Comparison of different strategies to reduce acetate formation in Escherichia coli

E. coli cells produce acetate as an extracellular coproduct of aerobic cultures. Acetate is undesirable because it retards growth and inhibits protein formation. Most process designs or genetic modifications to minimize acetate formation aim at balancing growth rate and oxygen consumption. In this research, three genetic approaches to reduce acetate formation were investigated: (1) direct reduction of the carbon flow to acetate (ackA-pta, poxB knock-out); (2) anticipation on the underlying metabolic and regulatory mechanisms that lead to acetate (constitutive ppc expression mutant); and (3) both (1) and (2). Initially, these mutants were compared to the wild-type E. coli via batch cultures under aerobic conditions. Subsequently, these mutants were further characterized using metabolic flux analysis on continuous cultures. It is concluded that a combination of directly reducing the carbon flow to acetate and anticipating on the underlying metabolic and regulatory mechanism that lead to acetate, is the most promising approach to overcome acetate formation and improve recombinant protein production. These genetic modifications have no significant influence on the metabolism when growing the micro-organisms under steady state at relatively low dilution rates (less than 0.4 h(-1)).     

Descriptions of two new species of Aelurillus Simon, 1884 (Araneae,Salticidae) from the Mediterranean,with the synonymization of A. steliosi Dobroruka, 2002

Two Aelurillus species are described as new, Aelurillus alboclypeus sp. n. (♂♀, from Turkey) and Aelurillus deltshevi sp. n. (♂, from Macedonia, Bulgaria and Azerbaijan). Aelurillus steliosi Dobroruka, 2002 is synonymized with Aelurillus leipoldae (Metzner, 1999). Additional distributions of the closely related species Aelurillus v-insignitus are provided for the region of study. Distributional maps are provided for the five species reported in this paper.     

MR assessment of bile duct size in healthy individuals: comparison with US measurements

The purpose of the study was to determine the difference in extrahepatic bile duct (EBD) size measured by magnetic resonance (MR) compared with those measured by ultrasound (US). Changes of EBD size related to aging were analyzed too. Size of EBD was measured in 76 randomly selected healthy individuals. Three radiologists blinded to the result of other study preformed measurements by US and three different T2 weighted MR sequences. Correlation and linear regression analysis of obtained data were performed. The mean diameter of EBD measured by US was 3.17 mm and by MR was 3.14 mm on thick slab rapid acquisition with relaxation enhancement (TSE), 3.26 mm on thin section single-shot TSE (HASTE) and 3.30 mm on coronal fully rewound gradient echo (True FISP). There was no statistical difference between US and different MR sequences (p < 0.05). A trend of increase of EBD with age (0.0155 mm per year, p = 0.0954) was observed. Size of EBD highly correlated for each MR sequence with US measurement validating use of MR as a reliable method for evaluation of EBD size. This conclusion is stressed by increase of EBD size with age demonstrated by all measuring methods.     

Targeting Caveolin-1 and Claudin-5 with AY9944, Improve Blood–Brain Barrier Permeability; Computational Simulation and Experimental Study

Cellular and Molecular Neurobiology - The current study aimed to determine the protective effect of AY9944 related to Caveolin-1 and Claudin-5 role in lipid raft, which can rescue the...     

Construction and model-based analysis of a promoter library for <Emphasis Type="Italic">E. coli</Emphasis>: an indispensable tool for metabolic engineering

Background  

Nowadays, the focus in metabolic engineering research is shifting from massive overexpression and inactivation of genes towards the model-based fine tuning of gene expression. In this context, the construction of a library of synthetic promoters of Escherichia coli as a useful tool for fine tuning gene expression is discussed here.     

ESAT-6 from Mycobacterium tuberculosis dissociates from its putative chaperone CFP-10 under acidic conditions and exhibits membrane-lysing activity

The 6-kDa early secreted antigenic target ESAT-6 and the 10-kDa culture filtrate protein CFP-10 of Mycobacterium tuberculosis are secreted by the ESX-1 system into the host cell and thereby contribute to pathogenicity. Although different studies performed at the organismal and cellular levels have helped to explain ESX-1-associated phenomena, not much is known about how ESAT-6 and CFP-10 contribute to pathogenesis at the molecular level. In this study we describe the interaction of both proteins with lipid bilayers, using biologically relevant liposomal preparations containing dimyristoylphosphatidylcholine (DMPC), dimyristoylphosphatidylglycerol, and cholesterol. Using flotation gradient centrifugation, we demonstrate that ESAT-6 showed strong association with liposomes, and in particular with preparations containing DMPC and cholesterol, whereas the interaction of CFP-10 with membranes appeared to be weaker and less specific. Most importantly, binding to the biomembranes no longer occurred when the proteins were present as a 1:1 ESAT-6.CFP-10 complex. However, lowering of the pH resulted in dissociation of the protein complex and subsequent protein-liposome interaction. Finally, cryoelectron microscopy revealed that ESAT-6 destabilized and lysed liposomes, whereas CFP-10 did not. In conclusion, we propose that one of the main features of ESAT-6 in the infection process of M. tuberculosis is the interaction with biomembranes that occurs after dissociation from its putative chaperone CFP-10 under acidic conditions typically encountered in the phagosome.     

Investigation of the extent of contamination of heavy metals in agricultural soil using statistical analyses and contamination indices

To examine the level of contamination of Eshghabad region, Neyshabur, the concentration of heavy metals in soil was measured. For this purpose, 37 samples were taken from surface soil and the total concentration of heavy elements was measured by inductively coupled plasma (ICP). The mean concentration of Pb, Ni, Cu, Zn, and Fe was obtained to be 195, 87.3, 22.8, 274.8, and 2.5%, respectively. Using five valid indicators, the intensity of metal contamination in soil was calculated and compared. Furthermore, using statistical analyses, the relationships between the elements, their origins, and the spatial distribution of metals across various stations were investigated. The results indicated that the mean value of all of the studied metals (except for iron and copper) is greater than the mean concentration of metals in the Earth's crust. The indicators showed very high contamination for lead, while low to medium contamination for other elements of interest in the farming soils of the region. Statistical analyses indicated that there is a relatively similar contamination intensity across all of the studied stations. Considering the quality standards, the soils of this region are threatened by contamination of lead.     

The mitochondrial outer membrane protein MDI promotes local protein synthesis and mtDNA replication

Early embryonic development features rapid nuclear DNA replication cycles, but lacks mtDNA replication. To meet the high‐energy demands of embryogenesis, mature oocytes are furnished with vast amounts of mitochondria and mtDNA. However, the cellular machinery driving massive mtDNA replication in ovaries remains unknown. Here, we describe a Drosophila AKAP protein, MDI that recruits a translation stimulator, La‐related protein (Larp), to the mitochondrial outer membrane in ovaries. The MDI‐Larp complex promotes the synthesis of a subset of nuclear‐encoded mitochondrial proteins by cytosolic ribosomes on the mitochondrial surface. MDI‐Larp's targets include mtDNA replication factors, mitochondrial ribosomal proteins, and electron‐transport chain subunits. Lack of MDI abolishes mtDNA replication in ovaries, which leads to mtDNA deficiency in mature eggs. Targeting Larp to the mitochondrial outer membrane independently of MDI restores local protein synthesis and rescues the phenotypes of mdi mutant flies. Our work suggests that a selective translational boost by the MDI‐Larp complex on the outer mitochondrial membrane might be essential for mtDNA replication and mitochondrial biogenesis during oogenesis.