Dynamic Response of the cbbL Carbon Sequestration Microbial Community to Wetland Type in Qinghai Lake.

The soil carbon storage in the Qinghai-Tibet Plateau wetlands is affected by microbiota and wetland types, but the response mechanisms of carbon sequestration microorganisms on the Qinghai-Tibet Plateau to different wetland types are still poorly described. To explore the differences in carbon sequestration microbial communities in different wetlands and the main influencing factors, this study took a marsh wetland, river source wetland and lakeside wetland of Qinghai Lake as the research objects and used high-throughput sequencing to study the functional gene, cbbL, of carbon sequestration microorganisms. The results showed that the dominant bacterial group of carbon sequestration microorganisms in marsh and river source wetlands was Proteobacteria, and the dominant bacterial group in the lakeside wetland was Cyanobacteria. The alpha diversity, relative abundance of Proteobacteria and total carbon content were the highest in the marsh wetland, followed by the river source wetland, and they were the lowest in the lakeside wetland. In addition, the physical and chemical characteristics of the three wetland types were significantly different, and the soil temperature and moisture and total carbon content were the most important factors affecting the community structures of carbon-sequestering microorganisms. There was little difference in the total nitrogen contents between the marsh wetland and river source wetland. However, the total nitrogen content was also an important factor affecting the diversity of the carbon sequestration microbial community. In summary, the wetland type significantly affects the process of soil carbon sequestration. Compared with the riverhead and lakeside wetlands, the marsh wetland has the highest carbon storage.

Hormetic effects of EGC and EGCG on CES1 activity and its rescue from oxidative stress in rat liver S9.

Carboxylesterase 1 (CES1) is a hydrolytic enzyme that plays an important role in the activation or deactivation of many therapeutic agents, thus affecting their pharmacokinetic and pharmacodynamic outcomes. Using rat liver S9 as an enzyme source and enalapril as a CES1 substrate, the present study examined effects of a number of flavonoids on the formation of enalaprilat (the active form of enalapril) produced by CES1-mediated hydrolysis. While a majority of flavonoids tested showed inhibition on CES1, an unexpected hormetic effect was observed for epigallocatechin (EGC) and epigallocatechin gallate (EGCG), i.e., stimulatory effect at low concentrations and enzyme inhibition at high concentrations. Further experiments revealed that oxidative stress caused by hydrogen peroxide, arachidonic acid plus iron, and oxidized low density lipoproteins (oxLOL) reduced CES1 activity in rat liver S9 and the loss of CES1 enzyme activity could be rescued largely by EGC or EGCG. In contrast, such effects were minimal in human liver S9, probably due to the presence of a higher ratio of reduced vs oxidized forms of glutathione. The above findings suggest that the polyphenolic nature of EGC or EGCG might be responsible for rescuing CES1 activity under oxidative stress. Because of the importance of CES1 in drug activation or deactivation and rat liver S9 as a versatile in vitro system used for drug metabolism studies and drug safety assessment, caution should be exercised to avoid potential biases for data interpretation and decision making when CES1 activity in rat liver S9 is evaluated with dependency on experimental conditions.

Antitumor activity of mianserin (a tetracyclic antidepressant) primarily driven by the inhibition of SLC1A5-mediated glutamine transport.

Targeting tumor metabolic vulnerabilities such as "glutamine addiction" has become an attractive approach for the discovery of novel antitumor agents. Among various mechanisms explored, SLC1A5, a membrane transporter that plays an important role in glutamine cellular uptake, represents a viable target to interfere with tumor's ability to acquire critical nutrients during proliferation. In the present study, a stably transfected HEK293 cell line with human SLC1A5 (HEK293-SLC1A5) was established for the screening and identification of small molecule SLC1A5 inhibitors. This in vitro system, in conjunction with direct measurement of SLC1A5-mediated L-glutamine-2,3,3,4,4-D5 (substrate) uptake, was practical and efficient in ensuring the specificity of SLC1A5 inhibition. Among a group of diverse compounds tested, mianserin (a tetracyclic antidepressant) demonstrated a marked inhibition of SLC1A5-mediated glutamine uptake. Subsequent investigations using SW480 cells demonstrated that mianserin was capable of inhibiting SW480 tumor growth both in vitro and in vivo, and the in vivo antitumor efficacy was correlated to the reduction of glutamine concentrations in tumor tissues. Computational analysis revealed that hydrophobic interactions between SLC1A5 and its inhibitors could be a critical factor in drug design. Taken together, the current findings confirmed the feasibility of targeting SLC1A5-mediated glutamine uptake as a novel approach for antitumor intervention. It is anticipated that structural insights obtained based on homology modeling would lead to the discovery of more potent and specific SLC1A5 inhibitors for clinical development.

Recent advances of using personal glucose meter as a biosensor readout for non-glucose targets.

Background: Personal glucose meter (PGM) has become the most successful biosensor in past decades due to its advantages of small size, convenient operation, and low cost. To take advantage of many years of research and development of PGMs, new signal transduction methods has been developed to expand the PGM from simple monitoring blood glucose to detection of numerous non-glucose targets. Objectives: This review summarizes recent advance of PGM-based biosensors for non-glucose targets including signal transduction, signal amplification and target molecule recognition and analysis. Current challenges and future directions are also discussed. Conclusion: PGM can be used as biosensor readout to detect various non-glucose targets from metal ion, small molecule to protein and even living organisms such as bacteria and other pathogens by using different signal transduction elements such as invertase and amylase, and different signal amplification methods such as nanomaterials, nucleic acid reaction, liposome encapsulation, hydrogel trapping, DNAzyme amplification and biotin-streptavidin reaction.

[Response of Maize to Lead Stress and Relevant Chemical Forms of Lead].

A sand-culture experiment was carried out to research the differences in lead (Pb) chemical-form among different maize varieties for roots and shoots under Pb stress, and further investigate into the mechanism of maize endurance to resist Pb. The results showed that the wheat varieties of Zhengdan 958 and Longping 206 have the maximum Pb tolerance, whereas Lianchuang 5's tolerance of Pb was the minimum. Pb form in roots and shoots were mainly harmfulness HAc-extractable and HCl-extractable, accounting for a high proportion of 60% - 87%. Moreover, these values in roots were slightly higher than those in shoots. And concentrations of alcohol-extractable and water-extractable Pb accounted for the proportion of 6% - 20%. Under 100 mg x L(-1) Pb stress, the alcohol-extractable together with water-extractable Pb content in shoots of Longping 206 was the lowest (0.52 mg x kg(-1)), and that of in Zhengdan 958 shoots was 0.93 mg x kg(-1) which was also very low. However, for Lianchuang 5, the content could reach 2.78 mg x kg(-1). Under the stress of 800 mg x L(-1) Pb, content of alcohol-extractable together with water-extractable Pb in Zhengdan 958 shoots was 2.41 mg x kg(-1), which was still the lowest. These were probable reasons why Zhengdan 958 was more resistant to Pb stress than other varieties. Tolerance of Zhengdan 958 to Pb stress was related to it's strong ability to convert toxic Pb into non-toxic Pb.

Genetic engineering of the Xa10 promoter for broad-spectrum and durable resistance to Xanthomonas oryzae pv. oryzae.

Many pathovars of plant pathogenic bacteria Xanthomonas species inject transcription activator-like (TAL) effectors into plant host cells to promote disease susceptibility or trigger disease resistance. The rice TAL effector-dependent disease resistance gene Xa10 confers narrow-spectrum race-specific resistance to Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of bacterial blight disease in rice. To generate broad-spectrum and durable resistance to Xoo, we developed a modified Xa10 gene, designated as Xa10(E5) . Xa10(E5) has an EBE-amended promoter containing 5 tandemly arranged EBEs each responding specifically to a corresponding virulent or avirulent TAL effector and a stable transgenic rice line containing Xa10(E5) was generated in the cultivar Nipponbare. The Xa10(E5) gene was specifically induced by Xoo strains that harbour the corresponding TAL effectors and conferred TAL effector-dependent resistance to the pathogens at all developmental stages of rice. Further disease evaluation demonstrated that the Xa10(E5) gene in either Nipponbare or 9311 genetic backgrounds provided broad-spectrum disease resistance to 27 of the 28 Xoo strains collected from 11 countries. The development of Xa10(E5) and transgenic rice lines provides new genetic materials for molecular breeding of rice for broad-spectrum and durable disease resistance to bacterial blight.

The rice TAL effector-dependent resistance protein XA10 triggers cell death and calcium depletion in the endoplasmic reticulum.

The recognition between disease resistance (R) genes in plants and their cognate avirulence (Avr) genes in pathogens can produce a hypersensitive response of localized programmed cell death. However, our knowledge of the early signaling events of the R gene-mediated hypersensitive response in plants remains limited. Here, we report the cloning and characterization of Xa10, a transcription activator-like (TAL) effector-dependent R gene for resistance to bacterial blight in rice (Oryza sativa). Xa10 contains a binding element for the TAL effector AvrXa10 (EBEAvrXa10) in its promoter, and AvrXa10 specifically induces Xa10 expression. Expression of Xa10 induces programmed cell death in rice, Nicotiana benthamiana, and mammalian HeLa cells. The Xa10 gene product XA10 localizes as hexamers in the endoplasmic reticulum (ER) and is associated with ER Ca(2+) depletion in plant and HeLa cells. XA10 variants that abolish programmed cell death and ER Ca(2+) depletion in N. benthamiana and HeLa cells also abolish disease resistance in rice. We propose that XA10 is an inducible, intrinsic terminator protein that triggers programmed cell death by a conserved mechanism involving disruption of the ER and cellular Ca(2+) homeostasis.

Production of marker-free transgenic rice expressing tissue-specific Bt gene.

The hybrid Bacillus thuringiensis (Bt) δ-endotoxin gene Cry1Ab/Ac was used to develop a transgenic Bt rice (Oryza sativa L.) targeting lepidopteran insects of rice. Here, we show the production of a marker-free and tissue-specific expressing transgenic Bt rice line L24 using Agrobacterium-mediated transformation and a chemically regulated, Cre/loxP-mediated DNA recombination system. L24 carries a single copy of marker-free T-DNA that contains the Cry1Ab/Ac gene driven by a maize phosphoenolpyruvate carboxylase (PEPC) gene promoter. The marker-free T-DNA was integrated into the 3' untranslated region of rice gene Os01g0154500 on the short arm of chromosome 1. Compared to the constitutive and non-specific expression of the P (Actin1):Cry1Ab/Ac:T (Nos) gene in the control Bt rice line T51-1, the P ( Pepc ):Cry1Ab/Ac:T (Nos ) gene was detected only in the leaf and stem tissues of L24. More importantly, compared to high levels of CRY1Ab/Ac proteins accumulated in T51-1 seeds, the CRY1Ab/Ac proteins were not detectable in L24 seeds by Western blot analysis. As demonstrated by insect bioassay, L24 provided similar level of resistance to rice leaffolder (Cnaphalocrocis medinalis) as T51-1. The marker-free transgenic line L24 can be used directly in rice breeding for insect resistance to lepidopteran insects where absence of Bt toxin protein in the seed is highly desirable.