Oxygen-dependent antagonism of lipid peroxidation

Measurements of the rates for formation of conjugated dienes, malonylaldehyde, and lipid hydroperoxides show that increasing the concentration of O2 from 0.11 mM to 0.35 mM or 0.69 mM can slow the rate of linoleic acid peroxidation in a xanthine oxidase/hypoxanthine system. This effect is seen at pH 7.0 but not 7.4 and depends on the presence of monounsaturated fatty acids (oleic, cis, or trans vaccenic acid). Oxygen antagonism of ascorbic acid-iron-EDTA mediated lipid peroxidation is similarly dependent on fatty acid mixtures and occurs at pH 5.0 and 6.0 but not 7.0. The efficiency of initiation of peroxidation in the xanthine oxidase system is unaffected by monounsaturated fatty acids and O2 concentration. Increasing the O2 concentration increases the rate of superoxide radical production, but there is no change in salicylate hydroxylation (e.g., OH. production) or ferrous ion concentration. Oxygen-mediated slower rates of lipid peroxidation are associated with either increased H2O2 production or, based on an indirect assay, singlet O2 production. Increased O2 concentrations increase the rate of azobisisobutyronitrile-initiated lipid peroxidation as expected but addition of exogenous superoxide radicals slows the rate. Under similar conditions superoxide reacts with fatty acids to produce singlet O2. Overall, the data suggest that O2-mediated antagonism occurs because of termination reactions between hydroperoxyl (HO2.) and organic radicals, and singlet O2 or H2O2 are products of these reactions.     

Floristic Development Patterns in a Restored Elk River Estuarine Marsh, Grays Harbor, Washington

We describe the changes in the floral assemblage in a salt marsh after reconnection to estuarine tidal inundation. The Elk River marsh in Grays Harbor, Washington was opened to tidal flushing in 1987 after being diked for approximately 70 years. The freshwater pasture assemblage dominated by Phalarais arundinacea (reed canary grass) converted to low salt marsh vegetation within 5 years, with the major flux in species occurring between years 1 and 4. The system continued to develop through the 11‐year post‐breach monitoring period, although change after year 6 was slower than in previous years. The assemblage resembles a low salt marsh community dominated by Distichlis spicata (salt grass) and Salicornia virginica (pickleweed). Because of subsidence of the system during the period of breaching, the restored system remains substantially different from the Deschamsia cespitosa (tufted hairgrass)‐dominated reference marsh. Use of a similarity index to compare between years and also between reference and restored marshes in the same year revealed that similarity in floral composition between year 0 and subsequent years decreased with time. However, there was a period of dramatic dissimilarity during years 1 to 3 when the system was rapidly changing from a freshwater to estuarine condition. Similarity values between the reference and restored system generally increased with time. Somewhat surprisingly the reference marsh showed considerable between‐year variation in similarity, which indicated substantial year‐to‐year variability in species composition. Based on accretion rate data from previous studies we predict that full recovery of the system would take between 75 and 150 years.     

Female American mink, Mustela vison, mate multiply in a free-choice environment

Like most solitary carnivores, the home ranges of male American mink overlap with those of several females, but each female typically shares its range with only a single male. Nevertheless, female mink produce multiply sired litters. Unusually among mammals, the physiological characteristics of female American mink make them highly suited to multiple paternity, suggesting that polyandry in the species may result from female behaviour rather than from coercion by males. In a free-choice experiment on captive-bred animals, all female mink mated multiply, with seven of the eight mating with all three available partners. Since females sometimes resisted copulations, multiple mating in this species may result from their behaviour rather than male coercion. Females visited males nonrandomly, but visiting patterns did not predict mating patterns, suggesting disparate social and mating preferences. There was no evidence for precopulatory female choice, since the allocation of copulations among available males did not deviate significantly from the expected distribution. Females mated multiply only around the predicted times of ovulation, contrary to the expectation that they should be more selective at times of peak fertility. Larger males copulated for longer in total: copulation duration is probably under male control, suggesting that larger males could influence mating duration, perhaps increasing their share of paternity by greater sperm transfer. Our observations suggest that in wild populations, mating patterns could be influenced by female preference for multiple partners.     

Fluorescence properties of reduced thionicotinamide--adenine dinucleotide and of its complex with octopine dehydrogenase.

Reduced 3-thionicotinamide--adenine dinucleotide (sNADH) is shown to be fluorescent, with an emission maximum at 510 nm when excited in the region of the absorption maximum (398 nm), and with a very low quantum yield, (3.4 +/- 0.5) x 10(-4). The interaction between sNADH and octopine dehydrogenase was investigated by ultraviolet-difference spectroscopy and fluorescence. Some surprising fluorescence features were found when sNADH was bound to the enzyme in the presence of D-octopine, as follows. (a) There is an unusually high enhancement of the dinucleotide fluorescence (by at least a factor of 100) attended by a 40-nm blue shift of the emission maximum. (b) The protein fluorescence is quenched almost completely. (c) The bound coenzyme analog undergoes a photoreaction, which proceeds differently from that occurring the free form. These features appear to be unique to the octopine.sNADH complex, as for example they are not present when sNADH is bound to horse liver alcohol dehydrogenase, or when NADH is bound to octopine dehydrogenase. The possible origin of these fluorescence features is discussed. Binding and kinetic studies were carried out with sNAD and sNADH. It was found that sNAD neither binds nor acts kinetically as a coenzyme. sNADH exhibits relatively good binding, with Km and Ki values close to those of the natural coenzyme, but the turnover number is 460 times smaller than that with NADH. The kinetic consequences of these findings are discussed. The sNADH dissociation constants were determined as a function of temperature, and appear to be practically temperature-independent in the range 10--40 degrees C. It seems thus, in agreement with previous studies, that the interaction between octopine dehydrogenase and coenzymes proceeds athermically, regardless of the structure, affinity, and chemical reactivity of the coenzyme. The possible biological and chemical meaning of this finding is discussed.     

Reactions of nucleic acid bases with alpha-acetylenic esters. Synthesis and enzymatic evaluation of chemically modified nucleotides

Analogues of adenine nucleotides, containing an additional chloromethyl-pyrimidone ring fused to the purine base, were obtained by treatment of AMP, ADP and ATP with an alpha-acetylenic ester, methyl 4-chlorobut-2-ynoate. These compounds were tested for their ability to substitute for the natural substrates or cofactors of several enzymes. With the ADP analogue, pyruvate kinase showed a significant increase of the Km value and a moderate decrease of V, while the reverse was observed when hexokinase was tested with modified ATP. Adenylate kinase was active with the ATP analogue but not with the AMP derivative. Myosin accepted the ATP analogue as a substrate, but was irreversibly modified. Among the dehydrogenases tested, only glucose-6-phosphate dehydrogenase was inhibited by the nucleotide analogue. The structure--activity relationship of these nucleotide derivatives, which represent the largest dimensional deviation known from natural nucleotides, is discussed in comparison with some earlier described dimensional probes of enzyme-nucleotide binding sites.     

Intra- and interspecific variation in the chitin content of some anostracans

Chitin was assayed in adults of nine anostracan species (Streptocephalus dichotomus, S. proboscideus, S. simplex, S. torvicornis, Thamnocephalus platyurus, Dendrocephalus brasiliensis, Chirocephalus diaphanus, Artemia salina and Branchipus schaefferi) using a highly specific enzymatic method. Cysts of A. salina, T. platyurus and larvae of A. salina and S. simplex were also analysed for their chitin content. The results demonstrated a weak interspecific variation in chitin content among the adults. On the whole, it ranged from 9 to 33 mg chitin (g dry mass)⁻¹. In most species, the chitin level was not significantly different between males and females. In contrast, chitin levels varied significantly between cysts, larvae and adults within species. Comparison of the chitin content of anostracans revealed a comparable or lower chitin level than other crustaceans. This revised version was published online in August 2006 with corrections to the Cover Date.