A genome-wide screen identifies yeast genes required for protection against or enhanced cytotoxicity of the antimalarial drug quinine

Quinine is used in the treatment of Plasmodium falciparum severe malaria. However, both the drug's mode of action and mechanisms of resistance are still poorly understood and subject to debate. In an effort to clarify these questions, we used the yeast Saccharomyces cerevisiae as a model for pharmacological studies with quinine. Following on a previous work that examined the yeast genomic expression program in response to quinine, we now explore a genome-wide screen for altered susceptibility to quinine using the EUROSCARF collection of yeast deletion strains. We identified 279 quinine-susceptible strains, among which 112 conferred a hyper-susceptibility phenotype. The expression of these genes, mainly involved in carbohydrate metabolism, iron uptake and ion homeostasis functions, is required for quinine resistance in yeast. Sixty-two genes whose deletion leads to increased quinine resistance were also identified in this screen, including several genes encoding ribosome protein subunits. These well-known potential drug targets in Plasmodium are associated with quinine action for the first time in this study. The suggested involvement of phosphate signaling and transport in quinine tolerance was also studied, and activation of phosphate starvation-responsive genes was observed under a mild-induced quinine stress. Finally, P. falciparum homology searches were performed for a selected group of 41 genes. Thirty-two encoded proteins possess homologs in the parasite, including subunits of a parasitic vacuolar H(+)-ATPase complex, ion and phosphate importers, and several ribosome protein subunits, suggesting that the results obtained in yeast are good candidates to be transposed and explored in a P. falciparum context.     

Spirograph for small laboratory animals

A design of dry spirograph is described. It is characterized by greater precision, lack of inertia, high reliability, absence of respiration resistance, adequate form of recording, rapid resetting to any respiratory rate. The device consists of two similar injection syringes, photoelectric sensor for the identification of the initial moments of respiration stages, electromagnetic valves, two photoelectric converters of the air volume into the impulse signal, vacuum micro-pump, microcompressor and a system of air-driving tubes. In the initial position of pistons and valves the microcompressor pumps air into the inhalation cylinder and lifts the piston to the upper extreme position. With the signal marking the beginning of inspiration, the valves switch over and the piston lowers, pushing out the air, which moves into the animals' respiratory organs. Simultaneously, the signals of the inhaled air volume from the photoelectric transducer reach the recorder. During expiration the air pushes the piston down into the second cylinder and photoelectric transducer gives the information on the volume of the expired air.     

Evidence for protection by heat-shock proteins against photoinhibition during heat-shock

The nuclear-coded 22 kd heat-shock protein (HSP-22) which is transported into the chloroplast and localized in the thylakoids was further characterized and found to be located in the grana lamellae (stacked thylakoids) as an extrinsic protein in the green alga Chlamydomonas reinhardtii. Inhibition of photosynthetic electron flow during heat-shock of Chlamydomonas cells was light-dependent, occurring at low-light intensities (<100 W/m²) as compared with photoinhibition at 25°C (>1000 W/m²). The site of the damage was localized at the photosystem II (PS II) reaction center. The damage was drastically increased when heat-shock treatment was carried out in the presence of the 80S ribosomal translation inhibitor, cycloheximide (CHI). Pre-incubation of Chlamydomonas cells at 42°C resulted in partial protection against photoinhibition during heat-shock, as compared with cells pre-incubated at 42°C in the presence of CHI which, therefore, did not translate the heat-shock proteins. Analysis of the thylakoid polypeptides' pattern by SDS-PAGE revealed that during heat-shock in the light, thylakoid proteins became aggregated proportionally to the light intensity. Heat-shock in the presence of CHI enhanced the aggregation process which, at low light intensities, was specific to the PS II reaction center D1-protein. The results suggest that the chloroplasts HSPs prevent damage to the PS II reaction center during heat-shock in the light.     

Ion exposure chambers for small animals

Ion exposure chambers that have been designed and tested for use in biological and behavioral research with small animals are described in this report. The chambers exhibit an acceptable degree of uniformity in ion concentration, current density, and electric field within the exposure area. Gaseous by-products of corona discharge (O3 and NO2) have been measured and found to be less than .01 ppm and less than .1 ppm, respectively. Filtered air is fed to the individual exposure chambers, and temperature and humidity are well controlled. Noise due to corona and the air delivery system has been measured.