Computation of steady and unsteady quasi-one-dimensional viscous/inviscid interacting internal flows at subsonic, transonic, and supersonic Mach numbers

Computations of viscous-inviscid interacting internal flowfields are presented for steady and unsteady quasi-one-dimensional (Q1D) test cases. The unsteady Q1D Euler equations are coupled with integral boundary-layer equations for unsteady, two-dimensional (planar or axisymmetric), turbulent flow over impermeable, adiabatic walls. The coupling methodology differs from that used in most techniques reported previously in that the above mentioned equation sets are written as a complete system and solved simultaneously; that is, the coupling is carried out directly through the equations as opposed to coupling the solutions of the different equation sets. Solutions to the coupled system of equations are obtained using both explicit and implicit numerical schemes for steady subsonic, steady transonic, and both steady and unsteady supersonic internal flowfields. Computed solutions are compared with measurements as well as Navier-Stokes and inverse boundary-layer methods. An analysis of the eigenvalues of the coefficient matrix associated with the quasi-linear form of the coupled system of equations indicates the presence of complex eigenvalues for certain flow conditions. It is concluded that although reasonable solutions can be obtained numerically, these complex eigenvalues contribute to the overall difficulty in obtaining numerical solutions to the coupled system of equations

Gene doctoring: a method for recombineering in laboratory and pathogenic Escherichia coli strains

Background: Homologous recombination mediated by the lambda-Red genes is a common method for making chromosomal modifications in Escherichia coli. Several protocols have been developed that differ in the mechanisms by which DNA, carrying regions homologous to the chromosome, are delivered into the cell. A common technique is to electroporate linear DNA fragments into cells. Alternatively, DNA fragments are generated in vivo by digestion of a donor plasmid with a nuclease that does not cleave the host genome. In both cases the lambda-Red gene products recombine homologous regions carried on the linear DNA fragments with the chromosome. We have successfully used both techniques to generate chromosomal mutations in E. coli K-12 strains. However, we have had limited success with these lambda-Red based recombination techniques in pathogenic E. coli strains, which has led us to develop an enhanced protocol for recombineering in such strains. \ud \ud Results: Our goal was to develop a high-throughput recombineering system, primarily for the coupling of genes to epitope tags, which could also be used for deletion of genes in both pathogenic and K-12 E. coli strains. To that end we have designed a series of donor plasmids for use with the lambda-Red recombination system, which when cleaved in vivo by the I-SceI meganuclease generate a discrete linear DNA fragment, allowing for C-terminal tagging of chromosomal genes with a 6xHis, 3xFLAG, 4xProteinA or GFP tag or for the deletion of chromosomal regions. We have enhanced existing protocols and technologies by inclusion of a cassette conferring kanamycin resistance and, crucially, by including the sacB gene on the donor plasmid, so that all but true recombinants are counter-selected on kanamycin and sucrose containing media, thus eliminating the need for extensive screening. This method has the added advantage of limiting the exposure of cells to the potential damaging effects of the lambda-Red system, which can lead to unwanted secondary alterations to the chromosome. \ud \ud Conclusion: We have developed a counter-selective recombineering technique for epitope tagging or for deleting genes in E. coli. We have demonstrated the versatility of the technique by modifying the chromosome of the enterohaemorrhagic O157:H7 (EHEC), uropathogenic CFT073 (UPEC), enteroaggregative O42 (EAEC) and enterotoxigenic H10407 (ETEC) E. coli strains as well as in K-12 laboratory strains

Construction Cost Sensitivity of a Lignocellulosic Ethanol Biorefinery

The technology has been developed to convert feedstock with cellulose content into ethanol. However, ethanol produced from cellulosic feedstock is the same as ethanol distilled from grain. The objective of research is to determine the price per gallon of ethanol needed so that producing lignocellulosic based ethanol become economically feasible.Environmental Economics and Policy, Production Economics,

Activation of σ28-dependent transcription inEscherichia coliby the cyclic AMP receptor protein requires an unusual promoter organization

The Escherichia coli aer regulatory region contains a single promoter that is recognized by RNA polymerase containing the flagellar sigma factor, σ28. Expression from this promoter is dependent on direct activation by the cyclic AMP receptor protein, which binds to a target centred 49.5 base pairs upstream from the transcript start. Activator-dependent transcription from the aer promoter was reconstituted in vitro, and a tethered inorganic nuclease was used to find the position of the C-terminal domains of the RNA polymerase α subunits in transcriptionally competent open complexes. We report that the ternary activator-RNA polymerase-aer promoter open complex is organized differently from complexes at previously characterized promoters. Among other E. coli promoters recognized by RNA polymerase containing σ28, only the trg promoter is activated directly by the cyclic AMP receptor protein. The organization of the different promoter elements and the activator binding site at the trg promoter is the same as at the aer promoter, suggesting a common activation mechanism

Effects of Second Implant on Feedlot Gain and Carcass Traits

Two hundred eighteen steers were finished in a total confinement deep-bedded system at the Armstrong Research Farm, Lewis, IA during 2009. All steers were implanted with Synovex-Choice on day 1 and half the steers in each pen were implanted with Synovex-Choice on day 56. All steers were harvested on day 118. The 2nd implant resulted in an immediate and significant improvement in average daily gain. In the 76 day weigh period following reimplantation the group receiving the 2nd implant gained .66 lb/day more than the group not receiving an additional implant. The overall average daily gain of steers implanted once compared to the steers implanted twice was 3.81 vs. 4.10. The 2nd implant group produced significantly heavier carcasses. There were no significant differences in carcass fat cover or ribeye area. The twice implanted steers had a lower percentage low Choice or better (P=.0571) and a greater percentage Select (P=.0555). Implanting a second time resulted in an increase in carcass weights, an almost significant reduction in % Choice but still resulted in a numerical, non-significant increase in carcass value

DNA Sampling: a method for probing protein binding at specific loci on bacterial chromosomes

We describe a protocol, DNA sampling, for the rapid isolation of specific segments of DNA, together with bound proteins, from Escherichia coli K-12. The DNA to be sampled is generated as a discrete fragment within cells by the yeast I-SceI meganuclease, and is purified using FLAG-tagged LacI repressor and beads carrying anti-FLAG antibody. We illustrate the method by investigating the proteins bound to the colicin K gene regulatory region, either before or after induction of the colicin K gene promoter

The Significance of Finished Cattle Sorting Methodology on Grid Market Performance and Enhanced Revenue for Calf-Fed Beef Cattle

During the past 15 years there has been a major change in the way finished cattle are marketed. Live bids on complete pens of cattle are less prevalent with the advent of value-based marketing where there is an increased emphasis placed on carcass quality and red meat yield. Value-based marketing establishes value based on the animal’s own individual carcass merit. Various grid markets have specifications for important carcass traits that include quality grade, yield grade, and carcass weight. Carcasses that exceed the criteria receive premiums while those that fall short of the specifications receive discounts that in some cases are quite severe. Because of this newer pricing system there may be economic advantages to sort cattle at the end of the feeding period. Past research has demonstrated that sorting cattle by specific traits results in reducing the variation of the traits being evaluated. Feedlots and producers need a sort system that can be performed in a minimal amount of time and expense and is accurate in identifying animals that meet the specifications for a particular market

Early Mesozoic Paleotectonic-Paleogeographic Reconstruction of Southern Sierra Nevada Region

A paleotectonic-paleogeographic reconstruction was based on structural, petrologic, and geochronologic studies of pre-Sierra Nevada batholith framework rocks exposed between the San Joaquin River and the Garlock fault. Most available fossil data from roof pendants of this region indicate Late Triassic to Early Jurassic ages. An additional fossil locality from the western wall rocks yields a Late Permian Tethyan fauna. This is a maximum age for the enclosing rocks, for the fossils are in a limestone olistolith. As yet there is no sign of Paleozoic strata in the region except perhaps along the eastern Sierran crest in small metamorphic septa, and in the western foothills where ophiolitic rocks are present

Constructive Gelfand duality for C*-algebras

We present a constructive proof of Gelfand duality for C*-algebras by reducing the problem to Gelfand duality for real C*-algebras.Comment: 6page

Growing dairy heifers in southwest Iowa

Southwest Iowa farmers were looking for a farming enterprise to add value to their forage and grain production and use their labor. The original plan was to grow dairy heifers on pasture in the summer, sell them in the fall, and keep track of the economics of a dairy heifer system