Kinetics of a putative hypoxic tracer, 99mTc-HL91, in normoxic, hypoxic, ischemic, and stunned myocardium.

UNLABELLED: 99mTc-4,9-diaza-3,3,10,10-tetramethyldodecan-2,11-dione dioxime (HL91) was developed as a putative hypoxic reagent. This study focused on the myocardial kinetics of 99mTc-HL91 in various oxygen levels and perfusion states. METHODS: The time-activity curve of 99mTc-HL91 was measured in isolated perfused rat heart after the bolus infusion. RESULTS: 99mTc-HL91 was cleared quickly from normoxic hearts, and retention at 30 min after injection was 0.18 +/- 0.02 percentage injected dose per gram of wet weight (mean +/- SE; n = 6). When the concentration of oxygen bubbling through the perfusate was reduced from 100% to 50%, 20%, 5%, and 0%, retention of 99mTc-HL91 increased to 0.47 +/- 0.03 (n = 5), 0.48 +/- 0.03 (n = 5), 0.71 +/- 0.01 (n = 5), and 0.70 +/- 0.02 (n = 5), respectively (P < 0.05). Compartment analysis revealed that the trapping mechanism, which was dependent on tissue oxygen concentration, determined the retention rate. Although not retained in stunned myocardium (0.17 +/- 0.02, n = 5; P = not significant), 99mTc-HL91 was significantly retained when injected before ischemia (1.06 +/- 0.06, n = 5; P < 0.05). CONCLUSION: These results indicate that retention of 99mTc-HL91 correlates well with oxygen level in the perfusate, suggesting that the agent may be a useful marker of the severity of myocardial hypoxia.

[Development of 18F-FDG ([F-18]-2-fluoro-2-deoxy-D-glucose) injection for imaging of tumor reflecting glucose metabolism--results of preclinical studies].

Fluorine-18-2-fluoro-2-deoxy-D-glucose (18F-FDG) injection was prepared by a modification of a method originally developed by Hamacher et al. The dosage form is the injectable solution (2 ml) containing 185 MBq of 18F-FDG at a calibration time. Preclinical studies of the agent were performed. Its radiochemical purity is more than 95% and expiration time is 4 hours after the calibration time at ambient temperature. No toxicity was observed with up to 200 mg/kg and 100 mg/kg of non-radioactive FDG intravenously injected to rats and dogs in single-dose toxicity tests, respectively. Biodistribution studies demonstrated that the radioactivity was mainly distributed into brain (3.0 to 3.3% I.D./Organ at 30 minutes) and heart (4.2 to 5.8% I.D./Organ at 1 to 3 hours) after intravenous injection of the agent to normal rats. In a tumor transplanted mouse model (colon 26), tumor uptake was 10.9 +/- 3.5% I.D./g at 1 hr after intravenous injection of the agent, the radioactivity was retained until 3 hours. The radiation absorbed dose was estimated according to the MIRD Pamphlet based on the biodistribution data both in humans reported by Mejia et al. and rats described in this report. The radiation absorbed dose was not higher than those of commercially available radiopharmaceuticals. In conclusion, the 18F-FDG injection is expected to be useful for further clinical application.

Direct labeling of macroaggregated albumin with indium-111-chloride using acetate buffer.

UNLABELLED: Indium-111-labeled macroaggregated albumin (MAA) would be suitable for combined pulmonary perfusion and ventilation scan using a 99mTc ventilation agent. METHODS: MAA suspended in 0.1 M sodium acetate buffer, pH 5.8, was incubated with 111In-chloride for 30 min at room temperature. An in vitro study of the obtained 111In-MAA was performed for labeling efficiency and stability in human normal serum. The 111In-MAA was intravenously injected into normal mice, and the biodistribution was studied at 15 and 180 min postinjection. A gamma camera image was obtained at 15 min after injection. RESULTS: MAA was directly and stably labeled with 111In-chloride, and the labeling efficiency of the preparation was more than 96%. More than 90% of the administered 111In-MAA was caught in the murine lung. The scintigraphy with 111In-MAA showed a clearly visualized murine lung. CONCLUSION: Indium-111-MAA can be conveniently prepared by direct labeling at room temperature. It provides an alternative perfusion tracer for combined perfusion-ventilation imaging.

Pharmacokinetics of radioiodinated fatty acid myocardial imaging agents in animal models and human studies.

Since the oxidation of long chain fatty acids is the major pathway for energy production for the normoxic myocardium, the use of radiolabeled fatty acids for myocardial imaging continues to be a major area of both basic and clinical research. This paper focuses on a discussion of the kinetics of myocardial uptake of radioiodinated fatty acids, including planar and SPECT imaging of various iodine-123-labeled analogues, and data from animal and isolated heart studies, and where possible, comparison with results of clinical studies. Key examples include iodoalkyl-substituted straight chain fatty acids such as 17-IHDA (17-iodoheptadecanoic acid). These analogues are rapidly metabolized in the myocardium, resulting in release of free radioiodide, and can only be practically used for planar imaging. Terminal iodophenyl-substituted fatty acids illustrate a successful approach of stabilizing radioiodine to overcome the release of free iodide encountered with the straight-chain analogues. These analogues, exemplified by p-IPPA [15-(p-iodophenyl)pentadecanoic acid], are widely used in clinical practice. Although washout can be delayed by increase in the arterial lactate levels by mild exercise, SPECT imaging must still be carefully timed. In contrast to these examples, the ortho iodide-substituted IPPA isomer (ortho- instead of para-phenyl substitution of radioiodide) is a unique example which shows rapid myocardial washout in laboratory animals but nearly irreversible retention in humans. Introduction of methyl-branching is a major important approach which has been successfully used to alter tracer kinetics of radioiodinated fatty acids by increasing myocardial retention. A key example in this class of compounds is 3-(R,S)-BMIPP [15-(p-iodophenyl)-3-(R,S)-methylpentadecanoic acid], an analogue of p-IPPA in which methyl-branching has been introduced into the beta-position of the carbon chain. Although tracer washout is significantly delayed with this structural perturbation, a large number of clinical studies have shown that slow myocardial washout is still observed. Detailed biochemical studies with radioiodinated 3-BMIPP have demonstrated that initial alpha-oxidation produces a metabolite that can then be catabolized by alpha-oxidation. An unexpected and important observation with the (123I]-3-(R,S)-BMIPP agent has been the mis-match between perfusion tracer distribution and the regional BMIPP distribution which has been widely observed in jeopardized, but viable myocardial regions. Another example in the methyl-branched series is DMIPP [15-(p-iodophenyl)- 3,3-dimethylpentadecanoic acid], which has very prolonged myocardial retention and slow washout kinetics although only animal studies have been reported with this agent. Still another more recent approach has been the synthesis and laboratory animal and human evaluation of analogues containing a phenylene bridge in the fatty acid chain. One example is 3-10 [13-(4'-iodophenyl)]-3-(p-phenylene)tridecanoic acid (PHIPA 3-10), which has also proven successful in delaying myocardial tracer washout. This paper focuses on a discussion of the effects of molecular structure on the myocardial uptake and release of these various radioiodinated fatty acid analogues.

Indirect labeling of macroaggregated albumin with indium-111 via diethylenetriaminepentaacetic acid.

It is ideal to perform a simultaneous pulmonary perfusion and ventilation scan in cases of suspected pulmonary thromboembolism. Indium-111 (111In)-diethylenetriaminepentaacetic acid (DTPA)-macroaggregated albumin (MAA) was designed for this purpose. MAA was conjugated with DTPA at a molar ratio of 1:100 and incubated with 111In-chloride for 30 min at room temperature. DTPA-MAA could be labelled with 111In above a 96% labelling efficiency without MAA particle aggregates making their particles larger than desirable. The obtained 111In-DTPA-MAA was intravenously injected into normal mice and their biodistribution was studied at 15 and 180 min after injection. A gamma camera image was obtained 15 min after injection. 111In-DTPA-MAA was stable in vitro and in vivo, and gave high uptake of murine lung in the biodistribution study and clearly visualized murine lung in the scintigraph. Using 111In-DTPA-MAA as a pulmonary perfusion agent, a simultaneous pulmonary perfusion and ventilation scan with technetium-99m-ventilation agents is able to be performed using the dual-isotope technique. 111In-DTPA-MAA may be a potential pulmonary perfusion agent.

Metabolism of iodine-123-BMIPP in perfused rat hearts.

UNLABELLED: Increased clinical use of 123I-labeled 15-(p-iodophenyl)-3-(R,S)-methyl- pentadecanoic acid ([123I]BMIPP) revealed discordance between BMIPP uptake and that of perfusion agents, which was inexplicable due to the uncertainty of its myocardial metabolism. This study clarifies the metabolic fate of BMIPP and its relation to substrates in isolated rat hearts. METHODS: Rat hearts were perfused with 5 mmole/liter HEPES buffer containing various energy substrates and 1% bovine serum albumin. The buffer was recirculated for 4 hr after bolus injection of [123I]BMIPP. Heart time-activity curves were monitored externally. After perfusion, the radioactivity in the heart and recirculated buffer was measured. The metabolites in the buffer were then extracted and analyzed by HPLC and TLC. RESULTS: when 0.4 mmole/liter oleate was the energy substrate, more than eight radioactive BMIPP metabolites were detected. The metabolites in the coronary effluent depended on the energy substrate in the buffer. The radioactivity in the heart at the end of the perfusion period was significantly higher when 0.4 mmole/liter oleate (28.0% +/- 1.2% ID/g, mean +/- s.e.m.) or 10 mmole/liter glucose with 25 U/liter insulin (43.9% +/- 2.2% ID/g) were the substrates compared to when 5 mmole/liter acetate (8.5% +/- 0.4% ID/g) or 0.4 mmole/liter cold BMIPP (6.2% +/- 0.3% ID/g) were the substrates. The distribution of metabolites suggests that oleate stimulated both alpha and beta oxidations, whereas glucose with insulin inhibited both. Acetate also stimulated alpha oxidation but not beta oxidation. Cold BMIPP strongly inhibited both alpha- and beta-oxidations, and little alpha oxidation occurred compared to beta-oxidation. CONCLUSION: These results suggest that [123I]BMIPP is metabolized in the myocardium and the metabolism is closely related to myocardial carbohydrate utilization.

Intestinal absorption of pyridoxal 5'-phosphate at physiological levels in rats.

The intestinal absorption of pyridoxal 5'-phosphate (PLP) at physiological levels (10(-7) -10(-6) M) was studied in comparison with that of pyridoxal (PL) in rat, using in vitro everted sac and an intestinal preparation that permitted continuous in situ collection of mesenteric venous blood. After PLP administration (10(-6) -10(-3) M) in situ, larger amounts of PLP were found in the mesenteric venous plasma than after PL administration at the same dose. The amount of PLP found in the mesenteric venous plasma was dependent on its dose at lower concentrations up to 10(-4) M but became independent at higher concentrations. After PL administration at various doses, the amount of PL found in the mesenteric venous blood increased linearly with the dose. When various concentrations of PLP were added to the mucosal side, under the in vitro condition with protection from alkaline phosphate hydrolysis, PLP was detected in the serosal side and the extent of PLP transport was dependent on the initial concentration of PLP in the mucosal side. When various concentrations of PL were added to the mucosal side, the extent of PL transport was independent of the initial concentration of PL in the mucosal side. In rat pretreated with actinomycin D, PLP transport in vitro was inhibited but not that of PL. N2-induced anoxia and pyridoxamine 5'-phosphate and anion transport inhibitor (4,4'-diisothiocyanostilben-2,2'-disulfonic acid disodium salt) showed no effect on PLP transport. These results suggest that PLP can be absorbed in the phosphorylated form and imply the presence of a saturable process for direct absorption of PLP itself and a diffusive process for PL absorption. In addition, the result of the in vivo neonatal experiment suggests that the neonatal intestine also can transport PLP in phosphorylated form.