Phosphorylation Stoichiometries of Human Eukaryotic Initiation Factors

Eukaryotic translation initiation factors are the principal molecular effectors regulating the process converting nucleic acid to functional protein. Commonly referred to as eIFs (eukaryotic initiation factors), this suite of proteins is comprised of at least 25 individual subunits that function in a coordinated, regulated, manner during mRNA translation. Multiple facets of eIF regulation have yet to be elucidated; however, many of the necessary protein factors are phosphorylated. Herein, we have isolated, identified and quantified phosphosites from eIF2, eIF3, and eIF4G generated from log phase grown HeLa cell lysates. Our investigation is the first study to globally quantify eIF phosphosites and illustrates differences in abundance of phosphorylation between the residues of each factor. Thus, identification of those phosphosites that exhibit either high or low levels of phosphorylation under log phase growing conditions may aid researchers to concentrate their investigative efforts to specific phosphosites that potentially harbor important regulatory mechanisms germane to mRNA translation.     

A concurrent two-layer restoration scheme for GMPLS WDM networks

Next generation backbone networks will likely consist of IP routers as well as optical cross connects (OXCs) and will deploy an optical control plane protocol. Generalized Multi Protocol Label Switching (GMPLS) has been proposed as the candidate of choice for the control plane. Optical fibers may carry large volumes of traffic and therefore adequate mechanisms must exist to enable the network to automatically recover from failures of fiber. In mission critical networks survivability becomes very important. We investigate the problem of autonomous recovery in such networks. The literature contains work in this area that investigates the problem of multilayer recovery. Such recovery had only been sequential in the sense that the published work recovers first in the optical domain, assuming the availability of redundant resources, and then proceeds to recover packet label switched paths. We report a recovery procedure for recovering packet label switch paths (packet LSPs) and lambda label switch paths (λLSP) concurrently. We have conducted an OPNET-based simulation study that compares the performance of the concurrent scheme with the previously published sequential two-layer recovery scheme. The study shows that the concurrent two-layer recovery scheme performs as much as forty-four percent faster than the sequential two-layer recovery scheme.     

Determination of nonlinear electromechanical energy conversion and the field-dependent elastic modulus of Pb(Mg(1/3)Nb(2/3 ))O(3)-based electrostrictors

Energy conversion efficiency is a critical parameter for all electromechanical materials. Although excellent techniques are available for linear materials, nonlinearities complicate the determination of conversion efficiency in electrostrictive Pb(Mg(1/3)Nb(2/3 ))O(3) (PMN). The field dependence of the elastic modulus presents additional problems. A technique combining pulse-echo ultrasound has been developed to approximate the change in the Young's modulus with applied field. A 30% decrease was observed at 1 MV/m (from ~100 to ~70 GPa). The boundary condition for these measurements was a constant electric field as opposed to constant dielectric displacement. In combination with the results from harmonic analysis, the modulus data may be used to determine the electromechanical conversion efficiency. This has been accomplished using an energy balance criterion for a PMN-based composition (with and without dc bias). The resulting longitudinal coupling values are >0.5 for practical electric fields (<1 MV/m peak).     

Response of serum cytokines in patients undergoing laparoscopic cholecystectomy

The clinical observation that a laparoscopic cholecystectomy is a minimally invasive operation has not been demonstrated on a biochemical basis. Interleukin-6, a known endogenous pyrogen and hepatocyte-stimulating protein, correlates with the significance of surgical trauma. Utilizing the IL-6 immunoassay, we studied this biochemical parameter of trauma to compare its response in laparoscopic vs open cholecystectomy. Sixteen patients who underwent only laparoscopic cholecystectomy showed peak IL-6 concentrations of 51 pg/ml (22-86) vs a peak IL-6 concentration of 124 pg/ml (56-225) for open cholecystectomy. Six additional patients who underwent an ERCP followed by laparoscopic cholecystectomy showed a dramatic rise in peak IL-6 concentration to 315 pg/ml (15-634). These results biochemically confirm the true minimal invasiveness of laparoscopic cholecystectomy. The findings in the ERCP-followed-by-laparoscopic-cholecystectomy group support the theory that two invasive procedures in close proximity may prime the cytokine system in its response to surgical trauma.     

A probability matrix for the identification of campylobacters, helicobacters and allied taxa

A probabilistic identification matrix for campylobacteria, comprising 76 phenotypic characters and 37 taxa, is described. The accuracy and integrity of the matrix was evaluated using established computer-assisted methods. Certain taxa (for example, Campylobacter concisus and Camp. gracilis) demonstrated significant phenotypic diversity; previous data corroborated these findings. Differentiation between a few pairs of taxa proved difficult, although discriminatory characteristics were noted in each of these cases. The results indicate that most campylobacteria can be identified accurately and objectively with phenotypic tests when probabilistic methods of data assessment are employed.     

Primary pneumococcal peritonitis complicated by exudative pleural effusion in an adolescent girl

A healthy, young adolescent girl developed primary pneumococcal peritonitis, an infection rarely reported in this age group in North America. Her course was further complicated by exudative pleural effusion and pneumonia despite receiving 10 days of clindamycin therapy. Laparascopy proved useful in making the initial diagnosis, but may have contributed to the pathogenesis of the pulmonary process. Case presentation, management, and etiology are discussed.     

Mapping of a yeast G protein betagamma signaling interaction

The mating pathway of Saccharomyces cerevisiae is widely used as a model system for G protein-coupled receptor-mediated signal transduction. Following receptor activation by the binding of mating pheromones, G protein betagamma subunits transmit the signal to a MAP kinase cascade, which involves interaction of Gbeta (Ste4p) with the MAP kinase scaffold protein Ste5p. Here, we identify residues in Ste4p required for the interaction with Ste5p. These residues define a new signaling interface close to the Ste20p binding site within the Gbetagamma coiled-coil. Ste4p mutants defective in the Ste5p interaction interact efficiently with Gpa1p (Galpha) and Ste18p (Ggamma) but cannot function in signal transduction because cells expressing these mutants are sterile. Ste4 L65S is temperature-sensitive for its interaction with Ste5p, and also for signaling. We have identified a Ste5p mutant (L196A) that displays a synthetic interaction defect with Ste4 L65S, providing strong evidence that Ste4p and Ste5p interact directly in vivo through an interface that involves hydrophobic residues. The correlation between disruption of the Ste4p-Ste5p interaction and sterility confirms the importance of this interaction in signal transduction. Identification of the Gbetagamma coiled-coil in Ste5p binding may set a precedent for Gbetagamma-effector interactions in more complex organisms.     

Effects of preliminary muscular exercise on body temperature, water loss and physical performance

We investigated the effects of preliminary exercise (muscular warm-up) on body temperature, water loss and physical performance during consecutive sustained exercise. Thirty-one untrained men aged 21 to 30 years old (mean 25.12 +/- 2.92) were subjected to two physical trial tests at 75% Pma. One trial. (T - PE) was performed without preliminary exercise (PE) and the other (T + PE) was preceded by 15 minutes of preliminary exercise performed at 50% Pma. The trials involved pedaling an ergocycle until exhaustion, followed by a 30 minutes period of inactive recovery. The rate of increase of body temperature during the work consecutive to preliminary exercise (T + PE) was lower than that of the work without preliminary exercise (T - PE). The energy output and water loss during T + PE were significantly (P < 0.01) greater than during T - PE. However, the body temperatures at the end of the two tests were identical. The rate of decrease of body temperature, measured after 30 minutes of recovery, was higher for T + PE than T - PE. The duration of work was increased by PE for 25 (80.65%) subjects and decreased for 6 (19.35%). We conclude that preliminary exercise allows better adjustment of thermohydric regulation by moderating the rise in body temperature and increasing water loss during physical work. For most subjects, these adjustments allow improved endurance.