Integral equation formulation for buoyancy-driven convection problems

An integral equation formulation for buoyancy-driven convection problems is developed and illustrated. Buoyancy-driven convection in a bounded cylindrical geometry with a free surface is studied for a range of aspect ratios and Nusselt numbers. The critical Rayleigh number, the nature of the cellular motion, and the heat transfer enhancement are computed using linear theory. Green's functions are used to convert the linear problem into linear Fredholm integral equations. Theorems are proved which establish the properties of the eigenvalues and eigenfunctions of the linear integral operator which appears in these equations.     

Viscosity of Strain Gradient Effects on the Kinetics of Propagating Phase Boundaries in Solids

The theory of thermoelastic materials undergoing solid-solid phase transformations requires constitutive information that governs the evolution of a phase boundary. This is known as a kinetic relation which relates a driving traction to the speed of propagation of a phase boundary. The kinetic relation is prescribed in the theory from the onset. Here, though, a special kinetic relation is derived from an augmented theory that includes viscous, strain gradient and heat conduction effects. Based on a special class of solutions, namely travelling waves, the kinetic relation is inherited from the augmented theory as the viscosity, strain gradient and heat conductivity are removed by a suitable limit process.     

Crystalline damage growth during martensitic phase transformations

A thermomechanical model is developed within a large deformation setting in order to simulate the interactions between martensitic phase transformations and crystalline damage growth at the austenitic grain level. Subgrain information is included in the model via the crystallographic theory of martensitic transformations. The damage and transformation characteristics are dependent of the specific martensitic transformation systems activated during a loading process, which makes the model strongly anisotropic. The state of transformation for the individual transformation systems is represented by the corresponding volume fractions. The state of damage in the austenite and in the martensitic transformation systems is reflected by the corresponding damaged volume fractions. The thermodynamical forces energetically conjugated to the rate of volume fraction and the rate of damaged volume fraction are the driving forces for transformation and crystalline damage, respectively. The expressions for these driving forces follow after constructing the specific form of the Helmholtz energy for a phase-changing, damaging material. The model is used to analyze several three-dimensional boundary value problems that are representative of microstructures appearing in multiphase carbon steels containing retained austenite. The analyses show that the incorporation of damage in the model effectively limits the elastic stresses developing in the martensitic product phase, where the maximum value of the stress strongly depends on the toughness of the martensite. Furthermore, in an aggregate of randomly oriented grains of retained austenite embedded in a ferritic matrix the generation of crystalline damage delays the phase transformation process, and may arrest it if the martensitic product phase is sufficiently brittle. The response characteristics computed with the phase-changing damage model are confirmed by experimental results.     

A novel 14C-postlabeling assay using accelerator mass spectrometry for the detection of O6-methyldeoxy-guanosine adducts

Accelerator mass spectrometry (AMS) is currently one of the most sensitive methods available for the trace detection of DNA adducts and is particularly valuable for measuring adducts in humans or animal models. However, the standard approach requires administration of a radiolabeled compound. As an alternative, we have developed a preliminary 14C-postlabeling assay for detection of the highly mutagenic O6-methyldeoxyguanosine (O6-MedG), by AMS. Procedures were developed for derivatising O6-MedG using unlabeled acetic anhydride. Using conventional liquid chromatography/mass spectrometry (LC/MS) analysis, the limit of detection (LOD) for the major product, triacetylated O6-MedG, was 10 fmol. On reaction of O6-MedG with 14C-acetic anhydride, using a specially designed enclosed system, the predominant product was 14C-di-acetyl O6-MedG. This change in reaction profile was due to a modification of the reaction procedure, introduced as a necessary safety precaution. The LOD for 14C-di-acetyl O6-MedG by AMS was determined as 79 amol, approximately 18,000-fold lower than that achievable by liquid scintillation counting (LSC). Although the assay has so far only been carried out with labeled standards, the degree of sensitivity obtained illustrates the potential of this assay for measuring O6-MedG levels in humans.     

Permanently oriented antibody immobilization for digoxin determination with a flow-through fluoroimmunosensor

This paper reports a new flow-through fluoroimmunosensor, the function of which is based on antibodies immobilized on an inmunoreactor of controlled-pore glass (CPG), for determination of digoxin, used in the treatment of congestive heart failure and artery disease. The immunosensor has a detection limit of 1.20 microg L(-1) and provides high reproducibility (RSD=4.5% for a concentration of 0.0025 mg L(-1), and RSD=6.7% for 0.01 mg L(-1)). The optimum working concentration range was found to be 1.2 x 10(-3)-4.0 x 10(-2) mg L(-1). The lifetime of the immunosensor was about 50 immunoassays; if stored unused its lifetime can be extended to three months. A sample speed of about 10-12 samples per hour can be attained. Possible interference from substances with structures similar to digoxin (morphine, heroin, tebaine, codeine, pentazocine and narcotine) was investigated. No cross-reactivity was seen at the highest digoxin: interferent ratio studied (1:100). The proposed fluoroimmunosensor was successfully used to determine digoxin concentrations in human serum samples.     

Single sample extraction protocol for the quantification of NAD and NADH redox states in Saccharomyces cerevisiae

A robust redox extraction protocol for quantitative and reproducible metabolite isolation and recovery has been developed for simultaneous measurement of nicotinamide adenine dinucleotide (NAD) and its reduced form, NADH, from Saccharomyces cerevisiae. Following culture in liquid media, yeast cells were harvested by centrifugation and then lysed under nonoxidizing conditions by bead blasting in ice-cold, nitrogen-saturated 50 mM ammonium acetate. To enable protein denaturation, ice cold nitrogen-saturated CH(3)CN/50 mM ammonium acetate (3:1 v/v) was added to the cell lysates. Chloroform extractions were performed on supernatants to remove organic solvent. Samples were lyophilized and resuspended in 50 mM ammonium acetate. NAD and NADH were separated by HPLC and quantified using UV-Vis absorbance detection. NAD and NADH levels were evaluated in yeast grown under normal (2% glucose) and calorie restricted (0.5% glucose) conditions. Results demonstrate that it is possible to perform a single preparation to reliably and robustly quantitate both NAD and NADH contents in the same sample. Robustness of the protocol suggests it will be (i) applicable to quantification of these metabolites in other cell cultures; and (ii) amenable to isotope labeling strategies to determine the relative contribution of specific metabolic pathways to total NAD and NADH levels in cell cultures.