Quantitative affinity chromatography

The objective of this review is to summarize developments in the use of quantitative affinity chromatography to determine equilibrium constants for solute interactions of biological interest. Affinity chromatography is an extremely versatile method for characterizing interactions between dissimilar reactants because the biospecificity incorporated into the design of the affinity matrix ensures applicability of the method regardless of the relative sizes of the two reacting solutes. Adoption of different experimental strategies, such as column chromatography, simple partition equilibrium experiments, solid-phase immunoassay, and biosensor technology, has led to a situation whereby affinity chromatography affords a means of characterizing interactions governed by an extremely broad range of binding affinities-relatively weak interactions (binding constants below 10(3) M-1) through to interactions with binding constants in excess of 10(9) M-1. In addition to its important role in solute separation and purification, affinity chromatography thus also possesses considerable potential for investigating the functional roles of the reactants thereby purified. (C) 2001 Elsevier Science B.V. All rights reserved.

The calibre of the Foramen of Panizza in Crocodylus porosus is variable and under adrenergic control

The foramen of Panizza is located within the outflow tract of the crocodilian heart, between the left and right aortas. It has been suggested that the foremen of Panizza has a variable calibre, which could explain the profound changes in the distribution of flows and pressure profiles recorded in the right and left aortas. We investigated this possibility using a modified in-situ perfused heart preparation in combination with isolated strip preparations from the outflow tract. In the perfused heart preparation, bolus injections of adrenaline increased the resistance in the foramen of Panizza, indicating a decrease in its diameter. Isolated strip preparations from the outflow tract showed a concentration-dependent increase in tension in response to adrenaline, while vasoactive intestinal polypeptide caused a relaxation in adrenaline pre-contracted strip preparations. We propose that an increase in the diameter of the foremen of Panizza may be important during pulmonary to systemic shunts to allow blood to flow from the left to right aorta (reverse foramen flow) in order to supply the carotid and coronary arteries. During non-shunting conditions, a constricted foramen may prevent excess flow from the right to left aorta during diastole.

Mass balance characterisation of Al-7Si-Mg alloy microstructures as a function of solution treatment time

Mass balance calculations were performed to model the effect of solution treatment time on A356 and A357 alloy microstructures. Image analysis and electron probe microanalysis were used to characterise microstructures and confirm model predictions. In as-cast microstructures, up to 8 times more Mg is tied up in the pi-phase than in Mg2Si. The dissolution of pi is accompanied by a corresponding increase in the amount of beta-phase. This causes the rate of pi dissolution to be limited by the rate of beta formation. It is predicted that solution treatments of the order of tens of minutes at 540degreesC produce near-maximum T6 yield strengths, and that Mg contents in excess of 0.52 wt% have no advantage.

Diurnal changes in the photochemical efficiency of the symbiotic dinoflagellates (Dinophyceae) of corals: photoprotection, photoinactivation and the relationship to coral bleaching

The photochemical efficiency of symbiotic dinoflagellates within the tissues of two reef-building corals in response to normal and excess irradiance at wafer temperatures < 30 C were investigated using pulse amplitude modulated (PAM) chlorophyll fluorescence techniques, Dark-adapted F-v/F-m showed clear diurnal changes, decreasing to a low at solar noon and increasing in the afternoon. However, F-v/F-m also drifted downwards at night or in prolonged darkness, and increased rapidly during the early morning twilight. This parameter also increased when the oxygen concentration of the wafer holding the corals was increased. Such changes have not been described previously, and most probably reflect state transition's associated with PQ pool reduction via chlororespiration. These unusual characteristics may be a feature of an endosymbiotic environment, reflective of the well-documented night-time tissue hypoxia that occurs in corals. F-v/F-m decreased to 0.25 in response to full sunlight in shade-acclimated (shade) colonies of Stylophora pistillata, which is considerably lower than in light-acclimated (sun) colonies. In sun colonies, the reversible decrease in F-v/F-m was caused by a lowering of F-m and F-o suggesting photoprotection and no lasting damage. The decrease in F-v/F-m, however, was caused by a decrease in F-m and an increase in F-o in shade colonies suggesting photoinactivation and long-term cumulative photoinhibition. Shade colonies rapidly lost their symbiotic algae (bleached) during exposure to full sunlight. This study is consistent with the hypothesis that excess light leads to chronic damage of symbiotic dinoflagellates and their eventual removal from reef-building corals. It is significant that this can occur with high light conditions alone.

Cold worked Cu-Fe-Cr alloys

The aim of this project was to investigate the properties of copper rich Cu-Fe-Cr alloys for the purpose of developing a new cost effective, high-strength, high-conductivity copper alloy. This paper reports on the influence of cold work. The age hardening response of the Cu-0.7%Cr-2.0%Fe alloy was minimal, but the resistance to softening was superior to that reported for any commercial high-strength, high-conductivity (HSHC) copper alloy with comparable mechanical and electrical properties. For example, an excess of 85% of the original hardness of the 40% cold worked alloy is retained after holding at 700 degreesC for 1 hour, whereas commercial HSHC Cu-Fe-P alloys have been reported to soften significantly after 1 hours exposure at less than 500 degreesC. The Cu-0.7Cr-2.0Fe alloy would therefore be expected to be more suitable for applications with a significant risk of exposure to elevated temperatures. Optical microscope examination of cold worked and aged microstructures confirmed the high resistance to recrystallization for Cu-0.7%Cr-2.0%Fe. The Zener-Smith drag term, predicting the pinning effect of second phase particles on dislocations in cold worked microstructures, was calculated using the precipitate characteristics obtained from TEM, WDS and resistivity measurements. The pinning effect of the precipitate dispersions in the peak-aged condition was determined to be essentially equivalent for the Cu-0.7%Cr-0.3%Fe and Cu-0.7%Cr-2.0%Fe alloys. A lower recrystallisation temperature in the Cu-0.7%Cr-0.3%Fe alloy was therefore attributed to faster coarsening kinetics of the secondary precipitates resulting from a higher Cr concentration in the precipitates at lower iron content. (C) 2001 Kluwer Academic Publishers.

Determination of breeding strategies for beef cattle bull breeders selling bulls into two competing markets on a non-linear price grid

Computer simulation was used to suggest potential selection strategies for beef cattle breeders with different mixes of clients between two potential markets. The traditional market paid on the basis of carcass weight (CWT), while a new market considered marbling grade in addition to CWT as a basis for payment. Both markets instituted discounts for CWT in excess of 340 kg and light carcasses below 300 kg. Herds were simulated for each price category on the carcass weight grid for the new market. This enabled the establishment of phenotypic relationships among the traits examined [CWT, percent intramuscular fat (IMF), carcass value in the traditional market, carcass value in the new market, and the expected proportion of progeny in elite price cells in the new market pricing grid]. The appropriateness of breeding goals was assessed on the basis of client satisfaction. Satisfaction was determined by the equitable distribution of available stock between markets combined with the assessment of the utility of the animal within the market to which it was assigned. The best goal for breeders with predominantly traditional clients was a CWT in excess of 330 kg, while that for breeders with predominantly new market clients was a CWT of between 310 and 329 kg and with a marbling grade of AAA in the Ontario carcass pricing system. For breeders who wished to satisfy both new and traditional clients, the optimal CWT was 310-329 kg and the optimal marbling grade was AA-AAA. This combination resulted in satisfaction levels of greater than 75% among clients, regardless of the distribution of the clients between the traditional and new marketplaces.

Changes in progesterone and testosterone during the breeding season of the large-footed myotis Myotis moluccarum (Microchiroptera : Vespertilionidae)

The reproductive biology of the large-footed myotis, Myotis moluccarum, was studied during the annual breeding season in southeast Queensland, Australia. Previous research has shown the species to be polyoestrous and monotoccous, producing two consecutive young with some degree of synchrony in late October to early November and again in late January to early February. Hormonal data was collected and observations of the female reproductive tract made in order to ascertain the reproductive cycle of this species. In July, when females were not pregnant, progesterone concentrations were 1.9 +/- 0.9 ng/ml. During the two gestation periods, progesterone concentrations increased progressively until late pregnancy at the end of October through to early November and again in late January to early February. During the latest stages of pregnancy, progesterone concentrations of 69.9 +/- 18.7 ng/ml were reached. It is suggested that a plasma progesterone concentration in excess of about 8 ng/ml indicates pregnancy in this species. Plasma testosterone concentration in males reached a peak of 43.1 +/- 9.81 ng/ml in July, and was then variable until December when levels declined significantly to 2.0 +/- 1.7 ng/ml.

Operating space diagrams: a tool for designers of wastewater treatment plants

Recent years have seen the introduction of new and varied designs of activated sludge plants. With increasing needs for higher efficiencies and lower costs, the possibility of a plant that operates more effectively has created the need for tools that can be used to evaluate and compare designs at the design stage. One such tool is the operating space diagram. It is the aim of this paper to present this tool and demonstrate its application and relevance to design using a simple case study. In the case study, use of the operating space diagram suggested changes in design that would improve the flexibility of the process. It also was useful for designing suitable control strategies.

A multi-purpose rainfall simulator for field infiltration and erosion studies

This paper describes a rainfall simulator developed for field and laboratory studies that gives great flexibility in plot size covered, that is highly portable and able to be used on steep slopes, and that is economical in its water use. The simulator uses Veejet 80100 nozzles mounted on a manifold, with the nozzles controlled to sweep to and from across a plot width of 1.5 m. Effective rainfall intensity is controlled by the frequency with which the nozzles sweep. Spatial uniformity of rainfall on the plots is high, with coefficients of variation (CV) on the body of the plot being 8-10%. Use of the simulator for erosion and infiltration measurements is discussed.

Clinical trials with glycoprotein IIB/IIIA antagonists - No benefit without bleeding?

As the glycoprotein GPIIb/IIIa receptor is the final common pathway in platelet aggregation, antagonists of this receptor cause a profound inhibition of aggregation induced by any agonist. The short-term efficacy and safety of GPIIb/IIIa antagonists in patients undergoing coronary angioplasty was demonstrated with murine 7E3 Fab, but this antibody was immunogenic. Abciximab is a chimeric human-mouse monoclonal antibody that is less immunogenic. The first major trial with a GPIIb/IIIa antagonist was the EPIC trial with abciximab, which showed that abciximab reduced the ischemic complications of coronary balloon angioplasty and atherectomy in high-risk patients, but increased the risk of bleeding. Subsequent studies showed that using less concurrent heparin reduced bleeding. Abciximab also reduced the rate of revascularization. Further studies have shown that the benefits of abciximab extended to all patients undergoing angioplasty (EPILOG), including patients with unstable angina (CAPTURE) and acute myocardial infarction (RAPPORT). Clinical trials with eptifibatide and tirofiban have failed to demonstrate benefit, at the doses used, in angioplasty. Abciximab and eptifibatide, but not oral xemilofiban, improve the safety of the coronary stenting procedure. Shortterm intravenous treatment with lamifiban, eptifibatide or tirofiban is beneficial in acute coronary syndromes (unstable angina, non-Q wave myocardial infarction). Orally active GPIIb/IIIa antagonists are being developed for use in acute coronary syndromes and myocardial infarction. However, no benefit has been shown with lefradafiban in acute coronary syndromes and sibrafiban and orbofiban are harmful. Eptifibatide, lamifiban and abciximab improve coronary patency in myocardial infarction, and long-term trials of GPIIb/IIIa antagonists are being conducted in acute myocardial infarction. Abciximab can cause thrombocytopenia, and all the GPIIb/IIIa antagonists increase the incidence of bleeding, but there is no excess of intracranial hemorrhage. (C) 2001 Prous Science. All rights reserved.

Modern scientific methods and their potential in wastewater science and technology

Application of novel analytical and investigative methods such as fluorescence in situ hybridization, confocal laser scanning microscopy (CLSM), microelectrodes and advanced numerical simulation has led to new insights into micro-and macroscopic processes in bioreactors. However, the question is still open whether or not these new findings and the subsequent gain of knowledge are of significant practical relevance and if so, where and how. To find suitable answers it is necessary for engineers to know what can be expected by applying these modern analytical tools. Similarly, scientists could benefit significantly from an intensive dialogue with engineers in order to find out about practical problems and conditions existing in wastewater treatment systems. In this paper, an attempt is made to help bridge the gap between science and engineering in biological wastewater treatment. We provide an overview of recently developed methods in microbiology and in mathematical modeling and numerical simulation. A questionnaire is presented which may help generate a platform from which further technical and scientific developments can be accomplished. Both the paper and the questionnaire are aimed at encouraging scientists and engineers to enter into an intensive, mutually beneficial dialogue. (C) 2002 Elsevier Science Ltd. All rights reserved.

Enrichment of autotrophic anaerobic ammonium-oxidizing consortia from various wastewaters

The option for biological nitrogen removal has recently been broadened with the description of simultaneous nitrification/denitrification, anaerobic ammonium oxidation (ANAMMOX) and the concept of CANON (completely autotrophic nitrogen removal over nitrite). An autotrophic anaerobic ammonium oxidation (AAAO) consortium was successfully selected and enriched from municipal treatment plant sludges in Sydney, Australia, but not from industrial coke-oven wastewater sludges. Chemolithoautotrophic basic salt (CLABS) medium was used in the selection of AAAO organisms and chloramphenicol was added to the initial stage of selection to eliminate denitrifiers. Two different temperatures, 37degreesC and 55degreesC, were used in the selection of mesophilic and thermophilic consortia, respectively. Thermophilic AAAO organisms were not selected at 55degreesC. Mesophilic AAAO activities, however, were evident in both batch and continuous cultures, whereby ammonium was consumed concurrently with a decrease of nitrite, giving a ratio of 1:1-1:1.3 in ammonium removal rate over nitrite consumption rate. A continuous-mode mesophilic fixed-bed reactor was established to enrich the AAAO consortium. After 1 year, biofilms, pinkish in color, had developed on the support media and side wall of the feed-line tubing. Ammonium and nitrite consumption increased from similar to15 mg to 60 mg d(-1) L-1 over a period of 243 days. Later, transmission electron microscopy (TEM) and fluorescence in situ hybridization (FISH) techniques revealed that the dominant cell type in the AAAO consortium had a similar morphology and 16S rDNA sequence homology to that of the recently described ANAMMOX organism, Brocadia anammoxidans.

Interferon-gamma enhances cytotoxic T lymphocyte recognition of endogenous peptide in keratinocytes without lowering the requirement for surface peptide

Keratinocytes expressing the human papillomavirus (HPV) type 16 E7 protein, as a transgene driven by the K14 promoter, form a murine model of HPV-mediated epithelial cancers in humans. Our previous studies have shown that K14E7 transgenic skin grafts onto syngeneic mice are not susceptible to immune destruction despite the demonstrated presence of a strong, systemic CTL response directed against the E7 protein. Consistent with this finding, we now show that cultured, E7 transgenic keratinocytes (KC) express comparable endogenous levels of E7 protein to a range of CTL-sensitive E7-expressing cell lines but are not susceptible to CTL-mediated lysis in vitro . E7 transgenic and non-transgenic KC are susceptible to conventional mechanisms of CTL-mediated lysis, including perforin and Fas/FasL interaction when an excess of exogenous peptide is provided. The concentration of exogenous peptide required to render a cell susceptible to lysis was similar between KC and other conventional CTL targets (e.g. EL-4), despite large differences in H-2D(b) expression at the cell surface. Furthermore, exposure of KC to IFN-gamma increased H-2D(b) expression, but did not substantially alter the exogenous peptide concentration required to sensitize cells for half maximal lysis. In contrast, the lytic sensitivity of transgenic KC expressing endogenous E7 is modestly improved by exposure to IFN-gamma. Thus, failure of CTL to eliminate KC expressing endogenous E7, and by inference squamous tumours expressing E7, may reflect the need for a sustained, local inflammatory environment during the immune effector phase.

Analysis of the microbial community structure and function of a laboratory scale enhanced biological phosphorus removal reactor

A laboratory scale sequencing batch reactor (SBR) operating for enhanced biological phosphorus removal (EBPR) and fed with a mixture of volatile fatty acids (VFAs) showed stable and efficient EBPR capacity over a four-year-period. Phosphorus (P), poly-beta-hydroxyalkanoate (PHA) and glycogen cycling consistent with classical anaerobic/aerobic EBPR were demonstrated with the order of anaerobic VFA uptake being propionate, acetate then butyrate. The SBR was operated without pH control and 63.67+/-13.86 mg P l(-1) was released anaerobically. The P% of the sludge fluctuated between 6% and 10% over the operating period (average of 8.04+/-1.31%). Four main morphological types of floc-forming bacteria were observed in the sludge during one year of in-tensive microscopic observation. Two of them were mainly responsible for anaerobic/aerobic P and PHA transformations. Fluorescence in situ hybridization (FISH) and post-FISH chemical staining for intracellular polyphosphate and PHA were used to determine that 'Candidatus Accumulibacter phosphatis' was the most abundant polyphosphate accumulating organism (PAO), forming large clusters of coccobacilli (1.0-1.5 mum) and comprising 53% of the sludge bacteria. Also by these methods, large coccobacillus-shaped gammaproteobacteria (2.5-3.5 mum) from a recently described novel cluster were glycogen-accumulating organisms (GAOs) comprising 13% of the bacteria. Tetrad-forming organisms (TFOs) consistent with the 'G bacterium' morphotype were alphaproteobacteria , but not Amaricoccus spp., and comprised 25% of all bacteria. According to chemical staining, TFOs were occasionally able to store PHA anaerobically and utilize it aerobically.

Field quantization, photons and non-Hermitean modes

Field quantization in unstable optical systems is treated by expanding the vector potential in terms of non-Hermitean (Fox-Li) modes. We define non-Hermitean modes and their adjoints in both the cavity and external regions and make use of the important bi-orthogonality relationships that exist within each mode set. We employ a standard canonical quantization procedure involving the introduction of generalized coordinates and momenta for the electromagnetic (EM) field. Three-dimensional systems are treated, making use of the paraxial and monochromaticity approximations for the cavity non-Hermitean modes. We show that the quantum EM field is equivalent to a set of quantum harmonic oscillators (QHOs), associated with either the cavity or the external region non-Hermitean modes, and thus confirming the validity of the photon model in unstable optical systems. Unlike in the conventional (Hermitean mode) case, the annihilation and creation operators we define for each QHO are not Hermitean adjoints. It is shown that the quantum Hamiltonian for the EM field is the sum of non-commuting cavity and external region contributions, each of which can be expressed as a sum of independent QHO Hamiltonians for each non-Hermitean mode, except that the external field Hamiltonian also includes a coupling term responsible for external non-Hermitean mode photon exchange processes. The non-commutativity of certain cavity and external region annihilation and creation operators is associated with cavity energy gain and loss processes, and may be described in terms of surface integrals involving cavity and external region non-Hermitean mode functions on the cavity-external region boundary. Using the essential states approach and the rotating wave approximation, our results are applied to the spontaneous decay of a two-level atom inside an unstable cavity. We find that atomic transitions leading to cavity non-Hermitean mode photon absorption are associated with a different coupling constant to that for transitions leading to photon emission, a feature consequent on the use of non-Hermitean mode functions. We show that under certain conditions the spontaneous decay rate is enhanced by the Petermann factor.