Canyon Wall and Floor Debris Deposits in Aeolis Mons, Mars.

Aeolis Mons (informally, Mount Sharp) exhibits a number of canyons, including Gediz and Sakarya Valles. Poorly sorted debris deposits are evident on both canyon floors and connect with debris extending down the walls for canyon segments that cut through sulphate-bearing strata. On the floor of Gediz Vallis, debris overfills a central channel and merges with a massive debris ridge located at the canyon terminus. One wall-based debris ridge is evident. In comparison, the floor of Sakarya Vallis exhibits a complex array of debris deposits. Debris deposits on wall segments within Sakarya Vallis are mainly contained within chutes that extend downhill from scarps. Lateral debris ridges are also evident on chute margins. We interpret the debris deposits in the two canyons to be a consequence of one or more late-stage hydrogeomorphic events that increased the probability of landslides, assembled and channelized debris on the canyon floors, and moved materials down-canyon. The highly soluble nature of the sulphate-bearing rocks likely contributed to enhanced debris generation by concurrent aqueous weathering to produce blocky regolith for transport downslope by fluvial activity and landslides, including some landslides that became debris flows. Subsequent wind erosion in Gediz Vallis removed most of the debris deposits within that canyon and partially eroded the deposits within Sakarya Vallis. The enhanced wind erosion within Gediz Vallis was a consequence of the canyon's alignment with prevailing slope winds.

Relationships between nitric oxide, nitroxyl ion, nitrosonium cation and peroxynitrite.

This review is concerned mainly with the three redox-related, but chemically distinct, species NO-, NO. and NO+, with greatest emphasis being placed on the chemistry and biology of the nitroxyl ion. Biochemical routes for the formation of nitroxyl ion and methods for showing the intermediacy of this species are discussed, together with chemical methods for generating nitroxyl ion in solution. Reactions of nitroxyl ion with NO., thiols, iron centres in haem and with dioxygen are reviewed The significance of the reaction between NO- and dioxygen as a source of peroxynitrite is assessed, and attention drawn to the possible significance of the spin state of the nitroxyl ion in this context. The biological significance of nitrosation and the importance of S-nitrosothiols and certain metal nitrosyl complexes as carriers of NO+ at physiological pH is stressed. Some features in the chemistry of peroxynitrite are noted.

Kinetics of the reactions of trioxodinitrate and nitrite ions with cytochrome d in Escherichia coli.

The rate of reaction of trioxodinitrate with reduced cytochrome oxidase d in membrane particles from Escherichia coli at pH 7 and 25 degrees C depends linearly upon [HN2O3-] over the concentration range studied (up to 0.05 mM) and is also first-order in cytochrome d. The known rate of decomposition of trioxodinitrate to give NO- and NO2- is about 4.5-times faster than the rate of reaction of reduced cytochrome d with trioxodinitrate, implying that cytochrome d reacts directly with NO-, with a trapping ratio of between 0.20 and 0.25, rather than with trioxodinitrate. The implications of the facile formation of the NO(-)-nitrosyl complex of cytochrome d for the mechanism of denitrification are discussed with particular reference to the mechanism of N-N bond formation. The reaction of reduced cytochrome d with nitrite (a decomposition product of trioxodinitrate) under these conditions is much slower than that with trioxodinitrate. The kinetics show a biphasic dependence of initial rate upon nitrite concentration. The rate data at low [NO2-] are consistent with saturation of a high affinity site for nitrite, having Vmax = 4.29.10(-9) M s-1 and Km = 0.034 mM. The existence of two binding sites for nitrite is consistent with the suggestion that the cytochrome bd complex contains two cytochrome d haems.

Nitrite and nitrosyl compounds in food preservation.

Nitrite is consumed in the diet, through vegetables and drinking water. It is also added to meat products as a preservative. The potential risks of this practice are balanced against the unique protective effect against toxin-forming bacteria such as Clostridium botulinum. The chemistry of nitrite, and compounds derived from it, in food systems and bacterial cells are complex. It is known that the bactericidal species is not nitrite itself, but a compound or compounds derived from it during food preparation. Of a range of nitrosyl compounds tested, the anion of Roussin's black salt [Fe4S3(NO)7]- was the most inhibitory to C. sporogenes. This compound is active against both anaerobic and aerobic food-spoilage bacteria, while some other compounds are selective, indicating multiple sites of action. There are numerous possible targets for inhibition in the bacterial cells, including respiratory chains, iron-sulfur proteins and other metalloproteins, membranes and the genetic apparatus.

Nitrate, but not silver, ions induce spectral changes in Escherichia coli cytochrome d.

The absorbance maximum (630 nm) of reduced cytochrome d in Escherichia coli membrane particles was diminished by 160 microM AgNO3 or NaNO3 and accompanied by the formation of a species with an absorption maximum at 640-645 nm. Nitrite, trioxodinitrate and nitric oxide elicited qualitatively similar, but faster, changes in the spectrum of cytochrome d, suggesting that formation of a nitrosyl complex may be involved in all cases. In direct contrast to an earlier report, silver ions (160 microM) were without effect on the alpha-bands of reduced cytochromes d, b or a 1.

Anoxic function for the Escherichia coli flavohaemoglobin (Hmp): reversible binding of nitric oxide and reduction to nitrous oxide.

The flavohaemoglobin Hmp of Escherichia coli is inducible by nitric oxide (NO) and provides protection both aerobically and anaerobically from inhibition of growth by NO and agents that cause nitrosative stress. Here we report rapid kinetic studies of NO binding to Fe(III) Hmp with a second order rate constant of 7.5 x 10(5) M(-1) s(-1) to generate a nitrosyl adduct that was stable anoxically but decayed in the presence of air to reform the Fe(III) protein. NO displaced CO bound to dithionite-reduced Hmp but, remarkably, CO recombined after only 2 s at room temperature indicative of NO reduction and dissociation from the haem. Addition of NO to anoxic NADH-reduced Hmp also generated a nitrosyl species which persisted while NADH was oxidised. These results are consistent with direct demonstration by membrane-inlet mass spectrometry of NO consumption and nitrous oxide production during anoxic incubation of NADH-reduced Hmp. The results demonstrate a new mechanism by which Hmp may eliminate NO under anoxic growth conditions.

Pituitary adenomas: long-term results for radiotherapy alone and post-operative radiotherapy.

PURPOSE: Analysis of prognostic factors and long-term results of treatment of pituitary adenomas. METHODS AND MATERIALS: The study involved a retrospective review of outcome in a series of 268 patients with pituitary adenomas, treated at the Queensland Radium Institute from January 1962 to December 1986. The study population included 108 patients treated with radiotherapy alone and 160 patients treated by surgery and post-operative radiotherapy. In each group, univariate and multivariate analyses were conducted of possible prognostic factors including age, sex, performance status, tumor type, tumor extent, visual disturbance, radiotherapy dose, and field size. RESULTS: For radiotherapy alone, the 10-year progression-free survival rate was 60%, and overall tumor control was obtained in 77%. Univariate analysis suggested that tumor type and radiotherapy field size were of prognostic significance. Multivariate analysis confirmed that Prolactinoma subtype and increasing radiotherapy field size were independently predictive of reduced progression-free survival. Long-term visual complications occurred in 1% of patients treated by radiotherapy alone. In patients treated by surgery and post-operative radiotherapy, the 10-year progression-free survival rate was 77%, and overall tumor control was achieved in 83%. Univariate analysis suggested that tumor type, completeness of surgical excision, and radiotherapy dose were predictive of outcome. However, on multivariate analysis, only the extent of surgical excision predicted prognosis independently. Long-term visual sequelae were noted in 3% of patients treated by surgery and post-operative radiotherapy. CONCLUSION: Both radiotherapy alone and post-operative radiotherapy are effective in long-term control of pituitary adenomas, and produce acceptably low complication rates.

Carbon monoxide-binding properties of the cytochrome bo quinol oxidase complex in Escherichia coli are changed by copper deficiency in continuous culture.

A strain of Escherichia coli having elevated levels of cytochrome bo and lacking the cytochrome bd quinol oxidase was grown in chemostat culture at low copper levels. Such cells had lowered levels of copper and of total cytochrome b. Cytochrome o concentration was unchanged when assayed by conventional CO difference spectroscopy, but apparently diminished by 80% in copper-deficient cells as determined by photodissociation of bound CO at 193 K. This is attributed to depletion of copper in the oxidase of copper-deficient cells, causing rapid recombination of photodissociated CO to haem O. CO recombination was also more sensitive to low intensities of actinic light in copper-depleted oxidase. The results illustrate a further similarity between the active sites of o- and aa3-type terminal oxidases.

Inhibition of the cytochrome bd-terminated NADH oxidase system in Escherichia coli K-12 by divalent metal cations.

Co(II), Zn(II) and Cd(II) ions inhibited NADH oxidase activity in membranes prepared from two cytochrome bo'-deficient mutants of Escherichia coli K-12 with the following order of potency: Zn(II) > Cd(II) >> Co(II). The degree of inhibition exhibited by these metal ions was not diminished in membranes which contained elevated levels of the cytochrome bd complex, suggesting that the most sensitive site precedes this complex in the aerobic respiratory chain. For each of the metal ions studied, inhibition was determined to be of the non-competitive type. Based upon the efficacy with which EDTA alleviated inhibition, Co(II), Zn(II) and Cd(II) ions are proposed to inhibit NADH oxidase activity by binding to at least two sites in the respiratory chain with significantly different affinities.

The bacteriocidal effects of transition metal complexes containing the NO+ group on the food-spoilage bacterium Clostridium sporogenes.

The chemical and molecular mechanism of toxicity of nitrite towards food-spoilage bacteria such as Clostridium botulinum or Clostridium sporogenes is not well understood. In order to discover the active species and explore its chemistry, a number of compounds related to nitrite were synthesized. Their bacteriocidal effects on C. sporogenes were investigated in Oxoid nutrient broth No. 2 growth medium at pH 7.0. Inhibition of cell growth, expressed as the concentration which causes 50% cell inhibition, was observed with nitrite at 10 mM, whereas [Fe4S3(NO)7]-(the anion of Roussin's black salt) and (Fe2(SCH2CH2OH)2(NO)4] (a water-soluble Roussin's red salt ester) were found to be effective at 0.001 mM and 0.005 mM, respectively, confirming previous reports that iron-sulphur-nitrosyl complexes are much more toxic to these organisms than nitrite itself. The nitroprusside anion, [Fe(CN)5NO]2- was found to be toxic at 0.030 mM and the corresponding chromium species, [Cr(CN)5NO]3-, at 0.1 mM. Therefore, on the basis of the number of NO groups present, the nitrosylcyano complexes are comparable in activity with the iron-sulphur-nitrosyl compounds. These results show that neither iron nor sulphur are essential for the bacteriostatic effect of the Roussin's type compounds. The property that all these compounds have in common is that they contain NO+. It is proposed that this is the active species responsible for the preservative effect of nitrite, and that a relationship may exist between the N-O stretching frequency, a measure of the NO+ character, and the toxicity of these NO(+)-containing complexes.

Metal ion-catalysed hydrolysis of ampicillin in microbiological growth media.

Anodic stripping voltammetry of bacterial growth medium containing copper(II) and ampicillin shows that Cu(II) is complexed by the antibiotic and that this complex decomposes to give Cu(II) complexes with ligands derived from ampicillin. At pH 7, substantial decomposition of ampicillin occurs over a few minutes, and even the very low levels of Cu(II) in Chelex-extracted medium are able effectively to catalyse the decomposition. The significance of this observation was shown during the screening of an Escherichia coli cosmid library for clones exhibiting increased resistance to Zn(II), Co(II) or Cd(II); the unexpected growth of the ampicillin-sensitive host E. coli strain on Luria-Bertani plates containing ampicillin and any of these metals was attributed to metal ion-catalysed decomposition of ampicillin. The instability of ampicillin (and other beta-lactam antibiotics) to metal ion-catalysed hydrolysis means that great care must be taken to ensure that such reactions do not occur in growth media. Furthermore, it is clear that double selection for resistance to ampicillin and metals such as Cu(II), Zn(II), Co(II) and Cd(II) is impossible.

Evidence for the transport of zinc(II) ions via the pit inorganic phosphate transport system in Escherichia coli.

A locus involved in zinc(II) uptake in Escherichia coli K-12 was identified through the generation of a zinc(II)-resistant mutant by transposon (Tn10dCam) mutagenesis. The mutation was located within the pitA gene, which encodes the low-affinity inorganic phosphate transport system (Pit). The pitA mutant accumulated reduced amounts of zinc(II) when exposed to 0.5-2.0 mM ZnSO(4) during growth in Luria-Bertani medium.

The toxicity of thallium(I) to cardiac and skeletal muscle.

The effects of (a) partial or complete replacement of K+ by Tl+ in saline perfusing isolated rat heart and diaphragm preparations and (b) pulse injections of high concentrations of Tl+ or K+, have been studied. The immediate effect of Tl+ resembles that of higher concentrations of K+ and may reflect its more rapid penetration into the tissue. Tl+ appears to replace K+ on a 1:1 basis to an extent dependent upon the relative abundance of the two cations in the perfusion solution. However, analysis of diaphragm preparations after perfusion with salines containing increasing Tl+ but constant [K+ + Tl+] showed a related and progressive increase in total cation content. This effect, which was not seen in the presence of constant high (normal) K+ concentrations, may reflect an increase of the intracellular space brought about by the thallium. Functional effects of Tl+ were (a) preferential block of the phrenic nerve or neuromuscular junction over the muscle fibre and (b) transient but marked acceleration of cardiac frequency following pulse injections, which may be of value in analysing the pacemaker mechanism of the heart. In both tissues Tl+ is eventually toxic and probably irreversibly so.

Filamentation of Escherichia coli K12 elicited by some monomeric, dimeric and trimeric complexes of ruthenium in various oxidation states.

A number of ruthenium complexes were tested for their ability to induce filamentation in Escherichia coli. These included monomeric and dimeric complexes with ruthenium in the II or III oxidation states, as well as mixed-valence complexes with ruthenium in the (II,III) oxidation states. In general, dimeric mixed-valence Ru(II,III) complexes were the most active class of compound, although some complexes of this type were relatively inactive. These were pyrazine- or bipyridyl-bridged complexes which are known to involve strong metal-ligand interaction, which stabilizes the Ru(II) oxidation state. Some Ru(III) complexes were also significantly active in induction of filamentous growth in E. coli. One of these was [Ru(NH3)5Cl]Cl2, which did not inhibit electron transport, Mg2+-ATPase activity or DNA synthesis in E. coli, but like [Ru2(NH3)6Br3]Br2 X H2O was a potent inhibitor of respiration-driven calcium transport in the organism. Filament-inducing activity of the complex was reduced in the presence of NaCl, but not in the presence of added Ca2+, ethanol, calcium pantothenate, or E. coli 'division promoting extract'. This behaviour is also similar to that of [Ru2(NH3)6Br3]Br2 X H2O. It is suggested that both complexes may induce filamentation in E. coli by a common mechanism, which may involve interference with calcium metabolism, or a wall or membrane target, rather than interaction with DNA.

Applications of electron paramagnetic resonance spectroscopy to study interactions of iron proteins in cells with nitric oxide.

Nitric oxide and species derived from it have a wide range of biological functions. Some applications of electron paramagnetic resonance (EPR) spectroscopy are reviewed, for observing nitrosyl species in biological systems. Nitrite has long been used as a food preservative owing to its bacteriostatic effect on spoilage bacteria. Nitrosyl complexes such as sodium nitroprusside, which are added experimentally as NO-generators, themselves produce paramagnetic nitrosyl species, which may be seen by EPR. We have used this to observe the effects of nitroprusside on clostridial cells. After growth in the presence of sublethal concentrations of nitroprusside, the cells show they have been converted into other, presumably less toxic, nitrosyl complexes such as (RS)2Fe(NO)2. Nitric oxide is cytotoxic, partly due to its effects on mitochondria. This is exploited in the destruction of cancer cells by the immune system. The targets include iron-sulfur proteins. It appears that species derived from nitric oxide such as peroxynitrite may be responsible. Addition of peroxynitrite to mitochondria led to depletion of the EPR-detectable iron-sulfur clusters. Paramagnetic complexes are formed in vivo from hemoglobin, in conditions such as experimental endotoxic shock. This has been used to follow the course of production of NO by macrophages. We have examined the effects of suppression of NO synthase using biopterin antagonists. Another method is to use an injected NO-trapping agent, Fe-diethyldithiocarbamate (Fe-DETC) to detect accumulated NO by EPR. In this way we have observed the effects of depletion of serum arginine by arginase. In brains from victims of Parkinson's disease, a nitrosyl species, identified as nitrosyl hemoglobin, has been observed in substantia nigra. This is an indication for the involvement of nitric oxide or a derived species in the damage to this organ.