Rapid Quantification of Low-Viscosity Acetyl-Triacylglycerols Using Electrospray Ionization Mass Spectrometry

Acetyl‐triacylglycerols (acetyl‐TAG) possess an sn‐3 acetate group, which confers useful chemical and physical properties to these unusual triacylglycerols (TAG). Current methods for quantification of acetyl‐TAG are time consuming and do not provide any information on the molecular species profile. Electrospray ionization mass spectrometry (ESI–MS)‐based methods can overcome these drawbacks. However, the ESI–MS signal intensity for TAG depends on the aliphatic chain length and unsaturation index of the molecule. Therefore response factors for different molecular species need to be determined before any quantification. The effects of the chain length and the number of double‐bonds of the sn‐1/2 acyl groups on the signal intensity for the neutral loss of short chain length sn‐3 groups were quantified using a series of synthesized sn‐3 specific structured TAG. The signal intensity for the neutral loss of the sn‐3 acyl group was found to negatively correlated with the aliphatic chain length and unsaturation index of the sn‐1/2 acyl groups. The signal intensity of the neutral loss of the sn‐3 acyl group was also negatively correlated with the size of that chain. Further, the position of the group undergoing neutral loss was also important, with the signal from an sn‐2 acyl group much lower than that from one located at sn‐3. Response factors obtained from these analyses were used to develop a method for the absolute quantification of acetyl‐TAG. The increased sensitivity of this ESI–MS‐based approach allowed successful quantification of acetyl‐TAG in various biological settings, including the products of in vitro enzyme activity assays.     

Heterogenized copper catalysts for the amination of arylhalide: Synthesis,characterization and catalytic applications

This research investigated the synthesis, characterization and catalytic activity of heterogenized copper complexes for the selective amination of aryl halides. Amination reactions using homogeneous copper catalyst systems are not generally selective and it is also difficult to recover the catalyst from the homogeneous system for recycling. To overcome these difficulties, our efforts were directed towards the development of cheaper heterogeneous catalyst systems, which can be easily recovered and recycled. Heterogenized analogs of the homogenous Cu complexes were prepared using two methods: (a) encapsulation of copper complexes in zeolite-Y and (b) tethering of copper complexes on various supports like zeolite-Y, silica, charcoal, or clay. The encapsulated and tethered copper catalysts on zeolite-Y were characterized using EPR, Diffused reflectance UV–vis, XRD, IAS, ICPES, SEM, and TEM. The results confirmed that the copper complexes were truly heterogenized in zeolite-Y support. Encapsulated and tethered copper complexes were tested for amination of aryl iodide. The effects of various reaction parameters were examined using tethered Cu(Phen)(PPh₃)Br-PTA-Y catalyst to determine optimum conditions. A recycle study of encapsulated and tethered Cu catalyst was carried out to test their robustness for possible commercial exploitation.     

Evaluation of the impact of lipid fouling during the chromatographic purification of virus‐like particles from Saccharomyces cerevisiae

BACKGROUND: Production of recombinant virus‐like particles (VLPs) in yeast expression systems for use as vaccines requires cell disruption and detergent‐mediated steps to liberate the product. Typically, these release high levels of cellular components such as lipids that foul chromatography columns. This study compares the impact of applying lipid‐rich and lipid‐depleted feedstocks to hydrophobic interaction chromatography columns to quantify the loss of performance caused by the presence of host lipids over a total of 40 operational cycles. RESULTS: VLP binding capacity in the lipid‐rich feed was significantly lower than for the lipid‐depleted feed, with greater than 24% of the lipids remaining in the column after each cycle. Triacylglycerol was found to be the major contaminant. The effectiveness of subsequent caustic clean‐in‐place was limited, resulting in column hydrophobicity increasing over repeated loading cycles. This improved the effective VLP binding capacity and affinity, but also made product elution more difficult, and recovery decreased by more than 70% over the 40 cycles. CONCLUSION: Host cell lipids cause major fouling problems during VLP purification. Instead of screening for better CIP conditions, priority should be given to identifying suitable upstream lipid removal strategies in order to maintain column performance and so yield more economically viable processes. Copyright © 2009 Society of Chemical Industry     

The Yeast ATF1 Acetyltransferase Efficiently Acetylates Insect Pheromone Alcohols: Implications for the Biological Production of Moth Pheromones

Many moth pheromones are composed of mixtures of acetates of long‐chain (≥10 carbon) fatty alcohols. Moth pheromone precursors such as fatty acids and fatty alcohols can be produced in yeast by the heterologous expression of genes involved in insect pheromone production. Acetyltransferases that subsequently catalyze the formation of acetates by transfer of the acetate unit from acetyl‐CoA to a fatty alcohol have been postulated in pheromone biosynthesis. However, so far no fatty alcohol acetyltransferases responsible for the production of straight chain alkyl acetate pheromone components in insects have been identified. In search for a non‐insect acetyltransferase alternative, we expressed a plant‐derived diacylglycerol acetyltransferase (EaDAcT) (EC 2.3.1.20) cloned from the seed of the burning bush (Euonymus alatus) in a yeast system. EaDAcT transformed various fatty alcohol insect pheromone precursors into acetates but we also found high background acetylation activities. Only one enzyme in yeast was shown to be responsible for the majority of that background activity, the acetyltransferase ATF1 (EC 2.3.1.84). We further investigated the usefulness of ATF1 for the conversion of moth pheromone alcohols into acetates in comparison with EaDAcT. Overexpression of ATF1 revealed that it was capable of acetylating these fatty alcohols with chain lengths from 10 to 18 carbons with up to 27‐ and 10‐fold higher in vivo and in vitro efficiency, respectively, compared to EaDAcT. The ATF1 enzyme thus has the potential to serve as the missing enzyme in the reconstruction of the biosynthetic pathway of insect acetate pheromones from precursor fatty acids in yeast.     

Games to induce specified equilibria

Media access protocols in wireless networks require each contending node to wait for a backoff time, chosen randomly from a fixed range, before attempting to transmit on a shared channel. However, nodes acting in their own selfish interest may not follow the protocol. In this paper, a static version of the problem is modeled as a strategic game played by non-cooperating, rational players (the nodes). The objective is to design a game which exhibits a unique, a priori mixed-strategy Nash equilibrium. In the context of the media access problem, the equilibrium of the game would correspond to nodes choosing backoff times randomly from a given range of values, according to the given distribution. We consider natural variations of the problems concerning the number of actions available to the players and show that it is possible to design such a game when there are at least two players that each have the largest number of possible actions among all players. In contrast, we show that if there are exactly two players with different number of actions available to them, then it becomes impossible to design a strategic game with a unique such Nash equilibrium.     

Glass‐polymer melt hybrids. I: Viscoelastic properties of novel affordable organic‐inorganic polymer hybrids

A novel class of organic‐inorganic polymer hybrids was developed by melt‐blending up to 50 (v/v) % [about 83 (w/w) %] tin‐based polyphosphate glass (Pglass) and low‐density polyethylene (LDPE) in conventional plastics processing equipment. The liquid‐ and solid‐state rheology of the polymer hybrids was studied under oscillatory shear flow and deformation to understand the behavior of these materials and to accelerate efforts to melt process the Pglass with organic polymers. All the materials were found to be linearly viscoelastic in the range of temperature and frequencies examined and their viscoelastic functions increased with increasing Pglass concentration. The Pglass significantly enhanced the shear‐thinning characteristics of the Pglass‐LDPE hybrid, indicating the presence of nonlinear chemical and physical interactions between the hybrid components. Morphological examination of the materials by scanning electron microscopy revealed interesting evolution of microstructure of the Pglass phase from droplets (or round beads) to elongated and interpenetrating network structures as the glass concentration was increased in the Pglass‐LDPE hybrids. Melt viscosities of the materials were well described by a simple power‐law equation and a Maxwellian (Hookean) model with three relaxation times. Time‐temperature superpositioning (TTS) of the complex viscosity versus frequency data was excellent at 170°C < T < 220°C and the temperature dependencies of the shift factors conformed excellently well to predictions from an Arrhenius‐type relation, enabling calculation of the flow‐activation energies (25–285 kj/mol) for the materials. The beneficial function of the Pglass in the hybrid system was significantly enhanced by pre‐treating the glass with coupling agents prior to incorporating them into the Pglass‐LDPE hybrids.     

Investigation and simulation of a cross-flow air classifier

Extensive experimental tests and a computational study of the performance in a cross-flow air classifier have been carried out. A computational fluid dynamics (CFD) package—Fluent—is used to first understand and explain why the cuts or the sharpness of cut of this classifier are not as sharp as they ought to be, and then to optimize the geometry and operational conditions.

Flow fields of the classifier under various set-up conditions and geometry were measured by using laser Doppler anemometry (LDA). Using sieve analyses and the HELOS-laser method, the patterns of behaviour of separation parameters such as cut size and sharpness of cut have been investigated at different boundary conditions.

Using the Fluent package, a two-dimensional computational fluid dynamics model has been developed. The model is based on the Euler–Lagrangian approach. Different turbulence models have been tested. Both Fluent 4.5, with a structured grid, and Fluent 5.1, with structured and unstructured grids, have been used.

Discussions and analyses of the experimental, as well as the computational results, are presented. The simulation with a structured grid shows good agreement with experimental data, except for the sharpness of cut. The reasons of poor performance of the classifier have been found. The geometry is optimized and other conditions were also improved. The performance of the classifier is improved. The experimental observations together with the computed results should increase the depth of understanding of the underlying mechanisms.     

Regioselectivity in Hydroformylation of Aryl Olefins Using Novel Rhodium Polyether Phosphinite Catalysts

Polyethylene glycol based diphosphinite ligands and their Wilkinson type Rh-polymeric carbonyl complexes were prepared for homogeneous hydroformylation of styrenes. The effect of reaction variables such as temperature, pressure, P/Rh ratio, Substrate/Rh ratio, phosphine and phosphinite ligands on the activity and selectivity performance were discussed in detail. The catalyst showed a high rate of reaction, TON and tunable regioselectivity depending upon reaction conditions as compared to conventional Rh-phosphine based catalysts.     

Orientalism in Euro-American and Indian psychology: Historical representations of "natives" in colonial and postcolonial contexts.

The author examines the historical role of Euro-American psychology in constructing Orientalist representations of the natives who were colonized by the European colonial powers. In particular, the author demonstrates how the power to represent the non-Western "Other" has always resided, and still continues to reside, primarily with psychologists working in Europe and America. It is argued that the theoretical frameworks that are used to represent non-Westerners in contemporary times continue to emerge from Euro-American psychology. Finally, the author discusses how non-Western psychologists internalized these Orientalist images and how such a move has led to a virtual abandonment of pursuing "native" forms of indigenous psychologies in Third World psychology departments. (PsycINFO Database Record (c) 2010 APA, all rights reserved)