Mechanism of action of N-acetylglucosaminyl-N-acetylmuramylalanyl-D-isoglutamine on the nonspecific anti-infection resistance of mice

The stimulating influence of glucose-containing muramyldipeptide (GMDP) on the nonspecific resistance of mice was shown to depend on the features of the pathogenesis of the infection. Thus, the intraperitoneal injection of GMDP increased the survival rate of mice infected with Escherichia coli, but had no stimulating effect on the resistance of the animals to Salmonella typhimurium natural infection in whose pathogenesis macrophages played an essential role. Experiments demonstrated that GMDP was capable of enhancing the ingestive function of macrophages, but did not increase their bactericidal activity with respect to this infection.     

Primary and secondary immune response of mice to polysaccharide antigens of meningococcal serogroups A and C

The peculiarities of the primary immune response, the formation of immunological memory and the secondary immune response to serogroup A and C meningococcal polysaccharides were studied in 7 strains of inbred mice, hybrids F1 and in noninbred animals. The passive local hemolysis test and the passive hemagglutination test indicated that the intensity of immune response to A and C polysacchardies depended on the genotype of the animals: both antigens induced the most intense response in CBA and BALB/c mice. The primary immune response to the both antigens was characterized by a short latent period, a rapid (by days 4-5) increase in the amount of antibody-producing cells in the spleen and in antibody titer in the blood serum to the maximum level, and a pronounced decrease inantibody formation by days 6-7 followed by a gradual extinction of the response. A single injection of A and C polysaccharides in a dose of 0.5 microgram induced the formation of immunological memory in mice, persisting for at least 4 weeks and manifesting after reimmunization as the increased or more prolonged synthesis of IgM and IgG.     

Growth of human embryonic fibroblasts at clonal density: concordance with results from mass cultures.

In the past, it has been difficult to grow human diploid fibroblast cells at clonal densities. Newly devised cell culture media and rigorously controlled environmental conditions have greatly increased the ease with which such cells can be cloned. The present work was undertaken to determine whether, under appropriate conditions, diploid fibroblast cells from human embryonic lung, grow as well at clonal densities as in mass culture. The parameters studied were: (1) population doubling time, (2) in vitro proliferative capacity, (3) attachment, (4) percentage of non-dividing cells. In all cases essentially the same results were obtained for cultures at clonal densities and mass cultures. These results indicate that the behavior of these types of cells in clonal culture can be used to infer the behavior of individual cells and clones within a mass culture.     

High molecular substances in relation to disorders of erythropoiesis in patients in the terminal stage of chronic renal failure

Fraction F1 with a MW of 5.10(3)-5.10(4), which had been isolated from the blood serum of blood donors as controls and patients at the terminal stage of chronic kidney insufficiency, specifically inhibited the erythropoiesis of test animals at the stage sensitive to erythropoietin and morphologically distinguishable cells of the bone-marrow. In uremia, protein concentration was twice higher in comparison to the normal one. The inhibiting effect of fraction F1 from the serum of patients is caused by the enlargement of level of physiological inhibitors and unspecific toxins of the cell proliferation. Those data characterising the factor detected in the fraction F1 of normal serum together with the erythrocytic chalon are discussed.     

Mechanism of maltal hydration catalyzed by beta-amylase: role of protein structure in controlling the steric outcome of reactions catalyzed by a glycosylase.

Crystalline (monomeric) soybean and (tetrameric) sweet potato beta-amylase were shown to catalyze the cis hydration of maltal (alpha-D-glucopyranosyl-2-deoxy-D-arabino-hex-1-enitol) to form beta-2-deoxymaltose. As reported earlier with the sweet potato enzyme, maltal hydration in D2O by soybean beta-amylase was found to exhibit an unusually large solvent deuterium kinetic isotope effect (VH/VD = 6.5), a reaction rate linearly dependent on the mole fraction of deuterium, and 2-deoxy-[2(a)-2H]maltose as product. These results indicate (for each beta-amylase) that protonation is the rate-limiting step in a reaction involving a nearly symmetric one-proton transition state and that maltal is specifically protonated from above the double bond. This is a different stereochemistry than reported for starch hydrolysis. With the hydration catalyzed in H2O and analyzed by gas-liquid chromatography, both sweet potato and soybean beta-amylase were found to convert maltal to the beta-anomer of 2-deoxymaltose. That maltal undergoes cis hydration provides evidence in support of a general-acid-catalyzed, carbonium ion mediated reaction. Of fundamental significance is that beta-amylase protonates maltal from a direction opposite that assumed for protonating starch, yet creates products of the same anomeric configuration from both. Such stereochemical dichotomy argues for the overriding role of protein structures in dictating the steric outcome of reactions catalyzed by a glycosylase, by limiting the approach and orientation of water or other acceptors to the reaction center.     

Hexameric purine nucleoside phosphorylase II from Escherichia coli K-12. Physico-chemical and catalytic properties and stabilization with substrates

Some properties of hexameric purine nucleoside phosphorylase II (EC 2.4.2.1) from Escherichia coli K-12 were studied. The enzyme obeys the Michaelis-Menten kinetics with respect to purine substrates (Km for inosine, deoxyinosine and hypoxanthine are equal to 492, 106 and 26.6 microM, respectively) and exhibits negative kinetic cooperativity towards phosphate and ribose-1-phosphate. The Hill coefficient is equal to approximately 0.5 for both substrates. Hexameric purine nucleoside phosphorylase II is not a metal-dependent enzyme; its activity is inhibited by Cu2+, Zn2+, Ni2+ and SO4(2-). The enzyme is the most stable at pH 6.0; it contains essential thiol groups. All substrates partly protect the enzyme against inactivation by 5.5'-dithiobis(2-nitrobenzoic acid) and heat-inactivation and, with the exception of phosphate-against inactivation by p-chloromercuribenzoate. Hypoxanthine, especially in combination with phosphate, afford the best protection against inactivation.     

Design of a growth model for yeast

The study of the cell cycle of a yeast strain made it possible to define two parameters:T, the time elapsing between the appearance of two consecutive buds on a mother cell, and Θ, the time elapsing between the appearance of a bud and the beginning of the first mitotic cycle. The influence of these two parameters on the growth rate of the strain is studied.