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CRISPR-Cas systems adapt “memories” via spacers from viruses and plasmids to develop adaptive immunity against mobile genetic elements. Mature CRISPR RNAs guide CRISPR-associated nucleases to site-specifically cleave target DNA or RNA, providing an efficient genome engineering tool for organisms of all three kingdoms. Cas9, Cas12, and Cas13 are single proteins with multiple domains that are the most widely used CRISPR nucleases of the Class 2 system. However, these CRISPR endonucleases are large in size, leading to difficulty for manipulation and toxicity for cells. Most archaeal genomes and half of the bacterial genomes encode different types of CRISPR-Cas systems. Therefore, developing endogenous CRISPR-Cas systems-based genome editing will simplify manipulations and increase editing efficiency in prokaryotic cells. Here, we review the current applications and discuss the prospects of using endogenous CRISPR nucleases for genome engineering and CRISPR-based antimicrobials.

In Klebsiella pneumoniae CG43S3, deletion of the response regulator gene rcsB reduced the capsular polysaccharide amount and survival on exposure to acid stress. A comparison of the pH 4.4-induced proteomes between CG43S3 and CG43S3ΔrcsB revealed numerous differentially expressed proteins and one of them, YfdX, which has recently been reported as a periplasmic protein, was absent in CG43S3ΔrcsB. Acid survival analysis was then conducted to determine its role in the acid stress response. Deletion of yfdX increased the sensitivity of K. pneumoniae CG43S3 to a pH of 2.5, and transforming the mutant with a plasmid carrying yfdX restored the acid resistance (AR) levels. In addition, the effect of yfdX deletion was cross-complemented by the expression of the periplasmic chaperone HdeA. Furthermore, the purified recombinant protein YfdX reduced the acid-induced protein aggregation, suggesting that YfdX as well as HdeA functions as a chaperone. The following promoter activity measurement revealed that rcsB deletion reduced the expression of yfdX after the bacteria were subjected to pH 4.4 adaptation. Western blot analysis also revealed that YfdX production was inhibited by rcsB deletion and only the plasmid expressing RcsB or the nonphosphorylated form of RcsB, RcsBD56A, could restore the YfdX production, and the RcsB-mediated complementation was no longer observed when the sensor kinase RcsD gene was deleted. In conclusion, this is the first study demonstrating that YfdX may be involved in the acid stress response as a periplasmic chaperone and that RcsB positively regulates the acid stress response partly through activation of yfdX expression. Moreover, the phosphorylation status of RcsB may affect the YfdX expression under acidic conditions.

The mass burial of dinosaur bone fossils in the Late Cretaceous Wangshi Group in Zhucheng, Shandong Province has been a research focus in recent years. However, the provenance of the dinosaur bone fossils and the accurate depositional age of the bone beds remain ambiguous. Through U–Pb dating of detrital zircons collected from six conglomerate samples from the dinosaur bone beds, we found that the youngest single grain age (YSG) of sample 090414-24-D was 77.3 Ma, representing the maximum depositional age of the dinosaur fossil beds and sediments. This also indicates that the Hongtuya Formation was deposited during the Campanian. Dating results revealed an age peak of 120–110 Ma, which corresponds with the peak age of volcanic rocks of the Lower Cretaceous Qingshan Group. The volcanic rocks of the Qingshan Group are mainly exposed in Laiyang, to the north of Zhucheng, although a few also appear to the south and northwest. Through analysis of conglomerate composition and palaeocurrents in the sediments containing the bone beds, we found that the three different data sets of gravel compositions of the conglomerates were mainly composed of volcanic or pyroclastic rocks. Three different data sets of palaeocurrents suggested that the main sediment source of the Wangshi Group dinosaur bone beds was from the north−northwest of the Basin. Only one data set had a provenance south of the basin. This study revealed that the areas of Laiyang and the Yishu Fault Zone were the main provenance areas of both the dinosaur bone fossils and the sediments of the Wangshi Group in Zhucheng. The southern margin of the Zhucheng Basin may be a secondary source area. This research provides an important basis for judging the deposition time and the sediment source of fossil layers in the Wangshi Group, as well as reconstructing the palaeogeography of the Wangshi Group in the Jiaolai Basin.

Saccharomyces boulardii preparations are promising probiotics and clinical agents for animals and humans. This work focused on optimizing the nutritional conditions for the production of S. boulardii in solid-state fermentation by using classical and statistical methods. In single-factor experiments, the S. boulardii production was significantly increased by the addition of glucoamylase and the optimal carbon and nitrogen sources were found to be soluble starch and NH4Cl, respectively. The effects of the glucoamylase, soluble starch and NH4Cl on S. boulardii production were evaluated by a three-level three-factor Box–Behnken design and response surface methodology (RSM). The maximal yeast count (4.50 ×109CFU/g) was obtained under the optimized conditions (198 U/g glucoamylase, 2.37% soluble starch and 0.9% NH4Cl), which was in a good agreement with the predicted value of the model. This study has provided useful information on how to improve the accumulation of yeast cells by RSM.

Antibiotic resistance is becoming the biggest threat to global health. At the same time, phage therapy is witnessing a return of interest. The therapeutic use of bacteriophages that infect and kill bacteria is a suitable strategy to combat antibiotic resistance. Furthermore, bacteriophages are increasingly used in combination with standard antibiotics against drug-resistant pathogens. Interestingly, we found that the engineered mycobacteriophage phAE159 and natural phage D29 cannot infect the Mycobacterium tuberculosis in the presence of kanamycin, hygromycin or streptomycin, but the phage infection was not affected in the presence of spectinomycin. Based on a series of studies and structural analysis of the above four aminoglycoside antibiotics, it could be speculated that the amino sugar group of aminoglycoside might selectively inhibit mycobacteriophage DNA replication. Our discovery that broad-spectrum antibiotics inhibit phage infection is of great value. This study will provide guidance for people to combine phage and antibiotics to treat M. tuberculosis.

Antibiotic-resistant bacteria infections pose a threat to public health. Considering the difficulty in developing new antibiotics, it is an urgent need to develop alternative therapies against bacterial pathogens. Bacteriophages (phages) are evaluated as potential substitutes or adjuncts of antibiotics because they are abundant in nature and could specifically lyse bacteria. In this review, we briefly introduce phage therapy and its advantages compared with traditional antibiotic therapy. We also summarize new emerging phage technologies, such as CRISPR-Cas, synthetic phages, etc., and discuss some possible obstacles and potential risks in the application process. We believe that, with the advancement in synthetic biology and delivery technology, phage therapy has broad prospects in the future.

Red swamp crayfish (Procambarus clarkii) breeding is an important economic mainstay in Hubei province, China. However, information on the gut microbiota of the red swamp crayfish is limited. To address this issue, the effect of developmental stage, diet (fermented or non-fermented feed), and geographical location on the gut microbiota composition in the crayfish was studied via high-throughput 16S rRNA gene sequencing. The results revealed that the dominant phyla in the gut of the crayfish were Proteobacteria, Bacteroidetes,Firmicutes, Tenericutes, and RsaHF231. The alpha diversity showed a declining trend during development, and a highly comparable gut microbiota clustering was identified in a development-dependent manner. The results also revealed that development, followed by diet, is a better key driver for crayfish gut microbiota patterns than geographical location. Notably, the relative abundance of Bacteroidetes was significantly higher in the gut of the crayfish fed with fermented feed than those fed with non-fermented feed, suggesting the fermented feed can be important for the functions (e.g., polysaccharide degradation) of the gut microbiota. In summary, our results revealed the factors shaping gut microbiota of the crayfish and the importance of the fermented feed in crayfish breeding.

Lignocellulose comprised of cellulose and hemicellulose is one of the most abundant renewable feedstocks. Lactic acid bacteria have the ability to ferment sugar derived from lignocellulose. In this study, Pediococcus acidilactici PA204 is a lactic acid bacterium with a high tolerance of temperature and high-efficiency utilization of xylose. We developed a fed-batch simultaneous saccharification and fermentation (SSF) process at 37 °C (pH 6.0) using the 30 FPU (filter paper units)/g cellulase and 20 g/L corn steep powder in a 5 L bioreactor to produce lactic acid (LA). The titer, yield, and productivity of LA produced from 12% (w/w) NaOH-pretreated and washed stover were 92.01 g/L, 0.77 g/g stover, and 1.28 g/L/h, respectively, and those from 15% NaOH-pretreated and washed stover were 104.11 g/L, 0.69 g/g stover, and 1.24 g/L/h, respectively. This study develops a feasible fed-batch SSF process for LA production from corn stover and provides a promising candidate strain for high-titer and -yield lignocellulose-derived LA production.

Accurately locating faults is quite important, especially when the geographical environment is complicated. If the exact location of the fault is not given, wrong route would be chosen, which will greatly slow down repair. This paper proposes an improved traveling wave method by compensating the electrical distance of transmission lines. The catenary model is constructed that considers parameters of the tower and the actual temperature. The actual line length is also derived by the catenary model. A 500 kV transmission line model is established by PSCAD/EMTDC. Various fault simulations are conducted and the results demonstrate that the presented method effectively reduces the error ratio of faulty segment positioning and locates faults with high accuracy.

Arid and semi-arid regions comprise nearly one-fifth of the earth's terrestrial surface. However, the diversities and functions of their soil microbial communities are not well understood, despite microbial ecological importance in driving biogeochemical cycling. Here, we analyzed the geochemistry and microbial communities of the desert soils from Tarim Basin, northwestern China. Our geochemical data indicated half of these soils are saline. Metagenomic analysis showed that bacterial phylotypes (89.72% on average) dominated the community, with relatively small proportions of Archaea (7.36%) and Eukaryota (2.21%). Proteobacteria, Firmicutes, Actinobacteria, and Euryarchaeota were most abundant based on metagenomic data, whereas genes attributed to Proteobacteria, Actinobacteria, Euryarchaeota, and Thaumarchaeota most actively transcribed. The most abundant phylotypes (Halobacterium, Halomonas, Burkholderia, Lactococcus, Clavibacter, Cellulomonas, Actinomycetospora, Beutenbergia, Pseudomonas, and Marinobacter) in each soil sample, based on metagenomic data, contributed marginally to the population of all microbial communities, whereas the putative halophiles, which contributed the most abundant transcripts, were in the majority of the active microbial population and is consistent with the soil salinity. Sample correlation analyses according to the detected and active genotypes showed significant differences, indicating high diversity of microbial communities among the Tarim soil samples. Regarding ecological functions based on the metatranscriptomic data, transcription of genes involved in various steps of nitrogen cycling, as well as carbon fixation, were observed in the tested soil samples. Metatranscriptomic data also indicated that Thaumarchaeota are crucial for ammonia oxidation and Proteobacteria play the most important role in other steps of nitrogen cycle. The reductive TCA pathway and dicarboxylate-hydroxybutyrate cycle attributed to Proteobacteria and Crenarchaeota, respectively, were highly represented in carbon fixation. Our study reveals that the microbial communities could provide carbon and nitrogen nutrients for higher plants in the sandy saline soils of Tarim Basin.

In North China, the Mesozoic terrestrial basins, sedimentary palaeogeography and tectonic settings involved five evolutionary stages: (1) the Early‒Middle Triassic, (2) the Late Triassic to Early‒Middle Jurassic, (3) the Late Jurassic to early Early Cretaceous, (4) the middle‒late Early Cretaceous and (5) the Late Cretaceous. The regional punctuated tectonic events occurred during these evolutionary stages. During the Early‒Middle Triassic (stage 1), the Xingmeng Orogenic Belt (XMOB, i.e., eastern part of Central Asia Orogenic Belt, CAOB) of the northern North China was settled in the transition of tectonic environment from syn‒orogenic compression to post‒orogenic extension with intensive uplifting. It is a main provenance in the unified Ordos‒North China Basin.The united continental plate of China and the Qinling‒Dabie‒Sulu Orogenic Belt formed due to convergence and collision between the North China Plate and the Yangtze Plate along two suture zones of the Mianlue and the Shangdan in the terminal Middle Triassic. During the Late Triassic to the Early‒Middle Jurassic (stage 2), the Late Triassic mafic or alkaline rocks and intrusions occurred on the northern and southern margins of North China Craton (NCC) and XMOB, implying that intensified extension happened all over the North China (early phase of stage 2). Additionally, in the late phase of stage 2, the basic volcanicfilling faulted basins were widely distributed in the northeastern North China during the Early–Middle Jurassic, including a series of small- to medium-sized basins with coal-bearing strata and some volcanic rocks in other areas of North China, which was the result of subduction of the Palaeo-Pacific Plate during the Early–Middle Jurassic. An active continental margin with accretionary complex developed in the eastern Heilongjiang of China, Japan and the Far East of Russia at that time. However, in the end of the Early‒Middle Jurassic, because of the Yanshanian orogeny characterized by complicated thrust and fold, the previous unified Ordos–North China Basin was separated by the northeast-oriented Great Xing’an Range and Taihang Mountain uplifted linearment. The differential evolution of basins and sedimentary palaeogeography between eastern and western North China was initiated, and was interpreted to result in the closure of Okhotsk Ocean and the subduction of Palaeo-Pacific Plate (late stage 2). During the Late Jurassic (the early phase of stage 3), a variety of faulted basins occurred in the Yanshan and Yinshan areas in the northeastern North China. In Yanshan area, basins were filled with thickened intermediate volcanic rocks and purple-red coarse-grained clastic rocks. In contrast, only thick layered sedimentary rocks with rare volcanic rocks developed in the Yinshan faulted basins, the Ordos Basin and basins in sourthern North China. XMOB was the main provenance of the Early Mesozoic basins in the North China, while the Ordos Basin and the Hefei Basin were partly supplied by the northern Qinling Orogenic Belt. During the Late Jurassic–early Early Cretaceous (the late phase of stage 3), the northern and northeastern North China experienced extensional movement after the subduction of the Palaeo-Pacific Plate, the closure of the Mongolia–Okhotsk Ocean and the subsequent Yanshanian orogeny. At the same time, a NE-oriented, giant rift basin system (NE Asia Rift) extended from the Yanshan to the western Great Xing’an Range, where rift basins were filled with the regional, NE-oriented, thick coarse-grained clastic rocks and a belt of volcanic rocks. In the meantime, the eastern and northeastern China and most areas of NCC were presented as highland terrains. During the middle‒late Early Cretaceous (stage 4), rift basins developed and accumulated alluvial sediments and interbedded alkaline volcanic rocks in the western and northern North China, including Yingen, Ejinaqi and Erlian regions. Basins were formed on both sides of the Tan-Lu Fault Zone under a striking-slipping force. Furthermore, faulted basins developed in the Yishu Fault Zone of Shandong (central Tan-Lu Fault Zone) as well, where dinosaur fauna flourished. Basic volcanic rocks and fluvial‒lacustrine sediments were deposited in small- or medium-sized rift basins in the northeastern China. The Songliao Basin was a typical giant basin that was mainly filled with late Early Cretaceous lacustrine sediments. A group of rift basins occurred in the Sanjiang area, central Heilongjiang Province, northeastern China. From the middle‒late Early Cretaceous to the Late Cretaceous (stage 5), depositional and subsiding center of the basins constantly shifted southeastwards in Heilongjiang Province. The tectonic setting changed into the Palaeo-Pacific continental margin in north and northeastern China. Besides, during the Late Mesozoic, a huge terrestrial biota, mainly dinosaur fauna, dominated in North China. The Yanliao biota of the Middle–Late Jurassic and the Jehol biota of the Early Cretaceous are characterized by feathered dinosaurs, primitive birds, mammals, pterosaur, insects and plants (angiosperms). In northeastern Asia, this Late Mesozoic tectonic background , palaeogeoraphy and palaeoecology were shared by East China, Korean Peninsula, Japan and the Far East of Russia.

The key technique of a kinetic energy rod (KER) warhead is to control the flight attitude of rods. The rods are usually designed to different shapes. A new conceptual KER named profiled rod which has large L/D ratio is described in this paper. The elastic dynamic equations of this profiled rod flying at high velocity after detonation are set up on the basis of Euler-Bernoulli beam, and the aeroelastic deformation of profiled rod is calculated by semi-analytical method for calculating the vibration characteristics of variable cross-section beam. In addition, the aeroelastic deformation of the undeformed profiled rod and the aeroelastic deformation of deformed profiled rod which is caused by the detonation of explosive are simulated by computational fluid dynamic and finite element method (CFD/FEM), respectively. A satisfactory agreement of these two methods is obtained by the comparison of two methods. The results show that the semi-analytical method for calculating the vibration characteristics of variable cross-section beam is applied to analyze the aeroelastic deformation of profiled rod flying at high velocity.

The aim of this study is to investigate the diagnostic efficacy of neutrophil-to-lymphocyte ratio (NLR), neutrophil-to-monocyte ratio (NMR), lymphocyte-to-monocyte ratio (LMR), and platelet-to-lymphocyte ratio (PLR) in patients with Crohn’s disease (CD) and non-CD controls. These ratios were all derived from complete blood counts. Two hundred and six participants including CD inpatients and non-CD controls were retrospectively enrolled. We found statistically higher NLR and PLR and lower LMR in CD patients than in non-CD controls (all P

This research focused on the effects of different doses of Bacillus subtilis KN-42 on the growth performance, diarrhea incidence, faecal bacterial flora, and the relative number of Lactobacillus and Escherichia coli in faeces of weaned piglets to determine whether the strain can serve as a candidate antimicrobial growth promoter. A total of 360 piglets (initial body weight 7.14±0.63 kg) weaned at 26±2 days of age were randomly allotted to 5 treatment groups (4 pens per treatment with 18 pigs per pen) for a 28-day trial. Dietary treatments were basal diet without any antimicrobial (negative control; NC), basal diet supplemented with 120 mg/kg feed of neomycin sulfate (positive control; PC) and basal diet supplemented with 2×109 (L), 4×109 (M) and 20×109 (H) CFU/kg feed of B. subtilis KN-42. During the overall period, average daily gain and feed efficiency of piglets were higher in groups PC, M, and H than those in group NC (p

This paper proposes an optimal placement model of ultra-high frequency (UHF) sensors for accurate location of partial discharge (PD) in gas-insulated switchgear (GIS). The model is based on 0-1 program in consideration of the attenuation influence on the propagation of electromagnetic (EM) waves generated by PD in GIS. the optimal placement plan improves the economy, observability, and accuracy of PD locating. After synchronously acquiring the time of the initial EM waves reaching each UHF sensor, PD occurring time can be obtained. Then, initial locating results can be acquired by using the Euclidean distance measuring method and the extended time difference of arriving (TDOA) location method. With the information of all UHF sensors and the inherent topological structure of GIS, the locating accuracy can be further improved. The method is verified by experiment, showing that the method can avoid the influence of false information and obtain higher locating accuracy by revising initial locating results.