In vivo imaging of microglial activation using a peripheral benzodiazepine receptor ligand: [11C]PK-11195 and animal PET following ethanol injury in rat striatum

OBJECTIVE: To investigate whether [(11)C]PK-11195, a specific peripheral benzodiazepine receptors (PBRs) ligand for positron emission tomography (PET), can show activated microglia in a rat brain injury model. METHODS: On day 1, ethanol was injected into the rat's right striatum (ST) using a stereotaxic operative procedure. On day 3, head magnetic resonance imaging (MRI) scans for surgically treated rats were performed to evaluate ethanol injury morphologically. On day 4, dynamic PET scans (17 injured rats and 7 non-injured controls) were performed for 60 min with an animal PET scanner under chloral hydrate anesthesia following a bolus injection of [(11)C]PK-11195 through tail vein. Because PBRs are present throughout the brain, there is no suitable receptor-free reference region. The reference tissue model may not be applicable because of low target to background ratio for low affinity of [(11)C]PK-11195 to PBRs. We evaluated the PBRs binding with regions of interest (ROIs)-based approach to estimate total distribution volume (V). We used an integral from 0 min to 60 min (V (60)) as an estimate of V. On the coronal PET image, ROIs were placed on bilateral ST. Differences in right/left ST V (60) ratios between lesioned and unlesioned control rats were compared using unpaired t tests. Immunohistochemical staining was performed for confirming the presence of activated microglia following decapitation on the PET experiment day. RESULTS: The right/left ST V (60) ratios in lesioned rats (1.07 +/- 0.08) were significantly higher than those in unlesioned control rats (1.00 +/- 0.06, P < 0.05). On immunohistochemical staining, activated microglia were exclusively observed in the injured right ST but not in the noninjured left ST of the injury rats and the bilateral ST of the non-injured control rats. CONCLUSIONS: These results suggest that [(11)C]PK-11195 PET imaging would be a useful tool for evaluating microglial activation in a rat brain injury model.     

1H‐[13C] NMR spectroscopy of the rat brain during infusion of [2‐13C] acetate at 14.1 T

Full signal intensity ¹H‐[¹³C] NMR spectroscopy, combining a preceding ¹³C‐editing block based on an inversion BISEP (B₁‐insensitive spectral editing pulse) with a spin‐echo coherence–based localization, was developed and implemented at 14.1 T. ¹³C editing of the proposed scheme was achieved by turning on and off the ¹³C adiabatic full passage in the ¹³C‐editing block to prepare inverted and noninverted ¹³C‐coupled ¹H coherences along the longitudinal axis prior to localization. The novel ¹H‐[¹³C] NMR approach was applied in vivo under infusion of the glia‐specific substrate [2‐¹³C] acetate. Besides a ～50% improvement in sensitivity, spectral dispersion was enhanced at 14.1 T, especially for J‐coupled metabolites such as glutamate and glutamine. A more distinct spectral structure at 1.9–2.2 ppm(parts per million) was observed, e.g., glutamate C3 showed a doublet pattern in both simulated ¹H spectrum and in vivo ¹³C‐edited ¹H NMR spectra. Besides ¹³C time courses of glutamate C4 and glutamine C4, the time courses of glutamate C3 and glutamine C3 obtained by ¹H‐[¹³C] NMR spectroscopy were reported for the first time. Such capability should greatly improve the ability to study neuron‐glial metabolism using ¹H‐observed ¹³C‐edited NMR spectroscopy. Magn Reson Med, 2010. © 2010 Wiley‐Liss, Inc.     

De-coupling of blood flow and metabolism in the rat brain induced by glutamate

Objective  Glutamate plays an essential role in neuronal cell death in many neurological disorders. In this study, we examined both glucose metabolism and cerebral blood flow in the same rat following infusion of glutamate or ibotenic acid using the dual-tracer technique. The effects of MK-801, an NMDA receptor antagonist, and NBQX, an AMPA-kainate receptor antagonist, on the changes in the glucose metabolism and cerebral blood flow induced by glutamate were also examined. Methods  The rats were microinjected with glutamate (1 μmol/μl, 2 μl) or ibotenic acid (10 μg/μl, 1 μl) into the right striatum, and dual-tracer autoradiograms of [¹⁸F]FDG and [¹⁴C]IMP were obtained. MK-801 and NBQX were injected intravenously about 45 and 30 min, respectively, after the infusion of glutamate. Results  De-coupling of blood flow and metabolism was noted in the glutamate-infused hemisphere (as assessed by no alteration of [¹⁸F]FDG uptake and significant decrease of [¹⁴C]IMP uptake). Pretreatments with MK-801, NBQX, or combined use of MK-801 and NBQX did not affect the de-coupling of the blood flow and metabolism induced by glutamate. A histochemical study revealed that about 20% neuronal cell death had occurred in the striatum at 105 min after the infusion of glutamate. In addition, a significant increase of the [¹⁸F]FDG uptake and decrease of [¹⁴C]IMP uptake were also seen in the rat brain infused with ibotenic acid. Conclusion  These results indicate that glutamate and ibotenic acid caused a significant de-coupling of blood flow and glucose metabolism in the intact rat brain during the early phase of neurodegeneration. It is necessary to evaluate the relation between metabotropic glutamate receptors and de-coupling of blood flow and metabolism.     

Evaluation of [14C]phenylacetate as a prototype tracer for the measurement of glial metabolism in the rat brain

[¹⁴C]Phenylacetate was designed as a prototype tracer for the measurement of glial metabolism, and its potency in comparison with that of [¹⁴C]acetate was evaluated in this study. Normal rats were intravenously injected with [¹⁴C]phenylacetate or [¹⁴C]acetate, and radioactivity concentrations were measured in the plasma, cerebral cortex, cerebellum and peripheral tissues by dissection method. In addition, [¹⁴C]phenylacetate uptake in the rat brain was compared by autoradiography with that of [¹⁴C]acetate following the injection of fluorocitrate, a selective glial toxin, into the brain. [¹⁴C]Phenylacetate was rapidly taken up into the brain and was retained at high levels up to 20 min postadministration. The levels of [¹⁴C]phenylacetate in the cerebral cortex were about threefold higher than those of [¹⁴C]acetate at 1 min postinjection. Microinjection of fluorocitrate into the right striatum resulted in a significant decrease of the uptake of both [¹⁴C]phenylacetate and [¹⁴C]acetate into the right striatum. Radiochemical analysis confirmed the rapid hydrolysis of [¹⁴C]phenylacetate in the rat brain, with less than 20% of radioactivity representing unmetabolized [¹⁴C]phenylacetate at 1 min postinjection. These results suggest that [¹⁴C]phenylacetate is rapidly taken up into the brain and is hydrolyzed and converted to [¹⁴C]acetate. [¹⁴C]Phenylacetate may have the potential to serve as a tracer for the measurement of glial metabolism in an intact brain.     

Comparison of [<Superscript>18</Superscript>F]FHBG and [<Superscript>14</Superscript>C]FIAU for imaging of <Emphasis Type="Italic">HSV1-tk</Emphasis> reporter gene expression: adenoviral infection vs stable transfection

Earlier studies involving comparison of different reporter probes have shown conflicting results between pyrimidine nucleosides [e.g., 2'-fluoro-2'-deoxy-1--d-arabinofuranosyl-5-iodouracil (FIAU)] and acycloguanosine derivatives [e.g., penciclovir (PCV), 9-(4-fluoro-3-hydroxymethylbutyl)guanine (FHBG)]. We hypothesized that this reported discrepancy may be related to how the reporter gene is delivered to the cells—stably transfected vs adenoviral infection. We directly compared the uptake characteristics of [¹⁸F]FHBG, [³H]PCV, and [¹⁴C]FIAU in cell culture and in vivo using an adenoviral mediated gene transfer model and stably transfected cells. We further compared the uptake of three reporter probes using both HSV1-tk and a mutant HSV1-sr39tk expressing cells to assess the optimal reporter probe/reporter gene combination. [¹⁴C]FIAU accumulation was greater than that of [³H]PCV and [¹⁸F]FHBG in control cells and in HSV1-tk stably transfected cells (P<0.001). After infection of C6 cells with AdCMV-HSV1-tk (1.5×10⁸ pfu), [¹⁸F]FHBG and [³H]PCV accumulation was significantly greater than that of [¹⁴C]FIAU (P<0.01). [¹⁸F]FHBG and [³H]PCV accumulated to a significantly greater extent than [¹⁴C]FIAU in C6-stb-sr39tk+ and AdCMV-HSV1-sr39tk infected C6 cells (P<0.001). Results from the nude mice supported the results in cell culture. [¹⁴C]FIAU led to significantly higher %ID/g in C6-stb-tk+ xenografts than [¹⁸F]FHBG (P<0.05); however, compared with [¹⁴C]FIAU, [¹⁸F]FHBG led to as high %ID/g in HSV1-tk expressing hepatocytes and to significantly greater %ID/g in C6-stb-sr39tk+ xenografts and HSV1-sr39tk expressing hepatocytes. Dynamic sequential images showed that [¹⁸F]FHBG was well retained in HSV1-sr39tk expressing cells (C6-stb-sr39tk+) for at least 4 h after injection, while it was rapidly cleared from HSV1-tk expressing cells (MH3924A-stb-tk+). [¹⁴C]FIAU accumulated in HSV1-tk stably expressing cells to a greater extent than either [³H]PCV or [¹⁸F]FHBG. However, the accumulation of [³H]PCV and [¹⁸F]FHBG in adenoviral infected C6 cells or hepatocytes was equivalent to or greater than that of [¹⁴C]FIAU. These results may be due to intracellular biochemical changes (e.g., thymidine) when cells are infected with adenovirus. For adenoviral studies, the [¹⁸F]FHBG/HSV1-sr39tk combination was shown to be more sensitive than the [¹⁴C]FIAU/HSV1-tk combination HSV1-tk.     

NanoPET imaging of [18F]fluoromisonidazole uptake in experimental mouse tumours

Purpose The purpose of this study was to assess the potential and utility of ultra-high-resolution hypoxia imaging in various murine tumour models using the established hypoxia PET tracer [¹⁸F]fluoromisonidazole ([¹⁸F]FMISO).Methods [¹⁸F]FMISO PET imaging was performed with the dedicated small-animal PET scanner NanoPET (Oxford Positron Systems) and ten different human tumour xenografts in nude mice as well as B16 melanoma tumours in syngeneic Balb/c mice. For comparison, [¹⁸F]fluorodeoxyglucose ([¹⁸F]FDG) PET scans were also performed in the mice bearing human tumour xenografts.Results In 10 out of 11 experimental tumour models, [¹⁸F]FMISO PET imaging allowed clear-cut visualisation of the tumours. Inter- and intratumoural heterogeneity of tracer uptake was evident. In addition to average TMRR (tumour-to-muscle retention ratio including all voxels in a volume of interest (VOI)), the parameters TMRR_75% and TMRR₅ (tumour-to-muscle retention ratio including voxels of 75% or more of the maximum radioactivity in a VOI and the five hottest pixels, respectively) also served as measures for quantifying the heterogeneous [¹⁸F]FMISO uptake in the tumours. The variability observed in [¹⁸F]FMISO uptake was related neither to tumour size nor to the injected mass of the radiotracer. The pattern of normoxic and hypoxic regions within the human tumour xenografts, however, correlated with glucose metabolism as revealed by comparison of [¹⁸F]FDG and [¹⁸F]FMISO images.Conclusion This study demonstrates the feasibility and utility of [¹⁸F]FMISO for imaging murine tumour models using NanoPET.     

Development of a modular system for the synthesis of PET [C]labelled radiopharmaceuticals

[¹¹C]labelled radiopharmaceuticals as N-[¹¹C]methyl-choline ([¹¹C]choline), l-(S-methyl-[¹¹C])methionine ([¹¹C]methionine) and [¹¹C]acetate have gained increasing importance in clinical PET and for the routine production of these radiopharmaceuticals, simple and reliable modules are needed to produce clinically relevant radioactivity. On the other hand, flexible devices are needed not only for the routine synthesis but also for more complex applications as the development of new tracers. The aim of this work was the adaptation of an Eckert Ziegler modular system for easy routine synthesis of [¹¹C]choline, [¹¹C]methionine and [¹¹C]acetate using components that account for straightforward scaling up and upgrades.     

A simplified autoradiographic method with alpha-[14C]methyl-tryptophan to measure serotonin synthesis rate in the rat brain

BACKGROUND: To minimize blood sampling necessary for the autoradiographic (ARG) method using alpha-[(14)C]methyl-tryptophan (alpha-MTrp), a simplified method using a standard exposure time (theta(S)) was proposed and the accuracy of the method was evaluated. METHODS: A total of 168 rats from three sets of experiments with different serotonergic drugs to evaluate changes in cerebral serotonin (5-hydroxytryptamine; 5-HT) synthesis rate were used. In the acute treatment study, rats received a drug injection 30 min before the tracer experiment. In the repeated treatment study, the same dose of drug was injected for 7 days. The two-time point method provided a parameter of brain trapping constant (K*) from the slope of linear regression of the Patlak plot, from which 5-HT synthesis rate is calculated. The measured exposure time (theta(M)) for 60 and 150 min after the tracer injection obtained from individual blood sampling in the original method and the theta(S) calculated from theta(M) were used for evaluation of differences in K* values. RESULTS: No significant difference in theta(M) was noted among different experiments, although plasma radioactivity at the end of experiment was significantly different between the acute and the repeated treatments for one of the three drugs. No difference in K* for each treatment was observed between the original method and the simplified single blood sampling method because there was no difference in theta(M) among three experiments and between theta(M) and theta(S). CONCLUSION: The simplified alpha-MTrp ARG method, which uses theta(S) and a single arterial blood sample at the end of each experiment, can be used for the measurement of 5-HT synthesis rate in the rat brain.     

Changes in myocardial oxidative metabolism after biventricular pacing as evaluated by [11C]acetate positron emission tomography

A 70-year-old woman with dilated cardiomyopathy and recurrent severe heart failure was admitted for biventricular pacing (BVP), which was recently reported to have clinical efficacy for severe heart failure with intraventricular conduction delay. An electrocardiogram showed complete left bundle branch block, and the QRS interval was markedly prolonged at 195 msec. Echocardiogram showed marked dilatation, diffuse hypokinesis and dyssynchrony of the left ventricle, and grade III mitral valve regurgitation. The patient underwent implantation of an atriobiventricular pacemaker and three pacing leads transvenously. The QRS interval shortened to 165 msec immediately after the BVP therapy, and improvements in echocardiographic parameters were seen at 5 months after BVP therapy. Myocardial oxidative metabolism was assessed by the monoexponential clearance rate of [¹¹C]acetate (Kmono) as measured by positron emission tomography (PET), and myocardial efficiency was assessed by the work metabolic index (WMI) at 1 and 5 months after the BVP therapy. The PET images obtained 5 months after BVP therapy showed a decrease in the clearance of [¹¹C]acetate compared with that obtained 1 month after BVP therapy. The Kmono of the whole left ventricle decreased from 0.051 at 1 month to 0.038 min⁻¹ at 5 months after BVP therapy, and that of the septum, anterior wall, lateral wall and posterior wall also decreased. The WMI increased from 4.2×10⁶ to 6.8×10⁶ mmHg·ml/m². These results suggest that BVP improved left ventricular function without increasing myocardial oxidative metabolism, resulting in improved myocardial efficiency, and that BVP may improve the long-term prognosis of heart failure patients with ventricular dyssynchrony. [¹¹C]acetate PET is a useful method of evaluating global and regional myocardial oxidative metabolism in patients who have undergone BVP therapy.     

Identification and regional distribution in rat brain of radiometabolites of the dopamine transporter PET radioligand [<Superscript>11</Superscript>C]PE2I

Purpose We aimed to determine the composition of radioactivity in rat brain after intravenous administration of the dopamine transporter radioligand, [¹¹C]PE2I. Methods PET time-activity curves (TACs) and regional brain distribution ex vivo were measured using no-carrier-added [¹¹C]PE2I. Carrier-added [¹¹C]PE2I was administered to identify metabolites with high-performance liquid radiochromatography (RC) or RC with mass spectrometry (LC-MS and MS-MS). The stability of [¹¹C]PE2I was assessed in rat brain homogenates. Results After peak brain uptake of no-carrier-added [¹¹C]PE2I, there was differential washout rate from striata and cerebellum. Thirty minutes after injection, [¹¹C]PE2I represented 10.9 ± 2.9% of the radioactivity in plasma, 67.1 ± 11.0% in cerebellum, and 92.5 ± 3.2% in striata, and was accompanied by two less lipophilic radiometabolites. [¹¹C]PE2I was stable in rat brain homogenate for at least 1 h at 37°C. LC-MS identified hydroxylated PE2I (1) (m/z 442) and carboxyl-desmethyl-PE2I (2) (m/z 456) in brain. MS-MS of 1 gave an m/z 442→424 transition due to H₂O elimination, so verifying the presence of a benzyl alcohol group. Metabolite 2 was the benzoic acid derivative. Ratios of ex vivo measurements of [¹¹C]PE2I, [¹¹C]1, and [¹¹C]2 in striata to their cognates in cerebellum were 6.1 ± 3.4, 3.7 ± 2.2 and 1.33 ± 0.38, respectively, showing binding selectivity of metabolite [¹¹C]1 to striata. Conclusion Radiometabolites [¹¹C]1 and [¹¹C]2 were characterized as the 4-hydroxymethyl and 4-carboxyl analogs of [¹¹C]PE2I, respectively. The presence of the pharmacologically active [¹¹C]1 and the inactive [¹¹C]2 is a serious impediment to successful biomathematical analysis.     

Measurement of myocardial fatty acid esterification using [1-11C]palmitate and PET: comparison with direct measurements of myocardial triglyceride synthesis

Background  The purpose of the present study was to assess the accuracy of rates of myocardial fatty acid esterification (MFAE) obtained using positron emission tomography (PET). Methods and results  Sixteen dogs were studied after an overnight fast (FAST), during a euglycemic hyperinsulinemic clamp (CLAMP), or during infusion of intralipid (IL) or IL plus dobutamine (IL/DOB). MFAE was quantified using [1-¹¹C]palmitate and PET and compared to the rate of triglyceride (TG) synthesis measured using [1-¹³C]palmitate and tissue sampling. Plasma free fatty acid (FFA) concentration varied ~20-fold across groups, with this variation in FFA availability accompanied by a ~20-fold range in TG synthesis. MFAE varied ~12-fold across groups, and was significantly correlated with TG synthesis (R = 0.80, P < .001). MFAE, however, was 3- to 4-fold higher than TG synthesis in FAST, CLAMP, and IL, but only ~50% higher when cardiac work was increased in IL/DOB, suggesting that MFAE reflects, in part, the incorporation of label into amino acids via TCA cycle exchange reactions. Conclusions  Changes in MFAE parallel changes in TG synthesis, at least in the basal state. Although the data need to be interpreted cautiously, such measurements should be useful for quantifying acute changes in FFA storage by the heart in various pathophysiological states.     

Two routes to [C-carbonyl]organo-isocyanates utilizing [C]phosgene ([C]organo-isocyanates from [C]phosgene)

Two generic radiosynthetic routes for the preparation of [¹¹C-carbonyl]isocyanates have been developed. Reaction of N-organo-sulfinylamines; RNSO, (R = Me, Et, allyl, cyclohexyl and phenyl) with [¹¹C]phosgene gave the corresponding [¹¹C-carbonyl]isocyanates in good radiochemical yield (53–68%) from [¹¹C]phosgene (decay corrected) in ca 16 min from EOB. Alternatively, the reaction of [¹¹C]phosgene with N,N′-organo-ureas; (RNH)₂CO, (R = Me, Et, Pr and phenyl) also gave the corresponding [¹¹C-carbonyl]isocyanates in moderate radiochemical yield (9–37%) from [¹¹C]phosgene (decay corrected) in ca 16 min from EOB. For identification, the [¹¹C-carbonyl]organo-isocyanates were derivatized with 1-(2-methoxyphenyl)piperazine in situ to [¹¹C-carbonyl]carboxamides and the position of radiolabelling in the carbonyl group confirmed by [^11/13C]co-labeling and subsequent carbon-13 NMR spectroscopy.     

An automated SPE-based high-yield synthesis of [11C]acetate and [11C]palmitate: no liquid-liquid extraction, solvent evaporation or distillation required

Introduction

An automated method is described for the rapid and high-yield synthesis of two of the most commonly used radioactive fatty acids: [¹¹C]acetate and [¹¹C]palmitate.

Methods

Reaction of [¹¹C]CO₂ with the respective Grignard reagents in diethyl ether solution proceeded for 2 min at 40°C. Quenching of the reaction and liberation of nonreacted [¹¹C]CO₂ occurred upon addition of a fourfold molar excess of aqueous 0.1 M HCl (acetate) or nonaqueous HCl/Et₂O (palmitate). Labeled products were then purified by adsorption to an Alumina-N Sep-Pak Plus cartridge and eluted with either aqueous NaH₂PO₄ solution (acetate) or 100% ethanol (palmitate).

Results

High-performance liquid chromatography analysis confirmed that the radiochemical purity of each product was >98%, and decay-corrected radiochemical yields averaged 33% for [¹¹C]palmitate and 40% for [¹¹C]acetate.

Conclusion

The method requires no liquid–liquid extraction, solvent evaporation or distillation capabilities and can be readily adapted to existing radiosynthesis modules.     

[1-<Superscript>11</Superscript>C]Acetate uptake is not increased in renal cell carcinoma

Purpose The purpose of this study was to investigate the potential of [1-¹¹C]acetate (AC) as a metabolic tracer for renal cell cancer in human subjects. Methods Twenty-one patients with suspected kidney tumours were investigated with AC and dynamic PET. AC uptake was scored on a five-step scale. Tumour localisation was known from CT/MRI. Histology was available in 18/21 patients. The results in these 18 patients are reported. Results AC uptake by the tumour was less than (n = 11), equal to (n = 5) or higher than (n = 2) uptake in the surrounding renal parenchyma. Histological tumour types showed a typical distribution, with a predominance of clear cell carcinomas (n = 14) and only a small number of papillary cell carcinomas (n = 2) and oncocytomas (n = 2). Only the benign oncocytomas were highly positive with AC. Conclusion In most kidney tumours the AC accumulation was not higher than in normal kidney parenchyma. Therefore, AC PET cannot be recommend for the characterisation of a renal mass.     

Biodistribution and radiation dosimetry of [11C]raclopride in healthy volunteers

Purpose This study reports on the whole-body biodistribution and radiation dosimetry of [¹¹C]raclopride, a dopamine D₂ receptor antagonist.Methods In three healthy male volunteers, whole-body scans were performed up to 2 h following i.v. injection of 320±65 MBq [¹¹C]raclopride. Transmission scans (3 min per step, eight or nine steps according to the height of the subject) in 2D mode were used for subsequent attenuation correction of emission scans. Emission scans (1 min per step, eight or nine steps) were acquired over 2 h. Venous blood samples and urine were collected up to 2 h after injection of the radiotracer. For each subject, the percentage of injected activity measured in regions of interest over brain, intestine, lungs, kidneys and liver was fitted to a mono-exponential model, as an uptake phase followed by a mono-exponential washout, for urinary bladder to generate time–activity curves. Using the MIRD method, several source organs were considered in estimating residence time and mean effective radiation absorbed doses.Results Blood pressure and ECG findings remained unchanged after tracer injection. The analysed blood and urine pharmacological parameters did not change significantly after [¹¹C]raclopride injection. The primary routes of clearance were renal and intestinal. Ten minutes after injection, high activities were observed in the gall-bladder, kidneys and liver. High activity was observed in the gall-bladder during the whole study. The kidneys, urinary bladder wall, liver and gall-bladder received the highest absorbed doses. The average effective dose of [¹¹C]raclopride was estimated to be 6.7±0.4 Sv/MBq.Conclusion The amount of [¹¹C]raclopride required for adequate dopamine D₂ receptor imaging results in an acceptable effective dose equivalent, permitting two or three repeated clinical PET imaging studies, with the injection of 222 MBq for each study.     

[C]Formaldehyde revisited: considerable concurrent [C]formic acid formation in the low-temperature conversion of [C]carbon dioxide into [C]formaldehyde

The reduction of [¹¹C]carbon dioxide with lithium aluminium hydride in diethyl ether at temperatures ranging from −56°C to 19°C was studied. In contrast to what others have reported, considerable amounts of [¹¹C]formic acid were found at all studied temperatures.     

Difference of 14C turnovers in brain and in transplanted glioma after intravenous injection of 14C-1-pyruvate into rats

Carbon 14 from ¹⁴C-1-pyruvate injected intravenously into glioma-transplanted rats was incorporated into various compounds in the brain and in the tumor. In the brain the majority of activity was found in CO₂ (60%), and minor activities were found in alanine, lactate (15%), glutamate, and aspartate, with decreasing order, 5 min after injection. In the tumor, at 5 min, the largest activity was in lactate (56%), and lower activities were found in CO₂ (24%), alanine, glutamate, and aspartate. The total ¹⁴C concentration in the tumor was twice that in the brain at 5 min and 15 min. The result was in accordance with the prediction that in brain, where the mitochondrial function is active, ¹⁴C-1-pyruvate will be oxidized completely into ¹⁴CO₂, and that in tumor, where the mitochondrial function is insufficient, ¹⁴C-1-pyruvate will be converted only into ¹⁴C-lactate and prevent further degradation. It may be assumed that this difference in the turnover of ¹⁴C of ¹⁴C-1-pyruvate between brain and tumor could constitute a basis for the hot visualization of human brain tumor using cyclotron-produced ¹¹C-1-pyruvate and positronemission tomography.     

Use of reference tissue models for quantification of histamine H1 receptors in human brain by using positron emission tomography and [11C]doxepin

The aim of the present study is to evaluate the validity of the simplified reference tissue model (SRTM) and of Logan graphical analysis with reference tissue (LGAR) for quantification of histamine H1 receptors (H1Rs) by using positron emission tomography (PET) with [11C]doxepin. These model-based analytic methods (SRTM and LGAR) are compared to Logan graphical analysis (LGA) and to the one-tissue model (1TM), using complete datasets obtained from 5 healthy volunteers. Since HIR concentration in the cerebellum can be regarded as negligibly small, the cerebellum was selected as the reference tissue in the present study. The comparison of binding potential (BP) values estimated by LGAR and 1TM showed good agreement; on the other hand, SRTM turned out to be unstable concerning parameter estimation in several regions of the brain. By including the results of noise analysis, LGAR became a reliable method for parameter estimation of [11C]doxepin data in the cortical regions.     

[<Superscript>18</Superscript>F]EF3 is not superior to [<Superscript>18</Superscript>F]FMISO for PET-based hypoxia evaluation as measured in a rat rhabdomyosarcoma tumour model

Purpose  The aim of this investigation was to quantitatively compare the novel positron emission tomography (PET) hypoxia marker 2-(2-nitroimidazol-1-yl)-N-(3[¹⁸F],3,3-trifluoropropyl)acetamide ([¹⁸F]EF3) with the reference hypoxia tracer [¹⁸F]fluoromisonidazole ([¹⁸F]FMISO). Methods  [¹⁸F]EF3 or [¹⁸F]FMISO was injected every 2 days into two separate groups of rats bearing syngeneic rhabdomyosarcoma tumours. In vivo PET analysis was done by drawing regions of interest on the images of selected tissues. The resulting activity data were quantified by the percentage of injected radioactivity per gram tissue (%ID/g) and tumour to blood (T/B) ratio. The spatial distribution of radioactivity was defined by autoradiography on frozen tumour sections. Results  The blood clearance of [¹⁸F]EF3 was faster than that of [¹⁸F]FMISO. The clearance of both tracers was slower in tumour tissue compared with other tissues. This results in increasing T/B ratios as a function of time post tracer injection (p.i.). The maximal [¹⁸F]EF3 tumour uptake, compared to the maximum [¹⁸F]FMISO uptake, was significantly lower at 2 h p.i. but reached similar levels at 4 h p.i. The tumour uptake for both tracers was independent of the tumour volume for all investigated time points. Both tracers showed heterogeneous intra-tumoural distribution. Conclusions  [¹⁸F]EF3 tumour uptake reached similar levels at 4 h p.i. compared with tumour retention observed after injection of [¹⁸F]FMISO at 2 h p.i. Although [¹⁸F]EF3 is a promising non-invasive tracer, it is not superior over [¹⁸F]FMISO for the visualisation of tumour hypoxia. No significant differences between [¹⁸F]EF3 and [¹⁸F]FMISO were observed with regard to the intra-tumoural distribution and the extra-tumoural tissue retention.     

Age-related changes of the [11C]CFT binding to the striatal dopamine transporters in the Fischer 344 rats: a PET study

We investigated the age-related changes of the binding of [11C]CFT to striatal dopamine transporters (DATs) in vivo in Fischer 344 rats by positron emission tomography (PET). The tissue dissection method represented an age-related decrease in the uptake ratio of the striatum to the cerebellum and in the specific binding-to-nonspecific binding ratio of [11C]CFT. PET demonstrated an age-dependent decrease in the striatal uptake of [11C]CFT, however, the kinetic analysis represented the age-related decrease in both the association rate constant (k3) and dissociation rate constant (k4), but not the binding potential (k3/k4) that was a parameter including both of density and affinity of the binding sites. The PET finding was not necessarily coincident with the result investigated in vitro previously. Therefore, careful interpretation is necessary for PET studies using [11C]CFT and small animals such as rats.