Use of bandage contact lenses for treatment of spontaneous chronic corneal epithelial defects in dogs.

OBJECTIVE: To determine whether polyxylon bandage contact lenses influence healing time and ocular comfort in the management of spontaneous chronic corneal epithelial defects in dogs. METHODS: Twenty dogs with spontaneous chronic corneal epithelial defects were included. All dogs were treated by debridement under topical anaesthesia at the first presentation. Ten dogs were assigned to the study group (application of a polyxylon bandage contact lens), and the remaining ten served as a control group. The healing time and ocular (dis)comfort were evaluated by assessment of the clinical findings and an owner-based questionnaire. All dogs received the same topical and systemic medication. RESULTS: Healing time for dogs in the study population was significantly shorter (mean 14±0 days) than for dogs in the control group (mean 36±17 days; P=0·005). The spontaneous chronic corneal epithelial defects had completely healed at the first recheck in all dogs with a polyxylon bandage contact lens. The duration of blepharospasm following debridement was significantly shorter in the study population (mean 4±4 days) than in the control group (mean 30±20 days; P=0·001). CLINICAL SIGNIFICANCE: The use of polyxylon bandage contact lenses is beneficial in the management of spontaneous chronic corneal epithelial defects.

RP-HPLC separation of the diastereomers of technetium-99m labelled tropanes and identity confirmation using radio-LC-MS.

99mTc-TRODAT-1 (technetium(V)-oxo-2-[[2-[[[3-(4-chlorophenyl)-8-methyl-8-azabicyclo[3.2.1]oct-2-yl]methyl](2-mercaptoethyl)amino]ethyl]]amino]-ethanethiolato(3-)) and 99mTc-TRODAT-M, the 4-methylphenyl derivative of 99mTc-TRODAT-1, are at this moment being evaluated in clinical trials as imaging agents for the central nervous dopamine transporter system. Both compounds are formed as a mixture of two major diastereomers. As the tracer concentration in preparations for clinical investigations is very low (30-150 pmol/ml), identification of these 99mTc-complexes was, up to now, carried out indirectly using X-ray diffraction analysis of the corresponding rhenium complexes which can be synthesized in gram amounts. In this study, we developed a convenient and practical reversed phase HPLC-method for purification and isolation of the respective diastereomers of 99mTc-TRODAT-1 and three of its derivatives using mixtures of solvents which are compatible with biological studies, i.e. aqueous buffers and ethanol. Furthermore, direct identity confirmation of the 99mTc-complexes using radio-LC-MS was successfully elaborated.

An efficient HPLC method for the analysis of isomeric purity of technetium-99m-exametazime and identity confirmation using LC-MS.

99mTc-exametazime (99mTc-d,l-HMPAO, 99mTc-d,l-hexamethylpropyleneamine oxime) is a neutral rather unstable complex of short-lived 99mTc (t(1/2)=6 h) with the d,l-isomer (mixture of D,D- and L,L-isomers) of a bis-amine bis-oxime tetraligand. It is widely used for measurement of regional cerebral perfusion in nuclear medicine. The meso-isomer (D,L-form) should not be present in a preparation as it is not retained in brain and thus does not provide clinically useful information. Meso-HMPAO is removed from the ligand during the synthesis procedure by repeated recrystallization, but can still be present as impurity in d,l-isomer. Due to the lack of a suitable chromatographic method for analysis of the isomeric purity of 99mTc-exametazime preparations, United States Pharmacopoeia 25 (USP 25) prescribes a biological test in rats for quality control purpose. In this study, we developed a suitable high-performance liquid chromatography (HPLC) method which allows to demonstrate the relative amounts of d,l- and meso-isomer in 99mTc-exametazime and so obviates the need for a biodistribution test in animals as part of the quality control. Due to the low concentrations in which 99mTc-d,l-HMPAO is obtained (typically 2-6 ng/ml), confirmation of the identity of 99mTc-d,l-HMPAO in the monograph of the European Pharmacopoeia is now performed only indirectly by TLC and assessment of its retention time on RP-HPLC. To investigate the potential of radio-LC-MS for assessment of the identity of 99mTc-exametazime, 99mTc-d,l-HMPAO and 99mTc-meso-HMPAO prepared using a Tc-rich eluate were analyzed using a radio-LC-MS system equipped with a time-of-flight mass spectrometer with electrospray ionization. The main peak in the radiometric channel coincided with the molecular ion mass of 99mTc-d,l-HMPAO in the mass spectrometer channel and the measured accurate mass differed only by 0.26 ppm from the theoretical mass. The identity of 99mTc-meso-HMPAO was also confirmed. Thus, radio-LC-MS allowed to obtain strong evidence for the structure of 99mTc-d,l-HMPAO and 99mTc-meso-HMPAO at nanomolar concentration. It is concluded that radio-LC-MS can become a sensitive aid in quality control of "no carrier added" radiopharmaceutical preparations.

Evaluation of 99mTc-MAMA-chrysamine G as an in vivo probe for amyloidosis.

To date, systemic amyloidosis is diagnosed histologically using Congo red staining or in vivo using iodine-123 labelled serum amyloid P component (123I-SAP) scintigraphy. We developed 99mTc-MAMA-CG, a 99mTc-labelled derivative of the lipophilic Congo red analogue chrysamine G (CG), as a possible alternative to 123I-SAP. In vivo 99mTc-MAMA-CG scintigraphy, performed in chickens with spontaneous joint amyloidosis, resulted as soon as 10 min after injection in scintigraphic images showing uptake of activity in amyloid-loaded organs (liver, joints). One of these chickens was studied also with 123I-SAP resulting in scintigraphic images revealing 123I-SAP binding to amyloid deposits in the liver. However, up to 11 h after injection no radioactivity was visible in the amyloid positive joints. In vitro autoradiography, performed on sections of chicken joints with Enterococcus faecalis induced amyloid arthropathy (chjAA), demonstrated the failure of 99mTc-MAMA-CG to bind significantly to amyloid deposits in the presence of 10 microM Congo red The specificity of 99mTc-MAMA-CG localisation was also established by the absence of 99mTc-MAMA-CG binding in non-amyloidotic organs in vitro and in vivo. 99mTc-MAMA-CG did not show any sign of acute toxicity. These findings establish the usefulness of 99mTc-MAMA-CG as a non-invasive in vivo diagnostic probe in chickens with amyloid arthropathy and suggest that it may also be applicable to human amyloidosis.

In vitro affinity of 99Tcm-labelled N2S2 conjugates of chrysamine G for amyloid deposits of systemic amyloidosis.

To date, systemic amyloidosis is diagnosed histologically in vitro using Congo red staining or in vivo using iodine-123 serum amyloid P component (123I-SAP) scintigraphy. 99Tcm-labelled derivatives of chrysamine G (CG), a lipophilic analogue of Congo red, were synthesized as potential tracer agents for direct and quantitative scintigraphic evaluation of amyloid deposits. To determine the affinity of 99Tcm-MAMA-CG, 99Tcm-Me4MAMA-CG and 99Tcm-MAMA-CG diethyl ester for amyloid, in vitro autoradiography was performed on sections of human kidney biopsy cylinders from kidneys with amyloid deposits (types AA, Alambda and Akappa) or control kidney tissue after incubation with the respective tracer agents. The binding of 99Tcm-MAMA-CG and its tetramethyl derivative was higher to kidney biopsy material with amyloid deposits of the AA, Alambda or Akappa type compared with control kidney tissue. This higher binding was prevented by the presence of 10 microM Congo red in the incubation medium. The diethyl ester of 9Tcm-MAMA-CG did not demonstrate increased binding to Congo red-positive kidney tissue. In conclusion, 99Tcm-MAMA-CG and 99Tcm-Me4MAMA-CG localize specifically to amyloid deposits in human kidney tissue, suggesting that these tracer agents may be applicable as specific targeting agents for diagnostic purposes in clinical amyloidosis.

The effect of intravenous medetomidine on pupil size and intraocular pressure in normotensive dogs.

Medetomidine, a highly specific alpha-2 adrenergic agonist, has been demonstrated to lower intraocular pressure (IOP) in rabbits and cats when applied topically. The purpose of this study was to assess the influence of intravenously injected medetomidine on the pupil size (PS) and the IOP of non glaucomatous dogs. IOP was measured by applanation tonometry and PS was measured using Jameson calipers at t=0 (or time of IV injection of medetomidine (Domitor; Orion) at the dose of 1500 microg/m2 body surface area) and again after 5 minutes (t=5). The IV administration of medetomidine caused miosis in all 14 dogs. The mean PS decreased from 9.0 to 4.0 mm (p<0.001). The IOP was lowered in 10 dogs and in 4 dogs there was a rise in IOP. The mean IOP (mmHg) decreased from 22 to 21 (p>0.2). The data presented above confirm that medetomidine at a dose of 1500 microg/m2 body surface area produces miosis in non glaucomatous dogs, without influencing the IOP.

Measurement of plasma volume using 99Tc(m)-labelled DMP-HSA.

Red cell volume (RCV) and plasma volume (PV) measurements are performed routinely in nuclear medicine departments to diagnose a number of haematological disorders. Currently, 125I-HSA is used as a plasma tracer and 99Tcm-labelled red cells to determine red cell volume. 125I-HSA is not always readily available, leading to inconvenience for patients and medical practitioners. Due to the availability of 99Tcm in nuclear medicine departments, the use of albumin labelled with 99Tcm was investigated. A new 99Tcm-human serum albumin labelling kit (99Tcm-DMP-HSA) was developed by Verbeke and supplied for use in this study. The main aim of the study was to investigate the use of 99Tcm-DMP-HSA for PV determination. Secondly, the feasibility to determine red cell and plasma volume simultaneously using 99Tcm as radionuclide in both instances was investigated. Fourteen healthy volunteers were enrolled in the dual-phase study. During the first study, 99Tcm-DMP-HSA was used as tracer to calculate PV (PV1a) after intravenous administration. Subsequently, 99Tcm-labelled red cells were administered and the PV (PV1b) and RCV (RCV1) were calculated. The second study was repeated within 2 weeks using the conventional method. 125I-HSA and 99Tcm-labelled red cells were administered simultaneously. The PV (PV2) and RCV (RCV2) were calculated. We found that the redistribution of 99Tcm-DMP-HSA is faster than that of 125I-HSA; therefore, the plasma counts obtained at different times were back-extrapolated to time zero for plasma volume calculations. The mean values for the different calculated PVs were 2964+/-470 ml for PV1a, 3006+/-623 ml for PV1b and 3001+/-530 ml for PV2, the reference PV. The confidence intervals indicate no significant differences between plasma volumes PV1a and PV2 and plasma volumes PV1a and PV1b. The mean calculated RCV1 was 2130+/-322 ml; that of RCV2 was 2128+/-353 ml. The difference between RCV1 and RCV2 was not significant. Our results indicate that 99Tcm-DMP-HSA could be used for plasma volume calculation. Red cell and plasma volumes can be calculated simultaneously using 99Tcm as radionuclide in both cases.

Synthesis and biological evaluation of the four isomers of technetium-99m labeled ethylenecysteamine cysteine ((99m)Tc-ECC), the mono-acid derivative of (99m)tc-L,L-ethylenedicysteine.

A few years ago (99m)Tc-ethylenedicysteine ((99m)Tc-L,L-EC) had been proposed as an interesting substitute for technetium-99m labeled mercaptoacetyltriglycine (MAG3) as renal function tracer agent. It possesses in its structure two carboxylate functions and is in this respect different from other renal tracers such as (99m)Tc-N, N'-bis-(mercaptoacetyl)-2,3-diaminopropionate ((99m)Tc-CO(2)DADS), (99m)Tc-MAG3, and Hippuran, which have only one carboxylic group. To study whether both carboxylic acid groups of (99m)Tc-L,L-EC contribute to the efficient renal handling of this compound we synthesized and biologically evaluated the technetium-99m labeled isomers of L- and D-ethylenecysteamine cysteine (ECC), the mono-acid derivative of (99m)Tc-L,L-EC. Labeling of L-ECC or D-ECC with (99m)Tc using a direct or exchange labeling method yields for each of them two diastereomeric (99m)Tc complexes (A and B, in the order of elution during reversed phase high performance liquid chromatography) in relative amounts depending on the pH during labeling. In mice, all four isomers of (99m)Tc-ECC (LA, LB, DA, and DB) are cleared rapidly from the blood, mainly by the renal system. The isomers LB and DB show the most efficient renal handling, but none of the mono-acid derivatives has a urinary excretion rate as high as that of (99m)Tc-L,L-EC. The renal handling of the isomers of (99m)Tc-ECC is partly due to tubular secretion because the urinary excretion of these compounds is significantly lower in mice pretreated with probenecid. In the baboon, isomers DA and DB show a plasma clearance comparable to that of (99m)Tc-L,L-EC. The plasma clearance of isomers LA and LB is lower but still comparable to or higher than that of (99m)Tc-MAG3. In a human volunteer, isomer DB shows a plasma clearance rate only slightly lower than that of (99m)Tc-L,L-EC. Thus, it appears that the presence of one carboxylate in (99m)Tc-EC-like compounds can be sufficient for efficient renal handling. However, it is also evident that the configuration at the chiral carbon atom and the orientation of the oxotechnetium core determine in a significant way the biological characteristics.

Reduction of contamination risks during clinical studies with technegas.

During patient studies with the Technegas equipment in our department, we regularly detected small to considerable contaminations of the operator and in the area surrounding the apparatus. These contaminations were found to be of different origin: residual activity in the tubing from the apparatus to the patient which diffuses after deconnection, residual activity and small particles of the destroyed carbon crucible in the apparatus which are dispersed during opening of the door of the gas preparation chamber, leakage at the patient site during studies in uncooperative patients and a dysfunctioning valve inside the apparatus. To reduce the contamination risks, we made some adaptations to the apparatus. In the first place, the dysfunctioning valve was replaced. In addition, a powerful air exhaust pump with an efficient filter was installed. It was connected with (1) a newly installed transparent box in front of the door of the gas preparation chamber, (2) a dome on a flexible arm which can be positioned above the patient's face during the examination and (3) a nipple on which the mouthpiece can be placed after the study. After these adaptations, a study showed the absence of measurable contamination on the clothing of the personnel handling the apparatus. Occasionally, considerable contamination was still measured on the gloves worn during filling of the carbon boat with generator eluate, but only small contaminations (up to 9.25 kBq) were measured on the mouthmask worn by the operator during administration of the Technegas. This results in a maximum effective dose equivalent from activity deposited in the lungs of 0.008 microSv per study. The total body dose of the operator from external radiation for one Technegas examination was determined to be 2 microSv. The highest dose rate was measured during filling of the crucible (0.2 mSv/h).

99mTc-MAMA-chrysamine G, a probe for beta-amyloid protein of Alzheimer's disease.

Chrysamine G (CG), an analogue of Congo red, is known to bind in vitro to the beta-amyloid protein (Abeta 10-43) and to homogenates of several regions of the brain of Alzheimer's disease (AD) patients. We synthesised a conjugate of 2-(acetamido)-CG with a bis-S-trityl protected monoamide-monoaminedithiol (MAMA-Tr(2)) tetraligand, which was efficiently deprotected and labelled with a 75% yield with technetium-99m, to obtain (99m)Tc-MAMA-CG. In mice, (99m)Tc-MAMA-CG was cleared mainly by the hepatobiliary system, resulting in a fast blood clearance. Brain uptake of (99m)Tc-MAMA-CG was low. Co-injection with the blood pool tracer iodine-125 human serum albumin ((125)I-HSA) demonstrated a brain/blood activity ratio for (99m)Tc-MAMA-CG that was significantly higher than that for (125)I-HSA (t test for dependent samples, P<0.02), indicating the ability of (99m)Tc-MAMA-CG to cross the blood-brain barrier. In vitro autoradiography demonstrated pronounced binding of (99m)Tc-MAMA-CG to beta-amyloid deposits in autopsy sections of the parietal and occipital cortex of an AD patient as compared with controls. Adding 10 microM Congo red during incubation displaced the binding of (99m)Tc-MAMA-CG. Congo red staining and autoradiography identified the same lesions. (99m)Tc-MAMA-CG seems to bind selectively to beta-amyloid deposition in human brain parenchyma and blood vessels in vitro and thus might be a lead compound for further development of a useful tracer agent for the in vivo diagnosis of Alzheimer's disease.

Development of a lyophilized kit formulation for labeling of DNA probes with 99mTc.

DNA fragments such as oligodeoxynucleotides (ODNs) are under investigation for a possible utilization in nuclear medicine. Until now, experiments on 99mTc-labeled ODNs in vitro or in vivo have required the application of time-consuming procedures to obtain and control the purity of the radiolabeled compound. A lyophilized labeling kit would ease and improve the reproducibility in further investigations with this class of promising biomolecules; therefore a study was initiated to evaluate the suitability of conjugates of ODNs and a bifunctional chelating agent to be part of lyophilized kit formulations. We report here the development of the first kit for one-step labeling of oligonucleotides with 99mTc. The formulation comprises 250-500 pmol S-benzoyl-mercaptoacetyldiglycine (MAG2)-ODN phosphorothioate conjugate, 5 mg potassium sodium tartrate tetrahydrate and 100 microg stannous chloride dihydrate in a lyophilized kit. Labeling yields above 90% were reproducibly achieved after addition of 0.1-1 GBq pertechnetate and subsequent heating in a boiling water bath. Once formed, the 99mTc-MAG2-ODN complexes were stable for at least 24 h. The shelf life of the kits is at least 10 weeks when stored protected from light at room temperature, but even kits stored at 40 degrees C gave labeling yields above 90% after 10 weeks.

Biodistribution and absorbed radiation dose estimates of 99mTc-labeled dimercaptopropionyl human serum albumin.

UNLABELLED: The use of 99mTc-labeled red blood cells (RBC) for the evaluation of left ventricular function using equilibrium-gated blood-pool imaging suffers from several problems and potential risks. In this study, we estimated the absorbed radiation dose of 99mTc-labeled dimercaptopropionyl human serum albumin (DMP-HSA) as a potential alternative to 99mTc-RBC. METHODS: After the administration of 99mTc-DMP-HSA, whole-body imaging was performed up to 48 h after injection in five volunteers. The heart contents, liver and remainder of the body were used as source organs. Multicompartment modeling of the biodistribution was performed and absorbed radiation dose estimates for 99mTc-DMP-HSA were obtained using the Medical Internal Radiation Dose (MIRD) calculation. RESULTS: Residence times of 0.62 and 0.43 h were obtained for the heart contents and liver, respectively. Radiation dose estimates yielded an effective dose of 0.0055 mSv/MBq. CONCLUSION: 99MTC-DMP-HSA yielded absorbed radiation doses comparable with those of 99mTc-RBC. Therefore, the radiation properties of 99mTc-DMP-HSA are such that it can be used for clinical diagnostic studies.

Persistent hyperplastic tunica vasculosa lentis and persistent hyaloid artery in a 2-year-old basset hound.

A 2-year-old Basset hound with unilateral persistent hyperplastic tunica vasculosa lentis and primary vitreous is described. There was leukokoria in the left eye, caused by bluish-white polar densities and haemorrhagic discoloration in the nucleus of the lens. Ultrasonographic examination of the eye revealed a small hyperechoic lens and a hyperechoic linear soft-tissue strand extending from the posterior pole of the lens through the optic disc. Power Doppler imaging revealed blood flow in this persistent hyaloid artery, but 5 months later, immediately prior to surgery, there was no flow in this artery. Ultrasonography, including Power Doppler imaging, was useful to confirm the diagnosis and to estimate the complications of surgery, especially the risk of vitreal haemorrhage after cutting of the persistent hyaloid artery. Comparison of the ultrasonographic images with those obtained by computed tomography suggested that the resolution of computed tomography is inadequate for identifying structures as fine as a persistent hyaloid artery.

3'-99mTc-labeling and biodistribution of a CAPL antisense oligodeoxynucleotide.

CAPL is a cancer-related gene shown to be overexpressed during tumor metastasis formation. A CAPL antisense oligodeoxynucleotide (ODN), GX-1, and a random control ODN (CTRL1) were 3'-conjugated to MAG3, labeled with technetium-99m, purified, and the biodistribution of the radiolabeled conjugates in normal mice was studied. A 99mTc-MAG3-GX-1 complex of >97% radiochemical purity was obtained and the product was stable for >6 h as determined by reversed phase high performance liquid chromatography (HPLC). Biodistribution studies of the 99mTc-MAG3-ODNs in groups of four normal mice, sacrificed 5 min and 60 min after injection, demonstrated that the radiolabeled ODNs were distributed in an unspecific manner. The excretion route was mainly urinary. At 60 min, 55.2% of the injected dose of 99mTc-MAG3-GX-1 and 72.4% of 99mTc-MAG3-CTRL1 was found in the urine. This finding is clearly different from previously reported data on tritiated 20-mer phosphodiester ODNs, as well as the unconjugated 99mTc-MAG3 chelate, suggesting that 99mTc-MAG3 coupled to the 3'-end of ODNs has an influence on the biodistribution of the oligo, and possibly has a protective function to enzymatic degradation in vivo.

Ultrasonographic diagnosis of persistent hyperplastic tunica vasculosa lentis/persistent hyperplastic primary vitreous in two dogs.

Ultrasonography was performed on a Basset hound and a Doberman clinically suspected of persistent hyperplastic tunica vasculosa lentis/persistent hyperplastic primary vitreous. In both dogs, hyperechoic lenses with a triangular-shaped echodense structure retrolentally were visible, and a very thin hyperechoic strand was seen penetrating the anechoic vitreous from this retrolental tissue to the area of the optic nerve. Using color Doppler imaging, blood flow was evident in parts of the retina in both dogs. With power Doppler imaging there was blood flow in the lens and hyperechoic strand of the Basset hound on the first examination; whereas, on re-examination 5 months later, this was not found. Other abnormalities, such as retinal detachment, endophthalmitis, vitreous hemorrhage, microphthalmia, and posterior neoplasia could be excluded.

Importance of the two ester functions for the brain retention of 99mTc-labelled ethylene dicysteine diethyl ester (99mTc-ECD).

99mTc-ethylene dicysteine diethyl ester (99mTc-L,L-ECD) is a neutral lipophilic tracer agent that crosses the blood-brain barrier and is retained in the brain of primates following enzymatic hydrolysis of one of the ester functions to the ionized mono-ester, mono-acid metabolite. Up to now, it is not clear whether the second ethylcarboxylate group is essential for brain uptake and retention. Therefore, we have synthesized and studied two derivatives of 99mTc-L,L-ECD that contain only one ethylcarboxylate function, namely 99mTc-labelled L- and D-ethylene cysteamine cysteine ethyl ester (99mTc-ECCE). Direct labelling of L- or D-ECCE at neutral pH and room temperature resulted for each of them in the formation of two probably diastereomeric 99mTc-complexes in a 1:1 ratio. This means that four different isomers could be isolated. The 99mTc-labelled complexes formed after labelling ECCE (A and B, in order of elution during HPLC) are slightly less lipophilic than 99mTc-L,L-ECD. In mice, all four isomers show a low brain activity at 10 min post injection (p.i.), approximately 0.1% to 0.3% of the injected dose versus 0.9% for 99mTc-L,L-ECD. The clearance from the blood is comparable with (isomers LA and LB) or slower (isomers DA and DB) than that of 99mTc-L,L-ECD. Isomers LA and DA show high liver uptake and rapid excretion to the intestines. Both 99mTc-ECCE-LB and 99mTc-ECCE-DB, the most lipophilic isomers, are cleared from the blood mainly by the kidneys and excreted more efficiently to the urine. 99mTc-ECCE-LB is characterized by a surprisingly high heart uptake (about 1.5% of i.d.) at 10 min p.i. versus 1.0% for 99mTc-methoxyisobutylisonitrile (99mTc-MIBI) and 0.2-0.3% for the other isomers, but also lung uptake is relatively high. In the baboon, brain and heart uptake of isomers LA and LB of 99mTc-ECCE were negligible. Activity concentrated mostly in the hepatobiliary system for isomer LA and in the renal system for isomer LB. The results indicate a clear difference in biological behaviour between 99mTc-L,L-ECD and the four isomers of 99mTc-ECCE. This shows that the presence of both ester functions in 99mTc-L,L-ECD is an essential structural requirement for its brain uptake and retention in primates.

Hybridization of a 99Tcm-labelled oligodeoxynucleotide to CAPL RNA.

Oligodeoxynucleotides (ODNs) labelled with an appropriate radionuclide could provide a means to identify serious diseases early on and thereby help initiate treatment at a very early phase. Regardless of important issues like in-vivo stability and membrane passage, the key issue for the oligonucleotide approach is the ability of the radiolabelled ODN to hybridize to the target mRNA. The secondary structure of mRNA does not permit all complementary ODNs to hybridize and a careful selection of the probe with consecutive testing is therefore necessary. This study was initiated to demonstrate hybridization of a 99Tcm-labelled 20-mer ODN to RNA of CAPL (S100A4), a gene reported to be overexpressed in metastatic cancers like breast carcinoma and osteosarcoma. The phosphodiester ODN GX-1 (antisense) and two control sequences (scrambled and random) were conjugated to the bifunctional chelating agent S-benzoyl-mercaptoacetyltriglycine (S-benzoyl-MAG3) and labelled with 99Tcm. The radiolabelled ODNs were purified on a C18 mini-column and characterized on a reverse-phase HPLC system. The radio-chemical purity was > 90% and the product was stable for > 6 h in aqueous medium. The hydrization properties of unlabelled, 32P-labelled and 99Tcm-labelled ODNs to transcribed RNA were studied using polyacrylamide gel electrophoresis (PAGE). Direct hybridization of GX-1 to transcribed RNA was demonstrated. A 50-fold excess of unlabelled ODN over transcribed RNA caused a near to complete consumption of RNA by RNase H activation. In 1:1 proportions of radiolabelled (32P and 99Tcm) ODNs to RNA, only radiolabelled GX-1 was found to hybridize to RNA in a PAGE system. The radiolabelled control ODNs did not show signs of hybridization. This study demonstrates that 3'-99Tcm-labelling of ODNs does not interfere with the hybridization properties of the ODNs in solution, making 99Tcm-labelling an attractive procedure for the future development of antisense technology in imaging.

Comparison of modified technetium-99m albumin and technetium-99m red blood cells for equilibrium ventriculography.

UNLABELLED: A newly developed modified form of 99mTc-labeled human serum albumin reconstituted from a kit (99mTc-dimercaptopropionyl-human serum albumin; 99mTc-DMP-HSA) was prospectively compared to 99mTc-labeled red blood cells (RBC) in patients referred for equilibrium radionuclide ventriculography at rest to evaluate its potential use as a blood-pool imaging agent. METHODS: A Paired comparison between 99mTc-DMP-HSA and either in vitro or in vivo 99mTc-labeled RBC was performed within 2 days in 20 patients'. For each study, two sets of images were acquired, starting at 15 min and 180 min postinjection, respectively. Each set consisted of a gated blood-pool cardiac study and a planar static image centered on the patient's thorax. All data were processed by two independent observers. Early and late postinjection parameters were calculated: ejection fraction (EF) value, activity within the main organs surrounding the left ventricle (LV), ratio of activity between the LV and these surrounding organs for each study separately, and temporal (late/early) evolution of the intraorgan activities and of the LV/organ ratios after decay correction. RESULTS: The images and the visual wall-motion analysis were of good quality with both agents in most patients, without significant image degradation at 180 min postinjection. Calculated EF values were highly comparable with the two tracers. Interobserver variability was 0.17% (RBC) and 1.08% (DMP-HSA) for the early EF value (EF1), and 0.62% (RBC) and 0.27% (DMP-HSA) for the late EF (EF2). Mean difference between EF2 and EF1 was 0.74% (Observer 1) and 0.28% (Observer 2) for 99mTc-RBC, and -2.88% (Observer 1) and -2.07% (Observer 2) for 99mTc-DMP-HSA. When comparing 99mTc-DMP-HSA to 99mTc-RBC the mean difference was 1.27% (Observer 1) and 0.36% (Observer 2) for EF1, and -2.35% (Observer 1) and -1.99% (Observer 2) for EF2. Also, the biodistribution and temporal evolution of the organ repartition of both compounds were stable and similar, with values of late/early activity ratios very close to one for all the studied organs [mean intraorgan ratio: 0.946 for 99mTc-RBC (range: 0.881-1.086) and 0.979 for 99mTc-DMP-HSA (range: 0.914-1.141); mean late/early LV/organ ratio: 0.964 for 99mTc-RBC (range: 0.919-1.016) and 0.967 for 99mTc-DMP-HSA (range: 0.912-1.035)]. CONCLUSION: Paired comparison of kit-prepared 99mTc-DMP-HSA to 99mTc-labeled RBC demonstrated that both agents were very closely related regarding as well the calculated EF value as the in vivo stability up to more than 3 hr postinjection. Technetium-99m-DMP-HSA may constitute a practical and useful replacement for 99mTc-labeled RBC.

Development and evaluation of a kit formulation for the preparation of 99mTc-DMP-HSA, a new tracer agent for radionuclide ventriculography.

This study presents the development of a kit formulation for the preparation of 99mTc-DMP-HSA, followed by a comparison of such kit-prepared 99mTc-DMP-HSA to 99mTc-RBCs in a volunteer. Reconstitution of the labeling kits with up to 5.55 GBq 99mTc afforded 99mTc-DMP-HSA preparations with a > 95% radiochemical purity for up to 8 h. Only minor differences were observed in the global distribution of both tracer agents, whereas the calculated ejection fractions were almost identical. The effective dose equivalent of 99mTc-DMP-HSA is 8.68 microSv/MBq.

Clinical evaluation of technetium-99m-L,L-ethylenedicysteine in patients with chronic renal failure.

UNLABELLED: Technetium-99m-L,L-ethylenedicysteine (99mTc-L,L-EC), a new renal radiopharmaceutical, has been shown to have similar excretion characteristics but a higher plasma clearance than 99mTc-mercaptoacetyltriglycine (99mTc-MAG3) in normal volunteers and patients with obstructive nephropathy. This study evaluated 99mTc-L,L-EC in patients with chronic renal failure. METHODS: The clearance of 99mTc-L,L-EC was compared with that of 125l-hippuran in 26 patients with varying degrees of chronic renal impairment (serum creatine 168-1163 mumol/liter). All 26 patients also were imaged with 99mTc-L,L-EC (70-80 MBq). Fifteen patients had further imaging with 99mTc-MAG3 (100 MBq) the following day. RESULTS: A subjective analysis of the 99mTc-L,L-EC images revealed that all were of acceptable quality regardless of creatinine level. In the 15 patients who were imaged with both 99mTc-L,L-EC and 99mTc-MAG3, general image quality and target-to-background ratios were similar. Time-activity curves and mean parenchymal transit times obtained with the two agents were almost identical. Plasma clearance values (mean +/- s.d.) of 99mTc-L,L-EC and 125l-hippuran were 81 +/- 68 ml/min and 114 +/- 104 ml/min, respectively. Mean 99mTc-L,L-EC clearance was 71% of the mean 125l-hippuran value. CONCLUSION: Technetium-99m-L,L-EC provides equally high-quality images to 99mTc-MAG3 in patients with chronic renal failure. Technetium-99m-L,L-EC clearance more closely resembles that of hippuran than does 99mTc-MAG3 clearance. These features together with its ease of preparation make 99mTc-L,L-EC an attractive alternative to 99mTc-MAG3 in patients with chronic renal failure.