Prediction of coal ash fusion temperatures with the F*A*C*T thermodynamic computer package

This paper outlines research on the processes taking place within the coal mineral matter at high temperatures and development of the relationship between ash fusion temperatures (AFT) and phase equilibria of the coal ash slags. A new thermodynamic database for the Al-Ca-Fe-O-Si system developed by the author was used in conjunction with the thermodynamic computer package F*A*C*T for these purposes. In addition, high temperature experimental studies were undertaken that involved heat treatment and quenching of the ash cones followed by the analyses using different techniques. The study provided new information on the processes taking place during AFT test and demonstrated the validity of the AFTs predictions with F*A*C*T. Examples of practical applications of the AFT prediction method are given in the paper. The results of this study are important not only for the AFT predictions, but also in general for the application of phase equilibrium science to the characterisation of the coal mineral matter interactions at high temperature. (C) 2002 Elsevier Science Ltd. All rights reserved.

The microbiological quality of beef tripe using different processing techniques

The microbiological quality of routinely processed tripe and rumen pillars were compared with those derived after emptying the rumen (paunch) without using water (dry dumping) and after deliberately bursting the paunches before processing. Prior to packing the mean:log(10) aerobic plate counts (APC) for the routinely processed tripe and rumen pillars were 3.55+/-1.08 and 3.28+/-0.87/g respectively. The corresponding mean log(10) total coliform counts (TCC) were 1.27+/-1.28 and 2.08+/-0.87. The mean log(10) APC counts on tripe and rumen pillars after dry-dumping were 3.06+/-0.60 and 3.90+/-0.75/g, respectively. The corresponding mean log(10) TCC were 1.03+/-0.60/g and 2.75+/-1.14/g respectively. After deliberately bursting the paunches, before processing, the mean log(10) APC counts on tripe and rumen pillars were 3.55+/-0.83/g and 3.50+/-0.59/g and the mean log(10) TCC were 1.54+/-0.95/g and 2.66+/-0.82/g respectively. In all cases the prevalence of Salmonella and Campylobacter spp. was less than 3%. The results indicate that both tripe and rumen pillars can be produced after dry dumping without compromising the quality of tripe and rumen pillars. Similarly, incidentally burst paunches that become contaminated with ingesta on the serosal surface can be processed without compromising product quality. Crown Copyright (C) 2002 Published by Elsevier Science Ltd. All rights reserved.

Conformational changes in monolayers of a cationic amphiphilic porphyrin on saline subphases

Langmuir monolayer films of the tetracationic porphyrin tetrakis(octadecyl-4-pyridin ium)porphyrinatozinc(II) bromide on various salt containing subphases were analyzed using surface pressure-area isotherms and X-ray reflectivity. The use of these complementary techniques showed that the porphyrin molecules undergo changes in conformation upon compression. Two main phases were identified, one in which the porphyrin moiety is parallel to the subphase and one in which the porphyrin moiety is tilted out of the plane. The addition of different salts into the subphase brought about changes in film behaviour, which are explained in terms of a lyotropic series. Copyright (C) 2002 Society of Porphyrins,& Phthalocyanines.

Production of the baculovirus-expressed dengue virus glycoprotein NS1 can be improved dramatically with optimised regimes for fed-batch cultures and the addition of the insect moulting hormone, 20-Hydroxyecdysone

A perennial problem in recombinant protein expression is low yield of the product of interest. A strategy which has been shown to increase the production of baculovirus-expressed proteins is to utilise fed-batch cultures. One disadvantage of this approach is the time-consuming task of optimising the feeding strategy. Previously, a statistical optimisation routine was applied to develop a feeding strategy that increased the yield of beta-Galactosidase (beta-Gal) by 2.4-fold (Biotechnol. Bioeng, 59 (1998) 178). This involves the single addition of nutrient concentrates (amino acids, lipids. glucose and yeastolate ultrafiltrate) into Sf9 cell cultures grown in SF900II medium. In this study, it is demonstrated that this optimised fed-batch strategy developed for a high-yielding intracellular product beta-Gal could be applied successfully to a relatively low-yielding glycosylated and secreted product such as the dengue virus glycoprotein NS1. Optimised batch infections yielded 4 mug/ml of NS1 at a peak cell density of 4.2 x 10(6) cells/ml. In contrast. optimised fed-batch infections exhibited a 3-fold improvement in yield, with 12 mug ml of NS1 produced at a peak cell density of 11.3 x 10(6) cells/ml. No further improvements in yield were recorded when the feed volumes were doubled and the peak cell density was increased to 23 x 10(6) cells/ml, unless the cultures were stimulated by the addition of 4 mug/ml of 20-Hydroxyecdysone (an insect moulting hormone). In this case, the NS1 yield was increased to 20 mug/ml. which was nearly 5-fold higher than optimised batch cultures. (C) 2002 Elsevier Science B.V. All rights reserved.

Spontaneous generation and survival of blood dendritic cells in mononuclear cell culture without exogenous cytokines

Studies on purified blood dendritic cells (DCs) are hampered by poor viability in tissue culture. We, therefore, attempted to study some of the interactions/relationships between DCs and other blood cells by culturing unseparated peripheral blood mononuclear cell (PBMC) preparations in vitro. Flow cytometric techniques were used to undertake a phenotypic and functional analysis of DCs within the cultured PBMC population. We discovered that both the CD11c(+) and CD11c(-) CD123(hi) DC subsets maintained their viability throughout the 3-day culture period, without the addition of exogenous cytokines. This viability was accompanied by progressive up-regulation of the surface costimulatory (CD40, CD80, CD86) and activation (CMRF-44, CMRF-56, CD83) molecules. The survival and apparent production of DCs in PBMC culture (without exogenous cytokines) and that of sorted DCs (with cytokines) were evaluated and compared by using TruCOUNT analysis. Absolute DC counts increased (for CD123hi and CD11c+ subsets) after overnight culture of PBMCs. Single-cell lineage depletion experiments demonstrated the rapid and spontaneous emergence of new in vitro generated DCs from CD14(+)/CD16(+) PBMC radioresistant precursors, additional to the preexisting ex vivo DC population. Unlike monocyte-derived DCs, blood DCs increased dextran uptake with culture and activation. Finally, DCs obtained after culture of PBMCs for 3 days were as effective as freshly isolated DCs in stimulating an allogeneic mixed leukocyte reaction. (C) 2002 by The American Society of Hematology.

Multiple maps and activity-dependent representational plasticity in the anterior Wulst of the adult barn owl (Tyto alba)

In the present study we addressed the issue of somatosensory representation and plasticity in a nonmammalian species, the barn owl. Multiunit mapping techniques were used to examine the representation of the specialized receptor surface of the claw in the anterior Wulst. We found dual somatotopic mirror image representations of the skin surface of the contralateral claw. In addition, we examined both representations 2 weeks after denervation of the distal skin surface of a single digit. In both representations, the denervated digital representation became responsive to stimulation of the adjacent, mutually functional, digit. The mutability and multiple representations indicates that the Wulst provides the owl with sensory processing capabilities analogous to those in mammals.

Effect of Co addition on the lattice parameter, electrical conductivity and sintering of gadolinia-doped ceria

Co-sintering aid has been added to Ce1.9Gd0.1O1.95 (CGO) by treating a commercial powder with Co(NO3)(2) (COCGO), X-ray diffraction (XRD) measurements of lattice parameter indicated that the Co was located on the CGO particle surface after calcination at 650 degreesC. After heat treatment at temperatures above 650 degreesC, the room temperature lattice parameter of CGO was found to increase, indicating redistribution of the Gd. Compared to CGO, the lattice parameter of CGO + 2 cation% Co (2CoCGO) was lower for a given temperature (650-1100 degreesC), A.C. impedance revealed that the lattice conductivity of 2CoCGO was enhanced when densified at lower temperatures, Transmission electron microscopy (TEM) showed that, even after sintering for 4 h at 980 degreesC, most of the Co was located at grain boundaries. (C) 2002 Published by Elsevier Science B.V.

Analytical exclusion chromatography

This review summarizes the development of exclusion chromatography, also termed gel filtration, molecular-sieve chromatography and gel permeation chromatography, for the quantitative characterization of solutes and solute interactions. As well as affording a means of determining molecular mass and molecular mass distribution, the technique offers a convenient way of characterizing solute selfassociation and solute-ligand interactions in terms of reaction stoichiometry and equilibrium constant. The availability of molecular-sieve media with different selective porosities ensures that very little restriction is imposed on the size of solute amenable to study. Furthermore, access to a diverse array of assay procedures for monitoring the column eluate endows analytical exclusion chromatography with far greater flexibility than other techniques from the viewpoint of solute concentration range that can be examined. In addition to its widely recognized prowess as a means of solute separation and purification, exclusion chromatography thus also possesses considerable potential for investigating the functional roles of the purified solutes. (C) 2003 Elsevier Science B.V. All rights reserved.

Purification and characterisation of functional early pregnancy factor expressed in Sf9 insect cells and in Escherichia coli

Early pregnancy factor (EPF) is a secreted protein with growth regulatory and immunomodulatory properties. It is an extracellular form of the mitochondrial matrix protein chaperonin 10 (Cpn10), a molecular chaperone. An understanding of the mechanism of action of EPF and an exploration of therapeutic potential has been limited by availability of purified material. The present study was undertaken to develop a simple high-yielding procedure for preparation of material for structure/function studies, which could be scaled up for therapeutic application. Human EPF was expressed in Sf9 insect cells by baculovirus infection and in Escherichia coli using a heat inducible vector. A modified molecule with an additional N-terminal alanine was also expressed in E coli. The soluble protein was purified from cell lysates via anion exchange (negative-binding mode), cation exchange, and hydrophobic interaction chromatography, yielding similar to42 and 36 mg EPF from 300 ml bacterial and I L Sf9 cultures, respectively. The preparations were highly purified ( greater than or equal to99% purity on SDS-PAGE for the bacterial products and greater than or equal to97% for that of insect cells) and had the expected mass and heptameric structure under native conditions, as determined by mass spectrometry and gel permeation chromatography, respectively. All recombinant preparations exhibited activity in the EPF bioassay, the rosette inhibition test, with similar potency both to each other and to the native molecule. In two in vivo assays of immuno suppressive activity, the delayed-type hypersensitivity reaction and experimental autoimmune encephalomyelitis, the insect cell and modified bacterial products, both with N-terminal additions (acetylation or amino acid), exhibited similar levels of suppressive activity, but the bacterial product with no N-terminal modification had no effect in either assay. Studies by others have shown that N-terminal addition is not necessary for Cpn10 activity. By defining techniques for facile production of molecules with and without immunosuppressive properties, the present studies make it possible to explore mechanisms underlying the distinction between EPF and Cpn10 activity. (C) 2003 Elsevier Inc. All rights reserved.