Circular single-stranded DNA as a programmable vector for gene regulation in cell-free protein expression systems.

Cell-free protein expression (CFE) systems have emerged as a critical platform for synthetic biology research. The vectors for protein expression in CFE systems mainly rely on double-stranded DNA and single-stranded RNA for transcription and translation processing. Here, we introduce a programmable vector - circular single-stranded DNA (CssDNA), which is shown to be processed by DNA and RNA polymerases for gene expression in a yeast-based CFE system. CssDNA is already widely employed in DNA nanotechnology due to its addressability and programmability. To apply above methods in the context of synthetic biology, CssDNA can not only be engineered for gene regulation via the different pathways of sense CssDNA and antisense CssDNA, but also be constructed into several gene regulatory logic gates in CFE systems. Our findings advance the understanding of how CssDNA can be utilized in gene expression and gene regulation, and thus enrich the synthetic biology toolbox.

Lignocellulosic ethanol and butanol production by Saccharomyces cerevisiae and Clostridium beijerinckii co-culture using non-detoxified corn stover hydrolysate.

Considering global economic and environmental -benefits, green renewable biofuels such as ethanol and butanol are considered as sustainable alternatives to fossil fuels. Thus, developing a co-culture strategy for ethanol and butanol production by Saccharomyces cerevisiae and Clostridium beijerinckii has emerged as a promising approach for biofuel production from lignocellulosic biomass. This study developed a co-culture of S. cerevisiae and C. beijerinckii for ethanol and butanol production from non-detoxified corn stover hydrolysate. By firstly inoculating 3 % S. cerevisiae and then 7 % C. beijerinckii with 8-10 h time intervals, the optimized co-culture process gave 24.0 g/L ABE (20.8 g/L ethanol and 2.4 g/L butanol), obtaining ABE yield and productivity of 0.421 g/g and 0.55 g/L/h. The demonstrated co-culture strategy made full use of hexose and pentose in hydrolysate and contributed to total yield and efficiency compared to conventional ethanol or ABE fermentation, indicating its great potential for developing economically feasible and sustainable bioalcohols production.

Circular single-stranded DNA as switchable vector for gene expression in mammalian cells.

Synthetic gene networks in mammalian cells are currently limited to either protein-based transcription factors or RNA-based regulators. Here, we demonstrate a regulatory approach based on circular single-stranded DNA (Css DNA), which can be used as an efficient expression vector with switchable activity, enabling gene regulation in mammalian cells. The Css DNA is transformed into its double-stranded form via DNA replication and used as vectors encoding a variety of different proteins in a wide range of cell lines as well as in mice. The rich repository of DNA nanotechnology allows to use sort single-stranded DNA effectors to fold Css DNA into DNA nanostructures of different complexity, leading the gene expression to programmable inhibition and subsequently re-activation via toehold-mediated strand displacement. The regulatory strategy from Css DNA can thus expand the molecular toolbox for the realization of synthetic regulatory networks with potential applications in genetic diagnosis and gene therapy.

Multi-objective optimization method for coil current waveform of transcranial magnetic stimulation.

Transcranial magnetic stimulation (TMS) has been proved to be effective in the treatment of many kinds of mental diseases. However, the clicking noise produced by the pulse current with large amplitude and short duration in the TMS coil may damage the hearing of patients. The heat produced by the high-frequency pulse current in the coil also reduces the efficiency of TMS equipment. A multi-objective waveform optimization method to improve heat and noise problems at the same time is presented. By analyzing the current waveforms of TMS, the relationship between the current and the vibration energy/Joule heating is established. Taking the Joule heating and the vibration energy as the optimization objectives, exceeding the same amount of neuronal membrane potential as the limiting condition, the Pareto fronts of different current models are obtained by applying the multi-objective particle swarm optimization algorithm (MOPSO). Therefore, the corresponding current waveforms are inversely deduced. A ringing suppression cTMS (RS-cTMS) proof-of-principle experimental platform is constructed. The feasibility of the proposed method is validated through experiments. The results show that the optimized current waveforms can greatly reduce the vibration and heating of the coil compared with the conventional full-sine, recified sine and half-sine waveforms, thus reducing the pulse noise and prolonging the using time of the equipment. The optimized diversified waveforms also provide a reference for the diversity of TMS.

Identifying Heterozipper ß-Sheet in Twisted Amyloid Aggregation.

Amyloid peptide (AP) self-assembly is a hierarchical process. However, the mechanistic rule of guiding peptides to organize well-ordered nanostructure in a clear and precise manner remains poorly understood. Herein we explored the molecular insight of AP motif aggregates underlying hierarchical process with helical fibrillar structure by atomic force microscope, cryo-electron microscopy (cryo-EM), and molecular dynamics simulation. AP assembly encompasses well-ordered twisted fibrils with uniform morphology, size, and periodicity. More importantly, a heterozipper ß-sheet was identified in a protofilament of AP assembly determined by cryo-EM with a high resolution of 3.5 Å. Each peptide heterozipper was further composed of two antiparallel ß strands and arranged by an alternative manner in a protofilament. The hydrophobic core and hydrophilic area in each zipper played the significant role for peptide assembling. This work proposed and verified the rule facilitating the basic building unit to form twisted fibrils and gave the explanation of peptide hierarchical assembling.

Evolution and Taxonomic Significance of Seed Micromorphology in Impatiens (Balsaminaceae).

Impatiens is one of the most species-rich genera of angiosperms, with more than 1,000 species distributed in Eurasia and Africa. Previous studies have shown that seeds of Impatiens display enormous morphological diversity, and seed micromorphology may contribute to the classification of those species. However, the micromorphological seed coat characteristics of Impatiens seeds have not been systematically studied. In this study, we examined 117 Impatiens seeds from two subgenera and seven sections, and analyzed the seed characters of shape, primary ornamentation, secondary ornamentation, anticlinal cell wall, and periclinal cell wall. We discovered that, according to the different shapes of ornamentation, seed coat can be divided into three types, viz, reticulate type, protrusive type, appendicular type, and that they can be further subdivided into 10 subtypes. In addition, the characteristics of seed coat ornamentation with taxonomic significance in this genus are discussed. We reconstructed the ancestral states of the above seed characters of the Impatiens using the maximum likelihood approach based on the reconstructed phylogenetic framework. The seed character reconstruction showed that the seed shape, primary ornamentation, and anticlinal cell wall could be identified as unambiguous, while other characters were ambiguous in the last common ancestor of Impatiens. Reconstruction of important seed morphological characters showed that the secondary ornamentation possesses high plasticity, and the primary ornamentation has high homology. In addition, we inferred the evolutionary trends of seed ornamentation and found that the seed coat ornamentation of Impatiens experienced a complex evolutionary process from a reticulate type into more complex types. We also discussed the relationship between seed coat diversity vs. environmental adaptation and seed dispersal in Impatiens.

Apoptosis-like bacterial death modulated by photoactive hyperthermia nanomaterials and enhanced wound disinfection application.

Photothermal therapy (PTT) is considered as an efficient therapeutic strategy for wound disinfection. However, there is a dilemma that on the one hand, the high PTT temperature for killing bacteria (>58 °C) could cause serious injury to normal tissue, however, low-temperature results in unsatisfactory treatment efficiency. To settle the issue, we have proposed a novel approach to gently kill bacteria in an apoptosis-like mode via PTT, in which the bacteria can maintain intact membranes but cannot proliferate. This is different from the typical necrosis-like mode of bacterial cell death requiring higher temperatures. We found that PTT prefers to trigger the gradual efflux of Ca2+/Mg2+ ions from the bacterial intracellular content rather than directly destroy the outer membranes, but can cause the dynamic variation of the membrane surface micromorphology. Hence, the microbial viability of E. coli can be dynamically changed from the live state to an apoptosis-like state (45-55 °C), then to apoptosis/necrosis (ca. 58 °C), and finally to necrosis (>61 °C). Based on this strategy, we can kill bacteria through an apoptosis-like mode. Better healing efficacy of mice wounds was achieved at a PTT temperature of 50 °C as compared to that at 58 °C, which sheds light on the wound disinfection and healing applications in clinics with a mild PTT strategy.

Two New Species of Impatiens from China, and Taxonomic Insights into the Longifilamenta Group, Which Is Endemic to China.

Impatiens longshanensis (The LSID for the name Impatiens longshanensis is: 77219154-1) sp. nov. and I. lihengiana (The LSID for the name I. lihengiana is: 77219153-1) sp. nov., from Hunan, China, are described and illustrated here. The molecular phylogenetic study suggests that I. longshanensis and I. lihengiana should be placed in the I. sect. Impatiens. A detailed description, diagnostic characters between the two new species and allied species, pollen and seed morphology, and color photographs are provided. In addition, based on wide sampling, we found that the longifilamenta group, an endemic group to China, whose members have basal lobes of lateral united petals with long filamentous hairs, shows significant morphological variability. In this paper, we discuss the taxonomic significance of morphological characteristics within this group. Based on a literature review and observation of living materials in the field, an updated identification key for this group is also proposed.

Structural conversion of human islet amyloid polypeptide aggregates under an electric field.

Electric fields (EFs) in biological systems are well known, and their presence implies the activity of protein ion channels and pumps in various cells. The aggregation of islet amyloid polypeptides (IAPP) was recently found in human brain tissue, and this was related to the electrical activity of neurons and caused neuronal loss. However, the association between amyloid formation and the electric field is still unknown. Herein a direct method to stimulate the formation of the hIAPP peptide under an EF is reported.

Disassembly of Alzheimer's amyloid fibrils by functional upconversion nanoparticles under near-infrared light irradiation.

Alzheimer's disease (AD) is a common neurodegenerative disease characterized by cognitive and memory function impairment. Studies have shown that the overproduction and further misfolding of amyloid polypeptide (Aß) are the main causes of AD. Therefore, how to reduce Aß species, especially to disassemble the preformed amyloid aggregates, has become the focus of related research. Photodynamic therapy (PDT) using normal photosensitizers eg. porphyrins, is a traditional way of inhibiting amyloid aggregation or degrading the amyloid aggregates, but UV light irradiation presents a side effect for the damage of normal tissue, which limits its medical application. Upconversion nanoparticles (UCNPs) is a luminous material which can be modified by photodynamic agent to form the complex generating reactive oxygen species (ROS) with noninvasive light irradiation. It presents a good advantage in the disassembly of amyloid aggregates via ROS in noninvasive light irradiation. Herein, we prepared ß-NaYF4:Yb/Er@SiO2@RB by combing UCNPs with photosensitizer to disassemble preformed Aß aggregates under near-infrared light. The results displayed upconversion nanomaterials could degrade Aß1-42 fibrils effectively by photo-generating ROS, which demonstrated that the functional UCNPs may have potential applications in the treatment of amyloid diseases in future.

Bioinspired Synthesis of Au Nanostructures Templated from Amyloid ß Peptide Assembly with Enhanced Catalytic Activity.

Peptides have been regarded as useful biomolecule templates to control the synthesis of various inorganic nanomaterials in mild conditions. Inspired by this, the easily self-assembled amyloid ß (Aß) peptide was developed as an alternative template to prepare Au nanostructures for the enhanced catalytic activity, for instance, the reduction of 4-nitrophenol. The presence of Aß peptide assemblies with different structures could direct the nucleation of Au to form different Au nanostructures. Using the Aß25-35 monomers, nanoribbons, and nanofibrils prepared by the self-assembly in phosphate buffered (PB) solution at 0, 3, and 12 h, respectively, as templates could controllably prepare Au nanospheres, nanoribbons, and nanofibers, while the Aß25-35 monomers prepared by the self-assembly in water at 0 h could direct the synthesis of Au nanoflowers. The Aß25-35-templated Au nanostructures had different catalytic activities due to the size and structure effects, which however are significantly enhanced as compared with the template-free Au nanoparticles.

Nanostructural Differentiation and Toxicity of Amyloid-ß25-35 Aggregates Ensue from Distinct Secondary Conformation.

Amyloid nanostructures are originated from protein misfolding and aberrant aggregation, which is associated with the pathogenesis of many types of degenerative diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD) and Huntington's disease. The secondary conformation of peptides is of a fundamental importance for aggregation and toxicity of amyloid peptides. In this work, Aß25-35, a fragment of amyloid ß(1-42) (Aß42), was selected to investigate the correlation between secondary structures and toxicity of amyloid fibrils. Furthermore, each aggregation assemblies show different cell membrane disruption and cytotoxicity. The structural analysis of amyloid aggregates originated from different secondary structure motifs is helpful to understand the mechanism of peptides/cell interactions in the pathogenesis of amyloid diseases.

Removal of cadmium from aqueous solution using low molecular weight chitosan derivative.

A 2(3) factorial design was used to examine the sorption of Cd(2+) onto cross-linked low molecular weight chitosan pyruvic acid derivative. Three factors and two levels of solution pH (A) (6.0 or 10.0), temperature (B) (45 or 70 °C) and Cd(II) concentration (C) (1 or 3 mg/L) were considered. Batch mode system was employed with 0.05 g of the sorbent and 25 mL of Cd(II) solution. The efficiency of cadmium removal during an exposition time of 4h was then evaluated. The factors and their interaction effect on the cadmium removal efficiency followed the order: Cd(II) concentration > solution pH > interaction between solution pH and Cd(II) concentration>interaction between solution pH, temperature and Cd(II) concentration. Langmuir isotherm model was the best isotherm model. The Pseudo second order fitted well the kinetic data. The thermodynamic studies revealed the nature of the cadmium sorption.