Petershemmingsen4283

The rise of bacterial antibiotic resistance coupled with a diminished antibiotic drug pipeline underlines the importance of developing rational strategies to discover new antimicrobials. Microbially derived natural products are the basis for most of the antibiotic arsenal available to modern medicine. Here, Luminespib demonstrate a resistance-based approach to identify producers of elfamycins, an under-explored class of natural product antibiotics that target the essential translation factor EF-Tu. Antibiotic producers carry self-resistance genes to avoid suicide. These genes are often found within the same biosynthetic gene cluster (BGC) responsible for making the antibiotic, and we exploited this trait to identify members of the kirromycin class of elfamycin producers. Genome mining of Streptomyces spp. led to the identification of three isolates that harbor kirromycin-resistant EF-Tu (EF-TuKirR) within predicted natural product BGCs. Activity-guided purification on extracts of one of the Streptomyces isolates, which was not known to produce an elfamycin, identified it as a producer of phenelfamycin B, a linear polyketide. Phenelfamycin B demonstrates impressive antibacterial activity (MIC ∼ 1 μg/mL) against multidrug-resistant Neisseria gonorrhoeae, a clinically important Gram negative pathogen. The antigonococcal activity of phenelfamycin was shown to be the result of inhibition of protein biosynthesis by binding to EF-Tu. These results indicate that a resistance-based approach of identifying elfamycin producers is translatable to other antibiotic classes that can identify new and overlooked antibiotics necessary to address the antibiotic crisis.Acute lung injury (ALI)/Acute respiratory distress syndrome (ARDS) is a very dangerous disease. The purpose of this study was to investigate the effects of fibrogrowth factor-2 (FGF-2) on lipopolysaccharide (LPS)-induced lung injury and its mechanisms. C57/BL6 mice were used in the study and LPS was used to construct the ALI/ARDS model. In addition, human normal lung epithelial cell line BEAS-2B was cultured to investigate the effect of FGF-2 on the lung and its mechanism of action in vitro. FGF-2 significantly reduced wet/dry weight ratio of mice, the number of cells and inflammatory factors in BALF, and MPO activity in lung tissue. In addition, FGF-2 also reduced the level of oxidative stress in mouse lung tissue. In vitro, FGF-2 effectively reduced LPS-induced inflammatory and apoptotic levels of BEAS-2B cells and increased the activity of the PI3K/Akt signaling pathway. However, LY294002, an inhibitor of the PI3K/Akt signaling pathway, alleviated the protective effect of FGF-2 on lung tissue. Therefore, FGF-2 attenuated inflammation, oxidative stress and apoptosis in alveolar epithelial cells by activating the PI3K/Akt signaling pathway.Traumatic brain injury (TBI) is a risk factor for the later development of dementia, but although the evidence dates back to the early 20th century, the nature of any association and its mechanistic pathways remain unclear. There has been greater focus on this subject over recent years, in part because of increasing reports around sports related TBIs, especially in the USA. Differences in research methods and clinical sampling remain the primary reason for the variable findings, although there is clearly increased prevalence of neurodegenerative disorders in general. Duration of follow up, definition of both TBI and dementia, and differences in the extent to which other dementia risk factors are controlled, as well as concerns about medical record accuracy are all issues yet to be resolved in TBI research, as is an absence pathological evidence. In addition, TBI has been reported to initiate a cascade of pathological processes related to several neurodegenerative disorders, and as such, it is likely that the risks vary between individuals. Given the evidence that dementia risk may increase with injury severity and frequency, a detailed account of age and type of injury, as well as lifetime TBI exposure is essential to document in future studies, and further longitudinal research with biomarker assessments are needed.While the NHS aims to respect the human rights of every individual, it also has a wider social duty to promote equality in the services it provides. This means that the rights of individual patients are not absolute, because the aim of the NHS is to improve the overall health and well-being of the nation. For example, certain treatments may be withheld from individuals because of the excessive cost to the NHS, or concerns about its clinical effectiveness. This article explains the origins of human rights and their function, and examines the relationship between nursing care and human rights.Technical English is a compulsory course for brewing engineering undergraduates. To improve the quality and effectiveness of teaching, we introduced science, technology, engineering, arts and mathematics (abbreviated to STEAM) education theory in technical English teaching of brewing engineering students, and selected, designed, integrated and optimized the teaching contents. With the help of the Rain Classroom, Massive Open Online Courses of Tsinghua University (XuetangX), Open Language, Wine Folly, training courses of Wine and Spirit Education Trust (WSET) and other platform resources, teaching methods, assessment ways and multi-element teaching evaluation system have been innovatively reformed. By using advanced teaching methods such as mind mapping drawing, micro-video production, situational dialogues, topic speech, creative program design and design competition of wine label and bottle packaging, it could effectively improve students' learning interest and engagement, and enhance students' knowledge of professional English and comprehensive application ability.New engineering program requires training models that conform acceptable time span and principles of engineering education. #link# Considering the program "Principles of chemical engineering" and the limitations of traditional teaching methods, we integrated problem-based learning method and flipped classroom teaching model to reform the course. Through a three-stage systematic teaching design including knowledge learning before class, knowledge internalization in class, and consolidation and expansion after class, we effectively stimulated students' interest and enthusiasm in learning, cultivated students' independent learning ability and engineering thinking, and achieved good teaching effect. It can provide reference for the construction of "Principles of chemical engineering" course and training of engineering talents in agricultural colleges.ω-transaminase (ω-TA) is the most promising biocatalyst for chiral amine synthesis. However, most wild-type ω-TAs cannot be applied in industry directly due to their low stability and unfavorable reaction equilibrium. In order to discover a novel ω-TA for industrial application, we designed a procedure of adaptive selection, including the screening of substrates, protein sequences and clones, enzyme activity, and product conversion and characterization, as well as trouble-shooting of each step. Through this procedure, we screened a novel ω-TA, ATA-W12 of Caulobacter sp. from a soil metagenome. The strain could convert 20 mmol/L 1-Boc-3-pyrrolidinone and 20 mmol/L 1-Boc-3-piperidone with 85.84% and 67.42% conversion rate, respectively, in a 1-mL scale with isopropylamine (IPA) as amine donor. ATA-W12 maintained 100% activity at 40 °C for 168 h, and its optimal reaction condition is at pH 8.5 and 40 °C. These excellent properties benefit the application of IPA as an ideal amino donor in industry. We scaled up the production of (S)-(+)-1-boc-3-aminopiperidine up to 50 mL (100 g/L) scale with this novel biocatalyst for its further industrial application.2-O-α-D-glu-copyranosyl-sn-glycerol is a high value-added product with prospective application in food, cosmetics, health products and pharmaceutical industries. However, industrial scale of 2-O-α-D-glu-copyranosyl-sn-glycerol has not yet been applied in China, and there are few related reports on 2-O-α-D-glu-copyranosyl-sn-glycerol synthesis. The purpose of this experiment is to develop a method for catalyzing the synthesis of food-grade 2-O-α-D-glu-copyranosyl-sn-glycerol using whole cells of "Generally Recognized as Safe" (GRAS) recombinant Bacillus subtilis. In our work, a recombinant B. subtilis 168/pMA5-gtfA that heterologously expressing Leuconostoc mesenteroides sucrose phosphorylase was constructed and used as a whole-cell catalyst to synthesize 2-O-α-D-glu-copyranosyl-sn-glycerol. Optimizing the culture temperature, time and whole cell transformation conditions has increased the yield of 2-O-α-D-glu-copyranosyl-sn-glycerol. The results showed that 1.43 U/mL of sucrose phosphorylase was achieved in B. subtilis 168/pMA5-gtfA after culturing for 20 h at 30 °C in fermentation medium. The highest conversion rate reached 75.1%, and the yield of 2-O-α-D-glu-copyranosyl-sn-glycerol was 189.3 g/L with an average transformation rate of 15.6 mmol/(L·h) after 48 hours whole-cell transformation with the sucrose concentration of 1 mol/L and the glycerol concentration of 2.5 mol/L at 30 °C, OD₆₀₀ 40 and pH 7.0. This is the highest yield of 2-O-α-D-glu-copyranosyl-sn-glycerol synthesized catalytically by recombinant B. subtilis that was ever reported, and this study provides the theoretical and experimental basis for the industrial production and application of 2-O-α-D-glucopyranosyl-sn-glycerol.Glucose biosensor is currently the most common electrochemical biosensor. Most glucose biosensors are prepared by modifying glucose oxidase on the electrode surface. link2 However, in the process of electrode immobilization, enzyme purification is required, which increases the cost and has become a bottleneck in the field of development of immobilized enzyme electrodes. In this study, glucose oxidase (GOD) was displayed on the surface of Bacillus subtilis using the spore capsid protein CotX as an anchor protein. By Western blotting analysis, immunofluorescence analysis and enzyme activity detection, GOD was effectively expressed on the surface of spores, and recombinant spores (Spore-GOD) were obtained by fermentation. The graphene oxide/prussian blue deposition film modified glassy carbon electrode was prepared by the drop coating method and the electrodeposition method. The surface of the modified electrode was fixed with Spore-GOD, and finally covered with a layer of Nafion solution to make an electrochemical biosensor for sensitive determination of glucose. The cyclic voltammogram of glucose on the enzyme electrode sensor showed a well-defined oxidation peak at 0.42 V, and the redox peak current has a good linear relationship with the glucose concentration in the range of 0.1-7.0 mmol/L. The calibration curve equation is I=1.305C(glucose)+3.639 (R²=0.992 9), and its detection limit is 7.5 μmol/L (S/N=3). This modified electrode has good conductivity, stability and reproducibility, and can be used for the analysis and determination of glucose.The main purpose of this research is to synthesize and evaluate a new glycoconjugate vaccine against Klebsiella pneumonia (Kp). First, the gene (waaL) responsible for the expression of O antigen ligase was deleted to block the synthesis of bacterial LPS. Then the vector that encodes a glycosyltransferase (PglL) was transferred into the mutant. link3 The enzyme PglL could catalyze the transfer of OPS units to recombinant cholera toxin B subunit (rCTB) to form glycoproteins in vivo. The protective effects of the glycoproteins were studied by the mice models with acute bacteremia that were induced by intraperitoneal injection of wildtype Kp bacteria. The results were as followings A Kp waaL mutant was obtained and the rCTB protein could be successfully glycosylated in the mutant. The vaccine can stimulate a high antibody titer in the mice sera with or without adjuvant. It can also protect mice from the lethal dose injection of Kp. The survival rate of vaccine candidate groups could reach 75%. The glycoconjugate vaccine candidate prepared by this biosynthetic method is expected to become a novel effective vaccine against Klebsiella pneumoniae.