Prof Jurgen E Schneider

Research Area: Imaging (macro)
Technology Exchange: In vivo imaging and Magnetic resonance imaging & spectroscopy
Keywords: Magnetic Resonance Imaging (MRI), Magnetic Resonance Spectroscopy (MRS), Ultra-high magnetic fields, Cardiac metabolism, Cardiac structure and Cardiac function
Normal creatine metabolism assessed by cardiac 1H-MRS.

Normal creatine metabolism assessed by cardiac 1H-MRS.

Abnormal creatine metabolism assessed by cardiac 1H-MRS.

Abnormal creatine metabolism assessed by cardiac 1H-MRS.

BMRU

BMRU

  • Development of fast cardiac MRI techniques at ultra-high fields

  • MR Microscopy

  • Development and application of cardiac MRS - see below for examples for (A) normal and (B) abnormal creatine metabolism assessed by cardiac 1H-MRS.

  • Cardiac DTI

Name Department Institution Country
Prof Andrew Blamire Newcastle Magnetic Resonance Centre, Campus for Ageing and Vitality Newcastle University United Kingdom
Prof Mark A. Griswold Department of Radiology - MRI University Hospitals of Cleveland United States
Prof Herbert Köstler Experimental Radiology University of Würzburg Germany
Prof Peter Kohl Imperial College The Heart Science Centre, National Heart and Lung Institute United Kingdom
Prof Nicole Seiberlich Biomedical Engineering Case Western Reserve University United States
Prof Nicolas Smith Imaging Sciences & Biomedical Engineering Division, St Thomas’ Hospital King's College United Kingdom
Prof Alison Noble FREng (MPLS) Oxford University,
Prof Ben Davis Department of Chemistry University of Oxford United Kingdom
Hulley PA, Bishop T, Vernet A, Schneider JE, Edwards JR, Athanasou NA, Knowles HJ. 2017. Hypoxia-inducible factor 1-alpha does not regulate osteoclastogenesis but enhances bone resorption activity via prolyl-4-hydroxylase 2. J Pathol, 242 (3), pp. 322-333. | Show Abstract | Read more

Osteogenic-angiogenic coupling is promoted by the hypoxia-inducible factor 1-alpha (HIF-1α) transcription factor, provoking interest in HIF activation as a therapeutic strategy to improve osteoblast mineralization and treat pathological osteolysis. However, HIF also enhances the bone-resorbing activity of mature osteoclasts. It is therefore essential to determine the full effect(s) of HIF on both the formation and the bone-resorbing function of osteoclasts in order to understand how they might respond to such a strategy. Expression of HIF-1α mRNA and protein increased during osteoclast differentiation from CD14+ monocytic precursors, additionally inducing expression of the HIF-regulated glycolytic enzymes. However, HIF-1α siRNA only moderately affected osteoclast differentiation, accelerating fusion of precursor cells. HIF induction by inhibition of the regulatory prolyl-4-hydroxylase (PHD) enzymes reduced osteoclastogenesis, but was confirmed to enhance bone resorption by mature osteoclasts. Phd2(+/-) murine osteoclasts also exhibited enhanced bone resorption, associated with increased expression of resorption-associated Acp5, in comparison with wild-type cells from littermate controls. Phd3(-/-) bone marrow precursors displayed accelerated early fusion, mirroring results with HIF-1α siRNA. In vivo, Phd2(+/-) and Phd3(-/-) mice exhibited reduced trabecular bone mass, associated with reduced mineralization by Phd2(+/-) osteoblasts. These data indicate that HIF predominantly functions as a regulator of osteoclast-mediated bone resorption, with little effect on osteoclast differentiation. Inhibition of HIF might therefore represent an alternative strategy to treat diseases characterized by pathological levels of osteolysis. © 2017 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.

Teh I, McClymont D, Zdora MC, Whittington HJ, Davidoiu V, Lee J, Lygate CA, Rau C, Zanette I, Schneider JE. 2017. Validation of diffusion tensor MRI measurements of cardiac microstructure with structure tensor synchrotron radiation imaging. J Cardiovasc Magn Reson, 19 (1), pp. 31. | Show Abstract | Read more

BACKGROUND: Diffusion tensor imaging (DTI) is widely used to assess tissue microstructure non-invasively. Cardiac DTI enables inference of cell and sheetlet orientations, which are altered under pathological conditions. However, DTI is affected by many factors, therefore robust validation is critical. Existing histological validation is intrinsically flawed, since it requires further tissue processing leading to sample distortion, is routinely limited in field-of-view and requires reconstruction of three-dimensional volumes from two-dimensional images. In contrast, synchrotron radiation imaging (SRI) data enables imaging of the heart in 3D without further preparation following DTI. The objective of the study was to validate DTI measurements based on structure tensor analysis of SRI data. METHODS: One isolated, fixed rat heart was imaged ex vivo with DTI and X-ray phase contrast SRI, and reconstructed at 100 μm and 3.6 μm isotropic resolution respectively. Structure tensors were determined from the SRI data and registered to the DTI data. RESULTS: Excellent agreement in helix angles (HA) and transverse angles (TA) was observed between the DTI and structure tensor synchrotron radiation imaging (STSRI) data, where HADTI-STSRI = -1.4° ± 23.2° and TADTI-STSRI = -1.4° ± 35.0° (mean ± 1.96 standard deviation across all voxels in the left ventricle). STSRI confirmed that the primary eigenvector of the diffusion tensor corresponds with the cardiomyocyte long-axis across the whole myocardium. CONCLUSIONS: We have used STSRI as a novel and high-resolution gold standard for the validation of DTI, allowing like-with-like comparison of three-dimensional tissue structures in the same intact heart free of distortion. This represents a critical step forward in independently verifying the structural basis and informing the interpretation of cardiac DTI data, thereby supporting the further development and adoption of DTI in structure-based electro-mechanical modelling and routine clinical applications.

Lottonen-Raikaslehto L, Rissanen R, Gurzeler E, Merentie M, Huusko J, Schneider JE, Liimatainen T, Ylä-Herttuala S. 2017. Left ventricular remodeling leads to heart failure in mice with cardiac-specific overexpression of VEGF-B167: echocardiography and magnetic resonance imaging study. Physiol Rep, 5 (6), pp. e13096-e13096. | Show Abstract | Read more

Cardiac-specific overexpression of vascular endothelial growth factor (VEGF)-B167 is known to induce left ventricular hypertrophy due to altered lipid metabolism, in which ceramides accumulate to the heart and cause mitochondrial damage. The aim of this study was to evaluate and compare different imaging methods to find the most sensitive way to diagnose at early stage the progressive left ventricular remodeling leading to heart failure. Echocardiography and cardiovascular magnetic resonance imaging were compared for imaging the hearts of transgenic mice with cardiac-specific overexpression of VEGF-B167 and wild-type mice from 5 to 14 months of age at several time points. Disease progression was verified by molecular biology methods and histology. We showed that left ventricular remodeling is already ongoing at the age of 5 months in transgenic mice leading to heart failure by the age of 14 months. Measurements from echocardiography and cardiovascular magnetic resonance imaging revealed similar changes in cardiac structure and function in the transgenic mice. Changes in histology, gene expressions, and electrocardiography supported the progression of left ventricular hypertrophy. Longitudinal relaxation time in rotating frame (T1ρ ) in cardiovascular magnetic resonance imaging could be suitable for detecting severe fibrosis in the heart. We conclude that cardiac-specific overexpression of VEGF-B167 leads to left ventricular remodeling at early age and is a suitable model to study heart failure development with different imaging methods.

Atzler D, McAndrew DJ, Cordts K, Schneider JE, Zervou S, Schwedhelm E, Neubauer S, Lygate CA. 2017. Dietary Supplementation with Homoarginine Preserves Cardiac Function in a Murine Model of Post-Myocardial Infarction Heart Failure. Circulation, 135 (4), pp. 400-402. | Read more

Al Haj Zen A, Nawrot DA, Howarth A, Caporali A, Ebner D, Vernet A, Schneider JE, Bhattacharya S. 2016. The Retinoid Agonist Tazarotene Promotes Angiogenesis and Wound Healing. Mol Ther, 24 (10), pp. 1745-1759. | Show Abstract | Read more

Therapeutic angiogenesis is a major goal of regenerative medicine, but no clinically approved small molecule exists that enhances new blood vessel formation. Here we show, using a phenotype-driven high-content imaging screen of an annotated chemical library of 1,280 bioactive small molecules, that the retinoid agonist Tazarotene, enhances in vitro angiogenesis, promoting branching morphogenesis, and tubule remodeling. The proangiogenic phenotype is mediated by retinoic acid receptor but not retinoic X receptor activation, and is characterized by secretion of the proangiogenic factors hepatocyte growth factor, vascular endothelial growth factor, plasminogen activator, urokinase and placental growth factor, and reduced secretion of the antiangiogenic factor pentraxin-3 from adjacent fibroblasts. In vivo, Tazarotene enhanced the growth of mature and functional microvessels in Matrigel implants and wound healing models, and increased blood flow. Notably, in ear punch wound healing model, Tazarotene promoted tissue repair characterized by rapid ear punch closure with normal-appearing skin containing new hair follicles, and maturing collagen fibers. Our study suggests that Tazarotene, an FDA-approved small molecule, could be potentially exploited for therapeutic applications in neovascularization and wound healing.

Zak J, Vives V, Szumska D, Vernet A, Schneider JE, Miller P, Slee EA, Joss S, Lacassie Y, Chen E et al. 2016. ASPP2 deficiency causes features of 1q41q42 microdeletion syndrome. Cell Death Differ, 23 (12), pp. 1973-1984. | Show Abstract | Read more

Chromosomal abnormalities are implicated in a substantial number of human developmental syndromes, but for many such disorders little is known about the causative genes. The recently described 1q41q42 microdeletion syndrome is characterized by characteristic dysmorphic features, intellectual disability and brain morphological abnormalities, but the precise genetic basis for these abnormalities remains unknown. Here, our detailed analysis of the genetic abnormalities of 1q41q42 microdeletion cases identified TP53BP2, which encodes apoptosis-stimulating protein of p53 2 (ASPP2), as a candidate gene for brain abnormalities. Consistent with this, Trp53bp2-deficient mice show dilation of lateral ventricles resembling the phenotype of 1q41q42 microdeletion patients. Trp53bp2 deficiency causes 100% neonatal lethality in the C57BL/6 background associated with a high incidence of neural tube defects and a range of developmental abnormalities such as congenital heart defects, coloboma, microphthalmia, urogenital and craniofacial abnormalities. Interestingly, abnormalities show a high degree of overlap with 1q41q42 microdeletion-associated abnormalities. These findings identify TP53BP2 as a strong candidate causative gene for central nervous system (CNS) defects in 1q41q42 microdeletion syndrome, and open new avenues for investigation of the mechanisms underlying CNS abnormalities.

Levelt E, Pavlides M, Banerjee R, Mahmod M, Kelly C, Sellwood J, Ariga R, Thomas S, Francis J, Rodgers C et al. 2016. Ectopic and Visceral Fat Deposition in Lean and Obese Patients With Type 2 Diabetes. J Am Coll Cardiol, 68 (1), pp. 53-63. | Show Abstract | Read more

BACKGROUND: Type 2 diabetes (T2D) and obesity are associated with nonalcoholic fatty liver disease, cardiomyopathy, and cardiovascular mortality. Both show stronger links between ectopic and visceral fat deposition, and an increased cardiometabolic risk compared with subcutaneous fat. OBJECTIVES: This study investigated whether lean patients (Ln) with T2D exhibit increased ectopic and visceral fat deposition and whether these are linked to cardiac and hepatic changes. METHODS: Twenty-seven obese patients (Ob) with T2D, 15 Ln-T2D, and 12 normal-weight control subjects were studied. Subjects underwent cardiac computed tomography, cardiac magnetic resonance imaging (MRI), proton and phosphorus MR spectroscopy, and multiparametric liver MR, including hepatic proton MRS, T1- and T2*-mapping yielding "iron-corrected T1" [cT1]. RESULTS: Diabetes, with or without obesity, was associated with increased myocardial triglyceride content (p = 0.01), increased hepatic triglyceride content (p = 0.04), and impaired myocardial energetics (p = 0.04). Although cardiac structural changes, steatosis, and energetics were similar between the T2D groups, epicardial fat (p = 0.04), hepatic triglyceride (p = 0.01), and insulin resistance (p = 0.03) were higher in Ob-T2D. Epicardial fat, hepatic triglyceride, and insulin resistance correlated negatively with systolic strain and diastolic strain rates, which were only significantly impaired in Ob-T2D (p < 0.001 and p = 0.006, respectively). Fibroinflammatory liver disease (elevated cT1) was only evident in Ob-T2D patients. cT1 correlated with hepatic and epicardial fat (p < 0.001 and p = 0.01, respectively). CONCLUSIONS: Irrespective of body mass index, diabetes is related to significant abnormalities in cardiac structure, energetics, and cardiac and hepatic steatosis. Obese patients with T2D show a greater propensity for ectopic and visceral fat deposition.

Teh I, Burton RA, McClymont D, Capel RA, Aston D, Kohl P, Schneider JE. 2016. Mapping cardiac microstructure of rabbit heart in different mechanical states by high resolution diffusion tensor imaging: A proof-of-principle study. Prog Biophys Mol Biol, 121 (2), pp. 85-96. | Show Abstract | Read more

Myocardial microstructure and its macroscopic materialisation are fundamental to the function of the heart. Despite this importance, characterisation of cellular features at the organ level remains challenging, and a unifying description of the structure of the heart is still outstanding. Here, we optimised diffusion tensor imaging data to acquire high quality data in ex vivo rabbit hearts in slack and contractured states, approximating diastolic and systolic conditions. The data were analysed with a suite of methods that focused on different aspects of the myocardium. In the slack heart, we observed a similar transmural gradient in helix angle of the primary eigenvector of up to 23.6°/mm in the left ventricle and 24.2°/mm in the right ventricle. In the contractured heart, the same transmural gradient remained largely linear, but was offset by up to +49.9° in the left ventricle. In the right ventricle, there was an increase in the transmural gradient to 31.2°/mm and an offset of up to +39.0°. The application of tractography based on each eigenvector enabled visualisation of streamlines that depict cardiomyocyte and sheetlet organisation over large distances. We observed multiple V- and N-shaped sheetlet arrangements throughout the myocardium, and insertion of sheetlets at the intersection of the left and right ventricle. This study integrates several complementary techniques to visualise and quantify the heart's microstructure, projecting parameter representations across different length scales. This represents a step towards a more comprehensive characterisation of myocardial microstructure at the whole organ level.

Redgrave RE, Tual-Chalot S, Davison BJ, Greally E, Santibanez-Koref M, Schneider JE, Blamire AM, Arthur HM. 2016. Using MRI to predict future adverse cardiac remodelling in a male mouse model of myocardial infarction. Int J Cardiol Heart Vasc, 11 pp. 29-34. | Show Abstract | Read more

BACKGROUND: Mice are frequently used in research to examine outcomes of myocardial infarction (MI) and to investigate therapeutic interventions at an early pre-clinical stage. The MI model is generated by surgically occluding a major coronary artery, but natural variation in murine coronary anatomy can generate variable outcomes that will inevitably affect the accuracy of such investigations. The aim of this study was to use MRI to derive the most sensitive early variable that could be used to predict subsequent adverse cardiac remodelling in a male mouse model of MI. METHODS: Using a longitudinal study design, heart structure and function were evaluated using cardiac MRI at one week following surgical MI to generate the early measurements and again at four weeks, when the scar had matured. The primary variables measured at week one were left ventricular volumes at end systole (LV-ESV) and at end diastole (LV-EDV), infarct size, LV-cardiac mass, and ejection fraction (EF). RESULTS: Univariate and multiple regression analyses showed that LV-ESV at one week following MI could be used to accurately predict various parameters of adverse LV remodelling at four weeks post-MI. However, the highest correlation was between LV-ESV at one week following MI and LV-EDV at four weeks (r = 0.99; p < 0.0001), making LV-ESV at one week a valuable predictor variable of future adverse ventricular remodelling after MI. CONCLUSION: Using MRI to determine LV-ESV at an early stage following MI enables a more robust analysis of potential therapeutic interventions to ameliorate adverse cardiac remodelling.

Wech T, Seiberlich N, Schindele A, Grau V, Diffley L, Gyngell ML, Borzì A, Köstler H, Schneider JE. 2016. Development of Real-Time Magnetic Resonance Imaging of Mouse Hearts at 9.4 Tesla--Simulations and First Application. IEEE Trans Med Imaging, 35 (3), pp. 912-920. | Show Abstract | Read more

A novel method for real-time magnetic resonance imaging for the assessment of cardiac function in mice at 9.4 T is proposed. The technique combines a highly undersampled radial gradient echo acquisition with an image reconstruction utilizing both parallel imaging and compressed sensing. Simulations on an in silico phantom were performed to determine the achievable acceleration factor and to optimize regularization parameters. Several parameters characterizing the quality of the reconstructed images (such as spatial and temporal image sharpness or compartment areas) were calculated for this purpose. Subsequently, double-gated segmented cine data as well as non-gated undersampled real-time data using only six projections per timeframe (temporal resolution  ∼ 10 ms) were acquired in a mid-ventricular slice of four normal mouse hearts in vivo. The highly accelerated data sets were then subjected to the introduced reconstruction technique and results were validated against the fully sampled references. Functional parameters obtained from real-time and fully sampled data agreed well with a comparable accuracy for left-ventricular volumes and a slightly larger scatter for mass. This study introduces and validates a real-time cine-MRI technique, which significantly reduces scan time in preclinical cardiac functional imaging and has the potential to investigate mouse models with abnormal heart rhythm.

Teh I, Maguire ML, Schneider JE. 2017. Efficient gradient calibration based on diffusion MRI. Magn Reson Med, 77 (1), pp. 170-179. | Show Abstract | Read more

PURPOSE: To propose a method for calibrating gradient systems and correcting gradient nonlinearities based on diffusion MRI measurements. METHODS: The gradient scaling in x, y, and z were first offset by up to 5% from precalibrated values to simulate a poorly calibrated system. Diffusion MRI data were acquired in a phantom filled with cyclooctane, and corrections for gradient scaling errors and nonlinearity were determined. The calibration was assessed with diffusion tensor imaging and independently validated with high resolution anatomical MRI of a second structured phantom. RESULTS: The errors in apparent diffusion coefficients along orthogonal axes ranged from -9.2% ± 0.4% to + 8.8% ± 0.7% before calibration and -0.5% ± 0.4% to + 0.8% ± 0.3% after calibration. Concurrently, fractional anisotropy decreased from 0.14 ± 0.03 to 0.03 ± 0.01. Errors in geometric measurements in x, y and z ranged from -5.5% to + 4.5% precalibration and were likewise reduced to -0.97% to + 0.23% postcalibration. Image distortions from gradient nonlinearity were markedly reduced. CONCLUSION: Periodic gradient calibration is an integral part of quality assurance in MRI. The proposed approach is both accurate and efficient, can be setup with readily available materials, and improves accuracy in both anatomical and diffusion MRI to within ±1%. Magn Reson Med 77:170-179, 2017. © 2016 The Authors Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine.

Wilson R, McGuire C, Mohun T, DMDD Project. 2016. Deciphering the mechanisms of developmental disorders: phenotype analysis of embryos from mutant mouse lines. Nucleic Acids Res, 44 (D1), pp. D855-D861. | Show Abstract | Read more

The Deciphering the Mechanisms of Developmental Disorders (DMDD) consortium is a research programme set up to identify genes in the mouse, which if mutated (or knocked-out) result in embryonic lethality when homozygous, and initiate the study of why disruption of their function has such profound effects on embryo development and survival. The project uses a combination of comprehensive high resolution 3D imaging and tissue histology to identify abnormalities in embryo and placental structures of embryonic lethal lines. The image data we have collected and the phenotypes scored are freely available through the project website (http://dmdd.org.uk). In this article we describe the web interface to the images that allows the embryo data to be viewed at full resolution in different planes, discuss how to search the database for a phenotype, and our approach to organising the data for an embryo and a mutant line so it is easy to comprehend and intuitive to navigate.

Allen ER, van Diemen P, Yamaguchi Y, Lindemann C, Soilleux E, Rollier C, Hill F, Schneider J, Wyllie DH. 2016. MRI Based Localisation and Quantification of Abscesses following Experimental S. aureus Intravenous Challenge: Application to Vaccine Evaluation. PLoS One, 11 (5), pp. e0154705. | Show Abstract | Read more

PURPOSE: To develop and validate a sensitive and specific method of abscess enumeration and quantification in a preclinical model of Staphylococcus aureus infection. METHODS: S. aureus infected murine kidneys were fixed in paraformaldehyde, impregnated with gadolinium, and embedded in agar blocks, which were subjected to 3D magnetic resonance microscopy on a 9.4T MRI scanner. Image analysis techniques were developed, which could identify and quantify abscesses. The result of this imaging was compared with histological examination. The impact of a S. aureus Sortase A vaccination regime was assessed using the technique. RESULTS: Up to 32 murine kidneys could be imaged in a single MRI run, yielding images with voxels of about 25 μm3. S. aureus abscesses could be readily identified in blinded analyses of the kidneys after 3 days of infection, with low inter-observer variability. Comparison with histological sections shows a striking correlation between the two techniques: all presumptive abscesses identified by MRI were confirmed histologically, and histology identified no abscesses not evident on MRI. In view of this, simulations were performed assuming that both MRI reconstruction, and histology examining all sections of the tissue, were fully sensitive and specific at abscess detection. This simulation showed that MRI provided more sensitive and precise estimates of abscess numbers and volume than histology, unless at least 5 histological sections are taken through the long axis of the kidney. We used the MRI technique described to investigate the impact of a S. aureus Sortase A vaccine. CONCLUSION: Post mortem MRI scanning of large batches of fixed organs has application in the preclinical assessment of S. aureus vaccines.

Levelt E, Mahmod M, Piechnik SK, Ariga R, Francis JM, Rodgers CT, Clarke WT, Sabharwal N, Schneider JE, Karamitsos TD et al. 2016. Relationship Between Left Ventricular Structural and Metabolic Remodeling in Type 2 Diabetes. Diabetes, 65 (1), pp. 44-52. | Show Abstract | Read more

Concentric left ventricular (LV) remodeling is associated with adverse cardiovascular events and is frequently observed in patients with type 2 diabetes mellitus (T2DM). Despite this, the cause of concentric remodeling in diabetes per se is unclear, but it may be related to cardiac steatosis and impaired myocardial energetics. Thus, we investigated the relationship between myocardial metabolic changes and LV remodeling in T2DM. Forty-six nonhypertensive patients with T2DM and 20 matched control subjects underwent cardiovascular magnetic resonance to assess LV remodeling (LV mass-to-LV end diastolic volume ratio), function, tissue characterization before and after contrast using T1 mapping, and (1)H and (31)P magnetic resonance spectroscopy for myocardial triglyceride content (MTG) and phosphocreatine-to-ATP ratio, respectively. When compared with BMI- and blood pressure-matched control subjects, subjects with diabetes were associated with concentric LV remodeling, higher MTG, impaired myocardial energetics, and impaired systolic strain indicating a subtle contractile dysfunction. Importantly, cardiac steatosis independently predicted concentric remodeling and systolic strain. Extracellular volume fraction was unchanged, indicating the absence of fibrosis. In conclusion, cardiac steatosis may contribute to concentric remodeling and contractile dysfunction of the LV in diabetes. Because cardiac steatosis is modifiable, strategies aimed at reducing MTG may be beneficial in reversing concentric remodeling and improving contractile function in the hearts of patients with diabetes.

Banerjee R, Rial B, Holloway CJ, Lewandowski AJ, Robson MD, Osuchukwu C, Schneider JE, Leeson P, Rider OJ, Neubauer S. 2015. Evidence of a Direct Effect of Myocardial Steatosis on LV Hypertrophy and Diastolic Dysfunction in Adult and Adolescent Obesity JACC-CARDIOVASCULAR IMAGING, 8 (12), pp. 1468-1470. | Read more

McClymont D, Teh I, Whittington HJ, Grau V, Schneider JE. 2016. Prospective acceleration of diffusion tensor imaging with compressed sensing using adaptive dictionaries. Magn Reson Med, 76 (1), pp. 248-258. | Show Abstract | Read more

PURPOSE: Diffusion MRI requires acquisition of multiple diffusion-weighted images, resulting in long scan times. Here, we investigate combining compressed sensing and a fast imaging sequence to dramatically reduce acquisition times in cardiac diffusion MRI. METHODS: Fully sampled and prospectively undersampled diffusion tensor imaging data were acquired in five rat hearts at acceleration factors of between two and six using a fast spin echo (FSE) sequence. Images were reconstructed using a compressed sensing framework, enforcing sparsity by means of decomposition by adaptive dictionaries. A tensor was fit to the reconstructed images and fiber tractography was performed. RESULTS: Acceleration factors of up to six were achieved, with a modest increase in root mean square error of mean apparent diffusion coefficient (ADC), fractional anisotropy (FA), and helix angle. At an acceleration factor of six, mean values of ADC and FA were within 2.5% and 5% of the ground truth, respectively. Marginal differences were observed in the fiber tracts. CONCLUSION: We developed a new k-space sampling strategy for acquiring prospectively undersampled diffusion-weighted data, and validated a novel compressed sensing reconstruction algorithm based on adaptive dictionaries. The k-space undersampling and FSE acquisition each reduced acquisition times by up to 6× and 8×, respectively, as compared to fully sampled spin echo imaging. Magn Reson Med 76:248-258, 2016. © 2015 Wiley Periodicals, Inc.

Teh I, Zhou FL, Hubbard Cristinacce PL, Parker GJ, Schneider JE. 2016. Biomimetic phantom for cardiac diffusion MRI. J Magn Reson Imaging, 43 (3), pp. 594-600. | Show Abstract | Read more

PURPOSE: Diffusion magnetic resonance imaging (MRI) is increasingly used to characterize cardiac tissue microstructure, necessitating the use of physiologically relevant phantoms for methods development. Existing phantoms are generally simplistic and mostly simulate diffusion in the brain. Thus, there is a need for phantoms mimicking diffusion in cardiac tissue. MATERIALS AND METHODS: A biomimetic phantom composed of hollow microfibers generated using co-electrospinning was developed to mimic myocardial diffusion properties and fiber and sheet orientations. Diffusion tensor imaging was carried out at monthly intervals over 4 months at 9.4T. 3D fiber tracking was performed using the phantom and compared with fiber tracking in an ex vivo rat heart. RESULTS: The mean apparent diffusion coefficient and fractional anisotropy of the phantom remained stable over the 4-month period, with mean values of 7.53 ± 0.16 × 10(-4) mm(2) /s and 0.388 ± 0.007, respectively. Fiber tracking of the 1st and 3rd eigenvectors generated analogous results to the fiber and sheet-normal direction respectively, found in the left ventricular myocardium. CONCLUSION: A biomimetic phantom simulating diffusion in the heart was designed and built. This could aid development and validation of novel diffusion MRI methods for investigating cardiac microstructure, decrease the number of animals and patients needed for methods development, and improve quality control in longitudinal and multicenter cardiac diffusion MRI studies.

Maguire ML, Geethanath S, Lygate CA, Kodibagkar VD, Schneider JE. 2015. Compressed sensing to accelerate magnetic resonance spectroscopic imaging: Evaluation and application to &lt;sup&gt;23&lt;/sup&gt;Na-imaging of mouse hearts Journal of Cardiovascular Magnetic Resonance, 17 (1), | Show Abstract | Read more

© 2015 Maguire et al. Background: Magnetic Resonance Spectroscopic Imaging (MRSI) has wide applicability for non-invasive biochemical assessment in clinical and pre-clinical applications but suffers from long scan times. Compressed sensing (CS) has been successfully applied to clinical < sup > 1 < /sup > H MRSI, however a detailed evaluation of CS for conventional chemical shift imaging is lacking. Here we evaluate the performance of CS accelerated MRSI, and specifically apply it to accelerate < sup > 23 < /sup > Na-MRSI on mouse hearts in vivo at 9.4 T. Methods: Synthetic phantom data representing a simplified section across a mouse thorax were used to evaluate the fidelity of the CS reconstruction for varying levels of under-sampling, resolution and signal-to-noise ratios (SNR). The amplitude of signals arising from within a compartment, and signal contamination arising from outside the compartment relative to noise-free Fourier-transformed (FT) data were determined. Simulation results were subsequently verified experimentally in phantoms and in three mouse hearts in vivo. Results: CS reconstructed MRSI data are scaled linearly relative to absolute signal intensities from the fully-sampled FT reconstructed case (R < sup > 2 < /sup > > 0.8, p-value < 0.001). Higher acceleration factors resulted in a denoising of the reconstructed spectra, but also in an increased blurring of compartment boundaries, particularly at lower spatial resolutions. Increasing resolution and SNR decreased cross-compartment contamination and yielded signal amplitudes closer to the FT data. Proof-of-concept high-resolution, 3-fold accelerated < sup > 23 < /sup > Na-amplitude maps of murine myocardium could be obtained within ∼23 mins. Conclusions: Relative signal amplitudes (i.e. metabolite ratios) and absolute quantification of metabolite concentrations can be accurately determined with up to 5-fold under-sampled, CS-reconstructed MRSI. Although this work focused on murine cardiac < sup > 23 < /sup > Na-MRSI, the results are equally applicable to other nuclei and tissues (e.g. < sup > 1 < /sup > H MRSI in brain). Significant reduction in MRSI scan time will reduce the burden on the subject, increase scanner throughput, and may open new avenues for (pre-) clinical metabolic studies.

Miller JJ, Lau AZ, Teh I, Schneider JE, Kinchesh P, Smart S, Ball V, Sibson NR, Tyler DJ. 2016. Robust and high resolution hyperpolarized metabolic imaging of the rat heart at 7 T with 3D spectral-spatial EPI. Magn Reson Med, 75 (4), pp. 1515-1524. | Show Abstract | Read more

PURPOSE: Hyperpolarized metabolic imaging has the potential to revolutionize the diagnosis and management of diseases where metabolism is dysregulated, such as heart disease. We investigated the feasibility of imaging rodent myocardial metabolism at high resolution at 7 T. METHODS: We present here a fly-back spectral-spatial radiofrequency pulse that sidestepped maximum gradient strength requirements and enabled high resolution metabolic imaging of the rodent myocardium. A 3D echo-planar imaging readout followed, with centric ordered z-phase encoding. The cardiac gated sequence was used to image metabolism in rodents whose metabolic state had been manipulated by being fasted, fed, or fed and given the pyruvate dehydrogenase kinase inhibitor dichloroacetate. RESULTS: We imaged hyperpolarized metabolites with a spatial resolution of 2×2×3.8 mm(3) and a temporal resolution of 1.8 s in the rat heart at 7 T. Significant differences in myocardial pyruvate dehydrogenase flux were observed between the three groups of animals, concomitant with the known biochemistry. CONCLUSION: The proposed sequence was able to image in vivo metabolism with excellent spatial resolution in the rat heart. The field of view enabled the simultaneous multi-organ acquisition of metabolic information from the rat, which is of great utility for preclinical research in cardiovascular disease. Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance.

Banerjee R, Rial B, Holloway CJ, Lewandowski AJ, Robson MD, Osuchukwu C, Schneider JE, Leeson P, Rider OJ, Neubauer S. 2015. Evidence of a Direct Effect of Myocardial Steatosis on LV Hypertrophy and Diastolic Dysfunction in Adult and Adolescent Obesity. JACC Cardiovasc Imaging, 8 (12), pp. 1468-1470. | Read more

Maguire ML, Geethanath S, Lygate CA, Kodibagkar VD, Schneider JE. 2015. Compressed sensing to accelerate magnetic resonance spectroscopic imaging: evaluation and application to 23Na-imaging of mouse hearts. J Cardiovasc Magn Reson, 17 (1), pp. 45. | Show Abstract | Read more

BACKGROUND: Magnetic Resonance Spectroscopic Imaging (MRSI) has wide applicability for non-invasive biochemical assessment in clinical and pre-clinical applications but suffers from long scan times. Compressed sensing (CS) has been successfully applied to clinical 1H MRSI, however a detailed evaluation of CS for conventional chemical shift imaging is lacking. Here we evaluate the performance of CS accelerated MRSI, and specifically apply it to accelerate 23Na-MRSI on mouse hearts in vivo at 9.4 T. METHODS: Synthetic phantom data representing a simplified section across a mouse thorax were used to evaluate the fidelity of the CS reconstruction for varying levels of under-sampling, resolution and signal-to-noise ratios (SNR). The amplitude of signals arising from within a compartment, and signal contamination arising from outside the compartment relative to noise-free Fourier-transformed (FT) data were determined. Simulation results were subsequently verified experimentally in phantoms and in three mouse hearts in vivo. RESULTS: CS reconstructed MRSI data are scaled linearly relative to absolute signal intensities from the fully-sampled FT reconstructed case (R(2) > 0.8, p-value < 0.001). Higher acceleration factors resulted in a denoising of the reconstructed spectra, but also in an increased blurring of compartment boundaries, particularly at lower spatial resolutions. Increasing resolution and SNR decreased cross-compartment contamination and yielded signal amplitudes closer to the FT data. Proof-of-concept high-resolution, 3-fold accelerated 23Na-amplitude maps of murine myocardium could be obtained within ~23 mins. CONCLUSIONS: Relative signal amplitudes (i.e. metabolite ratios) and absolute quantification of metabolite concentrations can be accurately determined with up to 5-fold under-sampled, CS-reconstructed MRSI. Although this work focused on murine cardiac 23Na-MRSI, the results are equally applicable to other nuclei and tissues (e.g., 1H MRSI in brain). Significant reduction in MRSI scan time will reduce the burden on the subject, increase scanner throughput, and may open new avenues for (pre-) clinical metabolic studies.

Lohezic M, Bollensdorff C, Korn M, Lanz T, Grau V, Kohl P, Schneider JE. 2015. Optimized radiofrequency coil setup for MR examination of living isolated rat hearts in a horizontal 9.4T magnet Magnetic Resonance in Medicine, 73 (6), pp. 2398-2405. | Show Abstract | Read more

© 2014 The Authors. Purpose (i) To optimize an MR-compatible organ perfusion setup for the nondestructive investigation of isolated rat hearts by placing the radiofrequency (RF) coil inside the perfusion chamber; (ii) to characterize the benefit of this system for diffusion tensor imaging and proton ( < sup > 1 < /sup > H-) MR spectroscopy. Methods Coil quality assessment was conducted both on the bench, and in the magnet. The benefit of the new RF-coil was quantified by measuring signal-to-noise ratio (SNR), accuracy, and precision of diffusion tensor imaging/error in metabolite amplitude estimation, and compared to an RF-coil placed externally to the perfusion chamber. Results The new design provided a 59% gain in signal-to-noise ratio on a fixed rat heart compared to using an external resonator, which found reflection in an improvement of living heart data quality, compared to previous external resonator studies. This resulted in 14-29% improvement in accuracy and precision of diffusion tensor imaging. The Cramer- Rao lower bounds for metabolite amplitude estimations were up to 5-fold smaller. Conclusion Optimization of MR-compatible perfusion equipment advances the study of rat hearts with improved signal-to-noise ratio performance, and thus improved accuracy/precision. Magn Reson Med 73:2398-2405, 2015.

Faller KM, McAndrew DJ, Schneider JE, Lygate CA. 2015. Refinement of analgesia following thoracotomy and experimental myocardial infarction using the Mouse Grimace Scale. Exp Physiol, 100 (2), pp. 164-172. | Show Abstract | Read more

NEW FINDINGS: What is the central question of this study? There is an ethical imperative to optimize analgesia protocols for laboratory animals, but this is impeded by our inability to recognize pain reliably. We examined whether the Mouse Grimace Scale (MGS) provides benefits over a standard welfare scoring system for identifying a low level of pain in the frequently used murine surgical model of myocardial infarction. What is the main finding and its importance? Low-level pain, responsive to analgesia, was detected by MGS but not standard methods. In this model, most of the pain is attributable to the thoracotomy, excepted in mice with very large infarcts. This approach represents a model for assessing postsurgical analgesia in rodents. The Mouse Grimace Scale (MGS) was developed for assessing pain severity, but the general applicability to complex postsurgical pain has not been established. We sought to determine whether the MGS provides benefits over and above a standard welfare scoring system for identifying pain in mice following experimental myocardial infarction. Female C57BL/6J mice (n = 60), anaesthetized with isoflurane, were subjected to thoracotomy with ligation of a coronary artery or sham procedure. A single s.c. dose of buprenorphine (1.1 mg kg(-1) ) was given at the time of surgery and pain assessed at 24 h by MGS and a procedure-specific welfare scoring system. In some animals, a second dose of 0.6 mg kg(-1) buprenorphine was given and pain assessment repeated after 30 min. The MGS was scored from multiple photographs by two independent blinded observers with good correlation (r = 0.98). Using the average MGS score of both observers, we identified a subset of mice with low scores that were not considered to be in pain by the welfare scoring system or by single observer MGS. These mice showed a significant improvement with additional analgesia, suggesting that this low-level pain is real. Pain attributable to the myocardial injury, as opposed to thoracotomy, persisted at 24 h only in mice with large infarcts >40%. In conclusion, the use of a multi-observer, post hoc version of the MGS is a sensitive tool to assess the efficacy of postsurgical analgesic protocols. Following surgical induction of myocardial infarction, we identified a significant proportion of mice that were in low-level pain at 24 h that were not identified by other assessment methods.

Burton RA, Lee P, Casero R, Garny A, Siedlecka U, Schneider JE, Kohl P, Grau V. 2014. Three-dimensional histology: tools and application to quantitative assessment of cell-type distribution in rabbit heart. Europace, 16 Suppl 4 (suppl 4), pp. iv86-iv95. | Show Abstract | Read more

AIMS: Cardiac histo-anatomical organization is a major determinant of function. Changes in tissue structure are a relevant factor in normal and disease development, and form targets of therapeutic interventions. The purpose of this study was to test tools aimed to allow quantitative assessment of cell-type distribution from large histology and magnetic resonance imaging- (MRI) based datasets. METHODS AND RESULTS: Rabbit heart fixation during cardioplegic arrest and MRI were followed by serial sectioning of the whole heart and light-microscopic imaging of trichrome-stained tissue. Segmentation techniques developed specifically for this project were applied to segment myocardial tissue in the MRI and histology datasets. In addition, histology slices were segmented into myocytes, connective tissue, and undefined. A bounding surface, containing the whole heart, was established for both MRI and histology. Volumes contained in the bounding surface (called 'anatomical volume'), as well as that identified as containing any of the above tissue categories (called 'morphological volume'), were calculated. The anatomical volume was 7.8 cm(3) in MRI, and this reduced to 4.9 cm(3) after histological processing, representing an 'anatomical' shrinkage by 37.2%. The morphological volume decreased by 48% between MRI and histology, highlighting the presence of additional tissue-level shrinkage (e.g. an increase in interstitial cleft space). The ratio of pixels classified as containing myocytes to pixels identified as non-myocytes was roughly 6:1 (61.6 vs. 9.8%; the remaining fraction of 28.6% was 'undefined'). CONCLUSION: Qualitative and quantitative differentiation between myocytes and connective tissue, using state-of-the-art high-resolution serial histology techniques, allows identification of cell-type distribution in whole-heart datasets. Comparison with MRI illustrates a pronounced reduction in anatomical and morphological volumes during histology processing.

Lamata P, Casero R, Carapella V, Niederer SA, Bishop MJ, Schneider JE, Kohl P, Grau V. 2014. Images as drivers of progress in cardiac computational modelling. Prog Biophys Mol Biol, 115 (2-3), pp. 198-212. | Show Abstract | Read more

Computational models have become a fundamental tool in cardiac research. Models are evolving to cover multiple scales and physical mechanisms. They are moving towards mechanistic descriptions of personalised structure and function, including effects of natural variability. These developments are underpinned to a large extent by advances in imaging technologies. This article reviews how novel imaging technologies, or the innovative use and extension of established ones, integrate with computational models and drive novel insights into cardiac biophysics. In terms of structural characterization, we discuss how imaging is allowing a wide range of scales to be considered, from cellular levels to whole organs. We analyse how the evolution from structural to functional imaging is opening new avenues for computational models, and in this respect we review methods for measurement of electrical activity, mechanics and flow. Finally, we consider ways in which combined imaging and modelling research is likely to continue advancing cardiac research, and identify some of the main challenges that remain to be solved.

Lohezic M, Teh I, Bollensdorff C, Peyronnet R, Hales PW, Grau V, Kohl P, Schneider JE. 2014. Interrogation of living myocardium in multiple static deformation states with diffusion tensor and diffusion spectrum imaging. Prog Biophys Mol Biol, 115 (2-3), pp. 213-225. | Show Abstract | Read more

Diffusion tensor magnetic resonance imaging (MRI) reveals valuable insights into tissue histo-anatomy and microstructure, and has steadily gained traction in the cardiac community. Its wider use in small animal cardiac imaging in vivo has been constrained by its extreme sensitivity to motion, exaggerated by the high heart rates usually seen in rodents. Imaging of the isolated heart eliminates respiratory motion and, if conducted on arrested hearts, cardiac pulsation. This serves as an important intermediate step for basic and translational studies. However, investigating the micro-structural basis of cardiac deformation in the same heart requires observations in different deformation states. Here, we illustrate the imaging of isolated rat hearts in three mechanical states mimicking diastole (cardioplegic arrest), left-ventricular (LV) volume overload (cardioplegic arrest plus LV balloon inflation), and peak systole (lithium-induced contracture). An optimised MRI-compatible Langendorff perfusion setup with the radio-frequency (RF) coil integrated into the wet chamber was developed for use in a 9.4T horizontal bore scanner. Signal-to-noise ratio improved significantly, by 75% compared to a previous design with external RF coil, and stability tests showed no significant changes in mean T1, T2 or LV wall thickness over a 170 min period. In contracture, we observed a significant reduction in mean fractional anisotropy from 0.32 ± 0.02 to 0.28 ± 0.02, as well as a significant rightward shift in helix angles with a decrease in the proportion of left-handed fibres, as referring to the locally prevailing cell orientation in the heart, from 24.9% to 23.3%, and an increase in the proportion of right-handed fibres from 25.5% to 28.4%. LV overload, in contrast, gave rise to a decrease in the proportion of left-handed fibres from 24.9% to 21.4% and an increase in the proportion of right-handed fibres from 25.5% to 26.0%. The modified perfusion and coil setup offers better performance and control over cardiac contraction states. We subsequently performed high-resolution diffusion spectrum imaging (DSI) and 3D whole heart fibre tracking in fixed ex vivo rat hearts in slack state and contracture. As a model-free method, DSI augmented the measurements of water diffusion by also informing on multiple intra-voxel diffusion orientations and non-Gaussian diffusion. This enabled us to identify the transition from right- to left-handed fibres from the subendocardium to the subepicardium, as well as voxels in apical regions that were traversed by multiple fibres. We observed that both the mean generalised fractional anisotropy and mean kurtosis were lower in hearts in contracture compared to the slack state, by 23% and 9.3%, respectively. While its heavy acquisition burden currently limits the application of DSI in vivo, ongoing work in acceleration techniques may enable its use in live animals and patients. This would provide access to the as yet unexplored dimension of non-Gaussian diffusion that could serve as a highly sensitive marker of cardiac micro-structural integrity.

Nordbø O, Lamata P, Land S, Niederer S, Aronsen JM, Louch WE, Sjaastad I, Martens H, Gjuvsland AB, Tøndel K et al. 2014. A computational pipeline for quantification of mouse myocardial stiffness parameters. Comput Biol Med, 53 pp. 65-75. | Show Abstract | Read more

The mouse is an important model for theoretical-experimental cardiac research, and biophysically based whole organ models of the mouse heart are now within reach. However, the passive material properties of mouse myocardium have not been much studied. We present an experimental setup and associated computational pipeline to quantify these stiffness properties. A mouse heart was excised and the left ventricle experimentally inflated from 0 to 1.44kPa in eleven steps, and the resulting deformation was estimated by echocardiography and speckle tracking. An in silico counterpart to this experiment was built using finite element methods and data on ventricular tissue microstructure from diffusion tensor MRI. This model assumed a hyperelastic, transversely isotropic material law to describe the force-deformation relationship, and was simulated for many parameter scenarios, covering the relevant range of parameter space. To identify well-fitting parameter scenarios, we compared experimental and simulated outcomes across the whole range of pressures, based partly on gross phenotypes (volume, elastic energy, and short- and long-axis diameter), and partly on node positions in the geometrical mesh. This identified a narrow region of experimentally compatible values of the material parameters. Estimation turned out to be more precise when based on changes in gross phenotypes, compared to the prevailing practice of using displacements of the material points. We conclude that the presented experimental setup and computational pipeline is a viable method that deserves wider application.

Lamata P, Casero R, Carapella V, Niederer SA, Bishop MJ, Schneider JE, Kohl P, Grau V. 2014. Images as drivers of progress in cardiac computational modelling Progress in Biophysics and Molecular Biology, 115 (2-3), pp. 198-212. | Show Abstract | Read more

© 2014 Elsevier Ltd. Computational models have become a fundamental tool in cardiac research. Models are evolving to cover multiple scales and physical mechanisms. They are moving towards mechanistic descriptions of personalised structure and function, including effects of natural variability. These developments are underpinned to a large extent by advances in imaging technologies. This article reviews how novel imaging technologies, or the innovative use and extension of established ones, integrate with computational models and drive novel insights into cardiac biophysics. In terms of structural characterization, we discuss how imaging is allowing a wide range of scales to be considered, from cellular levels to whole organs. We analyse how the evolution from structural to functional imaging is opening new avenues for computational models, and in this respect we review methods for measurement of electrical activity, mechanics and flow. Finally, we consider ways in which combined imaging and modelling research is likely to continue advancing cardiac research, and identify some of the main challenges that remain to be solved.

Lohezic M, Teh I, Bollensdorff C, Peyronnet R, Hales PW, Grau V, Kohl P, Schneider JE. 2014. Interrogation of living myocardium in multiple static deformation states with diffusion tensor and diffusion spectrum imaging Progress in Biophysics and Molecular Biology, 115 (2-3), pp. 213-225. | Show Abstract | Read more

© 2014 The Authors. Diffusion tensor magnetic resonance imaging (MRI) reveals valuable insights into tissue histo-anatomy and microstructure, and has steadily gained traction in the cardiac community. Its wider use in small animal cardiac imaging invivo has been constrained by its extreme sensitivity to motion, exaggerated by the high heart rates usually seen in rodents. Imaging of the isolated heart eliminates respiratory motion and, if conducted on arrested hearts, cardiac pulsation. This serves as an important intermediate step for basic and translational studies. However, investigating the micro-structural basis of cardiac deformation in the same heart requires observations in different deformation states. Here, we illustrate the imaging of isolated rat hearts in three mechanical states mimicking diastole (cardioplegic arrest), left-ventricular (LV) volume overload (cardioplegic arrest plus LV balloon inflation), and peak systole (lithium-induced contracture). An optimised MRI-compatible Langendorff perfusion setup with the radio-frequency (RF) coil integrated into the wet chamber was developed for use in a 9.4T horizontal bore scanner. Signal-to-noise ratio improved significantly, by 75% compared to a previous design with external RF coil, and stability tests showed no significant changes in mean T 1 , T 2 or LV wall thickness over a 170min period. In contracture, we observed a significant reduction in mean fractional anisotropy from 0.32±0.02 to 0.28±0.02, as well as a significant rightward shift in helix angles with a decrease in the proportion of left-handed fibres, as referring to the locally prevailing cell orientation in the heart, from 24.9% to 23.3%, and an increase in the proportion of right-handed fibres from 25.5% to 28.4%. LV overload, in contrast, gave rise to a decrease in the proportion of left-handed fibres from 24.9% to 21.4% and an increase in the proportion of right-handed fibres from 25.5% to 26.0%. The modified perfusion and coil setup offers better performance and control over cardiac contraction states. We subsequently performed high-resolution diffusion spectrum imaging (DSI) and 3D whole heart fibre tracking in fixed exvivo rat hearts in slack state and contracture. As a model-free method, DSI augmented the measurements of water diffusion by also informing on multiple intra-voxel diffusion orientations and non-Gaussian diffusion. This enabled us to identify the transition from right- to left-handed fibres from the subendocardium to the subepicardium, as well as voxels in apical regions that were traversed by multiple fibres. We observed that both the mean generalised fractional anisotropy and mean kurtosis were lower in hearts in contracture compared to the slack state, by 23% and 9.3%, respectively. While its heavy acquisition burden currently limits the application of DSI invivo, ongoing work in acceleration techniques may enable its use in live animals and patients. This would provide access to the as yet unexplored dimension of non-Gaussian diffusion that could serve as a highly sensitive marker of cardiac micro-structural integrity.

Lohezic M, Bollensdorff C, Korn M, Lanz T, Grau V, Kohl P, Schneider JE. 2015. Optimized radiofrequency coil setup for MR examination of living isolated rat hearts in a horizontal 9.4T magnet. Magn Reson Med, 73 (6), pp. 2398-2405. | Show Abstract | Read more

PURPOSE: (i) To optimize an MR-compatible organ perfusion setup for the nondestructive investigation of isolated rat hearts by placing the radiofrequency (RF) coil inside the perfusion chamber; (ii) to characterize the benefit of this system for diffusion tensor imaging and proton ((1) H-) MR spectroscopy. METHODS: Coil quality assessment was conducted both on the bench, and in the magnet. The benefit of the new RF-coil was quantified by measuring signal-to-noise ratio (SNR), accuracy, and precision of diffusion tensor imaging/error in metabolite amplitude estimation, and compared to an RF-coil placed externally to the perfusion chamber. RESULTS: The new design provided a 59% gain in signal-to-noise ratio on a fixed rat heart compared to using an external resonator, which found reflection in an improvement of living heart data quality, compared to previous external resonator studies. This resulted in 14-29% improvement in accuracy and precision of diffusion tensor imaging. The Cramer-Rao lower bounds for metabolite amplitude estimations were up to 5-fold smaller. CONCLUSION: Optimization of MR-compatible perfusion equipment advances the study of rat hearts with improved signal-to-noise ratio performance, and thus improved accuracy/precision.

Levelt E, Ntusi N, Mahmod M, Wainwright C, Piechnik S, Francis J, Davis A, Schneider J, Leeson P, Karamitsos T et al. 2014. 127 Early manifestations of diabetic cardiomyopathy assessed by cardiac magnetic resonance imaging and spectroscopy. Heart, 100 Suppl 3 (Suppl 3), pp. A73-A74. | Show Abstract | Read more

Large, population based studies have shown that diabetes mellitus (DM) of long durationis associated with increased incidence of heart failure, independent of underlying hypertension and coronary artery disease. Cardiovascular magnetic resonance imaging (CMR) and magnetic resonance spectroscopy (MRS) provide comprehensive, non-invasive, multiparametric measures of the functional, structural and metabolic status of the heart. We used magnetic resonance imaging and spectroscopy to determine whether subclinical functional, structural and metabolic alterations occur in a patient cohort of early-onset, type 2 DM.

Fan LM, Douglas G, Bendall JK, McNeill E, Crabtree MJ, Hale AB, Mai A, Li JM, McAteer MA, Schneider JE et al. 2014. Endothelial cell-specific reactive oxygen species production increases susceptibility to aortic dissection. Circulation, 129 (25), pp. 2661-2672. | Show Abstract | Read more

BACKGROUND: Increased production of reactive oxygen species (ROS) throughout the vascular wall is a feature of cardiovascular disease states, but therapeutic strategies remain limited by our incomplete understanding of the role and contribution of specific vascular cell ROS to disease pathogenesis. To investigate the specific role of endothelial cell (EC) ROS in the development of structural vascular disease, we generated a mouse model of endothelium-specific Nox2 overexpression and tested the susceptibility to aortic dissection after angiotensin II (Ang II) infusion. METHODS AND RESULTS: A specific increase in endothelial ROS production in Nox2 transgenic mice was sufficient to cause Ang II-mediated aortic dissection, which was never observed in wild-type mice. Nox2 transgenic aortas had increased endothelial ROS production, endothelial vascular cell adhesion molecule-1 expression, matrix metalloproteinase activity, and CD45(+) inflammatory cell infiltration. Conditioned media from Nox2 transgenic ECs induced greater Erk1/2 phosphorylation in vascular smooth muscle cells compared with wild-type controls through secreted cyclophilin A (CypA). Nox2 transgenic ECs (but not vascular smooth muscle cells) and aortas had greater secretion of CypA both at baseline and in response to Ang II stimulation. Knockdown of CypA in ECs abolished the increase in vascular smooth muscle cell Erk1/2 phosphorylation conferred by EC conditioned media, and preincubation with CypA augmented Ang II-induced vascular smooth muscle cell ROS production. CONCLUSIONS: These findings demonstrate a pivotal role for EC-derived ROS in the determination of the susceptibility of the aortic wall to Ang II-mediated aortic dissection. ROS-dependent CypA secretion by ECs is an important signaling mechanism through which EC ROS regulate susceptibility of structural components of the aortic wall to aortic dissection.

Secchi F, Cannao P, Pluchinotta F, Butera G, Carminati M, Sardanelli F, Lombardi M, Monney P, Piccini D, Rutz T et al. 2014. These abstracts have been selected for VIEWING only as ePosters and in print. ePosters will be available on Screen A & B throughout the meeting, Print Posters at the times indicated below. Please refer to the PROGRAM for more details. Eur Heart J Cardiovasc Imaging, 15 Suppl 1 (suppl 1), pp. i12-i33. | Read more

Constantinides C, Carr CA, Schneider JE. 2015. Recent Advances in Image-Based Stem-Cell Labeling and Tracking, and Scaffold-Based Organ Development in Cardiovascular Disease Recent Patents on Medical Imaging, 4 (2), pp. 110-126. | Show Abstract | Read more

© 2014 Bentham Science Publishers. Myocardial infarction (MI)and heart failure (HF)are leading causes of mortality and morbidity in the Western World. Therapeutic approaches using interventional cardiology and bioengineering techniques have thus far focused on either salvaging viable tissue post-infarction or preserving cardiac function in the failing myocardium. Regenerative medicine on the other hand, attempts to renew damaged tissue and enhance cardiac functional performance. Tremendous advances have been made in this field since the introduction and ethical approval for use of stem-cells (SC)and relevant technologies in pre-clinical and clinical practice. While study outcomes are still ambivalent on the potential translational impact of SCs, renewed hope has arisen since the introduction of induced pluripotent stem-cells (iPS)and the prospect of intact organ development and transplantation. The aim of this work is to review recent discoveries and the patent landscape employing stem-cell engineering, labeling and image-based monitoring strategies, their use in bioreactors and constructions of enriched bio-artificial membranes, as well as the potential role in artificial organ development and transplantation, with relevance to anticipated impact in pre-clinical screening and widespread clinical use.

Aksentijević D, Zervou S, Faller KM, McAndrew DJ, Schneider JE, Neubauer S, Lygate CA. 2014. Myocardial creatine levels do not influence response to acute oxidative stress in isolated perfused heart. PLoS One, 9 (10), pp. e109021. | Show Abstract | Read more

BACKGROUND: Multiple studies suggest creatine mediates anti-oxidant activity in addition to its established role in cellular energy metabolism. The functional significance for the heart has yet to be established, but antioxidant activity could contribute to the cardioprotective effect of creatine in ischaemia/reperfusion injury. OBJECTIVES: To determine whether intracellular creatine levels influence responses to acute reactive oxygen species (ROS) exposure in the intact beating heart. We hypothesised that mice with elevated creatine due to over-expression of the creatine transporter (CrT-OE) would be relatively protected, while mice with creatine-deficiency (GAMT KO) would fare worse. METHODS AND RESULTS: CrT-OE mice were pre-selected for creatine levels 20-100% above wild-type using in vivo (1)H-MRS. Hearts were perfused in isovolumic Langendorff mode and cardiac function monitored throughout. After 20 min equilibration, hearts were perfused with either H2O2 0.5 µM (30 min), or the anti-neoplastic drug doxorubicin 15 µM (100 min). Protein carbonylation, creatine kinase isoenzyme activities and phospho-PKCδ expression were quantified in perfused hearts as markers of oxidative damage and apoptotic signalling. Wild-type hearts responded to ROS challenge with a profound decline in contractile function that was ameliorated by co-administration of catalase or dexrazoxane as positive controls. In contrast, the functional deterioration in CrT-OE and GAMT KO hearts was indistinguishable from wild-type controls, as was the extent of oxidative damage and apoptosis. Exogenous creatine supplementation also failed to protect hearts from doxorubicin-induced dysfunction. CONCLUSIONS: Intracellular creatine levels do not influence the response to acute ROS challenge in the intact beating heart, arguing against creatine exerting (patho-)physiologically relevant anti-oxidant activity.

Carapella V, Bordas R, Pathmanathan P, Lohezic M, Schneider JE, Kohl P, Burrage K, Grau V. 2014. Quantitative study of the effect of tissue microstructure on contraction in a computational model of rat left ventricle. PLoS One, 9 (4), pp. e92792. | Show Abstract | Read more

Tissue microstructure, in particular the alignment of myocytes (fibre direction) and their lateral organisation into sheets, is fundamental to cardiac function. We studied the effect of microstructure on contraction in a computational model of rat left ventricular electromechanics. Different fibre models, globally rule-based or locally optimised to DT-MRI data, were compared, in order to understand whether a subject-specific fibre model would enhance the predictive power of our model with respect to the global ones. We also studied the impact of sheets on ventricular deformation by comparing: (a) a transversely isotropic versus an orthotropic material law and (b) a linear model with a bimodal model of sheet transmural variation. We estimated ejection fraction, wall thickening and base-to-apex shortening and compared them with measures from cine-MRI. We also evaluated Lagrangian strains as local metrics of cardiac deformation. Our results show that the subject-specific fibre model provides little improvement in the metric predictions with respect to global fibre models while material orthotropy allows closer agreement with measures than transverse isotropy. Nonetheless, the impact of sheets in our model is smaller than that of fibres. We conclude that further investigation of the modelling of sheet dynamics is necessary to fully understand the impact of tissue structure on cardiac deformation.

Banerjee R, Pavlides M, Tunnicliffe EM, Piechnik SK, Sarania N, Philips R, Collier JD, Booth JC, Schneider JE, Wang LM et al. 2014. Multiparametric magnetic resonance for the non-invasive diagnosis of liver disease. J Hepatol, 60 (1), pp. 69-77. | Show Abstract | Read more

BACKGROUND & AIMS: With the increasing prevalence of liver disease worldwide, there is an urgent clinical need for reliable methods to diagnose and stage liver pathology. Liver biopsy, the current gold standard, is invasive and limited by sampling and observer dependent variability. In this study, we aimed to assess the diagnostic accuracy of a novel magnetic resonance protocol for liver tissue characterisation. METHODS: We conducted a prospective study comparing our magnetic resonance technique against liver biopsy. The individual components of the scanning protocol were T1 mapping, proton spectroscopy and T2* mapping, which quantified liver fibrosis, steatosis and haemosiderosis, respectively. Unselected adult patients referred for liver biopsy as part of their routine care were recruited. Scans performed prior to liver biopsy were analysed by physicians blinded to the histology results. The associations between magnetic resonance and histology variables were assessed. Receiver-operating characteristic analyses were also carried out. RESULTS: Paired magnetic resonance and biopsy data were obtained in 79 patients. Magnetic resonance measures correlated strongly with histology (r(s)=0.68 p<0.0001 for fibrosis; r(s)=0.89 p<0.001 for steatosis; r(s)=-0.69 p<0.0001 for haemosiderosis). The area under the receiver operating characteristic curve was 0.94, 0.93, and 0.94 for the diagnosis of any degree of fibrosis, steatosis and haemosiderosis respectively. CONCLUSION: The novel scanning method described here provides high diagnostic accuracy for the assessment of liver fibrosis, steatosis and haemosiderosis and could potentially replace liver biopsy for many indications. This is the first demonstration of a non-invasive test to differentiate early stages of fibrosis from normal liver.

Lygate CA, Faller KM, Medway DJ, Zervou S, Choe C-U, Isbrandt D, Schneider JE, Neubauer S. 2013. Chronic Creatine-deficiency is not the Underlying Cause of Cardiac Dysfunction in Mice Lacking the Creatine Biosynthetic Enzyme Arginine: glycine Amidinotransferase CIRCULATION, 128 (22),

Espe EK, Aronsen JM, Skårdal K, Schneider JE, Zhang L, Sjaastad I. 2013. Novel insight into the detailed myocardial motion and deformation of the rodent heart using high-resolution phase contrast cardiovascular magnetic resonance. J Cardiovasc Magn Reson, 15 (1), pp. 82. | Show Abstract | Read more

BACKGROUND: Phase contrast velocimetry cardiovascular magnetic resonance (PC-CMR) is a powerful and versatile tool allowing assessment of in vivo motion of the myocardium. However, PC-CMR is sensitive to motion related artifacts causing errors that are geometrically systematic, rendering regional analysis of myocardial function challenging. The objective of this study was to establish an optimized PC-CMR method able to provide novel insight in the complex regional motion and strain of the rodent myocardium, and provide a proof-of-concept in normal and diseased rat hearts with higher temporal and spatial resolution than previously reported. METHODS: A PC-CMR protocol optimized for assessing the motion and deformation of the myocardium in rats with high spatiotemporal resolution was established, and ten animals with different degree of cardiac dysfunction underwent examination and served as proof-of-concept. Global and regional myocardial velocities and circumferential strain were calculated, and the results were compared to five control animals. Furthermore, the global strain measurements were validated against speckle-tracking echocardiography, and inter- and intrastudy variability of the protocol were evaluated. RESULTS: The presented method allows assessment of regional myocardial function in rats with high level of detail; temporal resolution was 3.2 ms, and analysis was done using 32 circumferential segments. In the dysfunctional hearts, global and regional function were distinctly altered, including reduced global peak values, increased regional heterogeneity and increased index of dyssynchrony. Strain derived from the PC-CMR data was in excellent agreement with echocardiography (r = 0.95, p < 0.001; limits-of-agreement -0.02 ± 3.92%strain), and intra- and interstudy variability were low for both velocity and strain (limits-of-agreement, radial motion: 0.01 ± 0.32 cm/s and -0.06 ± 0.75 cm/s; circumferential strain: -0.16 ± 0.89%strain and -0.71 ± 1.67%strain, for intra- and interstudy, respectively). CONCLUSION: We demonstrate, for the first time, that PC-CMR enables high-resolution evaluation of in vivo circumferential strain in addition to myocardial motion of the rat heart. In combination with the superior geometric robustness of CMR, this ultimately provides a tool for longitudinal studies of regional function in rodents with high level of detail.

Faller KME, Lygate CA, Neubauer S, Schneider JE. 2013. 1H-MR spectroscopy for analysis of cardiac lipid and creatine metabolism Heart Failure Reviews, 18 (5), pp. 657-668. | Show Abstract | Read more

Magnetic resonance spectroscopy (MRS) is the only non-invasive, non-radiation-based technique for investigating the metabolism of living tissue. MRS of protons ( 1 H-MRS), which provides the highest sensitivity of all MR-visible nuclei, is a method capable of detecting and quantifying specific cardiac biomolecules, such as lipids and creatine in normal and diseased hearts in both animal models and humans. This can be used to study mechanisms of heart failure development in a longitudinal manner, for example, the potential contribution of myocardial lipid accumulation in the context of ageing and obesity. Similarly, quantifying creatine levels provides insight into the energy storage and buffering capacity in the heart. Creatine depletion is consistently observed in heart failure independent of aetiology, but its contribution to pathophysiology remains a matter of debate. These and other questions can in theory be answered with cardiac MRS, but fundamental technical challenges have limited its use. The metabolites studied with MRS are much lower concentration than water protons, requiring methods to suppress the dominant water signal and resulting in larger voxel sizes and longer scan times compared to MRI. However, recent technical advances in MR hardware and software have facilitated the application of 1 H-MRS in humans and animal models of heart disease as detailed in this review. © 2012 The Author(s).

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Holloway CJ, Ntusi N, Suttie J, Mahmod M, Wainwright E, Clutton G, Hancock G, Beak P, Tajar A, Piechnik SK et al. 2013. Comprehensive cardiac magnetic resonance imaging and spectroscopy reveal a high burden of myocardial disease in HIV patients Circulation, 128 (8), pp. 814-822. | Show Abstract | Read more

BACKGROUND - : HIV infection continues to be endemic worldwide. Although treatments are successful, it remains controversial whether patients receiving optimal therapy have structural, functional, or biochemical cardiac abnormalities that may underlie their increased cardiac morbidity and mortality. The purpose of this study was to characterize myocardial abnormalities in a contemporary group of HIV-infected individuals undergoing combination antiretroviral therapy. METHODS AND RESULTS - : Volunteers with HIV who were undergoing combination antiretroviral therapy and age-matched control subjects without a history of cardiovascular disease underwent cardiac magnetic resonance imaging and spectroscopy for the determination of cardiac function, myocardial fibrosis, and myocardial lipid content. A total of 129 participants were included in this analysis. Compared with age-matched control subjects (n=39; 30.23%), HIV-infected subjects undergoing combination antiretroviral therapy (n=90; 69.77%) had 47% higher median myocardial lipid levels (P < 0.003) and 74% higher median plasma triglyceride levels (both P < 0.001). Myocardial fibrosis, predominantly in the basal inferolateral wall of the left ventricle, was observed in 76% of HIV-infected subjects compared with 13% of control subjects (P < 0.001). Peak myocardial systolic and diastolic longitudinal strain were also lower in HIV-infected individuals than in control subjects and remained statistically significant after adjustment for available confounders. CONCLUSIONS - : Comprehensive cardiac imaging revealed cardiac steatosis, alterations in cardiac function, and a high prevalence of myocardial fibrosis in a contemporary group of asymptomatic HIV-infected subjects undergoing combination antiretroviral therapy. Cardiac steatosis and fibrosis may underlie cardiac dysfunction and increased cardiovascular morbidity and mortality in subjects with HIV. © 2013 American Heart Association, Inc.

Mahmod M, Bull S, Suttie JJ, Pal N, Holloway C, Dass S, Myerson SG, Schneider JE, De Silva R, Petrou M et al. 2013. Myocardial steatosis and left ventricular contractile dysfunction in patients with severe aortic stenosis. Circ Cardiovasc Imaging, 6 (5), pp. 808-816. | Show Abstract | Read more

BACKGROUND: Aortic stenosis (AS) leads to left ventricular (LV) hypertrophy and dysfunction. We hypothesized that cardiac steatosis is involved in the pathophysiology and also assessed whether it is reversible after aortic valve replacement. METHODS AND RESULTS: Thirty-nine patients with severe AS (symptomatic=25, asymptomatic=14) with normal LV ejection fraction and no significant coronary artery disease and 20 age- and sex-matched healthy controls underwent cardiac 1H-magnetic resonance spectroscopy and imaging for the determination of steatosis (myocardial triglyceride content) and cardiac function, including circumferential strain (measured by magnetic resonance tagging). Strain was lower in both symptomatic and asymptomatic AS (-16.4 ± 2.5% and -18.1 ± 2.9%, respectively, versus controls -20.7 ± 2.0%, both P<0.05). Myocardial steatosis was found in both symptomatic and asymptomatic patients with AS (0.89 ± 0.42% in symptomatic AS; 0.75 ± 0.36% in asymptomatic AS versus controls 0.45 ± 0.17, both P<0.05). Importantly, multivariable analysis indicated that steatosis was an independent correlate of impaired LV strain. Spectroscopic measurements of myocardial triglyceride content correlated significantly with histological analysis of biopsies obtained during aortic valve replacement. At 8.0 ± 2.1 months after aortic valve replacement, steatosis and strain had recovered toward normal. CONCLUSIONS: Pronounced myocardial steatosis is present in severe AS, regardless of symptoms, and is independently associated with the degree of LV strain impairment. Myocardial triglyceride content measured by magnetic resonance spectroscopy correlates with histological quantification. Steatosis and strain impairment are reversible after aortic valve replacement. Our findings suggest a novel pathophysiological mechanism in AS, myocardial steatosis, which may be amenable to treatment, thus potentially delaying onset of LV dysfunction.

Holloway CJ, Ntusi N, Suttie J, Mahmod M, Wainwright E, Clutton G, Hancock G, Beak P, Tajar A, Piechnik SK et al. 2013. Comprehensive cardiac magnetic resonance imaging and spectroscopy reveal a high burden of myocardial disease in HIV patients. Circulation, 128 (8), pp. 814-822. | Show Abstract | Read more

BACKGROUND: HIV infection continues to be endemic worldwide. Although treatments are successful, it remains controversial whether patients receiving optimal therapy have structural, functional, or biochemical cardiac abnormalities that may underlie their increased cardiac morbidity and mortality. The purpose of this study was to characterize myocardial abnormalities in a contemporary group of HIV-infected individuals undergoing combination antiretroviral therapy. METHODS AND RESULTS: Volunteers with HIV who were undergoing combination antiretroviral therapy and age-matched control subjects without a history of cardiovascular disease underwent cardiac magnetic resonance imaging and spectroscopy for the determination of cardiac function, myocardial fibrosis, and myocardial lipid content. A total of 129 participants were included in this analysis. Compared with age-matched control subjects (n=39; 30.23%), HIV-infected subjects undergoing combination antiretroviral therapy (n=90; 69.77%) had 47% higher median myocardial lipid levels (P <0.003) and 74% higher median plasma triglyceride levels (both P<0.001). Myocardial fibrosis, predominantly in the basal inferolateral wall of the left ventricle, was observed in 76% of HIV-infected subjects compared with 13% of control subjects (P<0.001). Peak myocardial systolic and diastolic longitudinal strain were also lower in HIV-infected individuals than in control subjects and remained statistically significant after adjustment for available confounders. CONCLUSIONS: Comprehensive cardiac imaging revealed cardiac steatosis, alterations in cardiac function, and a high prevalence of myocardial fibrosis in a contemporary group of asymptomatic HIV-infected subjects undergoing combination antiretroviral therapy. Cardiac steatosis and fibrosis may underlie cardiac dysfunction and increased cardiovascular morbidity and mortality in subjects with HIV.

Faller KM, Medway DJ, Aksentijevic D, Sebag-Montefiore L, Schneider JE, Lygate CA, Neubauer S. 2013. Ribose Supplementation Alone or with Elevated Creatine Does Not Preserve High Energy Nucleotides or Cardiac Function in the Failing Mouse Heart. PLoS One, 8 (6), pp. e66461. | Show Abstract | Read more

BACKGROUND: Reduced levels of creatine and total adenine nucleotides (sum of ATP, ADP and AMP) are hallmarks of chronic heart failure and restoring these pools is predicted to be beneficial by maintaining the diseased heart in a more favourable energy state. Ribose supplementation is thought to support both salvage and re-synthesis of adenine nucleotides by bypassing the rate-limiting step. We therefore tested whether ribose would be beneficial in chronic heart failure in control mice and in mice with elevated myocardial creatine due to overexpression of the creatine transporter (CrT-OE). METHODS AND RESULTS: FOUR GROUPS WERE STUDIED: sham; myocardial infarction (MI); MI+ribose; MI+CrT-OE+ribose. In a pilot study, ribose given in drinking water was bioavailable, resulting in a two-fold increase in myocardial ribose-5-phosphate levels. However, 8 weeks post-surgery, total adenine nucleotide (TAN) pool was decreased to a similar amount (8-14%) in all infarcted groups irrespective of the treatment received. All infarcted groups also presented with a similar and substantial degree of left ventricular (LV) dysfunction (3-fold reduction in ejection fraction) and LV hypertrophy (32-47% increased mass). Ejection fraction closely correlated with infarct size independently of treatment (r(2) = 0.63, p<0.0001), but did not correlate with myocardial creatine or TAN levels. CONCLUSION: Elevating myocardial ribose and creatine levels failed to maintain TAN pool or improve post-infarction LV remodeling and function. This suggests that ribose is not rate-limiting for purine nucleotide biosynthesis in the chronically failing mouse heart and that alternative strategies to preserve TAN pool should be investigated.

Carapella V, Bordas R, Pathmanathan P, Schneider JE, Kohl P, Burrage K, Grau V. 2013. Effect of Fibre Orientation Optimisation in an Electromechanical Model of Left Ventricular Contraction in Rat FUNCTIONAL IMAGING AND MODELING OF THE HEART, 7945 pp. 46-53. | Show Abstract | Read more

Subject-specific, or personalised, modelling is one of the main targets in current cardiac modelling research. The aim of this study is to assess the improvement in predictive power gained by introducing subject-specific fibre models within an electromechanical model of left ventricular contraction in rat. A quantitative comparison of a series of global rule-based fibre models with an image-based locally optimised fibre model was performed. Our results show small difference in the predicted values of ejection fraction, wall thickening and base-to-apex shortening between the fibre models considered. In comparison, much larger differences appear between predicted values and those measured in experimental images. Further study of the constitutive behaviour and architecture of cardiac tissue is required before electromechanical models can fully benefit from the introduction of subject-specific fibres. Additionally, our study shows that, in the current model, an orthotropic description of the tissue is preferable to a transversely isotropic one, for the metrics considered. © 2013 Springer-Verlag.

Espe EKS, Aronsen JM, Skrbic B, Skulberg VM, Schneider JE, Sejersted OM, Zhang L, Sjaastad I. 2013. Improved MR phase-contrast velocimetry using a novel nine-point balanced motion-encoding scheme with increased robustness to eddy current effects Magnetic Resonance in Medicine, 69 (1), pp. 48-61. | Show Abstract | Read more

Phase-contrast MRI (PC-MRI) velocimetry is a noninvasive, high-resolution motion assessment tool. However, high motion sensitivity requires strong motion-encoding magnetic gradients, making phase-contrast-MRI prone to baseline shift artifacts due to the generation of eddy currents. In this study, we propose a novel nine-point balanced velocity-encoding strategy, designed to be more accurate in the presence of strong and rapidly changing gradients. The proposed method was validated using a rotating phantom, and its robustness and precision were explored and compared with established approaches through computer simulations and in vivo experiments. Computer simulations yielded a 39-57% improvement in velocity-noise ratio (corresponding to a 27-33% reduction in measurement error), depending on which method was used for comparison. Moreover, in vivo experiments confirmed this by demonstrating a 26-53% reduction in accumulated velocity error over the R-R interval. The nine-point balanced phase-contrast-MRI-encoding strategy is likely useful for settings where high spatial and temporal resolution and/or high motion sensitivity is required, such as in high-resolution rodent myocardial tissue phase mapping. Magn Reson Med, 2013. © 2012 Wiley Periodicals, Inc. Copyright © 2012 Wiley Periodicals, Inc.

Burton RA, Schneider JE, Bishop MJ, Hales PW, Bollensdorff C, Robson MD, Wong KC, Morris J, Quinn TA, Kohl P. 2012. Microscopic magnetic resonance imaging reveals high prevalence of third coronary artery in human and rabbit heart. Europace, 14 Suppl 5 (suppl_5), pp. v73-v81. | Show Abstract | Read more

AIM: The human coronary tree is commonly assumed to have two roots: the left and right coronary arteries (LCA and RCA, respectively). However, a third coronary artery (TCA) has been observed in humans and animals, usually arising from the right anterior aortic sinus near the RCA. Using high-resolution magnetic resonance imaging, we identified TCA prevalence and characteristics in rabbit and human hearts. METHODS AND RESULTS: Third coronary artery presence was analysed in hearts from 11 New Zealand white rabbits and 7 human cadavers, using excised tissue that was fixed, gadolinium-treated, and agar-embedded for imaging-based reconstruction. A TCA was identified in all rabbit hearts and six of seven human hearts, originating either from an independent ostium (7 of 11 rabbits, 2 of 7 humans) or an ostium shared with the RCA (4 of 11 rabbits, 4 of 7 humans). Proximal TCA cross-sectional area in rabbits was 15.3 ± 6.0% of RCA area (mean ± SD, based on n = 9 rabbit hearts in which reliable measurements could be taken for both vessels), and 26.7 ± 10.1% in humans (n = 4). In all-but-one case where a TCA was observed, it originated ventral to the RCA, progressing towards the right ventricular outflow tract. In one rabbit, the TCA originated dorsal to the RCA and progressed towards the Crista terminalis in the right atrium. A fourth vessel, forming a separate aortic Vas vasorum was occasionally seen, originating from the right anterior aortic sinus either from an ostium common with (1 of 11 rabbits, 0 of 7 humans) or independent of (1 of 11 rabbits, 1 of 7 humans) the TCA. Pilot optical mapping experiments showed that TCA occlusion had variable acute effects on rabbit cardiac electrophysiology. CONCLUSION: Third coronary artery presence is common in rabbit and human hearts. Functional effects of disrupted TCA blood supply are ill-investigated, and the rabbit may be a suitable species for such research.

Lygate CA, Schneider JE, Neubauer S. 2013. Investigating cardiac energetics in heart failure. Exp Physiol, 98 (3), pp. 601-605. | Show Abstract | Read more

The energetic requirements of the heart are, weight for weight, higher than for any other organ. The heart provides non-stop function for a lifetime, while maintaining energy in reserve in order to respond to increased demand. This demand is met by continuously recycling a relatively small pool of ATP, with the creatine kinase (CK) system acting as a spatial and temporal buffer. In the failing heart, key components of this system are downregulated, but whether these energetic changes are biomarkers or drivers of dysfunction and whether they represent therapeutic targets are the subjects of ongoing research. Key methodologies are now becoming available in vivo to help address these questions in mouse models, such as (31)P magnetic resonance spectroscopy to detect high-energy phosphates and (1)H magnetic resonance spectroscopy to detect total creatine. This report briefly discusses the challenges involved in using these technologies, the application and pitfalls of murine surgical models of heart failure, and how this has contributed to our understanding of pathophysiology in recent years.

Faller KM, Lygate CA, Neubauer S, Schneider JE. 2013. (1)H-MR spectroscopy for analysis of cardiac lipid and creatine metabolism. Heart Fail Rev, 18 (5), pp. 657-668. | Show Abstract | Read more

Magnetic resonance spectroscopy (MRS) is the only non-invasive, non-radiation-based technique for investigating the metabolism of living tissue. MRS of protons (1H-MRS), which provides the highest sensitivity of all MR-visible nuclei, is a method capable of detecting and quantifying specific cardiac biomolecules, such as lipids and creatine in normal and diseased hearts in both animal models and humans. This can be used to study mechanisms of heart failure development in a longitudinal manner, for example, the potential contribution of myocardial lipid accumulation in the context of ageing and obesity. Similarly, quantifying creatine levels provides insight into the energy storage and buffering capacity in the heart. Creatine depletion is consistently observed in heart failure independent of aetiology, but its contribution to pathophysiology remains a matter of debate. These and other questions can in theory be answered with cardiac MRS, but fundamental technical challenges have limited its use. The metabolites studied with MRS are much lower concentration than water protons, requiring methods to suppress the dominant water signal and resulting in larger voxel sizes and longer scan times compared to MRI. However, recent technical advances in MR hardware and software have facilitated the application of 1H-MRS in humans and animal models of heart disease as detailed in this review.

Lygate CA, Medway DJ, Ostrowski PJ, Aksentijevic D, Sebag-Montefiore L, Hunyor I, Zervou S, Schneider JE, Neubauer S. 2012. Chronic creatine kinase deficiency eventually leads to congestive heart failure, but severity is dependent on genetic background, gender and age. Basic Res Cardiol, 107 (5), pp. 276. | Show Abstract | Read more

The creatine kinase (CK) energy transport and buffering system supports cardiac function at times of high demand and is impaired in the failing heart. Mice deficient in muscle- and mitochondrial-CK (M/Mt-CK(-/-)) have previously been described, but exhibit an unexpectedly mild phenotype of compensated left ventricular (LV) hypertrophy. We hypothesised that heart failure would develop with age and performed echocardiography and LV haemodynamics at 1 year. Since all previous studies have utilised mice with a mixed genetic background, we backcrossed for >10 generations on to C57BL/6, and repeated the in vivo investigations. Male M/Mt-CK(-/-) mice on the mixed genetic background developed congestive heart failure as evidenced by significantly elevated end-diastolic pressure, impaired contractility, LV dilatation, hypertrophy and pulmonary congestion. Female mice were less severely affected, only showing trends for these parameters. After backcrossing, M/Mt-CK(-/-) mice had LV dysfunction consisting of impaired isovolumetric pressure changes and reduced contractile reserve, but did not develop congestive heart failure. Body weight was lower in knockout mice as a consequence of reduced total body fat. LV weight was not significantly elevated in relation to other internal organs and gene expression of LVH markers was normal, suggesting an absence of hypertrophy. In conclusion, the consequences of CK deficiency are highly dependent on genetic modifiers, gender and age. However, the observation that a primary defect in CK can, under the right conditions, result in heart failure suggests that impaired CK activity in the failing heart could contribute to disease progression.

Lygate CA, Bohl S, ten Hove M, Faller KM, Ostrowski PJ, Zervou S, Medway DJ, Aksentijevic D, Sebag-Montefiore L, Wallis J et al. 2012. Moderate elevation of intracellular creatine by targeting the creatine transporter protects mice from acute myocardial infarction. Cardiovasc Res, 96 (3), pp. 466-475. | Show Abstract | Read more

AIMS: Increasing energy storage capacity by elevating creatine and phosphocreatine (PCr) levels to increase ATP availability is an attractive concept for protecting against ischaemia and heart failure. However, testing this hypothesis has not been possible since oral creatine supplementation is ineffectual at elevating myocardial creatine levels. We therefore used mice overexpressing creatine transporter in the heart (CrT-OE) to test for the first time whether elevated creatine is beneficial in clinically relevant disease models of heart failure and ischaemia/reperfusion (I/R) injury. METHODS AND RESULTS: CrT-OE mice were selected for left ventricular (LV) creatine 20-100% above wild-type values and subjected to acute and chronic coronary artery ligation. Increasing myocardial creatine up to 100% was not detrimental even in ageing CrT-OE. In chronic heart failure, creatine elevation was neither beneficial nor detrimental, with no effect on survival, LV remodelling or dysfunction. However, CrT-OE hearts were protected against I/R injury in vivo in a dose-dependent manner (average 27% less myocardial necrosis) and exhibited greatly improved functional recovery following ex vivo I/R (59% of baseline vs. 29%). Mechanisms contributing to ischaemic protection in CrT-OE hearts include elevated PCr and glycogen levels and improved energy reserve. Furthermore, creatine loading in HL-1 cells did not alter antioxidant defences, but delayed mitochondrial permeability transition pore opening in response to oxidative stress, suggesting an additional mechanism to prevent reperfusion injury. CONCLUSION: Elevation of myocardial creatine by 20-100% reduced myocardial stunning and I/R injury via pleiotropic mechanisms, suggesting CrT activation as a novel, potentially translatable target for cardiac protection from ischaemia.

Hales PW, Schneider JE, Burton RA, Wright BJ, Bollensdorff C, Kohl P. 2012. Histo-anatomical structure of the living isolated rat heart in two contraction states assessed by diffusion tensor MRI. Prog Biophys Mol Biol, 110 (2-3), pp. 319-330. | Show Abstract | Read more

Deformation and wall-thickening of ventricular myocardium are essential for cardiac pump function. However, insight into the histo-anatomical basis for cardiac tissue re-arrangement during contraction is limited. In this report, we describe dynamic changes in regionally prevailing cardiomyocyte (fibre) and myolaminar (sheet) orientations, using Diffusion Tensor Imaging (DTI) of ventricles in the same living heart in two different mechanical states. Hearts, isolated from Sprague-Dawley rats, were Langendorff-perfused and imaged, initially in their slack state during cardioplegic arrest, then during lithium-induced contracture. Regional fibre- and sheet-orientations were derived from DTI-data on a voxel-wise basis. Contraction was accompanied with a decrease in left-handed helical fibres (handedness relative to the baso-apical direction) in basal, equatorial, and apical sub-epicardium (by 14.0%, 17.3%, 15.8% respectively; p < 0.001), and an increase in right-handed helical fibres of the sub-endocardium (by 11.0%, 12.1% and 16.1%, respectively; p < 0.001). Two predominant sheet-populations were observed, with sheet-angles of either positive (β+) or negative (β-) polarity relative to a 'chamber-horizontal plane' (defined as normal to the left ventricular long-axis). In contracture, mean 'intersection'-angle (geometrically quantifiable intersection of sheet-angle projections) between β+ and β- sheet-populations increased from 86.2 ± 5.5° (slack) to 108.3 ± 5.4° (p < 0.001). Subsequent high-resolution DTI of fixed myocardium, and histological sectioning, reconfirmed the existence of alternating sheet-plane populations. Our results suggest that myocardial tissue layers in alternating sheet-populations align into a more chamber-horizontal orientation during contraction. This re-arrangement occurs via an accordion-like mechanism that, combined with inter-sheet slippage, can significantly contribute to ventricular deformation, including wall-thickening in a predominantly centripetal direction and baso-apical shortening.

McAteer MA, Mankia K, Ruparelia N, Jefferson A, Nugent HB, Stork LA, Channon KM, Schneider JE, Choudhury RP. 2012. A leukocyte-mimetic magnetic resonance imaging contrast agent homes rapidly to activated endothelium and tracks with atherosclerotic lesion macrophage content. Arterioscler Thromb Vasc Biol, 32 (6), pp. 1427-1435. | Show Abstract | Read more

OBJECTIVE: Endothelial cell activation is an important mediator of monocyte recruitment to sites of vascular inflammation. We hypothesized that high-affinity dual-ligand microparticles of iron oxide (MPIO), targeted to P-selectin and vascular cell adhesion molecule-1 (PV-MPIO), would identify activated endothelial cells during atherosclerosis progression. METHODS AND RESULTS: In vivo magnetic resonance imaging in apolipoprotein E-deficient mice showed rapid binding of PV-MPIO to the aortic root, which was maximal 30 minutes post-MPIO injection and maintained at 60 minutes. Minimal binding was observed for control IgG-MPIO. Intensely low magnetic resonance signal areas, corresponding to PV-MPIO binding, were detected early (14 weeks), during foam cell formation. Contrast effects increased at 20 weeks during fibrofatty lesion development (P<0.05), but reduced by 30 weeks (P<0.01). Across all lesion severities, magnetic resonance imaging contrast effects correlated with lesion macrophage area quantified by immunohistochemistry (R=0.53; P<0.01). Near-infrared fluorescently labeled PV-MPIO were shown, by flow cytometry, to bind only activated endothelial cells and not to macrophages. Using en face immunofluorescence, we further demonstrate selective PV-MPIO accumulation at atherosclerosis-susceptible sites, with minimal binding to atherosclerosis-spared regions. CONCLUSIONS: This high-affinity leukocyte-mimetic magnetic resonance imaging agent reveals endothelial activation. PV-MPIO demonstrate exceptionally rapid in vivo steady state accumulation, providing conspicuous magnetic resonance contrast effects that can be objectively quantified. In atherosclerosis progression, PV-MPIO tracked closely with the burden and distribution of plaque macrophages, not merely plaque size. On a biocompatible platform, this approach has potential for quantitative magnetic resonance imaging of inflammatory disease activity.

Jung B, Odening KE, Dall'Armellina E, Föll D, Menza M, Markl M, Schneider JE. 2012. A quantitative comparison of regional myocardial motion in mice, rabbits and humans using in-vivo phase contrast CMR. J Cardiovasc Magn Reson, 14 (1), pp. 87. | Show Abstract | Read more

BACKGROUND: Genetically manipulated animals like mice or rabbits play an important role in the exploration of human cardiovascular diseases. It is therefore important to identify animal models that closely mimic physiological and pathological human cardiac function. METHODS: In-vivo phase contrast cardiovascular magnetic resonance (CMR) was used to measure regional three-directional left ventricular myocardial motion with high temporal resolution in mice (N=18), rabbits (N=8), and humans (N=20). Radial, long-axis, and rotational myocardial velocities were acquired in left ventricular basal, mid-ventricular, and apical short-axis locations. RESULTS: Regional analysis revealed different patterns of motion: 1) In humans and rabbits, the apex showed slower radial velocities compared to the base. 2) Significant differences within species were seen in the pattern of long-axis motion. Long-axis velocities during systole were fairly homogeneously distributed in mice, whereas humans showed a dominant component in the lateral wall and rabbits in the base. 3) Rotational velocities and twist showed the most distinct patterns in both temporal evolution and relative contribution of base, mid-ventricle and apex, respectively. Interestingly, a marked difference in rotational behavior during early-systole was found in mice, which exhibited clockwise rotation in all slice locations compared to counter-clockwise rotation in rabbits and humans. CONCLUSIONS: Phase contrast CMR revealed subtle, but significantly different regional myocardial motion patterns in mice, rabbits and humans. This finding has to be considered when investigating myocardial motion pattern in small animal models of heart disease.

Gibb M, Burton RAB, Bollensdorff C, Afonso C, Mansoori T, Schotten U, Gavaghan DJ, Rodriguez B, Schneider JE, Kohl P, Grau V. 2012. Resolving the three-dimensional histology of the heart Lecture Notes in Computer Science (including subseries Lecture Notes in Artificial Intelligence and Lecture Notes in Bioinformatics), 7605 LNBI pp. 2-16. | Show Abstract | Read more

Cardiac histo-anatomical structure is a key determinant in all aspects of cardiac function. While some characteristics of micro- and macrostructure can be quantified using non-invasive imaging methods, histology is still the modality that provides the best combination of resolution and identification of cellular/sub-cellular substrate identities. The main limitation of histology is that it does not provide inherently consistent three-dimensional (3D) volume representations. This paper presents methods developed within our group to reconstruct 3D histological datasets. It includes the use of high-resolution MRI and block-face images to provide supporting volumetric datasets to guide spatial reintegration of 2D histological section data, and presents recent developments in sample preparation, data acquisition, and image processing. © 2012 Springer-Verlag.

Chen CM, Bentham J, Cosgrove C, Braganca J, Cuenda A, Bamforth SD, Schneider JE, Watkins H, Keavney B, Davies B, Bhattacharya S. 2012. Functional significance of SRJ domain mutations in CITED2. PLoS One, 7 (10), pp. e46256. | Show Abstract | Read more

CITED2 is a transcriptional co-activator with 3 conserved domains shared with other CITED family members and a unique Serine-Glycine Rich Junction (SRJ) that is highly conserved in placental mammals. Loss of Cited2 in mice results in cardiac and aortic arch malformations, adrenal agenesis, neural tube and placental defects, and partially penetrant defects in left-right patterning. By screening 1126 sporadic congenital heart disease (CHD) cases and 1227 controls, we identified 19 variants, including 5 unique non-synonymous sequence variations (N62S, R92G, T166N, G180-A187del and A187T) in patients. Many of the CHD-specific variants identified in this and previous studies cluster in the SRJ domain. Transient transfection experiments show that T166N mutation impairs TFAP2 co-activation function and ES cell proliferation. We find that CITED2 is phosphorylated by MAPK1 in vitro at T166, and that MAPK1 activation enhances the coactivation function of CITED2 but not of CITED2-T166N. In order to investigate the functional significance in vivo, we generated a T166N mutation of mouse Cited2. We also used PhiC31 integrase-mediated cassette exchange to generate a Cited2 knock-in allele replacing the mouse Cited2 coding sequence with human CITED2 and with a mutant form deleting the entire SRJ domain. Mouse embryos expressing only CITED2-T166N or CITED2-SRJ-deleted alleles surprisingly show no morphological abnormalities, and mice are viable and fertile. These results indicate that the SRJ domain is dispensable for these functions of CITED2 in mice and that mutations clustering in the SRJ region are unlikely to be the sole cause of the malformations observed in patients with sporadic CHD. Our results also suggest that coding sequence mutations observed in case-control studies need validation using in vivo models and that predictions based on structural conservation and in vitro functional assays, or even in vivo global loss of function models, may be insufficient.

Fortune S, Jansen MA, Anderson T, Gray GA, Schneider JE, Hoskins PR, Marshall I. 2012. Development and characterization of rodent cardiac phantoms: Comparison with in vivo cardiac imaging Magnetic Resonance Imaging, 30 (8), pp. 1186-1191. | Show Abstract | Read more

The increasing availability of rodent models of human cardiovascular disease has led to a need to translate noninvasive imaging techniques such as magnetic resonance imaging (MRI) from the clinic to the animal laboratory. The aim of this study was to develop phantoms simulating the short-axis view of left ventricular motion of rats and mice, thus reducing the need for live animals in the development of MRI. Cylindrical phantoms were moulded from polyvinyl alcohol (PVA) Cryogel and attached via stiff water-filled tubing to a gear pump. Pulsatile distension of the phantoms was effected by suitable programming of the pump. Cine MRI scanning was carried out at 7 T and compared with in vivo rodent cardiac imaging. Suitable pulsatile performance was achieved with phantoms for which the PVA material had been subjected to two freeze-thaw cycles, resulting in T1 and T2 relaxation time constants of 1656±124 ms and 55±10 ms, respectively. For the rat phantom operating at 240 beats per min (bpm), the dynamic range of the outer diameter was from 10.3 to 12.4 mm with the wall thickness varying between 1.9 and 1.2 mm. Corresponding figures for the mouse phantom at 480 bpm were outer diameter range from 5.4 to 6.4 mm and wall thickness from 1.5 to 1.2 mm. Dynamic cardiac phantoms s imulating rodent left ventricular motion in the short-axis view were successfully developed and compared with in vivo imaging. The phantoms can be used for future development work with reduced need of live animals. © 2012 Elsevier Inc.

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McAteer MA, Mankia K, Ruparelia N, Jefferson A, Nugent HB, Stork LA, Channon KM, Schneider JE, Choudhury RP. 2012. A leukocyte-mimetic magnetic resonance imaging contrast agent homes rapidly to activated endothelium and tracks with atherosclerotic lesion macrophage content Arteriosclerosis, Thrombosis, and Vascular Biology, 32 (6), pp. 1427-1435. | Show Abstract | Read more

Objective-Endothelial cell activation is an important mediator of monocyte recruitment to sites of vascular inflammation. We hypothesized that high-affinity dual-ligand microparticles of iron oxide (MPIO), targeted to P-selectin and vascular cell adhesion molecule-1 (PV-MPIO), would identify activated endothelial cells during atherosclerosis progression. Methods and Results-In vivo magnetic resonance imaging in apolipoprotein E-deficient mice showed rapid binding of PV-MPIO to the aortic root, which was maximal 30 minutes post-MPIO injection and maintained at 60 minutes. Minimal binding was observed for control IgG-MPIO. Intensely low magnetic resonance signal areas, corresponding to PV-MPIO binding, were detected early (14 weeks), during foam cell formation. Contrast effects increased at 20 weeks during fibrofatty lesion development (P < 0.05), but reduced by 30 weeks (P < 0.01). Across all lesion severities, magnetic resonance imaging contrast effects correlated with lesion macrophage area quantified by immunohistochemistry (R=0.53; P < 0.01). Near-infrared fluorescently labeled PV-MPIO were shown, by flow cytometry, to bind only activated endothelial cells and not to macrophages. Using en face immunofluorescence, we further demonstrate selective PV-MPIO accumulation at atherosclerosis-susceptible sites, with minimal binding to atherosclerosis- spared regions. Conclusion-This high-affinity leukocyte-mimetic magnetic resonance imaging agent reveals endothelial activation. PV-MPIO demonstrate exceptionally rapid in vivo steady state accumulation, providing conspicuous magnetic resonance contrast effects that can be objectively quantified. In atherosclerosis progression, PV-MPIO tracked closely with the burden and distribution of plaque macrophages, not merely plaque size. On a biocompatible platform, this approach has potential for quantitative magnetic resonance imaging of inflammatory disease activity. © 2012 American Heart Association, Inc.

MacDonald ST, Bamforth SD, Bragança J, Chen CM, Broadbent C, Schneider JE, Schwartz RJ, Bhattacharya S. 2013. A cell-autonomous role of Cited2 in controlling myocardial and coronary vascular development. Eur Heart J, 34 (32), pp. 2557-2565. | Show Abstract | Read more

AIMS: Myocardial development is dependent on concomitant growth of cardiomyocytes and a supporting vascular network. The coupling of myocardial and coronary vascular development is partly mediated by vascular endothelial growth factor (VEGFA) signalling and additional unknown mechanisms. We examined the cardiomyocyte specific role of the transcriptional co-activator Cited2 on myocardial microstructure and vessel growth, in relation to Vegfa expression. METHODS AND RESULTS: A cardiomyocyte-specific knockout of mouse Cited2 (Cited2(Nkx)) was analysed using magnetic resonance imaging and histology. Ventricular septal defects and significant compact layer thinning (P < 0.02 at right ventricular apex, P < 0.009 at the left ventricular apex in Cited2(Nkx) vs. controls, n = 11 vs. n = 7, respectively) were found. This was associated with a significant decrease in the number of capillaries to larger vessels (ratio 1.56 ± 0.56 vs. 3.25 ± 1.63, P = 2.7 × 10(-6) Cited2(Nkx) vs. controls, n = 11 vs. n = 7, respectively) concomitant with a 1.5-fold reduction in Vegfa expression (P < 0.02, Cited2(Nkx) vs. controls, n = 12 vs. n = 12, respectively). CITED2 was subsequently found at the Vegfa promoter in mouse embryonic hearts using chromatin immunoprecipitation, and moreover found to stimulate human VEGFA promoter activity in cooperation with TFAP2 transcription factors in transient transfection assays. There was no change in the myocardial expression of the left-right patterning gene Pitx2c, a previously known target of CITED2. CONCLUSIONS: This study delineates a novel cell-autonomous role of Cited2 in regulating VEGFA transcription and the development of myocardium and coronary vasculature in the mouse. We suggest that coupling of myocardial and coronary growth in the developing heart may occur in part through a Cited2→Vegfa pathway.

Dall'Armellina E, Jung BA, Lygate CA, Neubauer S, Markl M, Schneider JE. 2012. Improved method for quantification of regional cardiac function in mice using phase-contrast MRI Magnetic Resonance in Medicine, 67 (2), pp. 541-551. | Show Abstract | Read more

Phase-contrast magnetic resonance imaging is a technique that allows for characterization of regional cardiac function and for measuring transmural myocardial velocities in human hearts with high temporal and spatial resolution. The application of this technique (also known as tissue phase mapping) to murine hearts has been very limited so far. The aim of our study was to implement and to optimize tissue phase mapping for a comprehensive assessment of murine transmural wall motion. Baseline values for regional motion patterns in mouse hearts, based on the clinically used American Heart Association's 17-segment model, were established, and a detailed motion analysis of mouse heart for the entire cardiac cycle (including epicardial and endocardial motion patterns) is provided. Black-blood contrast was found to be essential to obtain reproducible velocity encoding. Tissue phase mapping of the mouse heart permits the detailed assessment of regional myocardial velocities. While a proof-of-principle application in a murine ischemia-reperfusion model was performed, future studies are warranted to assess its potential for the investigation of systolic and diastolic functions in genetically and surgically manipulated mouse models of human heart disease. Magn Reson Med, 2012. © 2011 Wiley Periodicals, Inc.

Wech T, Lemke A, Medway D, Stork LA, Lygate CA, Neubauer S, Köstler H, Schneider JE. 2011. Accelerating cine-MR imaging in mouse hearts using compressed sensing. J Magn Reson Imaging, 34 (5), pp. 1072-1079. | Show Abstract | Read more

PURPOSE: To combine global cardiac function imaging with compressed sensing (CS) in order to reduce scan time and to validate this technique in normal mouse hearts and in a murine model of chronic myocardial infarction. MATERIALS AND METHODS: To determine the maximally achievable acceleration factor, fully acquired cine data, obtained in sham and chronically infarcted (MI) mouse hearts were 2-4-fold undersampled retrospectively, followed by CS reconstruction and blinded image segmentation. Subsequently, dedicated CS sampling schemes were implemented at a preclinical 9.4 T magnetic resonance imaging (MRI) system, and 2- and 3-fold undersampled cine data were acquired in normal mouse hearts with high temporal and spatial resolution. RESULTS: The retrospective analysis demonstrated that an undersampling factor of three is feasible without impairing accuracy of cardiac functional parameters. Dedicated CS sampling schemes applied prospectively to normal mouse hearts yielded comparable left-ventricular functional parameters, and intra- and interobserver variability between fully and 3-fold undersampled data. CONCLUSION: This study introduces and validates an alternative means to speed up experimental cine-MRI without the need for expensive hardware.

Young AA, Medway DJ, Lygate CA, Neubauer S, Schneider JE. 2011. Accelerating global left-ventricular function assessment in mice using reduced slice acquisition and three-dimensional guide-point modelling. J Cardiovasc Magn Reson, 13 (1), pp. 49. | Show Abstract | Read more

BACKGROUND: To investigate the utility of three-dimensional guide-point modeling (GPM) to reduce the time required for CMR evaluation of global cardiac function in mice, by reducing the number of image slices required for accurate quantification of left-ventricular (LV) mass and volumes. METHODS: Five female C57Bl/6 mice 8 weeks post myocardial infarction induced by permanent occlusion of the left coronary artery, and six male control (un-operated) C57Bl/6 mice, were subject to CMR examination under isoflurane anaesthesia. Contiguous short axis (SAX) slices (1 mm thick 7-9 slices) were obtained together with two long axis (LAX) slices in two chamber and four chamber orientations. Using a mathematical model of the heart to interpolate information between the available slices, GPM LV mass and volumes were determined using full slice (all SAX and two LAX), six slice (four SAX and two LAX) and four slice (two SAX and two LAX) analysis protocols. All results were compared with standard manual volumetric analysis using all SAX slices. RESULTS: Infarct size was 39.1±5.1% of LV myocardium. No significant differences were found in left ventricular mass and volumes between the standard and GPM full and six slice protocols in infarcted mice (113±10, 116±11, and 117±11 mg respectively for mass), or between the standard and GPM full, six and four slice protocols in control mice, (105±14, 106±10, 104±12, and 105±7 mg respectively for mass). Significant differences were found in LV mass (135±18 mg) and EF using the GPM four slice protocol in infarcted mice (p<0.05). CONCLUSION: GPM enables accurate analysis of LV function in mice with relatively large infarcts using a reduced six slice acquisition protocol, and in mice with normal/symmetrical left-ventricular topology using a four slice protocol.

Hales PW, Burton RA, Bollensdorff C, Mason F, Bishop M, Gavaghan D, Kohl P, Schneider JE. 2011. Progressive changes in T₁, T₂ and left-ventricular histo-architecture in the fixed and embedded rat heart. NMR Biomed, 24 (7), pp. 836-843. | Show Abstract | Read more

Chemical tissue fixation, followed by embedding in either agarose or Fomblin, is common practice in time-intensive MRI studies of ex vivo biological samples, and is required to prevent tissue autolysis and sample motion. However, the combined effect of fixation and sample embedding may alter tissue structure and MRI properties. We investigated the progressive changes in T(1) and T(2) relaxation times, and the arrangement of locally prevailing cardiomyocyte orientation determined using diffusion tensor imaging, in embedded ex vivo rat hearts fixed using Karnovsky's solution (glutaraldehyde-formaldehyde mix). Three embedding media were investigated: (i) standard agarose (n = 3 hearts); (ii) Fomblin (n = 4 hearts); and (iii) iso-osmotic agarose (n = 3 hearts); in the latter, the osmolarity of the fixative and embedding medium was adjusted to 300 mOsm to match more closely that of native tissue. The T(1) relaxation time in the myocardium showed a pronounced decrease over a 48-h period following embedding in Fomblin (-11.3 ± 6.2%; mean ± standard deviation), but was stable in standard agarose- and iso-osmotic agarose-embedded hearts. The mean myocardial T(2) relaxation time increased in all embedded hearts: by 35.1 ± 14.7% with standard agarose embedding, 13.1 ± 5.6% with Fomblin and 13.3 ± 1.4% with iso-osmotic agarose. Deviation in the orientation of the primary eigenvector of the diffusion tensor occurred in all hearts (mean angular changes of 6.6°, 3.2° and 1.9° per voxel after 48 h in agarose-, Fomblin- and iso-osmotic agarose-embedded hearts, respectively), indicative of progressive structural changes in myocardial histo-architecture, in spite of previous exposure to fast-acting tissue fixation. Our results suggest that progressive structural changes occur in chemically fixed myocardium, and that the extent of these changes is modulated by the embedding medium, and by osmotic gradients between the fixative in the tissue and the surrounding medium.

Rial B, Robson MD, Neubauer S, Schneider JE. 2011. Rapid quantification of myocardial lipid content in humans using single breath-hold 1H MRS at 3 Tesla. Magn Reson Med, 66 (3), pp. 619-624. | Show Abstract | Read more

A rapid, proton magnetic resonance spectroscopy method to evaluate human myocardial lipid levels in a single breath-hold at 3 T using a commercial whole-body system is presented. During a 10 s breath-hold, water unsuppressed and suppressed spectra were acquired by two phased array coils using a short-echo time spectroscopic stimulated echo (STEAM) sequence electrocardiogram-triggered to mid-diastole. Lipid-to-water ratios were obtained in the septum of 15 healthy volunteers, (0.46 ± 0.19)%. These results agreed well with ratios obtained from averaged spectra acquired in seven multiple breath-holds, (0.45 ± 0.20)%, providing increased signal-to-noise ratio but requiring longer acquisition times. Excellent correlation was found between the two methods (r=0.94, P<0.05). Reproducibility of (1)H MRS for measuring myocardial lipid levels in a short breath-hold was acceptable in five repeated measurements within the same subject (coefficient of variation=19%). Thus, single breath-hold proton spectroscopy allows reliable and quick quantification of myocardial lipids at 3 T.

Baghdadi L, Zamyadi M, Sled JG, Schneider JE, Bhattacharya S, Henkelman RM, Lerch JP. 2011. Semi-automatic segmentation of multiple mouse embryos in MR images. BMC Bioinformatics, 12 (1), pp. 237. | Show Abstract | Read more

BACKGROUND: The motivation behind this paper is to aid the automatic phenotyping of mouse embryos, wherein multiple embryos embedded within a single tube were scanned using Magnetic Resonance Imaging (MRI). RESULTS: Our algorithm, a modified version of the simplex deformable model of Delingette, addresses various issues with deformable models including initialization and inability to adapt to boundary concavities. In addition, it proposes a novel technique for automatic collision detection of multiple objects which are being segmented simultaneously, hence avoiding major leaks into adjacent neighbouring structures. We address the initialization problem by introducing balloon forces which expand the initial spherical models close to the true boundaries of the embryos. This results in models which are less sensitive to initial minimum of two fold after each stage of deformation. To determine collision during segmentation, our unique collision detection algorithm finds the intersection between binary masks created from the deformed models after every few iterations of the deformation and modifies the segmentation parameters accordingly hence avoiding collision.We have segmented six tubes of three dimensional MR images of multiple mouse embryos using our modified deformable model algorithm. We have then validated the results of the our semi-automatic segmentation versus manual segmentation of the same embryos. Our Validation shows that except paws and tails we have been able to segment the mouse embryos with minor error. CONCLUSIONS: This paper describes our novel multiple object segmentation technique with collision detection using a modified deformable model algorithm. Further, it presents the results of segmenting magnetic resonance images of up to 32 mouse embryos stacked in one gel filled test tube and creating 32 individual masks.

Dall'Armellina E, Jung BA, Lygate CA, Neubauer S, Markl M, Schneider JE. 2012. Improved method for quantification of regional cardiac function in mice using phase-contrast MRI. Magn Reson Med, 67 (2), pp. 541-551. | Show Abstract | Read more

Phase-contrast magnetic resonance imaging is a technique that allows for characterization of regional cardiac function and for measuring transmural myocardial velocities in human hearts with high temporal and spatial resolution. The application of this technique (also known as tissue phase mapping) to murine hearts has been very limited so far. The aim of our study was to implement and to optimize tissue phase mapping for a comprehensive assessment of murine transmural wall motion. Baseline values for regional motion patterns in mouse hearts, based on the clinically used American Heart Association's 17-segment model, were established, and a detailed motion analysis of mouse heart for the entire cardiac cycle (including epicardial and endocardial motion patterns) is provided. Black-blood contrast was found to be essential to obtain reproducible velocity encoding. Tissue phase mapping of the mouse heart permits the detailed assessment of regional myocardial velocities. While a proof-of-principle application in a murine ischemia-reperfusion model was performed, future studies are warranted to assess its potential for the investigation of systolic and diastolic functions in genetically and surgically manipulated mouse models of human heart disease.

Schneider JE. 2011. Assessment of global cardiac function. Methods Mol Biol, 771 pp. 387-405. | Show Abstract | Read more

High-resolution magnetic resonance cine imaging (cine-MRI) allows for a non-invasive assessment of ventricular function and mass in normal mice and in genetically and surgically modified mouse models of cardiac disease. The assessment of myocardial mass and function by cine-MRI does not rely on geometric assumptions, as the hearts are covered from the base to the apex, typically by a stack of two-dimensional images. The MR data acquisition is then followed by image segmentation of specific cine frames in each slice to obtain geometric and functional parameters, such as end-diastolic volume (EDV), end-systolic volume (ESV) or ejection fraction (EF). This technique has been well established in clinical routine application and it is now also becoming the reference method in experimental cardiovascular MRI. The cine images are typically acquired in short- and long-axis orientations of the heart to facilitate an accurate assessment of cardiac functional parameters. These views can be difficult to identify, particularly in animals with diseased hearts. Furthermore, data analysis can be the source of a systematic error, mainly for myocardial mass measurement. We have established protocols that allow for a quick and reproducible way of obtaining the relevant cardiac views for cine-MRI, and for accurate image analysis.

Karadag IE, Bishop M, Hales PW, Schneider JE, Kohl P, Gavaghan D, Grau V. 2011. Regionally Optimised Mathematical Models of Cardiac Myocyte Orientation in Rat Hearts FUNCTIONAL IMAGING AND MODELING OF THE HEART, 6666 pp. 294-301. | Show Abstract | Read more

Mathematical models of ventricular cardiomyocyte orientation provide a simple description of histo-anatomical arrangements that are important for cardiac mechano-electric behaviour. They can be used to analyse interspecies differences, to explore dynamic remodelling such as during development or disease, and they are key for building realistic computational representations of the heart. This study investigates the suitability of regionally optimised models to represent accurately myocardial structure. Using DT-MRI scans as a reference, we calculate an optimised model by finding the parameters that minimise angular differences, both globally and regionally using a 16-segment topography. Results show angular differences between the optimized model and DT-MRI data of up to 15 degrees, with regional optimization providing a clear improvement in model accuracy (up to 52% error reduction). © 2011 Springer-Verlag Berlin Heidelberg.

Aksentijević D, Lygate CA, Makinen K, Zervou S, Sebag-Montefiore L, Medway D, Barnes H, Schneider JE, Neubauer S. 2010. High-energy phosphotransfer in the failing mouse heart: role of adenylate kinase and glycolytic enzymes. Eur J Heart Fail, 12 (12), pp. 1282-1289. | Show Abstract | Read more

AIMS: To measure the activity of the key phosphotransfer enzymes creatine kinase (CK), adenylate kinase (AK), and glycolytic enzymes in two common mouse models of chronic heart failure. METHODS AND RESULTS: C57BL/6 mice were subjected to transverse aortic constriction (TAC), myocardial infarction induced by coronary artery ligation (CAL), or sham operation. Activities of phosphotransfer enzymes CK, AK, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 3-phosphoglycerate kinase (PGK), and pyruvate kinase were assessed spectrophotometrically. Mice were characterized by echocardiography or magnetic resonance imaging 5- to 8-week post-surgery and selected for the presence of congestive heart failure. All mice had severe left ventricular hypertrophy, impaired systolic function and pulmonary congestion compared with sham controls. A significant decrease in myocardial CK and maximal CK reaction velocity was observed in both experimental models of heart failure. However, the activity of AK and its isoforms remained unchanged, despite a reduction in its protein expression. In contrast, the activities of glycolytic phosphotransfer mediators GAPDH and PGK were 19 and 12% higher in TAC, and 31 and 23% higher in CAL models, respectively. CONCLUSION: Chronic heart failure in the mouse is characterized by impaired CK function, unaltered AK, and increased activity of glycolytic phosphotransfer enzymes. This pattern of altered phosphotransfer activity was observed independent of the heart failure aetiology.

Asopa S, Cagampang FR, Anthony FW, Lanham SA, Schneider JE, Ohri SK, Hanson MA. 2010. Effect of a low-protein diet during pregnancy on expression of genes involved in cardiac hypertrophy in fetal and adult mouse offspring. J Dev Orig Health Dis, 1 (6), pp. 371-375. | Show Abstract | Read more

Gene markers for cardiomyocyte growth, proliferation and remodeling were examined in mouse fetuses and adult male offspring exposed to maternal low-protein (LP) diet during pregnancy. Whole heart volume, measured by magnetic resonance imaging, was smaller in day 15 LP fetuses v. those from chow-fed dams (C), whereas heart volume was greater in adult LP v. C offspring. These LP offspring were hypertensive and had larger cardiomyocytes v. C animals. The mRNA levels of cyclin G1, a marker for cell growth, were lower in LP fetal hearts v. C hearts, but similar in the left ventricle of adult LP and C offspring. Opposite trends were found in brain natriuretic peptide levels (a marker of cardiac hypertrophy). Thus, maternal LP during pregnancy results in smaller fetal hearts and is accompanied by changes in expression of genes involved in cardiomyocyte growth, which are associated with cardiac hypertrophy and hypertension in adulthood.

McAteer MA, Mankia K, Hagen R, Stork L-AA, Schneider JE, Choudhury RP. 2010. Noninvasive Molecular Imaging of Differential Endothelial Adhesion Molecule Expression in Early Atherosclerosis Correlates With Plaque Macrophage Content in Apo E Knockout Mice CIRCULATION, 122 (21),

Zamyadi M, Baghdadi L, Lerch JP, Bhattacharya S, Schneider JE, Henkelman RM, Sled JG. 2010. Mouse embryonic phenotyping by morphometric analysis of MR images. Physiol Genomics, 42A (2), pp. 89-95. | Show Abstract | Read more

A new method is described for automatic detection of subtle morphological phenotypes in mouse embryos. Based on high-resolution magnetic resonance imaging scanning and nonlinear image alignment, this method is demonstrated by comparing the morphology of two inbred strains, C57BL/6J and 129Sv/S1ImJ, at 15.5 days postconception. Mouse embryo morphology was found to be highly amenable to this kind of analysis with very low levels (on average 110 μm) of residual anatomical variation within strains after linear differences in pose and scale are removed. Mapping of local size differences showed that C57BL/6J embryos were larger than 129Sv/S1ImJ embryos, although these differences were not uniformly distributed across the anatomy. Expressed in terms of organ volumes, heart and lung were larger in C57BL/6J embryos, while brain and liver were comparable in volume between strains. The positive relationship between organ size and embryo size was consistent for the two strains but differed by organ, with the brain and liver being the least variable. Together these findings suggest the power of this technique for detecting subtle phenotypic differences arising from mutated genes.

Bentham J, Michell AC, Lockstone H, Andrew D, Schneider JE, Brown NA, Bhattacharya S. 2010. Maternal high-fat diet interacts with embryonic Cited2 genotype to reduce Pitx2c expression and enhance penetrance of left-right patterning defects. Hum Mol Genet, 19 (17), pp. 3394-3401. | Show Abstract | Read more

Deficiency of the transcription factor Cited2 in mice results in cardiac malformation, adrenal agenesis, neural tube, placental defects and partially penetrant cardiopulmonary laterality defects resulting from an abnormal Nodal->Pitx2c pathway. Here we show that a maternal high-fat diet more than doubles the penetrance of laterality defects and, surprisingly, induces palatal clefting in Cited2-deficient embryos. Both maternal diet and Cited2 deletion reduce embryo weight and kidney and thymus volume. Expression profiling identified 40 embryonic transcripts including Pitx2 that were significantly affected by embryonic genotype-maternal diet interaction. We show that a high-fat diet reduces Pitx2c levels >2-fold in Cited2-deficient embryos. Taken together, these results define a novel interaction between maternal high-fat diet and embryonic Cited2 deficiency that affects Pitx2c expression and results in abnormal laterality. They suggest that appropriate modifications of maternal diet may prevent such defects in humans.

Schneider JE, Lanz T, Barnes H, Stork LA, Bohl S, Lygate CA, Ordidge RJ, Neubauer S. 2011. Accelerated cardiac magnetic resonance imaging in the mouse using an eight-channel array at 9.4 Tesla. Magn Reson Med, 65 (1), pp. 60-70. | Show Abstract | Read more

MRI has become an important tool to noninvasively assess global and regional cardiac function, infarct size, or myocardial blood flow in surgically or genetically modified mouse models of human heart disease. Constraints on scan time due to sensitivity to general anesthesia in hemodynamically compromised mice frequently limit the number of parameters available in one imaging session. Parallel imaging techniques to reduce acquisition times require coil arrays, which are technically challenging to design at ultrahigh magnetic field strengths. This work validates the use of an eight-channel volume phased-array coil for cardiac MRI in mice at 9.4 T. Two- and three-dimensional sequences were combined with parallel imaging techniques and used to quantify global cardiac function, T(1)-relaxation times and infarct sizes. Furthermore, the rapid acquisition of functional cine-data allowed for the first time in mice measurement of left-ventricular peak filling and ejection rates under intravenous infusion of dobutamine. The results demonstrate that a threefold accelerated data acquisition is generally feasible without compromising the accuracy of the results. This strategy may eventually pave the way for routine, multiparametric phenotyping of mouse hearts in vivo within one imaging session of tolerable duration.

Michell AC, Bragança J, Broadbent C, Joyce B, Franklyn A, Schneider JE, Bhattacharya S, Bamforth SD. 2010. A novel role for transcription factor Lmo4 in thymus development through genetic interaction with Cited2. Dev Dyn, 239 (7), pp. 1988-1994. | Show Abstract | Read more

Deletion of the transcriptional modulator Cited2 in the mouse results in embryonic lethality, cardiovascular malformations, adrenal agenesis, cranial ganglia fusion, exencephaly, and left-right patterning defects, all seen with a varying degree of penetrance. The phenotypic heterogeneity, observed on different genetic backgrounds, indicates the existence of both genetic and environmental modifiers. Mice lacking the LIM domain-containing protein Lmo4 share specific phenotypes with Cited2 null embryos, such as embryonic lethality, cranial ganglia fusion, and exencephaly. These shared phenotypes suggested that Lmo4 may be a potential genetic modifier of the Cited2 phenotype. Examination of Lmo4-deficient embryos revealed partially penetrant cardiovascular malformations and hypoplastic thymus. Examination of Lmo4;Cited2 compound mutants indicated that there is a genetic interaction between Cited2 and Lmo4 in control of thymus development. Our data suggest that this may occur, in part, through control of expression of a common target gene, Tbx1, which is necessary for normal thymus development.

Lanz T, Müller M, Barnes H, Neubauer S, Schneider JE. 2010. A high-throughput eight-channel probe head for murine MRI at 9.4 T. Magn Reson Med, 64 (1), pp. 80-87. | Show Abstract | Read more

Murine MRI studies are conducted on dedicated MR systems, typically equipped with ultra-high-field magnets (>or=4.7 T; bore size: approximately 12-25 cm), using a single transmit-receive coil (volume or surface coil in linear or quadrature mode) or a transmit-receive coil combination. Here, we report on the design and characterization of an eight-channel volume receive-coil array for murine MRI at 400 MHz. The array was combined with a volume-transmit coil and integrated into one probe head. Therefore, the animal handling is fully decoupled from the radiofrequency setup. Furthermore, fixed tune and match of the coils and a reduced number of connectors minimized the setup time. Optimized preamplifier design was essential for minimizing the noise coupling between the elements. A comprehensive characterization of transmit volume resonator and receive coil array is provided. The performance of the coil array is compared to a quadrature-driven birdcage coil with identical sensitive volume. It is shown that the miniature size of the elements resulted in coil noise domination and therefore reduced signal-to-noise-ratio performance in the center compared to the quadrature birdcage. However, it allowed for 3-fold accelerated imaging of mice in vivo, reducing scan time requirements and thus increasing the number of mice that can be scanned per unit of time.

Zhang X, Schneider JE, Portnoy S, Bhattacharya S, Henkelman RM. 2010. Comparative SNR for high-throughput mouse embryo MR microscopy. Magn Reson Med, 63 (6), pp. 1703-1707. | Show Abstract | Read more

MR microscopy is being explored as a useful imaging tool to phenotype mouse embryos due to its volume coverage with three-dimensional isotropic resolution. However, the main limitation for mouse embryo MR microscopy is the signal-to-noise ratio. Large numbers of embryos are needed for phenotypic screening, making high throughput essential. Two high-throughput imaging approaches, multi-embryo shared-coil (shared) and multi-embryo individual-coil (individual), have been developed for phenotyping mouse embryos. This study quantitatively compares the signal-to-noise ratio at equivalent times between these two established methods by compensating for differences that result from field strength. While the individual method provides 3.3 times as much signal-to-noise ratio as the shared method at equivalent conditions, it is more difficult and expensive to implement. Furthermore, the number of embryos that can be imaged concurrently is limited by the number of receiver channels. The objective of this study is to provide measured comparative data to guide choices for high-throughput mouse embryo MR microscopy and other similar applications.

Phillips D, Ten Hove M, Schneider JE, Wu CO, Sebag-Montefiore L, Aponte AM, Lygate CA, Wallis J, Clarke K, Watkins H et al. 2010. Mice over-expressing the myocardial creatine transporter develop progressive heart failure and show decreased glycolytic capacity. J Mol Cell Cardiol, 48 (4), pp. 582-590. | Show Abstract | Read more

The metabolic phenotype of the failing heart includes a decrease in phosphocreatine and total creatine concentration [Cr], potentially contributing to contractile dysfunction. Surprisingly, in 32- week-old mice over-expressing the myocardial creatine transporter (CrT-OE), we previously demonstrated that elevated [Cr] correlates with left ventricular (LV) hypertrophy and failure. The aim of this study was to determine the temporal relationship between elevated [Cr] and the onset of cardiac dysfunction and to screen for potential molecular mechanisms. CrT-OE mice were compared with wild-type (WT) littermate controls longitudinally using cine-MRI to measure cardiac function and single-voxel (1)H-MRS to measure [Cr] in vivo at 6, 16, 32, and 52 weeks of age. CrT-OE mice had elevated [Cr] at 6 weeks (mean 1.9-fold), which remained constant throughout life. Despite this increased [Cr], LV dysfunction was not apparent until 16 weeks and became more pronounced with age. Additionally, LV tissue from 12 to 14 week old CrT-OE mice was compared to WT using 2D difference in-gel electrophoresis (DIGE). These analyses detected a majority of the heart's metabolic enzymes and identified seven proteins that were differentially expressed between groups. The most pronounced protein changes were related to energy metabolism: alpha- and beta-enolase were selectively decreased (p<0.05), while the remaining enzymes of glycolysis were unchanged. Consistent with a decrease in enolase content, its activity was significantly lower in CrT-OE hearts (in WT, 0.59+/-0.02 micromol ATP produced/microg protein/min; CrT-OE, 0.31+/-0.06; p<0.01). Additionally, anaerobic lactate production was decreased in CrT-OE mice (in WT, 102+/-3 micromol/g wet myocardium; CrT-OE, 78+/-13; p=0.02), consistent with decreased glycolytic capacity. Finally, we found that enolase may be regulated by increased expression of the beta-enolase repressor transcription factor, which was significantly increased in CrT-OE hearts. This study demonstrates that chronically increased myocardial [Cr] in the CrT-OE model leads to the development of progressive hypertrophy and heart failure, which may be mediated by a compromise in glycolytic capacity at the level of enolase.

Bishop MJ, Plank G, Burton RA, Schneider JE, Gavaghan DJ, Grau V, Kohl P. 2010. Development of an anatomically detailed MRI-derived rabbit ventricular model and assessment of its impact on simulations of electrophysiological function. Am J Physiol Heart Circ Physiol, 298 (2), pp. H699-H718. | Show Abstract | Read more

Recent advances in magnetic resonance (MR) imaging technology have unveiled a wealth of information regarding cardiac histoanatomical complexity. However, methods to faithfully translate this level of fine-scale structural detail into computational whole ventricular models are still in their infancy, and, thus, the relevance of this additional complexity for simulations of cardiac function has yet to be elucidated. Here, we describe the development of a highly detailed finite-element computational model (resolution: approximately 125 microm) of rabbit ventricles constructed from high-resolution MR data (raw data resolution: 43 x 43 x 36 microm), including the processes of segmentation (using a combination of level-set approaches), identification of relevant anatomical features, mesh generation, and myocyte orientation representation (using a rule-based approach). Full access is provided to the completed model and MR data. Simulation results were compared with those from a simplified model built from the same images but excluding finer anatomical features (vessels/endocardial structures). Initial simulations showed that the presence of trabeculations can provide shortcut paths for excitation, causing regional differences in activation after pacing between models. Endocardial structures gave rise to small-scale virtual electrodes upon the application of external field stimulation, which appeared to protect parts of the endocardium in the complex model from strong polarizations, whereas intramural virtual electrodes caused by blood vessels and extracellular cleft spaces appeared to reduce polarization of the epicardium. Postshock, these differences resulted in the genesis of new excitation wavefronts that were not observed in more simplified models. Furthermore, global differences in the stimulus recovery rates of apex/base regions were observed, causing differences in the ensuing arrhythmogenic episodes. In conclusion, structurally simplified models are well suited for a large range of cardiac modeling applications. However, important differences are seen when behavior at microscales is relevant, particularly when examining the effects of external electrical stimulation on tissue electrophysiology and arrhythmia induction. This highlights the utility of histoanatomically detailed models for investigations of cardiac function, in particular for future patient-specific modeling.

Casero R, Burton RAB, Quinn T, Bollensdorff C, Hales P, Schneider JE, Kohl P, Grau V. 2010. Cardiac valve annulus manual segmentation using computer assisted visual feedback in three-dimensional image data. Conference proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference, pp. 738-741. | Show Abstract

Annulus manual segmentation is an important tool for the study of valve anatomy and physiology, for the four main valves of the heart (mitral, tricuspid, aortic and pulmonary). In this paper we review two traditional manual segmentation approaches: slice-by-slice and interpolating a sparse set of landmarks with a spline curve. We propose a new Spline Tool for the open source software platform Seg3D, that is fast and improves spatial coherence by providing visual feedback of the segmentation in real time. The Spline Tool was tested successfully on 14 rat hearts, on all four valves.

Bordas R, Grau V, Burton RB, Hales P, Schneider JE, Gavaghan D, Kohl P, Rodriguez B. 2010. Integrated approach for the study of anatomical variability in the cardiac Purkinje system: from high resolution MRI to electrophysiology simulation. Conference proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference, pp. 6793-6796. | Show Abstract

The ordered electrical stimulation of the ventricles is achieved by a specialized network of fibres known as the Purkinje system. The gross anatomy and basic functional role of the Purkinje system is well understood. However, very little is known about the detailed anatomy of the Purkinje system, its inter-individual variability and the implications of the variability in ventricular function, in part due to limitations in experimental techniques. In this study, we aim to provide new insight into the inter-individual variability of the free running Purkinje system anatomy and its impact on ventricular electrophysiological function. As a first step towards achieving this aim, high resolution magnetic resonance imaging (MRI) datasets of rat and the rabbit ventricles are obtained and analysed using a novel semi-automatic image processing algorithm for segmentation of the free-running Purkinje system. Segmented geometry from the MRI datasets is used to construct a computational model of the Purkinje system, which is incorporated in to an anatomically-based ventricular geometry to simulate ventricular electrophysiological activity.

Akhtar AM, Schneider JE, Chapman SJ, Jefferson A, Digby JE, Mankia K, Chen Y, McAteer MA, Wood KJ, Choudhury RP. 2010. In vivo quantification of VCAM-1 expression in renal ischemia reperfusion injury using non-invasive magnetic resonance molecular imaging. PLoS One, 5 (9), pp. e12800. | Show Abstract | Read more

RATIONALE AND OBJECTIVE: Vascular cell adhesion molecule-1 (VCAM-1) is upregulated in ischemia reperfusion injury (IRI), persisting after restoration of blood flow. We hypothesized that microparticles of iron oxide targeting VCAM-1 (VCAM-MPIO) would depict "ischemic memory" and enable in vivo assessment of VCAM-1 expression. METHODOLOGY AND FINDINGS: Mice subject to unilateral, transient (30 minutes) renal ischemia and subsequent reperfusion received intravenous VCAM-MPIO (4.5 mg iron/kg body weight). Contrast agent bound rapidly (<30 minutes) in IRI-kidneys and appeared as intensely low signal areas by MRI in vivo. Automated segmentation and quantification yielded MPIO contrast volumes of 5991±354×10(6) µm(3) in IRI vs. 87±7×10(6) µm(3) in kidneys with no surgical intervention (P<0.001); 90±8×10(6) µm(3) in IRI kidneys exposed to control (IgG-MPIO) and 625±80×10(6) µm(3), in IRI kidneys pre-treated with a blocking dose of VCAM-1 antibody (P<0.001). In keeping with quantitative MRI data, VCAM-1 mRNA expression in IRI was 65-fold higher than in kidneys without surgical intervention (3.06±0.63 vs. 0.05±0.02, P<0.001). Indeed VCAM-1 mRNA expression and VCAM-MPIO contrast volume were highly correlated (R(2)=0.901, P<0.01), indicating that quantification of contrast volume reflected renal VCAM-1 transcription. Serial imaging showed VCAM-MPIO accumulation at target within 30 minutes, persisting for ≥90 minutes, while unbound VCAM-MPIO was cleared rapidly from blood, with sequestration by mac-3 positive Kupffer cells in the liver and monocyte/macrophages in the spleen. CONCLUSIONS: (1) VCAM-MPIO detected VCAM-1 expression and defined its 3-dimensional distribution, revealing "ischemic memory" in renal IRI; (2) automated volumetric quantification of VCAM-MPIO accurately reflected tissue levels of VCAM-1 mRNA; and (3) VCAM-MPIO bound rapidly to target with active sequestration of unbound MPIO in the liver and spleen.

Walter T, Shattuck DW, Baldock R, Bastin ME, Carpenter AE, Duce S, Ellenberg J, Fraser A, Hamilton N, Pieper S et al. 2010. Visualization of image data from cells to organisms. Nat Methods, 7 (3 Suppl), pp. S26-S41. | Show Abstract | Read more

Advances in imaging techniques and high-throughput technologies are providing scientists with unprecedented possibilities to visualize internal structures of cells, organs and organisms and to collect systematic image data characterizing genes and proteins on a large scale. To make the best use of these increasingly complex and large image data resources, the scientific community must be provided with methods to query, analyze and crosslink these resources to give an intuitive visual representation of the data. This review gives an overview of existing methods and tools for this purpose and highlights some of their limitations and challenges.

Casero R, Burton RAB, Quinn TA, Bolensdorff C, Hales P, Schneider JE, Kohl P, Grau V. 2010. Towards High-Resolution Cardiac Atlases: Ventricular Anatomy Descriptors for a Standardized Reference Frame STATISTICAL ATLASES AND COMPUTATIONAL MODELS OF THE HEART, 6364 pp. 75-84. | Show Abstract | Read more

Increased resolution in cardiac Magnetic Resonance Imaging (MRI) and growing interest in the effect of small structures in electrophysiology of the heart pose new challenges for cardiac atlases. In this paper we discuss the limitations of current atlas-building models when trying to incorporate cardiac small structure and argue for the need of developing a standard coordinate system for the heart that separates this from the macro-structure common to all individual hearts, in a way analogous to the stereotactic coordinate system from brain atlases. With this goal, we propose a set of methods to obtain two descriptors of the ventricular macro-structure that can be used to build a standardized reference frame: the central curve on the Left Ventricle cavity and the smoothed internal envelope of the Right Ventricle crest (i.e. the curve in the endocardial surface marking the junction between the right ventricular free wall and the septum). © 2010 Springer-Verlag Berlin Heidelberg.

Bohl S, Medway DJ, Schulz-Menger J, Schneider JE, Neubauer S, Lygate CA. 2009. Refined approach for quantification of in vivo ischemia-reperfusion injury in the mouse heart. Am J Physiol Heart Circ Physiol, 297 (6), pp. H2054-H2058. | Show Abstract | Read more

Cardiac ischemia-reperfusion experiments in the mouse are important in vivo models of human disease. Infarct size is a particularly important scientific readout as virtually all cardiocirculatory pathways are affected by it. Therefore, such measurements must be exact and valid. The histological analysis, however, remains technically challenging, and the resulting quality is often unsatisfactory. For this report we have scrutinized each step involved in standard double-staining histology. We have tested published approaches and challenged their practicality. As a result, we propose an improved and streamlined protocol, which consistently yields high-quality histology, thereby minimizing experimental noise and group sizes.

Cleary JO, Price AN, Thomas DL, Scambler PJ, Kyriakopoulou V, McCue K, Schneider JE, Ordidge RJ, Lythgoe MF. 2009. Cardiac phenotyping in ex vivo murine embryos using microMRI. NMR Biomed, 22 (8), pp. 857-866. | Show Abstract | Read more

Microscopic MRI (microMRI) is an emerging technique for high-throughput phenotyping of transgenic mouse embryos, and is capable of visualising abnormalities in cardiac development. To identify cardiac defects in embryos, we have optimised embryo preparation and MR acquisition parameters to maximise image quality and assess the phenotypic changes in chromodomain helicase DNA-binding protein 7 (Chd7) transgenic mice. microMRI methods rely on tissue penetration with a gadolinium chelate contrast agent to reduce tissue T(1), thus improving signal-to-noise ratio (SNR) in rapid gradient echo sequences. We investigated 15.5 days post coitum (dpc) wild-type CD-1 embryos fixed in gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA) solutions for either 3 days (2 and 4 mM) or 2 weeks (2, 4, 8 and 16 mM). To assess penetration of the contrast agent into heart tissue and enable image contrast simulations, T(1) and T(*) (2) were measured in heart and background agarose. Compared to 3-day, 2-week fixation showed reduced mean T(1) in the heart at both 2 and 4 mM concentrations (p < 0.0001), resulting in calculated signal gains of 23% (2 mM) and 29% (4 mM). Using T(1) and T(*) (2) values from 2-week concentrations, computer simulation of heart and background signal, and ex vivo 3D gradient echo imaging, we demonstrated that 2-week fixed embryos in 8 mM Gd-DTPA in combination with optimised parameters (TE/TR/alpha/number of averages: 9 ms/20 ms/60 degrees /7) produced the largest SNR in the heart (23.2 +/- 1.0) and heart chamber contrast-to-noise ratio (CNR) (27.1 +/- 1.6). These optimised parameters were then applied to an MRI screen of embryos heterozygous for the gene Chd7, implicated in coloboma of the eye, heart defects, atresia of the choanae, retardation of growth, genital/urinary abnormalities, ear abnormalities and deafness (CHARGE) syndrome (a condition partly characterised by cardiovascular birth defects in humans). A ventricular septal defect was readily identified in the screen, consistent with the human phenotype.

Young AA, Barnes H, Davison D, Neubauer S, Schneider JE. 2009. Fast left ventricular mass and volume assessment in mice with three-dimensional guide-point modeling. J Magn Reson Imaging, 30 (3), pp. 514-520. | Show Abstract | Read more

PURPOSE: To investigate the accuracy (vs. standard manual analysis) and precision (scan-rescan reproducibility) of three-dimensional guide-point modeling (GPM) for the assessment of left ventricular (LV) function in mice. METHODS: Six male wildtype C57/Bl6 mice (weight 26.2 +/- 1.1 g) were scanned twice, 3 days apart. Each scan was performed twice, at 0.2 mm/pixel with one average and at 0.1 mm/pixel with two averages. The 24 studies were anonymized and analyzed in blinded fashion using GPM and standard manual slice summation. RESULTS: The average error between GPM and standard analysis was 2.3 +/- 5.8 mg in mass, 1.7 +/- 3.2 microL in end-diastolic volume, 2.3 +/- 3.1 microL in end-systolic volume, -2.7 +/- 4.3% in ejection fraction, -0.6 +/- 3.3 microL in stroke volume, and -0.31 +/- 1.56 ml . min(-1) in cardiac output (mean difference +/- SD of differences, n = 24). The average time taken was 8.0 +/- 2.5 minutes for 3D GPM and 48.5 +/- 8.9 minutes for standard analysis (n = 24). Scan-rescan reproducibility results were similar to the standard analysis. No significant differences were found using linear mixed effects modeling in either accuracy or precision between scan resolutions or analysis method. CONCLUSION: 3D GPM enables fast analysis of mouse LV function, with similar accuracy and reproducibility to standard analysis. An image resolution of 0.2 mm/pixel with one average is adequate for LV function studies.

Plank G, Burton RA, Hales P, Bishop M, Mansoori T, Bernabeu MO, Garny A, Prassl AJ, Bollensdorff C, Mason F et al. 2009. Generation of histo-anatomically representative models of the individual heart: tools and application. Philos Trans A Math Phys Eng Sci, 367 (1896), pp. 2257-2292. | Show Abstract | Read more

This paper presents methods to build histo-anatomically detailed individualized cardiac models. The models are based on high-resolution three-dimensional anatomical and/or diffusion tensor magnetic resonance images, combined with serial histological sectioning data, and are used to investigate individualized cardiac function. The current state of the art is reviewed, and its limitations are discussed. We assess the challenges associated with the generation of histo-anatomically representative individualized in silico models of the heart. The entire processing pipeline including image acquisition, image processing, mesh generation, model set-up and execution of computer simulations, and the underlying methods are described. The multifaceted challenges associated with these goals are highlighted, suitable solutions are proposed, and an important application of developed high-resolution structure-function models in elucidating the effect of individual structural heterogeneity upon wavefront dynamics is demonstrated.

Prassl AJ, Kickinger F, Ahammer H, Grau V, Schneider JE, Hofer E, Vigmond EJ, Trayanova NA, Plank G. 2009. Automatically generated, anatomically accurate meshes for cardiac electrophysiology problems. IEEE Trans Biomed Eng, 56 (5), pp. 1318-1330. | Show Abstract | Read more

Significant advancements in imaging technology and the dramatic increase in computer power over the last few years broke the ground for the construction of anatomically realistic models of the heart at an unprecedented level of detail. To effectively make use of high-resolution imaging datasets for modeling purposes, the imaged objects have to be discretized. This procedure is trivial for structured grids. However, to develop generally applicable heart models, unstructured grids are much preferable. In this study, a novel image-based unstructured mesh generation technique is proposed. It uses the dual mesh of an octree applied directly to segmented 3-D image stacks. The method produces conformal, boundary-fitted, and hexahedra-dominant meshes. The algorithm operates fully automatically with no requirements for interactivity and generates accurate volume-preserving representations of arbitrarily complex geometries with smooth surfaces. The method is very well suited for cardiac electrophysiological simulations. In the myocardium, the algorithm minimizes variations in element size, whereas in the surrounding medium, the element size is grown larger with the distance to the myocardial surfaces to reduce the computational burden. The numerical feasibility of the approach is demonstrated by discretizing and solving the monodomain and bidomain equations on the generated grids for two preparations of high experimental relevance, a left ventricular wedge preparation, and a papillary muscle.

Bohl S, Lygate CA, Barnes H, Medway D, Stork LA, Schulz-Menger J, Neubauer S, Schneider JE. 2009. Advanced methods for quantification of infarct size in mice using three-dimensional high-field late gadolinium enhancement MRI. Am J Physiol Heart Circ Physiol, 296 (4), pp. H1200-H1208. | Show Abstract | Read more

Conventional methods to quantify infarct size after myocardial infarction in mice are not ideal, requiring either tissue destruction for histology or relying on nondirect measurements such as wall motion. We therefore implemented a fast, high-resolution method to directly measure infarct size in vivo using three-dimensional (3D) late gadolinium enhancement MRI (3D-LGE). Myocardial T1 relaxation was quantified at 9.4 Tesla in five mice, and reproducibility was tested by repeat imaging after 5 days. In a separate set of healthy and infarcted mice (n = 8 of each), continuous T1 measurements were made following intravenous or intraperitoneal injection of a contrast agent (0.5 micromol/g gadolinium-diethylenetriamine pentaacetic acid). The time course of T1 contrast development between viable and nonviable myocardium was thereby determined, with optimal postinjection imaging windows and inversion times identified. Infarct sizes were quantified using 3D-LGE and compared with triphenyltetrazolium chloride histology on day 1 after infarction (n = 8). Baseline myocardial T1 was highly reproducible: the mean value was 952 +/- 41 ms. T1 contrast peaked earlier after intravenous injection than with intraperitoneal injection; however, contrast between viable and nonviable myocardium was comparable for both routes (P = 0.31), with adequate contrast remaining for at least 60 min postinjection. Excellent correlation was obtained between infarct sizes derived from 3D-LGE and histology (r = 0.91, P = 0.002), and Bland-Altman analysis indicated good agreement free from systematic bias. We have validated an improved 3D MRI method to noninvasively quantify infarct size in mice with unsurpassed spatial resolution and tissue contrast. This method is particularly suited to studies requiring early quantification of initial infarct size, for example, to measure damage before intervention with stem cells.

von zur Muhlen C, Peter K, Ali ZA, Schneider JE, McAteer MA, Neubauer S, Channon KM, Bode C, Choudhury RP. 2009. Visualization of activated platelets by targeted magnetic resonance imaging utilizing conformation-specific antibodies against glycoprotein IIb/IIIa. J Vasc Res, 46 (1), pp. 6-14. | Show Abstract | Read more

Ruptured atherosclerotic plaques, lined with activated platelets, constitute an attractive target for magnetic resonance imaging (MRI). This study evaluated whether microparticles of iron oxide (MPIO) targeting ligand-induced binding sites (LIBS) on the activated conformation of glycoprotein IIb/IIIa could be used to image platelets. MPIO (size: 1 microm) were conjugated to anti-LIBS or control single-chain antibody. Following guidewire injury to mouse femoral artery, platelet adhesion was present after 24 h. Mice were perfused with anti-LIBS-MPIO (or control MPIO) via the left ventricle and 11.7-tesla MRI was performed on femoral arteries ex vivo. A 3D gradient echo sequence attained an isotropic resolution of 25 microm. MPIO binding, quantified by MRI, was 4-fold higher with anti-LIBS-MPIO in comparison to control MPIO (p < 0.01). In histological sections, low signal zones on MRI and MPIO correlated strongly (R(2) = 0.72; p < 0.001), indicating accurate MR quantification. In conclusion, anti-LIBS-MPIO bind to activated platelets in mouse arteries, providing a basis for the use of function-specific single-chain antibody-MPIO conjugates for molecular MRI, and represent the first molecular imaging of a conformational change in a surface receptor. This presents an opportunity to specifically image activated platelets involved in acute atherothrombosis with MRI.

Zhang W, ten Hove M, Schneider JE, Stuckey DJ, Sebag-Montefiore L, Bia BL, Radda GK, Davies KE, Neubauer S, Clarke K. 2008. Abnormal cardiac morphology, function and energy metabolism in the dystrophic mdx mouse: an MRI and MRS study. J Mol Cell Cardiol, 45 (6), pp. 754-760. | Show Abstract | Read more

Patients with muscular dystrophy have abnormal cardiac function and decreased high-energy phosphate metabolism. Here, we have determined whether the 8 month old mdx mouse, an animal model of muscular dystrophy, also has abnormal cardiac function and energetics. In vivo cardiac MRI revealed 33% and 104% larger right ventricular end-diastolic and end-systolic volumes, respectively, and 17% lower right ventricular ejection fractions in mdx mice compared with controls. Evidence of left ventricular diastolic dysfunction included 18% lower peak filling rates in mdx mouse hearts. Abnormal cardiac function was accompanied by necrosis and lower citrate synthase activity in the mdx mouse heart, suggesting decreased mitochondrial content. Decreased mitochondrial numbers were associated with 38% lower phosphocreatine concentration, 22% lower total creatine, 36% higher cytosolic free ADP concentration and 1.3 kJ/mol lower free-energy available from ATP hydrolysis in whole isolated, perfused mdx mouse hearts than in controls. Transsarcolemmal creatine uptake was 12% lower in mdx mouse hearts. We conclude that the absence of dystrophin in adult mdx mouse heart, as in the heart of human patient, is associated with right ventricular dilatation, left ventricular diastolic dysfunction and abnormal energy metabolism.

Carr CA, Stuckey DJ, Tatton L, Tyler DJ, Hale SJ, Sweeney D, Schneider JE, Martin-Rendon E, Radda GK, Harding SE et al. 2008. Bone marrow-derived stromal cells home to and remain in the infarcted rat heart but fail to improve function: an in vivo cine-MRI study. Am J Physiol Heart Circ Physiol, 295 (2), pp. H533-H542. | Show Abstract | Read more

Basic and clinical studies have shown that bone marrow cell therapy can improve cardiac function following infarction. In experimental animals, reported stem cell-mediated changes range from no measurable improvement to the complete restoration of function. In the clinic, however, the average improvement in left ventricular ejection fraction is around 2% to 3%. A possible explanation for the discrepancy between basic and clinical results is that few basic studies have used the magnetic resonance (MR) imaging (MRI) methods that were used in clinical trials for measuring cardiac function. Consequently, we employed cine-MR to determine the effect of bone marrow stromal cells (BMSCs) on cardiac function in rats. Cultured rat BMSCs were characterized using flow cytometry and labeled with iron oxide particles and a fluorescent marker to allow in vivo cell tracking and ex vivo cell identification, respectively. Neither label affected in vitro cell proliferation or differentiation. Rat hearts were infarcted, and BMSCs or control media were injected into the infarct periphery (n = 34) or infused systemically (n = 30). MRI was used to measure cardiac morphology and function and to determine cell distribution for 10 wk after infarction and cell therapy. In vivo MRI, histology, and cell reisolation confirmed successful BMSC delivery and retention within the myocardium throughout the experiment. However, no significant improvement in any measure of cardiac function was observed at any time. We conclude that cultured BMSCs are not the optimal cell population to treat the infarcted heart.

Szumska D, Pieles G, Essalmani R, Bilski M, Mesnard D, Kaur K, Franklyn A, El Omari K, Jefferis J, Bentham J et al. 2008. VACTERL/caudal regression/Currarino syndrome-like malformations in mice with mutation in the proprotein convertase Pcsk5. Genes Dev, 22 (11), pp. 1465-1477. | Show Abstract | Read more

We have identified an ethylnitrosourea (ENU)-induced recessive mouse mutation (Vcc) with a pleiotropic phenotype that includes cardiac, tracheoesophageal, anorectal, anteroposterior patterning defects, exomphalos, hindlimb hypoplasia, a presacral mass, renal and palatal agenesis, and pulmonary hypoplasia. It results from a C470R mutation in the proprotein convertase PCSK5 (PC5/6). Compound mutants (Pcsk5(Vcc/null)) completely recapitulate the Pcsk5(Vcc/Vcc) phenotype, as does an epiblast-specific conditional deletion of Pcsk5. The C470R mutation ablates a disulfide bond in the P domain, and blocks export from the endoplasmic reticulum and proprotein convertase activity. We show that GDF11 is cleaved and activated by PCSK5A, but not by PCSK5A-C470R, and that Gdf11-deficient embryos, in addition to having anteroposterior patterning defects and renal and palatal agenesis, also have a presacral mass, anorectal malformation, and exomphalos. Pcsk5 mutation results in abnormal expression of several paralogous Hox genes (Hoxa, Hoxc, and Hoxd), and of Mnx1 (Hlxb9). These include known Gdf11 targets, and are necessary for caudal embryo development. We identified nonsynonymous mutations in PCSK5 in patients with VACTERL (vertebral, anorectal, cardiac, tracheoesophageal, renal, limb malformation OMIM 192350) and caudal regression syndrome, the phenotypic features of which resemble the mouse mutation. We propose that Pcsk5, at least in part via GDF11, coordinately regulates caudal Hox paralogs, to control anteroposterior patterning, nephrogenesis, skeletal, and anorectal development.

Schneider JE, Lanz T, Barnes H, Medway D, Stork LA, Lygate CA, Smart S, Griswold MA, Neubauer S. 2008. Ultra-fast and accurate assessment of cardiac function in rats using accelerated MRI at 9.4 Tesla. Magn Reson Med, 59 (3), pp. 636-641. | Show Abstract | Read more

MRI can accurately and reproducibly assess cardiac function in rodents but requires relatively long imaging times. Therefore, parallel imaging techniques using a 4-element RF-coil array and MR sequences for cardiac MRI in rats were implemented at ultra-high magnetic fields (9.4 Tesla [T]). The hypothesis that these developments would result in a major reduction in imaging time without loss of accuracy was tested on female Wistar rats under isoflurane anesthesia. High-resolution, contiguous short-axis slices (thickness 1.5 mm) were acquired covering the entire heart. Two interleaved data sets (i) with the volume coil (eight averages) and (ii) with the four-element coil array (one average) were obtained. In addition, two-, three-, and fourfold accelerated data sets were generated through postprocessing of the coil array data, followed by a TGRAPPA reconstruction, resulting in five data sets per rat (in-plane voxel size 100 x 100 microm). Using a single blinded operator, excellent agreement was obtained between volume coil (acquisition time: 88 min) and the fourfold accelerated (<3 min) data sets (e.g., LV mass 436 +/- 21 mg vs 433 +/- 19 mg; ejection fraction 74 +/- 5% vs 75 +/- 4%). This finding demonstrates that it is possible to complete a rat cine-MRI study under 3 min with low variability and without losing temporal or spatial resolution, making high throughput screening programs feasible.

McAteer MA, Schneider JE, Ali ZA, Warrick N, Bursill CA, von zur Muhlen C, Greaves DR, Neubauer S, Channon KM, Choudhury RP. 2008. Magnetic resonance imaging of endothelial adhesion molecules in mouse atherosclerosis using dual-targeted microparticles of iron oxide. Arterioscler Thromb Vasc Biol, 28 (1), pp. 77-83. | Show Abstract | Read more

OBJECTIVE: Microparticles of iron oxide (MPIO) distort magnetic field creating marked contrast effects far exceeding their physical size. We hypothesized that antibody-conjugated MPIO would enable magnetic resonance imaging (MRI) of endothelial cell adhesion molecules in mouse atherosclerosis. METHODS AND RESULTS: MPIO (4.5 microm) were conjugated to monoclonal antibodies against vascular cell adhesion molecule-1 (VCAM-MPIO) or P-selectin (P-selectin-MPIO). In vitro, VCAM-MPIO bound, in dose-dependent manner, to tumor necrosis factor (TNF)-alpha stimulated sEND-1 endothelial cells, as quantified by light microscopy (R2=0.94, P=0.03) and by MRI (R2=0.98, P=0.01). VCAM-MPIO binding was blocked by preincubation with soluble VCAM-1. To mimic leukocyte binding, MPIO targeting both VCAM-1 and P-selectin were administered in apolipoprotein E-/- mice. By light microscopy, dual-targeted MPIO binding to endothelium overlying aortic root atherosclerosis was 5- to 7-fold more than P-selectin-MPIO (P<0.05) or VCAM-MPIO (P<0.01) alone. Dual-targeted MPIO, injected intravenously in vivo bound aortic root endothelium and were quantifiable by MRI ex vivo (3.5-fold increase versus control; P<0.01). MPIO were well-tolerated in vivo, with sequestration in the spleen after 24 hours. CONCLUSIONS: Dual-ligand MPIO bound to endothelium over atherosclerosis in vivo, under flow conditions. MPIO may provide a functional MRI probe for detecting endothelial-specific markers in a range of vascular pathologies.

Schneider JE, Stork LA, Bell JT, ten Hove M, Isbrandt D, Clarke K, Watkins H, Lygate CA, Neubauer S. 2008. Cardiac structure and function during ageing in energetically compromised Guanidinoacetate N-methyltransferase (GAMT)-knockout mice - a one year longitudinal MRI study. J Cardiovasc Magn Reson, 10 (1), pp. 9. | Show Abstract | Read more

BACKGROUND: High-resolution magnetic resonance imaging (cine-MRI) is well suited for determining global cardiac function longitudinally in genetically or surgically manipulated mice, but in practice it is seldom used to its full potential. In this study, male and female guanidinoacetate N-methyltransferase (GAMT) knockout, and wild type littermate mice were subjected to a longitudinal cine-MRI study at four time points over the course of one year. GAMT is an essential enzyme in creatine biosynthesis, such that GAMT deficient mice are entirely creatine-free. Since creatine plays an important role in the buffering and transfer of high-energy phosphate bonds in the heart, it was hypothesized that lack of creatine would be detrimental for resting cardiac performance during ageing. METHODS: Measurements of cardiac structure (left ventricular mass and volumes) and function (ejection fraction, stroke volume, cardiac output) were obtained using high-resolution cine-MRI at 9.4 T under isoflurane anaesthesia. RESULTS: There were no physiologically significant differences in cardiac function between wild type and GAMT knockout mice at any time point for male or female groups, or for both combined (for example ejection fraction: 6 weeks (KO vs. WT): 70 +/- 6% vs. 65 +/- 7%; 4 months: 70 +/- 6% vs. 62 +/- 8%; 8 months: 62 +/- 11% vs. 62 +/- 6%; 12 months: 61 +/- 7% vs. 59 +/- 11%, respectively). CONCLUSION: These findings suggest the presence of comprehensive adaptations in the knockout mice that can compensate for a lack of creatine. Furthermore, this study clearly demonstrates the power of cine-MRI for accurate non-invasive, serial cardiac measurements. Cardiac growth curves could easily be defined for each group, in the same set of animals for all time points, providing improved statistical power, and substantially reducing the number of mice required to conduct such a study. This technique should be eminently useful for following changes of cardiac structure and function during ageing.

McAteer MA, Sibson NR, von Zur Muhlen C, Schneider JE, Lowe AS, Warrick N, Channon KM, Anthony DC, Choudhury RP. 2007. In vivo magnetic resonance imaging of acute brain inflammation using microparticles of iron oxide. Nat Med, 13 (10), pp. 1253-1258. | Show Abstract | Read more

Multiple sclerosis is a disease of the central nervous system that is associated with leukocyte recruitment and subsequent inflammation, demyelination and axonal loss. Endothelial vascular cell adhesion molecule-1 (VCAM-1) and its ligand, alpha4beta1 integrin, are key mediators of leukocyte recruitment, and selective inhibitors that bind to the alpha4 subunit of alpha4beta1 substantially reduce clinical relapse in multiple sclerosis. Urgently needed is a molecular imaging technique to accelerate diagnosis, to quantify disease activity and to guide specific therapy. Here we report in vivo detection of VCAM-1 in acute brain inflammation, by magnetic resonance imaging in a mouse model, at a time when pathology is otherwise undetectable. Antibody-conjugated microparticles carrying a large amount of iron oxide provide potent, quantifiable contrast effects that delineate the architecture of activated cerebral blood vessels. Their rapid clearance from blood results in minimal background contrast. This technology is adaptable to monitor the expression of endovascular molecules in vivo in various pathologies.

Macdonald ST, Bamforth SD, Farthing CR, Franklyn AJ, Broadbent C, Schneider J, Schwartz RJ, Saga Y, Bhattacharya S. 2007. Epiblastic Cited2 function explains pleiotropy and penetrance of cardiac malformation resulting from its deficiency EUROPEAN HEART JOURNAL, 28 pp. 123-123.

Pieles G, Geyer SH, Szumska D, Schneider J, Neubauer S, Clarke K, Dorfmeister K, Franklyn A, Brown SD, Bhattacharya S, Weninger WJ. 2007. microMRI-HREM pipeline for high-throughput, high-resolution phenotyping of murine embryos. J Anat, 211 (1), pp. 132-137. | Show Abstract | Read more

Rapid and precise phenotyping analysis of large numbers of wild-type and mutant mouse embryos is essential for characterizing the genetic and epigenetic factors regulating embryogenesis. We present a novel methodology that permits precise high-throughput screening of the phenotype of embryos with both targeted and randomly generated mutations. To demonstrate the potential of this methodology we show embryo phenotyping results produced in a large-scale ENU-mutagenesis study. In essence this represents an analysis pipeline, which starts with simultaneous micro-magentic resonance imaging (microMRI) screening (voxel size: 25.4 x 25.4 x 24.4 microm) of 32 embryos in one run. Embryos with an indistinct phenotype are then cut into parts and suspect organs and structures are analysed with HREM (high-resolution episcopic microscopy). HREM is an imaging technique that employs 'positive' eosin staining and episcopic imaging for generating three-dimensional (3D) high-resolution (voxel size: 1.07 x 1.07 x 2 microm) digital data of near histological contrast and quality. The results show that our method guarantees the rapid availability of comprehensive phenotype information for high numbers of embryos in, if necessary, histological quality and detail. The combination of high-throughput microMRI with HREM provides an alternative screening pipeline with advantages over existing 3D phenotype screening methods as well as traditional histology. Thus, the microMRI-HREM phenotype analysis pipeline recommends itself as a routine tool for analysing the phenotype of transgenic and mutant embryos.

Liang X, Sun Y, Schneider J, Ding JH, Cheng H, Ye M, Bhattacharya S, Rearden A, Evans S, Chen J. 2007. Pinch1 is required for normal development of cranial and cardiac neural crest-derived structures. Circ Res, 100 (4), pp. 527-535. | Show Abstract | Read more

Pinch1, an adaptor protein composed of 5 LIM domains, has been suggested to play an important role in multiple cellular processes. We found that Pinch1 is highly expressed in neural crest cells and their derivatives. To examine the requirement for Pinch1 in neural crest development, we generated neural crest conditional Pinch1 knockout mice using the Wnt1-Cre/loxP system. Neural crest conditional Pinch1 mutant embryos die perinatally from severe cardiovascular defects with an unusual aneurysmal common arterial trunk. Pinch1 mutants also exhibit multiple deficiencies in cranial neural crest-derived structures. Fate mapping demonstrated that initial migration of neural crest cells to the pharyngeal arch region occurs normally in the mutant embryos. However, in the cardiac outflow tract of mutants, neural crest cells exhibited hyperplasia and failed to differentiate into smooth muscle. Markedly increased apoptosis is observed in outflow tract cushions of mutants between embryonic days 11.5 and 13.5, likely contributing to the observed defects in cushion/valve remodeling and ventricular septation. Expression of transforming growth factor-beta(2), which plays a crucial role in outflow tract development, was decreased or absent in the outflow tract of the mutants. The decrease in transforming growth factor-beta(2) expression preceded neural crest cell death. Together, our results demonstrate that Pinch1 plays an essential role in neural crest development, perhaps in part through transforming growth factor-beta signaling.

Panagia M, Schneider JE, Brown B, Cole MA, Clarke K. 2007. Abnormal function and glucose metabolism in the type-2 diabetic db/db mouse heart. Can J Physiol Pharmacol, 85 (3-4), pp. 289-294. | Show Abstract | Read more

This study examined cardiac function and glucose metabolism in the 6-month-old db/db mouse, a model of type-2 diabetes. Cine magnetic resonance spectroscopy (MRI) was used to measure cardiac function in vivo. The db/db mice had decreased heart rates (17%, p<0.01) and stroke volumes (21%, p<0.05) that resulted in lower cardiac output (35%, p<0.01) than controls. Although there was no difference in ejection fraction between the 2 groups, db/db mouse hearts had a 35% lower maximum rate of ejection (p<0.01) than controls. In a protocol designed to assess maximal insulin-independent glucose uptake, hearts were isolated and perfused in Langendorff mode and subjected to 0.75 mL.min(-1).(g wet mass)(-1) low flow ischemia for 32 min. Glucose uptake during ischemia was 21% lower than in controls, and post-ischemic recovery of cardiac function was decreased by 30% in db/db mouse hearts (p<0.05). Total cardiac GLUT 4 protein was 56% lower (p<0.01) in db/db mice than in controls. In summary, the db/db mouse has abnormal left ventricular function in vivo, with impaired glucose uptake during ischemia, leading to increased myocardial damage.

Goodyer CE, Grau V, Mansoori T, Schneider JE, Brodlie KW, Kohl P, IEEE. 2007. 3D visualization of cardiac anatomical MRI data with para-cellular resolution 2007 ANNUAL INTERNATIONAL CONFERENCE OF THE IEEE ENGINEERING IN MEDICINE AND BIOLOGY SOCIETY, VOLS 1-16, pp. 147-+. | Show Abstract | Read more

Advances in research and clinical techniques are providing increasing quantities of data at improved spatio-temporal resolution. It is therefore imperative to develop matching approaches for efficient analysis and intuitive presentation of this data. Using the example of advanced magnetic resonance imaging, this article will illustrate the challenges involved in computational reconstruction and interactive visualization of the three-dimensional cardiac anatomy, based on magnetic resonance imaging data with para-cellular resolution. © 2007 IEEE.

Catchpole AF, Carr CA, Schneider JE, Tyler DJ, Stuckey DJ, Clarke K. 2006. Inflammation and insulin resistance in the acutely infarcted rat heart CARDIOVASCULAR DRUGS AND THERAPY, 20 (6), pp. 401-402.

Burton RA, Plank G, Schneider JE, Grau V, Ahammer H, Keeling SL, Lee J, Smith NP, Gavaghan D, Trayanova N, Kohl P. 2006. Three-dimensional models of individual cardiac histoanatomy: tools and challenges. Ann N Y Acad Sci, 1080 (1), pp. 301-319. | Show Abstract | Read more

There is a need for, and utility in, the acquisition of data sets of cardiac histoanatomy, with the vision of reconstructing individual hearts on the basis of noninvasive imaging, such as MRI, enriched by reference to detailed atlases of serial histology obtained from representative samples. These data sets would be useful not only as a repository of knowledge regarding the specifics of cardiac histoanatomy, but could form the basis for generation of individualized high-resolution cardiac structure-function models. The current article presents a step in this general direction: it illustrates how whole-heart noninvasive imaging can be combined with whole-heart histology in an approach to achieve automated construction of histoanatomically detailed models of cardiac 3D structure and function at hitherto unprecedented resolution and accuracy (based on 26.4 x 26.4 x 24.4 microm MRI voxel size, and enriched by histological detail). It provides an overview of the tools used in this quest and outlines challenges posed by the approach in the light of applications that may benefit from the availability of such data and tools.

Hart AW, Morgan JE, Schneider J, West K, McKie L, Bhattacharya S, Jackson IJ, Cross SH. 2006. Cardiac malformations and midline skeletal defects in mice lacking filamin A. Hum Mol Genet, 15 (16), pp. 2457-2467. | Show Abstract | Read more

The X-linked gene filamin A (Flna) encodes a widely expressed actin-binding protein that crosslinks actin into orthogonal networks and interacts with a variety of other proteins including membrane proteins, integrins, transmembrane receptor complexes and second messengers, thus forming an important intracellular signalling scaffold. Heterozygous loss of function of human FLNA causes periventricular nodular heterotopia in females and is generally lethal (cause unknown) in hemizygous males. Missense FLNA mutations underlie a spectrum of disorders affecting both sexes that feature skeletal dysplasia accompanied by a variety of other abnormalities. Dilp2 is an X-linked male-lethal mouse mutation that was induced by N-ethyl-N-nitrosourea. We report here that Dilp2 is caused by a T-to-A transversion that converts a tyrosine codon to a stop codon in the Flna gene (Y2388X), leading to absence of the Flna protein and male lethality because of incomplete septation of the outflow tract of the heart, which produces common arterial trunk. A proportion of both male and female mutant mice have other cardiac defects including ventricular septal defect. In addition, mutant males have midline fusion defects manifesting as sternum and palate abnormalities. Carrier females exhibit milder sternum and palate defects and misshapen pupils. These results define crucial roles for Flna in development, demonstrate that X-linked male lethal mutations can be recovered from ENU mutagenesis screens and suggest possible explanations for lethality of human males hemizygous for null alleles of FLNA.

Stuckey DJ, Carr CA, Martin-Rendon E, Tyler DJ, Willmott C, Cassidy PJ, Hale SJ, Schneider JE, Tatton L, Harding SE et al. 2006. Iron particles for noninvasive monitoring of bone marrow stromal cell engraftment into, and isolation of viable engrafted donor cells from, the heart. Stem Cells, 24 (8), pp. 1968-1975. | Show Abstract | Read more

Stem cells offer a promising approach to the treatment of myocardial infarction and prevention of heart failure. We have used iron labeling of bone marrow stromal cells (BMSCs) to noninvasively track cell location in the infarcted rat heart over 16 weeks using cine-magnetic resonance imaging (cine-MRI) and to isolate the BMSCs from the grafted hearts using the magnetic properties of the donor cells. BMSCs were isolated from rat bone marrow, characterized by flow cytometry, transduced with lentiviral vectors expressing green fluorescent protein (GFP), and labeled with iron particles. BMSCs were injected into the infarct periphery immediately following coronary artery ligation, and rat hearts were imaged at 1, 4, 10, and 16 weeks postinfarction. Signal voids caused by the iron particles in the BMSCs were detected in all rats at all time points. In mildly infarcted hearts, the volume of the signal void decreased over the 16 weeks, whereas the signal void volume did not decrease significantly in severely infarcted hearts. High-resolution three-dimensional magnetic resonance (MR) microscopy identified hypointense regions at the same position as in vivo. Donor cells containing iron particles and expressing GFP were identified in MR-targeted heart sections after magnetic cell separation from digested hearts. In conclusion, MRI can be used to track cells labeled with iron particles in damaged tissue for at least 16 weeks after injection and to guide tissue sectioning by accurately identifying regions of cell engraftment. The magnetic properties of the iron-labeled donor cells can be used for their isolation from host tissue to enable further characterization.

Davies JK, Wells DJ, Liu K, Whitrow HR, Daniel TD, Grignani R, Lygate CA, Schneider JE, Noël G, Watkins H, Carling D. 2006. Characterization of the role of gamma2 R531G mutation in AMP-activated protein kinase in cardiac hypertrophy and Wolff-Parkinson-White syndrome. Am J Physiol Heart Circ Physiol, 290 (5), pp. H1942-H1951. | Show Abstract | Read more

AMP-activated protein kinase (AMPK) is the downstream component of a protein kinase cascade that plays a key role in the regulation of energy metabolism. In humans, mutations in the gamma2-subunit of AMPK cause cardiac hypertrophy associated with Wolff-Parkinson-White syndrome, characterized by ventricular preexcitation. The effect of these mutations on AMPK activity and in development of the disease is enigmatic. Here we report that transgenic mice with cardiac-specific expression of gamma2 harboring a mutation of arginine residue 531 to glycine (RG-TG) develop a striking cardiac phenotype by 4 wk of age, including hypertrophy, impaired contractile function, electrical conduction abnormalities, and marked glycogen accumulation. At this stage, AMPK activity isolated from hearts of RG-TG mice was almost completely abolished but could be restored after phosphorylation by an upstream AMPK kinase. At 1 wk of age, there was no detectable evidence of a cardiac phenotype, and AMPK activity in RG-TG hearts was similar to that in nontransgenic, control mice. We propose that mutations in gamma2 lead to suppression of total cardiac AMPK activity secondary to increased glycogen accumulation. The subsequent decrease in AMPK activity provides a mechanism that may explain the development of cardiac hypertrophy in this model.

Schneider JE, Wiesmann F, Lygate CA, Neubauer S. 2006. How to perform an accurate assessment of cardiac function in mice using high-resolution magnetic resonance imaging. J Cardiovasc Magn Reson, 8 (5), pp. 693-701. | Show Abstract | Read more

High-resolution magnetic resonance cine imaging (cine-MRI) is a method that allows for a non-invasive assessment of left ventricular function and mass. To perform this quantitation, hearts are imaged from the base to the apex by a stack of two-dimensional images. Thus, analysis of myocardial mass and function by cine-MRI does not rely on geometric assumptions. Geometric and functional parameters, such as end-diastolic volume (EDV), end-systolic volume (ESV) or ejection fraction (EF), are obtained by subsequent image segmentation of the respective cine frames in each slice. While this technique has been well established in clinical practice, it is now rapidly becoming the reference method in experimental cardiovascular MRI for accurate quantification of cardiac parameters, thereby aiding the phenotyping of the increasing number of transgenic and surgical mouse models. However, accurate measurement of cardiac functional parameters requires the images to be acquired in short-axis orientation of the heart, which can be difficult to define, particularly in animals with diseased hearts. Furthermore, data analysis can be the source of a systematic error, mainly for myocardial mass measurement. Here, we describe a protocol that allows for a quick and reproducible approach of obtaining the relevant cardiac views for cine-MRI, and we explain how an accurate experimental image analysis can be performed.

Tyler DJ, Lygate CA, Schneider JE, Cassidy PJ, Neubauer S, Clarke K. 2006. CINE-MR imaging of the normal and infarcted rat heart using an 11.7 T vertical bore MR system. J Cardiovasc Magn Reson, 8 (2), pp. 327-333. | Show Abstract | Read more

MR imaging is uniquely placed to non-invasively study rodent cardiac structure and function. High-field MR scanners commonly have a vertical bore, and the purpose of this work was to demonstrate CINE-MR imaging in normal and infarcted rat hearts after determining hemodynamic stability when positioned vertically for imaging. Optimisation of imaging parameters was carried out prior to assessment of cardiac function in a group of normal and infarcted rat hearts. Rat hemodynamics were unaltered when vertical for 90 minutes, compared with horizontal measurements and rat cardiac parameters were measured accurately and reproducibly with our optimized CINE-MR protocol. A flip angle of 17.5 degrees was shown to provide optimal contrast for the assessment of structure and function, and, in contrast to our findings in mice, respiratory gating was not found to be essential. Hence, we conclude that vertical bore MR systems can be used to measure in vivo cardiac function in normal and infarcted rat hearts.

Lygate CA, Schneider JE, Hulbert K, ten Hove M, Sebag-Montefiore LM, Cassidy PJ, Clarke K, Neubauer S. 2006. Serial high resolution 3D-MRI after aortic banding in mice: band internalization is a source of variability in the hypertrophic response. Basic Res Cardiol, 101 (1), pp. 8-16. | Show Abstract | Read more

Transverse aortic constriction (TAC) is used as a model of left ventricular hypertrophy and failure; however, there is extensive variability in the hypertrophic response. In 43 mice that underwent TAC with a 7-0 polypropylene suture, 13 were identified by echocardiography with initial LV hypertrophy that halted or regressed over time. Post-mortem examination on 7 of these mice found the constricting band to be intact, but partially internalized into the aortic lumen, allowing blood flow around the stenosis. To confirm this prospectively in vivo we then followed 12 mice after TAC for 6 weeks, using a new high resolution 3D-MRI method to measure minimal aortic arch cross-sectional area (CSA). Three of the 12 mice developed a significantly increased aortic CSA (0.31 +/- 0.15 on day 2 vs. 1.11 +/- 0.29 mm2 on day 42; P < 0.05), which was independently confirmed by dissection. These mice had internalized part of the band within the aortic lumen and, by week 6, showed significantly less LV hypertrophy and better systolic function. Nine of the 12 mice showed no change in aortic CSA. Band internalization could be prevented when two banding sutures were placed side-by-side (n = 10). This is the first observation that a significant subset of animals following TAC bypass the stenosis resulting in partial regression of hypertrophy.

Wallis J, Lygate CA, Fischer A, ten Hove M, Schneider JE, Sebag-Montefiore L, Dawson D, Hulbert K, Zhang W, Zhang MH et al. 2005. Supranormal myocardial creatine and phosphocreatine concentrations lead to cardiac hypertrophy and heart failure: insights from creatine transporter-overexpressing transgenic mice. Circulation, 112 (20), pp. 3131-3139. | Show Abstract | Read more

BACKGROUND: Heart failure is associated with deranged cardiac energy metabolism, including reductions of creatine and phosphocreatine. Interventions that increase myocardial high-energy phosphate stores have been proposed as a strategy for treatment of heart failure. Previously, it has not been possible to increase myocardial creatine and phosphocreatine concentrations to supranormal levels because they are subject to tight regulation by the sarcolemmal creatine transporter (CrT). METHODS AND RESULTS: We therefore created 2 transgenic mouse lines overexpressing the myocardial creatine transporter (CrT-OE). Compared with wild-type (WT) littermate controls, total creatine (by high-performance liquid chromatography) was increased in CrT-OE hearts (66+/-6 nmol/mg protein in WT versus 133+/-52 nmol/mg protein in CrT-OE). Phosphocreatine levels (by 31P magnetic resonance spectroscopy) were also increased but to a lesser extent. Surprisingly, CrT-OE mice developed left ventricular (LV) dilatation (LV end-diastolic volume: 21.5+/-4.3 microL in WT versus 33.1+/-9.6 microL in CrT-OE; P=0.002), substantial LV dysfunction (ejection fraction: 64+/-9% in WT versus 49+/-13% in CrT-OE; range, 22% to 70%; P=0.003), and LV hypertrophy (by 3-dimensional echocardiography and magnetic resonance imaging). Myocardial creatine content correlated closely with LV end-diastolic volume (r=0.51, P=0.02), ejection fraction (r=-0.74, P=0.0002), LV weight (r=0.59, P=0.006), LV end-diastolic pressure (r=0.52, P=0.02), and dP/dt(max) (r=-0.69, P=0.0008). Despite increased creatine and phosphocreatine levels, CrT-OE hearts showed energetic impairment, with increased free ADP concentrations and reduced free-energy change levels. CONCLUSIONS: Overexpression of the CrT leads to supranormal levels of myocardial creatine and phosphocreatine, but the heart is incapable of keeping the augmented creatine pool adequately phosphorylated, resulting in increased free ADP levels, LV hypertrophy, and dysfunction. Our data demonstrate that a disturbance of the CrT-mediated tight regulation of cardiac energy metabolism has deleterious functional consequences. These findings caution against the uncritical use of creatine as a therapeutic agent in heart disease.

Bogani D, Willoughby C, Davies J, Kaur K, Mirza G, Paudyal A, Haines H, McKeone R, Cadman M, Pieles G et al. 2005. Dissecting the genetic complexity of human 6p deletion syndromes by using a region-specific, phenotype-driven mouse screen. Proc Natl Acad Sci U S A, 102 (35), pp. 12477-12482. | Show Abstract | Read more

Monosomy of the human chromosome 6p terminal region results in a variety of congenital malformations that include brain, craniofacial, and organogenesis abnormalities. To examine the genetic basis of these phenotypes, we have carried out an unbiased functional analysis of the syntenic region of the mouse genome (proximal Mmu13). A genetic screen for recessive mutations in this region recovered thirteen lines with phenotypes relevant to a variety of clinical conditions. These include two loci that cause holoprosencephaly, two that underlie anophthalmia, one of which also contributes to other craniofacial abnormalities such as microcephaly, agnathia, and palatogenesis defects, and one locus responsible for developmental heart and kidney defects. Analysis of heterozygous carriers of these mutations shows that a high proportion of these loci manifest with behavioral activity and sensorimotor deficits in the heterozygous state. This finding argues for the systematic, reciprocal phenotypic assessment of dominant and recessive mouse mutants. In addition to providing a resource of single gene mutants that model 6p-associated disorders, the work reveals unsuspected genetic complexity at this region. In particular, many of the phenotypes associated with 6p deletions can be elicited by mutation in one of a number of genes. This finding implies that phenotypes associated with contiguous gene deletion syndromes can result not only from dosage sensitivity of one gene in the region but also from the combined effect of monosomy for multiple genes that function within the same biological process.

ten Hove M, Lygate CA, Fischer A, Schneider JE, Sang AE, Hulbert K, Sebag-Montefiore L, Watkins H, Clarke K, Isbrandt D et al. 2005. Reduced inotropic reserve and increased susceptibility to cardiac ischemia/reperfusion injury in phosphocreatine-deficient guanidinoacetate-N-methyltransferase-knockout mice. Circulation, 111 (19), pp. 2477-2485. | Show Abstract | Read more

BACKGROUND: The role of the creatine kinase (CK)/phosphocreatine (PCr) energy buffer and transport system in heart remains unclear. Guanidinoacetate-N-methyltransferase-knockout (GAMT-/-) mice represent a new model of profoundly altered cardiac energetics, showing undetectable levels of PCr and creatine and accumulation of the precursor (phospho-)guanidinoacetate (P-GA). To characterize the role of a substantially impaired CK/PCr system in heart, we studied the cardiac phenotype of wild-type (WT) and GAMT-/- mice. METHODS AND RESULTS: GAMT-/- mice did not show cardiac hypertrophy (myocyte cross-sectional areas, hypertrophy markers atrial natriuretic factor and beta-myosin heavy chain). Systolic and diastolic function, measured invasively (left ventricular conductance catheter) and noninvasively (MRI), were similar for WT and GAMT-/- mice. However, during inotropic stimulation with dobutamine, preload-recruitable stroke work failed to reach maximal levels of performance in GAMT-/- hearts (101+/-8 mm Hg in WT versus 59+/-7 mm Hg in GAMT-/-; P<0.05). (31)P-MR spectroscopy experiments showed that during inotropic stimulation, isolated WT hearts utilized PCr, whereas isolated GAMT-/- hearts utilized P-GA. During ischemia/reperfusion, GAMT-/- hearts showed markedly impaired recovery of systolic (24% versus 53% rate pressure product recovery; P<0.05) and diastolic function (eg, left ventricular end-diastolic pressure 23+/-9 in WT and 51+/-5 mm Hg in GAMT-/- during reperfusion; P<0.05) and incomplete resynthesis of P-GA. CONCLUSIONS: GAMT-/- mice do not develop hypertrophy and show normal cardiac function at low workload, suggesting that a fully functional CK/PCr system is not essential under resting conditions. However, when acutely stressed by inotropic stimulation or ischemia/reperfusion, GAMT-/- mice exhibit a markedly abnormal phenotype, demonstrating that an intact, high-capacity CK/PCr system is required for situations of increased cardiac work or acute stress.

Wilding JR, Schneider JE, Sang AE, Davies KE, Neubauer S, Clarke K. 2005. Dystrophin- and MLP-deficient mouse hearts: marked differences in morphology and function, but similar accumulation of cytoskeletal proteins. FASEB J, 19 (1), pp. 79-81. | Show Abstract | Read more

In humans, cytoskeletal dystrophin and muscle LIM protein (MLP) gene mutations can cause dilated cardiomyopathy, yet these mutations may have different effects in mice, owing to increased accumulation of other, compensatory cytoskeletal proteins. Consequently, we characterized left-ventricular (LV) morphology and function in vivo using high-resolution cine-magnetic resonance imaging (MRI) in 2- to 3-month old dystrophin-deficient (mdx) and MLP-null mice, and their respective controls. LV passive stiffness was assessed in isolated, perfused hearts, and cytoskeletal protein levels were determined using Western blot analyses. In mdx mouse hearts, LV-to-body weight ratio, cavity volume, ejection fraction, stroke volume, and cardiac output were normal. However, MLP-null mouse hearts had 1.2-fold higher LV-to-body weight ratios (P<0.01), 1.5-fold higher end-diastolic volumes (P<0.01), and decreased ejection fraction compared with controls (25% vs. 66%, respectively, P<0.01), indicating dilated cardiomyopathy and heart failure. In both models, isolated, perfused heart end-diastolic pressure-volume relationships and passive left-ventricular stiffness were normal. Hearts from both models accumulated desmin and beta-tubulin, mdx mouse hearts accumulated utrophin and MLP, and MLP-null mouse hearts accumulated dystrophin and syncoilin. Although the increase in MLP and utrophin in the mdx mouse heart was able to compensate for the loss of dystrophin, accumulation of desmin, syncoilin and dystrophin were unable to compensate for the loss of MLP, resulting in heart failure.

Schneider JE, Hulbert KJ, Lygate CA, Ten Hove M, Cassidy PJ, Clarke K, Neubauer S. 2004. Long-term stability of cardiac function in normal and chronically failing mouse hearts in a vertical-bore MR system. MAGMA, 17 (3-6), pp. 162-169. | Show Abstract | Read more

We previously demonstrated stability of ventricular volumes and cardiac function in normal and in chronically failing mouse hearts in MR systems with a vertical-bore magnet for up to 1 h. However, in order to exploit the benefits of an increased magnetic field strength of these MR systems in more time-consuming studies required by, for example MR spectroscopy, we investigated whether cardiac function and ventricular volumes of healthy and infarcted mice would be affected in vertical position over a prolonged period. We applied high-resolution MR cine imaging on an 11.7 T vertical MR system to monitor cardiac functional parameters of normal and chronically failing mouse hearts over a period of 3 h in an upright position, with a temporal resolution of < or =15 min. We monitored left-ventricular volumes and cardiac functional parameters in both groups. In normal mice, we detected a decrease of left-ventricular end-systolic volumes by 8 microl and an approximately 23% increase of ejection fraction over time indicating a small but detectable degree of orthostatic dysregulation. Observed changes were more pronounced in mice with heart failure. Despite significant changes in left-ventricular volumes and function, absolute values measured for all functional cardiac parameters are consistent with near-physiological conditions. Thus, mice can be studied in high-field MR systems positioned vertically for 3 h.

Schneider JE, McAteer MA, Tyler DJ, Clarke K, Channon KM, Choudhury RP, Neubauer S. 2004. High-resolution, multicontrast three-dimensional-MRI characterizes atherosclerotic plaque composition in ApoE-/- mice ex vivo. J Magn Reson Imaging, 20 (6), pp. 981-989. | Show Abstract | Read more

PURPOSE: To systematically investigate intrinsic MR contrast mechanisms that would facilitate plaque characterization and quantification in the aortic root and brachiocephalic artery of ApoE-/- mice ex vivo. MATERIALS AND METHODS: To establish unambiguous MR parameters for routinely analyzing atherosclerotic plaque ex vivo at 11.7 T, relaxation times of plaque components were quantitatively assessed. Magnetization transfer and lipid-proton three-dimensional MR imaging was investigated for visualization of collagen- and lipid-rich plaque regions, respectively. A three-dimensional multiecho sequence with a spatial resolution of 47 x 47 x 63 microm was implemented providing a variable degree of T2-weighting. RESULTS: Relaxation time measurements showed clear tissue heterogeneity between atherosclerotic plaque components in the T2-values, but similar T1-values at 11.7 T (T1/T2 mean +/- SD; cellular plaque component: 1.2 +/- 0.3 seconds/26.3 +/- 0.4 msec; fibrofatty plaque component: 1.1 +/- 0.2 seconds/13.7 +/- 2.0 msec). The three-dimensional multiecho sequence allowed the calculation of the intrinsic proton density and T2-maps. The sum of the multiecho data provided strong T2-weighting that facilitated quantification of various components of atherosclerotic plaque in the mouse aortic root and correlated well with histology (P < 0.0001). CONCLUSION: High-resolution MRI allows for accurate classification and quantification of atherosclerotic plaque components in the aortic root of mice.

McAteer MA, Schneider JE, Clarke K, Neubauer S, Channon KM, Choudhury RP. 2004. Quantification and 3D reconstruction of atherosclerotic plaque components in apolipoprotein E knockout mice using ex vivo high-resolution MRI. Arterioscler Thromb Vasc Biol, 24 (12), pp. 2384-2390. | Show Abstract | Read more

OBJECTIVE: To investigate the ability of high-resolution MRI to determine composition and microanatomy of atherosclerosis in mouse aortic root and brachiocephalic artery. METHODS AND RESULTS: Aortic root and brachiocephalic arteries of apolipoprotein E knockout (apoE-/-) mice fed Western diet for 10, 20, or 30 weeks were imaged ex vivo (11.7 T; 3D multiecho sequence; resolution 47x47x62.5 microm). Using semiautomated histogram-based methods, MRI accurately quantified lipid-rich/necrotic areas in the aortic root (r2=0.84; P<0.001) and brachiocephalic artery (r2=0.90; P<0.001) compared with histology. Similarly, cell-rich caps in aortic roots, quantified by MRI and histology, correlated closely (r2=0.74; P<0.001). Reconstruction of segmented brachiocephalic arteries in 3D provided unique insights into plaque microanatomy and enabled volumetric quantification of plaque and lipid-rich/necrotic core. Between 10 and 30 weeks, 3D measurement identified an 11.6-fold increase in plaque volume (versus 4.1-fold for 2D) and a 21.3-fold increase in plaque lipid-rich/necrotic core volume (versus 6.4-fold for 2D), indicating superior power of 3D quantification. CONCLUSIONS: Ex-vivo high-resolution 3D MRI accurately quantified lipid-rich/necrotic core and cell-rich cap areas in atherosclerotic lesions in apoE-/- mice. Reconstruction and volumetric quantification of segmented brachiocephalic arteries demonstrated greater sensitivity in detecting changes in plaque size and lipid composition over time than 2D analysis.

Schneider JE, Tyler DJ, ten Hove M, Sang AE, Cassidy PJ, Fischer A, Wallis J, Sebag-Montefiore LM, Watkins H, Isbrandt D et al. 2004. In vivo cardiac 1H-MRS in the mouse. Magn Reson Med, 52 (5), pp. 1029-1035. | Show Abstract | Read more

The mouse is the predominant animal model to study the effect of gene manipulations. Imaging techniques to define functional effects on the heart caused by genomic alterations are becoming increasingly routine in mice, yet methods for in vivo investigation of metabolic phenotypes in the mouse heart are lacking. In this work, cardiac 1H-MRS was developed and applied in mouse hearts in vivo using a single-voxel technique (PRESS). In normal C57Bl/6J mice, stability and reproducibility achieved by dedicated cardiac and respiratory gating was demonstrated by measuring amplitude and zero-order phase changes of the unsuppressed water signal. Various cardiac metabolites, such as creatine, taurine, carnitine, or intramyocardial lipids were successfully detected and quantified relative to the total water content in voxels as small as 2 microl, positioned in the interventricular septum. The method was applied to a murine model of guanidinoacetate N-methyltransferase (GAMT) deficiency, which is characterized by substantially decreased myocardial creatine levels. Creatine deficiency was confirmed noninvasively in myocardium of anesthetized GAMT-/- mice. This is the first study to report the application of cardiac 1H-MRS in mice in vivo.

Bamforth SD, Bragança J, Farthing CR, Schneider JE, Broadbent C, Michell AC, Clarke K, Neubauer S, Norris D, Brown NA et al. 2004. Cited2 controls left-right patterning and heart development through a Nodal-Pitx2c pathway. Nat Genet, 36 (11), pp. 1189-1196. | Show Abstract | Read more

Malformations of the septum, outflow tract and aortic arch are the most common congenital cardiovascular defects and occur in mice lacking Cited2, a transcriptional coactivator of TFAP2. Here we show that Cited2(-/-) mice also develop laterality defects, including right isomerism, abnormal cardiac looping and hyposplenia, which are suppressed on a mixed genetic background. Cited2(-/-) mice lack expression of the Nodal target genes Pitx2c, Nodal and Ebaf in the left lateral plate mesoderm, where they are required for establishing laterality and cardiovascular development. CITED2 and TFAP2 were detected at the Pitx2c promoter in embryonic hearts, and they activate Pitx2c transcription in transient transfection assays. We propose that an abnormal Nodal-Pitx2c pathway represents a unifying mechanism for the cardiovascular malformations observed in Cited2(-/-) mice, and that such malformations may be the sole manifestation of a laterality defect.

Dawson D, Lygate CA, Saunders J, Schneider JE, Ye X, Hulbert K, Noble JA, Neubauer S. 2004. Quantitative 3-dimensional echocardiography for accurate and rapid cardiac phenotype characterization in mice. Circulation, 110 (12), pp. 1632-1637. | Show Abstract | Read more

BACKGROUND: Insufficient techniques exist for rapid and reliable phenotype characterization of genetically manipulated mouse models of cardiac dysfunction. We developed a new, robust, 3-dimensional echocardiography (3D-echo) technique and hypothesized that this 3D-echo technique is as accurate as magnetic resonance imaging (MRI) and histology for assessment of left ventricular (LV) volume, ejection fraction, mass, and infarct size in normal and chronically infarcted mice. METHODS AND RESULTS: Using a high-frequency, 7/15-MHz, linear-array ultrasound transducer, we acquired ECG and respiratory-gated, 500-microm consecutive short-axis slices of the murine heart within 4 minutes. The short-axis movies were reassembled off-line in a 3D matrix by using the measured platform locations to position each slice in 3D. Epicardial and endocardial heart contours were manually traced, and a B-spline surface was fitted to the delineated image curves to reconstruct the heart volumes. Excellent correlations were obtained between 3D-echo and MRI for LV end-systolic volumes (r=0.99, P<0.0001), LV end-diastolic volumes (r=0.99, P<0.0001), ejection fraction (r=0.99, P<0.0001), LV mass (r=0.94, P<0.0019), and infarct size (r=0.98, P<0.0001). Also, excellent correlations were found between the 3D-echo-derived LV mass and necropsy LV mass in normal mice (r=0.99, P<0.0001), as well as for 3D-echo-derived infarct size and histologically determined infarct size (r=0.99, P<0.0001) in mice with chronic heart failure. Bland-Altman analysis showed excellent limits of agreement between techniques for all measured parameters. CONCLUSIONS: This new, fast, and highly reproducible 3D-echo technique should be of widespread applicability for high-throughput murine cardiac phenotyping studies.

Schneider JE, Bhattacharya S. 2004. Making the mouse embryo transparent: identifying developmental malformations using magnetic resonance imaging. Birth Defects Res C Embryo Today, 72 (3), pp. 241-249. | Show Abstract | Read more

Developmental malformations are a major cause of childhood mortality and are typically characterized by lesions that allow survival of the embryo through gestation. The genetics of developmental malformations are powerfully studied by using high-throughput, phenotype-driven screens (e.g., following zebrafish or mouse mutagenesis) or by genotype-driven studies using transgenic or knockout mice. With regard to either approach, the mouse is anatomically and phylogenetically closer to humans than any other genetically tractable model organism. This is particularly important in the cardiovascular and respiratory systems, which have unique mammalian features. The identification of murine models of developmental malformations is, however, hindered by the opacity of the late gestational mouse embryo. In this review, we describe recent advances in magnetic resonance imaging that make it possible to rapidly identify malformations in the developing mouse embryo with high efficiency.

Cassidy PJ, Schneider JE, Grieve SM, Lygate C, Neubauer S, Clarke K. 2004. Assessment of motion gating strategies for mouse magnetic resonance at high magnetic fields. J Magn Reson Imaging, 19 (2), pp. 229-237. | Show Abstract | Read more

PURPOSE: To assess the performance of motion gating strategies for mouse cardiac magnetic resonance (MR) at high magnetic fields by quantifying the levels of motion artifact observed in images and spectra in vivo. MATERIALS AND METHODS: MR imaging (MRI) of the heart, diaphragm, and liver; MR angiography of the aortic arch; and slice-selective 1H-spectroscopy of the heart were performed on anesthetized C57Bl/6 mice at 11.75 T. Gating signals were derived using a custom-built physiological motion gating device, and the gating strategies considered were no gating, cardiac gating, conventional gating (i.e., blanking during respiration), automatic gating, and user-defined gating. Both automatic and user-defined modes used cardiac and respiratory gating with steady-state maintenance during respiration. Gating performance was assessed by quantifying the levels of motion artifact observed in images and the degree of amplitude and phase stability in spectra. RESULTS: User-defined gating with steady-state maintenance during respiration gave the best performance for mouse cardiac imaging, angiography, and spectroscopy, with a threefold increase in signal intensity and a sixfold reduction in noise intensity compared to cardiac gating only. CONCLUSION: Physiological gating with steady-state maintenance during respiration is essential for mouse cardiac MR at high magnetic fields.

Schneider JE, Böse J, Bamforth SD, Gruber AD, Broadbent C, Clarke K, Neubauer S, Lengeling A, Bhattacharya S. 2004. Identification of cardiac malformations in mice lacking Ptdsr using a novel high-throughput magnetic resonance imaging technique. BMC Dev Biol, 4 pp. 16. | Show Abstract | Read more

BACKGROUND: Congenital heart defects are the leading non-infectious cause of death in children. Genetic studies in the mouse have been crucial to uncover new genes and signaling pathways associated with heart development and congenital heart disease. The identification of murine models of congenital cardiac malformations in high-throughput mutagenesis screens and in gene-targeted models is hindered by the opacity of the mouse embryo. RESULTS: We developed and optimized a novel method for high-throughput multi-embryo magnetic resonance imaging (MRI). Using this approach we identified cardiac malformations in phosphatidylserine receptor (Ptdsr) deficient embryos. These included ventricular septal defects, double-outlet right ventricle, and hypoplasia of the pulmonary artery and thymus. These results indicate that Ptdsr plays a key role in cardiac development. CONCLUSIONS: Our novel multi-embryo MRI technique enables high-throughput identification of murine models for human congenital cardiopulmonary malformations at high spatial resolution. The technique can be easily adapted for mouse mutagenesis screens and, thus provides an important new tool for identifying new mouse models for human congenital heart diseases.

Schneider JE, Cassidy PJ, Lygate C, Tyler DJ, Wiesmann F, Grieve SM, Hulbert K, Clarke K, Neubauer S. 2003. Fast, high-resolution in vivo cine magnetic resonance imaging in normal and failing mouse hearts on a vertical 11.7 T system. J Magn Reson Imaging, 18 (6), pp. 691-701. | Show Abstract | Read more

PURPOSE: To establish fast, high-resolution in vivo cine magnetic resonance imaging (cine-MRI) on a vertical 11.7-T MR system and to investigate the stability of normal and failing mouse hearts in the vertical position. MATERIALS AND METHODS: To optimize the method on a high-field system, various MR-related parameters, such as relaxation times and the need for respiratory gating, were quantitatively investigated. High-resolution cine-MRI was applied to normal mice and to a murine heart failure model. Cardiac functional parameters were compared to matched mice imaged previously on a horizontal MR system. RESULTS: A T(1) of 1.10 +/- 0.27 seconds and a T(2) of 18.5 +/- 3.9 msec were measured for murine myocardial tissue. A quantitative analysis also proved respiratory gating to be essential for obtaining artifact-free cine images in the vertical position at this field strength. Cardiac functional parameters of mice, obtained within one hour, agreed well with those from previous studies of mice in the horizontal position. CONCLUSION: This work shows that MR systems with a vertical bore design can be used to accurately measure cardiac function in both normal and chronically failing mouse hearts within one hour. The increased signal-to-noise ratio (SNR) due to the higher field strength could be exploited to obtain higher temporal and spatial resolution compared to previous studies that were performed on horizontal systems with lower field strengths.

Geng Y, Yu Q, Sicinska E, Das M, Schneider JE, Bhattacharya S, Rideout WM, Bronson RT, Gardner H, Sicinski P. 2003. Cyclin E ablation in the mouse. Cell, 114 (4), pp. 431-443. | Show Abstract | Read more

E type cyclins (E1 and E2) are believed to drive cell entry into the S phase. It is widely assumed that the two E type cyclins are critically required for proliferation of all cell types. Here, we demonstrate that E type cyclins are largely dispensable for mouse development. However, endoreplication of trophoblast giant cells and megakaryocytes is severely impaired in the absence of cyclin E. Cyclin E-deficient cells proliferate actively under conditions of continuous cell cycling but are unable to reenter the cell cycle from the quiescent G(0) state. Molecular analyses revealed that cells lacking cyclin E fail to normally incorporate MCM proteins into DNA replication origins during G(0)-->S progression. We also found that cyclin E-deficient cells are relatively resistant to oncogenic transformation. These findings define a molecular function for E type cyclins in cell cycle reentry and reveal a differential requirement for cyclin E in normal versus oncogenic proliferation.

Schneider JE, Bamforth SD, Farthing CR, Clarke K, Neubauer S, Bhattacharya S. 2003. Rapid identification and 3D reconstruction of complex cardiac malformations in transgenic mouse embryos using fast gradient echo sequence magnetic resonance imaging. J Mol Cell Cardiol, 35 (2), pp. 217-222. | Show Abstract | Read more

Developmental malformations of the heart in mouse embryos are commonly studied by histological sectioning. This is slow, labour intensive, and results in the loss of three-dimensional (3D) information. Magnetic resonance studies of embryos typically use spin-echo sequences, using prolonged acquisition times (>36 h) or perfusion with contrast agents to enhance resolution and contrast. This is technically difficult, and requires significant amounts of operator time. We imaged paraformaldehyde fixed embryos using a fast spoiled 3D gradient echo sequence with T(1)-weighting, in unattended overnight runs of less than 9 h. In wild-type embryos, we visualised normal cardiac structures, including cardiac chambers, the ventricular septum, primary and secondary atrial septa, valves, superior and inferior vena cava, aorta, pulmonary artery, and ductus arteriosus. In embryos lacking Cited2 (a transcriptional co-activator required for normal heart development), we identified cardiac malformations including atrial and ventricular septal defects, cono-truncal defects, and aortic arch malformations. We generated 3D reconstructions of normal and mutant hearts using contour identification and surface rendering computer software. The malformations were confirmed by histological sectioning. Our data indicate that fast gradient echo sequence magnetic resonance imaging can be used to rapidly and accurately identify complex cardiovascular malformations in transgenic and mutant mouse embryos.

Schneider JE, Bamforth SD, Grieve SM, Clarke K, Bhattacharya S, Neubauer S. 2003. High-resolution, high-throughput magnetic paragraph sign resonance imaging of mouse embryonic paragraph sign anatomy using a fast gradient-echo sequence. MAGMA, 16 (1), pp. 43-51. | Show Abstract | Read more

Embryonic development in normal and genetically modified mice is commonly analysed by histological sectioning. This procedure is time-consuming, prone to artefact, and results in the loss of three-dimensional (3D) information. Magnetic resonance imaging (MRI) of embryos has the potential of noninvasively acquiring a complete 3D data set. Published methods have used spin-echo techniques with inherently high signal-to-noise ratio (SNR); however, they required either perfusion of the embryo with a contrast agent, or prolonged acquisition times to improve contrast and resolution. Here, we show that a standard preparation (i.e. paraformaldehyde fixation) of 15.5 days post-coitum embryos followed by MRI using a fast gradient-echo sequence with T(1)-weighting achieves high resolution and high throughput for investigating mouse embryonic anatomy. 3D data sets were acquired in overnight experiments (<9 h) with an experimental resolution of approximately 25 microm(3). This spatial resolution is twofold higher than the values reported previously for comparable paraformaldehyde-fixed embryos, and it was obtained in less than a quarter of the time with sufficient SNR. Our approach combines speed, high resolution and contrast with a simple preparation technique and minimal operator time (<1 h). It allows rapid routine 3D characterisation of normal and abnormal mouse embryonic anatomy.

Schneider JE, Bamforth SD, Farthing CR, Clarke K, Neubauer S, Bhattacharya S. 2003. High-resolution imaging of normal anatomy, and neural and adrenal malformations in mouse embryos using magnetic resonance microscopy. J Anat, 202 (2), pp. 239-247. | Show Abstract | Read more

An efficient investigation of the effects of genetic or environmental manipulation on mouse development relies on the rapid and accurate screening of a substantial number of embryos for congenital malformations. Here we demonstrate that it is possible to examine normal organ development and identify malformations in mouse embryos by magnetic resonance microscopy in a substantially shorter time than by conventional histology. We imaged embryos in overnight runs of under 9 h, with an operator time of less than 1 h. In normal embryos we visualized the brain, spinal cord, ganglia, eyes, inner ear, pituitary, thyroid, thymus, trachea, bronchi, lungs, heart, kidneys, gonads, adrenals, oesophagus, stomach, intestines, spleen, liver and pancreas. Examination of the brain in embryos lacking the transcriptional coactivator Cited2 showed cerebellar and midbrain roof agenesis, in addition to exencephaly. In these embryos we were also able to detect agenesis of the adrenal gland. We confirmed all malformations by histological sectioning. Thus magnetic resonance microscopy can be used to rapidly identify developmental and organ malformations in mutant mouse embryos generated by transgenic techniques, in high-throughput mutagenesis screens, or in screens to identify teratogenic compounds and environmental factors contributing to developmental malformations.

Grieve SM, Choudhury RP, Schneider JE, Cassidy PJ, Neubauer S, Clarke K. 2002. Magnetic resonance angiography of mouse aorta at 12 tesla ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 22 (5), pp. A12-A12.

Grieve SM, Priestman DA, Schneider JE, Platt FM, Dwek R, Neubauer S, Clarke K. 2002. An MRI technique for the quantification of atherosclerotic lesions in the ApoE knockout mouse ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 22 (5), pp. A48-A48.

Schneider J, Fekete E, Weisser A, Neubauer S, von Kienlin M. 2000. Reduced (1)H-NMR visibility of creatine in isolated rat hearts. Magn Reson Med, 43 (4), pp. 497-502. | Show Abstract | Read more

The aim of this study was to measure the concentration of creatine in Langendorff perfused rat hearts, both by quantitative (1)H-MRS and by high-pressure liquid chromatography (HPLC). First, the relaxation times and other parameters affecting absolute quantification by MRS were determined. At 11.75 T, the relaxation times of myocardial creatine were T(1) = 1.1+/-0.29 sec (mean +/- SD, n = 5) and T(2) = 56.4+/-6.2 ms (n = 9). In phantom experiments the MRS measurements gave accurate values for the known relative concentrations of the detected substances. In glucose-perfused rat hearts, the creatine concentration measured by HPLC was 14.2+/-1.9 mmol/kg wet weight (n = 8), in good agreement with literature values. The (1)H-MRS measurements, however, resulted in creatine concentrations of only approximately 60% of this value. The application of CHESS-pulses for water suppression led to a further 30% reduction of the creatine MRS signal. These results indicate a reduced (1)H-NMR visibility of creatine in the myocardium, which suggests a compartmentation of myocardial creatine into various pools.

Casero R, Siedlecka U, Jones ES, Gruscheski L, Gibb M, Schneider JE, Kohl P, Grau V. 2017. Transformation diffusion reconstruction of three-dimensional histology volumes from two-dimensional image stacks. Med Image Anal, 38 pp. 184-204. | Show Abstract | Read more

Traditional histology is the gold standard for tissue studies, but it is intrinsically reliant on two-dimensional (2D) images. Study of volumetric tissue samples such as whole hearts produces a stack of misaligned and distorted 2D images that need to be reconstructed to recover a congruent volume with the original sample's shape. In this paper, we develop a mathematical framework called Transformation Diffusion (TD) for stack alignment refinement as a solution to the heat diffusion equation. This general framework does not require contour segmentation, is independent of the registration method used, and is trivially parallelizable. After the first stack sweep, we also replace registration operations by operations in the space of transformations, several orders of magnitude faster and less memory-consuming. Implementing TD with operations in the space of transformations produces our Transformation Diffusion Reconstruction (TDR) algorithm, applicable to general transformations that are closed under inversion and composition. In particular, we provide formulas for translation and affine transformations. We also propose an Approximated TDR (ATDR) algorithm that extends the same principles to tensor-product B-spline transformations. Using TDR and ATDR, we reconstruct a full mouse heart at pixel size 0.92µm×0.92µm, cut 10µm thick, spaced 20µm (84G). Our algorithms employ only local information from transformations between neighboring slices, but the TD framework allows theoretical analysis of the refinement as applying a global Gaussian low-pass filter to the unknown stack misalignments. We also show that reconstruction without an external reference produces large shape artifacts in a cardiac specimen while still optimizing slice-to-slice alignment. To overcome this problem, we use a pre-cutting blockface imaging process previously developed by our group that takes advantage of Brewster's angle and a polarizer to capture the outline of only the topmost layer of wax in the block containing embedded tissue for histological sectioning.

Bates J, Teh I, McClymont D, Kohl P, Schneider JE, Grau V. 2017. Monte Carlo Simulations of Diffusion Weighted MRI in Myocardium: Validation and Sensitivity Analysis. IEEE Trans Med Imaging, 36 (6), pp. 1316-1325. | Show Abstract | Read more

A model of cardiac microstructure and diffusion MRI is presented, and compared with experimental data from ex vivo rat hearts. The model includes a simplified representation of individual cells, with physiologically correct cell size and orientation, as well as intra- to extracellular volume ratio. Diffusion MRI is simulated using a Monte Carlo model and realistic MRI sequences. The results show good correspondence between the simulated and experimental MRI signals. Similar patterns are observed in the eigenvalues of the diffusion tensor, the mean diffusivity (MD), and the fractional anisotropy (FA). A sensitivity analysis shows that the diffusivity is the dominant influence on all three eigenvalues of the diffusion tensor, the MD, and the FA. The area and aspect ratio of the cell cross-section affect the secondary and tertiary eigenvalues, and hence the FA. Within biological norms, the cell length, volume fraction of cells, and rate of change of helix angle play a relatively small role in influencing tissue diffusion. Results suggest that the model could be used to improve understanding of the relationship between cardiac microstructure and diffusion MRI measurements, as well as in testing and refinement of cardiac diffusion MRI protocols.

Constantinides C, Maguire ML, Stork L, Swider E, Srinivas M, Carr CA, Schneider JE. 2017. Temporal accumulation and localization of isoflurane in the C57BL/6 mouse and assessment of its potential contamination in (19) F MRI with perfluoro-crown-ether-labeled cardiac progenitor cells at 9.4 Tesla. J Magn Reson Imaging, 45 (6), pp. 1659-1667. | Show Abstract | Read more

PURPOSE: To assess the uptake, accumulation, temporal stability, and spatial localization of isoflurane (ISO) in the C57BL/6 mouse, and to identify its potential interference with the detection of labeled cardiac progenitor cells using (19) F MRI/MR spectroscopy (MRS). MATERIALS AND METHODS: Objectives are demonstrated using (a) in vitro ISO tests, (b) in vivo temporal accumulation/spatial localization C57BL/6 studies (n = 3), and (c) through injections of perfluoro-crown-ether (PFCE) labeled cardiac progenitor cells into femoral muscle areas of the murine hindlimb post-mortem (n = 1) using (1) H/(19) F MRI/MRS at 9.4 Tesla. Data were acquired using double-gated spoiled gradient echo images and pulse-acquire spectra. For the in vivo study, the temporal stability of ISO resonances was quantified using coefficient of variability (CV) (5 min) estimates. RESULTS: Two ISO resonances were observed in vivo that correspond to the -CF3 and -OCHF2 moieties. CV values ranged between 3.2 and 6.4% (-CF3 ) and 6.4 and 11.2% (-OCHF2 ). Reductions of the ISO dose (2.0 to 1.7%) at 80 min postinduction had insignificant effects on ISO signals (P = 0.23; P = 0.71). PFCE-labeled cells exhibited a resonance at -16.25 ppm in vitro that did not overlap with the ISO resonances, a finding that is confirmed with MRS post-mortem using injected, labeled cells. Based on (19) F MRI, similar in vivo/post-mortem ISO compartmentalization was also confirmed in peripheral and thoracic skeletal muscles. CONCLUSION: Significant ISO accumulation was observed by (19) F MRS in vivo with temporally stable signals over 90 min postinduction. ISO effects on PFCE labels are anticipated to be minimal but may be more prominent for perfluoropolyether or perfluorooctyl bromide labels. LEVEL OF EVIDENCE: 1 Technical Efficacy: Stage 1 J. MAGN. RESON. IMAGING 2017;45:1659-1667.

McClymont D, Teh I, Carruth E, Omens J, McCulloch A, Whittington HJ, Kohl P, Grau V, Schneider JE. 2016. Evaluation of non-Gaussian diffusion in cardiac MRI. Magn Reson Med, | Show Abstract | Read more

PURPOSE: The diffusion tensor model assumes Gaussian diffusion and is widely applied in cardiac diffusion MRI. However, diffusion in biological tissue deviates from a Gaussian profile as a result of hindrance and restriction from cell and tissue microstructure, and may be quantified better by non-Gaussian modeling. The aim of this study was to investigate non-Gaussian diffusion in healthy and hypertrophic hearts. METHODS: Thirteen rat hearts (five healthy, four sham, four hypertrophic) were imaged ex vivo. Diffusion-weighted images were acquired at b-values up to 10,000 s/mm(2) . Models of diffusion were fit to the data and ranked based on the Akaike information criterion. RESULTS: The diffusion tensor was ranked best at b-values up to 2000 s/mm(2) but reflected the signal poorly in the high b-value regime, in which the best model was a non-Gaussian "beta distribution" model. Although there was considerable overlap in apparent diffusivities between the healthy, sham, and hypertrophic hearts, diffusion kurtosis and skewness in the hypertrophic hearts were more than 20% higher in the sheetlet and sheetlet-normal directions. CONCLUSION: Non-Gaussian diffusion models have a higher sensitivity for the detection of hypertrophy compared with the Gaussian model. In particular, diffusion kurtosis may serve as a useful biomarker for characterization of disease and remodeling in the heart. Magn Reson Med, 2016. © 2016 The Authors Magnetic Resonance in Medicine published by Wiley Periodicals, Inc. on behalf of International Society for Magnetic Resonance in Medicine. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Teh I, McClymont D, Burton RA, Maguire ML, Whittington HJ, Lygate CA, Kohl P, Schneider JE. 2016. Resolving Fine Cardiac Structures in Rats with High-Resolution Diffusion Tensor Imaging. Sci Rep, 6 (1), pp. 30573. | Show Abstract | Read more

Cardiac architecture is fundamental to cardiac function and can be assessed non-invasively with diffusion tensor imaging (DTI). Here, we aimed to overcome technical challenges in ex vivo DTI in order to extract fine anatomical details and to provide novel insights in the 3D structure of the heart. An integrated set of methods was implemented in ex vivo rat hearts, including dynamic receiver gain adjustment, gradient system scaling calibration, prospective adjustment of diffusion gradients, and interleaving of diffusion-weighted and non-diffusion-weighted scans. Together, these methods enhanced SNR and spatial resolution, minimised orientation bias in diffusion-weighting, and reduced temperature variation, enabling detection of tissue structures such as cell alignment in atria, valves and vessels at an unprecedented level of detail. Improved confidence in eigenvector reproducibility enabled tracking of myolaminar structures as a basis for segmentation of functional groups of cardiomyocytes. Ex vivo DTI facilitates acquisition of high quality structural data that complements readily available in vivo cardiac functional and anatomical MRI. The improvements presented here will facilitate next generation virtual models integrating micro-structural and electro-mechanical properties of the heart.

Bueno-Orovio A, Teh I, Schneider JE, Burrage K, Grau V. 2016. Anomalous Diffusion in Cardiac Tissue as an Index of Myocardial Microstructure. IEEE Trans Med Imaging, 35 (9), pp. 2200-2207. | Show Abstract | Read more

Diffusion in biological tissues is known to be hindered by the structural complexity of the underlying medium. In the heart, improved characterisation on how this complexity influences acquired diffusion weighted signals is key to advancing our interpretation of diffusion magnetic resonance imaging, as well as to propose novel biomarkers to further characterise myocardial microstructure. In this work, we propose stretched Mittag-Leffler signal decay models for the quantification of the anomalous decay observed in acquired diffusion weighted signals. Our results, analysed in ex vivo healthy, fixed rat ventricles, indicate that such a representation suffices to capture the anomalous signal decay observed in the myocardial syncytium. The subdiffusive order of signal decay is shown to encode independent information to that encapsulated by standard diffusion tensor metrics, and thus may provide additional information on tissue microstructure. Moreover, subdiffusion gradients are shown to be indicative of the total structural heterogeneity spanning the left ventricular wall, which includes progressive myolaminae branching and spatially varying densities of perimysial collagen, microvasculature and adipose tissue. The proposed approach may therefore have important implications for the characterisation of tissue microstructure, both in cardiac and other tissue types.

Dall'Armellina E, Jung BA, Lygate CA, Neubauer S, Markl M, Schneider JE. 2012. Improved method for quantification of regional cardiac function in mice using phase-contrast MRI Magnetic Resonance in Medicine, 67 (2), pp. 541-551. | Show Abstract | Read more

Phase-contrast magnetic resonance imaging is a technique that allows for characterization of regional cardiac function and for measuring transmural myocardial velocities in human hearts with high temporal and spatial resolution. The application of this technique (also known as tissue phase mapping) to murine hearts has been very limited so far. The aim of our study was to implement and to optimize tissue phase mapping for a comprehensive assessment of murine transmural wall motion. Baseline values for regional motion patterns in mouse hearts, based on the clinically used American Heart Association's 17-segment model, were established, and a detailed motion analysis of mouse heart for the entire cardiac cycle (including epicardial and endocardial motion patterns) is provided. Black-blood contrast was found to be essential to obtain reproducible velocity encoding. Tissue phase mapping of the mouse heart permits the detailed assessment of regional myocardial velocities. While a proof-of-principle application in a murine ischemia-reperfusion model was performed, future studies are warranted to assess its potential for the investigation of systolic and diastolic functions in genetically and surgically manipulated mouse models of human heart disease. Magn Reson Med, 2012. © 2011 Wiley Periodicals, Inc.

Wech T, Lemke A, Medway D, Stork LA, Lygate CA, Neubauer S, Köstler H, Schneider JE. 2011. Accelerating cine-MR imaging in mouse hearts using compressed sensing. J Magn Reson Imaging, 34 (5), pp. 1072-1079. | Show Abstract | Read more

PURPOSE: To combine global cardiac function imaging with compressed sensing (CS) in order to reduce scan time and to validate this technique in normal mouse hearts and in a murine model of chronic myocardial infarction. MATERIALS AND METHODS: To determine the maximally achievable acceleration factor, fully acquired cine data, obtained in sham and chronically infarcted (MI) mouse hearts were 2-4-fold undersampled retrospectively, followed by CS reconstruction and blinded image segmentation. Subsequently, dedicated CS sampling schemes were implemented at a preclinical 9.4 T magnetic resonance imaging (MRI) system, and 2- and 3-fold undersampled cine data were acquired in normal mouse hearts with high temporal and spatial resolution. RESULTS: The retrospective analysis demonstrated that an undersampling factor of three is feasible without impairing accuracy of cardiac functional parameters. Dedicated CS sampling schemes applied prospectively to normal mouse hearts yielded comparable left-ventricular functional parameters, and intra- and interobserver variability between fully and 3-fold undersampled data. CONCLUSION: This study introduces and validates an alternative means to speed up experimental cine-MRI without the need for expensive hardware.

Hales PW, Burton RA, Bollensdorff C, Mason F, Bishop M, Gavaghan D, Kohl P, Schneider JE. 2011. Progressive changes in T₁, T₂ and left-ventricular histo-architecture in the fixed and embedded rat heart. NMR Biomed, 24 (7), pp. 836-843. | Show Abstract | Read more

Chemical tissue fixation, followed by embedding in either agarose or Fomblin, is common practice in time-intensive MRI studies of ex vivo biological samples, and is required to prevent tissue autolysis and sample motion. However, the combined effect of fixation and sample embedding may alter tissue structure and MRI properties. We investigated the progressive changes in T(1) and T(2) relaxation times, and the arrangement of locally prevailing cardiomyocyte orientation determined using diffusion tensor imaging, in embedded ex vivo rat hearts fixed using Karnovsky's solution (glutaraldehyde-formaldehyde mix). Three embedding media were investigated: (i) standard agarose (n = 3 hearts); (ii) Fomblin (n = 4 hearts); and (iii) iso-osmotic agarose (n = 3 hearts); in the latter, the osmolarity of the fixative and embedding medium was adjusted to 300 mOsm to match more closely that of native tissue. The T(1) relaxation time in the myocardium showed a pronounced decrease over a 48-h period following embedding in Fomblin (-11.3 ± 6.2%; mean ± standard deviation), but was stable in standard agarose- and iso-osmotic agarose-embedded hearts. The mean myocardial T(2) relaxation time increased in all embedded hearts: by 35.1 ± 14.7% with standard agarose embedding, 13.1 ± 5.6% with Fomblin and 13.3 ± 1.4% with iso-osmotic agarose. Deviation in the orientation of the primary eigenvector of the diffusion tensor occurred in all hearts (mean angular changes of 6.6°, 3.2° and 1.9° per voxel after 48 h in agarose-, Fomblin- and iso-osmotic agarose-embedded hearts, respectively), indicative of progressive structural changes in myocardial histo-architecture, in spite of previous exposure to fast-acting tissue fixation. Our results suggest that progressive structural changes occur in chemically fixed myocardium, and that the extent of these changes is modulated by the embedding medium, and by osmotic gradients between the fixative in the tissue and the surrounding medium.

Rial B, Robson MD, Neubauer S, Schneider JE. 2011. Rapid quantification of myocardial lipid content in humans using single breath-hold 1H MRS at 3 Tesla. Magn Reson Med, 66 (3), pp. 619-624. | Show Abstract | Read more

A rapid, proton magnetic resonance spectroscopy method to evaluate human myocardial lipid levels in a single breath-hold at 3 T using a commercial whole-body system is presented. During a 10 s breath-hold, water unsuppressed and suppressed spectra were acquired by two phased array coils using a short-echo time spectroscopic stimulated echo (STEAM) sequence electrocardiogram-triggered to mid-diastole. Lipid-to-water ratios were obtained in the septum of 15 healthy volunteers, (0.46 ± 0.19)%. These results agreed well with ratios obtained from averaged spectra acquired in seven multiple breath-holds, (0.45 ± 0.20)%, providing increased signal-to-noise ratio but requiring longer acquisition times. Excellent correlation was found between the two methods (r=0.94, P<0.05). Reproducibility of (1)H MRS for measuring myocardial lipid levels in a short breath-hold was acceptable in five repeated measurements within the same subject (coefficient of variation=19%). Thus, single breath-hold proton spectroscopy allows reliable and quick quantification of myocardial lipids at 3 T.

Schneider JE, Lanz T, Barnes H, Stork LA, Bohl S, Lygate CA, Ordidge RJ, Neubauer S. 2011. Accelerated cardiac magnetic resonance imaging in the mouse using an eight-channel array at 9.4 Tesla. Magn Reson Med, 65 (1), pp. 60-70. | Show Abstract | Read more

MRI has become an important tool to noninvasively assess global and regional cardiac function, infarct size, or myocardial blood flow in surgically or genetically modified mouse models of human heart disease. Constraints on scan time due to sensitivity to general anesthesia in hemodynamically compromised mice frequently limit the number of parameters available in one imaging session. Parallel imaging techniques to reduce acquisition times require coil arrays, which are technically challenging to design at ultrahigh magnetic field strengths. This work validates the use of an eight-channel volume phased-array coil for cardiac MRI in mice at 9.4 T. Two- and three-dimensional sequences were combined with parallel imaging techniques and used to quantify global cardiac function, T(1)-relaxation times and infarct sizes. Furthermore, the rapid acquisition of functional cine-data allowed for the first time in mice measurement of left-ventricular peak filling and ejection rates under intravenous infusion of dobutamine. The results demonstrate that a threefold accelerated data acquisition is generally feasible without compromising the accuracy of the results. This strategy may eventually pave the way for routine, multiparametric phenotyping of mouse hearts in vivo within one imaging session of tolerable duration.

Schneider JE, Lanz T, Barnes H, Medway D, Stork LA, Lygate CA, Smart S, Griswold MA, Neubauer S. 2008. Ultra-fast and accurate assessment of cardiac function in rats using accelerated MRI at 9.4 Tesla. Magn Reson Med, 59 (3), pp. 636-641. | Show Abstract | Read more

MRI can accurately and reproducibly assess cardiac function in rodents but requires relatively long imaging times. Therefore, parallel imaging techniques using a 4-element RF-coil array and MR sequences for cardiac MRI in rats were implemented at ultra-high magnetic fields (9.4 Tesla [T]). The hypothesis that these developments would result in a major reduction in imaging time without loss of accuracy was tested on female Wistar rats under isoflurane anesthesia. High-resolution, contiguous short-axis slices (thickness 1.5 mm) were acquired covering the entire heart. Two interleaved data sets (i) with the volume coil (eight averages) and (ii) with the four-element coil array (one average) were obtained. In addition, two-, three-, and fourfold accelerated data sets were generated through postprocessing of the coil array data, followed by a TGRAPPA reconstruction, resulting in five data sets per rat (in-plane voxel size 100 x 100 microm). Using a single blinded operator, excellent agreement was obtained between volume coil (acquisition time: 88 min) and the fourfold accelerated (<3 min) data sets (e.g., LV mass 436 +/- 21 mg vs 433 +/- 19 mg; ejection fraction 74 +/- 5% vs 75 +/- 4%). This finding demonstrates that it is possible to complete a rat cine-MRI study under 3 min with low variability and without losing temporal or spatial resolution, making high throughput screening programs feasible.

Schneider JE, Tyler DJ, ten Hove M, Sang AE, Cassidy PJ, Fischer A, Wallis J, Sebag-Montefiore LM, Watkins H, Isbrandt D et al. 2004. In vivo cardiac 1H-MRS in the mouse. Magn Reson Med, 52 (5), pp. 1029-1035. | Show Abstract | Read more

The mouse is the predominant animal model to study the effect of gene manipulations. Imaging techniques to define functional effects on the heart caused by genomic alterations are becoming increasingly routine in mice, yet methods for in vivo investigation of metabolic phenotypes in the mouse heart are lacking. In this work, cardiac 1H-MRS was developed and applied in mouse hearts in vivo using a single-voxel technique (PRESS). In normal C57Bl/6J mice, stability and reproducibility achieved by dedicated cardiac and respiratory gating was demonstrated by measuring amplitude and zero-order phase changes of the unsuppressed water signal. Various cardiac metabolites, such as creatine, taurine, carnitine, or intramyocardial lipids were successfully detected and quantified relative to the total water content in voxels as small as 2 microl, positioned in the interventricular septum. The method was applied to a murine model of guanidinoacetate N-methyltransferase (GAMT) deficiency, which is characterized by substantially decreased myocardial creatine levels. Creatine deficiency was confirmed noninvasively in myocardium of anesthetized GAMT-/- mice. This is the first study to report the application of cardiac 1H-MRS in mice in vivo.

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