Metabolic consequences of phosphotransferase (PTS) mutation in a phenylalanine-producing recombinant Escherichia coli

E. coli strain PPA305, which has a wild-type PTS system, and PPA316, which utilizes a proton-galactose symport system for glucose uptake, were used as host strains to harbor a phenylalanine overproduction plasmid pSY130-14 and to study the effects of using different glucose uptake systems on phenylalanine production. The non-PTS strain (PPA316/pSY130-14) produced much less phenylalanine, ranging from 0 to 67% of that produced by the PTS strain (PPA305/pSY130-14) depending on cultivation conditions used. The non-PTS strain PPA316/pSY130-14 had an intracellular PEP concentration only one-sixth that of the PTS strain, PPA305/pSY130-14. Additionally, PPA316/pSY130-14 had a substantially lower energy state in terms of the size of the pool of high-energy phosphate compounds and the magnitude of the pH difference across the cytoplasmic membrane. The non-PTS strain consumed oxygen at a higher rate, attained lower biomass concentration, and produced no acetate and phenylalanine during fermentation, suggesting more carbon was oxidized to CO2, most likely through the TCA cycle. Analysis of intracellular fluxes through the central carbon pathways was performed for each strain utilizing exponential phase data on extracellular components and assuming quasi-steady state for intermediate metabolites. The non-PTS strain had a higher flux through pyruvate kinase (PYK) and TCA cycle which, in agreement with the observed higher oxygen uptake rate, suggests that more carbon was oxidized to CO2 through the TCA cycle. Further analysis using rate expression data for PYK and NMR data for the intracellular metabolites identified the regulatory properties of PYK as the probable cause for lower intracellular PEP levels in PPA316/pSY130-14.     

A new approach to clustering

Estimation theory is used to derive a new approach to the clustering problem. The new method is a unification of centroid and mode estimation, achieved by considering the effect of spatial scale on the estimator. The result is a multiresolution method which spans a range of spatial scales, giving enhanced robustness both to noise in the data and to changes of scale in the data, by using comparison between scales as a test of cluster validity. Iterative and non-iterative algorithms based on the new estimator are presented and are shown to be more accurate than simple scale-space filtering in identifying and locating the cluster centres from noisy test data. Results from a wide range of applications are used to illustrate the power and versatility of the new method.     

Final design of an air core, compulsator driven, 60 caliber railgunsystem

The manufacturing phase of a laboratory-based small-caliber electromagnetic (EM) launcher and compulsator power supply is discussed. The objective of the 29-month program is to develop a compact, lightweight test bed capable of accelerating 32 g masses to 2 km/s at a rate of 10 Hz. Both the power supply and launcher feature significant component design advances which will allow the system to operate at considerably higher energy and power densities than previously demonstrated. The 750 kg compulsator will generate 2.2 kV and the silicon-controlled rectifier (SCR) switch will commutate 386 kA pulses into the 1.6-m long, 0.60 caliber augmented solid armature railgun. The final design and predicted operating characteristics of the compulsator system are described. Overall system performance parameters are reported, including results from the optimization code used to aid in the design of the compulsator system. A system design overview is presented, with emphasis on new materials and state-of-the-art machine components to be used for the first time in a compulsator     

Complete rupture of the distal semimembranosus complex in a professional athlete

Complete ruptures of the distal hamstring tendons rarely cause functional disability significant enough to warrant surgical intervention. Isolated ruptures of the distal semimembranosus tendon complex have not been reported previously in the literature. A case of a complete rupture of the distal semimembranosus tendon complex occurring in a professional football player with resultant functional disability is reported here. The magnetic resonance imaging scan was essential for an accurate diagnosis. Subsequent surgical repair and postoperative rehabilitation is described. Strict attention to recreating the multiple attachments of the semimembranosus tendon complex is recommended.