A novel copper-regulated promoter system for expression of heterologous proteins in Schizosaccharomyces pombe.

The increasing use of the fission yeast Schizosaccharomyces pombe as a model organism for elucidating the mechanisms of critical biological processes such as cell-cycle control, DNA replication, and stress-mediated signal transduction has fostered the development and utilization of expression systems for gene function analysis. Using the promoter of the ctr4(+) copper transporter gene from S. pombe, we created a series of vectors, named pctr4(+)-X, which regulate the expression of heterologous genes as a function of copper availability. In this system, the addition of copper ions at levels that are non-toxic to yeast cells represses gene expression, while copper deprivation strongly induces gene expression. Conveniently, changes of growth medium or carbon sources are not required to shut down or induce gene expression. The Cu-starvation-mediated inducible expression system is rapid, producing heterologous proteins within 3 h, with sustained expression of proteins that persists for several hours. The pctr4(+)-X expression vectors harbor unique restriction sites constructed in-frame to DNA sequences encoding for epitope tags, which facilitate the detection or purification of the heterologous proteins using commercially available antibodies and affinity columns. Furthermore, the pctr4(+)-X copper-regulatable protein expression vectors have been constructed with three different selectable markers, offering more versatility for studying gene function in fission yeast.

Novel C-terminus modifications of the Dmt-Tic motif: a new class of dipeptide analogues showing altered pharmacological profiles toward the opioid receptors.

The design, synthesis and pharmacological evaluation of a novel class of Dmt-Tic dipeptide analogues are described. These resulting analogues bearing different C-terminal functionalities were found to bind to the human delta receptor with high affinity. One specific class of dipeptides bearing urea/thiourea functionalities showed partial to full activation of the delta receptor. Several dipeptides also showed good binding affinities with full activation of the human kappa receptor, a novel property for those ligands.

Air quality monitoring during indoor Monster Truck and car demolition shows.

This article describes the results of air quality monitoring in an indoor ice skating rink during three Monster Truck and car demolition exhibitions, and the public health study that was carried out. The exposure of the people present to carbon monoxide and nitrogen dioxide was continuously monitored in order to determine the time-weighted average concentrations and the maximum peaks. Nitrogen dioxide concentrations were generally under the limit of detection of the device (0.5 ppm). However, carbon monoxide levels exceeded standards for workers. Maximum time-weighted average concentrations during the exhibitions were 100 parts per million with several peaks exceeding 200 parts per million (maximum value: 1600 parts per million). Recommendations were made and during a subsequent event, the carbon monoxide concentrations were reduced to protect health. Indoor exhibitions of motorized vehicles generate significant amounts of combustion gases, which can be a health hazard. There must be sufficient ventilation and the carbon monoxide and nitrogen dioxide concentrations must be monitored. In addition, the motors of the most polluting vehicles should be adjusted before the events in order to limit the emission of combustion products. If these steps are not met, the events should be held outdoors.

An unusual case of carbon monoxide poisoning.

Carbon monoxide, a gas originating from incomplete combustion of carbon-based fuels, is an important cause of human deaths. In this paper, we describe an unusual carbon monoxide poisoning in a dwelling without obvious sources of combustion gases, for which two adults had to be treated in a hyperbaric chamber. Carbon monoxide readings were taken in the house and in the neighboring homes. Methane gas and nitrogen oxide levels were also monitored in the house air. Soil samples were collected around the house and tested for hydrocarbon residues. The investigation revealed the presence of a pocket of carbon monoxide under the foundation of the house. The first readings revealed carbon monoxide levels of 500 ppm in the basement. The contamination lasted for a week. The investigation indicated that the probable source of contamination was the use of explosives at a nearby rain sewer construction site. The use of explosives in a residential area can constitute a major source of carbon monoxide for the neighboring populations. This must be investigated, and public health authorities, primary-care physicians, governmental authorities, and users and manufacturers of explosives must be made aware of this problem.