In vivo assessment of cardiac insulin resistance by nuclear probes using an iodinated tracer of glucose transport.

PURPOSE: Insulin resistance, implying depressed cellular sensitivity to insulin, is a risk factor for type 2 diabetes and cardiovascular disease. This study is the first step towards the development of a technique of insulin resistance measurement in humans with a new tracer of glucose transport, [(123)I]6-deoxy-6-iodo-D-glucose (6DIG). METHODS: We investigated 6DIG kinetics in anaesthetised control rats and in three models of insulin-resistant rats: fructose fed, Zucker and ZDF. The study of myocardial 6DIG activity was performed under two conditions: first, 6DIG was injected under the baseline condition and then it was injected after a bolus injection of insulin. After each injection, radioactivity was measured over 45 min by external detection via NaI probes, in the heart and blood. A tri-compartment model was developed to obtain fractional transfer coefficients of 6DIG from the blood to the heart. RESULTS: These coefficients were significantly increased with insulin in control rats and did not change significantly in insulin-resistant rats. The ratio of the coefficient obtained under insulin to that obtained under basal conditions gave an index of cardiac insulin resistance for each animal. The mean values of these ratios were significantly lower in insulin-resistant than in control rats: 1.16 +/- 0.06 vs 2.28 +/- 0.18 (p < 0.001) for the fructose-fed group, 0.92 +/- 0.05 vs 1.62 +/- 0.25 (p < 0.01) for the Zucker group and 1.34 +/- 0.06 vs 2.01 +/- 0.26 (p < 0.05) for the ZDF group. CONCLUSION: These results show that 6DIG could be a useful tracer to image cardiac insulin resistance.

Assessment of insulin resistance in fructose-fed rats with 125I-6-deoxy-6-iodo-D-glucose, a new tracer of glucose transport.

PURPOSE: Insulin resistance, characterised by an insulin-stimulated glucose transport defect, is an important feature of the pre-diabetic state that has been observed in numerous pathological disorders. The purpose of this study was to assess variations in glucose transport in rats using (125)I-6-deoxy-6-iodo-D-glucose (6DIG), a new tracer of glucose transport proposed as an imaging tool to assess insulin resistance in vivo. METHODS: Two protocols were performed, a hyperinsulinaemic-euglycaemic clamp and a normoinsulinaemic-normoglycaemic protocol, in awake control and insulin-resistant fructose-fed rats. The tracer was injected at steady state, and activity in 11 tissues and the blood was assessed ex vivo at several time points. A multicompartmental mathematical model was developed to obtain fractional transfer coefficients of 6DIG from the blood to the organs. RESULTS: Insulin sensitivity of fructose-fed rats, estimated by the glucose infusion rate, was reduced by 40% compared with control rats. At steady state, 6DIG uptake was significantly stimulated by insulin in insulin-sensitive tissues of control rats (basal versus insulin: diaphragm, p < 0.01; muscle, p<0.05; heart, p<0.001), whereas insulin did not stimulate 6DIG uptake in insulin-resistant fructose-fed rats. Moreover, in these tissues, the fractional transfer coefficients of entrance were significantly increased with insulin in control rats (basal vs insulin: diaphragm, p<0.001; muscle, p<0.001; heart, p<0.01) whereas no significant changes were observed in fructose-fed rats. CONCLUSION: This study sets the stage for the future use of 6DIG as a non-invasive means for the evaluation of insulin resistance by nuclear imaging.

Assessment of insulin sensitivity in vivo in control and diabetic mice with a radioactive tracer of glucose transport: [125I]-6-deoxy-6-iodo-D-glucose.

BACKGROUND: Impairment of insulin-stimulated glucose transport is a characteristic of type 2 diabetes. A radioactive glucose analogue has been synthesized: [(125)I]-6-deoxy-6-iodo-D-glucose. Its biological behaviour in vitro is similar to that of 3-O-methyl-D-glucose, the reference tracer of glucose transport. The aim of the present study was to determine the ability of [(125)I]-6-deoxy-6-iodo-D-glucose to evaluate variations in glucose transport in vivo. METHODS: Biodistributions of [(125)I]-6-deoxy-6-iodo-D-glucose were performed with or without exogenous insulin (iv injection of 1.5 IU/kg) in db/+ non-diabetic control mice and in db/db type 2 diabetic mice, exhibiting a severe insulin resistance characterized by a lack of increase in glucose uptake in response to insulin. RESULTS: In db/+ mice, insulin increased [(125)I]-6-deoxy-6-iodo-D-glucose transport by 30% in most insulin-sensitive tissues (heart, diaphragm and skeletal muscle, p < 0.05) and had no effect in other organs. In db/db mice, [(125)I]-6-deoxy-6-iodo-D-glucose transport in these organs was not modified by insulin. CONCLUSION: [(125)I]-6-deoxy-6-iodo-D-glucose is able to trace in vivo an increase in glucose transport with insulin in non-diabetic mice and a defect of glucose transport in type 2 diabetic mice. It is the first time that an iodinated analogue of glucose has shown such promising results after in vivo injection. The use of this tracer to assess glucose transport in vivo in humans via nuclear imaging warrants further investigation.

Initial assessment of positron emission tomography for detection of nonpalpable regional lymphatic metastases in melanoma.

BACKGROUND: The purpose of this pilot study is to determine the feasibility of positron emission tomography with fluorodeoxyglucose (FDG-PET) for detection of nonpalpable regional lymph node metastases in patients with melanoma. METHODS: Adult patients with histologically proven cutaneous melanoma planned to undergo surgical lymphadenectomy for treatment of nonpalpable subclinical or residual metastatic melanoma in regional lymph node basin(s) participated. Each patient underwent attenuation-corrected PET imaging of the regional lymph node basin(s) with F18 fluorodeoxyglucose (FDG) followed by complete surgical lymphadenectomy. FDG-PET scans were interpreted prospectively by an experienced nuclear medicine physician. FDG-PET scan interpretations and histologic results were then correlated. RESULTS: Eleven patients underwent 12 FDG-PET scans followed by 12 operations to clear 14 regional lymph node basins. FDG-PET correctly predicted the presence of metastatic melanoma in seven of seven surgical specimens. FDG-PET scans correctly predicted the absence of disease in seven of seven histologically negative node basins. Sensitivity was 1.0; specificity was 1.0. CONCLUSIONS: This study suggests that increased fluorodeoxyglucose uptake in palpably unremarkable regional lymph node basins in patients with melanoma is highly suggestive of metastatic disease.

Assessment of cancer recurrence in residual tumors after fractionated radiotherapy: a comparison of fluorodeoxyglucose, L-methionine and thymidine.

UNLABELLED: This study evaluates the midterm follow-up of tumor and normal tissue uptake of deoxyglucose, thymidine and methionine after fractionated radiotherapy to assess cancer recurrence in residual tumors. METHODS: AH109A tumor-burdened rats were treated with one to eight doses of 5Gy 60Co radiation. Tissue distribution study with 18F-FDG, 3H-thymidine and 14C-methionine, double-tracer autoradiography with 18F-FDG and 14C-methionine, and single-tracer autoradiography with 14C-labeled deoxyglucose, thymidine and methionine were performed 6 days after the end of therapy. RESULTS: Dose response study shows a significant decrease of tumor uptake of all tracers after two and more doses, even in the case of later recurrence. Whereas 3H-Thd and 14C-Met tumor uptake was similar to that of normal muscle, 18F-FDG tumor uptake remains higher than that of muscle, even in the case of complete tumor cure. The irradiated muscle shows a higher 18F-FDG uptake than the nonirradiated muscle. Autoradiography after eight doses (100% tumor cure) reveals elevated 14C-DG tumor uptake to be ascribable to nonmalignant cellular elements, in particular to a macrophage layer at the rim of necrotic areas. Autoradiography after four and six doses (33% and 57% tumor cure) shows the highest methionine and thymidine uptake in viable cancer cells, whereas deoxyglucose uptake did not differ between viable cancer cells and macrophages. CONCLUSION: To detect and differentiate viable cancer cells in a residual tumor mass after radiotherapy, PET using 11C-methionine or 11C-thymidine may have some advantages over 18F-FDG, especially if the residual tumor includes larger areas of necrosis.

Differentiating benign from malignant lung lesions using "quantitative" parameters of FDG PET images.

UNLABELLED: Fluorine-18 labeled deoxyglucose positron-emission tomography (FDG-PET) applications in oncology include the differential diagnosis of chest masses and single pulmonary nodules. However, FDG is not tumor-specific; rather, it also accumulates in inflammatory processes. This study was performed to identify image parameters that would improve the specificity of PET. METHODS: Twenty-six patients who had benign and malignant lung lesions were examined retrospectively. Positron-emission tomography data were acquired in dynamic scanning mode after intravenous bolus of 250-402 MBq of FDG. Standardized uptake values (SUVs) were calculated and Patlak analyses were performed in selected regions of interest in the PET images. Positron-emission tomography results were related to histological diagnosis (N = 49) or clinical follow-up (N = 3). RESULTS: The specificity and sensitivity of the original PET scan reports, which was based on visual image interpretation and loosely applied SUVs, was 100% and 73%, respectively. Using the SUVs with a cut-off value of 3.8 and Kpat value with a cut-off at 0.025 min-1 improved the specificity to 81% and 85%. CONCLUSION: FDG-PET image interpretation can be facilitated by using SUV information or the accumulation rate of the radiotracer (Patlak). With additional validation, this method could have a significant cost-effective impact on the medical/surgical management of chest masses.

A modeling method to improve quantitation of fluorodeoxyglucose uptake in heterogeneous tumor tissue.

UNLABELLED: The standard approach for evaluating FDG-PET kinetic studies is based upon an assumption that tissue within a representative region of interest (ROI) is relatively homogeneous in terms of FDG kinetics. In neoplasms and other disease states, tissue within an ROI may be grossly heterogeneous, due to adjacent infarcted tissue and other causes. We have developed a method employing two ROIs (one over the tumor and another over a "reference region") to deal with this level of heterogeneity. METHODS: The method is based on the regular FDG model but consists of six variable parameters (6P model) which uses the kinetics in the reference region to account for the normal tissue within the tumor ROI, so that the kinetic data only associated with the tumor can be estimated. Monte Carlo simulations and human PET FDG studies were used to analyze the performance of the 6P model. RESULTS: The narrower 95% confidence intervals of parameter estimates, which centered at the true tumor rate constants, and the smaller correlation matrix of the 6P model showed the better performance of the 6P model compared to the standard "homogeneous" four-parameter FDG model. Computer simulations further showed that the 6P model can accurately estimate the microparameters (rate constants: K1* (ml/min/g), k2* (min-1), k3* (min-1), k4* (min-1)) and the macroparameter (K (ml/min/g)) of tumor cells regardless of the percent weight of tumor cells in the lesions. CONCLUSIONS: The new method can produce more reliable and accurate estimates of tumor glucose metabolic rates with dynamic PET FDG studies.

Brain activity patterns in flying, echolocating bats (Pteronotus parnellii): assessment by high resolution autoradiographic imaging with [3H]2-deoxyglucose.

Brain activity patterns during echolocation and flight were assessed in mustached bats (Pteronotus parnellii parnellii). Bats were injected intraperitoneally with [3H]2-deoxyglucose and restrained in a foam holder or allowed to fly for 20 min. Under resting conditions, low levels of [3H]2-deoxyglucose uptake were observed throughout the forebrain but relatively high uptake was found in brainstem auditory and vestibular centers. In flying, echolocating bats, marked increases in regional [3H]2-deoxyglucose uptake were apparent. All structures of the classical ascending auditory pathway were intensely labeled in autoradiograms. Other brain regions that exhibited high [3H]2-deoxyglucose uptake in flying bats included the cingulate cortex, stratum lacunosum-moleculare of the hippocampus, thalamus, caudate-putamen, superior colliculus, pontine reticular formation, nucleus ambiguus, parts of the midbrain central gray, and cerebellum. In the cerebellum, the most prominent increase in [3H]2-deoxyglucose uptake was found in discrete patches of the granule cell layer. The results provide the first overview of brain activity patterns during echolocation and flight in bats. In addition, uptake of [14C]fluorodeoxyglucose was used to compare brain activity patterns in flying bats to bats that were imaging their environment via biosonar while hanging in a wire cage. The echolocating-not-flying bats emitted 6931 +/- 1226 pulses in 20 min compared to 8972 +/- 1273 pulses in 20 min for flying bats. The uptake of the metabolic marker was significantly more in the flying bats compared to the emitting-not-flying bats in the medial geniculate, superior colliculus, auditory cortex, cingulate cortex and thalamus. In the nucleus ambiguus, cochlear nucleus, and inferior colliculus, uptake was similar for the flying and emitting-not-flying bats. These results suggest that the high metabolic activity observed in forebrain auditory regions of flying bats is related in part to neural processes that involve sensory motor integration during flight and not simply the perception of acoustic information.

In vitro assessment of 2-fluoro-2-deoxy-D-glucose, L-methionine and thymidine as agents to monitor the early response of a human adenocarcinoma cell line to radiotherapy.

The tumor cell uptake of three tracers that can be labeled with isotopes suitable for PET imaging--FDG, L-methionine and thymidine--were examined in vitro in a human ovarian carcinoma cell line (HTB77IP3) at varying times following 30 Gy 60Co irradiation and were compared to a nonirradiated control group. FDG, methionine and thymidine uptake per tissue culture well all increased following irradiation when compared to basal values, although to a much lower extent than the increases in uptake seen in a nonirradiated group. This increase in tracer uptake occurred despite a 6.25-fold decline in viable cell numbers. When examined per cell, FDG uptake per cell increased 9.77-fold, methionine 7.82-fold and thymidine 9.48-fold over basal levels from Day 0 to Day 12 following irradiation. Part of these increases may be due to giant cell formation and/or radiation repair processes that require energy, protein and DNA substrates. While the in vitro system differs from in vivo systems due to the absence of a blood supply in vitro, a lack of infiltrating leukocytes and other factors, our data suggest that early assessment of human adenocarcinoma response to radiotherapy by PET with these tracers may be complicated by this normal increase in tracer uptake postirradiation. Clearly, in this human cancer cell line, early radiation-induced cell death is not associated with an early decline in tumor cell uptake of FDG, methionine or thymidine.

Assessment of the role of adrenoceptor function in ischemia-induced impairment of 2-deoxyglucose uptake and CA1 field potential in rat hippocampal slices.

The release of catecholamines, dopamine and noradrenaline has been suggested to play a role in mediating ischemic damage in susceptible brain regions, the hippocampus and striatum. We now provide evidence that suggests a role for adrenoceptors in the deficit of 2-deoxyglucose uptake and CA1 field potential induced in hippocampal slices by hypoxia/hypoglycemia (ischemia). Treatment with alpha 1- or beta-adrenoceptor agonists or cAMP potentiated an ischemia-induced decline of both 2-deoxyglucose uptake and CA1 field potential in hippocampal slices, whereas alpha 1- or beta-adrenoceptor antagonists, or alpha 2-adrenoceptor agonists produced a remarkable neuroprotective action against these deficits. The results indicate that stimulation of adrenoceptors may play a detrimental role in the development of ischemic damage, and suggest a neuroprotective action for adrenoceptor antagonists, which may lessen the functional deficits induced by ischemia.

Evaluation of liver tumors using fluorine-18-fluorodeoxyglucose PET: characterization of tumor and assessment of effect of treatment.

To evaluate glucose metabolism in patients with tumors involving the liver, 35 patients with liver lesions had PET using 18F-2-fluoro-2-deoxy-D-glucose (FDG). FDG (148 MBq) was injected and radioactivity of the tumor was scanned dynamically by PET. The rate constants (k1, k2, k3, k4) of FDG in a metabolic model were calculated. The results were compared to hexokinase activity in the excised tumor specimens. k3 was found to reflect tumor hexokinase activity. When k3 was used as an index (cut-off value: 0.025), it was possible to distinguish benign and malignant tumors. k4 was significantly higher in hepatocellular carcinoma. By using k3 and k4 as indices, one could assess the degree of differentiation of hepatocellular carcinoma. After treatment, k3 decreased according to the effectiveness of therapy and thus may be a useful index for quantitatively assessing tumor viability.

Assessment of accuracy of PET utilizing a 3-D phantom to simulate the activity distribution of [18F]fluorodeoxyglucose uptake in the human brain.

A three-dimensional brain phantom has been developed to simulate the activity distributions found in human brain studies currently employed in positron emission tomography (PET). The phantom has a single contiguous chamber and utilizes thin layers of lucite to provide apparent relative concentrations of 5, 1, and 0 for gray matter, white matter, and CSF structures, respectively. The phantom and an ideal image set were created from the same set of data. Thus, the user has a basis for comparing measured images with an ideal set that allows a quantitative evaluation of errors in PET studies with an activity distribution similar to that found in patients. The phantom was employed in a study of the effect of deadtime and scatter on accuracy in quantitation on a current PET system. Deadtime correction factors were found to be significant (1.1-2.5) at count rates found in clinical studies. Deadtime correction techniques were found to be accurate to within 5%. Scatter in emission and attenuation correction data consistently caused 5-15% errors in quantitation, whereas correction for scatter in both types of data reduced errors in accuracy to less than 5%.

Clinical assessment of therapeutic effects on cancer using 18F-2-fluoro-2-deoxy-D-glucose and positron emission tomography: preliminary study of lung cancer.

Using positron emission tomography, we studied the tumor uptake of 18F-2-fluoro-2-deoxy-D-glucose (18FDG) in five lung cancer patients before and after anti-cancer therapy (radiotherapy and/or chemotherapy). The tumor uptake of 18FDG was classified as positive and negative; the former, by increasing the uptake of 18FDG with time, and the latter, by decreasing or the constant uptake of 18FDG. Before therapy, all cases tested positive. After therapy, three cases were negative and two cases remained positive. All negative cases corresponded to complete second 18FDG study. Our findings in the 18FDG study correlate with the clinical results. 18FDG is a promising method for assessing therapeutic effects on cancer clinically.

Assessment of hepatic indicators of subchronic carbon tetrachloride injury and recovery in rats.

To determine the course of hepatic recovery from subchronic oral administration of carbon tetrachloride (CCl4), male F-344 rats were gavaged with 0, 20, or 40 mg CCl4/kg, 5 days/week, for 12 weeks. Exposure to CCl4 caused dosage-dependent increases in relative liver weight and the serum levels of aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, alkaline phosphatase, and cholesterol as well as a dosage-dependent decrease in hepatic cytochrome P450. Centrilobular hepatocellular vacuolar degeneration, necrosis, and cirrhosis occurred at both 20 and 40 mg/kg, with dosage-dependent severity. Reversibility of these reported effects varied with parameter. By Day 8 postexposure, necrosis had disappeared and all serum indicators and cytochrome P450 had returned to control levels. By Day 15 postexposure, the severity of the vacuolar degeneration had decreased. Reversibility of cirrhosis was dosage dependent; complete recovery occurred in the low- but not the high-dose group by Day 15. The disappearance of the increase in relative liver weight was also dependent on dosage; the low- but not the high-dose group had returned to the control level by Day 22. In an attempt to measure persistent hepatic damage, liver uptake relative to the spleen was determined for a sulfur colloid labeled with technetium-99m and for tritiated 2-deoxyglucose. Neither method consistently measured hepatic damage in cirrhotic livers due, in part, to the high degree of variability in the tracer uptake data.

[PET assessment of myocardial viability with glucose loading and fasting FDG images].

Positron emission tomography (PET) offers the potential capability of evaluating tissue viability. We have studied the changes of myocardial F-18 deoxyglucose (FDG) uptake with glucose loading. In a fasting state (for at least 5 hours) and in a glucose loading state (50 g glucose orally one hour before the study) FDG (74-148 MBq) PET studies were performed for 50-60 minutes in 3 cases. 2 were subjects with anterior myocardial infarction (MI) including one with a ventricular aneurysm (case 1), one with a recent MI (case 2). One was a subject of aortic valvular disease without coronary lesions (case 3). Arterial input function (Ca(t)) and myocardial activity (Cm(t)) were derived from the regions of interest (ROI) on the left atrium and from multiple ROI's circumferentially about the myocardium. Net extraction, FU (Fractional Uptake) = Cm(T)/integral of T0 Ca(t)dt, were calculated. Normal segment (N) showed an increase in FU with glucose loading, but the ischemic segment showed no increase (case 1) or a lower increase (case 2) relative to N. This study shows that the increase in FU with glucose loading suggests the persistence of viable myocardium. In conclusion, myocardial viability may be evaluated by comparing FDG images in a glucose loading state with those in a fasting state.

Topographical assessment of accumulated radioactivity from [14C]2-deoxyglucose and [6(-14C)]glucose in rat forebrain at different survival periods.

The uptake and retention of radioactivity was measured in discrete areas of rat brain at different times after i.v. injection of [14C]2-deoxyglucose or [6(-14)C]glucose, in unrestrained rats. In most brain regions, the accumulation of radioactivity from the two compounds was similar when a 30-min survival period for [6(-14)C]glucose was compared to a 45-min survival period for [14C]2-deoxyglucose. However, at those times, autoradiographic images of the hippocampus and piriform cortex appeared distinctly different for [14C]2-deoxyglucose and [6(-14)C]glucose. Relatively more radioactivity accumulated from [14C]2-deoxyglucose, compared to [14C]glucose, in the stratum lacunosum-moleculare of the hippocampus and in layer 4 of the isocortex. In contrast, relatively more radioactivity accumulated from [6(-14)C]glucose, compared to [14C]2-deoxyglucose, in the molecular and granule cell layers of the dentate gyrus, the CA1 pyramidal cell layer of the hippocampus, and in layer 2 of the piriform cortex. When rats were killed 5 min after injection of [6(-14)C]glucose, the relative neuroanatomical distribution of radioactivity was similar to the 30-min survival period, except in layer 4 of the isocortex, where relatively more radioactivity was present at the early time. When rats were killed 5 min after injection of [14C]2-deoxyglucose, in 20 of 24 brain regions examined, the absolute and relative amounts of accumulated radioactivity were similar when compared to that of the 45-min survival period. In contrast, the absolute and relative amounts of radioactivity were significantly greater for the 5-min compared to the 45-min survival period, in the CA1 pyramidal cell field, dentate gyrus, and layer 2 of the piriform cortex. For those regions, the appearance of autoradiograms prepared from rats killed 5 min after administration of [14C]2-deoxyglucose is remarkably similar to the appearance of autoradiograms prepared from rats killed 5 or 30 min after injection of [6(-14)C]glucose. Possible mechanisms are discussed to explain the observed differences in the accumulation of radioactivity in discrete brain regions after injection of [6(-14)C]glucose and [14C]2-deoxyglucose at the different survival times examined.