Therapeutic radiopharmaceuticals.

A variety of radiopharmaceuticals have been introduced for the internal therapy of malignant and inflammatory lesions in nuclear medicine. In order to destroy the diseased tissues radionuclides with high linear energy transfer (LET) such as beta, alpha, Auger or low energy conversion electron emitters are needed. The range of beta particles is in mm's so they are effective for large tumors. The range of alpha particles is short, only a few cell diameters. Thus, they are effective in treating circulating malignant cells and micrometastases. The range of Auger and conversion electrons is <0.1 micro m. They are very effective in cell killing when they are carried across cell membrane into the nucleus to damage DNA. Appropriate ions, molecules and particles are labelled with such radionuclides and used as radiopharmaceuticals in many different applications. For an effective dose to be delivered high target to non target ratios must be attained. Monoclonal antibodies to specific antigens expressed on tumor cells have been developed to increase the uptake by malignant tissues by specific accumulation. Radiolabelled peptides such as somatostatin, small molecules such as metaiodo-benzylguanidine (MIBG) and many different nano-and micro-particles have been investigated. The effectiveness of therapy can be increased by direct locoregional administration of the radiopharmaceutical. This way the radiation effects are confined locally and the normal tissues are spared from radiation effects. In this review article selection criteria and characteristics of radionuclides and carrier ions, molecules and particles for various therapeutic applications will be discussed, including mainly the recent developments.

Radiopharmaceuticals for the visualization of infectious and inflammatory lesions.

Many radiopharmaceuticals have been introduced for the scintigraphic demonstration of infectious and inflammatory lesions and some of them are currently in clinical use. They can be classified into two major categories according to their specificity. Specific radiopharmaceuticals include in vitro labeled leukocytes, radio-labelled monoclonal antibodies, and receptor specific small proteins and peptides. Nonspecific radiopharmaceuticals include radiolabelled nanocolloids, liposomes, macromolecules such as human immunoglobulin, dextran and human serum albumin, various small molecules and ions. In this review article radiopharmaceuticals in their respective groups are discussed as to the efficacy, and other parameters such as the physical characteristics of the radionuclides used, radiochemistry involved, availability, cost and biodistribution, emphasizing recent developments.

Accumulation of some small molecular weight complexes of 99mTc in experimental abscesses.

The purpose of the present investigation was to evaluate the feasibility of imaging inflammatory lesions with some small molecular weight complexes of 99mTc with renal excretion in comparison to 99mTc-dextran (D), 99mTc-HIG and 67Ga-citrate. The biodistributions of pertechnetate (P), 99mTc-citrate (Cit), -gluconate (G), -glucoheptonate (GH), -DTPA, -glucose phosphate (GP), -D, -HIG and 67Ga-Cit were determined in mice with turpentine-induced abscesses at 1, 3, 6 and 24 h after i.v. injection of 15 MBq. % uptake/g tissues and abscess/muscle (A/M), blood (A/B), liver (A/L), intestine (A/J) and kidney (A/K) concentration ratios were calculated. The scintigraphic images of all mice were obtained by a 7-camera. The abscesses were well visualized with all the radiopharmaceuticals (RPs). The excretion was mainly via the kidneys except for 99mTc-D, 99mTc-HIG and 67Ga-Cit. They showed variable amounts of liver and kidney uptake. Similar A/M ratios were obtained with all the RPs, reaching max. at 3 h with pertechnetate, 99mTc-Cit and 99mTc-GP, at 6 h with 99mTc-DTPA and at 24 h with 99mTc-G, 99mTc-GH, 99mTc-D and 99mTc-HIG. The max. A/M ratios were 3.61 +/- 1.63, 4.61 +/- 3.92, 5.21 +/- 1.24, 3.60 +/- 0.52, 3.43 +/- 0.92, 5.37 +/- 0.67, 5.98 +/- 1.17 and 4.76 +/- 2.03 for 99mTc-P, -Cit, -G, -GH, -DTPA, -D, -HIG and 67Ga-Cit, respectively. Our results indicated that small molecular weight complexes of 99mTc can be used in imaging inflammation as well as high molecular weight complexes such as 99mTc-D, 99mTc-HIG and 67Ga-Cit. However, the first group is preferred, because of rapid blood clearance and excretion predominantly via the kidneys, which is an advantage for the identification of abdominal abscesses. The main mechanism of accumulation of all the RPs might be a simple diffusion process through the injured capillaries. An additional mechanism of binding to proteins was considered in view of the increasing concentration ratios with 99mTc-G, -GH, -D and -HIG as time progressed.

Experimental induction of osteosarcoma by subperiosteal radioactive phosphorus injections in rats.

In search of a predictable experimental model for the study of therapeutic modalities for osteosarcoma, a group of rats had injections of phosphorus-32 (P-32) colloid subperiostally and were followed for the occurrence of malignant bone tumors until death or for 18 months. Of 50 rats, 26 died of unrelated causes and only 24 were included in the study. Two cases of osteosarcomas, one occurring in month 16 and the other at month 18 were induced; one case demonstrated multiple lung metastases. Clear evidence of the sarcoma inducing effect of P-32 has been obtained in this study. However, a high yield experimental osteosarcoma model was not produced.

99mTc-citrate versus 67Ga-citrate for the scintigraphic visualization of inflammatory lesions.

Citric acid was labeled with 99mTc with an efficiency of > 99%. The biodistribution of 99mTc-citrate was studied in mice with turpentine-induced abscesses in comparison to 67Ga-citrate. The max. abscess/muscle concentration ratios were 4.61 +/- 1.92 (3 h) for 99mTc-citrate and 4.76 +/- 2.04 (4 h) for 67Ga-citrate. Arthritis was induced in 10 rabbits by intra-articular injection of ovalbumin Scintigrams obtained 4 days later and at 3 h post-injection of 99mTc-citrate showed increased activity involving the synovium. The max. arthritic/contralateral knee ratio was 3.19 +/- 1.29 (3 h) and 6.47 +/- 3.71 (24 h) for 99mTc- and 67Ga-citrate, respectively. The blood clearance curve of 99mTc-citrate in rabbits was biexponential with a fast (T1/2 = 36 min) and a slow (T1/2 = 18 h) component, compared to mono-exponential clearance of 67Ga-citrate (T1/2 = 23 h). In 10 patients with rheumatoid arthritis whole-body scintigrams and spot images of involved joints indicated localization of the tracer in inflamed tissues. The mean target-to-soft tissue ratios were 3.04 +/- 0.81 and 4.95 +/- 2.56 for 99mTc-citrate and 99mTc-MDP, respectively. Renal clearance of radioactivity was evident from the scintigrams. Our results demonstrated that 99mTc-citrate is effective as a radiopharmaceutical for the visualization of inflammatory lesions and may be preferred to 67Ga-citrate due to the ideal physical characteristics of the radionuclide, easy preparation, low cost, early accumulation and the preference for the renal route of excretion.

Effect of wearing complete dentures on human palatal mucosal blood flow measured by 133Xe clearance.

In eight edentulous subjects, who had not previously worn dentures, blood flow was measured by 133Xe clearance after intramucosal injection. Upper and lower complete dentures were then constructed for these subjects, and 133Xe clearance was measured after the dentures had been worn for 7 and 40 days. Before the insertion of dentures the mean blood flow to palatal mucosa was 18.9 +/- 7.1 ml/100 g/min. Seven and 40 days after wearing dentures the mean flows to the palatal mucosa were 10.6 +/- 4.5 and 12.6 +/- 5.3 ml/100 g/min, respectively. The difference before and after wearing dentures was statistically significant (p less than 0.05). The subjects were asked not to wear the dentures for 24 h after the third visit on the 40th day whereupon the blood flow reached almost normal levels (16.2 +/- 5.45 ml/100 g/min). The findings show that wearing dentures hinders the blood supply to the palatal mucosa, and that after resting the oral tissues, blood flow may return to normal levels.

Diclofenac sodium incorporated PLGA (50:50) microspheres: formulation considerations and in vitro/in vivo evaluation.

Recently, considerable interest has been focused on the use of biodegradable polymers for specialized applications such as controlled release of drug formulations; meanwhile, microsphere drug-delivery systems using various kinds of biodegradable polymers have been studied extensively during the past two decades. Poly (lactide-co-glycolide) (PLGA) polymers have been proven to be excellent drug carriers for microparticulate systems due to their advantages, e.g. biocompatibility and regulatory approval. The administration of nonsteroidal anti-inflammatory drugs (NSAIDs) into the intra-articular cavity in patients with chronic inflammatory disease is complicated due to the short duration of effect. In the present study, controlled-release parenteral formulations of diclofenac sodium (DS), a commonly used NSAID, were prepared for intra-articular administration, and evaluated in vitro for particle size, yield, drug loading, surface morphology and release characteristics. For in vivo studies, Technetium-99m labelled polyclonal human immunogammaglobulin (99m Tc-HIG) was used as the radiopharmaceutical to demonstrate arthritic lesions by gamma scintigraphy. Evaluation of arthritic lesions post-therapy in rabbits showed no significant difference in the group treated with PLGA (50:50) (mw 34000) DS microspheres compared to control groups.

Evaluation of 99mTc-labelled vitamin K4 as an agent for testicular imaging.

Vitamin K4 was labelled with 99mTc with an efficiency higher than 97%. The compound was stable up to 24 h at room temperature, and its biodistribution in NMRI mice indicated its in vivo stability. Blood radioactivity levels were high over a wide range. 10% of the injected activity remained in blood after 24 h. Excretion was mostly via kidneys. Only the liver and kidneys concentrated appreciable amounts of radioactivity. Testis/soft tissue ratios were 1.4 and 1.57 at 6 and 24 h, respectively. Testis/blood ratios were lower than 1. In vitro studies with mouse blood indicated that 33.9 +/- 9.6% of the radioactivity was associated with RBCs; it was washed out almost completely with saline. Protein binding was 28.7 +/- 6.3% as determined by TCA precipitation. Blood clearance of 99mTc-K4 in normal subjects showed a slow decrease of radioactivity, reaching a plateau after 16 h at 20% of the injected activity. In scintigraphic images in men the testes could be well visualized. The right/left testis ratio was 1.08 +/- 0.13. Testis/soft tissue and testis/blood activity ratios were highest at 3 h. These ratios were higher than those obtained with pertechnetate at 20 min post injection. 99mTc-K4 appears to be a promising radiopharmaceutical for the scintigraphic visualization of testes.

Accumulation of 99mTc-glutathione in head and neck tumors.

Glutathione labelled with 99mTc was used to study blood clearance and normal distribution in 3 healthy volunteers and in 10 patients with biopsy-proven tumors in the head and neck region. Static scintigrams were obtained at 1, 3, 6, and 24 h. ROIs over tumors and normal soft tissues were compared to obtain T/N ratios. In normal subjects blood clearance reached a plateau at 6 h; no radio-activity accumulation in the head and neck region was observed. Only the cardiac blood pool, the liver, the kidneys and the urinary bladder were evident. Excretion was via the kidneys. Malignant tumors and metastases were well visualized in 7 patients (true-positive), starting at 1 h. The mean T/N ratio was 2.69 +/- 0.77. The best images were obtained at 3-6 h. 1 false-positive (granulamatous reaction), 1 false-negative (malignant epithelial tumor in the radix of tongue) and 1 true-negative (angiofibroma) results were obtained. 99mTc-GSH is a potential radiopharmaceutical for the scintigraphic visualization of head and neck tumors. Further clinical studies are warranted to show its sensitivity and accuracy.

Internal mammarial lymphoscintigraphy with SPECT after ultrasound-guided injection of 99mTc-dextran.

Twenty-two women with stage II or III breast cancer were evaluated by SPECT to determine the number, size, three-dimensional localization and depth from skin surface of the internal mammary lymph nodes for accurate radiotherapy portal planning. The results were also compared with those of planar imaging (PI). Two-step injections of 99mTc-dextran were made under ultrasound guidance into the anterior sheath of the M. rectus abdominis first at the ablation side. 1.5 h thereafter an anterior planar view was obtained to evaluate cross drainage (13.6% was observed). Then, the second injection was done at the opposite side in the same manner. Planar and tomographic images were obtained 1.5 h after the second injection. Similar values were obtained for the depth from skin surface, distance from the midline and diameter of the lymph nodes with both PI and SPECT. The total number of nodes in 22 patients detected by SPECT was higher (138) than that from PI (129), the difference being statistically significant (0.005 greater than p greater than 0.0005).

Accumulation of 99mTc-Cysteine in experimental inflammatory lesions in comparison to 99mTc-glutamine and 99mTc-HIG.

AIM: The aim of this experimental work was to investigate the efficacy of 99mTc-L-Cysteine (Cys) in scintigraphic visualization of inflammatory lesions in comparison to 99mTc-L-glutamine (G), and 99mTc-HIG. METHODS: In mice abscesses were induced by intramuscular injection of turpentine. Six days later mice were injected with 3.7 MBq of each agent and sacrificed in groups of three at 1, 3, 6 and 24 h. Scintigrams were obtained with a gamma camera. The organs, some blood, abscesses, some muscle and urine were removed, weighed and counted in a gamma counter. Percentage of uptake by organs and per gram tissues and abscess/normal tissue concentration ratios were calculated. Experimental arthritis was produced in 6 New Zealand rabbits by intraarticular injection of ovalbumin. Four days later 37 MBq of 99mTc-Cys and 99mTc-HIG were each i.v. administered to 3 rabbits. Scintigrams obtained at 1, 3, 6, and 24 h demonstrated the arthritic joints very well. ROI's over arthritic joints were compared to contralateral normal joints (A/C). RESULTS: In mice the abscesses were well visualized on all scintigrams. The maximum abscess/muscle ratios were 5.21 +/- 1.09 (6 h), 3.73 +/- 0.81 (3 h) and 5.98 +/- 1.17 (24 h) and the maximum abscess/blood ratios were 3.46 +/- 1.33 (24 h), 1.81 +/- 0.10 (6 h) and 0.914 +/- 0.351 (24 h) for 99mTc-Cys, 99mTc-G, and 99mTc-HIG, respectively. In rabbits the maximum A/C ratios were 2.61 +/- 0.53 (3 h) and 2.92 +/- 0.99 (24 h) for 99mTc-Cys and 99mTc-HIG, respectively. CONCLUSION: Our results indicate that 99mTc-Cys is a promising agent for imaging inflammatory lesions. It is preferred to 99mTc-HIG, because of higher concentration ratios attained earlier, lower blood background, lower cost and a simpler in-house preparation method.

Visualization of bone pathologies and lung cancer with 99mTc-glucose phosphate: a comparative study.

Glucose phosphate (GP) labelled with 99mTc was used to obtain scintigraphic images of bone lesions in one group of patients (n = 28) and of lung tumors in another (n = 35). All bone lesions detected by 99mTc-MDP were also demonstrated by 99mTc-GP; all lung tumors except 4 were detected by 99mTc-GP, the failure rate being about the same as that for 67Ga. The use of 99mTc-GP is preferable to that of 99mTc-MDP because the former does not accumulate in normal bone; the advantage of 99mTc-GP over 67Ga lies in its better physical characteristics and in the fact that the result of the study is available within a few hours rather than three days.

Evaluation of 99Tcm-labelled alpha-D-glucose 1-phosphate aerosols for SPECT ventilation lung imaging.

The purpose of the present investigation was to evaluate 99Tcm-labelled alpha-D-glucose 1-phosphate (GP) aerosols for single photon emission computed tomographic (SPECT) ventilation lung imaging in comparison to 99Tcm-diethylenetriaminepentaacetate (DTPA) aerosols. Ten normal nonsmoking male volunteers (aged 20-30 years) were included in this study after obtaining their informed consent. 99Tcm-GP, 30 mCi, in 2 ml was placed in the nebulizer (Venticis II) and inhalation continued for 5 min of normal breathing with oxygen flowing through. In 10 subjects dynamic images were obtained from the posterior position for 90 min with 45 frames on a 64 x 64 matrix by the use of a gamma camera. At the end of the dynamic study planar images of the lung (anterior, posterior and laterals) were recorded. Decay corrected clearance curves and kep values were obtained by the pulmonary epithelial programme and T1/2 values were calculated. The same procedure was followed by the use of 99Tcm-DTPA in the same subjects 2 weeks later. SPECT studies of the lung were performed in five subjects after inhalation of 99Tcm-GP aerosols. Clearance curves were monoexponential. The difference in T1/2 values between the right and left lungs was statistically insignificant (P > 0.10). The mean T1/2 values were 316.5 +/- 44.7 and 80.8 +/- 13.4 min for 99Tcm-GP and 99Tcm-DTPA, respectively. The difference was significant (P < 0.0005). On scintigraphic images 99Tcm-GP showed high alveolar deposition and low adhesion to major airways like 99Tcm-DTPA. However, it is preferred to 99Tcm-DTPA for SPECT studies because of its prolonged pulmonary clearance.

99Tcm-glutathione for imaging inflammatory lesions.

The purpose of this study was to demonstrate the accumulation of 99Tcm-glutathione (GSH) in experimental abscesses. The biodistribution of 99Tcm-GSH was determined in mice with turpentine-induced abscesses in comparison with 99Tcm-human immunoglobulin (HIG) at 1, 3, 6 and 24 h. Arthritis was induced by intra-articular injection of ovalbumin in the right front knees of 16 rabbits. Each radiopharmaceutical (RP) was administered intravenously to eight rabbits. Scintigrams were obtained and regions of interest (ROIs) over the involved and contralateral knees were compared. Blood clearance of both RPs was studied in 10 normal rabbits. Biodistribution studies indicated accumulation of 99Tcm-GSH in abscesses. Abscess to liver, muscle, intestine and blood ratios were higher at all time points compared to 99Tcm-HIG. The maximum abscess:muscle ratios were 7.57 +/- 0.73 and 5.98 +/- 1.17 at 24 h for 99Tcm-GSH and 99Tcm-HIG, respectively. Scintigraphic images demonstrated both the abscesses and arthritic joints very well. 99Tcm-GSH is superior to 99Tcm-HIG due to clearance predominantly via kidneys, no significant accumulation in any other organs, high target-to-nontarget ratios attained a few hours after administration, low cost and in-house preparation by a simple procedure.

Parasternal lymphoscintigraphy using 99Tcm-dextran.

99Tcm-dextran was evaluated as a lymphoscintigraphic agent in 10 normal volunteers and 24 patients with breast cancer. 99Tcm-dextran (0.5-1 mCi) in a volume of 0.1-0.2 ml was injected into the posterior rectus sheath in the subcostal site on one side. Scintigrams were obtained at 2-2.5 h after injection and the injection was repeated on the other side. The final image was taken at 4.5-5 h after the first injection. The normal distribution of lymph nodes and anatomical variations in scintigraphic images were first determined in normal subjects. In the patient group, eight scintigrams were evaluated as pathological and the rest as normal. In the 25 women whose lymphoscintigrams were normal the mean number of parasternal nodes visualized was six (range: 3-11). Bilateral symmetrical or asymmetrical chain was observed in 20 subjects (80%) and unilateral chain in five subjects (20%). There was cross-drainage in five subjects (20%). Supraclavicular lymph nodes were visualized in 11 subjects (44%). Varying abnormal images were obtained in eight patients. Our results are in good agreement with those reported in the literature, using radiocolloids. It is concluded that 99Tcm-dextran is a promising agent and may well replace radiocolloids because of the simplicity of the labelling procedure, stability of the label, easy availability and low cost.

Detection of head and neck cancer with 99Tc(m) glutathione: a correlative study with tissue glutathione and glutathione S-transferase levels.

In this study glutathione (GSH), a natural tripeptide which plays an important role in detoxification reactions, protecting cells against damage from xenobiotics, has been labelled with 99Tc(m) for the demonstration of head and neck cancer. Twenty-eight patients (10 females and 18 males) with various malignancies of the head and neck were given 740 MBq of 99Tc(m)-GSH intravenously and single-photon emission computed tomography (SPECT) images were obtained at 3 h. Semiquantification was performed by drawing regions of interest on three consecutive transaxial slices and tumour to background ratios were calculated. In addition, GSH and glutathione S-transferase (GST) levels were measured in the tumour samples and in normal tissue which were obtained during surgery. Scintigraphic images showed that there was increased uptake in the tumour compared to the normal contralateral side (tumour/normal tissue (mean +/- SD) = 1.94 +/- 0.76). The tissue analyses revealed increased levels of GST in tumour tissues, but both GST and GSH levels in tumour were not statistically different from those in the normal tissue. We conclude that scintigraphic visualization of head and neck tumours can be attributed to increased demand for GSH in cancer. Protein binding might account for the prolonged retention of 99Tc(m)-GSH in the malignant tissue. Like other peptides, it is accumulated and excreted by the kidneys, which allows clear visualization of the abdomen without interference from gastrointestinal system activity.

Evaluation of 99Tcm-HIG as a lymphoscintigraphic agent in rabbits.

In this pilot study, 99Tcm-labelled human immunoglobulin G (99Tcm-HIG) was evaluated as a lymphoscintigraphic agent in five rabbits. It was injected intradermally into the web space of the hind legs of the rabbits (37 MBq/0.1 ml). Sequential scintigrams were obtained using a gamma camera for 120 min. The injection site and the hind legs were massaged post-injection. Blood samples were obtained at 5, 15, 30, 60, 90 and 120 min. Two of the rabbits were killed after 2 h. Their organs were weighed and tissue specimens were obtained, weighed and counted against a standard using a gamma counter. The lymph channels and the lymph nodes were well visualized on the scintigrams. The background activity was very low, making interpretation easier. About 30% of the injected dose migrated from the injection site by 2 h. The mean popliteal lymph node uptake was 5.71 +/- 4.62% per gram of tissue. The lymph node to other tissue concentration ratios were very high, ranging from 63:1 for the kidneys to 1099:1 for the heart. We conclude that 99Tcm-HIG is a promising new agent for the visualization of the lymphatic system due to its easy labelling procedure, the stability of the label, its widespread availability and good image quality. It may potentially be useful in detecting and evaluating inflammatory lymph nodes.

Scintigraphic visualization of acute pancreatitis in cats with 99Tcm-citrate.

The accumulation of 99Tcm-citrate in abscesses and arthritic joints has previously been demonstrated. In the present study its usefulness was evaluated in 10 mongrel cats with acute pancreatitis induced by metronidazole infusion. Five cats, used as normal controls, were given physiological saline. All 15 cats were injected intravenously with 185 MBq 99Tcm-citrate. Scintigrams were obtained at 30 min intervals up to 2 h after injection by a gamma camera. The cats were then sacrificed. Tissue samples were obtained, weighed and counted against a 1/1000 dilution of the injected dose. In normal cats the kidneys and the urinary bladder were the only organs that could be visualized. In acute pancreatitis the pancreas and its exudate in the abdomen were also evident on the images. The pancreas to other neighbouring organ ratios increased 2.3-9.7 fold in acute pancreatitis compared to normal controls. Gross autoradiograms showed localization of radioactivity in the lobules of pancreas. Histologic examination of sections of pancreas demonstrated oedema, haemorrhage and some fat necrosis in pancreatic parenchyma, confirming acute pancreatitis. In conclusion, it has been demonstrated that acute pancreatitis in cats can be visualized scintigraphically with 99Tcm-citrate.

Scintigraphic imaging of uveal melanoma with 99Tcm-glutathione.

A prospective study with a new tumour-seeking agent, 99Tcm-glutathione (GSH), was performed on 17 patients with choroidal melanoma. Planar and SPECT images using 99Tcm-GSH clearly demonstrated melanotic melanoma but failed to show amelonotic melanomas. Following confirmation of our results by concurrent ultrasonography and magnetic resonance imaging or computed tomography, patients were managed by either 125I plaque brachytherapy, diode laser transpupillary thermotherapy or enucleation depending on the site and location. In combination with other diagnostic tests, 99Tcm-GSH scintigraphy may play a role in the detection of uveal melanoma and its possible distant metastases.

Detection of inflammatory lymph nodes in rabbits by 99mTc-HIG lymphoscintigraphy.

Tc-Human immunoglobulin G ( Tc-HIG) is a well-known radiopharmaceutical for the evaluation of inflammatory lesions. Recently, it has been demonstrated as a new agent for the visualization of the lymphatic system by our group. Our aim was to investigate the feasibility of detection of inflammatory lymph nodes by Tc-HIG lymphoscintigraphy. Ten adult New Zealand rabbits were used as group A. In a baseline study, 37 MBq Tc-HIG (0.1 ml) was injected into both hind legs of the rabbits, and sequential posterior gamma imaging with the rabbits lying prone was performed at 5, 15, 30, 60, 90 and 120 min using a single-headed gamma camera (Toshiba GCA G01 E). One week later, microorganisms ( ) were injected in a volume of 0.1 ml intradermally into the web space between the second and third toes in the bilateral hind legs of each rabbit in order to obtain inflammation in the popliteal lymph nodes. After 4 days, 37 MBq Tc-HIG (0.1 ml) was injected into the hind legs of the rabbits bilaterally, and sequential posterior gamma imaging was performed as described above (second study). Another group of 10 adult New Zealand rabbits (group B) was injected with the same microorganisms in the right hind legs only. After 4 days, scintigraphic imaging was carried out in the same way as described above (third study). Regions of interest were drawn over the injection sites and popliteal lymph nodes on each image for semiquantitative analysis. Count rates for each were calculated and a decay correction was applied. Time-activity curves were generated to show the percentage retention of radioactivity in each region. After the scintigraphic study, some of the group B rabbits were killed by intravenous injection of pentobarbitone (100-150 mg.kg, and both left and right lymph nodes were removed for microscopic examination. On the scintigrams, lymphatic channels and popliteal lymph nodes were visualized within 15 min. In the second study, bilateral popliteal lymph nodes were visualized more clearly than in the baseline study. The right popliteal lymph nodes of the rabbits were more clearly visualized in the third study. Semiquantitative analysis showed a higher percentage uptake of radioactivity in the right compared to the left popliteal lymph nodes in group B rabbits. Microscopic examination of the tissue sections demonstrated inflammation in the right lymph nodes of group B rabbits. In this preliminary study, it was found that Tc-HIG is a new promising agent for the demonstration and evaluation of inflammatory lymph nodes.