Two structural domains of initiation factor eIF-4B are involved in binding to RNA

Translation initiation factor eIF-4B promotes the binding of mRNA to 40 S preinitiation complexes and together with eIF-4A possesses RNA helicase activity. To elucidate structural features involved in its function, a series of internal and C-terminal deletions, as well as point mutations, were constructed in the eIF-4B cDNA. The mutated cDNAs were expressed in transiently transfected COS-1 cells, and mutant forms of the factor were overproduced up to about 25-fold over endogenous eIF-4B levels. Inhibition of dihydrofolate reductase (DHFR) synthesis by high levels of eIF-4B variants was determined in vivo, and the binding of the eIF-4B forms to biotinylated RNA was measured in vitro. The results indicate that the N-terminal region containing the RNA binding motif with its RNP1 and RNP2 consensus elements is sufficient for inhibition of DHFR synthesis. Deletion of the RNP1 sequence abrogates RNA binding, but amino acid substitutions at conserved residues do not always inhibit RNA binding. Deletion of the DRYG domain near the middle of eIF-4B results in inhibition of RNA binding, but not of DHFR synthesis. Up to 164 residues of the C terminus are not required for RNA binding, but removal of 226 or more residues completely inhibits RNA binding, perhaps by the loss of two arginine-rich regions. The results suggest that both the RNA recognition motif and the arginine-rich region are required for stable RNA binding but that both are not necessary for in vivo inhibition of protein synthesis.     

Parallelisation of suboptimal spectrum search

The authors suggest that effective algorithms for spectrum search, such as those used for detecting spread spectrum signals, may be derived by selecting suboptimal algorithms and then recovering some of the lost efficacy through parallelisation methods. This thesis is motivated by considering a simple yet meaningful example of a spectrum search technique that exhibits what at first may seem to be counterintuitive behaviour.<>     

Error rate for optimal follower tone-jamming

The performance of a countermeasure technique in the presence of the optimal follower multitone jammer is evaluated for frequency-hopped spread spectrum (FHSS) communications. It is shown that, with a certain probability, the optimal jammer will have dual tones in a frequency channel     

Comparison of three rewarming methods in a postanesthesia care unit

Postoperative hypothermia is problematic because patients in postanesthesia care units (PACUs) often feel very cold, and unrecognized or prolonged postoperative hypothermia can aggravate patients' underlying cardiovascular disorders. The researchers compared three methods of rewarming PACU patients who had undergone laparotomy procedures. Patients were assigned randomly to three groups. Each patient in group one received the standard PACU rewarming intervention (ie, two warmed thermal blankets and a hospital bedspread). Each patient in group two received the standard PACU rewarming intervention plus a reflective blanket. Each patient in group three received the standard PACU rewarming intervention plus a reflective blanket and a reflective head covering. Nurses measured patients' vital signs on admission to the PACU and every 15 minutes thereafter until patients' sublingual temperatures reached 36 degrees C (96.8 degrees F). No significant temperature differences occurred among patients in the three groups, but an inverse relationship existed between patients' PACU admission temperatures and the time they required to reach normothermia.     

Numerical study of steady turbulent flow through bifurcated nozzles in continuous casting

Bifurcated nozzles are used in continuous casting of molten steel, where they influence the quality of the cast steel slabs. The present study performs two-dimensional (2-D) and three-dimensional (3-D) simulations of steady turbulent(K- ε) flow in bifurcated nozzles, using a finite-element (FIDAP) model, which has been verified previously with water model experiments. The effects of nozzle design and casting process operating variables on the jet characteristics exiting the nozzle are investigated. The nozzle design parameters studied include the shape, angle, height, width, and thickness of the ports and the bottom geometry. The process operating practices include inlet velocity profile and angle as well as port curvature caused by erosion or inclusion buildup. Results show that the jet angle is controlled mainly by the port angle but is steeper with larger port area and thinner walls. The degree of swirl is increased by larger or rounder ports. The effective port area, where there is no recirculation, is increased by smaller or curved ports. Flow asymmetry is more severe with skewed or angled inlet conditions or unequal port sizes. Turbulence levels in the jet are higher with higher casting speed and smaller ports. Formerly Research Assistant, Department of Mechanical and Industrial Engineering. Formerly Research Assistant in the same department.     

Synthesis and triplex forming properties of an acyclic N7-glycosylated guanine nucleoside

A chiral acyclic nucleoside, one in which the ribose carbohydrate has been replaced with a glycerol-based linker, is prepared by glycosylating guanine at the N7-nitrogen. The stereochemically pure derivative is converted to a DMT-protected phosphoramidite for incorporation into DNA sequences. Sequence containing the acyclic N7-dG nucleoside are capable of forming DNA triplexes in which it is likely that the N1-H and N2-amino groups of the N7-dG are involved in recognition of the guanine base in G-C base pairs.     

Evaluation of chromic oxide,glycerol triether,and β-sitosterol as fecal flow markers in two species of nonhuman primates

Recovery of β-sitosterol, glycerol triether (1-hexadecyl-2,3,-didodecyl glycerol triether), and chromic oxide was studied in African green monkeys and stumptail macaques consuming diets containing 0.75 mg/Cal cholesterol and 38% of calories as safflower oil or butter. Following oral administration of these compounds, feces were collected daily for 9 days. For all animals, excretion of β-sitosterol and glycerol triether paralleled one another almost exactly. Except for two animlas, this was also true for chromic acid. Essentially 100% of the administered β-sitosterol and 90–95% of the glycerol triether were recovered; excretion of these markers virtually was complete by day 3. Ninety-two percent of the β-sitosterol was isolated in the nonsaponifiable lipid extract of the feces with less than 6% in the remaining aqueous phase. A maximum of 3.1% of the β-sitosterol and 1.8% of the glycerol triether were found in the blood. For stumptail macaques, the major excretory form of β-sitosterol was the 5β-derivative. African green monkeys were more variable; one animal excreted the bulk of the β-sitosterol unchanged while others excreted greater than 80% as the ring-saturated 5β-derivative. Animals consuming the safflower oil containing diet consistently excreted a greater percentage of the β-sitosterol unchanged, compared with those animals eating the butter containing diet. There was no evidence for steroid ring degradation in any of the animals used in this study.     

Functional Chromatography Reveals Three Natural Products that Target the Same Protein with Distinct Mechanisms of Action

Access to lead compounds with defined molecular targets continues to be a barrier to the translation of natural product resources. As a solution, we developed a system that uses discrete, recombinant proteins as the vehicles for natural product isolation. Here, we describe the use of this functional chromatographic method to identify natural products that bind to the AAA+ chaperone, p97, a promising cancer target. Application of this method to a panel of fungal and plant extracts identified rheoemodin, 1‐hydroxydehydroherbarin, and phomapyrrolidone A as distinct p97 modulators. Excitingly, each of these molecules displayed a unique mechanism of p97 modulation. This discovery provides strong support for the application of functional chromatography to the discovery of protein modulators that would likely escape traditional high‐throughput or phenotypic screening platforms.     

Preparation and properties of silane-endcapped polyimide adhesives

Silane-endcapped polyimide high temperature adhesive formulations were prepared by reacting anhydride-terminated poly(amic acid), obtained from benzophenonetetracarboxylic dianhydride and a diamine (3,3′-, 3,4′- or 4,4′ -diaminodiphenylmethane and 3,3′, 3,4′- or 4,4′ -diaminobenzophenone) with varying amounts of γ-aminopropyltriethoxysilane in dimethylacetamide. Resin properties were evaluated by torsional braid analysis and thermogravimetric analysis. Lap shear strengths of some of the adhesive bonds were determined at room temperature and at 177°C before and after ageing at 200°C for 2500 h and after boiling in water for 72 h.     

Structure-property relationships of isomeric addition polyimides containing nadimide end groups

Addition polyirnide oligomers with nadimide end groups (I) have been synthesized from 4,4′-oxydiphthalic anhydride and 3,3′,4,4′-benzophenone tetracarboxylic acid dianhydride with several isomeric diamines and, nadic anhydride. The low molecular weight amic acid? and corresponding imides were isolated and characterized. Solubility and melt-flow properties of the imide prepolymers were studied to determine the applicability of the resins as adhesives and composite matrices. Thermomechanical transitions of the polymers were obtained by torsional braid analysis. Properties were compared with a similar addition polymer, P13N.