One-electron Oxidation of a Pyrenyl Photosensitizer Covalently Attached to DNA and Competition Between its Further Oxidation and DNA Hole Injection

The photosensitized hole injection and guanine base damage phenomena have been investigated in the DNA sequence, 5′-d(CATG[subscript 1] [superscript Py]CG[subscript 2]TCCTAC) with a site-specifically positioned pyrene-like (Py) benzo[a]pyrene 7,8-diol 9,10-epoxide-derived N[superscript 2]-guanine adduct (G[subscript 1] [superscript Py]). Generation of the Py radical cation and subsequent hole injection into the DNA strand by a 355 nm nanosecond laser pulses (∼4 mJ cm[superscript −2]) results in the transformation of G[subscript 1] [superscript Py] to the imidazolone derivative Iz[subscript 1] [superscript Py] and a novel G[subscript 1] [superscript Py*] photoproduct that has a mass larger by 16 Da (M+16) than the mass (M) of G[subscript 1] [superscript Py]. In addition, hole transfer and the irreversible oxidation of G[subscript 2], followed by the formation of Iz[subscript 2] was observed (Yun et al. [2007], J. Am. Chem. Soc., 129, 9321). Oxygen-18 and deuterium isotope labeling methods, in combination with an extensive analysis of the MS/MS fragmentation patterns of the individual dG[superscript Py*] nucleoside adduct and other data show that dG[superscript Py*] has an unusual structure with a ruptured cyclohexenyl ring with a carbonyl group at the rupture site and intact guanine and pyrenyl residues. The formation of this product competes with hole injection and thus diminishes the efficiency of oxidation of guanines within the oligonucleotide strand by at least 15% in comparison with that in the dG[superscript Py] nucleoside adduct.National Institutes of Health (U.S.) (Grant R01 CA110261

Combustion Joining of Regolith Tiles for In-Situ Fabrication of Launch/Landing Pads on the Moon and Mars

To mitigate dust problems during launch-landing operations in lunar and Mars missions, it is desired to build solid pads on the surface. Recently, strong tiles have been fabricated from lunar regolith simulants using high-temperature sintering. The present work investigates combustion joining of these tiles through the use of exothermic intermetallic reactions. Specifically, nickel aluminum (1:1 mole ratio) mixture was placed in a gap between the tiles sintered from JSC-1A lunar regolith simulant. Upon ignition by a laser, a self-sustained propagation of the combustion front over the mixture occurred. Joining was improved with increasing the tile thickness from 6.3 mm to 12.7 mm. The temperatures sufficient for melting the glass phase of JSC-1A were recorded for 12.7-mm tiles, which explains the observed better joining

The Mars Hopper: Development, Simulation and Experimental Validation of a Radioisotope Exploration Probe for the Martian Surface

An advanced exploration probe has been proposed by the Center for Space Nuclear Research (CSNR) to acquire detailed data from the Martian surface and subsurface, ‘hop’ large distances to multiple sites in short periods of time and perform this task repeatedly. Although several similar flying vehicles have been proposed utilizing various power sources and complex designs, e.g. solar-electric and chemical-based, the CSNR’s Mars Hopper is based on a radioisotope thermal rocket (RTR) concept. The Mars Hopper’s design relies on the high specific energies [J/kg] of radioisotopes and enhances their low specific power [W/kg] through the use of a thermal capacitance material to store thermal energy over time. During operation, the RTR transfers the stored thermal energy to a flowing gas, which is then expanded through a converging-diverging nozzle, producing thrust. Between flights, the platform will have ample time to perform in-depth science at each location while the propellant tanks and thermal capacitor recharge. Recharging the propellant tanks is accomplished by sublimation freezing of the ambient CO2 atmosphere with a cryocooler, followed by heating and pressurization to yield a liquid storage state. The proposed Mars Hopper will undergo a ballistic flight, consuming the propellant in both ascent and descent, and by using multiple hopper platforms, information can be gathered on a global scale, enabling better resource resolution and providing valuable information for a possible Mars sample-return mission. The CSNR, collaborating with the Idaho National Laboratory (INL) and three universities (University of Idaho, Utah State University and Oregon State University), has identified key components and sub-systems necessary for the proposed hopper. Current project activities include the development of a lab-scale prototypic Mars Hopper and test facility, along with computational fluid dynamics (CFD)/thermal-hydraulic models to yield a better understanding of the heat transfer process and complex nature of turbulent CO2 flow. Laboratory experimentation will aid design iterations and the development of both tethered and free-flying terrestrial hoppers that utilize an electrically heated core. The knowledge base acquired from these activities will refine the Mars Hopper’s future performance and optimize the RTR core components prior to constructing the final design

The effects of nitroxyl (HNO) on soluble guanylate cyclase activity: interactions at ferrous heme and cysteine thiols

It has been previously proposed that nitric oxide (NO) is the only biologically relevant nitrogen oxide capable of activating the enzyme soluble guanylate cyclase (sGC). However, recent reports implicate HNO as another possible activator of sGC. Herein, we examine the affect of HNO donors on the activity of purified bovine lung sGC and find that, indeed, HNO is capable of activating this enzyme. Like NO, HNO activation appears to occur via interaction with the regulatory ferrous heme on sGC. Somewhat unexpectedly, HNO does not activate the ferric form of the enzyme. Finally, HNO-mediated cysteine thiol modification appears to also affect enzyme activity leading to inhibition. Thus, sGC activity can be regulated by HNO via interactions at both the regulatory heme and cysteine thiols

Oxidation of single-stranded oligonucleotides by carbonate radical anions: generating intrastrand cross-links between guanine and thymine bases separated by cytosines

The carbonate radical anion is a biologically important one-electron oxidant that can directly abstract an electron from guanine, the most easily oxidizable DNA base. Oxidation of the 5′-d(CCTACGCTACC) sequence by photochemically generated CO3·− radicals in low steady-state concentrations relevant to biological processes results in the formation of spiroiminodihydantoin diastereomers and a previously unknown lesion. The latter was excised from the oxidized oligonucleotides by enzymatic digestion with nuclease P1 and alkaline phosphatase and identified by LC-MS/MS as an unusual intrastrand cross-link between guanine and thymine. In order to further characterize the structure of this lesion, 5′-d(GpCpT) was exposed to CO3·− radicals, and the cyclic nature of the 5′-d(G*pCpT*) cross-link in which the guanine C8-atom is bound to the thymine N3-atom was confirmed by LC-MS/MS, 1D and 2D NMR studies. The effect of bridging C bases on the cross-link formation was studied in the series of 5′-d(GpCnpT) and 5′-d(TpCnpG) sequences with n = 0, 1, 2 and 3. Formation of the G*-T* cross-links is most efficient in the case of 5′-d(GpCpT). Cross-link formation (n = 0) was also observed in double-stranded DNA molecules derived from the self-complementary 5′-d(TTACGTACGTAA) sequence following exposure to CO3·− radicals and enzymatic excision of the 5′-d(G*pT*) product

Purdue Hydrogen Technology Program

Presented at the 2006 DOE Hydrogen, Fuel Cells & Infrastructure Technologies Program Annual Merit Review in Washington, D.C., May 16-19, 2006