Damage induced by 1-30 eV electrons on thymine- and bromouracil-substituted oligonucleotides.

The impact of low-energy (1-30 eV) electrons on self-assembled monolayers of heterogeneous oligonucleotides chemisorbed on a gold surface has been investigated by mass spectrometry of desorbed neutral species in an attempt to understand the consequences of secondary electron damage in a short sequence of a DNA single strand. We demonstrate that the most intense observable neutral species (CN, OCN and/or H(2)NCN) desorbed from Cy(6)-Th(3) and Cy(6)-(BrdU)(3) oligos are related to primary fragmentation of the bases induced by electron impact. The dependence of the neutral species desorption on electron energy shows typical signatures of dissociative electron attachment initiated by the formation of shape- and core-excited resonances (i.e. single-electron and two-electron- one-hole transitory anions, respectively). Substitution of dTh by BrdU increases the production of neutral fragments by as much as a factor of about 3 for the entire electron energy range. When the distribution of secondary electrons along radiation tracks in H(2)O is taken into account, we show that the probability for electron damage to heterogeneous oligonucleotides is enhanced by a factor of 2.5-3 for electron energies below 20 eV for both sensitized and unsensitized strands.

Low-energy (5-25 eV) electron damage to homo-oligonucleotides.

Radiation-induced damage to homo-oligonucleotides is investigated by electron-stimulated desorption of neutral fragments from chemisorbed organic films. Six and 12 mers of cytidine phosphate (poly dCs) and thymidine phosphate (poly dTs) are chemisorbed from various solutions onto a crystalline gold substrate by a thiol modification at the 3' end and are irradiated under ultra-high vacuum conditions with 5-25 eV electrons. The mass selected neutral desorption yields consist mainly of fragments of the DNA bases, i.e. CN and OCN (and/or H2NCN for poly dCs) from both poly dCs and poly dTs, indicating that the electrons interact specifically via fragmentation of the aromatic ring of either of the bases. Other heavier fragments are also detected such as H3CC-CO from poly dTs. The yields generally possess a threshold near 5 eV and a broad maximum around 12-13 eV incident electron energy. Dissociative electron attachment as well as electronically excited neutral or cation states are believed to be responsible for the various desorption yields. The latter yields are consistently larger for oligos chemisorbed from water and acetone solutions, compared to methanol solution. The invariance of the fragment yield intensities with oligo length suggests that the molecules are likely to adsorb almost parallel to the surface.

Pentoxifylline suppression of TNF-alpha mediated axonal degeneration in the rabbit optic nerve.

In AIDS patients, axonal degeneration in the optic nerve occurs as a histopathological manifestation of the optic neuropathy. Direct infection of neurons by HIV is unlikely, and the axonal injury may be an indirect effect mediated by cytotoxic factors such as tumor necrosis factor-alpha (TNF-alpha) which we have previously demonstrated to cause axonal degeneration in the rabbit optic nerve. To test the suppressive effects of pentoxifylline in preventing TNF-alpha-mediated axonal degeneration, we applied pentoxifylline to an established rabbit model that demonstrates an AIDS-like optic neuropathy using intravitreal TNF-alpha injections. Degenerated axonal profiles were numerous in control rabbit optic nerve (mean 1879) and reduced in rabbits receiving the medium dose of pentoxifylline (300 mg PO BID, mean 439, p < 0.001) and the highest dose of pentoxifylline (600 mg PO BID, mean 120, p < 0.007). High dose pentoxifylline reduced TNF-alpha-induced axonal losses to less than 10% that seen without pentoxifylline pretreatment. Lower doses of pentoxifylline had a lesser but significant protective effect. Our results suggest that TNF-alpha-mediated axonal degeneration can be suppressed by high doses of pentoxifylline. Pentoxifylline may therefore be useful in AIDS patients demonstrating neurological or neuro-ophthalmological symptoms.

A test for patency of the cystic duct in acute cholecystitis.

A procedure was devised to quickly and reliably determine the patency of the cystic duct in patients suspected of having acute cholecystitis. First the gallbladder was stimulated to empty by a cholecystokinin injection. Thirty minutes later a radiolabeled biliary marker, either 150 muCi 131-I rose bengal or 2 mCi 99-mTc dihydrothioctic acid, was injected, and the accumulation of radioactivity in the liver and gallbladder regions was monitored by external gamma emission imaging and recording devices. The images of diagnostic importance were obtained between 60 and 90 minutes after injection of the tracer. Thirty-nine patients with acute abdominal pain were studied. Ten patients who had acute cholecystitis failed to show gallbladder accumulation of radioactivity, reflecting the cystic duct obstruction that initiates this disease. Twenty-nine patients having a variety of other diseases all showed gallbladder accumulation of activity, indicating in each patient that the cystic duct was patent. No significant adverse effects were noted. We conclude that the procedure is a useful adjunct to the clinical and roentgenographic evaluation of patients with acute abdominal pain.