Biomedical subjects

Chris Tsopelas

Publications and source records attributed to Chris Tsopelas.

15 recordsLinked to original sources

PubMed2006

99mTc-Evans blue dye for mapping contiguous lymph node sequences and discriminating the sentinel lymph node in an ovine model.

BACKGROUND: The aim of this study was to investigate the potential of (99m)Tc-Evans blue for discriminating the sentinel lymph node in multitiered lymph node sequences by using an ovine model. (99m)Tc-Evans blue is an agent that has both radioactive and color signals in a single dose. Previous studies in smaller animal models suggested that this agent could have advantages over the dual-injection technique of radiocolloid/blue dye. METHODS: Doses of (99m)Tc-Evans blue ( approximately 21 MBq) containing Evans blue dye (approximately 4 mg) were administered to the hind limbs or fore limbs of sheep to map the lymphatic drainage patterns, validate its ability to identify the sentinel lymph node, and examine the reproducibility of the technique. The study protocol was repeated with (99m)Tc-antimony trisulfide colloid and Patent Blue V dye. After the operative exposure, lymph nodes were identified with the gamma probe and then excised and analyzed for radioactivity (percentage of injected dose) and blue color. RESULTS: After the administration of (99m)Tc-Evans blue, all lymph nodes harvested (35 of 35) in either short chains or long basins were hot and blue. The sentinel lymph nodes concentrated more radioactivity than the second-tier nodes to the extent of 2:1 to 215:1. For radiocolloid/Patent Blue V, the ratios were lower, at 2:1 to 3:1. CONCLUSIONS: (99m)Tc-Evans blue was found to better discriminate the sentinel lymph node than (99m)Tc-antimony trisulfide colloid/Patent Blue V in variable multitier lymph node anatomy, and it is an agent that promises to have positive clinical applications.

Animals↗

PubMed2006

The tissue distribution of Evans blue dye in a sheep model of sentinel node biopsy.

BACKGROUND: Tc-Evans blue is a 'single dose' agent for lymphatic mapping combining radioactivity and blue dye for sentinel node identification. The mechanism and distribution of blue dye retention in the lymph node is not clearly understood. OBJECTIVE: To demonstrate the cellular distribution of Tc-Evans blue in sheep sentinel lymph nodes by measuring the radioactivity of different tissue components and correlating this with pathological examination. METHODS: Tc-Evans blue was used to identify sheep lymph nodes. Part of each node was sent for pathological examination including imprint cytology, and frozen and permanent section examination. Sections were examined without stains, with only red stains and conventional haematoxylin & eosin staining. The remaining nodal tissue was homogenized and components separated by enzymatic digestion and density gradient centrifugation. Fractions representing each tissue component were counted in a gamma counter and the distribution of Tc-Evans blue calculated. RESULTS: A dispersed population of blue staining cells was found. Their distribution, number and size indicated that they were histiocytes such as macrophages or antigen presenting cells. Radioactivity was distributed throughout the lymph node. Over 70% remained in the plasma, 19% in the leukocyte layer, and 10% was associated with erythrocytes and undigested tissue. CONCLUSION: The accumulation of radioactivity and blue colour in the lymph nodes indicates the mechanism of retention is a result of the binding interaction between Tc-Evans blue-protein and lymph node histiocytes including macrophages and antigen presenting cells.

Animals↗

PubMed2005

An improved (99m)Tc-aprotinin kit formulation: quality control analysis of radiotracer stability and cold kit shelf life.

(99m)Tc-aprotinin scintigraphy has been demonstrated to be a useful noninvasive imaging technique for amyloid deposits located in extraabdominal regions of patients. The aim of this study was to develop an improved aprotinin cold kit formulation, to validate the kit for long-term stability, as well as to assess the radiotracer stability by novel quality control methods. The aprotinin cold kit formulation of Trasylol, pyrophosphate (PYP)-chelated stannous reductant and an alkaline buffer, was dispensed into nitrogen-filled vials and aliquots frozen at -20 degrees C. After 0, 1, 2, 3 and 6 months of storage, three samples were reconstituted with 750-850 MBq of (99m)Tc-pertechnetate, followed by quality control analyses by paper chromatography methods at 25, 85 and 265 min postreconstitution (pr). Cation-exchange cartridge quality control methods were also investigated. The cold kits proved to be stable to long-term storage for up to 6 months, and the radiotracer was stable for at least 4 h pr. (99m)Tc-aprotinin was formed at greater than 95% efficiency at all time points tested with (99m)TcO2 present as the major impurity (1-4%) and (99m)Tc-pertechnetate and (99m)Tc-PYP present in trace amounts. An alternative, rapid, safe and reliable method was found in Oasis MCX-BSA-treated cartridges using saline as the eluting solution to assay for (99m)Tc-aprotinin.

Aprotinin↗

PubMed2005

Pharmacokinetics and normal scintigraphic appearance of 99mTc aprotinin.

OBJECTIVE: To confirm the pharmacokinetics and biodistribution of 99mTc aprotinin in normal volunteers and to determine the optimum time for scanning post-injection, prior to further investigations of 99mTc aprotinin as an imaging agent for amyloidosis. METHODS: Five patients (three men and two women, average age 49 years, age range 38-66 years) without a history of amyloidosis or any of the associated diseases, were included in this prospective study. Blood and urine were collected and images were performed of the whole body and wrists. CONCLUSIONS: Normal biodistribution of 99mTc aprotinin includes early cardiac and lung activity in the blood pool phase with subsequent hepatic activity and renal excretion with variable splenic activity. There is variable bowel uptake on later images. The best quality images were obtained 90 min post-intravenous administration, and this is likely to be the optimum time for clinical imaging.

Adult↗

PubMed2005

Use of [14C]-sodium bicarbonate/urea to measure physical activity induced increases in total energy expenditure in free-living healthy males.

The aim of this study was to evaluate the utility of the [(14)C]-sodium bicarbonate/urea technique to detect physical activity-induced increases in total energy expenditure in free-living healthy men. Thirteen healthy males aged 34.1 +/- 11.7 yrs with body mass index 24.1 +/- 3.1 kg/m(2) were studied on three separate occasions, during which [(14)C]-bicarbonate was infused over 48-hours and urine was collected during the second 24-hours. On three separate occasions and in random order, subjects either remained sedentary, or performed a bout of physical activity on an electro-magnetically braked cycle ergometer sufficient to increase energy expenditure by 7% or 11% above predicted sedentary total energy expenditure. Urine samples were analyzed to evaluate the amount of [(14)C]-bicarbonate incorporated into urinary urea, thereby reflecting the amount of CO(2) produced per day, and upon conversion, the number of kilojoules of energy expended in 24-hours. All 13 subjects successfully completed the two physical activity treatments and there were no adverse events. As measured by the [(14)C]-urea assay, mean total energy expenditure values were not significantly different between sedentary activity (17902 +/- 905 kJ/day), the physical activity treatment designed to increase TEE by 7% (17701 +/- 594 kJ/day) and the physical activity treatment designed to increase TEE by 11% (18538 +/- 485 kJ/day) (P=0.668). In conclusion, although the [(14)C]-sodium bicarbonate/urea technique was well tolerated and did not interfere with normal daily activities, it was not able to accurately measure physical activity-induced increases in EE in the range of 7-11% above predicted sedentary total energy expenditure.

Adolescent↗

PubMed2005

Characterization and quality control analysis of 99mTc-bicisate.

The aim of this study was to investigate the mini cartridge versus paper chromatography quality control methods for determining the radiochemical purity (RCP) of (99m)Tc-bicisate. The 4 methods that were compared with the manufacturer's method included Whatman 17 paper/ethyl acetate solvent, instant thin-layer chromatography (ITLC) silica gel paper/saline solvent, reverse-phase C18 mini cartridge/saline solvent, and strong anion exchange mini cartridge/water solvent. At 30 min after reconstitution, (99m)Tc-bicisate was formed at 97%-98% RCP as assayed by the paper and cartridge methods, and the strong anion exchange/water for injection (WFI) system slightly underestimated the percentage at 96%. A significantly lower RCP was obtained for the C18/saline method when a faster flow rate was used. The lipophilic complex moved with ethyl acetate on Whatman 17, was separated from origin impurities on ITLC silica gel/saline, and remained on the column with C18/saline. For strong anion exchange/WFI, components in the radioactive formulation are likely to have influenced the percentage of (99m)Tc-bicisate. The time disadvantage for ITLC silica gel/saline analysis made the method less than ideal. The C18 mini cartridge/saline method was found to be the simplest and fastest; a result was obtained in 2 min with use of a safe solvent of elution.

Chromatography↗

PubMed2003

Regenerating lizard tails: a new model for investigating lymphangiogenesis.

Impaired lymphatic drainage in human limbs causes the debilitating swelling termed lymphoedema. In mammals, known growth factors involved in the control of lymphangiogenesis (growth of new lymph vessels) are vascular endothelial growth factors-C and -D (VEGF-C/D). Here we characterize a model of lymphangiogenesis in which the tail of lizards is regenerated without becoming oedematous. Three weeks after the tail is shed (autotomy), there are a small number of large diameter lymphatic vessels in the regenerated tail. Thereafter, the number increases and the diameter decreases. A functional lymphatic network, as determined by lymphoscintigraphy, is established 6 wk after autotomy. The new network differs morphologically and functionally from that in original tails. This lymphatic regeneration is associated with an up-regulation of a reptilian homologue of the VEGF-C/D protein family (rVEGF-C/D), as determined by Western blot analysis using a human reactive VEGF-C polyclonal antibody. Regenerating lizard tails are potentially useful models for studying the molecular basis of lymphangiogenesis with a view to developing possible treatments for human lymphoedema.

Animals↗

PubMed2003

Effect of port composition on tumor cell adherence: an in vivo model.

INTRODUCTION: We have reported previously on an in vitro model to examine tumor cell adherence to metal and plastic laparoscopic ports and to port sites through which they had been passed. This demonstrated that increased numbers of tumor cells were found both on metal ports compared with plastic ports and on the port sites through which metal ports had passed. In this study, the in vivo adherence of such cells to ports and port sites was investigated. METHODS: LIM 1215 tumor cells were injected under direct vision into the pelvises of 16 30-kg female pigs (range, 15-70 x 106 cells). A total of 12 ports were inserted through each anterior abdominal wall (6 metal and 6 plastic), and these were either left in situ for 30 minutes (nondisplaced) or were removed twice and replaced through the original wound (displaced). RESULTS: Increasing the tumor cell inoculum resulted in increased deposition of tumor cells on both ports (P = 0.002) and on the port sites (P = 0.017). Significantly more tumor cells adhered to metal ports than to plastic ports (P = 0.04), although this failed to reach significance for the sites through which metal ports had been passed (P = 0.066). Although displacement of ports did not increase the number of tumor cells that adhered to ports (P = 0.45), this did result in more tumor cells being deposited on the port sites (P = 0.01). CONCLUSIONS: These data suggest that minimizing the number of tumor cells within the abdominal cavity, using plastic ports, and securing ports to prevent inadvertent displacement would be expected to reduce the number of tumor cells deposited in port sites during operative laparoscopy. This may be beneficial in reducing the incidence of port-site metastases after laparoscopic surgery for gastrointestinal malignancies.

Animals↗

PubMed2003

A simple and effective technique to reduce staff exposure during the preparation of radiopharmaceuticals.

OBJECTIVE: In the preparation of radiopharmaceuticals, staff receive considerable radiation exposure to the hands during withdrawal of activity from the elution vial, from a combination of the syringe and elution vial activities. In an attempt to reduce the radiation burden to the hands, a simple technique was developed that utilizes a modified lead pot lid and a syringe bearing a long needle with a sterile needle guide. METHODS: An elution vial in a lead pot with an attached syringe was secured at an angle of 45 degrees, simulating the action of withdrawing (99m)Tc-pertechnetate from the elution vial. The (99m)Tc activity ratio of vial to syringe was 20:1. A gamma- camera detector without collimator was positioned at the syringe plunger and count profiles were obtained after 10 min of data acquisition. The experiment was repeated using the same set-up with (a) the modified lid on the lead pot and (b) a cold syringe to determine the contribution of the radioactive syringe to the count profile. Each experiment was repeated at the vertical position, simulating the normal action of redispensing (99m)Tc activity into the elution vial. RESULTS: The modified lid reduced exposure from the elution vial, with a count reduction of >98% for both orientations. The contributions of vial radioactivity to the total count profile were 76% and 84% for vertical and 45 degrees orientations, respectively. The contributions of syringe activity were 24% and 16% for vertical and 45 degrees orientations, respectively. CONCLUSION: A reduction in the photon flux to the hands of up to 84% (with an associated reduction in hand dose) can be achieved by withdrawing activity through a modified lid on the lead pot housing the elution vial, without significantly altering normal work practices.

Humans↗

PubMed2002

Sentinel node biopsy and lymphoscintigraphy with a technetium 99m labeled blue dye in a rabbit model.

BACKGROUND: Lymphatic mapping for sentinel node biopsy in breast cancer and melanoma usually involves initial peritumoral injection of a radioisotope, gamma camera detection of the sentinel lymph node several hours prior to the operation, and separate perioperative injection of a blue dye. We have developed a combined approach using technetium 99m labeled blue dye (Evans Blue) for use in lymphoscintigraphy that may be injected as a single dose just prior to the operation. METHODS: In an anesthetized rabbit model we dissected a hind limb to display the popliteal node and afferent lymphatic. Technetium 99m Evans Blue ((99m)Tc-EB) (22 MBq; 0.5 mL) was injected subdermally in the dorsum of the paw. Simultaneous digital and gamma camera images were obtained at 14 time intervals to 30 minutes post injection. For each of these time intervals the percentage of radioactivity and percentage blueness of the popliteal node were determined. Urine and afferent lymphatic fluid were analyzed by chromatography. The popliteal node was excised post mortem, placed into solvent solutions and analyzed for blueness and radioactivity. RESULTS: Time-activity curves for radioactivity and time-blueness curves for Evans Blue uptake showed strong correlation (r = 0.958). Lymph analysis suggested (99m)Tc-EB is mainly bound to endogenous proteins. Urine was radioactive but not colored, (99m)Tc-EB being metabolized and excreted in the urine as 1,7-diamino-8-naphthol-2,4-disulfonic acid. Prolonged exposure of node to solvents did not dissociate any blue coloration or radioactivity. CONCLUSIONS: (99m)Tc-EB and Evans Blue are simultaneously retained and concentrated in the sentinel lymph node. This process is rapid and reproducible. (99m)Tc-EB migrates at the same rate as Evans Blue in lymph, where it is transported as bound to endogenous proteins. These dye molecules are metabolized by reductive cleavage in the liver and then excreted renally as colorless, radioactive metabolites. This novel agent has the potential to facilitate lymphatic mapping and subsequent sentinel node biopsy for a range of solid malignancies including breast cancer and melanoma.

Animals↗

PubMed2002

Why certain dyes are useful for localizing the sentinel lymph node.

UNLABELLED: Location of the sentinel lymph node in malignant melanoma and early breast cancer patients requires separate injections of radiocolloid and blue dye. These agents are administered at alternative times because of their different rates of transit. This study has elucidated why particular dyes are absorbed by the lymphatic system from an investigation of dye molecular structure as a function of protein binding ability. METHODS: A dye-protein binding assay was developed using size-exclusion chromatography and ultraviolet spectrophotometry and applied to a series of 20 sulfonic acid group-containing dyes. Radiochemical analyses were also used with 3 99mTc-labeled dyes to rationalize which functional groups are involved in the protein binding reaction. RESULTS: Methylene blue resulted in no protein affinity at 37 degrees C, whereas disulfonate dyes separated by 1 atom such as Patent blue or Indigo carmine gave <30% binding. Optimum protein binding (84%-100%) was achieved with those dyes containing at least 2 sulfonic acid groups separated by 2-6 atoms in their chemical structure. Seven symmetric tetrasulfonic acid azo dyes were examined, including Evans blue, to result in 59%-71% binding. CONCLUSION: Ionizable groups (sulfonic acids) that are present in the structure of dyes are directly involved in dye-protein binding. At the molecular level, there is a sulfonation reaction between sulfonic acid dyes and amino groups on the protein surface to form sulfonamide complexes. This reaction shows how the soluble dyes Evans blue and Patent blue are trapped in lymph after subdermal injection during the sentinel node biopsy procedure.

Animals↗

PubMed2002

Comparison of 99mTc-alafosfalin and 67Ga-citrate in a mouse model of bacterial infection.

BACKGROUND: The antibiotic-peptide (99m)Tc-alafosfalin was assessed as an infection imaging agent in Staphylococcus aureas infected mice by comparison with (99m)Ga-citrate, and also examined for influence of septic status on tracer biodistribution by comparison with normal mice. MATERIAL AND METHODS: Intramuscular doses of S. aureus were administered into the right thigh muscle of mice and the infection was allowed to develop for 20 hours. In separate experiments, (99m)Tc-alafosfalin and (67)Ga-citrate were subsequently administered and allowed to localise. Quantitative organ distribution studies were performed in conjunction with scintigraphic images at 1 and 4 hours post injection. An additional biodistribution with (99m)Tc-alafosfalin in normal mice was also performed. RESULTS: (99m)Tc-alafosfalin was predominantly renal excreted, with low liver, intestine and bone uptake. There was no difference in the uptake of these organs when infected mice were compared with normal mice. (99m)Tc-alafosfalin activity in the intestine at 1 and 4 hours was substantially less than (67)Ga-citrate. For (99m)Tc-alafosfalin, infected/non-infected thigh ratios of 2.8/1.0 and 3.6/1.0 were determined at 1 and 4 hours post injection respectively. (67)Ga-citrate gave ratios of 1.6/1.0 and 3.7/1.0 at the corresponding time points. CONCLUSIONS: (99m)Tc-alafosfalin uptake was more rapid than (67)Ga-citrate, yet diffuse at the infectious sites in mice. The small and juvenile mouse model resulted in uptake of the phosphonic acid tracer by active bone growth areas which may be a disadvantage. This (99m)Tc-antibiotic peptide has potential as an infection imaging agent, and will be investigated further in another rodent infection model in the future.

Journal Article↗

PubMedDate not supplied

Understanding the radiolabelling mechanism of 99mTc-antimony sulphide colloid.

The chemistry of antimony trisulphide colloid (ATC) was examined to elucidate the radiolabelling mechanism with 99mTcO4(-). Ion exchange chromatography and atomic absorption spectrophotometry techniques determined ATC to be resistant to hydrolysis in 0.1M hydrochloric acid (HCl) at 25 degrees C or 100 degrees C (>97% recovery, Sb3+ absent). Hydrogen sulphide gas detected did not participate in the mechanism, where antimony trisulphide and 99mTcO4(-) in HCl/100 degrees C yielded 96% 99mTc-product from a K2S-free formulation (versus 98% when K2S was present). 99mTcO4(-) was reduced >90% by DMSA or dithiothreitol under the same conditions, identifying involvement of thiol groups. Infrared analysis of Re-ATC showed S=O bonds, indicating excess thiol groups at the colloid surface were oxidised at the expense of 99mTcO4(-) reduction.

Antimony↗

PubMedDate not supplied

Radiolabelling and biodistribution of 111In-antimony trisulphide colloid.

The aim of this study was to prepare 111In-antimony trisulphide colloid (111In-ATC) and evaluate its chemical properties in comparison with 99mTc-ATC. After reconstitution of an antimony trisulphide cold Kit with 111In-chloride, 111In-ATC was formed with >95% radiochemical purity in the presence of neutralising phosphate buffer. The product was found to be stable for 4.7 days when stored at room temperature, 80% of radioactive particles were < 20 nm in diameter and the mouse biodistribution assay was identical to that of 99mTc-ATC. In conclusion, pharmaceutical-grade 111In-ATC was successfully prepared and assessed to have properties that are suitable for dual-isotope lymphoscintigraphy studies at the clinical level.

Animals↗

PubMedDate not supplied

Scintigraphic imaging of experimental colitis with technetium-99m-infliximab in the rat.

The radiolabeled monoclonal antibody (99m)Tc-infliximab was assessed as an inflammation imaging agent in a rat colitis model in comparison with (99m)Tc-tin colloid-labeled-leucocytes. (99m)Tc-infliximab and (99m)Tc-tin fluoride colloid-labeled-leucocytes were administered to (n>3) rats previously exposed to 2,4,6-trinitrobenzenesulfonic acid by rectal instillation. Whole body scintigraphic images were acquired and physiological organ assays were performed to obtain quantitative data. Histological examination of colon samples was performed to assess the site and severity of the colitis. In the inflamed colon, (99m)Tc-infliximab resulted in inflamed target (T) to control (C) colon tracer uptake ratios of 2.7 +/- 1.0 (n=5) and 2.6 +/- 0.3 (n=5) at 1 and 4 h post tracer injection respectively. (99m)Tc-leucocytes gave higher ratios of 19.5 +/- 9.9 (n=3) and 41.2 +/- 16.1 (n=5) respectively. (99m)Tc-leucocytes gave higher ratios of 19.5 +/- 9.9 and 41.2 +/- 16.1 at the corresponding time points. (99m)Tc-infliximab accumulated at sites of inflammation in this rat model but not due to a specific tumor necrosis factor-alpha binding mechanism. Although the tracer uptake was lower than radioactive leucocytes, this easily prepared (99m)Tc-monoclonal antibody may have some advantages in imaging inflammatory bowel disease in humans based on its biological activity.

Animals↗