A cyclic 3',5'-adenosine monophosphate-binding protein in the causative agent of plague

cAMP-binding protein was isolated from the plaque agent and purified to the homogeneous state. Purification process included filtration of the initial preparation through the membrane able to transmit particles with mol. weight to 300,000 Da, chromatography on cellulose "DE-52" and biogel HTP. The protein homogeneity was confirmed by electrophoresis in polyacrylamide gel and precipitation with commercial plague agglutinating serum. The protein with mol. weight of 180,000 Da consisted of two identical subunits (90,000 Da. each) which could dissociate with formation of monomers (mol. weight approximately 18,000 Da), Cu2+, Co2+, Mn2+ ions stimulated activity of cAMP-binding protein of a plague microbe while Fe3+, Ca2+, Zn2+ ions inhibited it by 30-70%. A monospecific rabbit serum to the homogeneous preparation of cAMP-binding protein was obtained. It helped finding the similar protein in the close relative bacterium Yersinia pseudotuberculosis but not in Y. enterocolitica.     

Elemental distribution in striated muscle and the effects of hypertonicity: Electron probe analysis of cryo sections

A method of rapid freezing in supercooled Freon 22 (monochlorodifluoromethane) followed by cryoultramicrotomy is described and shown to yield ultrathin sections in which both the cellular ultrastructure and the distribution of diffusible ions across the cell membrane are preserved and intracellular compartmentalization of diffusabler ions can be quantitated. Quantitative electron probe analysis (Shuman, H., A.V. Somlyo, and A.P. Somlyo. 1976. Ultramicros. 1:317-339.) of freeze-dried ultrathin cryto sections was found to provide a valid measure of the composition of cells and cellular organelles and was used to determine the ionic composition of the in situ terminal cisternae of the sarcoplasmic reticulum (SR), the distribution of CI in skeletal muscle, and the effects of hypertonic solutions on the subcellular composition if striated muscle. There was no evidence of sequestered CI in the terminal cisternae of resting muscles, although calcium (66mmol/kg dry wt +/- 4.6 SE) was detected. The values of [C1](i) determined with small (50-100 nm) diameter probes over cytoplasm excluding organelles over nuclei or terminal cisternae were not significantly different. Mitochondria partially excluded C1, with a cytoplasmic/ mitochondrial Ci ratio of 2.4 +/- 0.88 SD. The elemental concentrations (mmol/kg dry wt +/- SD) of muscle fibers measured with 0.5-9-μm diameter electron probes in normal frog striated muscle were: P, 302 +/- 4.3; S, 189 +/- 2.9;C1, 24 +/- 1.1;K, 404 +/- 4.3, and Mg, 39 +/- 2.1. It is concluded that: (a) in normal muscle the "excess CI" measured with previous bulk chemical analyses and flux studies is not compartmentalized in the SR or in other cellular organelles, and (b) the cytoplasmic C1 in low [K](0) solutions exceeds that predicted by a passive electrochemical distribution. Hypertonic 2.2 X NaCl, 2.5 X sucrose, or 2.2 X Na isethionate produced: (a) swollen vacuoles, frequently paired, adjacent to the Z lines and containing significantly higher than cytoplasmic concentrations of Na and Cl or S (isethionate), but no detectable Ca, and (b) granules of Ca, Mg, and P = approximately (6 Ca + 1 Mg)/6P in the longitudinal SR. It is concluded that hypertonicity produces compartmentalized domains of extracellular solutes within the muscle fibers and translocates Ca into the longitudinal tubules.     

Effect of HCO3- and carbon dioxide at various concentrations on activity of certain enzymes

The paper deals with the effect of changes in the concentration of carbonic acid in the medium on the reaction rate catalyzed with enzymes of various spectrum of the action. It is shown that the presence of carbonic acid in the medium reaction increases the rate of reactions catalyzed with lactate dehydrogenase of the rabbit liver soluble fraction, with glucose-6-phosphate dehydrogenase from yeast and trypsin. Under the same conditions the reaction rate catalyzed with glucose-6-phosphate dehydrogenase of the rabbit liver soluble fraction and with ATP-citrate (pro-3S)-lyase is considerably decreased. Changes in the carbonic acid concentrations within the physiological limits are found to have no effect on lactate dehydrogenase from the cattle heart and chymotrypsin.     

Association connections of the cat's parietal cortex

Intercortical connections of primary sensory (visual, auditory, somatosensory) areas with the parietal association cortex were studied in cats by the retrograde axonal transport of horseradish peroxidase and the Fink-Heimer silver impregnation of degenerated fibers techniques. This combined study revealed the shape, size, and intracortical location of cells connecting the primary sensory areas monosynaptically with the parietal cortex and also the distribution of preterminals and terminals of the fibers of these cells in the parietal association cortex. The greatest number of cells forming connections with area 7 of the parietal association cortex was shown to occur in visual area V1, and with area 5 in somatosensory area S1. Besides pyramidal neurons tagged with horseradish peroxidase, which were located mainly in layers II–IV, a few tagged stellate and fusiform cells also were found. The results supplement and confirm data on afferent connections of the parietal association cortex in cats.M. Gor'kii Donetsk Medical Institute. Translated from Neirofiziologiya, Vol. 13, No. 1, pp. 3–6, January, 1981.     

Role of immune cytolysis and its thymosin stimulation in experimental influenza

In experiments on C57BL and CC57W mice the acute or chronic course of experimental influenza infection has been shown to correlate with the activity of immune cytolysis. At a low level of the cytolytic activity of T-lymphocytes the prolonged persistence of influenza virus develops. The stimulation of cell-mediated immunity with thymosin prevents the development of chronic influenza infection.     

Sequential changes in the macrostructure and RNA-synthesizing activity of nucleolar and extranucleolar chromatin from rat liver cells during induction of DNA synthesis

The dynamics of structural changes and RNA-polymerase activity in rat liver cell chromatin caused by drastic changes in the rates of protein synthesis was investigated. Inhibition of protein synthesis after a single injection of animals with cycloheximide (0.3 mg/100 g of body weight) increased the total condensibility of chromatin. Under these conditions, the stepwise activation of RNA-polymerases I and II correlated with decondensation of chromatin. By the 6-12th hour following cycloheximide injection, a chromatin fraction enriched with RNA-polymerase I and a RNA-polymerase II-rich fraction could be isolated from liver cell nuclei.