Triangle Distribution and Equation of State for Classical Rigid Disks

The triangle distribution function f ⁽³⁾ for three mutual near neighbors in the plane describes basic aspects of short-range order and statistical thermodynamics in two-dimensional many-particle systems. This paper examines prospects for constructing a self-consistent calculation for the rigid-disk-system f ⁽³⁾. We present several identities obeyed by f ⁽³⁾. A rudimentary closure suggested by scaled-particle theory is introduced. In conjunction with three of the basic identities, this closure leads to an unique f ⁽³⁾ over the entire density range. The pressure equation of state exhibits qualitatively correct behaviors in both the low-density and the close-packed limits, but no intervening phase transition appears. We discuss extensions to improved disk closures, and to the three-dimensional rigid-sphere system.     

Continuous dynamic-gravimetric preparation of calibration gas mixtures for air pollution measurements

 The preparation of calibration gas mixtures for air pollution measurements by the dynamic-gravimetric method was investigated using sulphur dioxide in nitrogen as a model. The target mole fraction was 200×10^–9 mol/mol, with the option of also getting smaller mole fractions. Thermal mass flow meters calibrated with reference mass flows were used to measure the dilution gas flow (nitrogen). The relative standard uncertainty of the dilution gas flows between 10 mg/s (approx. 500 ml/min) and 40 mg/s (approx. 2000 ml/min) was 0.15%. The mass flow of the target component measured as the permeation rate was determined via the quasi-continuous observation of the loss in the permeation tube mass during the measuring time. A magnetic coupling system and an adapted microbalance were used for this purpose. The results presented show permeation rates measured over the lifetime of a tubular permeation source. The measurement cycles took between 3 days and 7 h at least. The relative standard uncertainty of the mixture composition did not exceed 2%. First comparisons with gas mixtures prepared by the static-gravimetric method show compatibility. The applicability of the system is not restricted to the SO₂/N₂ mixture. It can also be used for preparing other gas mixtures in this field of application. Received: 26 April 2000 / Accepted: 12 September 2000     

Intracellularly grown gold nanoislands as SERS substrates for monitoring chromate,sulfate and nitrate localization sites in remediating bacteria biofilms by Raman chemical imaging

Understanding the chemical composition of biofilm matrices is vital in different fields of biology such as surgery, dental medicine, synthetic grafts and bioremediation. The knowledge of biofilm development, composition, active reduction sites and remediation efficacy will help in the development of effective solutions and evaluation of remediating approaches prior to implementation. Surface-enhanced Raman spectroscopy (SERS) based imaging is an invaluable tool to obtain an understanding of the remediating efficacy of microorganisms and its role in the formation of organic and inorganic compounds in biofilms. We demonstrate for the first time, the presence of chromate, sulfate, nitrate and reduced trivalent chromium in soil biofilms. In addition, we demonstrate that SERS imaging was able to validate two observations made by previous studies on chromate/sulfate and chromate/nitrate interactions in Shewanella oneidensis MR-1 biofilms. Additionally, we show a detailed Raman mapping based evidence of the existence of chromate–sulfate competition for cellular entry. Subsequently, we use Raman mapping to study the effect of nitrate on chromate reduction. The findings presented in this paper are among the first to report – detection of multiple metallic ions in bacterial biofilms using intracellular SERS substrates. Such a detailed characterization of biofilms using gold nanoislands based SERS mapping substrate can be extended to study cellular localization of other metallic ions and chemical species of biological and toxicological significance and their effect on reduction reactions in bacterial biofilms.     

Challenges and trends in the determination of selected chemical contaminants and allergens in food

This article covers challenges and trends in the determination of some major food chemical contaminants and allergens, which-among others-are being monitored by Health Canada's Food Directorate and for which background levels in food and human exposure are being analyzed and calculated. Eleven different contaminants/contaminant groups and allergens have been selected for detailed discussion in this paper. They occur in foods as a result of: use as a food additive or ingredient; processing-induced reactions; food packaging migration; deliberate adulteration; and/or presence as a chemical contaminant or natural toxin in the environment. Examples include acrylamide as a food-processing-induced contaminant, bisphenol A as a food packaging-derived chemical, melamine and related compounds as food adulterants and persistent organic pollutants, and perchlorate as an environmental contaminant. Ochratoxin A, fumonisins, and paralytic shellfish poisoning toxins are examples of naturally occurring toxins whereas sulfites, peanuts, and milk exemplify common allergenic food additives/ingredients. To deal with the increasing number of sample matrices and analytes of interest, two analytical approaches have become increasingly prevalent. The first has been the development of rapid screening methods for a variety of analytes based on immunochemical techniques, utilizing ELISA or surface plasmon resonance technology. The second is the development of highly sophisticated multi-analyte methods based on liquid chromatography coupled with multiple-stage mass spectrometry for identification and simultaneous quantification of a wide range of contaminants, often with much less requirement for tedious cleanup procedures. Whereas rapid screening methods enable testing of large numbers of samples, the multi analyte mass spectrometric methods enable full quantification with confirmation of the analytes of interest. Both approaches are useful when gathering surveillance data to determine occurrence and background levels of both recognized and newly identified contaminants in foods in order to estimate human daily intake for health risk assessment.     

A note on synchronous egg laying in a seabird behaviour model

ABSTRACT

During years when sea surface temperature (SST) is high, gulls in a colony on Protection Island, Washington, USA typically experience low food availability. As SST rises, feeder fish follow plankton to cooler temperatures in deeper water levels. Since gulls are surface-feeding birds, they face a food shortage. A tactic male gulls employ to deal with this food shortage is to cannibalize their neighbours' eggs. Gulls in this colony exhibit an adaptive tactic of every-other-day egg-laying synchrony in response to egg cannibalism, and the level of synchrony increases with colony density. Here we analyze the dynamics of an animal behaviour model for egg laying as a function of colony density. As colony density increases, the equilibrium loses stability in a 2-cycle bifurcation. The 2-cycle becomes increasingly synchronous as the colony density continues to increase. We show that egg-laying synchrony benefits the colony in the presence of cannibalism.     

Comparison of G-protein coupled receptor desensitization-related beta-arrestin redistribution using confocal and non-confocal imaging

High Content Screening (HCS), a combination of fluorescence microscopic imaging and automated image analysis, has become a frequently applied tool to study test compound effects in cellular disease-modelling systems. In this work, we compared a confocal and a non-confocal cellular HCS system, the IN Cell Analyzers(1) 3,000 and 1,000, respectively. As a cellular model system we used the Transfluor technology in the 384-well microtiter plate (MTP) format. The Transfluor HCS assay for G-protein coupled receptor (GPCR) activation is based on the recruitment of a green fluorescent protein-labelled arrestin (ArrGFP) from the cytosol to the plasma membrane. We investigated two GPCRs, the wild-type (wt) beta2 adrenergic receptor (beta2AR) and the beta2AR-enhanced (E), a C-terminally mutated receptor with a higher affinity to arrestin. Upon agonist stimulation, the beta2AR-wt induced the redistribution of ArrGFP to coated pits, the beta2AR-E maintained the interaction with ArrGFP down to the formation of endocytic vesicles. Our findings reveal that the assay is feasible on both instruments, with sufficiently robust Z' statistics. Improved Z' statistics, though, are achieved with the confocal system, particularly in case of weak signals. Moreover, throughput is dramatically higher for the IN Cell Analyzer 3,000. We conclude that, depending on the needs for throughput and assay biology, either instrument may fulfil a successful role in the drug discovery process. Confocal optics, however, provide a better basis for the detection of smaller subcellular structures with lower fluorescence intensity.     

Automated high content screening for phosphoinositide 3 kinase inhibition using an AKT 1 redistribution assay

High Content Screening (HCS), a combination of fluorescence microscopic imaging and automated image analysis, has become a frequently applied tool to study test compound effects in cellular disease-modelling systems. In this work, we established a medium to high throughput HCS assay in the 384-well format to measure cellular type I phosphoinositide 3 kinase (PI3K) activity. Type I PI3K is involved in several intracellular pathways such as cell survival, growth and differentiation as well as immunological responses. As a cellular model system we used Chinese Hamster Ovary (CHO) cells that had been stably transfected with human insulin receptor (hIR) and an AKT1-enhanced green fluorescent protein (EGFP) fusion construct. Upon stimulation of the hIR with insulin-like growth factor-1 (IGF-1), PI3K was activated to phosphorylate phosphatidylinositol (PtdIns)-4,5-bisphosphate at the 3-position, resulting in the recruitment of AKT1-EGFP to the plasma membrane. The AKT1-EGFP redistribution assay was robust and displayed little day-to-day variability, the quantification of the fluorescence intensity associated with plasma membrane spots delivered good Z' statistics. A novel format of compound dose-response testing was employed using serial dilutions of test compounds across consecutive microtiter plates (MTPs). The dose response testing of a PI3K inhibitor series provided reproducible IC50 values. The profiling of the redistribution assay with isoform-selective inhibitors indicates that PI3Kalpha is the main isoform activated in the CHO host cells after IGF-1 stimulation. Toxic compound side effects could be determined using automated image analysis. We conclude that the AKT1-EGFP redistribution assay represents a solid medium/high throughput screening (MTS/HTS) format to determine the cellular activity of PI3K inhibitors under conditions of growth factor stimulation.