Targeting‐induced local lesions in genomes (TILLING) is a powerful reverse‐genetics tool that enables high‐throughput screening of genomic variations in plants. Although TILLING has been developed ...for many diploid plants, the technology has been used in very few polyploid species due to their genomic complexity. Here, we established an efficient capillary electrophoresis‐based TILLING platform for allotetraploid cultivated tobacco (Nicotiana tabacum L.) using an ethyl methanesulfonate (EMS)‐mutagenized population of 1,536 individuals. We optimized the procedures for endonuclease preparation, leaf tissue sampling, DNA extraction, normalization, pooling, PCR amplification, heteroduplex formation, and capillary electrophoresis. In a test screen using seven target genes with eight PCR fragments, we obtained 118 mutants. The mutation density was estimated to be approximately one mutation per 106 kb on average. Phenotypic analyses showed that mutations in two heavy metal transporter genes, HMA2S and HMA4T, led to reduced accumulation of cadmium and zinc, which was confirmed independently using CRISPR/Cas9 to generate knockout mutants. Our results demonstrate that this powerful TILLING platform (available at http://www.croptilling.org) can be used in tobacco to facilitate functional genomics applications.
Tobacco has long been used to investigate development and hormonal responses. However, now it is used rarely due to the lack of proper genetic tools. Here we report the establishment of an efficient TILLING platform for tetraploid tobacco, which would be a powerful tool to generate mutants and new traits in tobacco.
: To investigate the effects of transcutaneous electrical stimulation (tES) on upper limb functional rehabilitation in a patient with traumatic brain injury (TBI), and to identify the optimum ...stimulation parameters of tES.
: A preliminary case study.
: Two successive interventions: Phase I-voluntary physical training (vPT) and Phase II - tES along with vPT (tES+vPT). tES was delivered at C3 and C6 cervical regions. Clinical assessments presented the variation of muscle tone and motor functions, before and after each training phase, and evaluated at 1-month follow up after the last intervention.
: Our results indicate that vPT alone contributed to a release of muscle spasticity of both arms of the patient with no significant improvement of hand function, while tES+vPT further reduced the spasticity of the left arm, and improved the voluntary motor function of both arms. The grip forces were also increased after the tES+vPT treatment. We found that 1 ms biphasic tES at 30 Hz produced optimum motor outputs.
: The study demonstrates, for the first time, the potential benefits of cervical tES in regard to improving upper limb motor functions in a patient with chronic TBI.
Mesenchymal stem cells (MSCs) hold great promise as therapeutic agents in regenerative medicine and autoimmune diseases, based on their differentiation abilities and immunosuppressive properties. ...However, the therapeutic applications raise a series of questions about the safety of culture-expanded MSCs for human use. This paper summarized recent findings about safety issues of MSCs, in particular their genetic stability in long-term in vitro expansion, their cryopreservation, banking, and the role of serum in the preparation of MSCs.
•We revealed the effects of global warming on the vegetation patterns based on a mathematical model.•We obtained the typical distributions of vegetation which is consistent with the results in Wuwei, ...China.•If the temperature increases 1.5 or more degrees, then the isolation of the vegetation pattern increases and thus desertification may emerge.•Vegetation biomass will increase as precipitation increases, but slow down with increased temperature.
Climate change has a great influence on the behavior of vegetation system in arid and semi-arid regions, yet the mechanisms are far from being well understood, especially on the pattern dynamics of vegetation. As a result, we pose a mathematical model in the form of reaction-diffusion equations to reveal the effects of global warming on the vegetation patterns. Based on mathematical analysis and numerical simulations, we obtain the typical distributions of vegetation which is consistent with the results in Wuwei, China. It was found that, if the temperature increases 1.5 or more degrees, then the isolation of the vegetation pattern increases and thus desertification may emerge. Meanwhile, we show that the vegetation biomass will increase as precipitation increases, but slow down with increased temperature. We also show the desertification degree as a function of precipitation. Our results may suggest that temperature and precipitation are key factors of climatic factors for vegetation formation.
Panax Notoginseng Saponins (PNS) is a formula of Chinese medicine commonly used for treating ischemia myocardial in China. However, its mechanism of action is yet unclear. This study investigated the ...effect and the mechanism of PNS on myocardial ischemia-reperfusion injury (MIRI) through the hypoxia-inducible factor 1α (HIF-1α)/bcl-2/adenovirus E1B19kDa-interacting protein3 (BNIP3) pathway of autophagy.
We constructed a rat model of myocardial injury and compared among 4 groups (n = 10, each): the sham-operated group (Sham), the ischemia-reperfusion group (IR), the PNS low-dose group, and the PNS high-dose group were pretreated with PNS (30 and 60 mg/kg, respectively). Serum creatine kinase, malonaldehyde (MDA), lactate dehydrogenase, myocardial tissue superoxide dismutase, and reactive oxygen species were detected in rats with myocardial ischemia-reperfusion after the intervention of PNS. The rat myocardial tissue was examined using hematoxylin and eosin (H&E) staining, and the mitochondria of myocardial cells were observed using transmission electron microscopy. The expressions of microtubule-associated protein light chain 3 (LC3), HIF-1α, BNIP3, Beclin-1, and autophagy-related gene-5 (Atg5) in rat myocardial tissue were detected using Western blotting.
The results showed that PNS was significantly protected against MIRI, as evidenced by the decreasing in the concentration of serum CK, MDA, lactate dehydrogenase, and myocardial tissue superoxide dismutase, reactive oxygen species, the attenuation of myocardial tissue histopathological changes and the mitochondrial damages of myocardial cells, and the increase of mitochondria autophagosome in myocardial cells. In addition, PNS significantly increased the expression of LC3 and the ratio of LC3II/LC3I in rat myocardial tissue. Moreover, PNS significantly increased the expression of HIF-1α, BNIP3, Atg5, and Beclin-1 in rat myocardial tissue.
The protective effect of PNS on MIRI was mainly due to its ability to enhance the mitochondrial autophagy of myocardial tissue through the HIF-1α/BNIP3 pathway.
14 new structures of isoxazole-moiety-containing quinazoline derivatives(3a~3n) were synthesized for the first time and characterized by IR, (1)H NMR, (13)C NMR, ESI-MS. Subsequently, their in vitro ...anticancer activity against A549, HCT116 and MCF-7 cell lines was preliminarily evaluated using the MTT method. Among them, most compounds showed good to excellent anticancer activity, especially 3d, 3i, 3k and 3m exhibited the more potent anticancer activity against A549, HCT116 and MCF-7 cell lines, which can be regarded as the promising drug candidates for development of anticancer drugs.
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•Se and SeO2 could deactivate NH3-SCR catalyst.•Se could be oxidized into Se4+ by V2O5.•SeO2 could be reduced to Se by NH3.•Se and SeO2 could sublimate from the catalyst to restore ...the activity.
Various trace elements in flue gas emitted from coal combustion can cause the deactivation of selective catalytic reduction (SCR) catalysts, yet rare studies have been reported on selenium species. Herein, the effects of Se and SeO2 on the denitrification performance of commercial V2O5-WO3/TiO2 SCR catalyst were investigated. Results indicated that the activities of Se and SeO2-doped catalysts were significantly inhibited at temperatures above 250 °C. During the NH3-SCR process, metallic Se was oxidized by V2O5 to generate Se4+ (SeO2 and binary oxides VxSeyOz), while part of Se4+ was reduced to Se by NH3. Se and SeO2 deactivated the catalyst by consuming V2O5 and NH3 during the NH3-SCR process, covering the active sites and relieved the redox performance of catalysts. With the sublimation of Se and SeO2, the deactivation effect on the catalyst was greatly relieved, while the presence of residual VxSeyOz resulted in the incomplete recovery of the catalyst activity.
Haloxylon ammodendron, a C4 perennial, succulent and xero-halophytic shrub, is highly resistant to harsh environments, therefore, exploring the stress resistance mechanism will be beneficial for the ...use of xerophytes to prevent desertification. To determine osmotic adjustment (OA) and antioxidase functions under simulated drought stress, 8-week-old seedlings were treated with sorbitol solutions to maintain osmotic potentials (Ψs) at a control and −0.5 and −1.0 MPa. Under −0.5 MPa osmotic stress, H. ammodendron stably maintained the water content of assimilating branches, a result that was not significantly different from the result of the control group. Moreover, the Ψs decreased significantly, which helped plants absorb water efficiently from the environment, as H. ammodendron accumulated massive osmotic regulators in its assimilating branches to adjust shoot Ψs. Specifically, the contribution of Na+ to shoot Ψs was up to 45%, and Na+ became the main osmotic regulator of OA. During the treatments, the content and contribution of K+ remained stable. However, the total contribution of three organic osmotic regulators (free proline, betaine and soluble sugar) was only 20%, and betaine was the main organic osmotic regulator, accounting for approximately 15% of the 20% contribution. Moreover, H. ammodendron seedlings presented strong antioxidases, especially when there was a high activity level of superoxide dismutase, and with an increase in treatment time and degree of osmotic stress, the activity of peroxidase and catalase increased significantly. Substantial accumulation of osmotic adjustment substances was an important strategy for H. ammodendron to cope with simulated drought stress, in particular, H. ammodendron absorbed much Na+ and transported Na+ into the assimilating branch for OA. The scavenging of reactive oxygen species by antioxidases was another adaptation strategy for H. ammodendron to adapt to simulated drought stress.
•Osmotic adjustment played important role in Haloxylon ammodendron under drought stress.•Na+ was accumulated largely in assimilating branch for osmotic adjustment.•Removing ROS through antioxidases was another important strategy.
Insight into wide aqueous pH influence on the electrocatalytic HER activity and stability of carbon nanotube-supported diiron dithiolato compound labeling as hybrid 2Fe2S-f-CNT are studied in this ...work.
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•New diiron dithiolato compounds 1–3 were prepared as a library of FeFe-hydrogenase models.•A new CNT-covalently-supported diiron compound labeling as 2Fe2S-f-CNT was constructed.•Electrocatalytic HER activity and stability of 2Fe2S-f-CNT is studied in wide pH aqueous media.•Electrochemical reconstruction of 2Fe2S-f-CNT leads to high HER activity in alkaline medium.•Electrochemical degradation of 2Fe2S-f-CNT results in low HER activity in neutral and acidic media.
Biomimetic chemistry on structure, function and external environment of FeFe-hydrogenases for hydrogen evolution reaction (HER) has shown a promising way to develop non-precious metal electrocatalysts for H2 production in the current Pt-dominated HER system. Herein, three new diiron dithiolato compounds {(μ-SCH2)2N(C6H4CH2CO2R)}Fe2(CO)6 (R = H (1), C6H4CHO-p (2), and C6H4Me-m (3)) were first prepared and identified as a library of FeFe-hydrogenase models in this work. Subsequently, the as-prepared diiron molecule 2 can be covalently attached onto carbon nanotubes (CNTs), resulting in the obtainment of target CNT-supported FeFe-hydrogenase model labeling as covalent hybrid 2Fe2S-f-CNT. Notably, the electrocatalytic HER performance and stability of the resulted hybrid 2Fe2S-f-CNT immobilized respectively on a gassy carbon (GC) electrode are systematically studied and compared in 0.1 M KOH (pH = 13), 0.1 M phosphate buffer (pH = 7), and 0.05 M H2SO4 (pH = 1) aqueous solutions by various spectroscopic and electrochemical techniques. The result has shown that the higher electrocatalytic HER activity is observed in alkaline solution (pH = 13) relative to neutral and acidic solutions (pH = 7, 1), being attributed to the different electrochemical HER processes of diiron cluster in wide pH aqueous media as revealed by post operando analysis.
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