Supported liquid membrane transport studies on Am(III), Pu(IV), U(VI) and Sr(II) using irradiated TODGA.

Transport behaviour of actinides viz. Am(3+), Pu(4+) and UO(2)(2+) was investigated from nitric acid feed conditions using PTFE (polytetrafluoroethylene) flat sheet supported liquid membranes (SLM) containing an irradiated solvent system comprising of N,N,N',N'-tetra-n-octyldiglycolamide (TODGA) as the carrier extractant and N,N-di-n-hexyloctanamide (DHOA) as the phase modifier. The present studies were carried out in order to understand the effect of irradiation on the long term reusability of the SLM and the decontamination behaviour in the absorbed dose range of 0-100 MRad. The studies using irradiated carrier included those with irradiated TODGA without any phase modifier and with 0.1M as well as 0.5M DHOA as the phase modifier. Transport behaviour of all the metal ions were found to be seriously affected with increasing radiation dose which was reflected in the decreasing percentage transport (%T) as well as permeability co-efficient (P) values. Though Sr(II) transport was quite significant with all the three unirradiated solvent systems, it was surprisingly low (<5%) when solvents exposed to 100 MRad dose were used in the SLM. Separation factors (S.F.) of the actinides over Sr(II) were calculated and were found to increase at higher radiation doses suggesting possibility of getting better decontamination on prolonged use of the supported liquid membrane system.

Processing of 3'-phosphoglycolate-terminated DNA double strand breaks by Artemis nuclease.

The Artemis nuclease is required for V(D)J recombination and for repair of an as yet undefined subset of radiation-induced DNA double strand breaks. To assess the possibility that Artemis acts on oxidatively modified double strand break termini, its activity toward model DNA substrates, bearing either 3'-hydroxyl or 3'-phosphoglycolate moieties, was examined. A 3'-phosphoglycolate had little effect on Artemis-mediated trimming of long 3' overhangs (> or =9 nucleotides), which were efficiently trimmed to 4-5 nucleotides. However, 3'-phosphoglycolates on overhangs of 4-5 bases promoted Artemis-mediated removal of a single 3'-terminal nucleotide, while at least 2 nucleotides were trimmed from identical hydroxyl-terminated substrates. Artemis also efficiently removed a single nucleotide from a phosphoglycolate-terminated 3-base 3' overhang, while leaving an analogous hydroxyl-terminated overhang largely intact. Such removal was completely dependent on DNA-dependent protein kinase and ATP and was largely dependent on Ku, which markedly stimulated Artemis activity toward all 3' overhangs. Together, these data suggest that efficient Artemis-mediated cleavage of 3' overhangs requires a minimum of 2 nucleotides, or a nucleotide plus a phosphoglycolate, 3' to the cleavage site, as well as 2 unpaired nucleotides 5' to the cleavage site. Shorter 3'-phosphoglycolate-terminated overhangs and blunt ends were also processed by Artemis but much more slowly. Consistent with a role for Artemis in repair of terminally blocked double strand breaks in vivo, human cells lacking Artemis exhibited hypersensitivity to x-rays, bleomycin, and neocarzinostatin, which all induce 3'-phosphoglycolate-terminated double strand breaks.

Electron magnetic resonance study of gamma-irradiated poly(lactide-co-glycolide) microspheres.

A series of poly(lactide-co-glycolide) samples of compositions ranging from (75:25) to (65:35) to (50% (65:35):50% (50:50)) were gamma-irradiated under ambient conditions in air. The irradiation doses used were 15, 20, 25, and 30 kGy. The generation of radicals resulting from the gamma-irradiation was confirmed using EPR. Two major radical species were observed and identified as centered at alkyl and alkyl peroxy groups. The indication from the X-band (9 GHz) frequencies showed that alkyl radicals gave rise to a quartet hyperfine pattern. However, measurements performed at higher W-band frequencies (90 GHz) showed that the X-band spectra are actually a composite profile arising from a series of overlapping individual resonances. Using combined EPR and ENDOR (Electron-Nuclear DOuble Resonance) measurements, an alkyl peroxy radical was identified. For increasing glycolide concentration from 75:25 to the 50:50 blend, there was a factor of 7 increase in the concentration of radicals A and B. Furthermore, both radical species were found to be stable for several weeks after storage at ambient temperature conditions. At elevated temperatures and humidities, radical stability decreased--the decay rate was estimated at approximately 3x10(-8) mol K(-1). The stability characteristics of the radicals under different conditions are attributed to changes in the morphology of the polymer.

Poly(lactide-co-glycolide) microspheres containing bupivacaine: comparison between gamma and beta irradiation effects.

The beta- and gamma-irradiation effects on stability of microspheres made of poly(lactide-co-glycolide) 50:50 copolymer (PLGA) containing bupivacaine (BU) were studied. Microspheres containing 10, 25, and 40% w/w, respectively, of BU were prepared by spray drying and irradiated in air with beta- and gamma-irradiation at a dose of 25 kGy. Morphology (atomic force microscopy, particle-size analysis), physico-chemical characteristics (DSC and FT-IR spectroscopy), drug content and in vitro dissolution profile of microspheres were all determined; the stability of irradiated microspheres was evaluated over a 9-month period. The decrease of BU content in gamma-irradiated microspheres was almost always constant independent of the amount of BU per sample, therefore it was in inverse proportion to drug loading (range between 5 and 15%). BU release rate increased immediately after irradiation and increased slightly until 90 days of storage. As far as beta-irradiated microspheres are concerned, BU content decreased in a significant way (approximately 3%) only in microspheres containing 10% w/w of BU. Immediately after irradiation, drug release rate in beta-irradiated microspheres increased less than in the corresponding gamma-irradiated microspheres, and it did not change further over the following storage period. BU-loaded microspheres have been shown to be more stable against beta- than gamma-irradiation. AFM revealed that the surface roughness of the irradiated microspheres increases depending on irradiation. As such, if a parameter is quantifiable, it is proposed as a marker of degradation due to ionizing radiation.

Proton concentration jumps and generation of transmembrane pH-gradients by photolysis of 4-formyl-6-methoxy-3-nitrophenoxyacetic acid.

Proton concentration gradients across membranes are important for many biological energy transducing processes. The kinetics of proton dependent processes can be studied by pH-jump methods in which protons are photochemically released. In the following we describe the synthesis and the properties of photolabile 4-formyl-6-methoxy-3-nitrophenoxyacetic acid, a 'caged proton'. The synthesis is based on vanillin, which is alkylated with chloroacetic acid to give a carboxylic acid (pK = 2.72). In a second step a nitro group ortho to the formyl group is introduced. Photochemical proton release occurs by a reaction mechanism analogous to the well known photochemical formation of 2-nitrosobenzoic acid from 2-nitrobenzaldehyde. The pK values of the photoproduct are 0.75 and 2.76, respectively, thus allowing the use of the compound in a wide pH-range. The quantum yield is 0.18, lower than in the case of the 2-nitrobenzaldehyde/2-nitrosobenzoic acid system (phi = 0.5). The release of the proton in a flash photolysis experiment occurs within less than 1 microseconds. The spectrum of photolabile compound has absorption maxima at 263 nm and 345 nm, respectively. Its permeability across a lipid bilayer membrane is very low (permeability coefficient Pd approximately equal to 10(-9) cm.s-1 at pH 8) so that transmembrane proton concentration gradients can be generated.