Single reconstituted neuronal SNARE complexes zipper in three distinct stages

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins drive membrane fusion by assembling into a four-helix bundle in a zippering process. Here, we used optical tweezers to observe in a cell-free reconstitution experiment in real time a long-sought SNARE assembly intermediate in which only the membrane-distal amino-terminal half of the bundle is assembled. Our findings support the zippering hypothesis, but suggest that zippering proceeds through three sequential binary switches, not continuously, in the amino- and carboxyl-terminal halves of the bundle and the linker domain. The half-zippered intermediate was stabilized by externally applied force that mimicked the repulsion between apposed membranes being forced to fuse. This intermediate then rapidly and forcefully zippered, delivering free energy of 36 k(B)T (where k(B) is Boltzmann's constant and T is temperature) to mediate fusion.     

Nitrex: The timing of response of coniferous forest ecosystems to experimentally-changed nitrogen deposition

In large regions of Europe and eastern North America atmospheric deposition of inorganic nitrogen (N) compounds has greatly increased the natural external supply to forest ecosystems. This leads to N saturation, in which availability of inorganic N is in excess of biological demand and the ecosystem is unable to retain all incoming N. The large-scale experiments of the NITREX project (NITRogen saturation EXperiments) are designed to provide information regarding the patterns and rates of responses of coniferous forest ecosystems to increases in N deposition and the reversibility and recovery of impacted ecosystems following reductions in N deposition.The timing of ecosystem response generally followed a hypothesized cascade of response. In all sites N outputs have responded markedly but to very different degrees within the first three years of treatment. Within this time significant effects on soil processes and on vegetation have only been detected at two sites. This delayed response is explained by the large capacity of the soil system to buffer the increased N supply by microbial immobilization and adsorption. We believe that this concept provides a framework for the evaluation and prediction of the ecosystem response to environmental change.     

Comparison of major and trace element concentrations in Danish greenhouse tomatoes (Lycopersicon esculentum Cv. Aromata F1) cultivated in different substrates

The concentration of major and trace elements was determined for tomato (Lycopersicon esculentumcv. Aromata F1) fruits grown in three different substrate systems. The systems were soil and rockwool irrigated with a normal nutrient solution and rockwool irrigated with a nutrient solution with elevated electrical conductivity (EC). At three harvest times, tomato fruits were analyzed for Ca, Cu, Fe, K, Mg, Mn, Na, P, S, Sr, and Zn by ICP-AES and for Cd, Cr, Mo, Ni, Pb, Sn, and V by HR-ICPMS. The concentrations of Ca, Cd, Fe, Mn, Mo, Na, Ni, Sr, and Zn were significantly different (p < 0.05) for tomato fruits grown on the different substrates. Between the harvest times different levels (p < 0.05) were shown for Ca, Cd, Fe, Mn Na, Ni, Sr, Zn Cu, K, Mg, P, Sn, and V. The concentration of Cd was >15 times higher and the concentration of Ca was 50-115% higher in soil-grown fruits than in rockwool-grown fruits. Principal component analysis applied on each harvest split the data into two groups. One group includes soil-grown fruits, and the other group includes rockwool-grown fruits with the two different nutrient solutions.     

Cryopreservation of European Mouflon (Ovis Gmelini Musimon) Semen During the non-Breeding Season is Enhanced by the Use of Trehalose

The influence of trehalose on European mouflon spermatozoa cryopreservation during the non-breeding season was tested. Semen was frozen in two different extenders: (a) recommended Tris-based ram extender (CTR); (b) CTR extender supplemented with trehalose 0.147 mm (TRH). Sperm viability and acrosome integrity were assessed using propidium iodide and fluorescein isothiocynate labelled Pisum Sativum agglutinin. Trehalose significantly enhanced sperm viability after thawing compared with CTR extender (62.7% vs 51.8%; p < 0.05), whereas no differences were observed on acrosome integrity (42.9% vs 42.1%). Trehalose influence was also evidenced in the in vitro fertility test performed with sheep oocytes matured in vitro. Both fertilization rates (60.9% TRH vs 43.6% CTR; p < 0.05) and cleavage rates (58% TRH vs 39.8% CTR; p < 0.001) were higher for trehalose frozen semen compared with control extender frozen semen. A higher percentage of zygotes resulting from fertilization with trehalose cryopreserved semen presented the first cleavage earlier if compared with the group fertilized with control semen (48.7% vs 31.5%, respectively; p < 0.01). This result was confirmed by embryo kinetic development. Fertilization with trehalose cryopreserved semen leaded to an higher percentage of blastocysts (40.2% vs 27.8% CTR; p < 0.05), and enhanced in particular the number of blastocysts that developed on the day 6th of culture (28.6% vs 17% CTR; p < 0.05). Our data demonstrated that, during mouflon non-breeding season, trehalose extender enhances spermatozoa viability and its in vitro fertilizing capacity, allowing the production of an higher number of blastocysts.     

Assessment of urban forestry research and research needs in Nordic and Baltic countries

A review of research and research needs in urban forestry was carried out in Denmark, Estonia, Finland, Latvia, Norway and Sweden during 2005. A questionnaire addressing post-2000 and ongoing research was sent to 146 researchers and generated 76 completed questionnaires. Universities were found to lead urban forestry research, while municipalities headed funding organisations in terms of number of projects funded. Planning, ecological and management aspects were the most common research themes, but socially oriented research also played an important role. The research needs questionnaire was sent to 192 key research actors (assignors, users and researchers), resulting in 63 completed needs assessments. The research themes of ‘urban forest management’, ‘social and cultural values’ and ‘urban forest and green planning’ were prioritised for future research. Comparison of ongoing research and research needs showed discrepancies, as ongoing research does not always cover the same themes identified as primary research needs. Priorities for future research as identified by the research community respective those assigning and using research also differed. Economic assessment of benefits, for example, scored much higher as a need among researchers than other respondents. In terms of present weaknesses in the research ‘infrastructure’, research actors emphasised lack of funding, fragmentation of research and insufficient critical mass. The region's urban forestry research can be enhanced and made more meaningful by strengthening national and international networking within the research community, across disciplines, as well as between researchers and those commissioning and using research.     

Seedling growth response of two tropical tree species to nitrogen deposition in southern China

Seedling growth response of two tropical tree species (Schima superba and Cryptocarya concinna) to simulated N deposition was studied during a period of 11 months. One-year-old seedlings were grown in forest soil treated with N as NH₄NO₃ at Control–no N addition, N5–5, N10–10, N15–15, and N30–30 g N m⁻² year⁻¹. The objective was to examine the effects of N addition on seedling growth and compare this effect between the two tropical tree species of different species-N-requirement. Results showed that both species responded significantly to N addition and exhibited positive effect to lower rate of N addition and negative effect to higher rate of N addition on growth parameters (height and stem base diameter, biomass production, and net photosynthetic rate). The highest values were observed in the N10 plots for S. superba and in the N15 plots for C. concinna, but the lowest values were observed in the N30 plots for both species. However, the reduction in the N30 plots was more pronounced for S. superba than for C. concinna relative to the control plots. Our findings suggest that response of seedling growth of tropical tree species to atmospheric N deposition may vary depending on rate of N deposition and species-N-requirement.