Transport of amino acids and ammonium in mycelium of Agaricus bisporus.

Mycelium of Agaricus bisporus took up methylamine (MA), glutamate, glutamine and arginine by high-affinity transport systems following Michaelis-Menten kinetics. The activities of these systems were influenced by the nitrogen source used for mycelial growth. Moreover, MA, glutamate and glutamine uptakes were derepressed by nitrogen starvation, whereas arginine uptake was repressed. The two ammonium-specific transport systems with different affinities and capacities were inhibited by NH(+)(4), with a K(i) of 3.7 microM for the high-velocity system. The K(m) values for glutamate, glutamine and arginine transport were 124, 151 and 32 microM, respectively. Inhibition of arginine uptake by lysine and histidine showed that they are competitive inhibitors. MA, glutamate and glutamine uptake was inversely proportional to the intracellular NH(+)(4) concentration. Moreover, increase of the intracellular NH(+)(4) level caused by PPT (DL-phosphinotricin) resulted in an immediate cessation of MA, glutamine and glutamate uptake. It seems that the intracellular NH(+)(4) concentration regulates its own influx by feedback-inhibition of the uptake system and probably also its efflux which becomes apparent when mycelium is grown on protein. Addition of extracellular NH(+)(4) did not inhibit glutamine uptake, suggesting that NH(+)(4) and glutamine are equally preferred nitrogen sources. The physiological importance of these uptake systems for the utilization of nitrogen compounds by A. bisporus is discussed.     

Determination of deuterium-labeled tryptophan in proteins by means of high-performance liquid chromatography and thermospray mass spectrometry

In order to develop direct methods for determining the extent of metabolic incorporation of isotopically labeled amino acids into a protein, the determination of deuterated tryptophan in [2H5]tryptophan-bacteriorhodopsin was investigated. The isotopically modified protein was subjected to alkaline hydrolysis. After phenyl isothiocyanate derivatization of the hydrolysate, the mixture was separated by reversed-phase liquid chromatography. Field desorption mass spectrometry and thermospray mass spectrometry were investigated for their ability to determine the ratio between [2H5]tryptophan and total tryptophan in the collected fractions. In order to check the procedure a set of known tryptophan/[2H5]tryptophan mixtures were passed through the same derivatization, HPLC separation, and lyophilization procedure as used for the biological samples.     

Evolution of sprouting versus seeding inAspalathus linearis

We have tested the hypothesis that reseeding is a plesiomorphic character state and that sprouting is a derived state inAspalathus linearis, and that the latter is an adaptation to ensure fire-survival in a fireprone environment. Samples of five seeder and four sprouter populations of A.linearis were examined by horizontal starch gel electrophoresis to assess the amount of genetic differentiation within and between sprouting and seeding populations, and to determine the extent of gene flow between the populations. Leaf extracts were surveyed for ten enzymes and gene products revealed genetic variation at 13 (76%) of 17 protein coding loci. Allele frequency differences were found between sprouting and seeding populations and genetic distance values show that the sprouters are grouped separate from the seeders, thus providing support for the morphological data on which the above mentioned hypothesis is based. It is evident that evolution operates at the population level inA. linearis.     

Pycnogenol® and vitamin E inhibit ethanol‐induced apoptosis in rat cerebellar granule cells

Pycnogenol® (PYC), a patented combination of bioflavonoids extracted from the bark of French maritime pine (Pinus maritima), scavenges free radicals and promotes cellular health. The protective capacity of PYC against ethanol toxicity of neurons has not previously been explored. The present study demonstrates that in postnatal day 9 (P9) rat cerebellar granule cells the antioxidants vitamin E (VE) and PYC (1) dose dependently block cell death following 400, 800, and 1600 mg/dL ethanol exposure (2) inhibit the ethanol‐induced activation of caspase‐3 in the same model system; and (3) reduce neuronal membrane disruption as assayed by phosphatidylserine translocation to the cell surface. These results suggest that both PYC and VE have the potential to act as therapeutic agents, antagonizing the induction of neuronal cell death by ethanol exposure. © 2004 Wiley Periodicals, Inc. J Neurobiol 59: 261–271, 2004     

Prey-capture techniques and prey preferences of Corythalia canosa and Pystira orbiculata, ant-eating jumping spiders (Araneae, Salticidae)

Corythalia canosa from Florida is an unusual salticid because it is known to eat ants. This species' specialized behaviour for catching ants is described in detail for the first time and compared to its behaviour for catching other insects. Pystira orbiculata from Queensland is shown to be another ant-eating salticid, although its behaviour for catching ants seems less specialized than that of C. canosa. Three different types of tests of prey preference were carried out. In each type of test C. canosa and P. orbiculata took ants in preference to other insects. Another species of salticid, Trite planiceps from New Zealand, failed to eat ants in these tests, although T. planiceps often attacked then released the ants. Corythalia canosa's and P. orbiculata's preference for ants, and their prey-specific predatory behaviour for catching ants, are shown not to depend on prior experience with ants.