Functions of the Clostridium acetobutylicium FabF and FabZ proteins in unsaturated fatty acid biosynthesis

<p>Abstract</p> <p>Background</p> <p>The original anaerobic unsaturated fatty acid biosynthesis pathway proposed by Goldfine and Bloch was based on in <it>vivo </it>labeling studies in <it>Clostridium butyricum </it>ATCC 6015 (now <it>C. beijerinckii</it>) but to date no dedicated unsaturated fatty acid biosynthetic enzyme has been identified in Clostridia. <it>C. acetobutylicium </it>synthesizes the same species of unsaturated fatty acids as <it>E. coli</it>, but lacks all of the known unsaturated fatty acid synthetic genes identified in <it>E. coli </it>and other bacteria. A possible explanation was that two enzymes of saturated fatty acid synthesis of <it>C. acetobutylicium</it>, FabZ and FabF might also function in the unsaturated arm of the pathway (a FabZ homologue is known to be an unsaturated fatty acid synthetic enzyme in enterococci).</p> <p>Results</p> <p>We report that the FabF homologue located within the fatty acid biosynthetic gene cluster of <it>C. acetobutylicium </it>functions in synthesis of both unsaturated fatty acids and saturated fatty acids. Expression of this protein in <it>E. coli </it>functionally replaced both the FabB and FabF proteins of the host in <it>vivo </it>and replaced <it>E. coli </it>FabB in a defined in <it>vitro </it>fatty acid synthesis system. In contrast the single <it>C. acetobutylicium </it>FabZ homologue, although able to functionally replace <it>E. coli </it>FabZ in <it>vivo </it>and in <it>vitro</it>, was unable to replace FabA, the key dehydratase-isomerase of <it>E. coli </it>unsaturated fatty acid biosynthesis in <it>vivo </it>and lacked isomerase activity in <it>vitro</it>.</p> <p>Conclusion</p> <p>Thus, <it>C. acetobutylicium </it>introduces the double of unsaturated fatty acids by use of a novel and unknown enzyme.</p

The Structural Characterization and Antigenicity of the S Protein of SARS-CoV

The corona-like spikes or peplomers on the surface of the virion under electronic microscope are the most striking features of coronaviruses. The S (spike) protein is the largest structural protein, with 1,255 amino acids, in the viral genome. Its structure can be divided into three regions: a long N-terminal region in the exterior, a characteristic transmembrane (TM) region, and a short C-terminus in the interior of a virion. We detected fifteen substitutions of nucleotides by comparisons with the seventeen published SARS-CoV genome sequences, eight (53.3%) of which are non-synonymous mutations leading to amino acid alternations with predicted physiochemical changes. The possible antigenic determinants of the S protein are predicted, and the result is confirmed by ELISA (enzyme-linked immunosorbent assay) with synthesized peptides. Another profound finding is that three disulfide bonds are defined at the C-terminus with the N-terminus of the E (envelope) protein, based on the typical sequence and positions, thus establishing the structural connection with these two important structural proteins, if confirmed. Phylogenetic analysis reveals several conserved regions that might be potent drug targets

Reversal of Cocaine-Conditioned Place Preference through Methyl Supplementation in Mice: Altering Global DNA Methylation in the Prefrontal Cortex

Analysis of global methylation in cells has revealed correlations between overall DNA methylation status and some biological states. Recent studies suggest that epigenetic regulation through DNA methylation could be responsible for neuroadaptations induced by addictive drugs. However, there is no investigation to determine global DNA methylation status following repeated exposure to addictive drugs. Using mice conditioned place preference (CPP) procedure, we measured global DNA methylation level in the nucleus accumbens (NAc) and the prefrontal cortex (PFC) associated with drug rewarding effects. We found that cocaine-, but not morphine- or food-CPP training decreased global DNA methylation in the PFC. Chronic treatment with methionine, a methyl donor, for 25 consecutive days prior to and during CPP training inhibited the establishment of cocaine, but not morphine or food CPP. We also found that both mRNA and protein level of DNMT (DNA methytransferase) 3b in the PFC were downregulated following the establishment of cocaine CPP, and the downregulation could be reversed by repeated administration of methionine. Our study indicates a crucial role of global PFC DNA hypomethylation in the rewarding effects of cocaine. Reversal of global DNA hypomethylation could significantly attenuate the rewarding effects induced by cocaine. Our results suggest that methionine may have become a potential therapeutic target to treat cocaine addiction

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Constant elasticity of substitution function based RANSAC for image stitching

Feature matching is very important in image stitching. RANSAC algorithm is a representative algorithm for feature matching. However, RANSAC still has many shortcomings such as a large number of iterations, a large computational complexity and cannot completely eliminate mismatches. To address above problem, in this paper, we propose a novel method termed constant elasticity of substitution function based RANSAC (CES-RANSAC) for image stitching. Specifically, CES-RANSAC improves the RANSAC algorithm by constructing a utility function, optimizing the boundary of the utility function, calculating Cobb-Douglas coefficients. It also introduces Lindahl equilibrium to derive the return value t to help eliminate mismatches. Experiments show that compared with the traditional RANSAC algorithm, CES-RANSAC has improved matching accuracy and increased computational efficiency, which further improves the efficiency of the image matching algorithm

Design and Application of a Rock Porosity Measurement Apparatus under High Isostatic Pressure

Rock porosity is a key physical parameter at room temperature and pressure that plays an important role in evaluating reserves of oil and natural gas. Research on rock porosity spans over a hundred years. However, in situ porosity under a high isostatic pressure has not been adequately explored, and the experimental conditions for measuring porosity remain unclear. To investigate the feasibility of porosity measurement under a high isostatic pressure and the optimal choice of experimental conditions for this, we design an experimental apparatus that can achieve isostatic pressure up to 200 MPa to fit the relationship between the void volume of a given sample and the drop in gas pressure in an empty standard chamber. The effect of experimental parameters, such as the initial gas pressure at the inlet, the time needed for the gas to reach equilibrium, and the time needed for vacuuming, on the porosity experiment was examined. A series of porosity experiments under different isostatic pressures of up to 200 MPa were carried out with this apparatus. The results quantitatively verify the degree to which porosity is related to isostatic pressure

Numerical Simulation of Flood Intrusion Process under Malfunction of Flood Retaining Facilities in Complex Subway Stations

In recent years, heavy rain and waterlogging accidents in subway stations have occurred many times around the world. With the comprehensive development trend of underground space, the accidents caused by flood flow intruding complex subway stations and other underground complexes in extreme precipitation disasters will be lead to more serious casualties and property damage. Therefore, it is necessary to conduct numerical simulation of flood intrusion process under malfunction of flood retaining facilities in complex subway stations. In order to prevent floods from intruding subway stations and explore coping strategies, in this study, the simulation method was used to study the entire process of flood intrusion into complex subway stations when the flood retaining facilities fail in extreme rain and flood disasters that occur once-in-a-century. The three-dimensional numerical simulation model was constructed by taking a subway interchange station with a property development floor in Nanning as a prototype. Based on the Volume of Fluid (VOF) model method, the inundated area in the subway station during the process of flood intrusion from the beginning to the basic stability was simulated, and it was found that the property development floor has serious large-scale water accumulation under extreme rainfall conditions. Through the dynamic monitoring of the flood water level depth at important positions such as the entrances of the evacuation passages, and the analysis of the influence of the design structure and location distribution of different passages on the personnel evacuation plan, it was found that the deep water accumulation at the entrances of the narrow, long, and multi-run emergency safety passages are not conducive to the evacuation of personnel. Finally, the flow of flood water into the subway tunnel through the subway station was calculated. The research results provide certain reference and guidance for the safety design of subway stations under extreme rainfall climatic conditions

Numerical Simulation of Flood Intrusion Process under Malfunction of Flood Retaining Facilities in Complex Subway Stations

In recent years, heavy rain and waterlogging accidents in subway stations have occurred many times around the world. With the comprehensive development trend of underground space, the accidents caused by flood flow intruding complex subway stations and other underground complexes in extreme precipitation disasters will be lead to more serious casualties and property damage. Therefore, it is necessary to conduct numerical simulation of flood intrusion process under malfunction of flood retaining facilities in complex subway stations. In order to prevent floods from intruding subway stations and explore coping strategies, in this study, the simulation method was used to study the entire process of flood intrusion into complex subway stations when the flood retaining facilities fail in extreme rain and flood disasters that occur once-in-a-century. The three-dimensional numerical simulation model was constructed by taking a subway interchange station with a property development floor in Nanning as a prototype. Based on the Volume of Fluid (VOF) model method, the inundated area in the subway station during the process of flood intrusion from the beginning to the basic stability was simulated, and it was found that the property development floor has serious large-scale water accumulation under extreme rainfall conditions. Through the dynamic monitoring of the flood water level depth at important positions such as the entrances of the evacuation passages, and the analysis of the influence of the design structure and location distribution of different passages on the personnel evacuation plan, it was found that the deep water accumulation at the entrances of the narrow, long, and multi-run emergency safety passages are not conducive to the evacuation of personnel. Finally, the flow of flood water into the subway tunnel through the subway station was calculated. The research results provide certain reference and guidance for the safety design of subway stations under extreme rainfall climatic conditions

A virtual simulation-based training program on birthing positions: a randomized controlled trial

Abstract Background Restricting parturient women in healthcare facilities from choosing positions that provide the greatest comfort and benefit during labor is a global barrier. Several complex factors, including caregiver preference and medical intervention, shape the limitation. Therefore, a practical need exists to train midwives on the knowledge and skills to change this condition. Methods The study used a parallel, single-blind, randomized controlled trial at a provincial maternity and child health hospital in Fujian, China, from June 1 to December 31, 2019. The midwives in a birth suite were selected and randomly enrolled in a one-month simulation-based hybrid training or face-to-face teaching in September 2019. The four-level Kirkpatrick’s model, including reaction, learning, behavior, and results, was used to evaluate training effects before and after the program. Data were analyzed with SPSS 25.0 using Student’s t-test, Spearman’s correlation test, Mann–Whitney U test, Wilcoxon signed-rank test, and chi-square test analysis of variance. The significance level was set at p 0.05). The virtual simulation group demonstrated an increase in non-supine birth rate (p = 0.030) and a decrease in perineal incision rate among primiparas compared to pre-intervention (p = 0.035). Moreover, knowledge performance was associated with the duration of virtual simulation (r = 0.664, p = 0.001). Conclusions Virtual simulation is a fascinating innovation that enables midwives to develop birthing positions without practicing on real pregnant women and is one solution to achieve work competency within a shortened training period

The impact of grafted surface defects on the on-surface Schiff-base chemistry at the solid-liquid interface

We investigate the effect of covalently modified graphitic surfaces on the formation of single-layer covalent organic frameworks (sCOFs) at the solid-liquid interface. The impact of different densities of grafted aryl species was tested on the on-surface synthesis of three distinct imine-based 2D sCOFs. The grafted aryl species that act as defects provide steric barriers to the progress of the Schiff-base reaction, and can be locally removed to start the 2D polymer healing process. This unique strategy provides a general approach to study in situ this dynamic covalent on-surface chemistry.status: publishe