Oxidative Stress in Caenorhabditis elegans: Protective Effects of Spartin

Troyer syndrome is caused by a mutation in the SPG20 gene, which results in complete loss of expression of the protein spartin. We generated a genetic model of Troyer syndrome in worms to explore the locomotor consequences of a null mutation of the Caenorhabditis elegans SPG20 orthologue, F57B10.9, also known as spg-20. Spg-20 mutants showed decreased length, crawling speed, and thrashing frequency, and had a shorter lifespan than wild-type animals. These results suggest an age-dependent decline in motor function in mutant animals. The drug paraquat was used to induce oxidative stress for 4 days in the animals. We measured survival rate and examined locomotion by measuring crawling speed and thrashing frequency. After 4 days of paraquat exposure, 77% of wild-type animals survived, but only 38% of spg-20 mutant animals survived. Conversely, animals overexpressing spg-20 had a survival rate of 95%. We also tested lifespan after a 1 hour exposure to sodium azide. After a 24 hour recovery period, 87% of wild type animals survived, 57% of spg-20 mutant animals survived, and 82% of animals overexpressing spg-20 survived. In the behavioral assays, spg-20 mutant animals showed a significant decrease in both crawling speed and thrashing frequency compared with wild-type animals. Importantly, the locomotor phenotype for both crawling and thrashing was rescued in animals overexpressing spg-20. The animals overexpressing spg-20 had crawling speeds and thrashing frequencies similar to those of wild-type animals. These data suggest that the protein F57B10.9/SPG-20 might have a protective role against oxidative stress.     

A simple procedure for the isolation of streptomycin resistant plants in Solanaceae

A system has been developed for rapid selection of streptomycin resistant mutants, as adventitious shoots arising from explants of several Solanaceous species. Efficient mutagenesis was achieved by incubating shoot culture-derived leaf strips with 1 or 5 mM nitroso-methylurea, for 90 or 120 min. In Nicotiana tabacum and Lycopersicon peruvianum these treatments resulted in white or variegated adventitious shoots from up to 3.5% of explants placed on medium promoting shoot regeneration. Chlorophyll deficiencies were only observed very rarely in Solanum nigrum. Streptomycin resistant shoots were obtained from leaf explants placed on medium containing 500 mg l^-1 streptomycin sulphate, under which conditions explants are bleached and adventitious shoot development suppressed. Green adventitious s shoots appeared at a frequency dependent both on the mutagenic treatment and on the species. The best response was with S. nigrum where >70% of the explants produced streptomycin resistant shoots, most of which retained their resistance on subsequent testing. Maternal inheritance of streptomycin resistance has been confirmed for several N. tabacum and S. nigrum mutants, and there is also evidence for paternal transmission in the latter species. The procedure has been successfully extended to other species, including N. sylvestris and N. plumbaginifolia, and also to obtain spectinomycin resistant mutants.Communicated by R. Hagemann     

Microsatellite loci from the house wren (Troglodytes aedon)

We cloned seven microsatellite loci from house wrens (Troglodytes aedon) using a biotin enrichment protocol. Starting with fragments generated using DOP–PCR, fragments containing microsatellite motifs AC and AAC were captured using biotinylated probes and streptavidin coated magnetic particles. Captured fragments were cloned into plasmids; prior to sequencing, the plasmids were screened for microsatellites using a simple PCR approach. Five of the loci showed variation in a sample of nine individuals.     

Kurtzmanomyces tardus sp. nov., a new anamorphic yeast species of basidiomycetous affinity

A new yeast species of basidiomycetous affinity Kurtzmanomyces tardus was isolated from contaminated demineralized water. It differs from K. nectairii, the only other Kurtzmanomyces species so far described, in its carbon assimilation pattern and low DNA-DNA homology (2.3%±2.1).     

Maternal protein restriction induce skeletal muscle changes without altering the MRFs MyoD and myogenin expression in offspring

Stimuli during pregnancy, such as protein restriction, can affect morphophysiological parameters in the offspring with consequences in adulthood. The phenomenon known as fetal programming can cause short- and long-term changes in the skeletal muscle phenotype. We investigated the morphology and the myogenic regulatory factors (MRFs) MyoD and myogenin expression in soleus, SOL; oxidative and slow twitching and in extensor digitorum longus, EDL; glycolytic and fast twitching muscles in the offspring of dams subjected to protein restriction during pregnancy. Four groups of male Wistar offspring rats were studied. Offspring from dams fed a low-protein diet (6?% protein, LP) and normal protein diet (17?% protein, NP) were euthanized at 30 and 112?days old, and their muscles were removed and kept at ?80?°C. Muscles histological sections (8?μm) were submitted to a myofibrillar adenosine triphosphatase histochemistry reaction for morphometric analysis. Gene and protein expression levels of MyoD and myogenin were determined by RT-qPCR and western blotting. The major findings observed were distinct patterns of morphological changes in SOL and EDL muscles in LP offspring at 30 and 112?days old without changes in MRFs MyoD and myogenin expression. Our results indicate that maternal protein restriction followed by normal diet after birth induced morphological changes in muscles with distinct morphofunctional characteristics over the long term, but did not alter the MRFs MyoD and myogenin expression. Further studies are necessary to better understand the mechanisms underlying the maternal protein restriction response on skeletal muscle.     

Phytoplankton spring bloom in the Tagus coastal waters: hydrological and chemical conditions

Spatial variability of phytoplankton as well as hydrological and chemical conditions in the Tagus coastal waters were studied during the spring of 1994. The highly patchy distribution of phytoplankton and the community structure were related to the specific abiotic conditions prevailing in the area. Two main water masses were distinguished: a tidally mixed one alongshore Lisbon-cape Espichel and a stratified region strongly related to the Tagus plume river. The latter seems to control, to some extent, the spatial phytoplankton variability and the timing and size of the spring bloom. The highest surficial levels of nutrients, specifically PO₄ ^3–=1.00 M, NO₃ ^–=16.0 M and Si(OH)₄=14.6 M, were clearly associated with Tagus river discharges, while the lowest levels were attained offshore in the zone of maximum chlorophyll a concentrations (30 mg m^–3) reflecting the occurrence of high nutrient consumption. Furthermore, the near depletion of PO₄ ^3– indicates P limitation for phytoplankton growth. The diatom Detonula pumila (0.1×10⁶ cells l^–1) was the dominant species of phytoplankton communities in the mixed zone, alongshore Lisbon-cape Espichel, whereas, the small diatom Thalassiosira sp. (3×10⁶ cells l^–1) was blooming offshore, associated with the stratified region.     

Primary production and pigments in three low alkalinity connected reservoirs receiving mine wastes

A comparative study of primary production and pigments, conducted from April 1981 to April 1982, in a chain of three low alkalinity reservoirs situated in north of Portugal revealed significant lower values (159 mg C m⁻² d⁻¹ and 19.8 mg m⁻² as chlorophyll a, growing season means) in the reservoir receiving wastes rich in copper from a tungsten copper pyrite mine. This contrasts specially with the situation observed in the upstream reservoir which presents the highest values (409 mg C m⁻² d⁻¹ and 55.2 mg m⁻²) recorded in the system. Eventual effects of the mine discharge on phytoplankton biomass and photosynthesis such as high turbidity and copper toxicity are discussed. Results point out to be the presence of copper responsible for the alterations in the phytoplankton community. The system exhibits similar photosynthetic capacity which indicates an adaption to the relatively high copper concentrations in water and sediments. Experiments carried out in the largest unpolluted reservoir show that the loss of carbon through respiration is of major importance and indicate the phosphorus to be one of the factors limiting phytoplankton productivity.     

Temperature functions of thermal death in yeasts and their relation to the maximum temperature for growth

Summary Parameters of thermal death were determined in 10 strains of yeast species whose maximum temperatures for growth (T _max) ranged from 22 to 49°C. Arrhenius plots of the specific thermal death rates (k _d) formed a positional sequence at the level of the experimental points that corrresponded in all but one case to the sequence of the respective T _max values. Extrapolated k _d values at higher or lower temperatures no longer formed this sequence.The correlation of the temperature functions with T _max could be characterized in terms of a new activation parameter, for which the name thermal death activation constant is introduced. It has the following form: T.D.A. – S where H and S are respectively the apparent heat and entropy of activation of thermal death and n is the number of degrees above T _max (expressed in °K) at which the T.D.A. constant exists.Seven mesophilic yeasts had a T.D.A. constant between 72 and 79 calxmol^-1 degree^-1 at n values between 1 and 4°. This suggested that the destructive process that limits k _d in these strains is of the same species as one that contributes to the establishment of T _max. Two psychrophilic yeasts apparently had a similar T.D.A. constant but at a high n value (about 12.5°C) which suggested that in these strains T _max is governed by a destructive process unrelated to the one that underlies thermal death. The strain of the nearly thermophilic Hansenula angusta (T _max 49°C) did not fit in either group.The significance of the T.D.A. constant is discussed and expressions for H and S in terms of bond activation parameters are proposed.     

Multiple paternity in a salamander with socially monogamous behaviour

In the majority of birds and mammals, social monogamy is not congruent with genetic monogamy. No research to date has compared social and genetic monogamy in amphibians. We analysed paternity in clutches of red-backed salamanders (Plethodon cinereus), a species in which social monogamy has been demonstrated in the laboratory, and 28% of individuals in the forest are found in male-female pairs in the noncourtship season. We collected 16 clutches of eggs of P. cinereus in the southern Appalachian Mountains of Virginia and collected tail clippings from attending mothers. We genotyped embryos and adults at five microsatellite loci in order to analyse paternity of clutches. Most clutches (84.6%) had multiple sires, with two to three sires per clutch. In this study, 25% of clutches had males in addition to females attending eggs. None of the mothers of these clutches were genetically monogamous. All attending males sired some of the offspring in the clutch that they attended (between 9% and 50%) but never sired a majority in that clutch. We conclude that, at least in this population, social monogamy in P. cinereus is not concomitant with genetic monogamy.     

Interpreting population differentiation in terms of drift and selection

Summary Many empirical studies demonstrate some degree of genetic differentiation among populations of the same species. Understanding the relative importance of the processes causing this genetic differentiation has proven to be a difficult task. In particular, population differentiation can be influenced primarily by selection, genetic drift, and migration. We review the effect of drift and migration on patterns of genetic variation, with special reference to the conditions necessary for population differentiation. Conceptually, selection may be implicated in cases of population differentiation if the effect of drift and migration can be shown to be insufficient to cause the observed patterns. We examine some of the pitfalls of this approach when used with allozyme data, and revise a previous conclusion concerning the relative importance of selection in poulations of scale insects.