A multi‐Gaussian model for apparent diffusion coefficient histogram analysis of Wilms’ tumour subtype and response to chemotherapy

Wilms’ tumours (WTs) are large heterogeneous tumours, which typically consist of a mixture of histological cell types, together with regions of chemotherapy‐induced regressive change and necrosis. The predominant cell type in a WT is assessed histologically following nephrectomy, and used to assess the tumour subtype and potential risk. The purpose of this study was to develop a mathematical model to identify subregions within WTs with distinct cellular environments in vivo, determined using apparent diffusion coefficient (ADC) values from diffusion‐weighted imaging (DWI). We recorded the WT subtype from the histopathology of 32 tumours resected in patients who received DWI prior to surgery after pre‐operative chemotherapy had been administered. In 23 of these tumours, DWI data were also available prior to chemotherapy. Histograms of ADC values were analysed using a multi‐Gaussian model fitting procedure, which identified ‘subpopulations’ with distinct cellular environments within the tumour volume. The mean and lower quartile ADC values of the predominant viable tissue subpopulation (ADC_1MEAN, ADC_1LQ), together with the same parameters from the entire tumour volume (ADC_0MEAN, ADC_0LQ), were tested as predictors of WT subtype. ADC_1LQ from the multi‐Gaussian model was the most effective parameter for the stratification of WT subtype, with significantly lower values observed in high‐risk blastemal‐type WTs compared with intermediate‐risk stromal, regressive and mixed‐type WTs (p < 0.05). No significant difference in ADC_1LQ was found between blastemal‐type and intermediate‐risk epithelial‐type WTs. The predominant viable tissue subpopulation in every stromal‐type WT underwent a positive shift in ADC_1MEAN after chemotherapy. Our results suggest that our multi‐Gaussian model is a useful tool for differentiating distinct cellular regions within WTs, which helps to identify the predominant histological cell type in the tumour in vivo. This shows potential for improving the risk‐based stratification of patients at an early stage, and for guiding biopsies to target the most malignant part of the tumour. Copyright © 2015 John Wiley & Sons, Ltd.     

Assessment of non‐Gaussian diffusion with singly and doubly stretched biexponential models of diffusion‐weighted MRI (DWI) signal attenuation in prostate tissue

Non‐Gaussian diffusion dynamics was investigated in the two distinct water populations identified by a biexponential model of diffusion in prostate tissue. Diffusion‐weighted MRI (DWI) signal attenuation was measured ex vivo in two formalin‐fixed prostates at 9.4 T with diffusion times Δ = 10, 20 and 40 ms, and b values in the range 0.017–8.2 ms/µm². A conventional biexponential model was compared with models in which either the lower diffusivity component or both of the components of the biexponential were stretched. Models were compared using Akaike's Information Criterion (AIC) and a leave‐one‐out (LOO) test of model prediction accuracy. The doubly stretched (SS) model had the highest LOO prediction accuracy and lowest AIC (highest information content) in the majority of voxels at Δ = 10 and 20 ms. The lower diffusivity stretching factor (α₂) of the SS model was consistently lower (range ~0.3–0.9) than the higher diffusivity stretching factor (α₁, range ~0.7–1.1), indicating a high degree of diffusion heterogeneity in the lower diffusivity environment, and nearly Gaussian diffusion in the higher diffusivity environment. Stretched biexponential models demonstrate that, in prostate tissue, the two distinct water populations identified by the simple biexponential model individually exhibit non‐Gaussian diffusion dynamics. Copyright © 2015 John Wiley & Sons, Ltd.     

A quantitative comparison of metabolite signals as detected by in vivo MRS with ex vivo 1H HR‐MAS for childhood brain tumours

¹H MRS provides a powerful method for investigating tumour metabolism by allowing the measurement of metabolites in vivo. Recently, the technique of ¹H high‐resolution magic angle spinning (HR‐MAS) has been shown to produce high‐quality data, allowing the accurate measurement of many metabolites present in unprocessed biopsy tissue. The purpose of this study was to evaluate the agreement between the techniques of in vivo MRS and ex vivo HR‐MAS for investigating childhood brain tumours. Short‐TE (30 ms), single‐voxel, in vivo MRS was performed on 16 paediatric patients with brain tumours at 1.5 T. A frozen biopsy sample was available for each patient. HR‐MAS was performed on the biopsy samples, and metabolite quantities were determined from the MRS and HR‐MAS data using the LCModel? and TARQUIN algorithms, respectively. Linear regression was performed on the metabolite quantities to asses the agreement between MRS and HR‐MAS. Eight of the 12 metabolite quantities were found to correlate significantly (P < 0.05). The four worst correlating metabolites were aspartate, scyllo‐inositol, glycerophosphocholine and N‐acetylaspartate, and, except for glycerophosphocholine, this error was reflected in their higher Cramer–Rao lower bounds (CRLBs), suggesting that low signal‐to‐noise was the greatest source of error for these metabolites. Glycerophosphocholine had a lower CRLB implying that interference with phosphocholine and choline was the most significant source of error. The generally good agreement observed between the two techniques suggests that both MRS and HR‐MAS can be used to reliably estimate metabolite quantities in brain tumour tissue and that tumour heterogeneity and metabolite degradation do not have an important effect on the HR‐MAS metabolite profile for the tumours investigated. HR‐MAS can be used to improve the analysis and understanding of MRS data. Copyright © 2009 John Wiley & Sons, Ltd.     

Assessment of early docetaxel response in an experimental model of human breast cancer using DCE‐MRI,ex vivo HR MAS,and in vivo 1H MRS

The purpose of this study was to evaluate the use of dynamic contrast‐enhanced (DCE) MRI, in vivo ¹H MRS and ex vivo high resolution magic angle spinning (HR MAS) MRS of tissue samples as methods to detect early treatment effects of docetaxel in a breast cancer xenograft model (MCF‐7) in mice. MCF‐7 cells were implanted subcutaneously in athymic mice and treated with docetaxel (20, 30, and 40 mg/kg) or saline six weeks later. DCE‐MRI and in vivo ¹H MRS were performed on a 7 T MR system three days after treatment. The dynamic images were used as input for a two‐compartment model, yielding the vascular parameters K^trans and v_e. HR MAS MRS, histology, and immunohistochemical staining for proliferation (Ki‐67), apoptosis (M30 cytodeath), and vascular/endothelial cells (CD31) were performed on excised tumor tissue. Both in vivo spectra and HR MAS spectra were used as input for multivariate analysis (principal component analysis (PCA) and partial least squares regression analysis (PLS)) to compare controls to treated tumors. Tumor growth was suppressed in docetaxel‐treated mice compared to the controls. The anti‐tumor effect led to an increase in K^trans and v_e values in all the treated groups. Furthermore, in vivo MRS and HR MAS MRS revealed a significant decrease in choline metabolite levels for the treated groups, in accordance with reduced proliferative index as seen on Ki‐67 stained sections. In this study DCE‐MRI, in vivo MRS and ex vivo HR MAS MRS have been used to demonstrate that docetaxel treatment of a human breast cancer xenograft model results in changes in the vascular dynamics and metabolic profile of the tumors. This indicates that these MR methods could be used to monitor intra‐tumoral treatment effects. Copyright © 2009 John Wiley & Sons, Ltd.     

In vivo measurement of apparent diffusion coefficients of hyperpolarized 13C‐labeled metabolites

The combination of hyperpolarized MRS with diffusion weighting (dw) allows for determination of the apparent diffusion coefficient (ADC), which is indicative of the intra‐ or extracellular localization of the metabolite. Here, a slice‐selective pulsed‐gradient spin echo sequence was implemented to acquire a series of dw spectra from rat muscle in vivo to determine the ADCs of multiple metabolites after a single injection of hyperpolarized [1‐¹³C]pyruvate. An optimal control optimized universal‐rotation pulse was used for refocusing to minimize signal loss caused by B₁ imperfections. Non‐dw spectra were acquired interleaved with the dw spectra and these were used to correct for signal decay during the acquisition as a result of T₁ decay, pulse imperfections, flow etc. The data showed that the ADC values for [1‐¹³C]lactate (0.4–0.7 µm²/ms) and [1‐¹³C]alanine (0.4–0.9 µm²/ms) were about a factor of two lower than the ADC of [1‐¹³C]pyruvate (1.1–1.5 µm²/ms). This indicates a more restricted diffusion space for the former two metabolites consistent with lactate and alanine being intracellular. The higher ADC for pyruvate (similar to the proton ADC) reflected that the injected substance was not confined inside the muscle cells but also present extracellular. Copyright © 2014 John Wiley & Sons, Ltd.     

PARP inhibitor rucaparib induces changes in NAD levels in cells and liver tissues as assessed by MRS

Poly(adenosine diphosphate ribose) polymerases (PARPs) are multifunctional proteins which play a role in many cellular processes. Namely, PARP1 and PARP2 have been shown to be involved in DNA repair, and therefore are valid targets in cancer treatment with PARP inhibitors, such as rucaparib, currently in clinical trials. Proton magnetic resonance spectroscopy (¹H‐MRS) was used to study the impact of rucaparib in vitro and ex vivo in liver tissue from mice, via quantitative analysis of nicotinamide adenosine diphosphate (NAD⁺) spectra, to assess the potential of MRS as a biomarker of the PARP inhibitor response. SW620 (colorectal) and A2780 (ovarian) cancer cell lines, and PARP1 wild‐type (WT) and PARP1 knock‐out (KO) mice, were treated with rucaparib, temozolomide (methylating agent) or a combination of both drugs. ¹H‐MRS spectra were obtained from perchloric acid extracts of tumour cells and mouse liver. Both cell lines showed an increase in NAD⁺ levels following PARP inhibitor treatment in comparison with temozolomide treatment. Liver extracts from PARP1 WT mice showed a significant increase in NAD⁺ levels after rucaparib treatment compared with untreated mouse liver, and a significant decrease in NAD⁺ levels in the temozolomide‐treated group. The combination of rucaparib and temozolomide did not prevent the NAD⁺ depletion caused by temozolomide treatment. The ¹H‐MRS results show that NAD⁺ levels can be used as a biomarker of PARP inhibitor and methylating agent treatments, and suggest that in vivo measurement of NAD⁺ would be valuable.     

Sequence design and evaluation of the reproducibility of water‐selective diffusion‐weighted imaging of the breast at 3 T

Diffusion measurements derived from breast MRI can be adversely affected by unwanted signals from abundant fatty tissues if they are not suppressed adequately. To minimize this undesired contribution, we designed and optimized a water‐selective diffusion‐weighted imaging (DWI) sequence, which relies on spectrally selective excitation on the water resonance, obviating the need for fat suppression. As this method is more complex than standard DWI methods, we also report a test–retest study to evaluate its reproducibility. In this study, a spectrally selective Gaussian pulse on water resonance was combined with a pair of slice‐selective adiabatic refocusing pulses for water‐only DWI. Field map‐based shimming and manual determination of the center frequency were used for water selection. The selectivity of the excitation pulse was optimized by a spectrally selective spectroscopy sequence based on the same principles. A test–retest study of 10 volunteers in two separate visits was used to evaluate its reproducibility. Our results from all subjects showed high‐quality diffusion‐weighted images of the breast without fat contamination. Mean apparent diffusion coefficients for b = 0, 600 s/mm² and b = 50, 600 s/mm² all showed good reproducibility, as 95% confidence intervals of the apparent diffusion coefficients were 4 × 10^–5 mm²/s and 5 × 10^–5 mm²/s and repeatability values were 1.09 × 10^–4 and 1.31 × 10^–4, respectively. In conclusion, water‐selective DWI is a feasible alternative to standard methods of DWI based on fat suppression. The added complexity of the method does not compromise the reproducibility of diffusion measurements in the breast. Copyright © 2014 John Wiley & Sons, Ltd.     

Considerations for measuring the fractional anisotropy of metabolites with diffusion tensor spectroscopy

Diffusion tensor spectroscopy of metabolites in brain is challenging because of their lower diffusivity (i.e. less signal attenuation for a given b value) and much poorer signal‐to‐noise ratio relative to water. Although diffusion tensor acquisition protocols have been studied in detail for water, they have not been evaluated systematically for the measurement of the fractional anisotropy of metabolites such as N‐acetylaspartate, creatine and choline in the white and gray matter of human brain. Diffusion tensor spectroscopy was performed in vivo with variable maximal b values (1815 or 5018 s/mm²). Experiments were also performed on simulated spectra and isotropic alcohol phantoms of various diffusivities, ranging from approximately 0.54 × 10^?3 to 0.13 × 10^?3 mm²/s, to assess the sensitivity of diffusion tensor spectroscopic parameters to low diffusivity, noise and b value. The low maximum b value of 1815 s/mm² yielded elevated fractional anisotropy (0.53–0.60) of N‐acetylaspartate in cortical gray matter relative to the more isotropic value (0.25–0.30) obtained with a higher b value of 5018 s/mm²; in contrast, the fractional anisotropy of white matter was consistently anisotropic with the different maximal b values (i.e. 0.43–0.54 for b = 1815 s/mm² and 0.47–0.51 for b = 5018 s/mm²). Simulations, phantoms and in vivo data indicate that greater signal attenuation, to a degree, is desirable for the accurate quantification of diffusion‐weighted spectra for slowly diffusing metabolites. Copyright © 2010 John Wiley & Sons, Ltd.     

Proton and phosphorus magnetic resonance spectroscopy of the healthy human breast at 7 T

In vivo water‐ and fat‐suppressed ¹H magnetic resonance spectroscopy (MRS) and ³¹P magnetic resonance adiabatic multi‐echo spectroscopic imaging were performed at 7 T in duplicate in healthy fibroglandular breast tissue of a group of eight volunteers. The transverse relaxation times of ³¹P metabolites were determined, and the reproducibility of ¹H and ³¹P MRS was investigated. The transverse relaxation times for phosphoethanolamine (PE) and phosphocholine (PC) were fitted bi‐exponentially, with an added short T₂ component of 20 ms for adenosine monophosphate, resulting in values of 199 ± 8 and 239 ± 14 ms, respectively. The transverse relaxation time for glycerophosphocholine (GPC) was also fitted bi‐exponentially, with an added short T₂ component of 20 ms for glycerophosphatidylethanolamine, which resonates at a similar frequency, resulting in a value of 177 ± 6 ms. Transverse relaxation times for inorganic phosphate, γ‐ATP and glycerophosphatidylcholine mobile phospholipid were fitted mono‐exponentially, resulting in values of 180 ± 4, 19 ± 3 and 20 ± 4 ms, respectively. Coefficients of variation for the duplicate determinations of ¹H total choline (tChol) and the ³¹P metabolites were calculated for the group of volunteers. The reproducibility of inorganic phosphate, the sum of phosphomonoesters and the sum of phosphodiesters with ³¹P MRS imaging was superior to the reproducibility of ¹H MRS for tChol. ¹H and ³¹P data were combined to calculate estimates of the absolute concentrations of PC, GPC and PE in healthy fibroglandular tissue, resulting in upper limits of 0.1, 0.1 and 0.2 mmol/kg of tissue, respectively.     

Precision and accuracy of diffusion kurtosis estimation and the influence of b‐value selection

Diffusion kurtosis imaging (DKI) is an extension of diffusion tensor imaging that accounts for leading non‐Gaussian diffusion effects. In DKI studies, a wide range of different gradient strengths (b‐values) is used, which is known to affect the estimated diffusivity and kurtosis parameters. Hence there is a need to assess the accuracy and precision of the estimated parameters as a function of b‐value. This work examines the error in the estimation of mean of the kurtosis tensor (MKT) with respect to the ground truth, using simulations based on a biophysical model for both gray (GM) and white (WM) matter. Model parameters are derived from densely sampled experimental data acquired in ex vivo rat brain and in vivo human brain. Additionally, the variability of MKT is studied using the experimental data. Prevalent fitting protocols are implemented and investigated. The results show strong dependence on the maximum b‐value of both net relative error and standard deviation of error for all of the employed fitting protocols. The choice of b‐values with minimum MKT estimation error and standard deviation of error was found to depend on the protocol type and the tissue. Protocols that utilize two terms of the cumulant expansion (DKI) were found to achieve minimum error in GM at b‐values less than 1 ms/μm², whereas maximal b‐values of about 2.5 ms/μm² were found to be optimal in WM. Protocols including additional higher order terms of the cumulant expansion were found to provide higher accuracy for the more commonly used b‐value regime in GM, but were associated with higher error in WM. Averaged over multiple voxels, a net average error of around 15% for both WM and GM was observed for the optimal b‐value choice. These results suggest caution when using DKI generated metrics for microstructural modeling and when comparing results obtained using different fitting techniques and b‐values.     

Multiband diffusion‐weighted MRI of the eye and orbit free of geometric distortions using a RARE‐EPI hybrid

Diffusion‐weighted imaging (DWI) provides information on tissue microstructure. Single‐shot echo planar imaging (EPI) is the most common technique for DWI applications in the brain, but is prone to geometric distortions and signal voids. Rapid acquisition with relaxation enhancement [RARE, also known as fast spin echo (FSE)] imaging presents a valuable alternative to DWI with high anatomical accuracy. This work proposes a multi‐shot diffusion‐weighted RARE‐EPI hybrid pulse sequence, combining the anatomical integrity of RARE with the imaging speed and radiofrequency (RF) power deposition advantage of EPI. The anatomical integrity of RARE‐EPI was demonstrated and quantified by center of gravity analysis for both morphological images and diffusion‐weighted acquisitions in phantom and in vivo experiments at 3.0 T and 7.0 T. The results indicate that half of the RARE echoes in the echo train can be replaced by EPI echoes whilst maintaining anatomical accuracy. The reduced RF power deposition of RARE‐EPI enabled multiband RF pulses facilitating simultaneous multi‐slice imaging. This study shows that diffusion‐weighted RARE‐EPI has the capability to acquire high fidelity, distortion‐free images of the eye and the orbit. It is shown that RARE‐EPI maintains the immunity to B₀ inhomogeneities reported for RARE imaging. This benefit can be exploited for the assessment of ocular masses and pathological changes of the eye and the orbit.     

Investigation of breast cancer using two‐dimensional MRS

Proton (¹H) MRS enables non‐invasive biochemical assay with the potential to characterize malignant, benign and healthy breast tissues. In vitro studies using perchloric acid extracts and ex vivo magic angle spinning spectroscopy of intact biopsy tissues have been used to identify detectable metabolic alterations in breast cancer. The challenges of ¹H MRS in vivo include low sensitivity and significant overlap of resonances due to limited chemical shift dispersion and significant inhomogeneous broadening at most clinical magnetic field strengths. Improvement in spectral resolution can be achieved in vivo and in vitro by recording the MR spectra spread over more than one dimension, thus facilitating unambiguous assignment of metabolite and lipid resonances in breast cancer. This article reviews the recent progress with two‐dimensional MRS of breast cancer in vitro, ex vivo and in vivo. The discussion includes unambiguous detection of saturated and unsaturated fatty acids, as well as choline‐containing groups such as free choline, phosphocholine, glycerophosphocholine and ethanolamines using two‐dimensional MRS. In addition, characterization of invasive ductal carcinomas and healthy fatty/glandular breast tissues non‐invasively using the classification and regression tree (CART) analysis of two‐dimensional MRS data is reviewed. Copyright © 2008 John Wiley & Sons, Ltd.     

Characterization of hepatic fatty acids in mice with reduced liver fat by ultra‐short echo time 1H‐MRS at 14.1 T in vivo

Alterations in the hepatic lipid content (HLC) and fatty acid composition are associated with disruptions in whole body metabolism, both in humans and in rodent models, and can be non‐invasively assessed by ¹H‐MRS in vivo. We used ¹H‐MRS to characterize the hepatic fatty‐acyl chains of healthy mice and to follow changes caused by streptozotocin (STZ) injection. Using STEAM at 14.1 T with an ultra‐short T_E of 2.8 ms, confounding effects from T₂ relaxation and J‐coupling were avoided, allowing for accurate estimations of the contribution of unsaturated (UFA), saturated (SFA), mono‐unsaturated (MUFA) and poly‐unsaturated (PUFA) fatty‐acyl chains, number of double bonds, PU bonds and mean chain length. Compared with in vivo ¹H‐MRS, high resolution NMR performed in vitro in hepatic lipid extracts reported longer fatty‐acyl chains (18 versus 15 carbons) with a lower contribution from UFA (61 ± 1% versus 80 ± 5%) but a higher number of PU bonds per UFA (1.39 ± 0.03 versus 0.58 ± 0.08), driven by the presence of membrane species in the extracts. STZ injection caused a decrease of HLC (from 1.7 ± 0.3% to 0.7 ± 0.1%), an increase in the contribution of SFA (from 21 ± 2% to 45 ± 6%) and a reduction of the mean length (from 15 to 13 carbons) of cytosolic fatty‐acyl chains. In addition, SFAs were also likely to have increased in membrane lipids of STZ‐induced diabetic mice, along with a decrease of the mean chain length. These studies show the applicability of ¹H‐MRS in vivo to monitor changes in the composition of the hepatic fatty‐acyl chains in mice even when they exhibit reduced HLC, pointing to the value of this methodology to evaluate lipid‐lowering interventions in the scope of metabolic disorders. Copyright © 2015 John Wiley & Sons, Ltd.     

Diffusion behavior of cerebral metabolites of congenital portal systemic shunt mice assessed noninvasively by diffusion‐weighted 1H magnetic resonance spectroscopy

Diffusion‐weighted ¹H‐MRS (DW‐MRS) allows for noninvasive investigation of the cellular compartmentalization of cerebral metabolites. DW‐MRS applied to the congenital portal systemic shunt (PSS) mouse brain may provide specific insight into alterations of cellular restrictions associated with PSS in humans. At 14.1 T, adult male PSS and their age‐matched healthy (Ctrl) mice were studied using DW‐MRS covering b‐values ranging from 0 to 45 ms/μm² to determine the diffusion behavior of abundant metabolites. The remarkable sensitivity and spectral resolution, in combination with very high diffusion weighting, allowed for precise measurement of the diffusion properties of endogenous N‐acetyl‐aspartate, total creatine, myo‐inositol, total choline with extension to glutamine and glutamate in mouse brains, in vivo. Most metabolites had comparable diffusion properties in PSS and Ctrl mice, suggesting that intracellular distribution space for these metabolites was not affected in the model. The slightly different diffusivity of the slow decaying component of taurine (0.015 ± 0.003 μm²/ms in PSS vs 0.021 ± 0.002 μm²/ms in Ctrl, P < 0.05) might support a cellular redistribution of taurine in the PSS mouse brain.     

Lipid biomarkers of glioma cell growth arrest and cell death detected by 1H magic angle spinning MRS

Biomarkers of early response to treatment have the potential to improve cancer therapy by allowing treatment to be tailored to the individual. Alterations in lipids detected by in vivo MRS have been suggested as noninvasive biomarkers of cell stress and early indicators of cell death. An improved understanding of the relationship between MRS lipids and cell stress in vitro would aid in the translation of this technique into clinical use. Rat BT4C glioma cells were treated with 50 µ m cis‐dichlorodiammineplatinum II (cisplatin), a commonly used chemotherapeutic agent, and harvested at several time points up to 72 h. High‐resolution magic angle spinning ¹H MRS of cells was then performed on a 600‐MHz NMR spectrometer. The metabolites were quantified using a time domain fitting method, TARQUIN. Increases were detected in saturated and polyunsaturated fatty acid resonances early during the exposure to cisplatin. The fatty acid CH₂/CH₃ ratio was unaltered by treatment after allowing for contributions of macromolecules. Polyunsaturated fatty acids increased on treatment, with the group –CH = CH–CH₂–CH = CH– accounting for all the unsaturated fatty acid signals. Transmission electron microscopy, in addition to Nile red and 4',6‐diamino‐2‐phenylindole co‐staining, revealed that the lipid increase was associated with cytoplasmic neutral lipid droplets. Small numbers of apoptotic and necrotic cells were detected by trypan blue, annexin V–fluorescein isothiocyanate‐labelled flow cytometry and DNA laddering after up to 48 h of cisplatin exposure. Propidium iodide flow cytometry revealed that cells accumulated in the G1 stage of the cell growth cycle. In conclusion, an increase in the size of the lipid droplets is detected in morphologically viable cells during cisplatin exposure. ¹H MRS can detect lipid alterations during cell cycle arrest and progression of cell death, and has the potential to provide a noninvasive biomarker of treatment efficacy in vivo. Copyright © 2012 John Wiley & Sons, Ltd.     

Correlation of Choline/Creatine and Apparent Diffusion Coefficient values with the prognostic parameters of Head and Neck Squamous Cell Carcinoma

The aim of this study was to measure choline/creatine (Ch/Cr) levels through ¹H‐MRS and apparent diffusion coefficient (ADC) values through diffusion‐weighted MRI, and to correlate these values with the prognostic parameters of head and neck squamous cell carcinoma (HNSCC). The institutional review board approved this study and informed written consent was obtained from all study participants. A prospective study of 43 patients (31 men and 12 women; mean age, 65 years) with HNSCC was conducted. Single‐voxel ¹H‐MRS was performed at the tumor or metastatic cervical lymph node with point‐resolved spectroscopy (PRESS) at TE = 135 ms. Diffusion‐weighted MR images with b values of 0, 500 and 1000 s/mm² and contrast MRI of the head and neck were performed. The Ch/Cr levels and ADC values of HNSCC were calculated. The gross tumor volume (GTV) was also calculated. The degree of tumor differentiation was determined through pathological examination. The HNSCC Ch/Cr level was negatively correlated with the ADC value (r = ?0.662, p = 0.001). There was a significant difference in the Ch/Cr and ADC values at different degrees of tumor differentiation (p = 0.003 and p = 0.001) and with different GTVs (p = 0.122 and p = 0.001). The following prognostic parameter categories were used: (i) poorly differentiated and undifferentiated versus well differentiated to moderately differentiated; and (ii) HNSCC with GTV < 30 cm³ versus GTV > 30 cm³. The cut‐off values for Cho/Cr and ADC for each category were 1.83, 0.95 and 1.94, 0.99, respectively, and the areas under the curve were 0.771, 0.967 and 0.726, 0.795, respectively, for each category. We conclude that the Ch/Cr levels determined using ¹H‐MRS and the ADC values are well correlated with several prognostic parameters of HNSCC. Copyright © 2016 John Wiley & Sons, Ltd.     

Detection of cancer in cervical tissue biopsies using mobile lipid resonances measured with diffusion‐weighted 1H magnetic resonance spectroscopy

The purpose of this study was to implement a diffusion‐weighted sequence for visualisation of mobile lipid resonances (MLR) using high resolution magic angle spinning (HR‐MAS) ¹H MRS and to evaluate its use in establishing differences between tissues from patients with cervical carcinoma that contain cancer from those that do not. A stimulated echo sequence with bipolar gradients was modified to allow T₁ and T₂ measurements and optimised by recording signal loss in HR‐MAS spectra as a function of gradient strength in model lipids and tissues. Diffusion coefficients, T₁ and apparent T₂ relaxation times were measured in model lipid systems. MLR profiles were characterised in relation to T₁ and apparent T₂ relaxation in human cervical cancer tissue samples. Diffusion‐weighted (DW) spectra of cervical biopsies were quantified and peak areas analysed using linear discriminant analysis (LDA). The optimised sequence reduced spectral overlap by suppressing signals originating from low molecular weight metabolites and non‐lipid contributions. Significantly improved MLR visualisation allowed visualisation of peaks at 0.9, 1.3, 1.6, 2.0, 2.3, 2.8, 4.3 and 5.3 ppm. MLR analysis of DW spectra showed at least six peaks arising from saturated and unsaturated lipids and those arising from triglycerides. Significant differences in samples containing histologically confirmed cancer were seen for peaks at 0.9 (p < 0.006), 1.3 (p < 0.04), 2.0 (p < 0.03), 2.8 (p < 0.003) and 4.3 ppm (p < 0.0002). LDA analysis of MLR peaks from DW spectra almost completely separated two clusters of cervical biopsies (cancer, ‘no‐cancer’), reflecting underlying differences in MLR composition. Generated Receiver Operating Characteristic (ROC) curves and calculated area under the curve (0.962) validated high sensitivity and specificity of the technique. Diffusion‐weighting of HR‐MAS spectroscopic sequences is a useful method for characterising MLR in cancer tissues and displays an accumulation of lipids arising during tumourigenesis and an increase in the unsaturated lipid and triglyceride peaks with respect to saturated MLR. Copyright © 2009 John Wiley & Sons, Ltd.     

In vivo and ex vivo evidence for ketamine-induced hyperglutamatergic activity in the cerebral cortex of the rat: Potential relevance to schizophrenia

Subanesthetic doses of ketamine, a noncompetitive N‐methyl‐D ‐aspartate (NMDA) receptor antagonist, impair prefrontal cortex (PFC) function in the rat and produce symptoms in humans similar to those observed in patients with schizophrenia. In the present study, in vivo ¹H‐MRS and ex vivo ¹H high‐resolution magic angle spinning (HR‐MAS) spectroscopy was used to examine the brain metabolism of rats treated with subanesthetic doses of ketamine (30 mg/kg) for 6 days. A single voxel localization sequence (PRESS, TR/TE = 4000/20 ms and NEX = 512) was used to acquire the spectra in a 30‐µl voxel positioned in the cerebral cortex (including mainly PFC) of the rats (ketamine group: n = 12; saline group: n = 12) anesthetized with isoflurane. After the in vivo ¹H‐MRS acquisition, the animals were sacrificed and the cerebral cortex tissues were extracted (ketamine group: n = 7; saline group: n = 7) for ex vivo ¹H HR‐MAS spectroscopy (CPMG sequence, 2.0‐s presaturation delay, 2.0‐s acquisition time, 128 transients and 4‐ms inter‐pulse delay) using a 500‐MHz NMR spectrometer. All proton metabolites were quantified using the LCModel. For the in vivo spectra, there was a significant increase in glutamate concentration in the cerebral cortex of the ketamine group compared with the controls (p < 0.05). For the ex vivo HR‐MAS spectra, there was a significant increase in the glutamate/total creatine ratio, and a decrease in the glutamine/total creatine and glutamine/glutamate ratios in the cerebral cortex tissue of the ketamine group compared with the controls. The results of the present study demonstrated that administration of subanesthetic doses of ketamine in the rat may exert at least part of their effect in the cerebral cortex by activation of glutamatergic neurotransmission. Copyright © 2011 John Wiley & Sons, Ltd.     

Cardiac lipid levels show diurnal changes and long‐term variations in healthy human subjects

¹H‐MRS is regularly applied to determine lipid content in ectopic tissue – mostly skeletal muscle and liver – to investigate physiological and/or pathologic conditions, e.g. insulin resistance. Technical developments also allow non‐invasive in vivo assessment of cardiac lipids; however, basic data about methodological reliability (repeatability) and physiological variations are scarce. The aim of the presented work was to determine potential diurnal changes of cardiac lipid stores in humans, and to put the results in relation to methodological repeatability and normal physiological day‐to‐day variations. Optimized cardiac‐ and respiratory‐gated ¹H‐MRS was used for non‐invasive quantification of intracardiomyocellular lipids (ICCL), creatine, trimethyl‐ammonium compounds (TMA), and taurine in nine healthy young men at three time points per day on two days separated by one week. This design allowed determination of (a) diurnal changes, (b) physiological variation over one week and (c) methodological repeatability of the ICCL levels. Comparison of fasted morning to post‐absorptive evening measurements revealed a significant 37 ± 19% decrease of ICCL during the day (p = 0.0001). There was a significant linear correlation between ICCL levels in the morning and their decrease during the day (p = 0.015). Methodological repeatability for the ICCL/creatine ratio was excellent, with a coefficient of variance of ~5%, whereas physiological variation was found to be considerably higher (22%) in spite of a standardized physiological preparation protocol. In contrast, TMA levels remained stable over this time period. The proposed ¹H‐MRS technique provides a robust way to investigate relevant physiological changes in cardiac metabolites, in particular ICCL. The present results suggest that ICCL reveal a diurnal course, with higher levels in the morning as compared to evening. In addition, a considerable long‐term variation of ICCL levels, in both the morning and evening, was documented. Given the high methodological repeatability, these effects should be taken into account in studies investigating the metabolic role of ICCL. Copyright © 2014 John Wiley & Sons, Ltd.