Determination of the biological activity of Clostridium difficile toxins in in vivo and in vitro experiments

The biological activity of the filtrates of 29 C. difficile strains was studied in vivo (suckling white mice) and in vitro (cell cultures of different species and origin). The action of the filtrates on the experimental models in vivo was evaluated from the cytotoxic effect index, while in vitro the intensity of the cytotoxic effect was evaluated from the percentage of dead cells in the monolayer. The results of the comparative determination of toxicity characteristics in vivo and in vitro demonstrated that cell cultures were more sensitive experimental models than suckling white mice. The use of cell cultures permitted the quantitative evaluation of the cytotoxic activity of the filtrates under study, as well as the detection of their cell-directed action at minimal concentrations.     

Molecular mechanism of calcium ion transport in mitochondria. I. Glycoprotein-peptide complex as a component of the electron transport system

The molecular structure of the mitochondrial glycoprotein capable of forming Ca2+-selective and ruthenium red-sensitive conductance channels when incorporating into a model membrane are studied. The glycoprotein is shown to be a complex consisting of the glycoprotein itself and a low-molecular component which may be attributed to the substance of a peptide nature. A technique is elaborated to divide the complex into constituents. It is found that the channel-forming part of the complex is its peptide component. The glycoprotein component is unable to transport Ca2+ and, probably, fulfills a regulatory function.     

Anaerobic degradation of papermill sludge in a two-phase digester containing rumen microorganisms and colonized polyurethane foam

Summary The use of reticulated polyurethane foam as a support material for the immobilization of methanogenic associations and its application to the anaerobic treatment of fine particulate solid wastes was investigated. The colonization of polyurethane support particles in a continuous upflow reactor fed on a mixture of acetate, propionate and butyrate, was both rapid and dense. The combination of rumen microorganisms and colonized support particles in a two-phase digester resulted in an efficient anaerobic decomposition of papermill sludge.     

Reconstitution and partial purification of the glibenclamide-sensitive, ATP-dependent K+ channel from rat liver and beef heart mitochondria.

The transport properties of mitochondria are such that net potassium flux across the inner membrane determines mitochondrial volume. It has been known that K+ uptake is mediated by diffusive leak driven by the high electrical membrane potential maintained by redox-driven, electrogenic proton ejection and that regulated K+ efflux is mediated by an 82-kDa inner membrane K+/H+ antiporter. There is also long-standing suggestive evidence for the existence of an inner membrane protein designed to catalyze electrophoretic K+ uptake into mitochondria. We report reconstitution of a highly purified inner membrane protein fraction from rat liver and beef heart mitochondria that catalyzes electrophoretic K+ flux in liposomes and channel activity in planar lipid bilayers. The unit conductance of the channel at saturating [K+] is about 30 pS. Reconstituted K+ flux is inhibited with high affinity by ATP and ADP in the presence of divalent cations and by glibenclamide in the absence of divalent cations. The mitochondrial ATP-dependent K+ channel is selective for K+, with a Km of 32 mM, and does not transport Na+. K+ transport depends on voltage in a manner consistent with a channel activity that is not voltage-regulated. Thus, the mitochondrial ATP-dependent K+ channel exhibits properties that are remarkably similar to those of the ATP-dependent K+ channels of plasma membranes.     

K+-dependent enzyme activity of H+-ATPase in the E. coli membrane

It was shown that DCCD-sensitive ATPase activity of isolated membranes and preparations of F1F0 only from anaerobically grown E. coli depended on K+ activity. F1F0 include two additional proteins which correspond to the Trk system. The data improve the possibility to form supercomplex (F1F0-Trk) functioning as the H(+)-K(+)-pump.     

Further characterization of recessive suppression in yeast. Isolation of the low-temperature sensitive mutant of Saccharomyces cerevisiae defective in the assembly of 60 S ribosomal subunit

It has been shown that recessive suppressor mutations in the yeast Saccharomyces cerevisiae may cause sensitivity towards low temperatures (very slow growth or lack of growth at 10 degrees C). One of the sup 1 low temperature sensitive (Lts-) mutants, 26-125A-P-2156, was studied in detail. After a prolonged period of incubation (70 h) under restrictive conditions the protein synthesis apparatus in the mutant cells was irreversibly damaged. In addition, Lts- cells incubated under restrictive conditions synthesize unequal amounts of ribosomal subunits, the level of 60 S subunit being reduced. It has been suggested that the recessive suppression is mediated by a mutation in the gene coding for 60 S subunit component, probably a ribosomal protein. The mutation leads simultaneously to a defect in the assembly of 60 S subunit and to low-temperature sensitive growth of the mutant.     

Effect of weak doses of ethyl alcohol on the resistance of the frog sartorius to injurious concentrations of that substance]

A study was made of the effect of preliminary maintenance of m. sartorius of the grass frog in 0.87 M and 1.09 M ethyl alcohol on their resistance to 3.48 M ethyl alcohol. The maintenance of muscle tissue in 0.87 M and 1.09 M ethyl alcohol for periods, ranging from 15 min. to 2 hours, in different series of experiments led to the increase in its resistance to 3.48 M alcohol by 11--24% as compared with the initial one. The value of increase in resistance depended on the concentration of the agent studied, the duration of maintenance, and of the season. A study of non-specificity of the adaptive effect of low alcohol decreased their resistance to 0.12 M NaF by 33.3% (P less than 0.05) The same concentration of ethyl alcohol applied for periods from 15 mintes to 2 hours either caused no change or decreased significantly the resistance of muscle tissue to the temperature 36 degrees C. This effect of decrease in resistance was even more significant when the resistance to 38 degrees C was challenged. The range of individual variability of resistance of muscles to 3.48 M alcohol and 36 degrees showns that its values in the control and in the experiment are similar.