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1.
Glob Chang Biol ; 2019 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-31418955

RESUMO

Sustainably feeding the growing population amid a changing climate and dwindling resources is a grand challenge facing mankind. Decades-long advancement in crop breeding has progressively elevated yield potential, markedly enhancing global food production capacity. However, relevant impact on reactive N (Nr) emissions associated with crop variety improvement has not been explicitly described. Here, we report multi-tiered evidence that newer and select maize, wheat, and rice varieties developed in China have the capacity to substantially lower Nr losses while producing more grain. First, we pooled studies featuring side-by-side comparison of different varieties, totaling 269 paired observations, to demonstrate that collectively, relatively newer varieties of maize, wheat, and rice had less Nr emissions (9.6-23.5%) while yielding more grains (7.3-11.2%) compared to older varieties under wide-ranging conditions. Next, we built an extended database (142 field studies with 833 observations) and comprehensively evaluated the Nr-loss reduction potential of newer varieties (2000 and after) vs. older ones (1985-1999). We found that newer varieties had Nr emission factors (EF, N loss as a percentage of N applied after correcting for background emissions) 18.2 to 75.7% less for N2 O, 18.3 to 75.7% less for NO3 - , and -8.5 to 22.8% less for NH3 , while producing more grains (16.0-24.4%). Individual varieties differed markedly in yield-emission scores. A nationwide farmer survey (2.47 million responses) indicated tremendous opportunities for a new way of management intervention. Coupling variety selection with sound N and other agronomic management can help lower N footprint while producing more grain. This article is protected by copyright. All rights reserved.

2.
J Am Chem Soc ; 141(20): 8128-8135, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31074995

RESUMO

Allosteric protein switches are key controllers of information and energy processing in living organisms and are desirable engineered control tools in synthetic systems. Here we present a generally applicable strategy for construction of allosteric signaling systems with inputs and outputs of choice. We demonstrate conversion of constitutively active enzymes into peptide-operated synthetic allosteric ON switches by insertion of a calmodulin domain into rationally selected sites. Switches based on EGFP, glucose dehydrogenase, NanoLuciferase, and dehydrofolate reductase required minimal optimization and demonstrated a dynamic response ranging from 1.8-fold in the former case to over 200-fold in the latter case. The peptidic nature of the calmodulin ligand enables incorporation of such synthetic switch modules into higher order sensory architectures. Here, a ligand-mediated increase in proximity of the allosteric switch and the engineered activator peptide modulates biosensor's activity. Created biosensors were used to measure concentrations of clinically relevant drugs and biomarkers in plasma, saliva, and urine with accuracy comparable to that of the currently used clinical diagnostic assays. The approach presented is generalizable as it allows rapid construction of efficient protein switches that convert binding of a broad range of analytes into a biochemical activity of choice enabling construction of artificial signaling and metabolic circuits of potentially unlimited complexity.

3.
Nat Microbiol ; 4(8): 1378-1388, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31110366

RESUMO

Mycobacterium tuberculosis (Mtb)-derived components are usually recognized by pattern recognition receptors to initiate a cascade of innate immune responses. One striking characteristic of Mtb is their utilization of different type VII secretion systems to secrete numerous proteins across their hydrophobic and highly impermeable cell walls, but whether and how these Mtb-secreted proteins are sensed by host immune system remains largely unknown. Here, we report that MPT53 (Rv2878c), a secreted disulfide-bond-forming-like protein of Mtb, directly interacts with TGF-ß-activated kinase 1 (TAK1) and activates TAK1 in a TLR2- or MyD88-independent manner. MPT53 induces disulfide bond formation at C210 on TAK1 to facilitate its interaction with TRAFs and TAB1, thus activating TAK1 to induce the expression of pro-inflammatory cytokines. Furthermore, MPT53 and its disulfide oxidoreductase activity is required for Mtb to induce the host inflammatory responses via TAK1. Our findings provide an alternative pathway for host signalling proteins to sense Mtb infection and may favour the improvement of current vaccination strategies.

4.
Sci Total Environ ; 657: 96-102, 2019 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-30537582

RESUMO

Global estimates of soil nitrate leaching of applied nitrogen (N) in agricultural production systems are not imprecise; however, the results of some field experiments have suggested that nitrate leaching responds exponentially rather than linearly to increasing N inputs. In this study, we compiled field data on nitrate leaching for 324 site-year combinations extracted from 86 peer-reviewed articles to test the hypothesis that in response to N fertilizer addition, soil nitrate leaching emission factors (EF) do not remain constant, but rather increase rapidly with increasing rates of N application. The averaged overall results showed that the proportional change in the EF response to increasing N input (ΔEF, %) was 0.042. Because this value was positive and significantly different from zero, our results demonstrate that EF is not constant, but rather increases with N addition. We compared our ΔEF response pattern with other constant EF patterns, and found that the 30% EF estimate of the Intergovernmental Panel on Climate Change and the 19% EF used in another published model were too high. A global estimate of soil nitrate leaching in arable uplands calculated with our ΔEF pattern was substantially lower than other estimates calculated with other procedures. In conclusion, our ΔEF pattern provided a globally applicable procedure for more accurate estimation of leaching loss and potential environmental costs incurred by various N application gradients.

5.
Mol Cancer Res ; 2018 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-30401720

RESUMO

The E-cadherin/ß-catenin signaling pathway plays a critical role in the maintenance of epithelial architecture and regulation of tumor progression. Normally, E-cadherin locates on the cell surface with its cytosolic domain linking to the actin cytoskeleton through interaction with catenins. Although the nuclear localization of E-cadherin has been frequently observed in various types of cancers, little is known regarding the functional consequences of its nuclear translocation. Here, we showed that in colorectal cancer samples and cell lines, E-cadherin localized in the nucleus; and the nuclear localization was mediated through protein interaction with CTNND1. In the nucleus, E-cadherin was acetylated by CBP at Lysine870 and Lysine871 in its ß-catenin binding domain, and the acetylation can be reversed by SIRT2. Acetylation of nuclear E-cadherin attenuated its interaction with ß-catenin, which therefore released ß-catenin from the complex, resulting in increased expression of its downstream genes and accelerated tumor growth and migration. Further study showed that acetylation level of nuclear E-cadherin had high prognostic significance in clinical colorectal samples. Taken together, our findings reveal a novel mechanism of tumor progression through post-translational modification of E-cadherin, which may serve as a potential drug target of tumor therapy. Implications: This finding that acetylation of nuclear E-cadherin regulates ß-catenin activity expands our understanding of the acetylation of E-cadherin promotes colorectal cancer cell growth and suggests novel therapeutic approaches of targeting acetylation in tumors.

6.
Sci Rep ; 8(1): 14762, 2018 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-30283043

RESUMO

To achieve food and water security, it is as important to close the water productivity (WP) gap (which was defined as the difference between the maximum attainable WP and the currently achieved WP at the field scale) as it is to close yield gaps. However, few studies have provided quantitative estimates of existing WP gaps and constraining factors for global maize production. Using a meta-analysis of 473 published studies covering 31 countries and 5,553 observations (932 site-years), we found the global average WP value for irrigated maize was 18.6 kg ha-1 mm-1. These WPs varied by factors such as seasonal precipitation, irrigation regimes, soil organic matter and soil pH. In current production systems, there exists a huge scope for improvement in maize WP, but the reported field experiments achieved only 20-46% of potential WP across all countries. Considering the future, raising WP to 85% of potential WP by 2050, a 100% increase in maize production could be achieved with 20% less planted area and 21% less water consumption than in 2005. Closing the WP gap may be critical to ensuring food security and achieving sustainable global agriculture.

7.
Nat Commun ; 9(1): 4295, 2018 10 16.
Artigo em Inglês | MEDLINE | ID: mdl-30327467

RESUMO

Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) kills millions every year, and there is urgent need to develop novel anti-TB agents due to the fast-growing of drug-resistant TB. Although autophagy regulates the intracellular survival of Mtb, the role of calcium (Ca2+) signaling in modulating autophagy during Mtb infection remains largely unknown. Here, we show that microRNA miR-27a is abundantly expressed in active TB patients, Mtb-infected mice and macrophages. The target of miR-27a is the ER-located Ca2+ transporter CACNA2D3. Targeting of this transporter leads to the downregulation of Ca2+ signaling, thus inhibiting autophagosome formation and promoting the intracellular survival of Mtb. Mice lacking of miR-27a and mice treated with an antagomir to miR-27a are more resistant to Mtb infection. Our findings reveal a strategy for Mtb to increase intracellular survival by manipulating the Ca2+-associated autophagy, and may also support the development of host-directed anti-TB therapeutic approaches.

8.
Virulence ; 9(1): 1468-1482, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30165798

RESUMO

The ability of Mycobacterium tuberculosis (M. tb) to survive and persist in the host for decades in an asymptomatic state is an important aspect of tuberculosis pathogenesis. Although adaptation to hypoxia is thought to play a prominent role underlying M. tb persistence, how the bacteria achieve this goal is largely unknown. Rv0081, a member of the DosR regulon, is induced at the early stage of hypoxia while Rv3334 is one of the enduring hypoxic response genes. In this study, we uncovered genetic interactions between these two transcription factors. RNA-seq analysis of ΔRv0081 and ΔRv3334 revealed that the gene expression profiles of these two mutants were highly similar. We also found that under hypoxia, Rv0081 positively regulated the expression of Rv3334 while Rv3334 repressed transcription of Rv0081. In addition, we demonstrated that Rv0081 formed dimer and bound to the promoter region of Rv3334. Taken together, these data suggest that Rv0081 and Rv3334 work in the same regulatory pathway and that Rv3334 functions immediately downstream of Rv0081. We also found that Rv3334 is a bona fide regulator of the enduring hypoxic response genes. Our study has uncovered a regulatory pathway that connects the early and the enduring hypoxic response, revealing a transcriptional cascade that coordinates the temporal response of M. tb to hypoxia.

9.
Nucleic Acids Res ; 46(12): 6387-6400, 2018 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-29846683

RESUMO

Sense codon reassignment to unnatural amino acids (uAAs) represents a powerful approach for introducing novel properties into polypeptides. The main obstacle to this approach is competition between the native isoacceptor tRNA(s) and orthogonal tRNA(s) for the reassigned codon. While several chromatographic and enzymatic procedures for selective deactivation of tRNA isoacceptors in cell-free translation systems exist, they are complex and not scalable. We designed a set of tRNA antisense oligonucleotides composed of either deoxy-, ribo- or 2'-O-methyl ribonucleotides and tested their ability to efficiently complex tRNAs of choice. Methylated oligonucleotides targeting sequence between the anticodon and variable loop of tRNASerGCU displayed subnanomolar binding affinity with slow dissociation kinetics. Such oligonucleotides efficiently and selectively sequestered native tRNASerGCU directly in translation-competent Escherichia coli S30 lysate, thereby, abrogating its translational activity and liberating the AGU/AGC codons. Expression of eGFP protein from the template harboring a single reassignable AGU codon in tRNASerGCU-depleted E. coli lysate allowed its homogeneous modification with n-propargyl-l-lysine or p-azido-l-phenylalanine. The strategy developed here is generic, as demonstrated by sequestration of tRNAArgCCU isoacceptor in E. coli translation system. Furthermore, this method is likely to be species-independent and was successfully applied to the eukaryotic Leishmania tarentolae in vitro translation system. This approach represents a new direction in genetic code reassignment with numerous practical applications.

10.
Emerg Microbes Infect ; 7(1): 34, 2018 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-29559631

RESUMO

Tuberculosis caused by Mycobacterium tuberculosis (Mtb) infection remains a large global public health problem. One striking characteristic of Mtb is its ability to adapt to hypoxia and trigger the ensuing transition to a dormant state for persistent infection, but how the hypoxia response of Mtb is regulated remains largely unknown. Here we performed a quantitative acetylome analysis to compare the acetylation profile of Mtb under aeration and hypoxia, and showed that 377 acetylation sites in 269 Mtb proteins were significantly changed under hypoxia. In particular, deacetylation of dormancy survival regulator (DosR) at K182 promoted the hypoxia response in Mtb and enhanced the transcription of DosR-targeted genes. Mechanistically, recombinant DosRK182R protein demonstrated enhanced DNA-binding activity in comparison with DosRK182Q protein. Moreover, Rv0998 was identified as an acetyltransferase that mediates the acetylation of DosR at K182. Deletion of Rv0998 also promoted the adaptation of Mtb to hypoxia and the transcription of DosR-targeted genes. Mice infected with an Mtb strain containing acetylation-defective DosRK182R had much lower bacterial counts and less severe histopathological impairments compared with those infected with the wild-type strain. Our findings suggest that hypoxia induces the deacetylation of DosR, which in turn increases its DNA-binding ability to promote the transcription of target genes, allowing Mtb to shift to dormancy under hypoxia.


Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Lisina/metabolismo , Mycobacterium tuberculosis/metabolismo , Oxigênio/metabolismo , Proteínas Quinases/química , Proteínas Quinases/metabolismo , Tuberculose/microbiologia , Acetilação , Motivos de Aminoácidos , Proteínas de Bactérias/genética , Regulação Bacteriana da Expressão Gênica , Humanos , Mycobacterium tuberculosis/química , Mycobacterium tuberculosis/genética , Proteínas Quinases/genética
11.
Nature ; 555(7696): 363-366, 2018 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-29513654

RESUMO

Sustainably feeding a growing population is a grand challenge, and one that is particularly difficult in regions that are dominated by smallholder farming. Despite local successes, mobilizing vast smallholder communities with science- and evidence-based management practices to simultaneously address production and pollution problems has been infeasible. Here we report the outcome of concerted efforts in engaging millions of Chinese smallholder farmers to adopt enhanced management practices for greater yield and environmental performance. First, we conducted field trials across China's major agroecological zones to develop locally applicable recommendations using a comprehensive decision-support program. Engaging farmers to adopt those recommendations involved the collaboration of a core network of 1,152 researchers with numerous extension agents and agribusiness personnel. From 2005 to 2015, about 20.9 million farmers in 452 counties adopted enhanced management practices in fields with a total of 37.7 million cumulative hectares over the years. Average yields (maize, rice and wheat) increased by 10.8-11.5%, generating a net grain output of 33 million tonnes (Mt). At the same time, application of nitrogen decreased by 14.7-18.1%, saving 1.2 Mt of nitrogen fertilizers. The increased grain output and decreased nitrogen fertilizer use were equivalent to US$12.2 billion. Estimated reactive nitrogen losses averaged 4.5-4.7 kg nitrogen per Megagram (Mg) with the intervention compared to 6.0-6.4 kg nitrogen per Mg without. Greenhouse gas emissions were 328 kg, 812 kg and 434 kg CO2 equivalent per Mg of maize, rice and wheat produced, respectively, compared to 422 kg, 941 kg and 549 kg CO2 equivalent per Mg without the intervention. On the basis of a large-scale survey (8.6 million farmer participants) and scenario analyses, we further demonstrate the potential impacts of implementing the enhanced management practices on China's food security and sustainability outlook.


Assuntos
Agricultura/métodos , Conservação dos Recursos Naturais , Produtos Agrícolas/crescimento & desenvolvimento , Eficiência Organizacional , Fazendeiros , China , Técnicas de Apoio para a Decisão , Grão Comestível/crescimento & desenvolvimento , Política Ambiental , Fertilizantes/estatística & dados numéricos , Abastecimento de Alimentos/métodos , Efeito Estufa , Nitrogênio/metabolismo , Oryza/crescimento & desenvolvimento , Triticum/crescimento & desenvolvimento , Zea mays/crescimento & desenvolvimento
12.
Sci Rep ; 7(1): 7016, 2017 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-28765540

RESUMO

Increasing grain zinc (Zn) concentration of cereals for minimizing Zn malnutrition in two billion people represents an important global humanitarian challenge. Grain Zn in field-grown wheat at the global scale ranges from 20.4 to 30.5 mg kg-1, showing a solid gap to the biofortification target for human health (40 mg kg-1). Through a group of field experiments, we found that the low grain Zn was not closely linked to historical replacements of varieties during the Green Revolution, but greatly aggravated by phosphorus (P) overuse or insufficient nitrogen (N) application. We also conducted a total of 320-pair plots field experiments and found an average increase of 10.5 mg kg-1 by foliar Zn application. We conclude that an integrated strategy, including not only Zn-responsive genotypes, but of a similar importance, Zn application and field N and P management, are required to harvest more grain Zn and meanwhile ensure better yield in wheat-dominant areas.

13.
J Immunol ; 198(12): 4772-4780, 2017 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-28507027

RESUMO

Tuberculosis caused by Mycobacterium tuberculosis continues to pose a serious global health threat. The attenuated Mycobacterium bovis bacillus Calmette-Guérin, as the only licensed vaccine, has limited protective efficacy against TB. The development of more effective antituberculosis vaccines is urgent and demands for further identification and understanding of M. tuberculosis Ags. MPT83 (Rv2873), a secreted mycobacterial lipoprotein, has been applied into subunit vaccine development and shown protective effects against M. tuberculosis infection in animals; however, the understanding of the underlying mechanism is limited. In present study, we systematically studied the effect of MPT83 on macrophage apoptosis by constructing Mycobacterium smegmatis strain overexpressing MPT83 (MS_MPT83) and purifying rMPT83 protein. We found that MPT83 induced apoptosis in both human and mouse macrophages. MPT83 induced cyclooxygenase-2 (COX-2) expression at both the transcriptional and protein levels in macrophages, whereas silencing or inhibiting COX-2 blocked rMPT83-induced apoptosis or the enhanced apoptotic response to MS_MPT83 in comparison with M. smegmatis transfected with pMV261 vector (MS_Vec), indicating that COX-2 is required for MPT83-induced apoptosis. Additionally, tlr2 deficiency led to significant reduction of COX-2 expression, accompanied by less apoptosis in macrophages stimulated with rMPT83 or infected with MS_MPT83. Moreover, the activation of p38 accounted for MPT83-induced COX-2 expression. Finally, lower bacteria burdens in the lungs and spleens and enhanced survival were observed in mice i.v. infected with MS_MPT83 compared with MS_Vec. Taken together, our results established a proapoptotic effect of MPT83 and identified the TLR2/p38/COX-2 axis in MPT83-induced macrophage apoptosis.


Assuntos
Antígenos de Bactérias/fisiologia , Apoptose , Proteínas de Bactérias/fisiologia , Ciclo-Oxigenase 2/metabolismo , Macrófagos/microbiologia , Macrófagos/fisiologia , Proteínas de Membrana/fisiologia , Transdução de Sinais/imunologia , Receptor 2 Toll-Like/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Antígenos de Bactérias/genética , Antígenos de Bactérias/isolamento & purificação , Apoptose/efeitos dos fármacos , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Ciclo-Oxigenase 2/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Proteínas de Membrana/genética , Proteínas de Membrana/isolamento & purificação , Camundongos , Mycobacterium smegmatis/imunologia , Fosforilação , Receptor 2 Toll-Like/imunologia
14.
Sci Rep ; 7: 41587, 2017 02 03.
Artigo em Inglês | MEDLINE | ID: mdl-28155860

RESUMO

The food supply is being increasingly challenged by climate change and water scarcity. However, incremental changes in traditional cropping systems have achieved only limited success in meeting these multiple challenges. In this study, we applied a systematic approach, using model simulation and data from two groups of field studies conducted in the North China Plain, to develop a new cropping system that improves yield and uses water in a sustainable manner. Due to significant warming, we identified a double-maize (M-M; Zea mays L.) cropping system that replaced the traditional winter wheat (Triticum aestivum L.) -summer maize system. The M-M system improved yield by 14-31% compared with the conventionally managed wheat-maize system, and achieved similar yield compared with the incrementally adapted wheat-maize system with the optimized cultivars, planting dates, planting density and water management. More importantly, water usage was lower in the M-M system than in the wheat-maize system, and the rate of water usage was sustainable (net groundwater usage was ≤150 mm yr-1). Our study indicated that systematic assessment of adaptation and cropping system scale have great potential to address the multiple food supply challenges under changing climatic conditions.

15.
PLoS One ; 12(2): e0172717, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28234970

RESUMO

Understanding the physiological changes associated with high grain yield and high N use efficiency (NUE) is important when increasing the plant density and N rate to develop optimal agronomic management. We tested the hypothesis that high plant densities resulting in crowding stress reduce the ability of plants to use the N supply post-silking, thus decreasing the grain yield and NUE. In 2013 and 2014, a field experiment, with five N-application rates and three plant densities (6.0, 7.5, and 9.0 plants m-2), was conducted in the North China Plain (NCP). The calculated maximum grain yield and agronomic use efficiency (AEN) at a density of 7.5 plants m-2 were 12.4 Mg ha-1 and 39.3 kg kg-1, respectively, which were significantly higher than the values obtained at densities of 6.0 (11.3 Mg ha-1 and 30.2 kg kg-1) and 9.0 plant m-2 (11.7 Mg ha-1 and 27.8 kg kg-1). A high plant density of 9.0 plants m-2 decreased the post-silking N accumulation, leaf N concentration and net photosynthesis, which reduced the post-silking dry matter production, resulting in a low yield and NUE. Although a relatively low grain yield was observed at a density of 9.0 plants m-2, the optimal N rate increased from 150 to 186 kg N ha-1 at a density of 7.5 plants m-2. These results indicate that high plant densities with crowding stress reduce the ability of plants to use soil N during the post-silking period, and high rate of N fertilizer was needed to increase grain yield. We conclude that selecting the appropriate plant density combined with optimal N management could increase grain yields and the NUE in the NCP.


Assuntos
Fixação de Nitrogênio/fisiologia , Fotossíntese/fisiologia , Fenômenos Fisiológicos Vegetais , Zea mays/crescimento & desenvolvimento , Agricultura , Fertilizantes , Nitrogênio/metabolismo , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Densidade Demográfica , Solo/química , Luz Solar , Zea mays/metabolismo
16.
ACS Synth Biol ; 6(3): 535-544, 2017 03 17.
Artigo em Inglês | MEDLINE | ID: mdl-27966891

RESUMO

Incorporation of unnatural amino acids (uAAs) via codon reassignment is a powerful approach for introducing novel chemical and biological properties to synthesized polypeptides. However, the site-selective incorporation of multiple uAAs into polypeptides is hampered by the limited number of reassignable nonsense codons. This challenge is addressed in the current work by developing Escherichia coli in vitro translation system depleted of specific endogenous tRNAs. The translational activity in this system is dependent on the addition of synthetic tRNAs for the chosen sense codon. This allows site-selective uAA incorporation via addition of tRNAs pre- or cotranslationally charged with uAA. We demonstrate the utility of this system by incorporating the BODIPY fluorophore into the unique AGG codon of the calmodulin(CaM) open reading frame using in vitro precharged BODIPY-tRNACysCCU. The deacylated tRNACysCCU is a poor substrate for Cysteinyl-tRNA synthetase, which ensures low background incorporation of Cys into the chosen codon. Simultaneously, p-azidophenylalanine mediated amber-codon suppression and its post-translational conjugation to tetramethylrhodamine dibenzocyclooctyne (TAMRA-DIBO) were performed on the same polypeptide. This simple and robust approach takes advantage of the compatibility of BODIPY fluorophore with the translational machinery and thus requires only one post-translational derivatization step to introduce two fluorescent labels. Using this approach, we obtained CaM nearly homogeneously labeled with two FRET-forming fluorophores. Single molecule FRET analysis revealed dramatic changes in the conformation of the CaM probe upon its exposure to Ca2+ or a chelating agent. The presented approach is applicable to other sense codons and can be directly transferred to eukaryotic cell-free systems.


Assuntos
Aminoácidos/genética , Códon sem Sentido/genética , Biossíntese de Proteínas/genética , Processamento de Proteína Pós-Traducional/genética , Aminoacil-tRNA Sintetases/metabolismo , Azidas/farmacologia , Cálcio/metabolismo , Sistema Livre de Células/efeitos dos fármacos , Sistema Livre de Células/metabolismo , Códon de Terminação/genética , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Escherichia coli/metabolismo , Eucariotos/efeitos dos fármacos , Eucariotos/genética , Eucariotos/metabolismo , Fenilalanina/análogos & derivados , Fenilalanina/farmacologia , Biossíntese de Proteínas/efeitos dos fármacos , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , RNA de Transferência/genética
17.
Nature ; 537(7622): 671-674, 2016 09 29.
Artigo em Inglês | MEDLINE | ID: mdl-27602513

RESUMO

Sustainably feeding the world's growing population is a challenge, and closing yield gaps (that is, differences between farmers' yields and what are attainable for a given region) is a vital strategy to address this challenge. The magnitude of yield gaps is particularly large in developing countries where smallholder farming dominates the agricultural landscape. Many factors and constraints interact to limit yields, and progress in problem-solving to bring about changes at the ground level is rare. Here we present an innovative approach for enabling smallholders to achieve yield and economic gains sustainably via the Science and Technology Backyard (STB) platform. STB involves agricultural scientists living in villages among farmers, advancing participatory innovation and technology transfer, and garnering public and private support. We identified multifaceted yield-limiting factors involving agronomic, infrastructural, and socioeconomic conditions. When these limitations and farmers' concerns were addressed, the farmers adopted recommended management practices, thereby improving production outcomes. In one region in China, the five-year average yield increased from 67.9% of the attainable level to 97.0% among 71 leading farmers, and from 62.8% to 79.6% countywide (93,074 households); this was accompanied by resource and economic benefits.


Assuntos
Produção Agrícola/métodos , Produção Agrícola/estatística & dados numéricos , Fazendeiros/estatística & dados numéricos , Triticum/crescimento & desenvolvimento , Zea mays/crescimento & desenvolvimento , China , Mudança Climática , Produção Agrícola/economia , Ecologia , Política Ambiental , Nitrogênio , População Rural , Sementes , Fatores Socioeconômicos , Fatores de Tempo , Água
18.
Front Microbiol ; 7: 618, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27199947

RESUMO

BACKGROUND: Failure to early detect multidrug-resistant tuberculosis (MDR-TB) results in treatment failure and poor clinical outcomes, and highlights the need to rapidly detect resistance to rifampicin (RIF) and isoniazid (INH). METHODS: In Multi-Fluorescence quantitative Real-Time PCR (MF-qRT-PCR) assay, 10 probes labeled with four kinds of fluorophores were designed to detect the mutations in regions of rpoB, katG, mabA-inhA, oxyR-ahpC, and rrs. The efficiency of MF-qRT-PCR assay was tested using 261 bacterial isolates and 33 clinical sputum specimens. Among these samples, 227 Mycobacterium tuberculosis isolates were analyzed using drug susceptibility testing (DST), DNA sequencing and MF-qRT-PCR assay. RESULTS: Compared with DST, MF-qRT-PCR sensitivity and specificity for RIF-resistance were 94.6 and 100%, respectively. And the detection sensitivity and specificity for INH-resistance were 85.9 and 95.3%, respectively. Compared with DNA sequencing, the sensitivity and specificity of our assay were 97.2 and 100% for RIF-resistance and 97.9 and 96.4% for INH-resistance. Compared with Phenotypic strain identification, MF-qRT-PCR can distinguish 227 M. tuberculosis complexes (MTC) from 34 Non-tuberculous mycobacteria (NTM) isolates with 100% accuracy rate. CONCLUSIONS: MF-qRT-PCR assay was an efficient, accurate, reliable, and easy-operated method for detection of RIF and INH-resistance, and distinction of MTC and NTM of clinical isolates.

19.
PLoS One ; 11(2): e0148965, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26886880

RESUMO

The transcription regulatory system of Mycobacterium tuberculosis (M. tb) remains incompletely understood. In this study, we have applied the eGLECLUBS algorithm to a group of related prokaryotic genomes for de novo genome-wide prediction of cis-regulatory binding sites (CRBSs) in M. tb H37Rv. The top 250 clusters from our prediction recovered 83.3% (50/60) of all known CRBSs in extracted inter-operonic sequences of this strain. We further demonstrated that the integration of our prediction results with the ChIP-Seq datasets is very effective in identifying true binding sites of TFs. Using electrophoretic mobility shift assays and real-time RT-PCR, we experimentally verified our prediction of CRBSs for Rv0081, an important transcription factor thought to be involved in regulation of M. tb under hypoxia.


Assuntos
Genoma Bacteriano , Mycobacterium tuberculosis/genética , Sequências Reguladoras de Ácido Nucleico/genética , Sítios de Ligação/genética , Imunoprecipitação da Cromatina , Bases de Dados Genéticas , Ensaio de Desvio de Mobilidade Eletroforética , Regulação Bacteriana da Expressão Gênica , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Software , Fatores de Transcrição/metabolismo , Regulação para Cima/genética
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