Kinetics of intracolloidal iodine in thyroid of iodine-deficient or equilibrated newborn rats. Direct imaging using secondary ion mass spectrometry.

The most significant impact of the Chernobyl accident is the increased incidence of thyroid cancers among children. In order to accurately estimate the radiation dose provided by radioiodines, it is important to examine how the distribution of newly incorporated iodine varies with time and if this distribution varies according to the iodine status. The kinetic distribution of intra colloidal newly organified iodine in the rat immature thyroid was recorded and analysed using the ionic nanoprobe NanoSims50. Our observations imply that in case of radioiodine contamination, the energy deposits vary (i) with time, (ii) from one follicle to another, and (iii) from one cell to another inside the same follicle regardless the iodine status. The kinetic heterogeneity of iodine distribution must be take in account in thyroid dose evaluation.

Microautoradiographic study of technetium-99m colloid uptake by the rat liver.

A new microautoradiographic technique was developed to study the distribution of 99mTc-labeled radiopharmaceuticals. Using a thick emulsion, it is possible to get microscopically visible tracks of internal conversion and Auger electrons. The liver uptake of microscopic particles has been thought to occur in Kupffer cells but no direct evidence has been provided for technetium colloids. Using this method, 99mTc-labeled colloids were clearly identified in Kupffer cells in the sinusoidal areas of liver. "Track" microautoradiography using a thick emulsion layer may be used on any frozen tissue sections and may provide an important tool to assess the biodistribution of 99mTc radiopharmaceuticals.

Dosimetry at the cellular level of Kupffer cells after technetium-99m-sulphur colloid injection.

The radiation dose to Kupffer cells was estimated at the cellular level after intravenous injection of 99mTc labeled sulphur colloids in rats. The results were then compared with those obtained using macroscopic dosimetry. From the microscopy appearance observed using a "track" microautoradiographic method (MAR), it was shown that only 0.2% of the Kupffer cells were actually involved in the pinocytosis of radioactive colloids. For each electronic emission from 99mTc (Auger and internal conversion), the fraction of the emitted energy actually absorbed within the Kupffer cell was calculated using the values provided by Berger. About 15% of the total energy emitted by electrons was absorbed in 0.2% of the Kupffer cells. If these results are extrapolated to humans, the dose absorbed by the labeled cells can be estimated to be between 0.5 and 0.9 Gy/MBq. This represents about 15,000 times the average electron dose to the liver as estimated from macrodosimetric methods. In cases such as this one where an important distribution heterogeneity is expected, dosimetric estimations at a cellular level may be particularly useful.

Heterogeneous distribution of technetium-99m-labeled microspheres in rat lungs: microautoradiographic evidence and dosimetric consequences.

UNLABELLED: The heterogeneity of 99mTc-labeled microspheres distribution within rat lung was visualized and quantified using a microautoradiographic "track" method (MAR). METHODS: MAR was used to study the uptake of radioactivity by individual microspheres, thereby enabling calculation of the range of particle activity. MAR was also used to visualize in rat lung sections the intrapulmonary distribution of the microspheres within the lungs after intravenous administration. The mean doses delivered to the cells in close contact with the labeled microspheres were calculated taking only the 99mTc electron emissions into account. RESULTS: All the microspheres were labeled. Nevertheless, the spectrum of visible tracks varied by a factor of 10, inducing a variable activity per microsphere from < 36 Bq to 325 Bq (mean activity-94 Bq/microsphere). No correlation existed between the radioactivity uptake and the size of microspheres. A very heterogeneous tridimensional distribution of the microspheres within the lungs were demonstrated with interparticle distances ranging from 57-4400 microns. On the other hand, only 1 of 2000 rat lung capillaries was obstructed. Using the mean activity, calculated delivered doses were found to reach approximately 6 Gy for the closest endothelial cells and 2 Gy for epithelial cells. However, such high doses were delivered to only a few cells. CONCLUSION: The number of obstructed capillaries in human lungs is lower than in rat lungs; the distances between microspheres should be larger. Nevertheless, the individual doses absorbed by the pulmonary cells closest to the microspheres should be very important.

Active intestinal elimination of ciprofloxacin in rats: modulation by different substrates.

1. Two in vivo models, in the rat, were used to investigate, in the presence of different substrates, the overall and net intestinal elimination of ciprofloxacin: an open-intestinal perfusion model and an intestinal loop model respectively. 2. In the presence of quinidine, verapamil and cyclosporin (substrates of the P-glycoprotein (P-gp)), plasma AUCs of ciprofloxacin were 1.5 - 2 fold increased, while biliary clearance (1.5 - 2 fold), intestinal overall and net clearances (2 - 4 fold and 1.5 - 8 fold respectively) decreased. The weak effect obtained with cyclosporin as compared to verapamil and especially quinidine, suggests, for ciprofloxacin, the existence of transport systems distinct from the P-gp, as the OCT1 transporter which could be inhibited by quinidine. 3. With cephalexin and azlocillin, two beta-lactam antibiotics, plasma AUCs of ciprofloxacin increased and biliary and intestinal overall clearances decreased in a similar fashion (1.3 - 2 fold), suggesting the involvement of organic anion and/or cation transporters. 4. In the presence of structural analogues, the effect was dependent on the compound administered: Sparfloxacin had no effect on intestinal clearance of ciprofloxacin. In contrast, with pefloxacin, overall intestinal clearance of ciprofloxacin was decreased and net intestinal clearance increased. 5. The specificity of ciprofloxacin intestinal transport appears to be different from P-gp as outlined by the lack of competition with sparfloxacin, a P-gp substrate. Ciprofloxacin intestinal elimination seems to be mediated by organic anion and/or cation transporters and a mechanism sensitive to quinidine and verapamil.

Immunoreactive beta-endorphin increases after an aspartame chocolate drink in healthy human subjects.

It has been claimed that sucrose intake induces a rise in beta-endorphins. In an attempt to discriminate between the sensorial and metabolic effects of sucrose intake in this process, the effects of two chocolate drinks were compared: one sweetened with 50 g of sucrose, the other with 80 mg of aspartame. Plasma beta-endorphin concentrations were more elevated after the aspartame drink than after sucrose or fasting, while insulin increased after drinking as much with aspartame as with sucrose. We suggest that the increase in beta-endorphin after aspartame edulcorated chocolate is related with insulin secretion in the absence of marked changes in blood glucose or with a direct effect of aspartame itself on beta-endorphin liberation.

Lack of plasmic beta-endorphin response to a gastronomic meal in healthy humans.

In order to study the relationship between the endogenous opiate system and food intake in man, plasma concentrations of beta-endorphin were measured in ten healthy subjects. Time course of beta-endorphinemia was compared under the following conditions: basal (fasting), after an injection of pentagastrin (6 micrograms/kg), or after a gastronomic meal. No changes in plasma beta-endorphin or ACTH concentrations were observed with pentagastrin nor after the meal, despite the combination of very high sensory pleasure with intake of a very large amount of food. It is concluded that blood beta-endorphin concentration is not a sensitive index of the effects of food intake on the endogenous opioid system in man.

Technetium 99m labelled heparin: pharmacokinetics and tissue distribution in rats after vascular surgery.

Pharmacokinetics of technetium 99m (99mTc) labelled heparin was studied after I.V. infusion (70 IU/kg) in normal rats and in a group of animals just after aortic longitudinal incision and suture. Anticoagulant activity measurements and radioactivity were compared on the same plasma samples. The pharmacokinetic decay of 99mTc-heparin followed a bi-exponential pattern. Half-lives (T1, T2) were significantly shortened after surgery using both techniques. Liver accumulation of labelled heparin was decreased in "sutured" rats and a high uptake of 99mTc-heparin was found on the operated area of the aorta.

Pharmacokinetics and tissue distribution of 99mTc-labelled enoxaparin in the rat: evaluation of dosimetry parameters.

A low molecular weight heparin, enoxaparin, was labelled with 99mTc and the characteristics of the labelled compound determined. In vitro the stability, and labelling efficiency (98%) of the labelled drug were excellent. Rats were injected with 99mTc-enoxaparin to study pharmacokinetics and distribution. The results were used to calculate dosimetric estimates which are a prerequisite for pharmacokinetic studies on labelled LMWH (low molecular weight heparin) in human subjects. Biodistribution studies showed preferential liver and spleen accumulation. But the doses absorbed by these target organs remained below the upper limits of the dose received by a patient undergoing hepatic scintigraphy.

Quantitative autoradiography using a radioimager based on a multiwire proportional chamber.

Determination of the biodistribution of radiopharmaceuticals is an important issue for the evaluation of their performance in diagnosis and therapy. In this study, we evaluated a digital radioimager (RI) based on a multiwire proportional chamber for quantitative autoradiography (AR). The RI allows direct detection of electronic emissions of gamma emitters. Its qualitative and quantitative performances were tested on 99mTc and (111)In labelled sections and compared with conventional film AR. Linearity of count rate versus activity was verified over a 104 range of activity. As compared with film AR, a substantial improvement of the detection limit was obtained even for acquisition periods up to 20 times less than film exposure times. We provided the basis for quantitative analysis with tissue equivalent paste standards: the 99mTc and (111)In RI counting efficiencies were respectively 1.19% and 2.35%. We illustrated the respective values of RI and film AR in two rat studies: 99mTc-DMSA in kidney and dual-isotope 99mTc-MIBI and (111)In-antimyosin in heart. Calculated activity concentrations on sections of rat organs confirmed good correlation to gamma counting (deviation less than 12%). We suggest RI as a convenient technique for fast localization of single or dual-isotope tracers and determination of activity distribution.

[Experimental arterial microsutures. Evaluation of the risks of thrombosis using technetium 99-labeled heparin]. / Microsutures artérielles expérimentales. Evaluation des risques de thrombose par l'héparine marquée au technétium 99.

Following demonstration of a method of assay of circulation heparin by measurement of radioactivity after labelling with 99 m technetium, the technique was used to study effects of arterial microsuture on metabolism of heparin injected postoperatively. Results of a study involving rat aorta showed that local binding was maximum during the first half-hour and that abnormal consumption of heparin continued during the 6 hours following operation. These finding suggest that a risk of thrombosis probably persists throughout this postoperative period.

Thallium uptake in fasted and nonfasted rats.

Thallium uptake was studied in selected organs in two groups of rats: fasted and nonfasted. A significant increase in thallium uptake was found in the stomach and the small intestine after eating while a significant decrease was found in the lung.

Mutual contribution by "track" microradioautography method (MRA) and secondary ion mass spectrometry (SIMS) microscopy: prospects in technetium dosimetry at-the cellular level.

The determination of cellular uptake sites of radioligands used for cell labelling for diagnostic purposes is an essential prerequisite for evaluating the radiation dose to the cell nucleus and cytoplasm. The distribution of 99mTc-HMPAO in labelled leukocytes was studied by two microscopic imaging techniques on the same biological material: the "track" microradioautographic method (MRA) and Secondary Ion Mass Spectrometry (SIMS) microscopy. The "track" method used internal conversion electrons of 99mTc, leading to the formation of silver grains in a thick layer nuclear emulsion deposited onto cellular smear. In order to improve the specificity of the "track" detection, a minimum of 5 consecutive silver grains was required. In SIMS Microscopy, mapping with 99Tc "daughter" nuclide of 99mTc (half-life: 2.13.10(5) years) was obtained after sputtering of superficial molecular layers on embedded specimen sections. A mass resolution of about 5,000 was needed to circumvent polyatomic ion interferences. Both methods were able to demonstrate a very heterogeneous distribution of technetium from one cell to another. The sensitivity and signal/noise ratios were excellent for both methods. The lateral resolution of SIMS microscopy (0.5 microns) was far better than that of MRA. Therefore, only SIMS is able to distinguish between nuclear and cytoplasmic localization. On the other hand, quantification was not achieved for SIMS, although semi-quantification is possible with MRA. The field of view of MRA is far larger, allowing a better statistical approach for quantification. Both methods appear to be complementary to determine the distribution of technetium at the cell level. MRA is simpler and better fitted to the study of a cell population or a tissue. The unique spatial resolution of SIMS allows to focus the study on subcellular structures.

[Microautoradiographic method permitting the study of migration of 99m-technetium labelled leukocytes]. / Sur une méthode de microautoradiographie permettant l'étude de la migration des leucocytes marqués par le technétium-99m.

Cell migration studies are very important in inflammatory phenomena. Methods currently used are not quantitative and have been subject to much controversy. Homogeneity of 99mTc leukocyte labelling was verified by a microautoradiographic method (MAR), which was performed in our laboratory. This method was used in cell migration studies to verify if the migrating cells were indeed the labelled cells.

Biological consequences of irradiation by low doses of technetium 99m: ultrastructural studies, p53 protein expression and cytogenetic effects.

Few studies concerning the potential genetic effects of diagnostic radionuclides used in nuclear medicine have been reported. The aim of this study was to evaluate the biological and cytogenetic consequences of two technetium 99m-labelled radiopharmaceuticals. Ultrastructural modifications of pulmonary cells were first investigated after injection of 99mTc labelled microspheres in the rat. On the same irradiated cells, nuclear expression of p53 protein was assessed using immunohistochemistry. Despite very high previously calculated doses delivered to pulmonary cells, no morpholological cell damage and no significant increase of nuclear expression of the p53 were noted. There was no correlation between the calculated dose and the ultrastructural biological damage. Secondly, a specific in vitro curve, activity/number of unstable chromosomal aberrations, corresponding to physical characteristics of 99mTc, was established to verify the potentiality of 99mTc to induce such aberrations. In vivo, cytogenetic effects were assessed on blood samples of 5 patients with various arthrosic and periarthrosic diseases obtained after bone scintigraphy. Aberration frequencies of both in vitro and in vivo irradiated lymphocytes were determined using the classical Fluorescence Plus Giemsa technique. No cytogenetic effects appeared with the routinely 99mTc injected activities as predicted by the in vitro curve.

The perinatal thyroid in iodine deficient regions: risks of radioiodines--hazards of stable iodine overload. A study in the newborn rat.

Administration of large quantities of stable iodine is an effective means of reducing the radiation burden on the thyroid in the event of a nuclear power-plant accident. Such administration may involve countries with low baseline dietary iodine intake. It is questioned whether stable iodine overload is safe, and in particular, what are its effects in newborn infants? Iodine-deficient newborn rats were submitted to a single acute administration of stable iodine (100 microg) on the second day of life. The effects on thyroid structure were studied, after 24 hr and after 7 days, using light microscopy. Compared to controls, the thyroids of animals submitted to stable iodine overload showed, 7 days after treatment, signs of acute toxicity including marked desquamation of epithelial cells and rupture of a large number of thyroid follicles. Our findings in iodine deficient newborn rats suggest that stable iodine overload may have side effects during perinatal life. This prophylactic measure should, therefore, be accompanied by follow-up of thyroid function. Thyroid hormones are critical for brain development, during the first period of life.

Tissular distributions and kinetics of two renal 99mTc-radiopharmaceuticals in rat.

A digital radioimager (RI), conventional radioautography (RA), and tracks microradioautography (MRA) were used to assess the biodistributions and kinetics of 99mTc-dimercaptosuccinic (99mTc-DMSA) and 99mTc-mercaptoacetyltriglycine (99mTc-MAG3) in rat at both macroscopic and microscopic levels. Three groups of male Wistar rats were studied. Using gamma-counting, kidney, liver, spleen and blood kinetics of both tracers were assessed in the three groups. Using RA and RI, renal slices were analyzed in group 1 the animals being sacrified from 2 to 60 min after injection of 99mTc-MAG3, and in group 2 the animals being sacrificed from 0.5 to 24 hr after injection of 99mTc-DMSA. Using MRA, renal slices were analyzed for 99mTc-DMSA (group 3). RA films and RI images displayed the variation with time of the cortical and medullary uptakes of the tracers. No regional heterogeneity within the different structures could be seen neither with RA films nor with MRA. The remaining activity in the blood 24 hr after injection of 99mTc-DMSA was evaluated. The tissular distributions of both tracer being homogenous, mean values of cortical uptake seems to be acceptable for dosimetric studies. Our results incite to use of 99mTc-MAG3 instead of 99mTc-DMSA when both tracers may be indicated.

In vitro irradiation of blood with 99mTc: evaluation of dose and chromosome aberrations in irradiated lymphocytes.

The use of ionizing radiation for diagnostic or therapeutic purposes in medicine represents the principal source of artificial radiation to humans. Calculation of radiation dose is essential to the analysis of risks (biological effects) and benefits in any application, including nuclear medicine. The dose assessment in many cases is not necessarily straightforward. Many radiopharmaceuticals are labelled with radionuclides that undergo not only gamma-emission but also emission of Auger and internal conversion electrons. A typical example is technetium-99m (99mTc), which is used in more than 80% of nuclear medicine applications. In this work, in vitro studies have been carried out to evaluate the dose delivered to lymphocytes by human serum albumin microspheres (HSAM) labelled with 99mTc. Experiments were performed in order to score unstable chromosomal aberrations induced by 99mTc-HSAM, using conventional cytogenetic techniques. Henceforth, the relationship between activities introduced into blood samples and induced chromosomal aberrations were evaluated. To assess the dose absorbed in lymphocytes, electron and photon transport was performed in a simple model representing the system used for irradiating the cells using the MCNP Monte Carlo code. In this report, analysis of dose-effect curve demonstrates a linear quadratic response for unstable chromosome aberrations.