Direct monitoring of the calcium concentration in the sarcoplasmic and endoplasmic reticulum of skeletal muscle myotubes

Direct monitoring of the free Ca2+ concentration in the sarcoplasmic reticulum (SR) was carried out in rat skeletal myotubes transfected with a specifically targeted aequorin chimera (srAEQ). Myotubes were also transfected with a chimeric aequorin (erAEQ) that we have demonstrated previously is retained in the endoplasmic reticulum (ER). Immunolocalization analysis showed that although both recombinant proteins are distributed in an endomembrane network identifiable with immature SR, the erAEQ protein was retained also in the perinuclear membrane. The difficulty of measuring [Ca2+] in 100-1000 microM range was overcome with the use of the synthetic coelenterazine analogue, coelenterazine n. We demonstrate that the steady state levels of [Ca2+] measured with srAEQ is around 300 microM, whereas that measured with erAEQ is significantly lower, i.e. around 200 microM. The effects of caffeine, high KCl, and nicotinic receptor stimulation, in the presence or absence of external calcium or after blockade of the Ca-ATPase, were investigated with both chimeras. The kinetics of [Ca2+] changes revealed by the erAEQ were similar, but not identical, neither quantitatively nor qualitatively, to those monitored with the srAEQ, indicating that at this stage of muscle development, differences exist between SR and ER in their mechanisms of Ca2+ handling. The functional implications of these findings are discussed.     

Ultrasound‐activated Knoevenagel condensation of malononitrile with carbonylic compounds catalysed by alkaline‐doped saponites

α,β‐Unsaturated nitriles have been synthesized by Knoevenagel condensation of a carbonylic compound with malononitrile, assisted by sonochemical irradiation. Two alkaline‐promoted clays (Li⁺‐ and Cs⁺‐exchanged saponites) have been employed as catalysts. The influence of the carbonylic compound (benzaldehyde or cyclohexanone) and the use of a solvent on the catalytic activity have been studied. Remarkable increase in the conversion values has been found when the reaction is activated by ultrasound, as compared with the thermal activation. In this green, solvent‐free procedure, α,β‐unsaturated nitriles have been produced in very high yields (97%) when the Cs⁺‐saponite is used as catalyst. Copyright © 2004 Society of Chemical Industry     

Nitrification parameter measurement for plant design: experience and experimental issues with new methods.

Nitrification kinetics are important for process design, optimization, and capacity rating of activated sludge wastewater treatment plants. A Water Environment Research Foundation (WERF) project on Methods for Wastewater Characterization in Activated Sludge Modeling (WERF, 2003) focused significantly on the development of procedures for measuring the nitrifier maximum specific growth rate, micro(AUT). In addition, the importance of (and lack of data for) the nitrifier decay rate, b(AUT), was identified. This paper describes three bench-scale methods for measuring micro(AUT): the Low F/M SBR, Washout and High F/M methods. During the WERF project, the importance of pH and temperature control was investigated briefly; this paper summarizes further experimental work performed to address these issues. A summary of micro(AUT) measurements in a number of locations and using the different measurement techniques is provided.     

Performance of a non‐load‐bearing steel stud gypsum board wall assembly: Experiments and modelling

A gypsum wall assembly was exposed to an intense real‐scale compartment fire. For the wall assembly, temperatures were measured at the exposed face, within the stud cavity, and at the unexposed face during the fire exposure. Total heat flux gauges were used to measure the temporal variation of the energy incident on the walls, and cameras, both visual and infrared, were used to image the unexposed face of the wall assembly during the fire exposure. The behaviour of the wall assembly under the fire load is discussed as are current model results for a simulation of the fire test. Copyright © 2006 John Wiley & Sons, Ltd.     

Organochlorine Contaminants of Wintering Ducks Foraging on Detroit River Sediments

Organochlorine analysis was performed on carcasses of 13 diving ducks from a 1981 wintering population that foraged on contaminated sediments in the lower Detroit River. Mean total PCB concentrations were 10 mg/kg for seven lesser scaups (Aythya affinis), 11 mg/kg for three greater scaups (A. marila), and 7.6 mg/kg for three goldeneyes (Bucephala clangula). Highest mean levels of other residues were measured for hexachlorobenzene (1.7 mg/kg) in goldeneyes, and transnonachlor (0.33 mg/kg) and 4,4’-DDE (1.3 mg/kg) in greater scaup. Quantitative analysis of 72 PCB congeners also was applied to water, seston, sediment, benthie oligochaetes, and carp from the same site. Principal congeners in most of the samples included some of the more toxic and persistent PCBs. Results of a multivariate analysis indicated that ratios of more conservative to less conservative PCBs did not vary significantly among ducks (α = .05), but differed from those in carp, oligochaetes, and sediment. For the February through March period of fat mobilization, concentrations of total PCB in lipid were inversely correlated with percent lipid (r = 0.76) in ducks. The percentage of conservative PCBs increased slightly. The distribution and partitioning of organochlorines, including toxic PCB congeners, varied considerably within this water column-sediment-fauna ecosystem. Estimates of toxic exposure based on total PCB values may be unreliable.     

Variations in software development practices

Research on over 18 years of software projects reveals that developing large systems involves substantially more activities and a greater variety of specialized personnel than developing smaller systems. Perhaps the most significant observation is that good quality control is the best overall indicator of a successful project. Schedule delays and cost overruns most often occur when you discover during testing that the application has so many bugs that it doesn't work. Projects using QA teams, formal design and code inspections, and pretest defect tracking always had shorter testing cycles and therefore were more likely to be deliverable on schedule.     

Demonstration of an endurance limit in cast 319 aluminum

An investigation of the fatigue properties of cast W319-T7, an Al-Si-Cu alloy used in automotive engine components, was conducted using ultrasonic testing equipment with operating frequencies of 20 kHz. The stress-life (S-N) behavior at room temperature was determined for three solidification conditions of this alloy, where stresses for fatigue lives ranging from 10⁵ to 10⁹ cycles were determined. The results are compared to fatigue data acquired using servohydraulic equipment operating at 40 Hz. No influence of loading frequency has been observed. A discrete endurance limit is indicated for each of the three solidification conditions of W319-T7. The scientific and practical implications of this result are discussed. A material model presented previously is modified by introducing a crack growth threshold condition in order to predict the observed endurance limits. The model is shown to effectively predict the influence of solidification time on the fatigue properties of W319-T7.