Analogs of squalene and oxidosqualene inhibit oxidosqualene cyclase of <Emphasis Type="Italic">Trypanosoma cruzi</Emphasis> expressed in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

Recently, a number of inhibitors of the enzyme oxidosqualene cyclase (OSC; EC 5.4.99.7), a key enzyme in sterol biosynthesis, were shown to inhibit in mammalian cells the multiplication of Trypanosoma cruzi, the parasite agent of Chagas’ disease. The gene coding for the OSC of T. cruzi has been cloned and expressed in Saccharomyces cerevisiae. The expression in yeast cells could be a safe and easy model for studying the activity and the selectivity of the potential inhibitors of T. cruzi OSC. Using a homogenate of S. cerevisiae cells expressing T. cruzi OSC, we have tested 19 inhibitors: aza, methylidene, vinyl sulfide, and conjugated vinyl sulfide derivatives of oxidosqualene and squalene, selected as representative of different classes of substrate analog inhibitors of OSC. The IC₅₀ values of inhibition (the compound concentration at which the enzyme is inhibited by 50%) are compared with the values obtained using OSC of pig liver and S. cerevisiae. Many inhibitors of pig liver and S. cerevisiae OSC show comparable IC₅₀ for T. cruzi OSC, but some phenylthiovinyl derivatives are 10–100 times more effective on the T. cruzi enzyme than on the pig or S. cerevisiae enzymes. The expression of proteins of pathogenic organisms in yeast seems very promising for preliminary screening of compounds that have potential therapeutic activity.     

Synthesis and biological activity of new lodoacetamide derivatives on mutants of squalene-hopene cyclase

New iodoacetamide derivatives, containing a dodecyl or a squalenyl moiety, were synthesized. The effect of these new thiol-reacting molecules was studied on two mutants of Alicyclobacillus acidocaldarius squalene-hopene cyclase constructed especially for this purpose. In the quintuple mutant, all five cysteine residues of the enzyme are substituted with serine; in the sextuple mutant, this quintuple substitution is accompanied by the substitution of aspartate D376, located at the enzyme’s active site, with a cysteine. N-Dodecyliodoacetamide had little activity toward either mutant, whereas N-squalenyliodoacetamide showed a stronger effect on the sextuple than on the quintuple mutant, as expected.     

Presence of the nematode Physaloptera sibirica Petrow and Gorbunow 1931, parasite of carnivores, in the lerot Eliomys quercinus L. in the Alps

Radiolabelled sarcoma cells injected into the tail veins of normal rats were held up almost exclusively in the lung, and were not observed to pass through into the systemic circulation. Intramuscularly injected tumour cells were retained at the site of injection. Radioactivity was lost from both sites though more rapidly from the lung than from muscular tissue and was probably the result of tumour-cell death. Alveolar macrophages did not take part in the destruction of tumour cells in the lung. There was an increased rate of radiolabel loss from the lungs of hyperimmune, post-excision and tumour-bearing rats, as compared with normal rats. The destruction was immunologically specific; it was detected earlier, was more comprehensive in the hyperimmune and post-excision animals than in tumour-bearing animals, and correlated with the ability of the hyperimmune and post-excision animals to reject larger numbers of intravenous unlabelled tumour cells, than the tumour-bearing rats.     

Chemical absorption and desorption of carbon dioxide from hot carbonate solutions

Absorption and desorption rate data for the system CO₂-hot carbonate solutions are presented. The data are interpreted on the basis of a film-theory model developed following the procedure recently presented by Astarita and Savage [1]. The agreement is very satisfactory. Values of the kinetic constant of the rate-determining step, previously known only up to a temperature of 40°C, have been obtained up to 110°C.     

Application of an optimized AM procedure following a SBO in a VVER-1000

The University of Pisa was involved in investigations of an Accident Management (AM) procedure based on passive feed water injection. Some experiments were performed to validate this possibility (e.g. in LOBI and Bethsy facilities) and fully analyzed by thermal hydraulic system codes. Recent activities in which the University of Pisa is engaged (also as leader) are focused on VVER-1000 safety analyses. The idea is now to use the acquired knowledge to explore if a procedure based on passive feed water injection is applicable and can provide any benefits to the Russian design pressurised plant.The postulated accident is a station blackout, in such a way only passive systems are available. The proposed AM is based on secondary and primary side depressurisation in sequence. The secondary side depressurisation performed by the BRU-A valves has the scope to feed passively the SGs with the water left in the feed water lines and in the deaerators. The primary side depressurisation, via the PORV, is foreseen to keep the plant at the lowest pressure (to reduce the energy of the system) and to maximize the “grace time” of the plant. Three cases are here considered: no operator action, application of the optimized AM sequence, application of the AM procedure at the last time when it is effective.The intention of this paper is to show that in case of an unlikely event such a SBO the implementation of a strategy based on systems not designed for specific safety application can have a large impact on the “grace time” of the plant.