PC and PE synthesis: mixed micellar analysis of the cholinephosphotransferase and ethanolaminephosphotransferase activities of human choline/ethanolamine phosphotransferase 1 (CEPT1)

The human choline/ethanolamine phosphotransferase 1 (CEPT1) gene codes for a dual-specificity enzyme that catalyzes the de novo synthesis of the two major phospholipids through the transfer of a phosphobase from CDP-choline or CDP-ethanolamine to DAG to form PC and PE. We used an expression system devoid of endogenous cholinephosphotransferase and ethanolaminephosphotransferase activities to assess the diradylglycerol specificity of CEPT1. A mixed micellar assay was used to ensure that the diradylglycerols delivered were not affecting the membrane environment in which CEPT1 resides. The CEPT1 enzyme displayed an apparent K _m of 36 μM for CDP-choline and 4.2 mol% for di-18∶1 DAG with a V _max of 14.3 nmol min⁻¹ mg⁻¹. When CDP-ethanolamine was used as substrate, the apparent K _m was 98 μM for CDP-ethanolamine and 4.3 mol% for di-18∶1 DAG with a V _max of 8.2 nmol min⁻¹ mg⁻¹. The preferred diradylglycerol substrates used by CEPT1 with CDP-choline as the phosphobase donor were di-18∶1 DAG, di-16∶1 DAG, and 16∶0/18∶1 DAG. A major difference between previous emulsion-based assay results and the mixed micelle results was a complete inability to use 16∶0(O)/2∶0 as a substrate for the de novo synthesis of platelet-activating factor when the mixed micelle assay was used. When CDP-ethanolamine was used as the phosphobase donor, 16∶0/18∶1 DAG, di-18∶1 DAG, and di-16∶1 DAG were the preferred substrates. The mixed micelle assay also allowed the lipid activation of CEPT to be measured, and both the cholinephosphotransferase and ethanolaminephosphotransferase activities displayed the unusual property of product activation at 5 mol%, implying that specific lipid activation binding sites exist on CEPT1. The protein kinase C inhibitor chelerythrine and the human DAG kinase inhibitor R59949 both inhibited CEPT1 activity with IC₅₀ values of 40 μM.     

Formation, structure, and rheological properties of ricinelaidic acid-vegetable oil organogels

Vegetable oil-based organogels were formed using ricinelaidic acid (12-hydroxy-9-trans-octadecenoic acid, REA). Gelation kinetics, gel structure, and stability were studied. Gelation occurred with as little as 0.5% (w/w) REA, depending on temperature and oil composition. Phase diagrams were constructed using canola, sesame, and DAG oils. Lower gelation tendencies were correlated with the presence of potential hydrogen-bonding moieties in the oils. REA concentration had a significant influence on gelation kinetics and gel rheology. At 5°C, the 0.5% canola oil gel behaved like a weak, viscoelastic network composed of entangled strands. Between 1.0 and 5.0% REA, solid-like, viscoelastic gels were formed. The 24-hour G′ _LVR (storage modulus in the linear viscoelastic region) was highly dependent on concentration and less so on temperature. Values for gelation time indicated a change in behavior below 2% REA and above 20°C. Polarized light microscopy revealed that the gels were formed through the interactions of long, thin, and birefringent fibers. Structural analysis using X-ray diffractometry (XRD) indicated the presence of repeating REA dimers and increasing order with concentration and gel storage time. Increases in gel opacity, birefringence, XRD scattering, and fiber clustering were observed during storage.     

Tripalmitin–Sodium Dodecyl Sulfate Emulsion Droplet Liquid vs. Solid State Impacts in vitro Digestive Lipolysis

Questions remain as to the impact of lipid structure, including crystallinity, on digestibility and metabolic response. This study was undertaken to determine the impact of triacylglycerol crystallinity on digestibility using undercooled (liquid emulsion, LE) and crystalline (solid emulsion, SE) particles exposed to an in vitro model simulating upper gastrointestinal tract (GIT) digestive conditions. By hot microfluidization, 10 wt% tripalmitin oil‐in‐water emulsions (D_3,2 ~ 0.115 nm) with 0.9 wt% sodium dodecyl sulfate (SDS) were prepared. SE demonstrated complex melting behavior, was predominantly in the beta polymorph, and consisted of a heterogeneous mixture of anisometric particles. In vitro duodenal lipolysis was more extensive for the spherically shaped LE droplets vs. SE (P < 0.05), despite the fact that exposure to simulated gastric conditions (at pH 2, but not at pH 7) induced partial crystallization. Therefore, lipid droplet physical state impacted and was impacted by exposure to gastrointestinal conditions, with differences observed in fatty acid digestive release and implications for lipid absorption.     

Spatial and seasonal variation in heavy metals in the sediments and biota of two adjacent estuaries, the Orwell and the Stour, in eastern England

A study was made of the concentrations of the elements As, Cd, Cu, Hg, Mn, Ni, Pb and Zn in the sediments and biota of two adjacent estuaries, the Orwell and Stour, in eastern England. The Orwell Estuary, with its urbanized head, was more contaminated with heavy metals than the Stour Estuary. Generally, in both estuaries, concentrations of metals were highest towards the head and the mouth. Saltmarsh sediments accumulated higher concentrations of most metals than mudflat sediments. Metal concentrations in the biota showed marked interspecific differences; Mytilus edulis had higher concentrations of Cd, Littorina littorea higher concentrations of Cu and Mn and Arenicola marina higher concentrations of Hg. Invertebrates from the Orwell had higher metal concentrations than those from the Stour. Algae had generally lower levels of metals than invertebrates. Metal concentrations were greatest and more variable in the top 10 cm of sediment. Metals were at greatest concentrations in winter and lowest in summer in sediments, algae and invertebrates. Mercury concentration increased with size in the three invertebrate species studied, but Cd and Zn generally were at higher concentrations in younger animals. Comparisons of sediments with average shale values indicated anthropogenic enrichment with several metals but it was considered that only Pb, at some sites, and possibly Hg posed potential threats to the ecology of the estuaries.     

Optimizing the Implementation of High-Temperature Fixed Points through the Use of Thermal Modeling

High-temperature fixed points (HTFP) have the potential to make a step-change improvement in high-temperature metrology, significantly reducing the uncertainty of scale realization of the current ITS-90 and improving dissemination of high-temperature scales to industry. However, in a practical implementation, the performance of HTFP could be limited, by, for example, injudicious use of insulation in the vicinity of the fixed point, furnace gradients, or incomplete filling. This article investigates some of these aspects for a selection of HTFP. Steady-state modeling of the influence of insulation on the radiance temperature was performed for Co–C (1,324°C), Pd–C (1,492°C), Pt–C (1,738°C), Ru–C (1,953°C), and Re–C (2,474°C) fixed points. This included studying mitigation scenarios through the insertion of different types and designs of insulation. The optimum design was identified to minimize the temperature drop in a particular furnace. It was found that, for the furnace and fixed-point combination modeled, the actual effect of the insulation was almost insignificant. Transient modeling was performed for a Re–C fixed point, to track the evolution of the radiance temperature through the melting transition. The starting point of the model was the beginning of the melt. The evolution of radiance temperature with time in “perfectly” filled cells was modeled with a range of linear temperature gradients across the eutectic cell. The gradient had a significant effect on the duration of the transition and on the structure of the melt itself. Despite the model’s simplicity, it qualitatively demonstrated that the melt transition temperature, as identified by the point of inflection, could be significantly affected by the presence of furnace gradients.     

Development of a rapid prototyping design advice system

This paper describes the initial development of a computer based Rapid Prototyping Design Advice System. The system is intended to assist the designer or project manager, particularly those in small and medium sized companies, in planning the prototyping stage of product development. It provides the user with an efficient and user friendly development aid which uses information obtained from the user and Computer Aided Design data to recommend suitable rapid prototyping solutions.     

Trends in health and ability to work among the older working-age population

OBJECTIVES: Because of recent changes in Social Security regulations that will soon begin to raise the age of eligibility for full retirement benefits, it is important to determine whether health and ability to work at older ages have improved in recent years. METHODS: Individual-level data from the National Health Interview Survey from 1982 through 1993 are used in this analysis. Trends in self-reported ability to work, presence of disease, and causes of actual work limitation are examined. RESULTS: Men and women in their 60s, that is those in the older working ages and younger retirement ages, report significant improvement in their ability to work. The change in work ability is large enough so the percentage unable to work at age 67 in 1993 is lower than the percentage unable to work at age 65 in 1982. This improvement appears to have been similar for racial and ethnic groups and across educational subgroups of the population, although African Americans and those with lower educational attainment are less healthy to begin with. The improvement in health is due to the changing educational composition of the population, which is linked to better life-long health, different occupational circumstances, and better health behaviors. In addition, the improvement in work ability is explained by decline in the prevalence of cerebro/cardiovascular diseases and arthritis. DISCUSSION: The level of observed improvement in work ability means that the legislated rise in age of full eligibility for Social Security benefits should be more than compensated for by the improved ability to work.