NMR-detected conformational exchange observed in a computationally designed variant of protein Gbeta1

Detailed biophysical characterization of computationally designed proteins has become increasingly important in order to thoroughly understand the properties of these variants compared with wild-type and to apply this knowledge to future designs. The protein dynamics and structural properties of a computationally designed variant (Delta1.5) of the beta1 domain of streptococcal protein G (Gbeta1) were measured using multinuclear NMR methods. Results from relaxation, diffusion and hydrogen exchange experiments indicate that the variant weakly self-associates at NMR concentrations, with evidence for multiple binding sites. Although comparison of fast (ps-ns) timescale motions shows only small differences in dynamics between Delta1.5 and wild-type, results from the measurement of intermediate (mus-ms) timescale motions are very different. Significant backbone conformational exchange has been observed in the variant at positions all along the sequence, whereas the wild-type Gbeta1 shows little evidence for this type of motion. This increased conformational exchange in Delta1.5 has been attributed to core overpacking resulting from the incorporation of two large hydrophobic side chains and the loss of an aromatic T-stacking interaction. These data highlight, in detail, the potential consequences of incorporating major perturbations in the core of a protein and the need to carry out more detailed analyses of the biophysical properties of designed proteins in order to better understand and predict the effects of mutations.     

Long-term impact of acid resin waste deposits on soil quality of forest areas I. Contaminants and abiotic properties

Acid resins are residues characterised by elevated concentrations of hydrocarbons and trace elements, which were produced by mineral oil industries in Central Europe during the first half of the last century. Due to the lack of environmental legislation at that time, these wastes were dumped into excavated ponds in public areas without further protection. In this work, the long-term effects of such resin deposits on soil quality of two forest areas (Bayern, Germany) were assessed. We evaluated the distribution and accumulation of contaminants in the surroundings of the deposits, where the waste was disposed of about 60 years ago. General soil chemical properties such as pH, C, N and P content were also investigated. Chemical analysis of resin waste from the deposits revealed large amounts of potential contaminants such as hydrocarbons (93 g kg(-1)), As (63 mg kg(-1)), Cd (24 mg kg(-1)), Cu (1835 mg kg(-1)), Pb (8100 mg kg(-1)) and Zn (873 mg kg(-1)). Due to the location of the deposits on a hillside and the lack of adequate isolation, contaminants have been released downhill despite the solid nature of the waste. Five zones were investigated in each site: the deposit, three affected zones along the plume of contamination and a control zone. In affected zones, contaminants were 2 to 350 times higher than background levels depending on the site. In many cases, contaminants exceeded the German environmental guidelines for the soil-groundwater path and action levels based on extractable concentrations. Resin contamination yielded larger total C/total N ratios in affected zones, but no clear effect was observed on absolute C, N and P concentrations. In general, no major acidification effect was reported in affected zones.     

Long-term impact of acid resin waste deposits on soil quality of forest areas II. Biological indicators

In this study, we evaluated the effects of two acid resin deposits on the soil microbiota of forest areas by means of biomass, microbial activity-related estimations and simple biological ratios. The determinations carried out included: total DNA yield, basal respiration, intracellular enzyme activities (dehydrogenase and catalase) and extracellular enzyme activities involved in the cycles of C (beta-glucosidase and chitinase), N (protease) and P (acid-phosphatase). The calculated ratios were: total DNA/total N; basal respiration/total DNA; dehydrogenase/total DNA and catalase/total DNA. Total DNA yield was used to estimate soil microbial biomass. Results showed that microbial biomass and activity were severely inhibited in the deposits, whilst resin effects on contaminated zones were variable and site-dependant. Correlation analysis showed no clear effect of contaminants on biomass and activities outside the deposits, but a strong interdependence with natural organic matter related parameters such as total N. In contrast, by using simple ratios we could detect more stressful conditions in terms of organic matter turnover and basal metabolism in contaminated areas compared to their uncontaminated counterparts. These results stress that developed ecosystems such as forests can buffer the effects of pollutants and preserve high functionality via natural attenuation mechanisms, but also that acid resins can be toxic to biological targets negatively affecting soil dynamics. Acid resin deposits can therefore act as contaminant sources adversely altering soil processes and reducing the environmental quality of affected areas despite the solid nature of these wastes.     

Semi-natural grassland continuity, long-term land-use change and plant species richness in an agricultural landscape on Öland, Sweden

The study characterizes historical land-use change and the development of semi-natural grassland habitats, over 274 years, within a mosaic agricultural landscape (22 km²) on the island of Öland (Sweden). We also explore the relationship between previous land-use, habitat continuity and present-day vascular plant species richness in grassland patches. Land-cover maps, based on cadastral maps and aerial photographs, were produced for six time-periods between 1723/1733 and 1994/1997. In 1723/1733, the landscape was dominated by grasslands, with arable land surrounding the villages. The grassland area decreased throughout the study period and grassland patches became progressively more fragmented. Present-day grasslands represent 18% of the grassland area in 1723/1733. The land-use structure of the early 18th century is still evident in the modern landscape. The majority of the present-day grasslands are situated on former common grazing land and have had a continuity of at least 274 years: the remaining grasslands are younger and developed during the 20th century on arable or forested land. The proportion of plant species that depend on grazing and are characteristic of semi-natural grasslands significantly reflects the continuity and previous land-use of grassland sites. The study illustrates the way in which information on historical land-use and habitat continuity can help to explain the structuring of plant assemblages in semi-natural grasslands within the modern landscape.     

提高大坝安全性能的一种方法——溢洪道维修案例分析(英文)

瑞典新的洪水设计及大坝安全指南对全国水库大坝的安全提出了更高的要求。指南对每一条河流情况进行了复核并找到了较好的解决方案,即储水和提高溢洪道泄洪能力相结合的方案。为了符合指南的要求,瑞典全国上下都在实施一个大坝安全计划,对大量的水库大坝提高其安全性能,以满足更高的洪水设计(及其它方面)的要求。技术方案包括大坝加固、溢洪道泄洪能力提升和临时储水设施。Stenkullafor 水库大坝设有一个 2 闸控制的敞开式溢洪道。该水库修改后的设计洪水将比现有溢洪道泄洪能力提高 40%。根据成本效益分析,提高溢洪道泄流能力的主要措施是降低其中一个闸门顶高程,以适应需要提高的泄洪能力。通过水工模型试验,一方面优化溢洪道顶部轮廓,另一方面降低溢洪道顶部改造和新闸门的成本。溢洪道右侧闸门底高程降低 4.3 m,并新安装一扇更高的闸门。现在,大坝的安全性能已得到提高。对水工模型试验和施工时的洪水风险,施工过程及其遇到的困难进行了介绍,并对坝体加固、上游坡新的侵蚀防护、下游消能及下游行洪渠道等工程措施进行了描述。     

Influence of liquid binder dispersion on agglomeration in an intensive mixer

In industrial scale mixer granulation, liquid binder is usually sprayed onto the agitated powder bed by means of a nozzle in order to enhance the agglomeration process. The early stage of this process, where granule nuclei are formed and grow, is not well understood. As it is desirable to model the agglomeration state right from the beginning of the process for the purposes of control and modeling, this nucleation step is therefore an important field of interest.To investigate the influence of binder droplet size on the nucleation stage of the agglomeration process, experiments were carried out with lactose and water in an intensive mixer. Water was sprayed in to the mixer with different nozzles to vary the size of the produced droplets. As a comparison, water was also directly poured into the turning mixer. Samples of the produced granules were taken at specific time intervals and analysed for size and water content. As the experiments were focused on examining short granulation times, the first samples were taken after only half of the water was added.Particle size distribution and liquid distribution in the wet granule samples were analyzed. It was found, that the droplet size of the binder liquid has great influence on agglomerate size and binder distribution at short mixing times, with increasing time, the mechanical stresses acting in the mixer becomes more and more dominating in the process. Preliminary comparisons are also carried out with single drop penetration tests in an attempt to correlate drop size to penetration time and also to produced granule size.In conclusion this paper studies the effect of different drop size conditions and subsequent spray flux on the behaviour of the nucleation and the early stages of the agglomeration process. The context of these findings for agglomeration in an intensive mixer is examined.     

Biosynthesis of the vitamin E compound delta-tocotrienol in recombinant Escherichia coli cells

The biosynthesis of natural products in a fast growing and easy to manipulate heterologous host system, such as Escherichia coli, is of increasing interest in biotechnology. This procedure allows the investigation of complex natural product biosynthesis and facilitates the engineering of pathways. Here we describe the cloning and the heterologous expression of tocochromanol (vitamin E) biosynthesis genes in E. coli. Tocochromanols are synthesized solely in photosynthetic organisms (cyanobacteria, algae, and higher green plants). For recombinant tocochromanol biosynthesis, the genes encoding hydroxyphenylpyruvate dioxygenase (hpd), geranylgeranylpyrophosphate synthase (crtE), geranylgeranylpyrophosphate reductase (ggh), homogentisate phytyltransferase (hpt), and tocopherol-cyclase (cyc) were cloned in a stepwise fashion and expressed in E. coli. Recombinant E. coli cells were cultivated and analyzed for tocochromanol compounds and their biosynthesis precursors. The expression of only hpd from Pseudomonas putida or crtE from Pantoea ananatis resulted in the accumulation of 336 mg L(-1) homogentisate and 84 microg L(-1) geranylgeranylpyrophosphate in E. coli cultures. Simultaneous expression of hpd, crtE, and hpt from Synechocystis sp. under the control of single tac-promoter resulted in the production of methyl-6-geranylgeranyl-benzoquinol (67.9 microg g(-1)). Additional expression of the tocopherol cyclase gene vte1 from Arabidopsis thaliana resulted in the novel formation of a vitamin E compound-delta-tocotrienol (15 microg g(-1))-in E. coli.     

Direct imaging of single-walled carbon nanotubes in cells

The development of single-walled carbon nanotubes for various biomedical applications is an area of great promise. However, the contradictory data on the toxic effects of single-walled carbon nanotubes highlight the need for alternative ways to study their uptake and cytotoxic effects in cells. Single-walled carbon nanotubes have been shown to be acutely toxic in a number of types of cells, but the direct observation of cellular uptake of single-walled carbon nanotubes has not been demonstrated previously due to difficulties in discriminating carbon-based nanotubes from carbon-rich cell structures. Here we use transmission electron microscopy and confocal microscopy to image the translocation of single-walled carbon nanotubes into cells in both stained and unstained human cells. The nanotubes were seen to enter the cytoplasm and localize within the cell nucleus, causing cell mortality in a dose-dependent manner.     

Scanning tunnelling spectroscopy on arrays of CdSe quantum dots: response of wave functions to local electric fields

We use scanning tunnelling microscopy (STM) to controllably contact individual CdSe quantum dots (QDs) in a multilayer array to study electrical contacts to a model QD solid. The probability of electron injection into the QD array depends strongly on the symmetry of the QD wave functions and their response to the local electric field. Quantitative spectroscopy of the QD energy levels is possible if the potential distribution in the STM tip-QD array-substrate system is taken into account.     

Industrial Practice of BiPCI Process of Pulverized Coal Injection for Blast Furnace Ironmaking at SSAB

BiPCI technology of pulverized coal injection for blast furnace ironmaking was applied to No.2 Blast Furnace of SSAB Oxelösund in Sweden, and notable effectiveness of this BiPCI practice at SSAB has been achieved. The results show that if the tuyere injection rate of pulverized coal is kept nearly unchanged, (1) BiPCI can increase the overall coal rate and decrease the coke rate. When the second injection rate of pulverized coal attains 5 kg/tHM, the coke rate could be reduced by 5 kg/tHM; (2) BiPCI can increase the burden permeability, decrease the pressure drop of the furnace, and produce proper gas flow distribution, which is favourable to keep smooth running of the blast furnace and decrease the total reducing agent rate (RAR). During the test, the RAR showed a decrease by 2.8 kg/tHM (corrected RAR by 1.45 kg/tHM); (3) The pulverized coal through the second injection can be effectively used to protect the coke from fast degradation and improve the coke strength in the blast furnace, which is favourable to lower the high requirement on the coke quality under high coal rate operation.