The role of Cra in regulating acetate excretion and osmotic tolerance in E. coli K-12 and E. coli B at high density growth

<p>Abstract</p> <p>Background</p> <p><it>E. coli </it>B (BL21), unlike <it>E.coli </it>K-12 (JM109) is insensitive to glucose concentration and, therefore, grows faster and produces less acetate than <it>E. coli </it>K-12, especially when growing to high cell densities at high glucose concentration. By performing genomic analysis, it was demonstrated that the cause of this difference in sensitivity to the glucose concentration is the result of the differences in the central carbon metabolism activity. We hypothesized that the global transcription regulator Cra (FruR) is constitutively expressed in <it>E. coli </it>B and may be responsible for the different behaviour of the two strains. To investigate this possibility and better understand the function of Cra in the two strains, <it>cra </it>- negative <it>E. coli </it>B (BL21) and <it>E. coli </it>K-12 (JM109) were prepared and their growth behaviour and gene expression at high glucose were evaluated using microarray and real-time PCR.</p> <p>Results</p> <p>The deletion of the <it>cra </it>gene in <it>E. coli </it>B (BL21) minimally affected the growth and maximal acetate accumulation, while the deletion of the same gene in <it>E.coli </it>K-12 (JM109) caused the cells to stop growing as soon as acetate concentration reached 6.6 g/L and the media conductivity reached 21 mS/cm. <it>ppsA </it>(gluconeogenesis gene), <it>aceBA </it>(the glyoxylate shunt genes) and <it>poxB </it>(the acetate producing gene) were down-regulated in both strains, while <it>acs </it>(acetate uptake gene) was down-regulated only in <it>E.coli </it>B (BL21). These transcriptional differences had little effect on acetate and pyruvate production. Additionally, it was found that the lower growth of <it>E. coli </it>K-12 (JM109) strain was the result of transcription inhibition of the osmoprotectant producing <it>bet </it>operon (<it>betABT</it>).</p> <p>Conclusions</p> <p>The transcriptional changes caused by the deletion of <it>cra </it>gene did not affect the activity of the central carbon metabolism, suggesting that Cra does not act alone; rather it interacts with other pleiotropic regulators to create a network of metabolic effects. An unexpected outcome of this work is the finding that <it>cra </it>deletion caused transcription inhibition of the <it>bet </it>operon in <it>E. coli </it>K-12 (JM109) but did not affect this operon transcription in <it>E. coli </it>B (BL21). This property, together with the insensitivity to high glucose concentrations, makes this the <it>E. coli </it>B (BL21) strain more resistant to environmental changes.</p

Breakdown of the interlayer coherence in twisted bilayer graphene

Coherent motion of the electrons in the Bloch states is one of the fundamental concepts of the charge conduction in solid state physics. In layered materials, however, such a condition often breaks down for the interlayer conduction, when the interlayer coupling is significantly reduced by e.g. large interlayer separation. We report that complete suppression of coherent conduction is realized even in an atomic length scale of layer separation in twisted bilayer graphene. The interlayer resistivity of twisted bilayer graphene is much higher than the c-axis resistivity of Bernal-stacked graphite, and exhibits strong dependence on temperature as well as on external electric fields. These results suggest that the graphene layers are significantly decoupled by rotation and incoherent conduction is a main transport channel between the layers of twisted bilayer graphene.Comment: 5 pages, 3 figure

EFFECTS OF LIQUID SWIRLING ON GAS-TO-LIQUID MASS TRANSFER IN THREE-PHASE FLUIDIZED BEDS

The swirling flow mode of liquid phase was adopted to promote the gas-to-liquid mass transfer in three-phase(gas-liquid-solid) fluidized beds. Effects of gas(0.01-0.09m/s) and liquid(0.035-0.172m/s) velocities, particle size(1.7-6.0mm) and swirling ratio of liquid phase(0-0.5) on the volumetric gas-to-liquid mass transfer coefficient in the bed were examined. The mass transfer coefficient increased up to 70% by adjusting the swirling flow of liquid phase, especially when the gas velocity is relatively low range. The value of gas-to-liquid mass transfer coefficient was well correlated in terms of dimensionless groups which were derived from the dimensional analysis on the mass transfer system

TTF-1, a homeodomain-containing transcription factor, participates in the control of body fluid homeostasis by regulating angiotensinogen gene transcription in the rat subfornical organ.

In recent years, it has become increasingly evident that angiotensins synthesized in the brain contribute to regulating body fluid homeostasis. Although angiotensinogen, the unique angiotensin precursor, is produced in the brain, the factors that regulate its gene expression remain unknown. We recently found that TTF-1, a homeodomain-containing transcription factor essential for the development of the fetal diencephalon, is postnatally expressed in discrete areas of the hypothalamus. We now report that the subfornical organ, an important site of angiotensinogen synthesis, is an extra-hypothalamic site of TTF-1 expression. Double in situ hybridization histochemistry demonstrated the presence of TTF-1 mRNA in angiotensinogen-producing cells of the rat subfornical organ. RNase protection assays showed that TTF-1 and angiotensinogen mRNA levels are simultaneously increased in the subfornical organ by water deprivation. The angiotensinogen promoter contains seven presumptive TTF-1 binding motifs, four of which are recognized by the TTF-1 homeodomain. In the C6 glioma cell line, TTF-1 transactivates the angiotensinogen promoter in a dose-dependent manner. This transactivation is abolished by deletion of the TTF-1 binding motif at -125. Intracranial administration of an antisense TTF-1 oligodeoxynucleotide decreased angiotensinogen mRNA in the subfornical organ and dramatically reduced the animal's water intake while increasing urine excretion. Moreover, plasma arginine vasopressin content was decreased by the same treatment. These results demonstrate a novel role for TTF-1 in the regulation of body fluid homeostasis, exerted via the transactivational control of angiotensinogen synthesis in the subfornical organ

Molecular characterization of tetracycline- and quinolone-resistant Aeromonas salmonicida isolated in Korea

The antibiotic resistance of 16 Aeromonas (A.) salmonicida strains isolated from diseased fish and environmental samples in Korea from 2006 to 2009 were investigated in this study. Tetracycline or quinolone resistance was observed in eight and 16 of the isolates, respectively, based on the measured minimal inhibitory concentrations. Among the tetracycline-resistant strains, seven of the isolates harbored tetA gene and one isolate harbored tetE gene. Additionally, quinolone-resistance determining regions (QRDRs) consisting of the gyrA and parC genes were amplified and sequenced. Among the quinolone-resistant A. salmonicida strains, 15 harbored point mutations in the gyrA codon 83 which were responsible for the corresponding amino acid substitutions of Ser83→Arg83 or Ser83→Asn83. We detected no point mutations in other QRDRs, such as gyrA codons 87 and 92, and parC codons 80 and 84. Genetic similarity was assessed via pulsed-field gel electrophoresis, and the results indicated high clonality among the Korean antibiotic-resistant strains of A. salmonicida

Late Simultaneous Presentation of Left Ventricular Pseudoaneurysm and Tricuspid Regurgitation after Blunt Chest Trauma

A 32-yr-old man developed progressive exertional dyspnea 4 yr after blunt chest trauma due to an automobile accident. Two-dimensional echocardiography and computed-tomographic coronary angiography demonstrated a large pseudoaneurysm of the left ventricle and severe tricuspid regurgitation. The patient underwent successful surgical exclusion of the pseudoaneurysm by endoaneurysmal patch closure and repair of the tricuspid valve regurgitation. To the best of our knowledge, this is the first case of these 2 different pathologies presenting late simultaneously after blunt chest trauma and successful surgical repairs in the published literature