Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism

BACKGROUND: Although understanding of physiological interactions between plasmid DNA and its host is important for vector design and host optimization in many biotechnological applications, to our knowledge, global studies on plasmid-host interactions have not been performed to date even for well-characterized plasmids. RESULTS: Escherichia coli cells, either devoid of plasmid DNA or bearing plasmid pOri1 (with a single ColE1 replication origin) or plasmid pOri2 (with double ColE1 replication origins), were cultured in a chemostat. We used a combination of metabolic flux analysis, DNA microarray and enzyme activity analysis methods to explore differences in the metabolism between these strains. We found that the presence of plasmids significantly influenced various metabolic pathways in the host cells, e.g. glycolysis, the tricarboxylic acid (TCA) cycle and the pentose phosphate (PP) pathway. Expression of rpiA, a gene coding for ribose-5-phosphate isomerase A, was considerably decreased in E. coli carrying a high copy number plasmid relative to E. coli carrying a low copy number plasmid and plasmid-free E. coli. The rpiA gene was cloned into an expression vector to construct plasmid pETrpiA. Following induction of pETrpiA-bearing E. coli, which harbored either pOri1 or pOri2, with isopropyl-β-D-thiogalactopyranoside (IPTG), the copy number of pOri1 and pOri2 was sigificantly higher than that measured in a host devoid of pETrpiA. CONCLUSION: The presence of plasmids can significantly influence some metabolic pathways in the host cell. We believe that the results of detailed metabolic analysis may be useful in optimizing host strains, vectors and cultivation conditions for various biotechnological purposes

Quantitative Analysis of Water Injection Mass and Timing Effects on Oxy-Fuel Combustion Characteristics in a GDI Engine Fuelled with E10

The climate change issue has become a growing concern due to the increasing greenhouse gas emissions. To achieve carbon neutrality for mitigating the climate problem, the oxy-fuel combustion (OFC) technique on internal combustion engines (ICEs) has attracted much attention. Furthermore, the water injection (WI) strategy was proven effective in improving the combustion process and thermal efficiency in engines under OFC mode. However, WI strategy effects on gasoline direct injection (GDI) engines fuelled with gasoline–alcohol blends have not been reported. This study quantitatively analysed WI mass and timing effects on oxy-fuel combustion performance from a GDI engine fuelled with E10 (10% ethanol and 90% gasoline in mass) by simulation. The results show that equivalent brake-specific fuel consumption (BSFCE) shows a monotonically decreasing trend with the increase in the water–fuel mass ratio

A Numerical Study of the Effects of Oxy-Fuel Combustion under Homogeneous Charge Compression Ignition Regime

The European Union (EU) has recently adopted new directives to reduce the level of pollutant emissions from non-road mobile machinery engines. The main scope of project RIVER for which this study is relating is to develop possible solutions to achieve nitrogen-free combustion and zero-carbon emissions in diesel engines. RIVER aims to apply an oxy-fuel combustion with Carbon Capture and Storage (CCS) technology to eliminate NOx emission and to capture and store carbon emissions. As part of this project, a computational fluid dynamic (CFD) analysis has been performed to investigate the effects of oxy-fuel combustion on combustion characteristics and engine operating conditions in a diesel engine under Homogenous Charge Compression Ignition (HCCI) mode. A reduced chemical n-heptane-n-butanol-PAH mechanism which consists of 76 species and 349 reactions has been applied for oxy-fuel HCCI combustion modeling. Different diluent strategies based on the volume fraction of oxygen and a diluent gas has been considered over a wide range of air-fuel equivalence ratios. Variation in the diluent ratio has been achieved by adding different percentages of carbon dioxide for a range from 77 to 83 vol. % in the intake charge. Results show that indicated thermal efficiency (ITE) has reduced from 32.7% to 20.9% as the CO2 concentration has increased from 77% to 83% at low engine loads while it doesn’t bring any remarkable change at high engine loads. It has also found that this technology has brought CO and PM emissions to very ultra-low level (near zero) while NOx emissions have been completely eliminated

Left Anterior Fascicular Block After Transcatheter Closure of Ventricular Septal Defect in Children

Background: Arrhythmia is the most common complication after transcatheter closure of a ventricular septal defect (VSD). However, the effects of postprocedural left anterior fascicular block are not clear. This study presents the clinical characteristics, prognosis, and related risk factors of left anterior fascicular block after transcatheter closure of a VSD in children.Methods: The clinical and follow-up data of the patients in the Heart Center of Children's Hospital of Chongqing Medical University from June 2009 to October 2018 were reviewed. And 30 cases were eligible out of all 1,371 cases.Results: An electrocardiogram showed a left anterior fascicular block within 3 days, and most patients gradually returned to normal within 1–2 years, showing a dynamic change. Left ventricular end-diastolic dimension Z-score ranged from −2 to 2 in all children, and no decrease of left ventricular ejection fraction was found in all children. The high ratio between VSD size and body surface area [p < 0.05, odds ratio (OR) 2.6, 95% CI: 1.136–6.113] and large diameter difference between the occluder size and VSD size (p < 0.05, OR 2.1, 95% CI: 1.036–4.609) were independent risk factors for postprocedural left anterior fascicular block.Conclusions: The incidence of postprocedural left anterior fascicular block is not that low, and the overall prognosis is quite good at the current follow-up stage. No progressive severity has been found, such as complete left bundle branch block, double (triple) bundle branch block, and atrioventricular block, to have an influence on cardiac systolic and diastolic function

The Investigation of NOx Formation and Reduction during O2/CO2 Combustion of Raw Coal and Coal Char

AbstractThis study investigated the behavior of NOx emissions during combustion of pulverized coal char and pulverized raw coal in O2/CO2 environments under nearly isothermal condition in a drop tube furnace. A representative Chinese coal Ling Nancang (LNC) and LNC CO2-char (made in CO2 condition) were studied with four different excess oxygen ratios (α=0.6, 0.8, 1.2, and 1.4). Combustion experiments were performed with an elevated concentration (850ppm) of NO. When α increased from 0.6 to 1.4, the total amount of NOx reduction was increased in both pulverized coal and char combustion. To investigate the effects of coal and char on NOx reburn reactions, LNC coal char and the raw coal combustion experiments were also performed under three high background NO concentrations (400, 850, and 1200ppm) in O2/CO2 atmosphere. During experiments with elevated NO concentrations, there was an increase in net NO reduction; however, there was also a decrease in the calculated NO reduction ratio