Editing modifies the GABA(A) receptor subunit alpha3

Adenosine to inosine (A-to-I) pre-mRNA editing by the ADAR enzyme family has the potential to increase the variety of the proteome. This editing by adenosine deamination is essential in mammals for a functional brain. To detect novel substrates for A-to-I editing we have used an experimental method to find selectively edited sites and combined it with bioinformatic techniques that find stem-loop structures suitable for editing. We present here the first verified editing candidate detected by this screening procedure. We show that Gabra-3, which codes for the alpha3 subunit of the GABA(A) receptor, is a substrate for editing by both ADAR1 and ADAR2. Editing of the Gabra-3 mRNA recodes an isoleucine to a methionine. The extent of editing is low at birth but increases with age, reaching close to 100% in the adult brain. We therefore propose that editing of the Gabra-3 mRNA is important for normal brain development.     

Effects of an exotic plant and habitat disturbance on pollinator visitation and reproduction in a boreal forest herb

The invasion of exotic species into natural habitats is considered to be a major threat to biodiversity, and many studies have examined how exotic plants directly affect native plant species through competitive interactions for abiotic resources. However, although exotics can have potentially great ecological and evolutionary consequences, very few researchers have studied the effect of exotics on the interactions between plants and their mutualistic partners, such as pollinators, and none have reported on such impacts in logged and undisturbed boreal forest ecosystems. Here we show how experimental introductions of an exotic plant species (Phacelia tanacetifolia Bentham) affect pollinator visitation and female reproductive success of a native plant (Melampyrum pratense L.) in recently disturbed (i.e., logged) and in undisturbed boreal forest habitats. The presence of Phacelia significantly increased the number of bumble bees entering plots in both habitat types. However, the exotic species had a strong negative impact on the visitation rate to the native species in both habitat types. Despite this negative impact on pollinator visitation, the exotic had no effect on female reproductive success of the native species in any habitat. Our results show that seed production may be more robust than pollinator visitation to exotic invasion, irrespective of habitat disturbance history.     

Glycoproteins of Axonal Transport: Affinity Chromatography on Fucose-Specific Lectins

Rapidly transported fucose-labeled glycoproteins from axons of rabbit retinal ganglion cells were solubilized with nonionic detergents. The solubilized components were subjected to affinity chromatography on three different fucose-specific lectins. A recently characterized fucose-specific lectin from Aleuria aurantia bound reversibly approximately 60% of the applied protein-bound radioactivity. The lectins from Lotus tetragonolobus and Ulex europaeus bound are very small proportions of the labeled rapidly transported glycoproteins.     

Phenotypic responses to light,water, and nutrient conditions in the allopolyploid Arabidopsis suecica and its parent species A. thaliana and A. arenosa: Does the allopolyploid outrange its parents?

Polyploid species possess more than two sets of chromosomes and may show high gene redundancy, hybrid vigor, and masking of deleterious alleles compared to their parent species. Following this, it is hypothesized that this makes them better at adapting to novel environments than their parent species, possibly due to phenotypic plasticity. The allopolyploid Arabidopsis suecica and its parent species A. arenosa and A. thaliana were chosen as a model system to investigate relationships between phenotypic plasticity, fitness, and genetic variation. Particularly, we test if A. suecica is more plastic, show higher genetic diversity, and/or have higher fitness than its parent species. Wild Norwegian populations of each species were analyzed for phenotypic responses to differences in availability of nutrient, water, and light, while genetic diversity was assessed through analysis of AFLP markers. Arabidopsis arenosa showed a higher level of phenotypic plasticity and higher levels of genetic diversity than the two other species, probably related to its outbreeding reproduction strategy. Furthermore, a general positive relationship between genetic diversity and phenotypic plasticity was found. Low genetic diversity was found in the inbreeding A. thaliana. Geographic spacing of populations might explain the clear genetic structure in A. arenosa, while the lack of structure in A. suecica could be due to coherent populations. Fitness measured as allocation of resources to reproduction, pointed toward A. arenosa having lower fitness under poor environmental conditions. Arabidopsis suecica, on the other hand, showed tendencies toward keeping up fitness under different environmental conditions.     

Meristem activity and biomass production as response mechanisms in two forest herbs

Actaea spicata and Geranium sylvaticum are deciduous rosette herbs, frequently coexisting in nutrient-rich forests of boreal Sweden. A. spicata is restricted to this habitat whereas G. sylvaticum has a wider ecological amplitude. To explore the different distributions of these species we studied the growth mechanisms by which they responded to nutrient and light supply. Plants raised from seeds were grown under four combinations of light and nutrient supplies and growth was followed for 20 weeks. Growth responses were measured as biomass accumulation and meristem activity, i.e. leaf development by shoot meristems. Biomass accumulation and meristem activity were affected by nutrient and light treatments. However, A. spicata responded more to nutrients, whereas G. sylvaticum responded more to light, and low resource supply shortened the time during which shoot meristems of A. spicata developed leaves without a corresponding effect on biomass accumulation. In G. sylvaticum, the periods of leaf development and biomass accumulation were the same, but a relationship between high development rates and peak allocation of biomass to shoot meristems indicated a storage function of these during leaf development. We conclude that the inflexible growth of A. spicata, mainly affected by nutrient supply, makes it less competitive in open habitats and restricts it to shaded, nutrient-rich habitats. G. sylvaticum should be more successful in habitats subjected to unpredictable disturbances, due to a flexible growth morphology and a dynamic function of meristems that buffers against variation in nutrient supply and facilitates positive light responses. Meristematic data should be included in interspecific comparisons on growth responses to different resource supplies. Unless species have the same growth mechanism, it is not satisfactory to study biomass-related traits alone, as internal constraints concerning plant development might confuse the interpretation.     

Screening for transient biological interactions as applied to albumin ligands: a new concept for drug discovery

The notion that many biological interactions are based on transient binding (dissociation constants (K(d)) in the range of 10-0.01 mM) is familiar, yet the implications for biological sciences have been realized only recently. An important area of biological sciences is drug design, where the traditional "lock and key" view of binding has prevailed and drug candidates are usually selected on their merits as being tight binders. However, the rationale that transient interactions are of importance for drug discovery is slowly gaining acceptance. These interactions may relate not only to the desired target interaction but also to unwanted interactions creating, for example, toxicity problems. Here we demonstrate, in a high-throughput screening format, affinity selection of weak binders to a model target of albumin by zonal retardation chromatography. It is perceived that this approach can define the "transient drug" as a complement to current drug discovery procedures.     

Factors associated with inflammation markers, a cross-sectional analysis

Epidemiological studies have reported associations between circulating inflammation markers and risk of chronic diseases. It is of interest to examine whether risk factors for these diseases are associated with inflammation. We conducted a cross-sectional analysis to evaluate whether reproductive and lifestyle factors and circulating vitamin D were associated with inflammation markers, including C-reactive protein, cytokines (IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p40, IL-12p70, IL-13, TNFα), and cytokine modulators (IL-1RA, sIL-1RII, sIL-2Ra, sIL-4R, sIL-6R, sTNF-R1/R2), among 616 healthy women. We confirmed associations of several inflammation markers with age and BMI. We also observed significantly higher levels of certain inflammation markers in postmenopausal vs. premenopausal women (TNFα, sIL-1RII, sIL-2Ra), with increasing parity (IL-12p40), and with higher circulating 25(OH) vitamin D (IL-13) and lower levels among current users of non-steroidal anti-inflammatory drugs (NSAIDs) (IL-1β, IL-2, IL-10, IL-12p70, and IL-12p40), current smokers (IL-4, IL-13, IL-12p40), and women with a family history of breast or ovarian cancer (IL-4, IL-10, IL-13). Our findings suggest that risk factors for chronic diseases (age, BMI, menopausal status, parity, NSAID use, family history of breast and ovarian cancer, and smoking) are associated with inflammation markers in healthy women.     

Steroid hydroxylation using immobilized spores of Curvularia lunata germinated in situ

Summary Spores of Curvularia lunata were immobilized in polyacrylamide granules and in calcium alginate beads (2–3 mm in diam.). Germination of the spores, initiated by the addition of nutrients, resulted in an even distribution of mycelium throughout the beads after 48 h. Such beads were used for the conversion of cortexolone to cortisol by steroid-11-hydroxylation. In order to improve the steroid transforming ability several parameters were studied. It was found that preparations based on calcium alginate gave the best results.The possible merits of immobilizing spores rather than vegetative cells, followed by in situ germination are discussed also for other microorganisms and immobilization processes.     

Weak affinity chromatography of small saccharides with immobilised wheat germ agglutinin and its application to monitoring of carbohydrate transferase activity

In this work we have evaluated the potential to use wheat germ agglutinin(WGA) for weak affinity chromatography (WAC) of N-acetyl derivatives ofmono-, di-, tri- and tetrasaccharides. WGA was used as a ligand in a highperformance liquid affinity chromatography (HPLAC) system. Isocraticaffinity chromatography was conducted where similar N-acetyl saccharideswere separated according to their binding strength to WGA. Affinities areweak and lie typically in the mM range. For example, for3sialyllactose, the dissociation constant (K_d) wasfound to be 2.4 mM at 8°C. It was interesting to note that theWGA–HPLC column can distinguish between the anomeric forms ofN-acetylglucosamine. Weak affinity chromatography with immobilised WGA wasused in an enzyme assay to detect the activity of GlcNAc-transferases.     

Nutrient and Carbon Additions to the Microbial Soil Community and its Impact on Tree Seedlings in a Boreal Spruce Forest

We have added glucose and nutrients to manipulate soil microbial activity and nutrient availability in a boreal spruce forest to study the performance of birch and spruce seedlings in relation to the soil microbial community. The proportion of aboveground biomass in the seedlings was largest in plots amended with extra nutrients, while ectomycorrhizal (ECM) colonisation was low in these plots. ECM appeared beneficial for growth of both species, but only at low levels of colonisation (<25% ECM colonised root-tips). The soil microbial biomass, as determined by total PLFA, was largest in plots treated with glucose and there was a significant negative relationship between birch seedling size and levels of total PLFA in soil. This could be taken to suggest that poor seedling growth was due to nutrient limitation caused by microbial assimilation. However, the treatment response of the birch seedlings was generally weak, and spruce often showed no response at all to the addition of nutrients and glucose. The most consistent parameter for the variation in plant performance, as well as for the microbial soil community, was the block-effect. This suggests a strong spatial structure in the soil microbial community, and that this structure was robust with respect to our treatments even though they continued over a 3-year period.