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[Methods in Molecular Biology] Plant Long Non-Coding RNAs Volume 1933 (Methods and Protocols) || Trimolecular Fluorescence Complementation (TriFC) Assay for Visualization of RNA-Protein Interaction in Plants
摘要: RNA-protein interactions play important roles in various eukaryotic biological processes. Molecular imaging of subcellular localization of RNA-protein complexes in plants is critical for understanding these interactions. However, methods to image RNA-protein interactions in living plants have not yet been developed until now. Recently, we have developed a trimolecular fluorescence complementation (TriFC) system for in vivo visualization of RNA-protein interaction by transient expression in tobacco leaves. In this method, we combined conventional bimolecular fluorescence complementation (BiFC) system with the MS2 system (phage MS2 coat protein [MCP] and its binding RNA sequence [MS2 sequence]) to tag lncRNA. Target RNA is tagged with 6xMS2, and MCP and RNA-binding protein are fused with YFP fragments. DNA constructs encoding such fusion RNA and proteins are infiltrated into tobacco leaves with Agrobacterium suspensions. RNA-protein interaction in vivo is observed by confocal microscopy.
关键词: RNA-protein interaction,Long noncoding RNA,TriFC,Tobacco transient expression,In vivo visualization
更新于2025-11-21 11:08:12
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Particulate matter emissions of less harmful-looking super-slim size cigarettes appealing to women: a laser spectrometric analysis of second-hand smoke
摘要: Slim tobacco products shall appear by their look less harmful. In 2013, the European Union ministers discussed to ban them. However, only a ban on small package sizes was realized. To add more data for exposure risk assessment of slim tobacco products, the particulate matter (PM) amount in second-hand smoke (SHS) of super-slim size cigarettes compared with a king size brand was investigated. PM amount of four super-slim size cigarette types of the brand Couture was analysed in comparison with the king-size reference cigarette. Therefore, SHS was produced in an enclosed space with a volume of 2.88 m3 by an automatic environmental tobacco smoke emitter. PM size fractions PM10, PM2.5 and PM1 were measured in real time using a laser aerosol spectrometer. SHS of Couture Gold contained about 36% and Couture Purple about 28% more PM than the reference cigarette. In contrast, Couture Green emitted about 37% and Couture Silver about 53% less PM than the reference cigarette. Depending on the brand, the PM2.5 mean concentrations increased up to 1538 μg/m3. This exceeds the 24-h mean concentration of at most 25 μg/m3 according to the WHO Air quality guidelines about 62-fold. Smoking in enclosed rooms leads to a massive increase of PM. The PM pollution by slim-size tobacco products are substantial and sometimes higher than by king size tobacco products. Therefore, SHS exposure from slim-size tobacco products is not less harmful to health. Decision makers should take this aspect in consideration.
关键词: Second-hand smoke,Super-slim size cigarettes,Slim-size cigarettes,Particulate matter,Environmental tobacco smoke
更新于2025-09-12 10:27:22
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Economic Impact of Substituting Solar Photovoltaic Electric Production for Tobacco Farming
摘要: Solar photovoltaic (PV) technology represents a promising method to prevent dangerous global climate change, however full solar penetration demands substantial surface areas, possibly encroaching on arable land. To avoid repeating the mistakes of previous attempts to convert agricultural land to energy, arable land currently used for crops with known health hazards can be considered for conversion. Tobacco is the leading cause of avoidable death globally, and despite increasingly stringent controls on tobacco, economics provides an incentive to continue tobacco production. However, with the economics of PV ever improving, this study investigates the potential economic benefits of photovoltaic conversion of farms during tobacco's decline. This study analyzes key factors influencing conventional tobacco farming economics in the U.S. over a sensitivity of realistic future values. Then tobacco crop profit is compared to a sensitivity analysis covering the profits of solar PV farming on the same land. The results show that considering existing electric prices, escalation rates, and installed costs, PV farm substitution for tobacco farming makes economic sense in many U.S. cases already. In a case study of North Carolina, 30GW of PV power capacity was found to be economically viable on existing tobacco farms and if conversion took place over 2,000 premature deaths could be prevented from pollution reduction alone. This meets the State's peak summer loads. Land use policies are discussed to facilitate such land use conversions for the benefit of the economy, the environment and human health.
关键词: farming,solar photovoltaic,agriculture,tobacco,economic impact
更新于2025-09-11 14:15:04
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Engineered Nano-Structured Virus/ZnO Hybrid Materials with Dedicated Functional Properties
摘要: Bio-inspired mineralization for the production of new functional materials offers mild reaction conditions suitable to integrate biological templates and build hierarchically organized hybrid nanostructures with defined properties. In this respect, tobacco mosaic virus (TMV) stands out due to its unique structural dimensions. Here, we present a novel mineralization pathway for synthesis of virus-based ZnO hybrids with multifunctional properties. Wild-type TMV (wt-TMV), two TMV mutants (E50Q and TMV-Cys) and amino-functionalized self-assembled monolayers (NH2-SAMs), as a reference, were used as templates. This mineralization approach allows control of the particle size of the inorganic phase. Further, the virus contributes additionally to the texturing of ZnO. Field effect transistors (FETs) built from the hybrid films showed reproducible results at optimized conditions at close to ambient conditions and without post-treatment. The significantly reduced threshold voltage of the E50Q/ZnO FET compared to NH2-SAMs/ZnO FET points to the impact of the organic template on FET performance. Nacre-like virus-based ZnO multilayers and corresponding monolithic references were prepared. The mechanical properties by means of Young’s modulus, hardness and fracture toughness were determined and an increase of the mechanical performance by genetic modification was observed.
关键词: field effect transistors,Bio-inspired mineralization,ZnO hybrids,mechanical properties,tobacco mosaic virus
更新于2025-09-11 14:15:04
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Molecular Mechanics Simulations and Improved Tight-Binding Hamiltonians for Artificial Light Harvesting Systems: Predicting Geometric Distributions, Disorder, and Spectroscopy of Chromophores in a Protein Environment
摘要: We present molecular mechanics and spectroscopic calculations on prototype artificial light harvesting systems consisting of chromophores attached to a tobacco mosaic virus (TMV) protein scaffold. These systems have been synthesized and characterized spectroscopically, but information about the microscopic configurations and geometry of these TMV-templated chromophore assemblies is largely unknown. We use a Monte Carlo conformational search algorithm to determine the preferred positions and orientations of two chromophores, Coumarin 343 together with its linker, and Oregon Green 488, when these are attached at two different sites (104 and 123) on the TMV protein. The resulting geometric information shows that the extent of disorder and aggregation properties, and therefore the optical properties of the TMV-templated chromophore assembly, are highly dependent on the choice of chromophores and protein site to which they are bound. We used the results of the conformational search as geometric parameters together with an improved tight-binding Hamiltonian to simulate the linear absorption spectra and compare with experimental spectral measurements. The ideal dipole approximation to the Hamiltonian is not valid since the distance between chromophores can be very small. We found that using the geometries from the conformational search is necessary to reproduce the features of the experimental spectral peaks.
关键词: tobacco mosaic virus (TMV),linear absorption spectra,spectroscopic calculations,artificial light harvesting systems,Monte Carlo conformational search,tight-binding Hamiltonian,molecular mechanics,chromophores
更新于2025-09-10 09:29:36