A developmentally regulated neuraminidase activity in Trypanosoma cruzi

The human pathogen Trypanosoma cruzi (Y strain) contains a neuraminidase activity that varies widely in the different developmental stages of the parasite. The specific neuraminidase activity of infective trypomastigotes obtained from tissue culture and from the bloodstream of infected mice is 7 to 15 times higher than that of the acellular culture forms. Amastigotes were devoid of enzyme activity. The enzyme has a pH optimum of 6.0 to 6.5. Live trypanosomes released sialic acid from human erythrocytes and plasma glycoproteins. Several sialyl compounds were hydrolyzed by the parasite, but the best substrate was the protein orosomucoid. Erythrocytes from infected mice with T. cruzi parasitemia were agglutinated by peanut lectin and the hemagglutination titer was correlated with the degree of parasitemia.     

Essential role of Fkbp6 in male fertility and homologous chromosome pairing in meiosis

Meiosis is a critical stage of gametogenesis in which alignment and synapsis of chromosomal pairs occur, allowing for the recombination of maternal and paternal genomes. Here we show that FK506 binding protein (Fkbp6) localizes to meiotic chromosome cores and regions of homologous chromosome synapsis. Targeted inactivation of Fkbp6 in mice results in aspermic males and the absence of normal pachytene spermatocytes. Moreover, we identified the deletion of Fkbp6 exon 8 as the causative mutation in spontaneously male sterile as/as mutant rats. Loss of Fkbp6 results in abnormal pairing and misalignments between homologous chromosomes, nonhomologous partner switches, and autosynapsis of X chromosome cores in meiotic spermatocytes. Fertility and meiosis are normal in Fkbp6 mutant females. Thus, Fkbp6 is a component of the synaptonemal complex essential for sex-specific fertility and for the fidelity of homologous chromosome pairing in meiosis.     

Transgenic RNAi reveals essential function for CTCF in H19 gene imprinting

The imprinted regulation of H19 and Insulin-like growth factor 2 expression involves binding of the vertebrate insulator protein, CCCTC binding factor (CTCF), to the maternally hypomethylated differentially methylated domain (DMD). How this hypomethylated state is maintained during oogenesis and the role of CTCF, if any, in this process are not understood. With the use of a transgenic RNA interference (RNAi)-based approach to generate oocytes with reduced amounts of CTCF protein, we found increased methylation of the H19 DMD and decreased developmental competence of CTCF-deficient oocytes. Our results suggest that CTCF protects the H19 DMD from de novo methylation during oocyte growth and is required for normal preimplantation development.     

Synthetic clonal reproduction through seeds

Cloning through seeds has potential revolutionary applications in agriculture, because it would allow vigorous hybrids to be propagated indefinitely. However, asexual seed formation or apomixis, avoiding meiosis and fertilization, is not found in the major food crops. To develop de novo synthesis of apomixis, we crossed Arabidopsis MiMe and dyad mutants that produce diploid clonal gametes to a strain whose chromosomes are engineered to be eliminated after fertilization. Up to 34% of the progeny were clones of their parent, demonstrating the conversion of clonal female or male gametes into seeds. We also show that first-generation cloned plants can be cloned again. Clonal reproduction through seeds can therefore be achieved in a sexual plant by manipulating two to four conserved genes.     

Pericarp total protein profiles as molecular markers of tomato fruit quality traits in two segregating populations

The tomato fruit quality results of biochemical and physiological changes that occur during the ripening process. Although, the pericarp total protein profiles are less polymorphic than DNA-based markers the polymorphism in those could be directly associated with fruit quality traits. The aim of this work was to identify associations between polymorphic polypeptides from fruit pericarp at two ripening stages and fruit quality traits evaluated in two segregating populations of tomato. A cross between a normal ripening cultivar of Solanum lycopersicum (C, Caimanta) and a genotype carrying the nor (non ripening) gene (N) as well as a cross between Caimanta and a cherry type tomato of S. lycopersicum var. cerasiforme (Ce) showed genetic variance for several fruit quality traits such as fruit weight, shape, solids soluble content, acidity, color and fruit shelf life. The quantitative variations observed at phenotypic level had correspondence with the polymorphism detected in the protein profiles. Indeed, the polymophic polypeptides associated with quality fruit traits and fruit shelf life would be useful to assist tomato breeding programs as protein molecular markers.     

Spatio-temporal impacts of roads on the persistence of populations: analytic and numerical approaches

Roads can have drastic impacts on wildlife populations. Although there is wide recognition of the negative impacts caused by roads and a wealth of practical studies, there is a lack of theoretical work that can be used to predict the impact of road networks or to implement mitigation measures. Here, using Skellam’s diffusion model, we develop analytic and numerical approaches to analyze the impact of road networks on the survival of populations. Our models show that the viability of a population is determined not only by road density but also by the size and shape of patches. Accordingly, we studied the minimum size of a patch to sustain a population with given diffusion and growth parameters. We provide simple formulas to estimate the minimum patch size, and illustrate the importance of shape with square and rectangular patches. Our models also allow the estimation of time to extinction after road construction for a population in a patch smaller than that of the minimum size. Finally, using numerical computations we illustrate how the spatial arrangement of fences strongly affects both the equilibrium density and the spatial distribution of populations, and that not all fence layouts are equally effective. We anticipate that our methods provide a tool to assess the impact of geometrical features of road networks on wildlife and that they can be used to design mitigation measures to prevent the decline and extinction of populations in an anthropogenically disturbed landscape.     

Fluorescence detection of aluminum in arbuscular mycorrhizal fungal structures and glomalin using confocal laser scanning microscopy

Arbuscular mycorrhizal spores and glomalin-related soil protein (GRSP) isolated from acid soils were analyzed using confocal laser scanning microscopy (CLSM) for Al detection. Mycorrhizal structures of Glomus intraradices produced under in vitro conditions as well as spores and GRSP from neutral and Cu-polluted soils were used as contrasting criteria. Spores and GRSP from soils with 7 and 70% Al saturation showed autofluorescence which increased especially at the highest soil Al level and when Al³⁺ solution was added. G.intraradices spores showed fluorescence only when exogenous Al³⁺ was added. On the contrary, spores and GRSP from neutral and Cu-polluted soils showed little or no significant fluorescence. This fluorescence shown by fungal structures and GRSP when subjected to high Al (of endogenous or exogenous origin) suggest a high capacity for Al immobilization, which could be an effective way to reduce Al activity and phytotoxicity in acid soils.     

Stem modeling and simulation of conversion of cork oak stems for quality wood products

Recent research is underway to study cork oak (Quercus suber L.) wood potential for production of wood components. A total of 35 stems of young and mature cork oaks were sampled, live sawn into flitches, scanned using VTT’s WoodCIM^®, and the measured data computed by VTT’s PuuPilot software, allowing stem 3D reconstruction. Sawing simulations were run for 0.5 m and 1 m logs and the whole stem. Sawn products were (1) planks, (2) parquet, (3) lamparquet, and (4) external component of multilayer planks. Cork oak stems showed a moderate to small taper (mean 24 mm/m). Curviness varied between straight to significantly crooked stems (mean value 40 mm) Batch yields for the tested products ranged 25–43% and 37–50% for 0.5 m logs of young and mature trees, respectively; for 1 m logs, batch yields ranged 19–41% and 25–54%. When using the whole stem, batch yields were lower, ranging 11–38% and 15–50%. Higher yields were obtained for all log lengths and samples for production of lamparquet, parquet, and multilayer component.     

Long-term response of weed control intensity on Scots pine survival, growth and nutrition on former arable land

The effects of post-planting weed control intensity on the cover and composition of ground vegetation, and on growth, survival and foliar nutrient concentrations of 4-year old bare-rooted Scots pine (Pinus sylvestris L.) seedlings were studied over a 15-year period with experimentation on former arable land. Weed control treatments with terbuthylazine and glyphosate were carried out 1?C3 times during successive years, either as overall or as spot applications. The vegetation cover and the shading effect of vegetation decreased with increasing weed control intensity. The more intensive the weed control was, the better was the afforestation result in terms of tree seedling growth and survival. Overall application repeated three times increased seedling survival by 79 percentage points, and their final height, breast height, diameter, and stand volume after 15 years were increased by 183, 19, 15, and 822%, respectively. Weed control did not affect the foliage nutrient concentrations, except for magnesium. However, increase in weed control intensity was reflected in larger needle size 5 years after planting.