Long-term effects of postovulatory aging of mouse oocytes on offspring: a two-generational study.

Aims of this study were to analyze the long-term effects of postovulatory aging of mouse oocytes on 1) reproductive traits of parental (F(0)) and first (F(1))-generation females (pregnancy rate, gestation length, litter size, perinatal death, and sex ratio of offspring) and 2) developmental and behavioral variables of F(1) and second-generation (F(2)) offspring (birth weight and weight gain during preweaning development, postnatal day of attainment of immediate righting, spontaneous motor activity, and passive and active conditioned learning ability). Hybrid (C57BL/6JIco x CBA/JIco) females were artificially inseminated at 13 h (control group) or 22 h (oocyte-aged group) after GnRH injection. Experimental (oocyte-aged group) F(0) females exhibited lower pregnancy rate, shortened gestation length, decreased litter size, higher perinatal death of their pups, and increased percentage of male offspring compared to control F(0) females. Postovulatory aging of oocytes was also associated with increased number of growth-retarded pups, delayed development of the righting reflex, and higher spontaneous motor activity and emotionality of F(1) offspring. Postovulatory aging of F(0) oocytes did not affect birth weight, weight gain during preweaning development, passive and active conditioned learning ability of F(1) offspring, or reproductive traits of F(1) females or developmental and behavior variables of F(2) offspring.     

A new coumarin from Ferula loscosii and the correct structure of coladonin

From the roots of Ferula loscosii the coumarins umbelliprenin, coladin and coladonin and the new natural isovaleryl derivative of the latter have been isolated. A study which confirms the structure of coladonin and its difference from farnesiferol A is also given.     

Ultrastructural studies of endocrine-like cells in the fundic gastric mucosa of the bullfrog,Rana catesbeiana

Summary This study is concerned with electron-microscopic observations on endocrine or paracrine cells in the fundic gastric mucosa of the bullfrog. Also, an attempt was made to identify the histamine-releasing cells involved in the secretagogue response. At least three distinct endocrine-like cell types were found. The classification is based on the appearance of secretory granules and other organelles, and the relationship of endocrine-like cells with other cells in the tissue. The amphibian endocrine-like cells resemble the ECL, D and EC cells of mammals. Type-I (ECL) cells showed degranulation after repeated stimulation with tetragastrin (TG), acetylcholine (ACh) and K⁺ depolarizing solution, all of which release histamine.     

Production of vanillic acid from vanillin by resting cells of Serratia marcescens.

Resting-cell suspensions of Serratia marcescens were able to convert, quantitatively, 0.3% vanillin to vanillic acid. The vanillic acid-producing activity reached a maximum after 28 h of incubation with 0.01% vanillin as an inducer.     

Effect of acute altitude hypoxia on fatty acid proportion in the plasma of rats of different ages

Using gas chromatography, the authors studied the development of the proportion of 25 saturated and non-saturated fatty acids in the plasma of Wistar rats of their own breed aged 5, 10, 14 and 25 days and adult (both sexes), and in three age groups (5 and 14 day and adult) they determined the effect of acute altitude hypoxia. Fatty acids were determined as methylesters (FAME) (Base 1978), using the method already described by Smídová et al. (1984a), in mixed blood plasma obtained from decapitated control and experimental animals. Altitude hypoxia, which corresponded to 9 000 m, lasted 20 min. The temperature was kept stable at 25 degrees C. Marked changes occurred during ontogenesis in the proportion of total fatty acids (TFA) in the plasma; the proportion of sigma non-saturated FA fell significantly during maturation, chiefly in favour of monoene FA, with a resultant significant drop in their reciprocal index (sigma saturated/sigma monoene). The proportion of all saturated FA amounted in 5-day-old rats to 54% of the total quantity, but in adult rats to only 40%. Acute hypoxia had a demonstrable effect in all three age categories. In 5-day-old rats it led to significant changes in 7 FA and in adult rats in 10. In both 5-day-old and adult animals it produced an increase in the arachidonic acid level. The results are discussed in relation to previous findings on the effect of the same hypoxia on FA proportion in various parts of the CNS of rats of different ages.     

The formation of L-3-phosphoglyceric acid by ribulose-1,5-bisphosphate carboxylase

A sensitive and reliable method to determine the stereochemical composition of 3-phosphoglyceric acid is presented. Results obtained with this method show that 3-phosphoglyceric acid formed in the ribulose-1,5-bisphosphate carboxylase reaction is a mixture of 10% L-3-PGA and 90% D-3-PGA.     

Role of Asp-9 and Glu-36 in the active site of the pneumococcal CPL1 lysozyme: an evolutionary perspective of lysozyme mechanism.

The role of carboxylic amino acids Asp-9 and Glu-36 in the activity of CPL1 lysozyme was investigated by site-directed mutagenesis. The enzymatic activity of the single mutants D9E, D9N, D9H, D9K, D9A, E36D, E36Q, E36K, and E36A and of the double mutant D9A-E36A was analyzed using a highly sensitive radioactive assay. All mutants but D6K showed detectable activities. Interestingly, the mutants E36D and E36Q retained 67% and 37% activity, respectively. Amino acid replacements at position 9 turned out to be more critical for activity than at position 36. In analogy to the mechanism described for hen egg-white lysozyme, where the proton donor play a central role, we propose that, in the CPL1 lysozyme, Asp-9 might act as the proton donor for activation of the substrate, and Glu-36 could help in the stabilization of the intermediate oxocarbocation. The residual activity of lysozyme mutants lacking one or two of the acidic amino acids may be explained by the participation of a water molecule as proton donor and/or to electrostatic contributions in the active center stabilizing the transition state of the reaction. Our results are in agreement with the hypothesis that enzymes have been optimized during evolution from an ancestral protein able to bind more tightly the transition state of the substrate than the substrate itself, by the acquisition of amino acids serving a function in catalysis.