An Imaging Protocol to Discriminate Specialized Conduction Tissue During Congenital Heart Surgery.

We performed preclinical validation of intraoperative fiber-optic confocal microscopy (FCM) and assessed its safety and efficacy in an ovine model of the pediatric heart. Intraoperative imaging was performed using an FCM system (Cellvizio, Mauna Kea Technology, Paris, France) with specialized imaging miniprobe (GastroFlex UHD, Mauna Kea Technologies). Before imaging, we applied an extracellular fluorophore, sodium fluorescein, to fluorescently label extracellular space. We imaged arrested hearts of ovine (1-6 months) under cardiopulmonary bypass. Image sequences (1-10 seconds duration) were acquired from regions of the sinoatrial and atrioventricular node, as well as subepicardial and subendocardial working myocardium from atria and ventricle. The surgical process was evaluated for integration of the imaging protocol during the operative procedure. In addition, fluorescein cardiotoxicity studies (n = 3 animals) were conducted by comparing electrocardiogram (PR and QRS intervals) and ejection fraction at baseline and after topical application of fluorescein at 1:10, 1:100, and 1:1000 dilutions on a beating ovine heart. Our studies suggest that intraoperative FCM can be used to identify regions associated with specialized conducting tissue in ovine hearts in situ. The imaging protocol was integrated with conventional open heart surgical procedures with minimal changes to the operative process. Application of fluorescein in varying concentrations did not affect the normalized PR interval, QRS interval, and ejection fraction. These preclinical validation studies demonstrated both safety and efficacy of the proposed intraoperative imaging approach. The studies constitute an important step toward first-in-human clinical trials.

Evidence for widespread structural brain changes in glaucoma: a preliminary voxel-based MRI study.

PURPOSE: To investigate structural brain changes in patients with glaucoma. METHODS: High-resolution T1-weighted anatomical brain magnetic resonance images (MRI) were collected in 15 patients with glaucoma of varying severity and in 15 age-, race-, and sex-matched controls. Exclusion criteria included neurological disease, another disorder which could affect the visual field, and a score of less than 25 on the mini-mental status examination. The scans were analyzed with an automatic volumetric MRI technique to measure the volumes of 93 structures in each brain. Analyses of covariance with age as a covariate were carried out to identify structures that differed significantly between the two groups (i.e., glaucoma versus normal control). The volumes of all brain structures in the group of 15 glaucoma patients were also correlated with clinical measures of disease severity. Linear multivariate regression analyses were conducted to determine the significance of these relationships. RESULTS: Five structures differed significantly between the two groups (P < 0.05). These structures included the right and left inferior occipital gyri and the right middle occipital gyrus, right inferior temporal gyrus, and right occipital lobe white matter. Interestingly, all of these structures were larger in the glaucoma group than in the control group. Within the group of glaucoma patients, 38% of all brain structures had independent associations between decreasing volume and more severe disease in multivariate regression analysis. CONCLUSIONS: These results suggest that patients with glaucoma undergo widespread and complex changes in cortical brain structure and that the extent of these changes correlates with disease severity.

Sex chromosomes and the brain: a study of neuroanatomy in XYY syndrome.

AIM: To assess global and regional brain matter variations associated with XYY syndrome by comparison with Klinefelter syndrome and typical development. METHODS: We used two conceptually distinct voxel-based magnetic resonance imaging methods to examine brain structure in young males with XYY syndrome: (1) volumetric comparison to assess global grey and white matter volumes and (2) support vector machine-based multivariate pattern classification analysis to assess regional neuroanatomy. We assessed verbal, non-verbal, and spatial abilities with the Differential Ability Scales (DAS), and we measured autism diagnostic criteria in eight males with XYY syndrome using the Social Responsiveness Scale and the Autism Diagnostic Interview-Revised (ADI-R). RESULTS: A comparison of 36 typically developing males (mean age 11 y, SD 1 y 9 mo), 31 males with Klinefelter syndrome (mean age 9 y 8 mo, SD 1 y 8 mo), and eight males with XYY syndrome (mean age 11 y 6 mo, SD 1 y 11 mo) showed that total white and grey matter volumes were significantly, or nearly significantly, higher in males with XYY syndrome than in males belonging to the other two groups (grey matter: XYY males vs typically developing males, p<0.006; XYY vs males with Klinefelter syndrome, p<0.001; white matter: XYY males vs typically developing males, p=0.061; XYY males vs males with Klinefelter syndrome, p=0.004). Voxel-based multivariate pattern classification analysis indicates that, after controlling for global volumes, regional brain variations in XYY syndrome are more like those found in Klinefelter syndrome than those occurring in typical development. Further, visualization of classification parameters suggests that insular and frontotemporal grey matter and white matter, including known language areas, are reduced in males with XYY syndrome, similar to what is seen in Klinefelter syndrome. In males with XYY syndrome, DAS verbal and non-verbal scores were significantly lower than in typically developing participants (both p<0.001). DAS scores were not significantly different between XYY and Klinefelter syndrome groups. In five of eight males with XYY syndrome, the Social Responsiveness Scale score exceeded the cut-off for a likely diagnosis of autism spectrum disorder (ASD). In three of eight males with XYY syndrome, the ADI-R score met the cut-off for ASD diagnosis; in another two, ADI-R scores within the social and communication domains met the cut-off values for a diagnosis of ASD. INTERPRETATION: The results suggest that genetic variations associated with XYY syndrome result in increased brain matter volumes, a finding putatively related to the increased frequency of ASDs in individuals with this condition. In addition, frontotemporal grey and white matter reductions in XYY syndrome provide a likely neuroanatomical correlate for observed language impairments.

Neuroanatomical phenotype of Klinefelter syndrome in childhood: a voxel-based morphometry study.

Klinefelter syndrome (KS) is a genetic disorder characterized by a supernumerary X chromosome. As such, KS offers a naturally occurring human model for the study of both X-chromosome gene expression and androgen on brain development. Previous neuroimaging studies have revealed neuroanatomical variations associated with KS, but have differed widely with respect to subject inclusion criteria, including mosaicism, pubertal status, and history of testosterone replacement therapy (TRT), all factors likely to influence neurodevelopment. We conducted a voxel-based morphometry study of regional gray and white matter (GM and WM, respectively) volumes in 31 KS males (mean age, 9.69 ± 1.70 years) and 36 typically developing (TD) male controls (10.99 ± 1.72 years). None of the participants with KS had received TRT, and all were prepubertal and had nonmosaic 47,XXY karyotypes. After controlling for age, males with KS showed trends (0.05 < p < 0.10) for significantly reduced total gray matter volume (TGMV) and total white matter volume (TWMV), relative to TD males. After controlling for TGMV and age, the KS group had significantly increased sensorimotor and parietal-occipital GM and significantly reduced amygdalar, hippocampal, insular, temporal, and inferior frontal GM relative to TD controls. After controlling for TWMV and age, the KS group had significantly increased left parietal WM as well as significantly reduced frontal and temporal WM. These findings are indicative of a characteristic prepubertal neuroanatomical phenotype that may be associated with cognitive-behavioral features of KS. This work offers new insight into the relationships among X-chromosome gene expression, neuroanatomy, and cognitive-behavioral functions impaired in KS, including language and attention.

Atypical functional brain activation during a multiple object tracking task in girls with Turner syndrome: neurocorrelates of reduced spatiotemporal resolution.

Turner syndrome is associated with spatial and numerical cognitive impairments. We hypothesized that these nonverbal cognitive impairments result from limits in spatial and temporal processing, particularly as it affects attention. To examine spatiotemporal attention in girls with Turner syndrome versus typically developing controls, we used a multiple object tracking task during functional magnetic resonance (fMRI) imaging. Participants actively tracked a target among six distracters or passively viewed the animations. Neural activation in girls with Turner syndrome during object tracking overlapped with but was dissimilar to the canonical frontoparietal network evident in typically developing controls and included greater limbic activity. Task performance and atypical functional activation indicate anomalous development of cortical and subcortical temporal and spatial processing circuits in girls with Turner syndrome.

Toward a practical protocol for human optic nerve DTI with EPI geometric distortion correction.

PURPOSE: To develop a practical protocol for diffusion tensor imaging (DTI) of the human optic nerve with echo planar imaging (EPI) geometric distortion correction. MATERIALS AND METHODS: A conventional DTI protocol was modified to acquire images with fat and cerebrospinal fluid (CSF) suppression and field inhomogeneity maps of contiguous coronal slices covering the whole brain. The technique was applied to healthy volunteers and multiple sclerosis patients with and without a history of unilateral optic neuritis. DTI measures and optic nerve tractography before and after geometric distortion correction were compared. Diffusion measures from left and right or from affected and unaffected eyes in different subject cohorts were reported. RESULTS: The image geometry after correction closely resembled reference anatomical images. Optic nerve tractography became feasible after distortion correction. The diffusion measures from the healthy volunteers were in good agreement with the literature. Statistically significant differences were found in the fractional anisotropy and orthogonal eigenvalues between affected and unaffected eyes in optic neuritis patients with poor recovery. The diffusion measures before and after geometric distortion correction were not significantly different. For cohorts without optic neuritis, the difference between diffusion measures from left and right eyes was not statistically significant. CONCLUSION: The proposed technique could provide a practical DTI protocol to study the human optic nerve.

Estimation of mutual information objective function based on Fourier shift theorem: an application to eddy current distortion correction in diffusion tensor imaging.

Diffusion tensor imaging requires correction of eddy current distortion in diffusion-weighted images. An effective retrospective correction approach is to transform a diffusion-weighted image to maximize the mutual information (MI) between the transformed diffusion-weighted image and the corresponding T2-weighted image. In the literature, either linear interpolation or partial volume interpolation is applied to estimate the MI objective function. However, these interpolation methods induce artifacts to the MI objective function, thus compromising correction results. In this work, the MI objective function is estimated based on interpolation using Fourier shift theorem. This method eliminates the artifacts incurred with the aforementioned interpolation methods. The algorithm is further improved by approximating pixel values using their nearest neighbors in the up-sampled spatial domain, resulting in dramatically increased computational efficiency without compromising the correction results. The effects of varying the number of quantization levels and using Parzen window filtering to smooth the MI objective function are also investigated to obtain optimized algorithm parameters. The diffusion tensor image quality after applying the proposed distortion correction method is significantly improved visually.

Phase labeling using sensitivity encoding (PLUS): data acquisition and image reconstruction for geometric distortion correction in EPI.

Geometric distortion caused by magnetic field inhomogeneity is generally an inevitable tradeoff for fast MRI acquisitions using echo-planar imaging. Most of the existing distortion-correction techniques require separate scans for field maps in order to correct the distortion contained in a measurement. A drawback of these current techniques is that the field map scans and the measurement can capture different patient positions, which invalidates the stationary condition. A new method was developed in this work to correct geometric distortion by using local phase shifts derived directly from the measurement itself, without the need of extra field map scans. This self-sufficient method takes advantage of parallel imaging and k-space trajectory modification to produce multiple images from a single acquisition. The measurement is also used to derive sensitivity maps for parallel imaging reconstruction. The derived phase shifts are retrospectively applied to the measurement for correction of geometric distortion in the measurement itself. The proposed method was successfully demonstrated using experimental data from a phantom and a human brain.

Revisiting anaplastic astrocytomas II: further characterization of an expansive growth pattern with visually enhanced diffusion tensor imaging.

PURPOSE: To seek to distinguish and visualize the different magnetic resonance imaging (MRI) growth patterns among malignant gliomas utilizing visually enhanced diffusion tensor imaging (DTI). MATERIALS AND METHODS: Nineteen consecutive patients undergoing image-guided resection of a newly diagnosed malignant glioma underwent add-on acquisition of DTI data based on an Institutional Review Board (IRB)-approved imaging protocol during preoperative MRI scans for routine intraoperative image guidance. Tumor growth patterns were assigned to expansive or mixed/infiltrative classes as described in the companion article (24). Infiltrating tumors were WHO Grade IV astrocytomas and all expansive tumors were either WHO Grade III astrocytomas or WHO Grade II astrocytomas. DTI-based white matter tractography was conducted and the DTI data were fused with anatomical images using an in-house software package we developed to enhance the visualization of the tumor/fiber interface. In one case additional analysis was performed with 2D multivoxel (1)H-MRSI utilizing a 2D chemical shift imaging (CSI) technique to corroborate the nature of this interface. RESULTS: Out of the 19 tumor patients studied, 11 had infiltrative tumors and the other 8 had expansive tumors. While less clear with 2D axial diffusion color maps, visually enhanced 3D reconstructions of the tumor/fiber interface successfully corroborated distinctive growth patterns. This was particularly evident when viewed in 3D video loops of each tumor/fiber interface. CONCLUSION: We have successfully developed software that visually enhances the anatomic details of the tumor/fiber interface in patients with anaplastic astrocytomas. These data support the existence of a subgroup of patients within the WHO Grade III classification with expansive tumors and a significantly better prognosis.

Design of (Gd-DO3A)n-polydiamidopropanoyl-peptide nucleic acid-D(Cys-Ser-Lys-Cys) magnetic resonance contrast agents.

We hypothesized that chelating Gd(III) to 1,4,7-tris(carboxymethylaza)cyclododecane-10-azaacetylamide (DO3A) on peptide nucleic acid (PNA) hybridization probes would provide a magnetic resonance genetic imaging agent capable of hybridization to a specific mRNA. Because of the low sensitivity of Gd(III) as an magnetic resonance imaging (MRI) contrast agent, a single Gd-DO3A complex per PNA hybridization agent could not provide enough contrast for detection of cancer gene mRNAs, even at thousands of mRNA copies per cell. To increase the Gd(III) shift intensity of MRI genetic imaging agents, we extended a novel DO3An-polydiamidopropanoyl (PDAPm) dendrimer, up to n = 16, from the N-terminus of KRAS PNA hybridization agents by solid phase synthesis. A C-terminal D(Cys-Ser-Lys-Cys) cyclized peptide analog of insulin-like growth factor 1 (IGF1) was included to enable receptor-mediated cellular uptake. Molecular dynamic simulation of the (Gd-DO3A-AEEA)16-PDAP4-AEEA2-KRAS PNA-AEEA-D(Cys-Ser-Lys-Cys) genetic imaging nanoparticles in explicit water yielded a pair correlation function similar to that of PAMAM dendrimers, and a predicted structure in which the PDAP dendron did not sequester the PNA. Thermal melting measurements indicated that the size of the PDAP dendron included in the (DO3A-AEEA)n-PDAPm-AEEA2-KRAS PNA-AEEA-D(Cys-Ser-Lys-Cys) probes (up to 16 Gd(III) cations per PNA) did not depress the melting temperatures (Tm) of the complementary PNA/RNA hybrid duplexes. The Gd(III) dendrimer PNA genetic imaging agents in phantom solutions displayed significantly greater T1 relaxivity per probe (r1 = 30.64 +/- 2.68 mM(-1) s(-1) for n = 2, r1 = 153.84 +/- 11.28 mM(-1) s(-1) for n = 8) than Gd-DTPA (r1 = 10.35 +/- 0.37 mM(-1) s(-1)), but less than that of (Gd-DO3A)32-PAMAM dendrimer (r1 = 771.84 +/- 20.48 mM(-1) s(-1)) (P < 0.05). Higher generations of PDAP dendrimers with 32 or more Gd-DO3A residues attached to PNA-D(Cys-Ser-Lys-Cys) genetic imaging agents might provide greater contrast for more sensitive detection.