Study of accumulation behaviour of tungsten based composite using electron probe micro analyser for the application in bone tissue engineering

In this research, a proto-type study we have conducted, where we have synthesized tungsten based composite materials which are tungsten along with combined oxides of other elements like calcium, scandium, barium, and aluminium in the form of powder with bones powder of mice devised by high energy ball mill and later on fabricating high dense pellets by sintering by spark plasma. The particle sizes of the composite materials are found to be 1–2 µm, as evidenced by the electron microscope, suggesting synthesized materials are of micron size. The quantitative and qualitative analysis of sintered pellets are well confirmed by electron probe micro analyzer (EPMA) and energy dispersive X-ray spectrometer (EDS) which illustrate the greater percentage of tungsten presents in the profound scan areas with other elements of the composite. The absence of pores across the 3D geometry suggesting dense sample, which is quite revealed by the X-ray tomography inspection. The prepared sintered pellets from the tungsten based composites are found to be ≈ 99.5% density with the observation of tungsten to be accumulated uniformly across the scan regions along with focussed hot spots as implied by EPMA. This study paves the way, to examine how the tungsten accumulation and the distribution with the other elements for future understanding in bone tissue engineering application and the in vivo specification of tungsten.     

Planktonic indices in the evaluation of the ecological status and the trophic state of the longest lake in Poland

Due to the intensive mixing polymictic lakes should be homogenous. However, morphometric diversity and high water dynamics contribute to the differentiation of many parameters in various areas of the lakes. This study analyzes both phytoplankton and zooplankton to assess differences in water quality along the north–south axis of the longest lake in Poland. New phytoplankton indicators were applied for determining the lake's ecological status: the Q index based on functional groups and the PMPL (Phytoplankton Metric for Polish Lakes) index based on phytoplankton biomass. TSI_ROT index (Rotifer Trophic State Index), which comprises the percentage of species indicating a high trophic state in the indicatory group and the percentage of bacteriovorus in the Rotifera population, was used for zooplankton analysis.TP content was different at different sites – we observed its gradual increase from the south to the north. Spatial variation of phosphorus did not considerably affect plankton diversity. The phytoplankton was dominated by Oscillatoriales, typical of shallow, well-mixed eutrophic lakes. The ecological status of the lake based on the EQR (Ecological Quality Ratio) was poor or moderate. The zooplankton was dominated by rotifers (at almost all sites), which indicates a eutrophic state of the lake. The values of phytoplankton indices at the studied sites did not differ considerably; the differences resulted more from local conditions such as the contaminant inflow and the macrophyte development than water dynamics.We have demonstrated that in the lake dominated by filamentous Cyanobacteria the ecological status should be determined according to the PMPL index or other indices dependent on the dominant Cyanobacteria species. Since the Q index does not include the functional group S1, the results can lead to the false conclusion that water quality improves with an increased amount of phytoplankton. The high abundance of Cyanobacteria in the lake may have contributed to the poor growth of rotifers.     

The use of ecological integrity indicators within the natural capital index framework: The ecological and economic value of the remnant natural capital of México

In this paper, the ecological integrity hierarchy framework (EIHF) and the natural capital index framework (NCI) are integrated as decision-making tools for evaluating the natural capital of Mexico. Two hierarchy-levels of ecological integrity indicators are used to estimate the quality and quantity of the natural capital, the amount of ecological degradation and ecological sustainability. After human transformation, the extent still considered as “natural” in the country is ∼67%; while the amount of human transformed areas is ∼33%, which gives a total estimate of NCI = 0.334; i.e., only ∼33.4% of the national capital remains available, while ∼33% is ecologically degraded. Furthermore, the critical natural capital; i.e., the legacy for future generations that remains in the country is only ∼12%. The total estimated value of the current natural capital in Mexico is ∼$457.1 billion/yr, which is ∼435 times greater than the national GDP ($1.051 billion in 2010). The cost of maintaining the degradation of the natural capital is ∼$144.6 billion/yr (∼138 times greater than national GDP in 2010). The potential value of the natural capital after restoration would be ∼$602 billion/yr. Valuing the natural capital can be helpful for strategic environmental evaluations and useful for spatial decision support systems that evaluate natural capital as a decision-making tool.     

Rapid production of a chimeric antibody-antigen fusion protein based on 2A-peptide cleavage and green fluorescent protein expression in CHO cells

To enable large-scale antibody production, the creation of a stable, high producer cell line is essential. This process often takes longer than 6 months using standard limited dilution techniques and is very labor intensive. The use of a tri-cistronic vector expressing green fluorescent protein (GFP) and both antibody chains, separated by a GT2A peptide sequence, allows expression of all proteins under a single promotor in equimolar ratios. By combining the advantages of 2A peptide cleavage and single cell sorting, a chimeric antibody-antigen fusion protein that contained the variable domains of mouse IgG with a porcine IgA constant domain fused to the FedF antigen could be produced in CHO-K1 cells. After transfection, a strong correlation was found between antibody production and GFP expression (r = 0.69) using image analysis of formed monolayer patches. This enables the rapid selection of GFP-positive clones using automated image analysis for the selection of high producer clones. This vector design allowed the rapid selection of high producer clones within a time-frame of 4 weeks after transfection. The highest producing clone had a specific antibody productivity of 2.32 pg/cell/day. Concentrations of 34 mg/L were obtained using shake-flask batch culture. The produced recombinant antibody showed stable expression, binding and minimal degradation. In the future, this antibody will be assessed for its effectiveness as an oral vaccine antigen.     

Tree-rings,a key ecological indicator of environment and climate change

Due to wide spatial distribution, high annual resolution, calendar-exact dating, and high climate sensitivity, tree-rings play an important role in reconstructing past environment and climate change over the past millennium at regional, hemispheric or even global scales, so tree-rings can help us to better understand climate behaviour and its mechanisms in the past and then predict variation trends for the future. In this paper, we will review latest advances in tree-ring-based climate reconstructions in China and their applications in modelling past local/regional climate change, capturing historical climatic extreme events, as well as analyzing their link to large-scale climate patterns.     

Ecological and environmental effects of land use change in rapid urbanization: The case of hangzhou,China

Land use changes sharply under rapid urbanization, yet its ecological and environmental effects are often neglected in land use decisions. Using the case of Hangzhou, China, we analyze the ecological and environmental effects of land use changes, including ecosystem services value (ESV) and carbon emissions, based on Landsat TM images from 1995, 2000, 2005, 2010, and 2014. We found significant ecological and environmental effects of land use changes under rapid urbanization. The value of ecosystem services in Hangzhou decreased from 546.7 million USD in 1995 to 448.97 million USD in 2014, and the ratio of ESV to GDP decreased from 5.8% to 0.6%. The net carbon emissions associated with land use changes increased from 4.26 million tons in 1995 to 15.10 million tons in 2014, mainly due to the increase of built-up land carbon emissions and the decrease of forest land carbon sink. The ESV is unevenly distributed spatially and low ESV spread from the central to the peripheral area. We use scenario analysis to illustrate that economic growth and environmental protection could be coordinated by bringing ecological and environmental effects into land use decisions.     

Probing binding mechanism of interleukin-6 and olokizumab: in silico design of potential lead antibodies for autoimmune and inflammatory diseases

Computer-aided antibody engineering has been successful in the design of new biologics for disease diagnosis and therapeutic interventions. Interleukin-6 (IL-6), a well-recognized drug target for various autoimmune and inflammatory diseases such as rheumatoid arthritis, multiple sclerosis, and psoriasis, was investigated in silico to design potential lead antibodies. Here, crystal structure of IL-6 along with monoclonal antibody olokizumab was explored to predict antigen–antibody (Ag???Ab)-interacting residues using DiscoTope, Paratome, and PyMOL. Tyr56, Tyr103 in heavy chain and Gly30, Ile31 in light chain of olokizumab were mutated with residues Ser, Thr, Tyr, Trp, and Phe. A set of 899 mutant macromolecules were designed, and binding affinity of these macromolecules to IL-6 was evaluated through Ag???Ab docking (ZDOCK, ClusPro, and Rosetta server), binding free-energy calculations using Molecular Mechanics/Poisson Boltzman Surface Area (MM/PBSA) method, and interaction energy estimation. In comparison to olokizumab, eight newly designed theoretical antibodies demonstrated better result in all assessments. Therefore, these newly designed macromolecules were proposed as potential lead antibodies to serve as a therapeutics option for IL-6-mediated diseases.     

The dynamics of experimental arable weed communities under different management practices

Abstract. Weed communities, comprising 12 introduced species at constant starting densities and three species already present in the seed bank, were followed through three years of continuous winter wheat. The wheat and weeds were subjected to two treatments in a split-plot factorial design, organic contrasting with conventional fertilizer, and ploughing plus hand-roguing contrasting with minimum tillage plus herbicide. The minimum tilled plots developed in a uniform manner, and became dominated by very high densities of Anisantha sterilis. Agrostemma githago and Galium aparine also persisted in these plots at lower densities. The ploughed plots had a lower total density but a greater range of species. Stellaria media, Veronica persica and Avena fatua were the most common; other species occurred at lower densities. The major effect of fertilizer treatment was a greater initial increase by G. aparine on the organic, minimum tilled plots compared with the conventionally fertilized, minimum tilled plots. Species associated with minimum tillage were annuals with either no or a short term seed bank and autumn germination and rather predictable dynamics, whereas species that did well under ploughing were either spring germinating or had a persistent seed bank, implying greater annual variation in population size associated with weather conditions. There seemed no clear way to distinguish between those species which were abundant on the ploughed plots and those which were scarce under all conditions using readily accessible data.     

Enhancing plasmid transformation efficiency and enabling CRISPR-Cas9/Cpf1-based genome editing in Clostridium tyrobutyricum

Clostridium tyrobutyricum ATCC 25755 is known as a natural hyper-butyrate producer with great potentials as an excellent platform to be engineered for valuable biochemical production from renewable resources. However, limited transformation efficiency and the lack of genetic manipulation tools have hampered the broader applications of this micro-organism. In this study, the effects of Type I restriction-modification system and native plasmid on conjugation efficiency of C. tyrobutyricum were investigated through gene deletion. The deletion of Type I restriction endonuclease resulted in a 3.7-fold increase in conjugation efficiency, while the additional elimination of the native plasmid further enhanced conjugation efficiency to 6.05 ± 0.75 × 10³ CFU/ml-donor, which was 15.3-fold higher than the wild-type strain. Fermentation results indicated that the deletion of those two genetic elements did not significantly influence the end-products production in the resultant mutant ΔRMIΔNP. Thanks to the increased conjugation efficiency, the CRISPR-Cas9/Cpf1 systems, which previously could not be implemented in C. tyrobutyricum, were successfully employed for genome editing in ΔRMIΔNP with an efficiency of 12.5–25%. Altogether, approaches we developed herein offer valuable guidance for establishing efficient DNA transformation methods in nonmodel micro-organisms. The ΔRMIΔNP mutant can serve as a great chassis to be engineered for diverse valuable biofuel and biochemical production.