Metabolic engineering of yeast for increased production of cyclopropane fatty acids

Biological production of chemicals and fuels using whole cells is an important and growing segment of manufacturing and among the various forms, microorganisms are the most successfully utilized. In particular, yeasts such as Saccharomyces cerevisiae are both widely used production organisms and metabolic models for oleaginous yeasts. Fatty acid-containing lipids are one example of moderate value, highly versatile chemicals produced by yeasts that are used in a broad range of industries for lubrication, cosmetics, fuels and polymers. Production levels of standard fatty acids by yeasts has increased enormously over the past 10 years through the application of metabolic pathway engineering, flux analysis, computational approaches and to a lesser extent, bioprocessing improvements. Combined, these advances have brought yeast-based fatty acid production close to commercial reality. Functionalized fatty acids such as those containing hydroxyl or cyclopropyl groups are more valuable as chemical feedstocks and are an attractive target for yeast production as commercial supply is limited. Cyclopropane fatty acids, possessing a strained 3-membered ring and having a saturated chain, are especially attractive as they have application in cosmetics and specialty lubrication. However, cyclopropyl fatty acids present greater challenges for metabolic engineering as they are not produced naturally by yeast. Please click Additional Files below to see the full abstract

Osthole represses growth of multiple myeloma cells by regulating PI3K/AKT and ERK pathways

Purpose: To identify the effects of osthole on the growth of multiple myeloma cells and determine the probable molecular mechanism of action. Methods: 3-(4, 5-Dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to measure multiple myeloma cell viability. Flow cytometry was used for the evaluation of cell cycle and apoptosis. For protein expression measurement, western blot analysis was employed. Results: The results show that osthole dose- and time-dependently suppressed the viability of multiple myeloma cells (p < 0.01). Osthole also dose-dependently initiated multiple myeloma cell cycle arrest in G0/G1 phase (p < 0.05) and induced multiple myeloma cell apoptosis (p < 0.01). Moreover, western blotting revealed a significant reduction in cyclin D1 (p < 0.01) and induction in p21, cleaved caspase 3 and cleaved poly (ADP-ribose) polymerase (p < 0.01). Furthermore, osthole treatment significantly downregulated the phosphorylation levels of protein kinase B (AKTT308), AKTS473, and extracellular regulated protein kinase 1/2 (ERK1/2). Conclusion: These findings demonstrated that osthole inhibited viability and induced cell cycle arrest and apoptosis of multiple myeloma cells by regulating PI3K/AKT and ERK pathways. Osthole may be considered as a potential anticancer agent for the therapy of multiple myeloma

Engineer RNA-protein nanowires as light-responsive biomaterials

RNA molecules have emerged as increasingly attractive biomaterials with important applications such as RNA interference (RNAi) for cancer treatment and mRNA vaccines against infectious diseases. However, it remains challenging to engineer RNA biomaterials with sophisticated functions such as non-covalent light-switching ability. Herein, light-responsive RNA-protein nanowires are engineered to have such functions. It first demonstrates that the high affinity of RNA aptamer enables the formation of long RNA-protein nanowires through designing a dimeric RNA aptamer and an engineered green fluorescence protein (GFP) that contains two TAT-derived peptides at N- and C- termini. GFP is then replaced with an optogenetic protein pair system, LOV2 (light–oxygen–voltage) protein and its binding partner ZDK (Z subunit of protein A), to confer blue light-controlled photo-switching ability. The light-responsive nanowires are long (>500 nm) in the dark, but small (20–30 nm) when exposed to light. Importantly, the co-assembly of this RNA-protein hybrid biomaterial does not rely on the photochemistry commonly used for light-responsive biomaterials, such as bond formation, cleavage, and isomerization, and is thus reversible. These RNA-protein structures can serve as a new class of light-controlled biocompatible frameworks for incorporating versatile elements such as RNA, DNA, and enzymes

Exogenous glutathione improves high root-zone temperature tolerance by modulating photosynthesis, antioxidant and osmolytes systems in cucumber seedlings

To investigate the physiological responses of plants to high root-zone temperature (HT, 35 °C) stress mitigated by exogenous glutathione (GSH), cucumber (Cucumis sativus L.) seedlings were exposed to HT with or without GSH treatment for 4 days and following with 4 days of recovery. Plant physiological variables, growth, and gene expression related to antioxidant enzymes and Calvin cycle were quantified. The results showed that HT significantly decreased GSH content, the ratio of reduced to oxidized glutathione (GSH/GSSG), chlorophyll content, photosynthesis and related gene expression, shoot height, stem diameter, as well as dry weight. The exogenous GSH treatment clearly lessened the HT stress by increasing the above variables. Meanwhile, HT significantly increased soluble protein content, proline and malondialdehyde (MDA) content as well as O2•− production rate, the gene expression and activities of antioxidant enzymes. The GSH treatment remarkably improved soluble protein content, proline content, antioxidant enzymes activities, and antioxidant enzymes related gene expression, and reduced the MDA content and O2•− production rate compared to no GSH treatment in the HT condition. Our results suggest that exogenous GSH enhances cucumber seedling tolerance of HT stress by modulating the photosynthesis, antioxidant and osmolytes systems to improve physiological adaptation

Model Simulation of Cucumber Yield and Microclimate Analysis in a Semi-closed Greenhouse in China

Adequate greenhouse environmental management is very important for improving resource use efficiency and increasing vegetable yield. The objective of this study was to explore suitable climate and cultivation management for cucumber to achieve high yield and build optimal yield models in semi-closed greenhouses. A fruit cucumber cultivar Deltastar was grown over 4 years in greenhouse and weekly data of yields (mean, highest and lowest) and environmental variables, including total radiation, air temperature, relative humidity, and carbon dioxide (CO2) concentration were collected. Regression analyses were applied to develop the relationships and build best regression models of yields with environmental variables using the first 2 years of data. Data collected in years 3 and 4 were used for model validation. Results showed that total radiation, nutrient, temperature, CO2 concentration, and average nighttime relative humidity had significant correlations with cucumber yields. The best regression models fit the mean, lowest, and highest yields very well with R2 values of 0.67, 0.66, and 0.64, respectively. Total radiation and air temperature had the most significant contributions to the variations of the yields. Our results of this study provide useful information for improving greenhouse climate management and yield forecast in semi-closed greenhouses

New Superhard Carbon Phases Between Graphite and Diamond

Two new carbon allotropes (H-carbon and S-carbon) are proposed, as possible candidates for the intermediate superhard phases between graphite and diamond obtained in the process of cold compressing graphite, based on the results of first-principles calculations. Both H-carbon and S-carbon are more stable than previously proposed M-carbon and W-carbon and their bulk modulus are comparable to that of diamond. H-carbon is an indirect-band-gap semiconductor with a gap of 4.459 eV and S-carbon is a direct-band-gap semiconductor with a gap of 4.343 eV. The transition pressure from cold compressing graphite is 10.08 GPa and 5.93 Gpa for H-carbon and S-carbon, respectively, which is in consistent with the recent experimental report.Comment: 5pages,4figures,submitted to Phys.Rev.Lett on 18Jan12, transfer to Phys.Rev.B on 25Mar12; Solid State Communications(2012), http://dx.doi.org/10.1016/j.ssc.2012.05.02

Seismic Vulnerability Evaluation of a Three-Span Continuous Beam Railway Bridge

In order to evaluate the seismic vulnerability of a railway bridge, a nonlinear finite element model of typical three-span continuous beam bridge on the Sichuan-Tibet railway in China was built. It further aimed at performing a probabilistic seismic demand analysis based on the seismic performance of the above-mentioned bridge. Firstly, the uncertainties of bridge parameters were analyzed while a set of finite element model samples were formulated with Latin hypercube sampling method. Secondly, under Wenchuan earthquake ground motions, an incremental dynamic method (IDA) analysis was performed, and the seismic peak responses of bridge components were recorded. Thirdly, the probabilistic seismic demand model for the bridge principal components under the prerequisite of two different kinds of bearing, with and without seismic isolation, was generated. Finally, comparison was drawn to further ascertain the effect of two different kinds of bearings on the fragility components. Based on the reliability theory, results were presented concerning the seismic fragility curves

Parameter optimization of damper-friction isolation systems using concave friction distribution

The random friction distribution is an objective phenomenon and always leads to great uncertainty in seismic responses of damper-friction isolation systems. As to optimize the seismic performance, this paper artificially made the friction distribution on a contact surface to be concave. When the damper-friction isolation system was subjected to different ground motions, more regular responses, due to the concave friction distribution, were obtained comparing with the responses of the random friction distribution. The concave friction distribution is always conducive to reducing the structural relative displacement and insignificantly increasing the structural acceleration. An optimization design example of an isolation building was carried out. And the results showed that as for a structure only being sensitive to acceleration or force, significant concave friction distribution and little damping constant would be the best design combination. It resulted in a much less acceleration and an acceptable relative displacement on the structure

Effects of Heat Shock on Photosynthetic Properties, Antioxidant Enzyme Activity, and Downy Mildew of Cucumber (Cucumis sativus L.)

Heat shock is considered an abiotic stress for plant growth, but the effects of heat shock on physiological responses of cucumber plant leaves with and without downy mildew disease are still not clear. In this study, cucumber seedlings were exposed to heat shock in greenhouses, and the responses of photosynthetic properties, carbohydrate metabolism, antioxidant enzyme activity, osmolytes, and disease severity index of leaves with or without the downy mildew disease were measured. Results showed that heat shock significantly decreased the net photosynthetic rate, actual photochemical efficiency, photochemical quenching coefficient, and starch content. Heat shock caused an increase in the stomatal conductance, transpiration rate, antioxidant enzyme activities, total soluble sugar content, sucrose content, soluble protein content and proline content for both healthy leaves and downy mildew infected leaves. These results demonstrate that heat shock activated the transpiration pathway to protect the photosystem from damage due to excess energy in cucumber leaves. Potential resistance mechanisms of plants exposed to heat stress may involve higher osmotic regulation capacity related to an increase of total accumulations of soluble sugar, proline and soluble protein, as well as higher antioxidant enzymes activity in stressed leaves. Heat shock reduced downy mildew disease severity index by more than 50%, and clearly alleviated downy mildew development in the greenhouses. These findings indicate that cucumber may have a complex physiological change to resist short-term heat shock, and suppress the development of the downy mildew disease