Probing regional glycogen metabolism in humans non-invasively has been challenging due to a lack of sensitive approaches. Here we studied human muscle glycogen dynamics post-exercise with a spatial resolution of millimeters and temporal resolution of minutes, using relayed nuclear Overhauser effect (glycoNOE) MRI. Data at 5T showed a homogeneous distribution of glycogen in resting muscle, with an average concentration of 99 ± 13 mM. After plantar flexion exercise following fasting with recovery under fasting conditions, the calf muscle showed spatially heterogeneous glycogen depletion and repletion kinetics that correlated with the severity of this depletion. Three types of regional glycogen kinetics were observed: (i) single exponential repletion (type a); (ii) biphasic recovery of rapid repletion followed by additional depletion (type b); (iii) biphasic recovery where continued depletion is followed by an exponential recovery (type c). The study of the complex patterns of glycogen kinetics suggests that glycogen breakdown may be quantitatively important during the initial recovery.

Conventional amide proton transfer (APT)-weighted imaging requires a chemical exchange saturation transfer (CEST) sequence with multiple saturation frequency offsets and a B0 correction sequence, plus a long acquisition time that can be reduced by applying the conventional method using CEST images with seven radiation pulses (i.e., the seven-points method). For a further reduction of acquisition times, we propose fast two-dimensional (2D) APT-weighted imaging based on a self B0 correction using the turbo spin echo (TSE)-Dixon method. We conducted a phantom study to investigate the accuracy of TSE-Dixon APT-weighted imaging.

We prepared two types of phantoms with six samples for a concentrationdependent evaluation and a pH-dependent evaluation. APT-weighted images were acquired by the conventional, seven-points, and TSE-Dixon methods. Linear regression analyses assessed the dependence between each method's APT signal intensities (SIs) and the concentration or pH. We performed a one-way analysis of variance with Tukey's honestly significant difference post hoc test to compare the APT SIs among the three methods. The agreement of the APT SIs between the conventional and seven-points or TSE-Dixon methods was assessed by a Bland- Altman plot analysis.

The APT SIs of all three acquisition methods showed positive concentration dependence and pH dependence. No significant differences were observed in the APT SIs between the conventional and TSE-Dixon methods at each concentration. The Bland-Altman plot analyses showed that the APT SIs measured with the seven-points method resulted in 0.42% bias and narrow 95% limits of agreement (LOA) (0.93%-0.09%) compared to the conventional method. The APT SIs measured using the TSE-Dixon method showed 0.14% bias and similar 95% LOA (-0.33% to 0.61%) compared with the seven-points method. The APT SIs of all three methods showed positive pH dependence. At each pH, no significant differences in the APT SIs were observed among the methods. Bland-Altman plot analyses showed that the APT SIs measured with the seven-points method resulted in low bias (0.03%) and narrow 95% LOA (-0.30% to 0.36%) compared to the conventional method. The APT SIs measured by the TSE-Dixon method showed slightly larger bias (0.29%) and similar 95% LOA (from -0.15% to 0.72%) compared to those measured by the seven-points method.

These results demonstrated that our proposed method has the same concentration dependence and pH dependence as the conventional method and the seven-points method. We thus expect that APT-weighted imaging with less influence of motion can be obtained in clinical examinations.

With the continuous innovation of magnetic resonance imaging (MRI) hardware and software technology, amide proton transfer-weighted (APTw) imaging has been applied in liver cancer. However, to our knowledge, no study has evaluated the feasibility of a three-dimensional amide proton transfer-weighted (3D-APTw) imaging sequence for hepatocellular carcinoma (HCC). This study thus aimed to conduct an image quality assessment of 3D-APTw for HCC and to explore its feasibility.

3D-APTw MRI examinations were completed in 134 patients with clinically suspected HCC. According to the uniformity of APTw signal in the liver and within the lesion and the proportion of artifact and missing signal regions, APTw images were subjectively scored using a 5-point scale. The scanning success rate of liver APTw imaging was calculated as the ratio of the number of cases with a quality assurance measurement of more than 3 to the total number of HCC cases. The intra- and interobserver quality assurance measurements for APTw images were compared via the Kappa consistency test. Within the HCC cases with a minimum image quality threshold of 3 points, the APT values of HCC and the liver parenchyma, signal-to-noise ratio of APT-weighted images (SNRAPTw), and contrast-to-noise ratio of HCC (CNRHCC) were measured by two observers. The intra- and interobserver agreement was assessed using the intraclass correlation coefficient (ICC). The differences in APT values between HCC and liver parenchyma was determined using the Mann-Whitney test.

Sixty-six HCC cases with a quality assurance measurement of APTw imaging were included in the final analysis, and the calculated success rate was 70.21% (66/94). The subjective APT image quality scores of the two observers were consistent (3.66±1.18, 3.50±1.19, and 3.68±1.18), and no intergroup or intragroup statistical differences were found (P=0.594, and P=0.091), but the consistency of inter- and intraobserver was not as satisfactory (κ=0.594 and κ=0.580). The APT values in HCC lesion were significantly higher than those in liver parenchyma (2.73%±0.91% vs. 1.62%±0.55%; P<0.001). The APT values in HCC showed favorable intra- and interobserver consistency between the two observers (ICC =0.808 and ICC =0.853); the APT values in liver parenchyma, SNRAPTw, and CNRHCC values had moderate intraobserver consistency (ICC =0.578, ICC =0.568, and ICC =0.508) and interobserver consistency (ICC =0.599, ICC =0.199, and ICC =0.650). The coefficients of variation of the APTw values in the HCC lesion and in liver parenchyma were 33.4% and 34.4%, respectively. The SNRAPTw and CNRHCC were 30.75±18.74 and 3.56±3.19, with a coefficient of variation of 60.9% and 74.9%, respectively.

Liver 3D-APTw imaging was preliminarily demonstrated to be clinically feasible for evaluating HCC.

This review aimed to perform a scoping review of promising MRI methods in assessing tumor hypoxia in hepatocellular carcinoma (HCC). The hypoxic microenvironment and upregulated hypoxic metabolism in HCC are determining factors of poor prognosis, increased metastatic potential, and resistance to chemotherapy and radiotherapy. Assessing hypoxia in HCC is essential for personalized therapy and predicting prognoses. Oxygen electrodes, protein markers, optical imaging, and positron emission tomography can evaluate tumor hypoxia. These methods lack clinical applicability because of invasiveness, tissue depth, and radiation exposure. MRI methods, including blood oxygenation level-dependent, dynamic contrast-enhanced MRI, diffusion-weighted imaging, MRI spectroscopy, chemical exchange saturation transfer MRI, and multinuclear MRI, are promising noninvasive methods that evaluate the hypoxic microenvironment by observing biochemical processes in vivo, which may inform on therapeutic options. This review summarizes the recent challenges and advances in MRI techniques for assessing hypoxia in HCC and highlights the potential of MRI methods for examining the hypoxic microenvironment via specific metabolic substrates and pathways. Although the utilization of MRI methods for evaluating hypoxia in patients with HCC is increasing, rigorous validation is needed in order to translate these MRI methods into clinical use. Due to the limited sensitivity and specificity of current quantitative MRI methods, their acquisition and analysis protocols require further improvement. EVIDENCE LEVEL: 3. TECHNICAL EFFICACY: Stage 4.

To develop a novel 3D abdominal CEST MRI technique at 3 T using MR multitasking, which enables entire-liver coverage with free-breathing acquisition.

k-Space data were continuously acquired with repetitive steady-state CEST (ss-CEST) modules. The stack-of-stars acquisition pattern was used for k-space sampling. MR multitasking was used to reconstruct motion-resolved 3D CEST images of 53 frequency offsets with entire-liver coverage and 2.0 × 2.0 × 6.0 mm3 spatial resolution. The total scan time was 9 min. The sensitivity of amide proton transfer (APT)-CEST (magnetization transfer asymmetry [MTRasym ] at 3.5 ppm) and glycogen CEST (glycoCEST) (mean MTRasym around 1.0 ppm) signals generated with the proposed method were tested with fasting experiments.

Both APT-CEST and glycoCEST signals showed high sensitivity between post-fasting and post-meal acquisitions. APT-CEST and glycoCEST MTRasym signals from post-mean scans were significantly increased (APT-CEST: -0.019 ± 0.017 in post-fasting scans, 0.014 ± 0.021 in post-meal scans, p < 0.01; glycoCEST: 0.003 ± 0.009 in post-fasting scans, 0.027 ± 0.021 in post-meal scans, p < 0.01).

The proposed 3D abdominal steady-state CEST method using MR multitasking can generate CEST images of the entire liver during free breathing.

All substances exert magnetic properties in some extent when placed in an external magnetic field. Magnetic susceptibility represents a measure of the magnitude of magnetization of a certain substance when the external magnetic field is applied. Depending on the tendency to be repelled or attracted by the magnetic field and in the latter case on the magnitude of this effect, materials can be classified as diamagnetic or paramagnetic, superparamagnetic and ferromagnetic, respectively. Knowledge of type and extent of susceptibility of common endogenous and exogenous substances and how their magnetic properties affect the conventional sequences used in magnetic resonance imaging (MRI) can help recognize them and exalt or minimize their presence in the acquired images, so as to improve diagnosis in a wide variety of benign and malignant diseases. Furthermore, in the context of diamagnetic susceptibility, chemical shift imaging enables to assess the intra-voxel ratio between water and fat content, analyzing the tissue composition of various organs and allowing a precise fat quantification. The following article reviews the fundamental physical principles of magnetic susceptibility and examines the magnetic properties of the principal endogenous and exogenous substances of interest in MRI, providing potential through representative cases for improved diagnosis in daily clinical routine.

The feasibility and image quality of three-dimensional (3D) amide proton transfer (APT)-weighted (APTw) in parotid tumor lesions have not been well established in previous studies. This study aimed to evaluate the utility of APT imaging in parotid lesions and glands.

Patients with parotid lesions received 3D turbo spin echo (TSE) APTw on a 3.0T scanner. Two radiologists, who were blinded to the clinical data, independently evaluated the APTw image quality using 4-point Likert scales (1= poor, 4= excellent) in terms of integrity and hyperintensity artifacts. An image quality selection protocol was built based on the two scores. Evaluable images (integrity score >1) and trustable images (integrity score >3 and hyperintensity artifacts score >2) were then enrolled for APTw value comparison between parotid lesions and glands.

Forty consecutive patients were included in this study. Four patients were excluded due to severe motion (n=3) or dental (n=1) artifacts, and 36 patients received the APT sequence. Among these, more parotid tumor lesions (34/36, 94.4%) than normal parotid glands (23/31, 74.2%) revealed excellent integrity scores (score =4) (P=0.034). Most parotid tumor lesions (24/34, 70.6%) and glands (16/28, 57.1%) revealed no or little hyperintensity artifacts for diagnosis (scores 3 and 4). APT values of parotid lesions and glands in the evaluable groups were 2.11%±1.15% and 1.60%±1.56%, respectively, and the difference was not significant (P=0.197). APT values of parotid lesions and glands in the trustable groups were 1.99%±1.18% and 1.03%±1.09%, respectively, and the difference was statistically significant (P=0.018).

3D APTw could be used to differentiate parotid tumors and normal parotid glands; however, the technology still needs to be improved to remove artifacts. In our study, most APTw images of tumor lesions in parotid glands had acceptable image quality, and these APTw images are feasible for diagnostic use.

An imaging method that allows quantitative fibrosis estimates is needed to facilitate the diagnosis of chronic liver disease. Amide proton transfer (APT) and tissue sodium concentration (TSC) estimates could meet this need.

APT and TSC estimates correlate with fibrosis in a mouse model of chronic liver disease.

Prospective.

Male C57Bl/6 mice given CCl₄ or vehicle (N = 8 each) twice weekly for 16 weeks.

Liver T₁ (Look-Locker gradient recalled echo [GRE] sequence), T₂ (multiecho spin echo sequence), T_1rho (fast spin echo sequence with 500 Hz spin locking pulse), and APT (GRE sequence with off-resonance pulses) data were acquired at 7 T at 12 and 16 weeks. Liver sodium data (multiple echo GRE sequence) were acquired at 12 weeks at 9.4 T.

Liver proton T₁ , T₂ , T_1rho , APT, sodium T₂ *, and TSC were calculated. Histological measures included Sirius Red, hematoxylin and eosin, liver hydroxyproline content, and serum alanine transaminase (ALT).

Welch's two-sided t-test was used to test for differences between control and CCl₄ -treated groups for serum ALT, hydroxyproline, Sirius Red staining, T₁ , T₂ , T_1rho , APT, TSC, and sodium T₂ *. Pearson's correlations between liver T₁ , APT, TSC, or sodium T₂ * with Sirius Red staining and hydroxyproline levels were calculated.

APT was significantly different (P < 0.05) between groups in the left liver lobe at 16 weeks (CCl₄ : 8.0% ± 1.2%, controls: 6.2% ± 1.0%), as were average liver TSC at 12 weeks (CCl₄ : 38 mM ± 5 mM, controls: 27 mM ± 2 mM), and average sodium liver T₂ * at 12 weeks (CCl₄ : 10 msec ± 1.0 msec, controls: 12 msec ± 1.9 msec). APT, TSC, and sodium T₂ * correlated significantly (P < 0.05) with Sirius Red staining and hydroxyproline levels.

Liver TSC and APT significantly correlated with histopathologic markers of fibrosis in this mouse model.

1 TECHNICAL EFFICACY: Stage 3.

There are a variety of less common diffuse liver diseases that can be asymptomatic or cause severe liver dysfunction. For the majority of them, the association of clinical, laboratory, and imaging findings are needed to narrow the differential diagnosis. In this article, we will review and describe the rarer diffuse liver diseases including drug-related liver disease, inflammatory and infectious diseases, and deposition disorders such as amyloidosis, glycogen storage disease, Wilson's disease, and alpha-1 antitrypsin deficiency. Abdominal radiologists should be familiar with the imaging features of different types of diffuse liver diseases to help the multidisciplinary team involved in the treatment of these patients. The data related to some of these conditions are scarce and sometimes experimental, but we want to demonstrate to the reader the value of imaging techniques in their analysis and introduce the potential of new imaging methods.

To investigate the feasibility of using amide proton transfer (APT) magnetic resonance imaging (MRI) in the liver and to evaluate its ability to characterize focal liver lesions (FLLs).

A total of 203 patients with suspected FLLs who underwent APT imaging at 3T were included. APT imaging was obtained using a single-slice turbo spin-echo sequence to include FLLs through five breath-holds, and its acquisition time was approximately 1 min. APT signals in the background liver and FLL were measured with magnetization transfer ratio asymmetry (MTR_asym) at 3.5 ppm. The technical success rate of APT imaging and the reasons for failure to obtain meaningful MTR_asym values were assessed. The Mann Whitney U test was used to compare MTR_asym values between different FLLs.

The technical success rate of APT imaging in the liver was 62.1% (126/203). The reasons for failure were a too large B₀ inhomogeneity (n = 43), significant respiratory motion (n = 12), and these two factors together (n = 22), respectively. Among 59 FLLs with analyzable APT images, MTR_asym values were compared between 27 patients with liver metastases and 23 patients with hepatocellular carcinomas (HCCs). The MTR_asym values of metastases were significantly higher than those of HCC (0.13 ± 2.15% vs. - 1.41 ± 3.68%, p = 0.027).

APT imaging could be an imaging biomarker for the differentiation of FLLs. However, further technical improvement is required before APT imaging can be clinically applied to liver MRI.

• Liver APT imaging was technically feasible, but with a relatively low success rate (62.1%). • Liver metastases showed higher APT values than hepatocellular carcinomas.

Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver, preoperative grading of HCC is of great clinical significance. Amide proton transfer-weighted (APTw) imaging, as a novel contrast mechanism in the field of molecular imaging, provided new diagnostic ideas for the grading of HCC.

Between May 2017 and April 2018, 32 consecutive patients with pathologically confirmed HCC were enrolled, including 19 high-grade HCCs and 13 low-grade HCCs. DWI and APTw scanning was performed on a 3T MRI scanner. Two observers drew regions of interest independently by referring to the axial T₂-weighted imaging, and APTw and apparent diffusion coefficient (ADC) values were obtained. Inter- and intra-observer agreements were assessed with the intraclass correlation coefficients (ICCs). The independent sample t test was used to compare the APTw and ADC values between the high- and low-grade HCC tumor parenchyma. The receiver operating characteristic curve was used to analyze the diagnostic efficacy of high- from low-grade HCC tumors. Spearman correlation analysis was used to assess the relationship between APTw and ADC values and HCC histological grades.

There were significant differences between the APTw or ADC values for the high- and low-grade HCCs (P=0.034 and 0.010). Both APTw and DWI had good diagnostic performance in differentiating the high- from the low-grade HCCs, with areas under the curves of 0.814 and 0.745, respectively. Moderate correlations existed between APTw values and histological grades (r=0.534; P=0.002), as well as ADC values and histological grades (r=-0.417; P=0.018).

The APTw imaging is a useful imaging biomarker that complements DWI for the more accurate and comprehensive HCC characterization.

To prospectively evaluate the feasibility and capability of amide proton transfer-weighted (APTw) imaging for the characterization of head and neck tumors.

Twenty-nine consecutive patients with suspected head and neck tumors were enrolled in this study and underwent APTw magnetic resonance imaging (MRI) on a 3.0-T MRI scanner. The patients were divided into malignant (n = 16) and benign (n = 13) groups, based on pathological results. A map of magnetization transfer ratio asymmetry at 3.5 ppm [MTR_asym (3.5 ppm)] was generated for each patient. Interobserver agreement was evaluated and comparisons of MTR_asym (3.5 ppm) were made between the malignant and benign groups. Receiver operating characteristic analysis was used to determine the appropriate threshold value of MTR_asym (3.5 ppm) for the differentiation of malignant from benign tumors.

The intraclass correlation coefficients of the malignant and benign groups were 0.96 and 0.90, respectively, which indicated a good interobserver agreement. MTR_asym (3.5 ppm) was significantly higher for the malignant group (3.66 ± 1.15 %) than for the benign group (1.94 ± 0.93 %, P < 0.001). APTw MRI revealed an area under the curve of 0.904 in discriminating these two groups, with a sensitivity of 81.3 %, a specificity of 92.3 %, and an accuracy of 86.2 %, at the threshold of 2.62 % of MTR_asym (3.5 ppm).

APTw MRI is feasible for use in the head and neck tumors and is a valuable imaging biomarker for distinguishing malignant from benign lesions.

Sugar-based biopolymers have been recognized as attractive materials to develop macromolecule- and nanoparticle-based cancer imaging and therapy. However, traditional biopolymer-based imaging approaches rely on the use of synthetic or isotopic labeling, and because of it, clinical translation often is hindered. Recently, a novel magnetic resonance imaging (MRI) technology, chemical exchange saturation transfer (CEST), has emerged, which allows the exploitation of sugar-based biopolymers as MRI agents by their hydroxyl protons-rich nature. In the study, we reviewed recent studies on the topic of CEST MRI detection of sugar-based biopolymers. The CEST MRI property of each biopolymer was briefly introduced, followed by the pre-clinical and clinical applications. The findings of these preliminary studies imply the enormous potential of CEST detectable sugar-based biopolymers in developing highly sensitive and translatable molecular imaging agents and constructing image-guided biopolymer-based drug delivery systems. This article is categorized under: Diagnostic Tools > in vivo Nanodiagnostics and Imaging Therapeutic Approaches and Drug Discovery > Emerging Technologies.

Amide proton transfer-weighted (APTw) imaging is a molecular MRI technique that generates image contrast based predominantly on the amide protons in mobile cellular proteins and peptides that are endogenous in tissue. This technique, the most studied type of chemical exchange saturation transfer imaging, has been used successfully for imaging of protein content and pH, the latter being possible due to the strong dependence of the amide proton exchange rate on pH. In this article we briefly review the basic principles and recent technical advances of APTw imaging, which is showing promise clinically, especially for characterizing brain tumors and distinguishing recurrent tumor from treatment effects. Early applications of this approach to stroke, Alzheimer's disease, Parkinson's disease, multiple sclerosis, and traumatic brain injury are also illustrated. Finally, we outline the technical challenges for clinical APT-based imaging and discuss several controversies regarding the origin of APTw imaging signals in vivo. Level of Evidence: 3 Technical Efficacy Stage: 3 J. Magn. Reson. Imaging 2019;50:347-364.

Performing chemical exchange saturation transfer (CEST) magnetic resonance imaging (MRI) in lung tissue is difficult because of motion artifacts. We, therefore, developed a CEST MRI acquisition and analysis method that performs retrospective respiration gating. Our method used an acquisition scheme with a short 200-millisecond saturation pulse that can accommodate the timing of the breathing cycle, and with saturation applied at frequencies in 0.03-ppm intervals. The Fourier transform of each image was used to calculate the difference in phase angle between adjacent pixels in the longitudinal direction of the respiratory motion. Additional digital filtering techniques were used to evaluate the breathing cycle, which was used to construct CEST spectra from images during quiescent periods. Results from CEST MRI with and without respiration gating analysis were used to evaluate the asymmetry of the magnetization transfer ratio (MTR_asym), a measure of CEST, for an egg white phantom that underwent cyclic motion, in the liver of healthy patients, as well as liver and tumor tissues of patients diagnosed with lung cancer. Retrospective respiration gating analysis produced more precise measurements in all cases with significant motion compared with nongated analysis methods. Finally, a preliminary clinical study with the same respiration-gated CEST MRI method showed a large increase in MTR_asym after radiation therapy, a small increase or decrease in MTR_asym after chemotherapy, and mixed results with combined chemoradiation therapy. Therefore, our retrospective respiration-gated method can improve CEST MRI evaluations of tumors and organs that are affected by respiratory motion.

To explore black blood T1rho (T1ρ) liver imaging and investigate the earliest stage when biliary duct ligation (BDL) induced liver fibrosis can be diagnosed.

MR was performed at 3 Tesla. A T1ρ prepared 2D fast spin echo (FSE) sequence with acquisition of four spin lock times (TSLs: 1, 10, 30, and 50 msec) and spin-lock frequency of 500 Hz was applied. Inherent black blood effect of FSE and double inversion recovery (DIR) achieved blood signal suppression, and 3 axial sections per liver were obtained. Male Sprague-Dawley rats were scanned at baseline (n=32), and on day-3 (n=13), day-5 (n=11), day-7 (n=10), day-10 (n=4) respectively after BDL. Hematoxylin-eosin (HE) and picrosirius red staining liver histology was obtained at these time points.

The physiological liver parenchyma T1ρ was 38.38±1.53 msec (range, 36.05-41.53 msec). Liver T1ρ value elevated progressively after BDL. On day-10 after BDL all experimental animals can be separated from normal liver based on T1ρ measurement with lowest value being 42.82 msec. Day-7 and day-10 liver resembled METAVIR stage-F1/F2 fibrosis, and fibrous area counted for 0.22%±0.13% and 0.38%±0.44% of liver parenchyma area, respectively.

This study provides the first proof-of-principle that T1ρ might diagnose early stage liver fibrosis.

Citicoline (CDPC) is a natural supplement with well-documented neuroprotective effects in the treatment of neurodegenerative diseases. In the present study, we sought to exploit citicoline as a theranostic agent with its inherent chemical exchange saturation transfer (CEST) MRI signal, which can be directly used as an MRI guidance in the citicoline drug delivery. Our in vitro CEST MRI results showed citicoline has two inherent CEST signals at +1 and +2 ppm, attributed to exchangeable hydroxyl and amine protons, respectively. To facilitate the targeted drug delivery of citicoline to ischemic regions, we prepared liposomes encapsulating citicoline (CDPC-lipo) and characterized the particle properties and CEST MRI properties. The in vivo CEST MRI detection of liposomal citicoline was then examined in a rat brain model of unilateral transient ischemia induced by a two-hour middle cerebral artery occlusion. The results showed that the delivery of CPDC-lipo to the brain ischemic areas could be monitored and quantified by CEST MRI. When administered intra-arterially, CDPC-lipo clearly demonstrated a detectable CEST MRI contrast at 2 ppm. CEST MRI revealed that liposomes preferentially accumulated in the areas of ischemia with a disrupted blood-brain-barrier. We furthermore used CEST MRI to detect the improvement in drug delivery using CDPC-lipo targeted against vascular cell adhesion molecule (VCAM)-1 in the same animal model. The MRI findings were validated using fluorescence microscopy. Hence, liposomal citicoline represents a prototype theranostic system, where the therapeutic agent can be detected directly by CEST MRI in a label-free fashion.