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Keshan Disease (KD) is still a serious endemic public health problem in Northeast China. Previous studies have mainly focused on soil selenium content, staple food selenium content, and human selenium level, but there are few systematic studies on soil selenium existing forms and their migration from soil to crops and human body. In order to provide visualization of the spatial distribution characteristics and migration status of soil selenium content, and to provide a reference basis for the etiological analysis, prevention, control and elimination of KD, the 121183 topsoil soil (0-20 cm) geochemical data and 30295 soil parent material data which were collected and analysed by two-layer gridding method were used to infer the barrier factors in the migration and transformation of selenium from soil to crop and human body. The total selenium average of soil, staple food and human body be counted, the soil selenium existing forms and the correlation analysis between soil aluminum trioxide, iron trioxide were analysed.The average content of topsoil selenium in Northeast China was 0.20 mg/kg, which was significantly lower than the world soil selenium content average (0.4 mg/kg) and slightly lower than the Chinese soil selenium content average (0.239 mg/kg).The average selenium content of human hair is only 0.16mg/kg in Baiquan, Heilongjiang Province. Soil-forming parent material has some influence on soil selenium content, however, epigenetic enrichment is a more significant enrichment factor on soil selenium content. Soil selenium existing in humic acid state selenium, strongly organic bound state selenium, and the contents of selenite, selenate and methyl selenocysteine which are easily used by plants are all relatively high, correlation analysis between soil selenium content and organic matter, iron and aluminium oxides shows that soil organic matter and iron and aluminium oxides were the main barrier factors for selenium migration and transformation. Lower soil selenium content may be the basic factor for the biogeochemical deficiency of selenium, the sequestration of selenium by clay chemical constituents such as iron and aluminium oxides and soil organic matter is the direct factor for the biogeochemical low selenium in the Northeastern China, which increasing the risk of KD in the population. These research findings can provide more evidence to assess the elimination of biogeochemical aetiology of diseases from a selenium nutrient perspective.

Simulation wells with known parameters are vital in the study of anisotropic methods and logging equipment development. We propose a symmetrically partitioned isotropic (SPI) model, which can quantitatively control the parameters of equivalent circumferential anisotropy. First, the circumferential anisotropy of the flexural wave dispersion in the SPI model is analysed using a perturbation method, and the azimuth characteristics of the flexural wave and the wave velocity in the model are evaluated using a numerical simulation algorithm. Next, the circumferential anisotropy characteristics of the model are verified. Finally, the linear relationship between the fast and slow shear-wave velocities of the model and the model parameters is obtained through multiple linear regression analyses, and then, the equivalent circumferential anisotropy is predicted. The feasibility of the quantitative equivalent simulation of circumferential anisotropy media using the SPI model is verified.

Phosphatases catalyze the irreversible dephosphorylation of phosphate-containing compounds, and hence can be applied as the final enzymatic step for the synthesis of various biochemicals. However, the extensive substrate spectrums of phosphatases impose a great challenge for efficient biomanufacturing. Characterization of phosphatases is therefore of extreme importance. In this study, MmPase, a putative HAD phosphatase from Methanothermobacter marburgensis, was expressed, purified, and characterized. Recombinant MmPase was readily expressed in Escherichia coli, and required metal ions such as Mn

The transition between the planktonic state and the biofilm-associated state is a key developmental decision for pathogenic bacteria. Biofilm formation by Yersinia pestis is regulated by hmsHFRS genes (β-1, 6-N-acetyl-D-glucosamine synthesis operon) in its flea vector and in vitro. However, the mechanism of biofilm formation in Yersinia pseudotuberculosis remains elusive. In this study, we demonstrate that the LysR-type regulator RovM inversely regulates biofilm formation and motility in Y. pseudotuberculosis by acting as a transcriptional regulator of these two functions. RovM is strongly induced during growth in minimal media but strongly repressed in complex media. On one hand, RovM enhances bacterial motility by activating the expression of FlhDC, the master regulator of flagellar genes, via the recognition of an operator upstream of the flhDC promoter. On the other hand, RovM represses β-GlcNAc production under nutrition-limited conditions, negatively regulating hmsHFRS expression by directly binding to the −35 element of its promoter. Compared to wild-type bacteria, the rovM mutant established denser biofilms and caused more extensive mortality in mice and silkworm larvae. These results indicate that RovM acts as a molecular switch to coordinate the expression of genes involved in biofilm formation and motility in response to the availability of nutrients.

The type VI secretion system (T6SS) is a versatile molecular machinery widely distributed in Gram-negative bacteria. The activity of the T6SS is tightly regulated by various mechanisms, including quorum sensing (QS), iron concentration, and transcriptional regulators. Here we demonstrated that the stringent response regulator, RelA, contributes to bacterial resistance to multiple environmental stresses in Yersinia pseudotuberculosis. We also revealed that the stress resistance function of stringent response (SR) was partially mediated by the general stress response T6SS4 system. RelA positively regulates the expression of T6SS4 to combat various stresses in response to nutrition starvation collectively mediated by the RovM and RovA regulators. These findings revealed not only the important role of T6SS4 in SR induced stress resistance, but also a new pathway to regulate T6SS4 expression in response to starvation stress.

Micro-structural characteristics and electrical properties of an n-type GaN epilayer on Al2O3 irradiated by 290-MeV 238U32+ ions to various fluences were investigated using atomic force microscopy (AFM), scanning electron microscopy (SEM), high-resolution X-ray diffraction (HRXRD), and Raman scattering spectroscopy. AFM images show that the nano-hillocks generated, and the diameter and density of the nano-hillocks, increase obviously with increasing ion fluence, accompanied by an increase in surface roughness. SEM images display that the Al, O, and C elements appear on the GaN surface, along with a spiral-like, layered volcanic-cone structure formed at the highest-fluence irradiation. HRXRD reveals that the dislocation density increases, as the lattices gradually expand, and that Ga2O3 was produced with increasing ion fluence. Raman scattering spectra show that no N and Ga vacancies were produced, the free-carrier concentration decreases, while its mobility first increases and then exhibits a significant reduction with increasing ion fluence.

The application of the luxCDABE operon of the bioluminescent bacterium Photorhabdus luminescens as a reporter has been published for bacteria, yeast and mammalian cells. We report here the optimization of fused luxAB (the bacterial luciferase heterodimeric enzyme) expression, quantum yield and its application as a reporter gene in plant protoplasts. The fused luxAB gene was mutated by error prone PCR or chemical mutagenesis and screened for enhanced luciferase activity utilizing decanal as substrate. Positive luxAB mutants with superior quantum yield were subsequently shuffled by DNase I digestion and PCR assembly for generation of recombinants with additional increases in luciferase activity in bacteria. The coding sequence of the best recombinant, called eluxAB, was then optimized further to conform to Arabidopsis (Arabidopsis thaliana) codon usage. A plant expression vector of the final, optimized eluxAB gene (opt-eluxAB) was constructed and transformed into protoplasts of Arabidopsis and maize (Zea mays). Luciferase activity was dramatically increased for opt-eluxAB compared to the original luxAB in Arabidopsis and maize cells. The opt-eluxAB driven by two copies of the 35S promoter expresses significantly higher than that driven by a single copy. These results indicate that the eluxAB gene can be used as a reporter in plant protoplasts. To our knowledge, this is the first report to engineer the bacterium Photorhabdus luminescens luciferase luxAB as a reporter by directed evolution which paved the way for further improving the luxAB reporter in the future.

Protein-protein interactions are important for virtually every biological process, and a number of elegant approaches have been designed to detect and evaluate such interactions. However, few of these methods allow the detection of dynamic and real-time protein-protein interactions in bacteria. Here we describe a bioluminescence resonance energy transfer (BRET) system based on the bacterial luciferase LuxAB. We found that enhanced yellow fluorescent protein (eYFP) accepts the emission from LuxAB and emits yellow fluorescence. Importantly, BRET occurred when LuxAB and eYFP were fused, respectively, to the interacting protein pair FlgM and FliA. Furthermore, we observed sirolimus (i.e., rapamycin)-inducible interactions between FRB and FKBP12 and a dose-dependent abolishment of such interactions by FK506, the ligand of FKBP12. Using this system, we showed that osmotic stress or low pH efficiently induced multimerization of the regulatory protein OmpR and that the multimerization induced by low pH can be reversed by a neutralizing agent, further indicating the usefulness of this system in the measurement of dynamic interactions. This method can be adapted to analyze dynamic protein-protein interactions and the importance of such interactions in bacterial processes such as development and pathogenicity., IMPORTANCE Real-time measurement of protein-protein interactions in prokaryotes is highly desirable for determining the roles of protein complex in the development or virulence of bacteria, but methods that allow such measurement are not available. Here we describe the development of a bioluminescence resonance energy transfer (BRET) technology that meets this need. The use of endogenous excitation light in this strategy circumvents the requirement for the sophisticated instrument demanded by standard fluorescence resonance energy transfer (FRET). Furthermore, because the LuxAB substrate decanal is membrane permeable, the assay can be performed without lysing the bacterial cells, thus allowing the detection of protein-protein interactions in live bacterial cells. This BRET system added another useful tool to address important questions in microbiological studies.

Type VI secretion systems (T6SSs) are widespread multi-component machineries that translocate effectors into either eukaryotic or prokaryotic cells, for virulence or for interbacterial competition. Herein, we report that the T6SS-4 from Yersinia pseudotuberculosis displays an unexpected function in the transportation of Zn2+ to combat diverse stresses and host immunity. Environmental insults such as oxidative stress induce the expression of T6SS-4 via OxyR, the transcriptional factor that also regulates many oxidative response genes. Zinc transportation is achieved by T6SS-4-mediated translocation of a novel Zn2+-binding protein substrate YezP (YPK_3549), which has the capacity to rescue the sensitivity to oxidative stress exhibited by T6SS-4 mutants when added to extracellular milieu. Disruption of the classic zinc transporter ZnuABC together with T6SS-4 or yezP results in mutants that almost completely lost virulence against mice, further highlighting the importance of T6SS-4 in resistance to host immunity. These results assigned an unconventional role to T6SSs, which will lay the foundation for studying novel mechanisms of metal ion uptake by bacteria and the role of this process in their resistance to host immunity and survival in harmful environments., Author Summary One unique feature of type VI secretion system is the presence of multiple distinct systems in certain bacterial species. It is well established that some of these systems function to compete for their living niches among diverse bacterial species, whilst the activity of many such transporters remains unknown. Because metal ions are essential components to virtually all forms of life including bacteria, eukaryotic hosts have evolved complicated strategies to sequester metal ions, which constitute a major branch of their nutritional immunity. Therefore the ability to acquire metal ions is critical for bacterial virulence. This study reveals that the T6SS-4 of Yersinia pseudotuberculosis (Yptb) functions to import Zn2+ from the environment to mitigate the detrimental effects such as hydroxyl radicals induced by diverse stresses. Expression of the transporter is activated by multiple regulatory proteins, including OxyR and OmpR that sense diverse environmental cues. Zinc ion acquisition is achieved by translocating a Zn2+-binding substrate YezP, which is co-regulated with T6SS-4 by OxyR. Our results reveal a novel role for type VI secretion system, which is important in the study of the mechanism of metal ion acquisition by bacteria and the role of this process in bacterial pathogenesis and survival in detrimental environments.