Dynamic Effects of Tank Waste Aging on Radionuclide-Complexant interactions

The overall objective of this project is to provide a scientific basis for safely processing complexant-containing high-level tank wastes for disposal. Our key goals are to identify methods for preparing realistic complexant-containing tank waste simulants, and to use those simulants to determine the relative importance of organic complexants and their breakdown products on the partitioning of important radionuclides. Complexant containing waste simulants are aged by various chemical processes and compared to samples of actual Hanford tank waste. This work will help to identify important aging processes in the tanks and provide the basis for developing successful treatment strategies

Dynamic Effects of Tank Waste Aging on Radionuclide-Complexant Interactions - Final Report - 10/01/1997 - 10/01/2000

The long-range objective of this project is to provide a scientific basis for safely processing high-level nuclear tanks wastes for disposal. Our goals are to identify a means to prepare realistic simulant formulations for complexant-containing Hanford tank wastes, and then use those simulants to determine the relative importance of various organic complexants and their breakdown products on the partitioning of important radionuclides. The harsh chemical and radiolytic environment in high-level waste tanks alters both the organic complexants and the metal species, producing radionuclide-chelator complexes that resist standard separation methods. A detailed understanding of the complexation reactions of the key radionuclides in tank wastes would allow for reliable, science-based solutions for high-level waste processing, but a key problem is that tank waste samples are exceedingly difficult to obtain, transport and handle in the laboratory. In contrast, freshly-prepared simulated wastes are safe and readily obtained, but they do not reproduce the partitioning behavior of actual tank waste samples. For this project, we will first artificially age complexant-containing tank waste simulants using microwave, ultrasound, and photolysis techniques that can be applied in any standard laboratory. The aged samples will be compared to samples of actual Hanford tank wastes to determine the most realistic aging method, on the basis of the organic fragments present, and the oxidation states and partitioning behavior of important radionuclides such as 90Sr, 99Tc, and 239Pu. Our successful completion of this goal will make it possible for scientists in academic and industrial laboratories to address tank waste remediation problems without the enormous costs and hazards associated with handling actual tank waste samples. Later, we will use our simulant aging process to investigate the relative effects of chelator degradation products on the partitioning of important radionuclides from the waste. Using NMR-active labels in the chelators, we will use a combinatorial approach of generating multiple chelator fragments in a single experiment and then determining which, if any, of the fragments have a negative effect on the separations chemistry. Our successful completion of this goal will specifically identify the most problematic organic fragments in complexant-containing waste and provide the basis for developing successful treatment strategies for these wastes

Ribavirin Reveals a Lethal Threshold of Allowable Mutation Frequency for Hantaan Virus▿

The broad spectrum of antiviral activity of ribavirin (RBV) lies in its ability to inhibit IMP dehydrogenase, which lowers cellular GTP. However, RBV can act as a potent mutagen for some RNA viruses. Previously we have shown a lack of correlation between antiviral activity and GTP repression for Hantaan virus (HTNV) and evidence for RBV's ability to promote error-prone replication. To further explore the mechanism of RBV, GTP levels, specific infectivity, and/or mutation frequency was measured in the presence of RBV, mycophenolic acid (MPA), selenazofurin, or tiazofurin. While all four drugs resulted in a decrease in the GTP levels and infectious virus, only RBV increased the mutation frequency of viral RNA (vRNA). MPA, however, could enhance RBV's mutagenic effect, which suggests distinct mechanisms of action for each. Therefore, a simple drop in GTP levels does not drive the observed error-prone replication. To further explore RBV's mechanism of action, we made a comprehensive analysis of the mutation frequency over several RBV concentrations. Of importance, we observed that the viral population reached a threshold after which mutation frequency did not correlate with a dose-dependent decrease in the level of vRNA, PFU, or [RTP]/[GTP] (where RTP is ribavirin-5′-triphosphate) over these same concentrations of RBV. Modeling of the relationship of mutation frequency and drug concentration showed an asymptotic relationship at this point. After this threshold, approximately 57% of the viral cDNA population was identical to the wild type. These studies revealed a lethal threshold, after which we did not observe a complete loss of the quasispecies structure of the wild-type genome, although we observed extinction of HTNV

Preclinical Metabolism, Pharmacokinetics and In Vivo Analysis of New Blood-Brain-Barrier Penetrant Fingolimod Analogues: FTY720-C2 and FTY720-Mitoxy.

Parkinson's disease (PD) is a neurodegenerative aging disorder in which postmortem PD brain exhibits neuroinflammation, as well as synucleinopathy-associated protein phosphatase 2A (PP2A) enzymatic activity loss. Based on our translational research, we began evaluating the PD-repurposing-potential of an anti-inflammatory, neuroprotective, and PP2A stimulatory oral drug that is FDA-approved for multiple sclerosis, FTY720 (fingolimod, Gilenya®). We also designed two new FTY720 analogues, FTY720-C2 and FTY720-Mitoxy, with modifications that affect drug potency and mitochondrial localization, respectively. Herein, we describe the metabolic stability and metabolic profiling of FTY720-C2 and FTY720-Mitoxy in liver microsomes and hepatocytes. Using mouse, rat, dog, monkey, and human liver microsomes the intrinsic clearance of FTY720-C2 was 22.5, 79.5, 6.0, 20.2 and 18.3 μL/min/mg; and for FTY720-Mitoxy was 1.8, 7.8, 1.4, 135.0 and 17.5 μL/min/mg, respectively. In hepatocytes, both FTY720-C2 and FTY720-Mitoxy were metabolized from the octyl side chain, generating a series of carboxylic acids similar to the parent FTY720, but without phosphorylated metabolites. To assess absorption and distribution, we gave equivalent single intravenous (IV) or oral doses of FTY720-C2 or FTY720-Mitoxy to C57BL/6 mice, with two mice per time point evaluated. After IV delivery, both FTY720-C2 and FTY720-Mitoxy were rapidly detected in plasma and brain; and reached peak concentrations at the first sampling time points. After oral dosing, FTY720-C2 was present in plasma and brain, although FTY720-Mitoxy was not orally bioavailable. Brain-to-plasma ratio of both compounds increased time-dependently, suggesting a preferential partitioning to the brain. PP2A activity in mouse adrenal gland increased ~2-fold after FTY720-C2 or FTY720-Mitoxy, as compared to untreated controls. In summary, FTY720-C2 and FTY720-Mitoxy both (i) crossed the blood-brain-barrier; (ii) produced metabolites similar to FTY720, except without phosphorylated species that cause S1P1-mediated-immunosuppression; and (iii) stimulated in vivo PP2A activity, all of which encourage additional preclinical assessment