Cellular pharmacology of multi- and duplex drugsconsisting of ethynylcytidine and 5-fluoro-2′-deoxyuridine

Prodrugs can have the advantage over parent drugs in increased activation and cellular uptake. The multidrug ETC-L-FdUrd and the duplex drug ETC-FdUrd are composed of two different monophosphate-nucleosides, 5-fluoro-2′deoxyuridine (FdUrd) and ethynylcytidine (ETC), coupled via a glycerolipid or phosphodiester, respectively. The aim of the study was to determine cytotoxicity levels and mode of drug cleavage. Moreover, we determined whether a liposomal formulation of ETC-L-FdUrd would improve cytotoxic activity and/or cleavage. Drug effects/cleavage were studied with standard radioactivity assays, HPLC and LC-MS/MS in FM3A/0 mammary cancer cells and their FdUrd resistant variants FM3A/TK−. ETC-FdUrd was active (IC50 of 2.2 and 79 nM) in FM3A/0 and TK− cells, respectively. ETC-L-FdUrd was less active (IC50: 7 nM in FM3A/0 vs 4500 nM in FM3A/TK−). Although the liposomal formulation was less active than ETC-L-FdUrd in FM3A/0 cells (IC50:19.3 nM), resistance due to thymidine kinase (TK) deficiency was greatly reduced. The prodrugs inhibited thymidylate synthase (TS) in FM3A/0 cells (80–90%), but to a lower extent in FM3A/TK− (10–50%). FdUMP was hardly detected in FM3A/TK− cells. Inhibition of the transporters and nucleotidases/phosphatases resulted in a reduction of cytotoxicity of ETC-FdUrd, indicating that this drug was cleaved outside the cells to the monophosphates, which was verified by the presence of FdUrd and ETC in the medium. ETC-L-FdUrd and the liposomal formulation were neither affected by transporter nor nucleotidase/phosphatase inhibition, indicating circumvention of active transporters. In vivo, ETC-FdUrd and ETC-L-FdURd were orally active. ETC nucleotides accumulated in both tumor and liver tissues. These formulations seem to be effective when a lipophilic linker is used combined with a liposomal formulation

Search for invisible modes of nucleon decay in water with the SNO+ detector

This paper reports results from a search for nucleon decay through invisible modes, where no visible energy is directly deposited during the decay itself, during the initial water phase of SNO+. However, such decays within the oxygen nucleus would produce an excited daughter that would subsequently deexcite, often emitting detectable gamma rays. A search for such gamma rays yields limits of 2.5×1029  y at 90% Bayesian credibility level (with a prior uniform in rate) for the partial lifetime of the neutron, and 3.6×1029  y for the partial lifetime of the proton, the latter a 70% improvement on the previous limit from SNO. We also present partial lifetime limits for invisible dinucleon modes of 1.3×1028  y for nn, 2.6×1028  y for pn and 4.7×1028  y for pp, an improvement over existing limits by close to 3 orders of magnitude for the latter two

Decomposition of roots and shoots of perennial grasses and annual barley-separately or in two residue mixes

Little is known about the decomposition rates of shoot and root residues of perennial grasses. This knowledge is important to estimate the carbon sequestration potential of the grasses. An incubation experiment was carried out in a sandy clay loam with shoot and root residues of three native perennial grasses (Wallaby grass, Stipa sp. and Kangaroo grass) and the annual grass barley either separately or in mixtures of two residues. Respiration rate was measured over 18 days, and microbial C and available N were measured on days 0 and 18. Decomposition was lower for roots than for shoots and lower for residues of perennial grasses than for barley. Cumulative respiration was positively correlated with water-soluble C in the residues but not with residue C/N. In the mixtures, the measured cumulative respiration was higher than the expected value in five of the nine mixes usually where the differences in cumulative respiration between the individual residues were relatively small. Lower than expected cumulative respiration were found in two of the mixtures in which barley shoots (high cumulative respiration) were mixed with residues with low cumulative respiration. There was a negative correlation between the change in microbial biomass C concentration from day 0 to day 18 and cumulative respiration on day 18. In the amended soils, the available N concentration decreased from day 0 to day 18. It is concluded that the low decomposition rate of perennial grasses residues should favour C sequestration, but that mixing residues of similar decomposition rate may accelerate their decomposition.Andong Shi, Chris Penfold, Petra Marschne