizing stiffness-related coefficients so to match simulated strains to the experimental measurements. The observed morphology and regional strain of the vein were found to be relatively heterogeneous. The regional variability in the measured strain was primarily driven by geometry. Although iodine treatment may result in tissue stiffening, which requires additional investigation, it is effective in allowing detailed detection of vein geometry. The feasibility and utility of using microCT and computational analysis to characterize mechanical responses and material properties of the vein were demonstrated. The presented method is a promising alternative or addition to mechanical testing for characterizing veins or other similarly delicate vessels in their native anatomical configuration under a wide range of realistic or simulated environmental and loading conditions.

A novel multi-coaxial bioreactor for three-dimensional cultures of adherent cell types, such as liver, is described. It is composed of four tubes of increasing diameter placed one inside the other, creating four spatially isolated compartments. Liver acinar structure and physiological parameters are mimicked by sandwiching cells in the space between the two innermost semi-permeable tubes, or hollows fibers, and creating a radial flow of media from an outer compartment (ECC), through the cell mass compartment, and to an inner compartment (ICC). The outermost compartment is created by gas-permeable tubing, and the housing is used to oxygenate the perfusion media to periportal levels in the ECC. Experiments were performed using distilled water to correlate the radial flow rate (Q(r)) with (1) the pressure drop (DeltaP) between the media compartments that sandwich the cell compartment and (2) the pressure in the cell compartment (P(c)). These results were compared with the theoretical profile calculated based on the hydraulic permeability of the two innermost fibers. Phase-contrast velocity-encoded magnetic resonance imaging was used to visualize directly the axial velocities inside the bioreactor and confirm the assumptions of laminar flow and zero axial velocity at the boundaries of each compartment in the bioreactor. Axial flow rates were calculated from the magnetic resonance imaging results and were similar to the measured axial flow rates for the previously described experiments.

Morphologically accurate biomechanical models of the intervertebral disc anulus fibrosus (AF) require precise knowledge of its lamellar architecture; however, available methods of assessment are limited by poor spatial resolution or the destructive nature of the technique. In a novel approach, diffusion tensor microscopy was used in this study to characterize the microstructure of excised porcine AF samples. Results show diffusion in the AF to be anisotropic. The orientations of anisotropy exhibit a layered morphology that agrees with light micrographs of the corresponding samples, and the behavior of the orientation angles is consistent with the known AF collagen fiber architecture. A static magnetic field-dependent relaxation anisotropy was observed in the AF, which has methodological implications for magnetic resonance (MR) imaging of ordered collageneous tissues. These findings present MR diffusion tensor microscopy as a potentially valuable tool to assess quantitatively and nondestructively water diffusion anisotropy and lamellar structure of the intervertebral disc AF. Magn Reson Med 41:992–999, 1999. © 1999 Wiley-Liss, Inc.

Adiabatic pulses, although useful in generating uniform spin nutation in the presence of inhomogeneous B1 fields, are limited for NMR imaging applications due to the lack of slice-selective excitation capability. Selective excitation techniques using gradient modulation have been introduced; however, present methods require either a minimum of two excitations or eight adiabatic segments. Here, a scheme is presented that allows single-shot, arbitrary flip-angle, and slice-selective excitation with only four adiabatic half-passage segments. The technique is demonstrated via computer simulation and experimental tests on a phantom. Furthermore, issues associated with the implementation of these gradient-modulated adiabatic pulses are discussed.

Functional properties of the myocardium are mediated by the tissue structure. Consequently, proper physiological studies and modeling necessitate a precise knowledge of the fiber orientation. Magnetic resonance (MR) diffusion tensor imaging techniques have been used as a nondestructive means to characterize tissue fiber structure; however, the descriptions so far have been mostly qualitative. This study presents a direct, quantitative comparison of high-resolution MR fiber mapping and histology measurements in a block of excised canine myocardium. Results show an excellent correspondence of the measured fiber angles not only on a point-by-point basis (average difference of −2.30 ± 0.98°, n = 239) but also in the transmural rotation of the helix angles (average correlation coefficient of 0.942 ± 0.008 with average false-positive probability of 0.004 ± 0.001, n = 24). These data strongly support the hypothesis that the eigenvector of the largest MR diffusion tensor eigenvalue coincides with the orientation of the local myocardial fibers and underscore the potential of MR imaging as a noninvasive, three-dimensional modality to characterize tissue fiber architecture.

Non-invasive imaging techniques are highly desirable as an alternative to conventional biopsy for the characterization of the remodeling of tissues associated with disease progression, including end-stage heart failure. Cardiac diffusion tensor imaging (DTI) has become an established method for the characterization of myocardial microstructure. However, the relationships between diffuse myocardial fibrosis, which is a key biomarker for staging and treatment planning of the failing heart, and measured DTI parameters have yet to be investigated systematically. In this study, DTI was performed on left ventricular specimens collected from patients with chronic end-stage heart failure as a result of idiopathic dilated cardiomyopathy (n = 14) and from normal donors (n = 5). Scalar DTI parameters, including fractional anisotropy (FA) and mean (MD), primary (D1 ), secondary (D2 ) and tertiary (D3 ) diffusivities, were correlated with collagen content measured by digital microscopy. Compared with hearts from normal subjects, the FA in failing hearts decreased by 22%, whereas the MD, D2 and D3 increased by 12%, 14% and 24%, respectively (P0.01). No significant change was detected for D1 between the two groups. Furthermore, significant correlation was observed between the DTI scalar indices and quantitative histological measurements of collagen (i.e. fibrosis). Pearson's correlation coefficients (r) between collagen content and FA, MD, D2 and D3 were -0.51, 0.59, 0.56 and 0.62 (P0.05), respectively. The correlation between D1 and collagen content was not significant (r = 0.46, P = 0.05). Computational modeling analysis indicated that the behaviors of the DTI parameters as a function of the degree of fibrosis were well explained by compartmental exchange between myocardial and collagenous tissues. Combined, these findings suggest that scalar DTI parameters can be used as metrics for the non-invasive assessment of diffuse fibrosis in failing hearts.

Displacement fields are important to analyze deformation, which is associated with functional and material tissue properties often used as indicators of health. Magnetic resonance imaging (MRI) techniques like DENSE and image registration methods like Hyperelastic Warping have been used to produce pixel-level deformation fields that are desirable in high-resolution analysis. However, DENSE can be complicated by challenges associated with image phase unwrapping, in particular offset determination. On the other hand, Hyperelastic Warping can be hampered by low local image contrast. The current work proposes a novel approach for measuring tissue displacement with both DENSE and Hyperelastic Warping, incorporating physically accurate displacements obtained by the latter to improve phase characterization in DENSE. The validity of the proposed technique is demonstrated using numerical and physical phantoms, and in vivo small animal cardiac MRI.

Teleost embryos have not been successfully cryopreserved. To formulate successful cryopreservation protocols, the distribution and cellular permeability to water must be understood. In this paper, the zebrafish (Brachydanio rerio) was used as a model for basic studies of the distribution to permeability to water. These embryos are a complex multi-compartmental system composed of two membrane-limited compartments, a large yolk (surrounded by the yolk syncytial layer) and differentiating blastoderm cells (each surrounded by a plasma membrane). Due to the complexity of this system, a variety of techniques, including magnetic resonance microscopy and electron spin resonance, was used to measure the water in these compartments. Cellular water was distributed unequally in each compartment. At the 6-somite stage, the percent water (V/V) was distributed as follows: total in embryo = 74%, total in yolk = 42%, and total in blastoderm = 82%. A one-compartment model was used to analyze kinetic, osmotic shrinkage data and determine a phenomenological water permeability parameter, Lp, assuming intracellular isosmotic compartments of either 40 or 300 mosm. This analysis revealed that the membrane permeability changed (P < 0.05) during development. During the 75% epiboly to 3-somite stage, the mean membrane permeability remained constant (Lp = 0.022 ± 0.002 μm x min-1atm-1 [mean ± S.E.M.] assuming isosmotic is 40 mosm or Lp = 0.049 ± 0.008 μm x min-1atm-1 assuming isosmotic is 300 mosm). However, at the 6-somite stage, Lp increased twofold (Lp = 0.040 ± 0.004 μm x min-1atm-1 assuming isosmotic is 40 mosm or Lp = 0.100 ± 0.017 μm x min-1atm-1 assuming isosmotic is 300 mosm). Therefore, the low permeability of the zebrafish embryo coupled with its large size (and consequent low area to volume ratio) led to a very slow osmotic response that should be considered before formulating cryopreservation protocols. J. Exp. Zool. 278:356–371, 1997. © 1997 Wiley-Liss, Inc.

This paper describes some new approaches for understanding the permeability of teleost embryos. The dechorionated zebrafish (Brachydanio rerio) was used as a model for basic studies of water and cryoprotectant permeability. These embryos are composed of two compartments, a large yolk (surrounded by the yolk syncytial layer) and differentiating blastoderm cells. Cellular water was distributed unequally in each compartment. Measurements indicated that the total water in the embryo was 74%, while the total water in the yolk was 42%, and total water in the blastoderm was 82%. The internal isosmotic value for the zebrafish embryo is unknown. However, for one-compartment modeling studies of membrane permeability, the mean Lp (+/- SEM) values were 0.022 +/- 0.002 to 0.049 +/- 0.008 microns x min-1 atm-1 at 40 mOsm (assuming this was one possible internal isosmotic value for the entire embryo) and 0.040 +/- 0.004 to 0.1 +/- 0.017 microns x min-1 atm-1 at 300 mOsm (assuming this was another possible internal isosmotic value for the entire embryo). When three- and six-somite embryos were placed in 1.5 and 2.0 M cryoprotectants (dimethyl sulfoxide and propylene glycol), osmometric measurements of volume changes indicated no cryoprotectant permeation. However, similar measurements with methanol revealed a small volume decrease (ca. 8%) and recovery (ca. 5%) for six-somite embryos in a 2.0 M solution. Magnetic resonance (MR) images of the spatial distribution of three cryoprotectants (dimethyl sulfoxide, propylene glycol, and methanol) demonstrated that only methanol permeated the entire embryo within 15 min. The other cryoprotectants exhibited little or no permeation into the yolk over 2.5 h. The results from MR spectroscopy and cryoprotectant microinjections into the yolk suggested that the yolk syncytial layer plays the critical limiting role for cryoprotectant permeation throughout the embryo.

Successful cryopreservation of most multicompartmental biological systems has not been achieved. One prerequisite for success is quantitative information on cryoprotectant permeation into and amongst the compartments. This report describes direct measurements of cryoprotectant permeation into a multicompartmental system using chemical shift selective magnetic resonance (MR) microscopy and MR spectroscopy. We used the developing zebrafish embryo as a model for studying these complex systems because these embryos are composed of two membrane-limited compartments: (i) a large yolk (surrounded by the yolk syncytial layer) and (ii) differentiating blastoderm cells (each surrounded by a plasma membrane). MR images of the spatial distribution of three cryoprotectants (dimethyl sulfoxide, propylene glycol, and methanol) demonstrated that methanol permeated the entire embryo within 15 min. In contrast, the other cryoprotectants exhibited little or no permeation over 2.5 h. MR spectroscopy and microinjections of cryoprotectants into the yolk inferred that the yolk syncytial layer plays a critical role in limiting the permeation of some cryoprotectants throughout the embryo. This study demonstrates the power of MR technology combined with micromanipulation for elucidating key physiological factors in cryobiology.

Glioblastoma multiforme, a World Health Organization grade IV glioma, has a poor prognosis in humans despite current treatment options. Here, we present magnetic resonance imaging (MRI) data regarding the regression of aggressive rat F98 gliomas and human U87 glioma xenografts after treatment with the nitrone compound OKN-007, a disulfonyl derivative of α-phenyl-tert-butyl nitrone.MRI was used to assess tumor volumes in F98 and U87 gliomas, and bioluminescence imaging was used to measure tumor volumes in F98 gliomas encoded with the luciferase gene (F98(luc)). Immunohistochemistry was used to assess angiogenesis (vascular endothelial growth factor [VEGF] and microvessel density [MVD]), cell differentiation (carbonic anhydrase IX [CA-IX]), hypoxia (hypoxia-inducible factor-1α [HIF-1α]), cell proliferation (glucose transporter 1 [Glut-1] and MIB-1), proliferation index, and apoptosis (cleaved caspase 3) markers in F98 gliomas. VEGF, CA-IX, Glut-1, HIF-1α, and cleaved caspase 3 were assessed in U87 gliomas.Animal survival was found to be significantly increased (P.001 for F98, P.01 for U87) in the group that received OKN-007 treatment compared with the untreated groups. After MRI detection of F98 gliomas, OKN-007, administered orally, was found to decrease tumor growth (P.05). U87 glioma volumes were found to significantly decrease (P.05) after OKN-007 treatment, compared with untreated animals. OKN-007 administration resulted in significant decreases in tumor hypoxia (HIF-1α [P.05] in both F98 and U87), angiogenesis (MVD [P.05], but not VEGF, in F98 or U87), and cell proliferation (Glut-1 [P.05 in F98, P.01 in U87] and MIB-1 [P.01] in F98) and caused a significant increase in apoptosis (cleaved caspase 3 [P.001 in F98, P.05 in U87]), compared with untreated animals.OKN-007 may be considered as a promising therapeutic addition or alternative for the treatment of aggressive human gliomas.

Cell therapies are potential alternatives to organ transplantation for liver failure or dysfunction but are compromised by inefficient engraftment, cell dispersal to ectopic sites, and emboli formation. Grafting strategies have been devised for transplantation of human hepatic stem cells (hHpSCs) embedded into a mix of soluble signals and extracellular matrix biomaterials (hyaluronans, type III collagen, laminin) found in stem cell niches. The hHpSCs maintain a stable stem cell phenotype under the graft conditions. The grafts were transplanted into the livers of immunocompromised murine hosts with and without carbon tetrachloride treatment to assess the effects of quiescent versus injured liver conditions. Grafted cells remained localized to the livers, resulting in a larger bolus of engrafted cells in the host livers under quiescent conditions and with potential for more rapid expansion under injured liver conditions. By contrast, transplantation by direct injection or via a vascular route resulted in inefficient engraftment and cell dispersal to ectopic sites. Transplantation by grafting is proposed as a preferred strategy for cell therapies for solid organs such as the liver. (HEPATOLOGY 2013)

A modified spin-echo pulse sequence is described that enables accurate T2 measurements to be made in NMR microimaging experiments. The modified sequence eliminates cumulative diffusion losses that lead to an underestimation of the T2 relaxation time using conventional spin-echo pulse sequences. The approach is theoretically justified and confirmed in comparative experiments on phantoms.

NMR microimages of single neural cells were acquired at 500 MHz using a conventional spin echo pulse sequence and a line-narrowing sequence that eliminates susceptibility effects. The data show that any contribution to the measured T2 relaxation rate arising from diffusion in local field inhomogeneities using spin echo sequences at high fields and high spatial resolution is relatively small. We conclude that the measured T2 difference between the nucleus and cytoplasm in these cells represents primarily a true T2 relaxation effect arising from the interactions of water with macromolecules in the two compartments and does not result from microsusceptibility differences. These observations have implications regarding water compartmentation in single cells and the interpretation of the MR characteristics of tissues in vivo.

Relaxation-time and diffusion-weighted NMR micrographs have been obtained for single neurons isolated from Aplysia californica. These images allow the nucleus and cytoplasm to be clearly differentiated, in contrast to proton spin-density images, which appear relatively homogenous. Images of the spatial distribution of T1 and T2 relaxivities and the diffusion coefficient (D), as well as average values for T1, T2, and D in the cytoplasm and nucleus, were calculated from sets of appropriately weighted images. In all cases, water in the nucleus had relaxation and diffusion properties markedly differing from those of cytoplasmic water, which in turn had properties which were distinct from those of free water. Additionally, the cytoplasmic T2 was observed to triple following cell death, which is attributed to cytoplasmic dilution as water enters the cell. The work presented represents the first effort at a consistent exploration of the spatial distribution of NMR characteristics of water within intact single cells. These studies have implications both for modeling the NMR characteristics of water in neuronal tissues based on an understanding of the characteristics of water in different cell compartments and for understanding water/macromolecule interactions within cells. NMR microscopy studies such as these may help form a foundation for understanding and interpreting NMR characteristics measured from large assemblies of cells, i.e., spectroscopy and imaging of living tissues.

Background Accurate interpretations of cardiac functions require precise structural models of the myocardium, but the latter is not available always and for all species. Although scaling or substitution of myocardial fiber information from alternate species has been used in cardiac functional modeling, the validity of such practice has not been tested. Methods Fixed mouse (n = 10), rabbit (n = 6), and sheep (n = 5) hearts underwent diffusion tensor imaging (DTI). The myocardial structures in terms of the left ventricular fiber orientation helix angle index were quantitatively compared between the mouse rabbit and sheep hearts. Results The results show that significant fiber structural differences exist between any two of the three species. Specifically, the subepicardial fiber orientation, and the transmural range and linearity of fiber helix angles are significantly different between the mouse and either rabbit or sheep. Additionally, a significant difference was found between the transmural helix angle range between the rabbit and sheep. Across different circumferential regions of the heart, the fiber orientation was not found to be significantly different. Conclusions The current study indicates that myocardial structural differences exist between different size hearts. An immediate implication of the present findings for myocardial structural or functional modeling studies is that caution must be exercised when extrapolating myocardial structures from one species to another.

In this paper, we propose three metrics to quantify the differences between the results of diffusion tensor magnetic resonance imaging (DT-MRI) fiber tracking algorithms: the area between corresponding fibers of each bundle, the Earth Mover's Distance (EMD) between two fiber bundle volumes, and the current distance between two fiber bundle volumes. We also discuss an interactive fiber track comparison visualization toolkit we have developed based on the three proposed fiber difference metrics and have tested on six widely-used fiber tracking algorithms. To show the effectiveness and robustness of our metrics and visualization toolkit, we present results on both synthetic data and high resolution monkey brain DT-MRI data. Our toolkit can be used for testing the noise effects on fiber tracking analysis and visualization and to quantify the difference between any pair of DT-MRI techniques, compare single subjects within an image atlas.

A precise knowledge of the microstructures of the myocardium such as myocyte organization and myofiber orientation is necessary to better understand material and functional properties of the tissue. By characterizing the diffusion of water exerted by its molecular environment, magnetic resonance diffusion tensor imaging has emerged as a viable alternative to conventional histology for mapping tissue fibers and offers advantages of being nondestructive, relatively convenient, and inherently 3D. This chapter presents assessments and modeling of myocardial structures via diffusion tensor imaging, including their principles, validation, applications, and potential directions for future development.

Background Previous studies in aging animals have shown that amyloid-beta protein (Aβ) accumulates and its transporters, low-density lipoprotein receptor-related protein-1 (LRP-1) and the receptor for advanced glycation end products (RAGE) are impaired during hydrocephalus. Furthermore, correlations between astrocytes and Aβ have been found in human cases of normal pressure hydrocephalus (NPH) and Alzheimer's disease (AD). Because hydrocephalus occurs frequently in children, we evaluated the expression of Aβ and its transporters and reactive astrocytosis in animals with neonatal hydrocephalus. Methods Hydrocephalus was induced in neonatal rats by intracisternal kaolin injections on post-natal day one, and severe ventriculomegaly developed over a three week period. MRI was performed on post-kaolin days 10 and 21 to document ventriculomegaly. Animals were sacrificed on post-kaolin day 21. For an age-related comparison, tissue was used from previous studies when hydrocephalus was induced in a group of adult animals at either 6 months or 12 months of age. Tissue was processed for immunohistochemistry to visualize LRP-1, RAGE, Aβ, and glial fibrillary acidic protein (GFAP) and with quantitative real time reverse transcriptase polymerase chain reaction (qRT-PCR) to quantify expression of LRP-1, RAGE, and GFAP. Results When 21-day post-kaolin neonatal hydrocephalic animals were compared to adult (6–12 month old) hydrocephalic animals, immunohistochemistry demonstrated levels of Aβ, RAGE, and LRP-1 that were substantially lower in the younger animals; in contrast, GFAP levels were elevated in both young and old hydrocephalic animals. When the neonatal hydrocephalic animals were compared to age-matched controls, qRT-PCR demonstrated no significant changes in Aβ, LRP-1 and RAGE. However, immunohistochemistry showed very small increases or decreases in individual proteins. Furthermore, qRT-PCR indicated statistically significant increases in GFAP. Conclusion Neonatal rats with and without hydrocephalus had low expression of Aβ and its transporters when compared to adult rats with hydrocephalus. No statistical differences were observed in Aβ and its transporters between the control and hydrocephalic neonatal animals.

Background The neurobiology of panic disorder is incompletely understood. The aim of this study was to determine if functional abnormalities of the putamen occur in panic disorder. Methods Activation patterns of 12 female subjects with panic disorder were compared to 18 female healthy controls using functional MRI at 3 T. A motor activation paradigm was used to probe putamen function. Results A complex motor activation paradigm for the non-dominant hand revealed decreased activation of the bilateral putamen among subjects with panic disorder. Limitations The sample size was a relatively small cohort of non-depressed females. Further, some panic disorder subjects were taking medications and/or had comorbid conditions. However, second-level regression analyses did not reveal any correlations between medication use or comorbidity and activation patterns demonstrated by the non-dominant hand complex task. Finally, we used a post-hoc approach to determine the magnitude of global fMRI signal as a surrogate index of the global cerebral blood flow as a means of controlling for possible confounds from reduction of BOLD signal secondary to cerebral vasoconstriction resulting from possible hyperventilation among panic subjects. A more compelling approach would have been to record the respiratory data from subjects during scanning. Conclusions Our findings suggest that putamen dysfunction occurs in at least some cases of panic disorder. We also provide preliminary evidence that a complex motor task for the non-dominant hand is a useful probe of putamen function in this disorder.

The objective of this study was to evaluate an injectable, in situ crosslinkable elastin-like polypeptide (ELP) gel for application to cartilage matrix repair in critically sized defects in goat knees. One cylindrical, osteochondral defect in each of seven animals was filled with an aqueous solution of ELP and a biocompatible, chemical crosslinker, while the contralateral defect remained unfilled and served as an internal control. Joints were sacrificed at 3 (n = 3) or 6 (n = 4) months for MRI, histological, and gross evaluation of features of biomaterial performance, including integration, cellular infiltration, surrounding matrix quality, and new matrix in the defect. At 3 months, ELP-filled defects scored significantly higher for integration by histological and gross grading compared to unfilled defects. ELP did not impede cell infiltration but appeared to be partly degraded. At 6 months, new matrix in unfilled defects outpaced that in ELP-filled defects and scored significantly better for MRI evidence of adverse changes, as well as integration and proteoglycan-containing matrix via gross and histological grading. The ELP-crosslinker solution was easily delivered and formed stable, well-integrated gels that supported cell infiltration and matrix synthesis; however, rapid degradation suggests that ELP formulation modifications should be optimized for longer-term benefits in cartilage repair applications.

Models of corticostriatal motor circuitry have focused on the role of the circuit in the hemisphere of the motor cortex providing primary control (contralateral to the movement). We used functional magnetic resonance imaging and functional connectivity analyses to study circuit function in both the controlling and noncontrolling hemispheres. During the completion of a unilateral motor task with either hand, each putamen nucleus demonstrated strong coactivation with structures in both hemispheres. The putamen in the noncontrolling hemisphere (ipsilateral to the movement) coactivated more strongly with the controlling motor cortex than with the noncontrolling cortex. These findings suggest that the two corticostriatal circuits are functionally integrated. New circuit models based on functional connectivity may need to be developed.

The orientation of MRI-measured diffusion tensor in the myocardium has been directly correlated to the tissue fiber direction and widely characterized. However, the scalar anisotropy indexes have mostly been assumed to be uniform throughout the myocardial wall. The present study examines the fractional anisotropy (FA) as a function of transmural depth and circumferential and longitudinal locations in the normal sheep cardiac left ventricle. Results indicate that FA remains relatively constant from the epicardium to the midwall and then decreases (25.7%) steadily toward the endocardium. The decrease of FA corresponds to 7.9% and 12.9% increases in the secondary and tertiary diffusion tensor diffusivities, respectively. The transmural location of the FA transition coincides with the location where myocardial fibers run exactly circumferentially. There is also a significant difference in the midwall-endocardium FA slope between the septum and the posterior or lateral left ventricular free wall. These findings are consistent with the cellular microstructure from histological studies of the myocardium and suggest a role for MR diffusion tensor imaging in characterization of not only fiber orientation but, also, other tissue parameters, such as the extracellular volume fraction.

Tagged MRI and finite-element (FE) analysis are valuable tools in analyzing cardiac mechanics. To determine systolic material parameters in three-dimensional stress-strain relationships, we used tagged MRI to validate FE models of left ventricular (LV) aneurysm. Five sheep underwent anteroapical myocardial infarction (25% of LV mass) and 22 wk later underwent tagged MRI. Asymmetric FE models of the LV were formed to in vivo geometry from MRI and included aneurysm material properties measured with biaxial stretching, LV pressure measurements, and myofiber helix angles measured with diffusion tensor MRI. Systolic material parameters were determined that enabled FE models to reproduce midwall, systolic myocardial strains from tagged MRI (630 ± 187 strain comparisons/animal). When contractile stress equal to 40% of the myofiber stress was added transverse to the muscle fiber, myocardial strain agreement improved by 27% between FE model predictions and experimental measurements (RMS error decreased from 0.074 ± 0.016 to 0.054 ± 0.011, P < 0.05). In infarct border zone (BZ), end-systolic midwall stress was elevated in both fiber (24.2 ± 2.7 to 29.9 ± 2.4 kPa, P < 0.01) and cross-fiber (5.5 ± 0.7 to 11.7 ± 1.3 kPa, P = 0.02) directions relative to noninfarct regions. Contrary to previous hypotheses but consistent with biaxial stretching experiments, active cross-fiber stress development is an integral part of LV systole; FE analysis with only uniaxial contracting stress is insufficient. Stress calculations from these validated models show 24% increase in fiber stress and 115% increase in cross-fiber stress at the BZ relative to remote regions, which may contribute to LV remodeling.

Mechanical factors such as deformation and strain are thought to play important roles in the maintenance, repair, and degeneration of soft tissues. Determination of soft tissue static deformation has traditionally only been possible at a tissue's surface, utilizing external markers or instrumentation. Texture correlation is a displacement field measurement technique which relies on unique image patterns within a pair of digital images to track displacement. The technique has recently been applied to MR images, indicating the possibility of high-resolution displacement and strain field determination within the mid-substance of soft tissues. However, the utility of MR texture correlation analysis may vary amongst tissue types depending on their underlying structure, composition, and contrast mechanism, which give rise to variations in texture with MRI. In this study, we investigate the utility of a texture correlation algorithm with first-order displacement mapping terms for use with MR images, and suggest a novel index of image "roughness" as a way to decrease errors associated with the use of texture correlation for intra-tissue strain measurement with MRI. We find that a first-order algorithm can significantly reduce strain measurement error, and that an image "roughness" index correlates with displacement measurement error for a variety of imaging conditions and tissue types.

Computer models have been used to study cardiac conduction since the late 1970s. 1 At that time, computational power limited investigations to simple geometries corresponding to single fibers or monolayer sheets of cells. With the evolution of computer technologies, computational models of the heart have become three-dimensional and increasingly more realistic, 2 3 4 allowing a wider range of investigations to elucidate fundamental relations of a set of biophysic parameters and the underlying potential distributions and current flow.

Previous NMR microimaging studies at 360 MHz have demonstrated a clear differentiation between the nucleus and cytoplasm in isolated single neurons. In particular, theT 2 of the cell nucleus is ∼2.5 times larger than that of the cytoplasm. In order to determine the magnitude of possibleT 2 * influences on these observations, images of single cells have been obtained at 500 MHz using spin-echo and line-narrowing sequences. Comparison of the images acquired by the two sequences, and of the spin-echo images at 360 and 500 MHz, imply that anyT 2 * contributions are relatively small. Consequently, the measuredT 2 differences in spin-echo imaging represent a true difference in theT 2 relaxation in the two cellular compartments.

This article investigates the quantitative predictive capabilities of region-specific models by comparing experimental electrograms obtained from in vivo mapping of the ventricular free wall with those obtained through simulation of a region specific three-dimensional bidomain model that incorporates measured fiber orientations. Epicardial electrograms were recorded from canine left ventricles during and after unipolar pacing using a 528-channel electrode plaque. Fiber directions throughout the tissue were estimated from diffusion-weighted MRI and from pace mapping. Electrograms were computed in the bidomain model with experimentally derived properties during paced activations at the same spatiotemporal resolution as those recorded experimentally. Epicardial potentials from model and experiment were directly compared, and sensitivities of these comparisons to reference electrode location and to the choice of material properties were analyzed. The comparisons performed here demonstrate, that (1) the stimulus artifact can be used to estimate the in vivo myocardial fiber architecture, (2) the correlation between simulated and experimental electrograms decreases with increasing pacing depth, and (3) the quantitative comparisons between bidomain model and experimental data are sensitive to both the description of the fiber architecture, and the location of the unipolar reference electrode, but relatively insensitive to moderate changes in the bidomain conductivities. © 2002 Biomedical Engineering Society.

Nonmonoexponential MR diffusion decay behavior has been observed at high diffusion-weighting strengths for cell aggregates and tissues, including the myocardium; however, implications for myocardial MR diffusion tenser imaging are largely unknown. In this study, a slow-exchange-limit, two- component diffusion tenser model was fitted to diffusion- weighted images obtained in isolated, perfused rat hearts. Results indicate that there are at least two distinct components of anisotropic diffusion, characterized by a "fast" component whose principal diffusivity is comparable to that of the perfusate, and a highly anisotropic "slow" component. It is speculated that the two components correspond to tissue compartments and have a general agreement with the orientations of anisotropy, or fiber orientations, in the myocardium. Moreover, consideration of previous studies of myocardial diffusion suggests that the presently observed fast component may likely be dominated by diffusion in the vascular space, whereas the slow component may include the intracellular and interstitial compartments. The implications of the results for myocardial fiber orientation mapping and limitations of the current two-component model used are also discussed. (C) 2001 Wiley-Liss, Inc

Damage to the meniscus after trauma or injury is associated with detrimental changes in joint function that can lead to pain, disability, and degenerative joint changes. Recently, tissue engineering strategies for meniscal repair have been suggested including using biocompatible grafts as a substrate for regeneration, and cellular supplementation to promote remodeling and healing. Little is known, however, about the contributions of these novel repair strategies to restoration of normal meniscal function. Biomechanical factors play a role in the design and synthesis of tissue engineered biomaterials and bioreactors, and also are important for evaluating the efficacy of these new strategies for restoring normal meniscal function. In this report, an overview is presented of biomechanical factors that are critical to meniscal function followed by a review of biomechanical considerations for the design and evaluation of tissue engineered strategies for meniscal repair. Recommendations for future study of biomechanical factors in tissue engineered meniscal repair also are provided.

Functional mapping of the rat somatosensory cortex was performed with T2*-sensitized MRI using a forepaw electrical stimulation model in alpha-chloralose-anesthetized rats at 7 T under both normocapnia and mild hyperventilation-induced hypocapnia. A highly localized activation area, consistent with the known somatosensory cortical region, was detected in all seven animals studied during hypocapnia and in five of the same animals during normocapnia. Quantitatively, hypocapnia was found to significantly increase both the size of the fMRI activation area (3.4 +/- 0.6 mm2 versus 1.5 +/- 0.6 mm2 in normocapnia, mean +/- standard error, n = 7, P0.03) and the average fMRI signal intensity increase (3.4 +/- 0.6% versus 2.7 +/- 0.4%, n = 5, P0.05). The increased sensitivity of fMRI to functional activation may reflect a widened arterial-venous oxygenation difference resulting from an increased effective oxygen extraction during hyperventilation. The dependence of the fMRI response on the ventilation state underscores the need to control for physiological parameters in animal fMRI studies.

The apparent diffusion coefficient (ADC) of water was measured in single Aplysia californica neurons by using NMR microscopy encoded in each of two perpendicular gradient directions. Comparisons of the mean ADCs of the gross nuclear and cytoplasmic compartments in five cells, and 50 subregions within these cells, showed no significant difference between the diffusion measurements in the majority of cases. Since anisotropic diffusion would make the ADC dependent on the encoding direction, the results indicate that the ADC in these single neurons is isotropic at the spatial and temporal resolutions used in these studies. Consequently, a single scalar ADC measurement is sufficient for characterizing the ADC in these cells, hence reducing the acquisition time and measurement complexity that would have been required had the ADC been anisotropic.

Nuclear magnetic resonance (NMR) characteristics of water in perfused single neurons undergoing a 20% hypotonic perturbation were examined quantitatively using NMR microscopy. The transverse relaxation times (T2) in the cytoplasm and nucleus increased by 24.0 +/- 8.5% (average +/- SE, n = 8) and 29.7 +/- 5.3% (n = 6), respectively, whereas the apparent diffusion coefficients (ADC) showed no significant change. These findings are consistent with the behaviors of a perfect osmometer and with accepted molecular relaxation and diffusion models and have significant impacts on current views of properties of cellular water. Furthermore, the results suggest that the increase of tissue intracellular-to-extracellular volume ratio during cell swelling is the predominant mechanism underlying the ADC reduction in acute brain ischemia. These data are the first direct quantitative measurements of the NMR characteristics of water in the cytoplasm and nucleus of single cells undergoing physiological perturbations and may lead to an improved diagnostic capability for NMR imaging in a variety of disease states.

We have used an isolated neuronal preparation of single Aplysia californica L7 cells to study the effects of development on the nuclear magnetic resonance (NMR) characteristics, T2 spin-spin relaxation rate (RT2) and apparent diffusion coefficient (D), of intracellular water within the nuclear and cytoplasmic compartments. These studies demonstrate a significant correlation of animal weight, but not age, with RT2. On the other hand, D was not significantly different as a function of either age or weight in single L7 neurons. Demonstration of maturation dependence in single cells is important in understanding the cellular origins of developmental effects on NMR characteristics in cell assemblies such as brain.

NMR measurements of anisotropic diffusion were studied using a three-dimensional random-walk model. It was found that the apparent diffusion coefficient can be expressed in a canonical form as the product of a diagonal matrix, an orthonormal rotation matrix, and a vector representing the encoding magnetic field gradient. The diffusion coefficient can be interpreted as the sum of the corresponding coefficients measured along the principal diffusion axes, weighted by the squares of the directional cosines of the encoding direction with respect to the principal axes. The analysis revealed that determining the orientation of anisotropy, in a cylindrically symmetric system, requires a minimum of four diffusion measurements. A special pulse sequence which minimized gradient cross-terms and possible restricted diffusion effects was used to characterize diffusion anisotropy in cut chicken gizzards. Diffusion coefficients parallel to the muscle fibers were found to be approximately two to three times larger than those in the transverse direction. Furthermore, the method was successful in detecting the angular change when the sample was rotated by 30 degrees. Results indicate that the proposed approach to measure fiber orientation is valid and may be used to improve the time efficiency of diffusion anisotropy measurements.

O. M. Abdullah, S. G. Drakos, D. R. Verma, J. Stehlik, A. G. Kfoury, C. H. Selzman, C. Myrick, G. Russel, D. Y. Li, and E. W. Hsu Bioengineering, University of Utah, Salt Lake City, UT, United States, Small Animal Core Research, University of Utah, Salt Lake City, UT, United States, UTAH Cardiac Transplant Program, University of Utah & Intermountain Medical Center, Salt Lake City, UT, United States, Intermountain Donor Services, Salt Lake City, UT, United States

ObjectiveLinear repair of left ventricular aneurysm has been performed with mixed clinical results. By using finite element analysis, this study evaluated the effect of this procedure on end-systolic stress.MethodsNine sheep underwent myocardial infarction and aneurysm repair with a linear repair (13.4 ± 2.3 weeks postmyocardial infarction). Satisfactory magnetic resonance imaging examinations were obtained in 6 sheep (6.6 ± 0.5 weeks postrepair). Finite element models were constructed from in vivo magnetic resonance imaging-based cardiac geometry and postmortem measurement of myofiber helix angles using diffusion tensor magnetic resonance imaging. Material properties were iteratively determined by comparing the finite element model output with systolic tagged magnetic resonance imaging strain measurements.ResultsAt the mid-wall, fiber stress in the border zone decreased by 39% (sham = 32.5 ± 2.5 kPa, repair = 19.7 ± 3.6 kPa, P = .001) to the level of remote regions after repair. In the septum, however, border zone fiber stress remained high (sham = 31.3 ± 5.4 kPa, repair = 23.8 ± 5.8 kPa, P = .29). Cross-fiber stress at the mid-wall decreased by 41% (sham = 13.0 ± 1.5 kPa, repair = 7.7 ± 2.1 kPa, P = .01), but cross-fiber stress in the un-excluded septal infarct was 75% higher in the border zone than remote regions (remote = 5.9 ± 1.9 kPa, border zone = 10.3 ± 3.6 kPa, P < .01). However, end-diastolic fiber and cross-fiber stress were not reduced in the remote myocardium after plication.ConclusionWith the exception of the retained septal infarct, end-systolic stress is reduced in all areas of the left ventricle after infarct plication. Consequently, we expect the primary positive effect of infarct plication to be in the infarct border zone. However, the amount of stress reduction necessary to halt or reverse nonischemic infarct extension in the infarct border zone and eccentric hypertrophy in the remote myocardium is unknown.

Purpose of study: To develop an experimental model of degenerative disc disease without direct intervention of the intervertebral disc (IVD). Our hypothesis was that inducing vertebral body damage at sites adjacent to the IVD would modify nutrient transport through the end plates and thus initiate degeneration in a manner relevant to degenerative disc disease in humans. Methods used: Vertebral body defects were created in lumbar motion segments of minipigs (10 experimental and 7 control levels, n=3). A high-speed drill with a diamond burr (3 to 5 mm diameter) was used to remove bone adjacent to the inferior and superior end plates of the experimental IVDs. A resorbable calcium phosphate bone cement was then packed into the defects. Spines were harvested at 3, 6 and 9 months and imaged in the sagittal plane with high-resolution magnetic resonance imaging (MRI; 7.1 T). Four blinded observers graded MRI appearance with the scheme of Thompson and coworkers (1990; 1 = nondegenerate, 5 = severely degenerate). In addition, MRI data sets were analyzed to determine changes in disc height and signal intensity for regions corresponding to the nucleus pulposus (NP) and anterior and posterior anulus fibrosus (AF). After imaging, individual motion segments were processed for routine histology (toluidine blue, hemotoxylin and eosin) and immunohistochemistry to detect matrix proteins including type II collagen and decorin. of findings: Higher MRI grades of degeneration were observed in all experimental IVD regions at 9 months after surgery. Reviewer grades were significantly higher for both NP (average of four graders = 1.8 vs. 3.8, control vs. experimental) and AF at 9 months after surgery (1.7 vs. 3.4). There was some evidence of end plate changes beginning at 3 months after surgery (1.5 vs. 2.3, control vs. experimental), which became quite dramatic at 9 months (1.4 vs. 4.5). No differences were noted in disc height between control and experimental IVDs at any region. However, decreases in MRI signal intensity (normalized to maximum values) were noted in experimental IVDs, particularly in the anterior AF (mean, 39%) and NP at 9 months (15%). Decreased signal intensity is a hallmark of IVD degeneration in the human, suggesting loss of hydration. Upon histological examination, there was evidence of inward bulging of the inner AF, end plate damage and decreased NP staining. Immunostaining demonstrated increased collagen type II in the experimental sections compared with controls, although few consistent changes were noted in the expression of decorin. Increases in the levels of both collagen type II and decorin have been demonstrated in other animal models of IVD degeneration and in human disease. Relationship between findings and existing knowledge: Overall, the changes observed in this experimental model confirm some key features of human IVD degeneration, including increased radiographic grade of degeneration, decreased MRI intensity and morphological and histological changes, such as increased staining for collagen type II and decreased staining of the NP. Holm and coworkers (1999) demonstrated that a controlled vertebral end plate defect will induce compositional and morphological evidence of disc degeneration in an animal model. Findings of the current study demonstrate that disruption of the vertebral body alone, without controlled violation of the end plate, may serve to initiate disc degeneration without direct, surgical injury of the IVD. Overall significance of findings: The mechanism for the onset of degeneration is not known but may relate to a role of vertebral body damage in altering nutrient transport across the end plate. Disclosures: No disclosures. Conflict of interest: William J. Richardson, grant research support, DePuy Orthobiologics.

The apparent diffusion coefficient (ADC) of water after regional myocardial ischemia was measured in isolated, perfused rabbit hearts by using magnetic resonance imaging (MRI) techniques. After ligation of the left anterior descending coronary artery, the ADC of the nonperfused region showed a gradual but significant decreasing trend over time, whereas that of the normally perfused myocardium remained constant. Morphological analysis revealed that the ADC decrease reflected the expansion of a subregion of reduced ADC within the nonperfused myocardium. The dynamics of the diffusion change and the morphological progression of the affected tissue suggest that the ADC decrease may be linked to the onset of myocardial infarction, which is known to involve myocyte swelling. The ADC reduction provides a potentially valuable MRI tissue-contrast mechanism for noninvasively determining the viability of the ischemic myocardium and assessing the dynamics of acute myocardial infarction.