Enhanced high copy number plasmid maintenance and heterologous protein production in an Escherichia coli biofilm

Escherichia coli has been widely used for heterologous protein production (HPP). To determine whether a biofilm environment could benefit E. coli HPP using high copy number plasmids, we compared plasmid maintenance and HPP by E. coli ATCC 33456 containing plasmid pEGFP (a pUC family vector) cultivated in biofilms and in suspended culture. Cells were grown with or without antibiotic selective pressure in flow cells or chemostats for up to 6 days. In biofilms, antibiotic selective pressure increased the plasmid copy number (PCN), but by 144 h, biofilms grown in antibiotic-free media had comparable plasmid concentrations. In the chemostat, the PCN declined steadily, although 100 ppm ampicillin in the medium slowed the rate of plasmid loss. Production of green fluorescent protein (GFP), a representative heterologous protein, was quantified by flow cytometry. In biofilms, at ampicillin concentrations >or=33 ppm, strongly fluorescent cells comprised more than half of the population by 48 h. In the chemostat, more than 50% of the population was non-fluorescent by 48 h in media containing 100 ppm ampicillin, and strongly fluorescent cells were <10% of the population. Biofilm structure was determined by confocal microscopy. Maximum biofilm thickness ranged from 30 to 45 microns, with no significant changes in biofilm structure after 48 h. Plasmid multimer percentages were similar to inocula for cells cultivated in either biofilms or the chemostat. The results indicate that the biofilm environment enhanced both plasmid maintenance and cellular GFP concentrations, and that low levels of antibiotic increased the beneficial effect.     

Non-genomic effects of aldosterone on intracellular ion regulation and cell volume in rat ventricular myocytes

Aldosterone has non-genomic effects that express within minutes and modulate intracellular ion milieu and cellular function. However, it is still undefined whether aldosterone actually alters intracellular ion concentrations or cellular contractility. To clarify the non-genomic effects of aldosterone, we measured [Na+]i, Ca2+ transient (CaT), and cell volume in dye-loaded rat ventricular myocytes, and we also evaluated myocardial contractility. We found the following: (i) aldosterone increased [Na+]i at the concentrations of 100 nmol/L to 10 micromol/L; (ii) aldosterone (up to 10 micromol/L) did not alter CaT and cell shortening in isolated myocytes, developed tension in papillary muscles, or left ventricular developed pressure in Langendorff-perfused hearts; (iii) aldosterone (100 nmol/L) increased the cell volume from 47.5 +/- 3.6 pL to 49.8 +/- 3.7 pL (n=8, p<0.05); (iv) both the increases in [Na+]i and cell volume were blocked by a Na+-K+-2Cl- co-transporter (NKCCl) inhibitor, bumetanide, or by a Na+/H+ exchange (NHE) inhibitor, 5-(N-ethyl-N-isopropyl) amiloride; and (v) spironolactone by itself increased in [Na+]i and cell volume. In conclusion, aldosterone rapidly increased [Na+]i and cell volume via NKCC1 and NHE, whereas there were no changes in CaT or myocardial contractility. Hence the non-genomic effects of aldosterone may be related to cell swelling rather than the increase in contractility.     

Phylogenetic diversity of bacteria isolates from the Qinghai-Tibet Plateau permafrost region

The Qinghai-Tibet Plateau in east Asia is a unique and important permafrost environment. However, its microbiology remains largely unexplored to date. In this study, sediment samples were collected from the Qinghai-Tibet Plateau permafrost region, bacteria isolation procedures were performed 8 times, and the samples incubated at 4 degrees C for nearly 3 months. The number of colony forming units (cfu) ranged from 0 to 10(7)/(g dry soil). The quantity of culturable bacteria grew exponentially within the first few weeks, and then slowed gradually to a plateau. Phylogenetic analyses indicated that all the isolates fell into 6 categories: high G+C Gram-positive bacteria, low G+C Gram-positive bacteria, alpha-Proteobacteria, beta-Proteobacteria, gamma-Proteobacteria, and Cytophaga-Flavobacterium-Bacteroides group bacteria. The isolates belong to 19 genera, but the genera Arthrobacter and Pseudomonas were predominant. With the increase in incubation time, the isolated populations changed in terms of both species and their respective quantities. Of the 33 analyzed isolates, 9 isolates related to 8 genera might be new taxa. These results suggest that the Qinghai-Tibet Plateau permafrost region is a specific ecologic niche that accommodates an original microbial assemblage.     

Protecting exons from deleterious R-loops: a potential advantage of having introns

Background  

Accumulating evidence indicates that the nascent RNA can invade and pair with one strand of DNA, forming an R-loop structure that threatens the stability of the genome. In addition, the cost and benefit of introns are still in debate.     

Identification of the three subtypes and the prognostic characteristics of stomach adenocarcinoma: analysis of the hypoxia-related long non-coding RNAs

Functional & Integrative Genomics - Stomach adenocarcinoma (STAD) is one of the most commonly diagnosed cancers. This study analyzed the subtypes and characteristics of STAD subtypes by...     

Mechanical analysis of deep tissue injury during sitting in patients with spinal cord injury via parametric finite element model

Biomechanics and Modeling in Mechanobiology - Spinal cord injury patients are prone to develop deep tissue injury because of long-term mechanical load. However, there is a lack of statistical...     

2021年膜翅目新增分类单元

本文对2021年发表的膜翅目昆虫新分类单元进行了梳理和总结。结合数据库检索, 基于标本记录, 全球膜翅目学者于2021年发表该目新分类单元的期刊论文355篇, 新增分类单元条目共1,152条, 隶属于21总科66科416属, 包括5新科4新亚科83新属3新亚属1,054新种和3新亚种。现生类群相关期刊论文309篇, 新增分类单元条目980条, 隶属于18总科52科332属, 包括2新科26新属3新亚属946新种和3新亚种。绝灭类群相关期刊论文46篇, 新增分类单元条目172条, 隶属于14总科27科86属, 包括3新科4新亚科57新属和108新种。2021年中国膜翅目新增分类单元的相关期刊论文83篇, 新增分类单元条目235条, 隶属于17总科34科91属, 包括3新属(绝灭类群1新属)和232新种(绝灭类群2新种); 现生类群中新增的2属分别记录自台湾和浙江, 新种数量排前五位的省级行政单位有云南(54个)、浙江(42个)、福建(18个)、西藏(18个)和新疆(16个)。在全球现生、绝灭和中国现生膜翅目总科新物种数量的对比中, 姬蜂总科新种数量最多, 分别约占全球现生、绝灭和中国现生膜翅目新种总数的32.5% (307个/946个)、19.4% (21个/108个)和37.0% (85个/230个)。有关现生膜翅目新种发表情况, 在洲级地理单元中, 亚洲发表新种数量最多, 约占56.9% (538个); 在洲级地理亚单元中, 东亚发表新种数量最多, 约占28.6% (271个); 在国家和地区行政单元中, 中国发表新种数量最多, 约占24.3% (230个)。在76种期刊的355篇论文中, 有348篇英文论文、4篇中文论文和3篇法语论文。这些结果表明, 中国膜翅目分类在全球膜翅目分类中发挥着十分重要的作用。