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Gastrointestinal stromal tumors (GISTs) and malignant lymphomas (MLs) in the abdomen are often observed as tumors of unknown origin on F-18 FDG PET/CT. The purpose of this study was to evaluate the intratumoral metabolic heterogeneity of F-18 FDG uptake on PET to determine if it might be helpful to discriminate between these tumors.The F-18 FDG PET/CT findings of 21 large abdominal tumors were retrospectively evaluated (9 GISTs and 12 MLs). Intratumoral heterogeneity was evaluated by visual scoring (visual score: 0, homogeneous; 1, slightly heterogeneous; 2, moderately heterogeneous; 3, highly heterogeneous) and by the cumulative standardized uptake value (SUV) histograms on transaxial PET images at the maximal cross-sectional tumor diameter. Percent tumor areas above a threshold from 0 to 100% of the maximum SUV were plotted and the area under curve of the cumulative SUV histograms (AUC-CSH) was used as a heterogeneity index, where lower values corresponded with increased heterogeneity. Correlation between the visual score and the AUC-CSH was investigated by the Spearman's rank test.GISTs exhibited heterogeneous uptake of F-18 FDG, whereas MLs showed rather homogeneous uptake on visual analysis (visual score: 2.67 ± 0.50 and 0.58 ± 0.79, respectively; p 0.001). The AUC-CSH was significantly lower for the GISTs than for the MLs (0.41 ± 0.14 and 0.64 ± 0.08, respectively; p 0.001). Significant correlations were observed between the visual score and the AUC-CSH (ρ = -0.866, p 0.001).GISTs exhibited significantly heterogeneous intratumoral tracer uptake as compared with the MLs. Evaluation of the intratumoral heterogeneity of F-18 FDG uptake may help in the discrimination between these tumors.

Discriminating tumor infiltrative and vasogenic brain edema in malignant gliomas is important although challenging in clinical settings. This study challenged this issue by performing voxel-wise analysis of (18)F-fluorodeoxy glucose (FDG) and (11)C-methionine positron emission tomography (PET) in peritumoral brain edemas. The authors studied ten malignant glioma and nine meningioma patients with peritumoral brain edema. A voxel-wise analysis of FDG and (11)C-methionine PET was performed in order to quantify the correlation between uptake of these tracers in normal brain tissue and peritumoral brain edema. Decoupling score of the uptake of two tracers was calculated as the z-score from the estimated correlation between uptake of the two tracers in normal brain tissue. The decoupling score was also converted into images for visual inspection. Average decoupling score in the peritumoral brain edema was calculated and compared between those obtained from malignant gliomas and meningiomas. FDG and (11)C-methionine uptake showed a reproducible linear correlation in normal brain tissue. This correlation was preserved in peritumoral edema of meningioma, but not in that of malignant gliomas. In malignant gliomas, higher (11)C-methionine uptake compared to that estimated by the FDG uptake in normal brain tissue was observed, thus suggesting that decoupling was caused by tumor infiltration. Visual inspection of the decoupling score enabled discrimination of tumor infiltrative and vasogenic edema. The average decoupling scores of the peritumoral brain edema in malignant gliomas were significantly higher than those in meningiomas (2.9 vs. 0.7, P = 0.0003). As a conclusion, FDG/(11)C-methionine uptake decoupling score can be used for the discrimination of tumor infiltrative and vasogenic brain edema. The proposed method also suggests the possibility of accurately detecting tumor infiltration into brain tissues in gliomas, providing significant information for treatment planning and follow-up.

Background Boron neutron capture therapy (BNCT) is a molecular radiation treatment based on the 10B (n, α) 7Li nuclear reaction in cancer cells, in which delivery of 10B by 4-borono-phenylalanine conjugated with fructose (BPA-fr) to the cancer cells is of critical importance. The PET tracer 4-borono-2-18 F-fluoro-phenylalanine (FBPA) has been used to predict the accumulation of BPA-fr before BNCT. However, because of the difference in chemical structure between BPA-fr and FBPA and the difference in the dose administered between BPA-fr (therapeutic dose) and FBPA (tracer dose), the predictive value of FBPA PET for BPA-fr accumulation in the tumor and normal tissues is not yet clearly proven. We conducted this study to validate FBPA PET as a useful test to predict the accumulation of BPA-fr in the tumor and normal tissues before BNCT. Methods RGC-6 rat glioma cells (1.9 × 107) were implanted subcutaneously in seven male F344 rats. On day 20 after the tumor implantation, dynamic PET scan was performed on four rats after injection of FBPA for 1 h. Whole-body PET/CT was performed 1 h after intravenous injection of the FBPA solution (30.5 ± 0.7 MBq, 1.69 ± 1.21 mg/kg). PET accumulation of FBPA in the tumor tissue and various normal tissues was estimated as a percentage of the injected dose per gram (%ID/g). One hour after the PET/CT scan, BPA-fructose (167.32 ± 18.65 mg/kg) was injected intravenously, and the rats were dissected 1 h after the BPA-fr injection. The absolute concentration of 10B in the autopsied tissues and blood was measured by inductively coupled plasma optical emission spectrometry (ICP-OES). Results The highest absolute concentration of 10B determined by ICP-OES was found in the kidney (4.34 ± 0.84 %ID/g), followed by the pancreas (2.73 ± 0.63 %ID/g), and the tumor (1.44 ± 0.44 %ID/g). A significant positive correlation was found between the accumulation levels of BPA-fr and FBPA (r = 0.91, p Conclusions FBPA PET can reliably predict accumulation of BPA-fr in the tumor as well as normal tissues.

Paradoxical reduction of cerebral blood flow (CBF) after administration of the vasodilator acetazolamide is the most severe stage of cerebrovascular reactivity failure and is often associated with an increased oxygen extraction fraction (OEF). In this study, we aimed to reveal the mechanism underlying this phenomenon by focusing on the ratio of CBF to cerebral blood volume (CBV) as a marker of regional cerebral perfusion pressure (CPP). In 37 patients with unilateral internal carotid or middle cerebral arterial (MCA) steno-occlusive disease and 8 normal controls, the baseline CBF (CBF(b)), CBV, OEF, cerebral oxygen metabolic rate (CMRO2), and CBF after acetazolamide loading in the anterior and posterior MCA territories were measured by (15)O positron emission tomography. Paradoxical CBF reduction was found in 28 of 74 regions (18 of 37 patients) in the ipsilateral hemisphere. High CBF(b) (47.6 mL/100 mL/min, n = 7) was associated with normal CBF(b)/CBV, increased CBV, decreased OEF, and normal CMRO2. Low CBF(b) (31.8 mL/100 mL/min, n = 9) was associated with decreased CBF(b)/CBV, increased CBV, increased OEF, and decreased CMRO2. These findings demonstrated that paradoxical CBF reduction is not always associated with reduction of CPP, but partly includes high-CBF(b) regions with normal CPP, which has not been described in previous studies.

Local cerebral perfusion pressure (CPP) is a primary factor controlling cerebral circulation and previous studies have indicated that the ratio of cerebral blood flow (CBF) to cerebral blood volume (CBV) can be used as an index of the local CPP. In this study, we investigated whether the CBF/CBV ratio differs among different brain structures under physiological conditions, by means of (15)O positron emission tomography. Nine healthy volunteers (5 men and 4 women; mean age, 47.0 ± 1.2 years) were studied by H2 (15)O bolus injection for CBF measurement and by C(15)O inhalation for CBV measurement. The CBF/CBV ratio maps were created by dividing the CBF images by the CBV images after anatomical normalization. Regions of interest were placed on the CBF/CBV maps and comparing the regions. The mean CBF/CBV ratio was highest in the cerebellum (19.3 ± 5.2/min), followed by the putamen (18.2 ± 3.9), pons (16.4 ± 4.6), thalamus (14.5 ± 3.3), cerebral cortices (13.2 ± 2.4), and centrum semiovale (11.5 ± 2.1). The cerebellum and putamen showed significantly higher CBF/CBV ratios than the cerebral cortices and centrum semiovale. We created maps of the CBF/CBV ratio in normal volunteers and demonstrated higher CBF/CBV ratios in the cerebellum and putamen than in the cerebral cortices and deep cerebral white matter. The CBF/CBV may reflect the local CPP and should be studied in hemodynamically compromised patients and in patients with risk factors for small-artery diseases of the brain.

PET with (15)O gas has been used for the quantitative measurement of cerebral blood flow (CBF), cerebral metabolic rate of oxygen (CMRO(2)), oxygen extraction fraction (OEF), and cerebral blood volume (CBV) in humans. However, several technical difficulties limit its use in experiments on small animals. Herein, we describe the application of the (15)O gas steady-state inhalation method for normal anesthetized rats.Eight normal male Sprague-Dawley rats (mean body weight ± SD, 268 ± 14 g) under anesthesia were investigated by (15)O-labeled gas PET. After tracheotomy, an airway tube was placed in the trachea, and the animals were connected to a ventilator (tidal volume, 3 cm(3); frequency, 60/min). The CBF and OEF were measured according to the original steady-state inhalation technique under artificial ventilation with (15)O-CO(2) and (15)O-O(2) gases delivered through the radioactive gas stabilizer. CBV was measured by (15)O-CO gas inhalation and corrected for the intravascular hemoglobin-bound (15)O-O(2). Arterial blood sampling was performed during each study to measure the radioactivity of the whole blood and plasma. MR image was performed with the same acrylic animal holder immediately after the PET. Regions of interest were placed on the whole brain of the PET images with reference to the semiautomatically coregistered PET/MR fused images.The data acquisition time for the whole PET experiment in each rat was 73.3 ± 5.8 (range, 68-85) min. In both the (15)O-CO(2) and the (15)O-O(2) studies, the radioactivity count of the brain reached a steady state by approximately 10 min after the start of continuous inhalation of the gas. The quantitative PET data of the whole brain were as follows: CBF, 32.3 ± 4.5 mL/100 mL/min; CMRO(2), 3.23 ± 0.42 mL/100 mL/min; OEF, 64.6% ± 9.1%; and CBV, 5.05 ± 0.45 mL/100 mL.Although further technical improvements may be needed, this study demonstrated the feasibility of quantitative PET measurement of CBF, OEF, and CMRO(2) using the original steady-state inhalation method of (15)O-CO(2) and (15)O-O(2) gases and measurement of CBV using the (15)O-CO gas inhalation method in the brain of normal anesthetized rats.

Introduction Coronary microvascular dysfunction (MVD) has been shown to be the major cause of persistent myocardial ischaemia and progressive left ventricular (LV) remodelling in the chronic myocardial infarction (MI) heart. ONO-1301SR is a slow-releasing synthetic prostacyclin agonist, being developed to induce angiogenesis by upregulating a variety of proangiogenic cytokines in the targeted region. In addition, pedicle omental flap covering over the LV surface reportedly yields proangiogenic effects on chronic MI heart. We herein hypothesised pedicle omental flap covering, combined with ONO-1301SR placement. over the LV surface may be effective on the MVD and related functional deterioration in a chronic MI heart. To test this hypothesis, clinically latest imaging studies were applied in a porcine chronic MI model. Methods A mini-pig chronic MI model was generated by placing an ameroid constrictor around the left anterior descending artery for 4 weeks. The mini-pigs were then randomly assigned into the following 4 groups; sham group, omental flap alone (OM group), ONO-1301SR alone (ONO group), ONO-1301SR plus omental flap (ONO+OM group, n = 6 each). Results At 4 weeks after the treatment, 13 N-ammonia PET study showed that the ONO+OM group, but not the other groups, displayed a significantly increase in myocardial blood flow (189 ± 89% increase, P vascular endothelial growth factor, hepatocyte growth factor or stem cell-derived factor-1 , were not significantly different among the groups, indicating that angiogenic process might have been completed at the 4 weeks after the treatment. Conclusions ONO-1301SR combined with omental flap therapy promoted myocardial angiogenesis with collateral artery growth, leading to recovery of the cardiac function in a porcine ICM model. This combined therapy may be useful in regenerating myocardial microvasculature.

Immunotherapy targeting the Wilms tumor 1 (WT1) gene product is a promising treatment modality for patients with malignant gliomas, and there have been reports of encouraging results. It has become clear, however, that Gd-enhanced MR imaging does not reflect prognosis, thereby necessitating a more robust imaging evaluation system for monitoring response to WT1 immunotherapy. To meet this demand, the authors performed a voxel-wise parametric response map (PRM) analysis of (11)C-methionine PET (MET-PET) in WT1 immunotherapy and compared the data with the overall survival after initiation of WT1 immunotherapy (OS(WT1)).Fourteen patients with recurrent malignant glioma were included in the study, and OS(WT1) was compared with: 1) volume and length change in the contrast area of the tumor on Gd-enhanced MR images; 2) change in maximum uptake of (11)C-methionine; and 3) a more detailed voxel-wise PRM analysis of MET-PET pre- and post-WT1 immunotherapy.The PRM analysis was able to identify the following 3 areas within the tumor core: 1) area with no change in (11)C-methionine uptake pre- and posttreatment; 2) area with increased (11)C-methionine uptake posttreatment (PRM(+MET)); and 3) area with decreased (11)C-methionine uptake posttreatment. While the results of Gd-enhanced MR imaging volumetric and conventional MET-PET analysis did not correlate with OS(WT1) (p = 0.270 for Gd-enhanced MR imaging length, p = 0.960 for Gd-enhanced MR imaging volume, and p = 0.110 for MET-PET), the percentage of PRM(+MET) area showed excellent correlation (p = 0.008) with OS(WT1).This study describes the limited value of Gd-enhanced MR imaging and highlights the potential of voxel-wise PRM analysis of MET-PET for monitoring treatment response in immunotherapy for malignant gliomas. Clinical trial registration no.: UMIN000002001.

Discriminating tumor infiltrative and vasogenic brain edema in malignant gliomas is important although challenging in clinical settings. This study challenged this issue by performing voxel-wise analysis of (18)F-fluorodeoxy glucose (FDG) and (11)C-methionine positron emission tomography (PET) in peritumoral brain edemas. The authors studied ten malignant glioma and nine meningioma patients with peritumoral brain edema. A voxel-wise analysis of FDG and (11)C-methionine PET was performed in order to quantify the correlation between uptake of these tracers in normal brain tissue and peritumoral brain edema. Decoupling score of the uptake of two tracers was calculated as the z-score from the estimated correlation between uptake of the two tracers in normal brain tissue. The decoupling score was also converted into images for visual inspection. Average decoupling score in the peritumoral brain edema was calculated and compared between those obtained from malignant gliomas and meningiomas. FDG and (11)C-methionine uptake showed a reproducible linear correlation in normal brain tissue. This correlation was preserved in peritumoral edema of meningioma, but not in that of malignant gliomas. In malignant gliomas, higher (11)C-methionine uptake compared to that estimated by the FDG uptake in normal brain tissue was observed, thus suggesting that decoupling was caused by tumor infiltration. Visual inspection of the decoupling score enabled discrimination of tumor infiltrative and vasogenic edema. The average decoupling scores of the peritumoral brain edema in malignant gliomas were significantly higher than those in meningiomas (2.9 vs. 0.7, P = 0.0003). As a conclusion, FDG/(11)C-methionine uptake decoupling score can be used for the discrimination of tumor infiltrative and vasogenic brain edema. The proposed method also suggests the possibility of accurately detecting tumor infiltration into brain tissues in gliomas, providing significant information for treatment planning and follow-up.

To demonstrate that positron emission tomography (PET)/magnetic resonance imaging (MRI) fusion was feasible in characterizing pancreatic tumors (PTs), comparing MRI and computed tomography (CT) as mapping images for fusion with PET as well as fused PET/MRI and PET/CT.We retrospectively reviewed 47 sets of (18)F-fluorodeoxyglucose ((18)F -FDG) PET/CT and MRI examinations to evaluate suspected or known pancreatic cancer. To assess the ability of mapping images for fusion with PET, CT (of PET/CT), T1- and T2-weighted (w) MR images (all non-contrast) were graded regarding the visibility of PT (5-point confidence scale). Fused PET/CT, PET/T1-w or T2-w MR images of the upper abdomen were evaluated to determine whether mapping images provided additional diagnostic information to PET alone (3-point scale). The overall quality of PET/CT or PET/MRI sets in diagnosis was also assessed (3-point scale). These PET/MRI-related scores were compared to PET/CT-related scores and the accuracy in characterizing PTs was compared.Forty-three PTs were visualized on CT or MRI, including 30 with abnormal FDG uptake and 13 without. The confidence score for the visibility of PT was significantly higher on T1-w MRI than CT. The scores for additional diagnostic information to PET and overall quality of each image set in diagnosis were significantly higher on the PET/T1-w MRI set than the PET/CT set. The diagnostic accuracy was higher on PET/T1-w or PET/T2-w MRI (93.0 and 90.7%, respectively) than PET/CT (88.4%), but statistical significance was not obtained.PET/MRI fusion, especially PET with T1-w MRI, was demonstrated to be superior to PET/CT in characterizing PTs, offering better mapping and fusion image quality.

We compared the diagnostic accuracy of fluorodeoxyglucose positron emission tomography/computed tomography (FDG PET/CT) and PET/magnetic resonance imaging (MRI) fusion images for gynecological malignancies.A total of 31 patients with gynecological malignancies were enrolled. FDG-PET images were fused to CT, T1- and T2-weighted images (T1WI, T2WI). PET-MRI fusion was performed semiautomatically. We performed three types of evaluation to demonstrate the usefulness of PET/MRI fusion images in comparison with that of inline PET/CT as follows: depiction of the uterus and the ovarian lesions on CT or MRI mapping images (first evaluation); additional information for lesion localization with PET and mapping images (second evaluation); and the image quality of fusion on interpretation (third evaluation).For the first evaluation, the score for T2WI (4.68 +/- 0.65) was significantly higher than that for CT (3.54 +/- 1.02) or T1WI (3.71 +/- 0.97) (P0.01). For the second evaluation, the scores for the localization of FDG accumulation showing that T2WI (2.74 +/- 0.57) provided significantly more additional information for the identification of anatomical sites of FDG accumulation than did CT (2.06 +/- 0.68) or T1WI (2.23 +/- 0.61) (P0.01). For the third evaluation, the three-point rating scale for the patient group as a whole demonstrated that PET/T2WI (2.72 +/- 0.54) localized the lesion significantly more convincingly than PET/CT (2.23 +/- 0.50) or PET/T1WI (2.29 +/- 0.53) (P0.01).PET/T2WI fusion images are superior for the detection and localization of gynecological malignancies.

The aim of this study was to evaluate the diagnostic accuracy of positron emission tomography (PET) with 2-[18F]fluoro-2-deoxy-D-glucose (18F-FDG) for staging/restaging, evaluating the treatment response, and screening of recurrence in patients with malignant lymphoma (ML) during long-term follow-up, and to compare that with computer tomography (CT)/magnetic resonance imaging (MRI).The study was conducted in 59 ML patients who underwent whole-body 18F-FDG-PET examinations three times or more from October 1998 to August 2006. The location of the lesions in the patients with positive findings on 18F-FDG-PET and/or the corresponding CT/MRI was classified into supradiaphragmatic (n = 10), infradiaphragmatic (n = 7), and extranodal sites (n = 20), and the findings were compared on a site basis according to the gold standard, which consisted of all clinical information available, including follow-up results.A total of 156 18F-FDG-PET examinations for which the corresponding CT/MRI images were also available were evaluated, and a total of 305 sites showed positive findings on 18F-FDG-PET and/or CT/MRI. Concordant positive findings were obtained in 76% for staging/restaging, 34% for evaluation of the treatment response, and 50% for screening of recurrence. The accuracy of 18F-FDG-PET versus CT/MRI was 92% versus 84% (P = 0.06) for staging/restaging, 84% versus 50% (P0.05) for the evaluation of the treatment response, and 83% versus 72% (P = 0.21) for the screening of recurrence. At pathologic sites with discrepant findings between 18F-FDG-PET and CT/MRI (n = 122), the frequency of accurate diagnosis by 18F-FDG-PET (76%) was higher than that by CT/MRI (24%), especially for the evaluation of the treatment response.18F-FDG-PET is expected to play a significant role in the management of ML patients even after effective treatment is initiated.

The new RECIST criteria for evaluation of tumor response to chemotherapy and/or radiotherapy were proposed in 1999. We compared RECIST with the WHO criteria and also compared both methods with the histological findings, to evaluate RECIST.The subjects were 32 primary lung cancer patients operated on after chemotherapy and/or radiotherapy. Two radiologists measured the diameter of the tumors and compared the RECIST and WHO criteria using the McNemar test. We also compared both criteria with the histological results.Using RECIST, partial response (PR) was assessed in 12 cases, stable disease (SD) in 18, and progressive disease (PD) in 2. With the WHO criteria, PR was seen in 15, no change (NC) in 15, and PD in 2. The two evaluations corresponded in 29 of 32 cases, and the p-value was 0.2500. Ef.1b-2 (good histological effect) was much higher in PR, and to evaluate the size of the tumors was useful, although more than half of SD was Ef.1b-2.RECIST criteria corresponded almost perfectly with the WHO criteria, suggesting that RECIST is accurate and useful.

The purpose of this study was to assess the benefits of computed tomography(CT)-guided marking using dyes in patients undergoing lung resection for peripheral nodules.Between January 1997 and August 2002, the location of small pulmonary nodules was identified with the aid of CT-guided marking in 52 patients scheduled for surgery. Dye-injection was performed in 52 patients (indigo carmine, n = 15; indocyanine green, n = 37). The average nodule size was 9.2 mm (range, 3 to 18 mm).The procedure using dyes proved to be easy and safe: the dyes were easily injected near the nodules, and no serious complications ensued. Intraoperatively, indocyanine was superior to indigo carmine with respect to visualization of the dyed pleural surface.The preoperative CT-guided injection of indocyanine green proved to be the most useful and safest technique for identifying peripheral pulmonary nodules in patients scheduled for thoracoscopic surgery or minimal thoracotomy.

Intratumoral hypoxia is one of the resistant factors in radiotherapy and chemotherapy for cancer. Although it is detected by 18F-fluoromisonidazole (FMISO) PET, the relationship between intratumoral hypoxia and oxygen metabolism has not been studied. The purpose of this study was to evaluate the intratumoral perfusion and oxygen metabolism in hypoxic regions using the rat xenograft model. Ten male Fischer rats with C6 glioma (body weight = 220 ± 15 g) were investigated with 18F-FMISO PET and steady-state inhalation method of 15O-labelled gases PET. The tumoral blood flow (TBF), tumoral metabolic rate of oxygen (TMRO2), oxygen extraction fraction (OEF), and tumoral blood volume (TBV) were measured under artificial ventilation with 15O–CO2, 15O–O2, and 15O–CO gases. Multiple volumes of interest (1-mm diameter sphere) were placed on the co-registered 18F-FMISO (3 h post injection) and functional 15O-labelled gases PET images. The TBF, TMRO2, OEF, and TBV values were compared among the three groups classified by the 18F-FMISO uptake as follows: group Low (L), less than 1.0; group Medium (M), between 1.0 and 2.0; and group High (H), more than 2.0 in the 18F-FMISO standardized uptake value (SUV). There were moderate negative correlations between 18F-FMISO SUV and TBF (r = −0.56 and p

Background [11C]methionine (MET) has been used to monitor amino acid metabolism in tumors, the pancreas, liver, and myocardium. The aim of the present study was to standardize [11C]MET positron emission tomography (PET) by optimizing the timing of initiation of the scan and applying correction to the plasma concentrations of neutral amino acids (NAAs), where necessary. Methods Sequential whole-body MET PET/computed tomography (CT) was performed in 11 normal adults after they had fasted for at least 4 h. After whole-body CT for attenuation correction and intravenous bolus injection of MET, the subjects were scanned from the parietal to the groin. The scanning was repeated six to seven times. Decay of radioactivity during the PET scan was corrected to the time of initiation of the first scan. The standardized uptake values (SUVs) were evaluated in various organs by setting regions of interest on the tomographic images. Plasma concentrations of NAAs were examined in relation to the SUV values. Results The SUVs in the pancreas reached their plateau from 6.5 to 11 min after the MET injection, and in the brain, lung, and myocardium, they reached their plateau from 19.6 to 24.1 min. The MET uptake in the spleen and kidney peaked early after the injection and steadily decreased thereafter. The SUVs in the liver and stomach wall rapidly increased during the first 0 to 4.5 min and gradually elevated thereafter during the scan period. Urinary radioactivity in the bladder reached its plateau from 26.1 to 30.6 min after the MET injection. There were no correlations between the plasma concentrations of NAAs and the maximal SUV in any organs. Conclusions The present study revealed the times taken to reach the plateau of MET uptake in various important organs, and little effects of the plasma neutral amino acid concentrations on the SUVs in PET studies conducted after the patients had fasted for at least 4 h. In the MET PET study, 4 h fasting period before MET administration and the scan initiation 20 min after MET administration provide the SUV values independent of scan initiation time and the plasma neutral amino acid concentrations.