The Fifth WS/DGAT Enzyme of the Bacterium Marinobacter aquaeolei VT8

Wax esters (WE) belong to the class of neutral lipids. They are formed by an esterification of a fatty alcohol and an activated fatty acid. Dependent on the chain length and desaturation degree of the fatty acid and the fatty alcohol moiety, WE can have diverse physicochemical properties. WE derived from monounsaturated long-chain acyl moieties are of industrial interest due to their very good lubrication properties. Whereas WE were obtained in the past from spermaceti organs of the sperm whale, industrial WE are nowadays mostly produced chemically from fossil fuels. In order to produce WE more sustainably, attempts to produce industrial WE in transgenic plants are steadily increasing. To achieve this, different combinations of WE producing enzymes are expressed in developing Arabidopsis thaliana or Camelina sativa seeds. Here we report the identification and characterization of a fifth wax synthase from the organism Marinobacter aquaeolei VT8, MaWSD5. It belongs to the class of bifunctional wax synthase/acyl-CoA:diacylglycerol O-acyltransferases (WSD). The protein was purified to homogeneity. In vivo and in vitro substrate analyses revealed that MaWSD5 is able to synthesize WE but no triacylglycerols. The protein produces WE from saturated and monounsaturated mid- and long-chain substrates. Arabidopsis thaliana seeds expressing a fatty acid reductase from Marinobacter aquaeolei VT8 and MaWSD5 produce WE. Main WE synthesized are 20:1/18:1 and 20:1/20:1. This makes MaWSD5 a suitable candidate for industrial WE production in planta.     

Influence of accelerated ageing on methacrylate‐based denture resins heterogeneity as viewed by ESR‐spin‐probe method

The electron‐spin‐resonance (ESR) spin‐probe method, was used to study the heterogeneity of denture resins based on poly(methyl methacrylate). Results for three resins processed by microwave energy, conventional curing and cold curing (depending on the curing procedure and exposed to ageing in various environmental conditions) were compared. All three cured resins were stored over the same time (1200 h) in distilled water at ambient temperature and in artificial saliva at 348 K. The temperature‐dependent ESR spectra of a spin probe dispersed in the denture resins are analyzed in terms of line‐shapes and line‐widths. The appearance of two spectral components was taken as an indication of resin heterogeneity. The results reveal that the cold‐cured resin has a lower local density in comparison with microwave and conventionally cured resin. The amount of residual monomer also contributes to the local motion of polymer segments. The change of denture resins exposed to ageing is influenced both by the structure of the original resin and the ageing conditions. Restricted motion of a spin probe incorporated into the acrylic resins exposed to accelerated ageing suggests additional crosslinking of polymer chains. The differences are observed for all the investigated resins, but the highest change is observed with the cold‐cured resin. The ESR results are accompanied by T_g and T_m measurements. Copyright © 2005 Society of Chemical Industry     

The problem of the selection of an a posteriori error indicator based on smoothening techniques

This paper addresses the problem of assessing the quality of an a posteriori error estimate of a finite element solution. An error estimate based on local L2-projections is analysed in the case of translation-invariant meshes. It is shown that for general meshes this technique does not lead to an asymptotically exact estimator. The problem is analysed in detail in the one-dimensional setting. It is shown that an asymptotically exact estimator is not the optimal one when the solution is not sufficiently smooth. An optimal estimator for adaptively constructed meshes is given. Finally, a general mathematical framework for the quality assessment of estimators is introduced.     

Recrystallization/precipitation behaviour in microalloyed steels

Microalloyed high-strength low-alloy (HSLA) steels contain additions of Nb, V, Ti, or in combination, in amounts of 0.01 to 0.1 weight percent to improve mechanical properties, which are strongly dependent on the thermomechanical interaction taking place in the course of rolling mill processes. The recrystallizatian of hat-twisted austenite has been investigated in a cylindrical specimen (f 6×50 mm) machined from hat rolled plates of 0,052 wt % Niobium microalloyed steel. Continuous and interrupted torsion test were carried out in the temperature range 1123 K to 1173 K after a solution treatment of 1.5 minutes at 1423 K and torque-twist data were analysed. The various methods were discussed for obtaining results from torsion tests. The effect of precipitation kinetics was appreciated by way of connection t_p/t_p(red), where t_p is the experimental measured time for the peak stress and t_p(red) is the newly defined reduced time. The softening ratio X and time t_0.05R for start of static recrystallization were established.

The correlation between precipitation and recrystallization is presented as a graphs for chosen requirements (temperature of austenitization, carbon and niobium content and strain rate). If temperature goes below 850°C, the restoration processes are hardly suppressed, both are limited by diffusion and Nb(CN) precipitation, which are extended dynamically in the range of strains rates 10⁻² to 1 s⁻¹.

In the present paper, an attempt is made to derive the PRTT diagram and to define all mathematical equations for describing recrystallization times t_0.05R, t_0.5R, t_0.95R and t_0.05P for the start of precipitation. In real metal forming processes such as the hot rolling of plates or strips the knowledge of these parameters and results is extremely important for the the correct microstructure and sheet quality to be obtained.     

Magnetic force microscopy studies on the magnetic ordering in organic materials induced by high-energy proton irradiation

In this study, ferromagnetic microstructures in highly oriented pyrolytic graphite and superparamagnetic spots in polyimide foils were created by 2.25 MeV proton microbeam irradiation and characterized using atomic and magnetic force microscopy. For this purpose, graphite samples were irradiated with cross-like patterns of 15 μm × 15 μm size using ion fluences in the range of (0.003–2.5) × 10¹⁸ cm⁻². The irradiated crosses showed strong magnetic signals and a complex domain structure in the magnetic images depending on the geometrical dimensions of the crosses. Furthermore, polyimide foils were irradiated with microspots and fluences in the range of (0.016–3.1) × 10¹⁹ cm⁻². Magnetic force microscopy shows very strong phase shifts in these irradiated areas.     

Effects of selenium and vitamin E on incidence of retained placenta

In a series of experiments over 3 yr, relatively low doses of selenium and vitamin E were injected intramuscularly 3 wk prepartum to pregnant Israeli-Holstein dairy cows in a high-producing herd (9000 kg milk/yr) with a history of retained placenta in 17% of the primiparous and 28% of the multiparous animals, as recorded for the 12 mo preceding the study. Selenium ranged from .035 to .109 ppm in the prepartum diet and from .160 to .200 ppm in the postpartum diet. Doses of selenium ranging from 2.3 to 23.0 mg reduced the incidence of retained placenta in 186 primiparous and in 428 multiparous to 7 and 15%, which was half of that of controls. Low doses of selenium (2.3 to 4.6 mg) tended to be more effective than higher ones. Selenium alone was at least as effective as a combination of selenium and vitamin E. Dosage, timing, mechanism, and interactions of selenium with other factors in reducing the incidence of retained placenta need clarification.     

Design and development of a multichannel potentiometer for monitoring an electrode array and its application in flow analysis

A versatile potentiometer that works with electrode arrays in flow injection and/or monosegmented flow systems is described. The potentiometer is controlled by a microcomputer that allows individual, sequential multiplexed or random accesses to eight electrodes while employing only one reference electrode. The instrument was demonstrated by monitoring an array of seven flow-through ion-selective electrodes for Ag(+) and for three electrodes for Cl(-), Ca(2+) and K(+). The figures of merit of the individual and multiplexed (summed) readings of the electrode array were compared. The absolute standard deviation of the measurements made by summing the potential of two or more electrodes was maintained constant, thus improving the precision of the measurements. This result shows that an attempt to combine the signals of the electrodes to produce a more intense signal in the Hadamard strategy is feasible and accompanied by a proportional improvement in the precision of individual measurements. The preliminary tests suggest that the system can allow for 270 determinations per hour, with a linear range from 1.0 x 10(-2) to 1.0 x 10(-4) mol l(-1) for the three di inverted exclamation markerent analytes. Detection limits were estimated as 3.1 x 10(-5), 3.0 x 10(-6) and 1.0 x 10(-5) mol l(-1) for Cl(-), Ca(2+) and K(+), respectively.     

Serotonin-4 receptor antagonists reverse cocaine-induced cardiac arrhythmia

CONTEXT: Adolescent smoking prevalence is tracked annually and has increased since 1991. In contrast, little is known about trends in smoking among college students, a group that has previously been more resistant to tobacco use than other young adults. OBJECTIVE: To examine changes in cigarette smoking among college students between 1993 and 1997 and among different types of students and colleges. DESIGN: Self-administered survey (Harvard School of Public Health College Alcohol Study). SETTING: One hundred sixteen nationally representative 4-year colleges. SUBJECTS: A total of 15103 randomly selected students in 1993 (70% response rate) and 14251 students in 1997 (60% response rate). MAIN OUTCOME MEASURES: Self-reports of cigarette smoking in the past 30 days and in the past year, age at smoking first cigarette, and number of attempts to quit. RESULTS: Over 4 years, the prevalence of current (30-day) cigarette smoking rose by 27.8%, from 22.3% to 28.5% (P<.001). The increase was observed in 99 of 116 colleges and was statistically significant (P<.05) in 27 (23%) of them. Current smoking increased across all student subgroups (defined by sex, race/ethnicity, and year in school) and in all types of colleges. Smoking is rising faster in public schools (from 22.0% to 29.3%) than in private schools (from 22.9% to 26.8%). Eleven percent of college smokers had their first cigarette and 28% began to smoke regularly at or after age 19 years, by which time most were already in college. Half of current smokers tried to quit in the previous year; 18% had made 5 or more attempts to quit. CONCLUSIONS: Cigarette use is increasing on campuses nationwide in all subgroups and types of colleges. Substantial numbers of college students are both starting to smoke regularly and trying to stop. National efforts to reduce smoking should be extended to college students.     

Low strain rate plasticity of continuously cast carbon steels

By a special plastometric method, the straightening operation of a vertically cast strand of two carbon steels was simulated. The temperature dependence of the low strain rate plasticity of both the steels examined is analogous although caused by very different reasons. As to the plain carbon steel, the amount of ferrite and its location is of prime importance, whereas in the case of eutectoid steel various modes of fracture and the influence of temperature on the relative plasticity of type II MnS inclusions are influencial. To avoid transverse cracking, it seems to be suitable to keep the strand at a rather high temperature (above 900 °C). Temperature cycling seems to be fundamental as to the structural changes taking place in the strand (grain size refining, ferrite fraction if any), whereas analogous influences of straining are more or less inexpressive. Of course, the results obtained are of quality only since the practical and laboratory conditions differ significantly. The knowledge obtained could be applied in regulating the intensity of strand cooling with the aim of lowering the waste portion and improving the surface quality of worked products.     

External surface and lamellarity of lipid vesicles: a practice-oriented set of assay methods

Three methods are presented for the determination of external surface of large lipid vesicles of different lamellarity with 2% absolute accuracy. These methods (referred to as EPR, NBD and TNBS assays) use different marker lipids which provide signals (electron paramagnetic resonance, fluorescence of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) residues, and UV absorption increase of 2,4,6-trinitrobenzenesulfonic acid after reaction with aminolipids, respectively). The signals change upon addition of different membrane-impermeant reagents due to reaction with marker lipids at the external vesicle surface. They were applied to the same vesicle samples, including unilamellar and multilamellar vesicles, both at two different lipid compositions. External surface data matched for the three assays within 2%, but only after appropriate redesign or adaptation of so far published procedures. Main improvements related to slow influx of reagents (TNBS and NBD assays) or to redistribution of marker lipids (EPR assay), obscuring determination of outer vesicle surface from fast reaction between reagent and readily accessible marker lipids. Furthermore, suitable strategies were found to obtain accurate 100% values (reaction of all marker lipids present), required to relate external vesicle surface to total surface. This included corrections for light scattering (NBD assay) and for turbidity (TNBS assay). These three methods appear to close a gap in the methodology to determine external surface of vesicles for typical practical needs. In particular, the reliability range of the NBD assay could be extended to marker lipid densities as low as 1 marker lipid per 3000 lipids.