Validation of HPLC and TLC analytical methods to determine radiochemical purity of 99mTc-cAbVCAM1-5, a new experimental radiotracer.

Cardiovascular diseases, including fatal myocardial infarctions from atheromatous plaques, are the primary global mortality cause. Detecting stenotic atheromatous plaques is possible through coronary angiography, but vulnerable plaques with eccentric remodeling are undetectable with current diagnostic methods. Addressing this challenge, our group developed a radiopharmaceutical drug targeting vascular cell adhesion molecule 1 (VCAM-1), radiolabeled with technetium-99m. Given the absence of a monograph in the European Pharmacopoeia, and in order to draft the investigational medicinal product documentation, analytical methods had to be validated by high performance liquid chromatography (HPLC) and thin layer chromatography (TLC) to determine the radiochemical purity (RCP) of 99mTc-cAbVCAM1-5. This study therefore presents the results of the validation of analytical methods obtained in this context. The method validation followed the European Association of Nuclear Medicine (EANM) recommendations adapted from ICH Q2(R1), ensuring conformity with specificity, accuracy, repeatability and intermediate precision, linearity, robustness, quantification limit (LoQ), and range criteria. Regarding the results of specificity, both HPLC and TLC methods demonstrated excellent separation of 99mTc-cAbVCAM1-5 from impurities 99mTcO4-. Accuracy results indicated recovery percentages within the range of 99.52-101.40% for the HPLC and 99.51-101.97% for TLC, ensuring reliable measurements for each concentration of 99mTcO4-. Precision of the methods was validated by assessing repeatability and intermediate precision. Linearity was determined over the usual concentrations range and the correlation coefficient was greater than 0.99 for both methods. The limit of quantification was measured by diluting the 99mTcO4- to obtain a signal-to-noise ratio of around 10:1. Under these conditions, we obtained an LOQ of 2.10 MBq/mL for HPLC and 2Mbq/mL for TLC. In conclusion, the analytical methods developed in this study comply with EANM recommendations. This therefore allows us to correctly assess the radiochemical purity of 99mTc-cAbVCAM1-5, a new radiotracer targeting inflammation in vulnerable plaques.

Development and validation of an alternative procedure for quantitative quality control analysis of 99mTc-radiopharmaceuticals using a Geiger Müller counter.

BACKGROUND AND OBJECTIVES: In a hospital radiopharmacy with 2a operational level, including the preparation of radiopharmaceuticals from prepared and approved reagent kits, it is common to have a single activimeter or dose calibrator for labeling and fractionation, and to perform the quality controls of the 99mTc-radiopharmaceuticals. In certain cases, the accumulation of radioactive material or accidental contamination of the work area causes the background to exceed the limits to carry out the radiochemical purity analyses and it is necessary to look for viable alternatives. In this work, a Geiger Müller detector (equipped with a probe for measuring surface contamination) frequently used for radioprotection purposes, was validated as an alternative and its performance was compared against the activimeter for 99mTc-radiopharmaceuticals. MATERIALS AND METHODS: Using [99mTc]pertechnetate, systematic studies of error analyses and detector response to activity concentration, activity and measurement time were carried out in liquid matrices and in paper. The results were compared against an activimeter calibrated for [99mTc]Tc. RESULTS: The developed method was used to determine the radiochemical purity of the compounds [99mTc]Tc-MDP and [99mTc]Tc-MIBI by ascending paper chromatography tests, obtaining comparable values to those measured with an activimeter in the same system (within 1% uncertainty) and using the method of vial partitioning in a dedicated equipment. CONCLUSIONS: This work demonstrates that a Geiger Müller detector with a probe for measuring surface contamination can be adequately used to replace other equipment in the control of radiochemical purity in the hospital radiopharmacy.

The Removal of Pertechnetate from Aqueous Solution by Synthetic Hydroxyapatite: The Role of Reduction Reagents and Organic Ligands.

The use of knowledge from technetium radiochemistry (even from nuclear medicine applications) allows us to select an sorbent for 99mTc radionuclide sorption, which is hydroxyapatite. Using radioisotope indication, the 99mTcO4- sorption process on synthetic hydroxyapatite was studied by the batch method in the presence of SnCl2 and FeSO4 reducing agents. The complexing organic ligands' effect on the 99mTcO4- sorption under reducing conditions was investigated. In the presence of Sn2+ ions without the addition of organic ligand, the sorption percentage reached above 90% independently of the environment. In the presence of Fe2+ ions without the addition of organic ligand, the sorption of 99mTcO4- was significantly lower and was at approximately 6%, depending on the concentration of Fe2+ ions in solution. The effect of complexing organic ligands on the 99mTcO4- sorption on hydroxyapatite from the aqueous solution, acetate buffer and phosphate buffer decreases in the following order for Sn2+: oxalic acid > ethylenediaminetetraacetic acid > ascorbic acid. In the presence of Fe2+ ions without organic ligands, the sorption reached up to 15% depending on the composition of the solution. The addition of oxalic acid and ascorbic acid increased the sorption up to 80%. The ethylenediaminetetraacetic acid had no significant effect on the sorption of technetium on hydroxyapatite.

Transferability of a two-strip method for the quality control of technetium-99m mercaptoacetyltriglycine ([99mTc]Tc-MAG3).

OBJECTIVES: Technetium-99m mercapto-acetyl-triglycine ([99mTc]Tc-MAG3) is a radiopharmaceutical diagnostic agent used in nuclear medicine intended for the exploration of nephrological and urological disorders. Patient safety and reliability of this imaging procedure especially depend on the radiochemical purity (RCP) of the [99mTc]Tc-MAG3 preparation. Recently, the Summary of Product Characteristics (SPC) of NephroMAG, a kit dedicated to [99mTc]Tc-MAG3 preparation, proposed a two-strip thin layer chromatography (TLC) based quality control (QC) method. Also, Straub et al recently proposed another TLC based QC method. We sought to evaluate the transferability of these QC methods in our hospital radiopharmacy and compared them to our currently employed TLC method and radio-HPLC (high-pressure liquid chromatography) to select the most appropriate in the context of hospital radiopharmacy. METHODS: Ten consecutive [99mTc]Tc-MAG3 preparations were controlled using: HPLC combined with a radiodetector (radio-HPLC), a single strip TLC method (method 1) in current use in our centre, a two-strip TLC method described in the SPC (method SPC) and a two-strip TLC method (method 2) described by Straub et al. Quantitative results for the four tested QC methods were measured and compared in terms of RCP (%), sodium pertechnetate ([99mTc]TcO4 -) (%) and duration of analysis (min). RESULTS: RCP was significantly different between method SPC and radio-HPLC (p<0.001) and method 2 (p<0.001). Also, the percentage of [99mTc]TcO4 - was statistically different between the radio-HPLC and the method SPC (p<0.001), but not with the method 1 and method 2 group (p>0.05). The duration of analysis (min) was significantly different between the four QC procedures (p<0.001) with method 2 and method SPC being the quickest. CONCLUSIONS: Our study showed it is possible to transfer and select a quick and reliable QC method for the preparation of NephroMAG kits in our centre. We therefore advise the widespread use of the method from Straub et al in hospital radiopharmacies.

99mTc internal contaminations measurements among nuclear medicine medical personnel during ventilation - perfusion SPECT lung scans.

This paper presents results of measurements of 99mTc activity concentration in air and nuclear medical personnel blood during ventilation-perfusion SPECT lung scans. 99mTc activity measurements were conducted at the Nuclear Medicine Department, John Paul II Hospital, Krakow. Technicians and nurses who perform examinations were equipped with personal aspirators enabling air sampling to determine the radiation exposure at their workplaces. Measurements allowed to evaluate the concentration of 99mTc in 14 air samples and it ranged from 7800 ± 600 to 10,000 ± 1000 Bq m-3 for air samples collected by technicians and from 390 ± 30 to 600 ± 40 Bq m-3 for air samples collected by nurses. In addition 99mTc concentrations in blood of medical personnel were determined in 24 samples. For technicians the maximum 99mTc blood concentration levels reached 920 ± 70 Bq L-1 and 1300 ± 100 Bq L-1. In the case of nurses, the maximum estimated activity concentrations were about ten times lower, namely 71 ± 7 Bq L-1 and 39 ± 3 Bq L-1. Although the intakes appear to be relatively high, the resulting annual effective doses are about 34 µSv for technicians and only 2 µSv for nurses.

Sentinel lymph node detection by means of indocyanine green using the Karl Storz VITOM® fluorescence camera: a comparison between primary sentinel lymph node biopsy versus sentinel lymph node biopsy after neoadjuvant chemotherapy.

PURPOSE: The usage of radioactive Technetium99m (Tc99m) colloid for the purpose of sentinel lymph node biopsy (SLNB) in early breast cancer is considered the gold standard in Germany. However, new tracers, such as near-infrared (NIR) imaging agents like indocyanine green (ICG) could offer an alternative in future, as they overcome drawbacks associated with radioactive Technetium99m (Tc99m) like limited availability, high costs and radioactivity exposure for both patients and surgeons. METHODS: In this double-arm retrospective study, we sought to establish the usefulness of indocyanine green as an alternative or an addition to the conventional Technetium99m (Tc99m) in the identification of the SLN in early breast cancer. RESULTS: Among the 161 patients who underwent primary SLNB, 34 patients had at least 1 SLN with metastasis. Among these patients with SLN metastasis, 33 had the SLN detected by ICG; while 31 had the SLN detected by Tc99m. The conventional Technetium99m radiotracer failed to detect 2 patients with metastasis in this Arm of the study. Among the 87 patients who underwent SLNB after NACT, 13 patients had at least 1 SLN with metastasis. Among these 13 patients with SLN metastasis, ICG and Tc99m had detected the SLN among 12 patients, while 1 patient had been detected by ICG alone. CONCLUSIONS: Our results show that ICG is as effective as the radioisotope for SLNB even among patients who have undergone NACT. This trial is registered with the German Clinical Trial Register, ID: DRKS00013606.

Extremity Exposure with 99mTc - Labelled Radiopharmaceuticals in Diagnostic Nuclear Medicine.

BACKGROUND: Extremity exposures may raise the risk of cancer induction among radiographers involved in the preparation and administration of technetium-99m labelled radiopharmaceuticals. OBJECTIVE: To estimate finger doses on radiographers at a South African tertiary hospital. METHODS: Adhesive tape was used to securely fix a calibrated thermoluminescent dosimeter (TLD) on fingertips and bases of ring and index fingers of both hands of five radiographers who prepared and administered technetium-99m labelled radiopharmaceuticals. Rubber gloves were worn to avoid TLD contamination. TLDs doses were read with a Harsaw TLD Reader (Model 3500) after a week. RESULTS: Five radiographers prepared and administered technitium-99m labelled radiopharmaceuticals (activity range; 78.20 GBq - 132.78 GBq during a one-week measurement period). A radiographer handling 132.78 GBq received 4.74±0.52 mSv on both hands; 5.52, 4.55, 5.11 and 4.60 mSv on the fingertip of the index finger of the dominant hand (FIDH), fingertip of the ring finger of the dominant hand (FRDH), fingertip of the index finger of the non-dominant hand (FINDH) and fingertip of the ring finger of the non-dominant hand (FRNDH), respectively. The respective doses received on the finger bases were 4.50 mSv, 4.60, 4.21 and 3.48 mSv. The radiographer handling 78.20 GBq received 0.85±0.18 mSv on both hands, 1.04, 1.17, 0.77 and 1 mSv for the FIDH, FRDH, FINDH and FRNDH, respectively, while respective doses for the bases were 0.8, 0.9, 0.6 and 0.8 mSv. CONCLUSION: The extremity exposures were below the annual limit (500 mSv). However, the use of syringe shields could still reduce the finger doses further.

Phase I Study of 99mTc-ADAPT6, a Scaffold Protein-Based Probe for Visualization of HER2 Expression in Breast Cancer.

Radionuclide molecular imaging of human epidermal growth factor receptor type 2 (HER2) expression may help to stratify breast and gastroesophageal cancer patients for HER2-targeting therapies. Albumin-binding domain-derived affinity proteins (ADAPTs) are a new type of small (46-59 amino acids) protein useful as probes for molecular imaging. The aim of this first-in-humans study was to evaluate the biodistribution, dosimetry, and safety of the HER2-specific 99mTc-ADAPT6. Methods: Twenty-nine patients with primary breast cancer were included. In 22 patients with HER2-positive (n = 11) or HER2-negative (n = 11) histopathology, an intravenous injection of 385 ± 125 MBq of 99mTc-ADAPT6 was performed, randomized to an injected protein mass of either 500 µg (n = 11) or 1,000 µg (n = 11). Planar scintigraphy followed by SPECT imaging was performed after 2, 4, 6, and 24 h. An additional cohort (n = 7) was injected with 165 ± 29 MBq (injected protein mass, 250 µg), and imaging was performed after 2 h only. Results: Injections of 99mTc-ADAPT6 were well tolerated at all mass levels and not associated with adverse effects. 99mTc-ADAPT6 cleared rapidly from the blood and most other tissues. The normal organs with the highest accumulation were the kidney, liver, and lung. Effective doses were 0.009 ± 0.002 and 0.010 ± 0.003 mSv/MBq for injected protein masses of 500 and 1,000 µg, respectively. Injection of 500 µg resulted in excellent discrimination between HER2-positive and HER2-negative tumors as early as 2 h after injection (tumor-to-contralateral breast ratio, 37 ± 19 vs. 5 ± 2; P < 0.01). The tumor-to-contralateral breast ratios for HER2-positive tumors were significantly (P < 0.05) higher for an injected mass of 500 µg than for either 250 or 1,000 µg. Conclusion: Injections of 99mTc-ADAPT6 are safe and associated with low absorbed and effective doses. A protein dose of 500 µg is preferable for discrimination between tumors with high and low expression of HER2. Further studies are justified to evaluate whether 99mTc-ADAPT6 can be used as an imaging probe to stratify patients for HER2-targeting therapy in areas where PET imaging is not readily available.

Evaluation of Radiation dosimetry of 99mTc-HYNIC-PSMA and imaging in prostate cancer.

This study aims to evaluate the radiation dosimetry of a new technetium-99m‒labelled small-molecule inhibitor of prostate-specific membrane antigen (HYNIC-Glu-Urea-A, 99mTc-HYNIC-PSMA) and its feasibility as a tumor-imaging agent in prostate cancer (PCa) patients. A total of 15 PCa patients were enrolled in this study. For the dosimetry study, 5 PCa patients received whole-body planar scans at 0.5 h, 1 h, 2 h, 4 h and 8 h after 99mTc-HYNIC-PSMA injection. The Dosimetry Toolkit (GE, Milwaukee) was used to process the data and segment the organs in the SPECT/CT images, which were then projected onto planar images. The organ-specific absorbed doses, total-body absorbed doses and 99mTc-HYNIC-PSMA effective doses of patients were calculated using OLINDA/EXM 1.1 software. Whole-body SPECT/CT images were also acquired from additional 10 prostate patients to investigate the feasibility of 99mTc-HYNIC-PSMA for imaging tumors by calculating the ratio of tumor-to-background tracer uptake at 2 h after 740 MBq administration. The total-body absorbed dose was 1.54E-03 ± 2.43E-04 mGy/MBq, and the effective dose was 3.72E-03 ± 4.5E-04 mSv/MBq. Compared to published studies of other similar PSMA tracers and 99mTc-targeted conventional tracers, the absorbed doses of 99mTc-HYNIC-PSMA in all organs showed that it could be used safely in the human body. In addition, 99mTc-HYNIC-PSMA showed high tracer uptake (with a tumor-to-background ratio of 9.42 ± 2.62) in the malignant lesions of PCa patients, making it a promising radiopharmaceutical imaging method for site-specific management of PCa.

Preparation, radiolabeling with 99mTc and 67Ga and biodistribution studies of albumin nanoparticles covered with polymers. / Preparación, radiomarcaje con 99mTc y 67Ga y estudios de biodistribución de nanopartículas de albúmina con recubrimientos poliméricos.

OBJECTIVE: To optimize radiolabeling with 99mTc and 67Ga of albumin nanoparticles coated with 4 differents synthetic polymers and to evaluate their stability in vivo and in vitro, as well as their biodistribution in vivo after intravenous administration. MATERIAL AND METHODS: The nanoparticles were prepared using albumin and NOTA-modified albumin by the desolvation method and coated with 4 different polymers; HPMC, GMN2, GPM2 and GTM2. They were purified, lyophilized and characterized. Radiolabelling with 99mTc was perfomed with 74 MBq of 99mTc sodium pertechnetate, previously reduced with and acid solution of tin chloride at different concentrations (0.003, 0.005, 0.007, 0.01, 0.05 and 0.1mg/ml) and at different times (5, 10, 15, 30 and 60minutes) and temperatures (room temperature, 40°C and 60°C). Radiolabelling with 67Ga was perfomed by incubation of the nanoparticles with 37 MBq of 67Gallium chloride (obtained from commercial gallium-67 citrate) at different times (10 and 30minutes) and temperatures (room temperature, 30°C and 60°C), and posterior purification with microconcentrators. The radiochemical purity was evaluated by TLC. Stability studies of radiolabeled nanoparticles in physiological serum and blood plasma were perfomed. Biodistribution studies of nanoparticles coated with GPM2 polymer were carried out in Wistar rats after intravenous administration of the nanoparticles. Control animals were carried out with 99mTc sodium pertechnetate and 67Ga chloride. To do so, the animals were killed and activity in organs was measured in a gamma counter. RESULTS: 99mTc labeling was carried out optimally with a tin concentration of 0.007mg/ ml for the GPM2 nanoparticles and 0.005mg / ml for the rest of the formulations, with a radiolabelling time of 10minutes at room temperature. In the case of 67Ga the label was optimized at 30° C temperature and 30minutes of incubation. In both cases the radiochemical purity obtained was greater than 97%. The nanoparticles showed high stability in vitro after 48hours of labeling (70% nanoparticles labeled with 99mTc and 90% those labeled with 67Ga). Biodistribution studies of nanoparticles 99mTc -GPM2 and 67Ga -NOTA-GPM2 showed a high accumulation of activity in the liver at 2 and 24hours after intravenous administration. CONCLUSION: The labeling procedure with 99mTc and 67Ga of albumin and albumin modified with NOTA nanoparticles allows obtaining nanoparticles with high labeling yields and adequate in vitro stability, allowing their use for in vivo studies.

Active Anion Delivery by Self-Propelled Microswimmers.

Self-propelled micro- and nanomachines are at the forefront of materials research, branching into applications in biomedical science and environmental remediation. Cationic frameworks enabling the collection and delivery of anionic species (A-) are highly required, due to the large variety of life-threatening pollutants, such as radioactive technetium and carcinogenic chromium, and medicines, such as dexamethasone derivatives with negative charges. However, such autonomous moving carriers for active transport of the anions have been barely discussed. A polymeric viologen (PV++)-consisting of electroactive bicationic subunits-is utilized in a tubular autonomous microswimmer to selectively deliver A- of different sizes and charge densities. The cargo loading is based on a facile anion exchange mechanism. The packed crystal structure of PV++ allows removal of an exceptionally high quantity of anions per one microswimmer (2.55 × 10-13 mol anions per microswimmer), a critical factor often neglected regarding the real-world application of microswimmers. Notably, there was virtually no leakage of anions during the delivery process or upon keeping the loaded microswimmers under ambient conditions for at least 4 months. Multiple release mechanisms, compatible with different environments, including electrochemical, photochemical, and a metathesis reaction, with high efficiencies up to 98% are introduced. Such functional autonomous micromachines provide great promise for the next generation of functional materials for biomedical and environmental applications.

Using porous iron composite (PIC) material to immobilize rhenium as an analogue for technetium.

Technetium (99Tc), a uranium-235 (235U) and plutonium-239 (239Pu) fission product, is a primary risk driver in low level radioactive liquid waste at U.S. Department of Energy sites. Previous studies have shown success in using Zero Valent Iron (ZVI) to chemically reduce and immobilize redox sensitive groundwater contaminants. Batch and column experiments were performed to assess the ability of a novel porous iron composite material (PIC) to immobilize Tc(VII) in comparison with two commercial Fe oxide sorbents and reagent grade ZVI in the presence and absence of NO3-, a competing oxidized species that is often found in high concentrations in liquid nuclear waste. Perrhenate (ReO4-) was used as a non-radioactive chemical analogue for pertechnetate (TcO4-) under both oxic and anoxic test conditions. The PIC powder was the most effective at immobilizing Re(VII) under all batch test conditions. The presence of nitrate (NO3-) slowed the removal of ReO4- from solution, presumably through chemical reduction and precipitation. Even so, the PIC and ZVI were effective at removing both Re(VII) and NO3- completely from solution. Nitrate was reduced to NH3 with very little nitrite (NO2-) buildup during equilibration. Significant Re immobilization was observed in the column tests containing PIC sorbent, even though inlet solutions were in equilibrium with O2. The presence of NO3- hastened Re breakthrough, while NO3- reduction to NH3 was observed. The results suggest that PIC and ZVI would be the most effective at the removal of TcO4- from contaminated groundwater sites.

99mTc activity concentrations in room air and resulting internal contamination of medical personnel during ventilation-perfusion lung scans.

This paper presents the results of measurements of 99mTc activity concentrations in indoor air in a nuclear medicine department and resulting estimated 99mTc intake by medical personnel. 99mTc air activity measurements were conducted at the Nuclear Medicine Department, John Paul II Hospital, Krakow, Poland, during ventilation-perfusion SPECT lung scans. Technetium from the air was collected by means of a mobile aerosol sampler with a Petryanov filter operating at an average flow rate of 10 dm3 min-1. Measured activities ranged from 99 ± 11 to 6.1 ± 0.5 kBq m-3. The resulting daily average intake of 99mTc by medical staff was estimated to be 5.4 kBq, 4.4 kBq, 3.0 kBq and 2.5 kBq, respectively, for male technicians, female technicians, male nurses and female nurses. Corresponding annual effective doses were 1.6 µSv for technicians and 1 µSv for nurses. The highest equivalent dose values were determined for extrathoracic (ET) airways: 5 µSv and 10 µSv for nurses and technicians, respectively. It is concluded that estimated annual absorbed doses are over three orders of magnitude lower than the dose limit established in the Polish Atomic Law Act and in recommendations of the International Commission on Radiological Protection for medical staff.

Ultratrace Determination of 99Tc in Small Natural Water Samples by Accelerator Mass Spectrometry with the Gas-Filled Analyzing Magnet System.

In the frame of studies on the safe disposal of nuclear waste, there is a great interest for understanding the migration behavior of 99Tc. 99Tc originating from nuclear energy production and global fallout shows environmental levels down to 107 atoms/g of soil (∼2 fg/g). Extremely low concentrations are also expected in groundwater after diffusion of 99Tc through the bentonite constituting the technical barrier for nuclear waste disposal. The main limitation to the sensitivity of the mass spectrometric analysis of 99Tc is the background of its stable isobar 99Ru. For ultratrace analysis, the Accelerator Mass Spectrometry (AMS) setup of the Technical University of Munich using a Gas-Filled Analyzing Magnet System (GAMS) and a 14 MV Tandem accelerator is greatly effective in suppressing this interference. In the present study, the GAMS setup is used for the analysis of 99Tc in samples of the seawater reference material IAEA-443, a peat bog lake, and groundwater from an experiment of in situ diffusion through bentonite in the controlled zone of the Grimsel Test Site (GTS) within the Colloid Formation and Migration (CFM) project. With an adapted chemical preparation procedure, measurements of 99Tc concentrations at the fg/g levels with a sensitivity down to 0.5 fg are accomplished in notably small natural water samples. The access to these low concentration levels allows for the long-term monitoring of in situ tracer tests over several years and for the determination of environmental levels of 99Tc in small samples.

Sorption of 137Cs, 90Sr, Se, 99Tc, 152(154)Eu, 239(240)Pu on fractured rocks of the Yeniseysky site (Nizhne-Kansky massif, Krasnoyarsk region, Russia).

The study demonstrates the effect of sorption properties of fractured host rocks from the Yeniseysky site (Nizhne-Kansky rock massif, Krasnoyarsk region) on the migration of dissolved radioactive components (137Cs, 90Sr, 79Se, 99Tc, 152(154)Eu, 239(240)Pu) in the deep geological conditions of a high-level radioactive waste repository. Estimates of radionuclide distribution coefficients between the aqueous solution and fractured rocks obtained from sorption experiments were used to study the influence of various petrographic types and fracture-filling substances on the retardation of radioactive components by different fractured rock units of Nizhne-Kansky rock massif. Based on the results of sorption experiments, we concluded that the type and attributes of rock discontinuities, as well as the mineral composition of the material in fractures, are crucial for the immobilization of radionuclides during their migration through a geological environment.

Diagnosis of LVAD Thrombus using a High-Avidity Fibrin-Specific 99mTc Probe.

Treatment of advanced heart failure with implantable LVADs is increasing, driven by profound unmet patient need despite potential serious complications: bleeding, infection, and thrombus. The experimental objective was to develop a sensitive imaging approach to assess early thrombus accumulation in LVADs under operational high flow and high shear rates. Methods: A monomeric bifunctional ligand with a fibrin-specific peptide, a short spacer, and 99mTc chelating amino acid sequence (F1A) was developed and compared to its tetrameric PEG analogue (F4A). Results:99mTc attenuation by LVAD titanium (1 mm) was 23%. 99mTc-F1A affinity to fibrin was Kd ~10 µM, whereas, the bound 99mTc-F4A probe was not displaced by F1A (120,000:1). Human plasma interfered with 99mTc-F1A binding to fibrin clot (p<0.05) in vitro, whereas, 99mTc-F4A targeting was unaffected. The pharmacokinetic half-life of 99mTc-F4A was 28% faster (124±41 min) than 99mTc-F1A (176±26 min) with both being bioeliminated through the urinary system with negligible liver or spleen biodistribution. In mice with carotid thrombus, 99mTc-F4A binding to the injured carotid was much greater (16.3±3.3 %ID/g, p=0.01) than that measured with an irrelevant negative control, 99mTc-I4A (3.4±1.6 %ID/g). In an LVAD mock flow-loop (1:1, PBS:human plasma:heparin) operating at maximal flow rate, 99mTc-F4A bound well to phantom clots in 2 min (p<0.05), whereas 99mTc-F1A had negligible targeting. Excised LVADs from patients undergoing pump exchange or heart transplant were rewired, studied in the mock flow loop, and found to have spatially variable fibrin accumulations in the inlet and outlet cannulas and bearings. Conclusions:99mTc-F4A is a high-avidity prototype probe for characterizing thrombus in LVADs that is anticipated to help optimize anticoagulation, reduce thromboembolic events, and minimize pump exchange.

Decontamination of rat and human skin experimentally contaminated with 99mTc, 201Tl and 131I radionuclides using "Dermadecon" - a skin decontamination kit: an efficacy study.

Radioactive skin contamination is one of the most likely risks which occurs after accidental or occupational radiological accidents apart from internal contamination. In such cases where the radioactive contamination has occurred, the person who is contaminated should be decontaminated as early as possible to reduce the damaging health effects of radiation. In the present study, the decontamination efficiency of a developed skin decontamination kit "dermadecon" has been evaluated in animal models and human subjects using gamma scintigraphy. Decontamination efficiency (percentage of the radioactive contaminant removed) was calculated for each radioactive isotope of the study and compared with control where general washing procedure was followed using liquid and soap. The effectiveness of the kit was calculated in animal model with respect to 99mTc-sodium-pertechnetate (99mTcO4-), 201TlCl and 131I and was found 92.84 ± 4.9%, 91.18 ± 3.23% and 94.67 ± 2.92%, respectively. Whereas, in case of human skin, the decontamination efficiency for 99mTcO4- was observed to be 95.00 ± 3.21%. On the basis of findings from the study, it can be concluded that the decontamination agents of the used skin decontamination kit are effective for removal of localized radioactive contaminants from skin, as compared with normal decontamination using soap and water.

A new method for studying cryptococcosis in a murine model using 99mTc-Cryptococcus gattii.

Cryptococcus gattii is one of the etiologic agents of cryptococcosis, a systemic mycosis that occurs in healthy and immunosuppressed humans and animals worldwide. Primary pulmonary infection caused by C. gattii is usually followed by fungal dissemination to the central nervous system, resulting in high mortality rates. In this context, animal models of cryptococcosis are useful in the study of fungal pathogenesis and host response against the pathogen, and for testing novel therapeutic options. The most frequently applied method to study fungal dissemination from the lungs to other organs is by culturing tissues, which is not accurate for the detection and quantification of fungal load at early stages of the infection. To overcome this problem, the purpose of this study was to develop a new method for the quantification of Cryptococcus dissemination. One C. gattii strain was efficiently radiolabeled with technetium-99m (99mTc), without affecting viability of the cells. Further, the 99mTc-C. gattii (111 MBq) strain was used to infect mice by intratracheal and intravenous route for biodistribution studies. 99mTc-C. gattii was successfully used in detection of the yeast in the brain of mice 6 hours postinoculation, while the detection using colony forming units was possible only 24 hours postinfection. Our results provided an alternative method that could be applied in further investigations regarding the efficacy of antifungals, fungal virulence, and host-pathogen interactions.

Effects of hydrated lime on radionuclides stabilization of Hanford tank residual waste.

Chemical stabilization of tank residual waste is part of a Hanford Site tank closure strategy to reduce overall risk levels to human health and the environment. In this study, a set of column leaching experiments using tank C-104 residual waste were conducted to evaluate the leachability of uranium (U) and technetium (Tc) where grout and hydrated lime were applied as chemical stabilizing agents. The experiments were designed to simulate future scenarios where meteoric water infiltrates through the vadose zones into the interior of the tank filled with layers of grout or hydrated lime, and then contacts the residual waste. Effluent concentrations of U and Tc were monitored and compared among three different packing columns (waste only, waste + grout, and waste + grout + hydrated lime). Geochemical modeling of the effluent compositions was conducted to determine saturation indices of uranium solid phases that could control the solubility of uranium. The results indicate that addition of hydrated lime strongly stabilized the uranium through transforming uranium to a highly insoluble calcium uranate (CaUO4) or similar phase, whereas no significant stabilization effect of grout or hydrated lime was observed on Tc leachability. The result implies that hydrated lime could be a great candidate for stabilizing Hanford tank residual wastes where uranium is one of the main concerns.