Structure of N-glycosidic carbohydrates of secretory proteins of Tetrahymena thermophila

Glycoproteins secreted by Tetrahymena into the culture medium were isolated and the N-glycosidic oligosaccharides analyzed using lectin blots and fluorophore-assisted carbohydrate gel electrophoresis (FACE). Lectin blots showed that the glycoproteins secreted by Tetrahymena contain only N-glycosidic structures of the high mannose type. Further analysis using the FACE technology revealed the presence of four different N-glycosidic structures differing only in the number of mannose residues attached to the core chitobiose unit.     

Evolution of C4 phosphoenolpyruvate carboxylase in Flaveria, a conserved serine residue in the carboxyl-terminal part of the enzyme is a major determinant for C4-specific characteristics

C4 phosphoenolpyruvate carboxylases have evolved from ancestral C3 isoforms during the evolution of angiosperms and gained distinct kinetic and regulatory properties compared with the C3 isozymes. To identify amino acid residues and/or domains responsible for these C4-specific properties the C4 phosphoenolpyruvate carboxylase of Flaveria trinervia (C4) was compared with its orthologue in the closely related C3 plant Flaveria pringlei. Reciprocal enzyme chimera were constructed and the kinetic constants, K(0.5) and k(cat), as well as the Hill coefficient, h, were determined for the substrate phosphoenolpyruvate both in the presence and absence of the activator glucose 6-phosphate. By this approach two regions were identified which determined most of the kinetic differences of the C4 and C3 ppcA phosphoenolpyruvate carboxylases with respect to the substrate PEP. In addition, the experiments suggest that the two regions do not act additively but interact with each other. The region between amino acids 296 and 437 is essential for activation by glucose 6-phosphate. The carboxyl-terminal segment between amino acids 645 and 966 contains a C4 conserved serine or a C3 invariant alanine at position 774 in the respective enzyme isoform. Site-directed mutagenesis shows that this position is a key determinant for the kinetic properties of the two isozymes.     

Influence of sodium butyrate on the induction of radiation-induced chromosomal aberrations and sister chromatid exchanges in Chinese hamster ovary (CHO) cells.

CHO cells were pre-treated with sodium butyrate (SB) for 24 h and then X-irradiated in G1. Metaphases were scored for the induction of chromosomal aberrations and sister chromatid exchanges (SCEs). The data were compared with those obtained after irradiation of cells not pre-treated with SB and showed that SB has different effects on the endpoints examined. The frequencies of dicentric chromosomes were elevated and of small acentric rings (double minutes, DMs) reduced. These results are discussed to be a consequence of conformational changes in hyperacetylated chromatin which could lead to more interchromosomal and to less intrachromosomal exchanges. SB itself induces a few SCEs but suppresses the induction of SCEs by X-rays. We assume that a minor part of radiation induced SCEs are 'false' resulting from structural chromosomal aberrations, such as inversions, induced in G1. Inversions are the symmetrical counterparts of DMs. If inversions are suppressed by SB treatment to a similar extent as DMs a small reduction of SCEs by SB can be expected.     

Studies on the progress of third-molar mineralisation in a Black African population.

The forensic determination of the age of living people has become increasingly important in recent years. With regard to the relevant age group, the radiographic assessment of third-molar mineralisation is of particular importance. So far, the influence of geographic origin on the mineralisation rate has been insufficiently analysed. The paper is based on a total of 595 conventional orthopantomograms of 474 male and 121 female Black Africans aged between 10 and 26 years for whom dates of birth were known. The mineralisation status of third molars was evaluated based on Demirjian's classification of stages [Demirjian et al., 1973. A new system of dental age assessment. Hum. Biol. 45, 221-227]. This study presents the means and standard deviations, median values and the lower and upper quartiles separately for both sexes for the mineralisation stages D-H. Statistically significant differences between the upper and lower jaws were observed in males examined with regard to their attaining the stage F. Mandibular teeth developed 0.8 years earlier than maxillary teeth. Significant sex-specific differences were found with regard to the age at which tooth 38 reached the stage G. In females, tooth 38 reached stage G 1.5 years earlier than in males. In comparison to White probands, the Black African sample showed a tendency to achieve the mineralisation stages earlier. We would recommend using population-specific standards for age determination purposes.     

Phytoplankton nitrogen demand and the significance of internal and external nitrogen sources in a large shallow lake (Lake Balaton,Hungary)

Since the middle of 1990s the trend of Lake Balaton towards an increasingly trophic status has been reversed, but N₂-fixing cyanobacteria are occasionally dominant, endangering water quality in summer. The sources of nitrogen and its uptake by growing phytoplankton were therefore studied. Experiments were carried out on samples collected from the middle of the Eastern (Siófok) and Western (Keszthely) basins between February and October 2001. Ammonium, urea and nitrate uptake and ammonium regeneration were measured in the upper 5-cm layer of sediment using the ¹⁵N-technique. Ammonium was determined by an improved microdiffusion assay. N₂ fixation rates were measured by the acetylene-reduction method. Ammonium regeneration rates in the sediment were similar in the two basins. They were relatively low in winter (0.13 and 0.16 μg N cm^?3 day^?1 in the Eastern and Western basin, respectively), increased slowly in the spring (0.38 and 0.45 μg N cm^?3 day^?1) and peaked in late summer (0.82 and 1.29 μg N cm^?3 day^?1, respectively). Ammonium uptake was predominant in spring in the Eastern basin and in summer in the Western basin, coincident with the cyanobacterial bloom. The amount of N₂ fixed was less than one third of the internal load during summer when external N loading was insignificant. Potentially, the phytoplankton N demand could be supported entirely by the internal N load via ammonium regeneration in the water column and sediment. However, the quantity of N from ammonium regeneration in the upper layer of sediment combined with that from the water column would limit the standing phytoplankton crop in spring in both basins and in late summer in the Western basin, especially when the algal biomass increases suddenly.     

Chromosome studies on lymphocytes of patients under cytostatic therapy

Summary Chromosome studies were performed on lymphocyte cultures from the peripheral blood of 43 patients undergoing a cytostatic interval therapy with a regimen of methyl-CCNU, 5-fluorouracil, and vincristine. From a total of 229 individual examinations (30 prior to the start of therapy, the rest at different stages of the therapy) more than 23 000 metaphases were analysed for structural chromosome aberrations (i.e., gaps, breaks, and interchanges).A distinct increase in the aberration rate over the level in controls (i.e., the same patients and other patients of the same group prior to the start of therapy) was observed at all phases of therapy in all patients under study. This increase was not only correlated with the number of therapy rounds, but was also dependent on the particular composition of the drug regimen applied at the different phases of therapy. Furthermore, even 4 weeks after 5-day courses of therapy the aberration rates, although lower than immediately after drug administration, clearly exceeded those of controls in most cases. There was no distinct quantitative difference, but there was a clear qualitative difference between data gained from 48-h cultures and those obtained after 72-h lymphocyte cultures. A rather high interindividual variability of aberration rates and spectra was established in different patients at the same phases of therapy.From these results the conclusion was drawn that chromosome studies in patients exposed to mutagenic chemicals are a suitable and reliable method of gaining insight into the clastogenicity of these substances in man (i.e., under the real conditions present in the human organism), if all the limitations of this method are seriously borne in mind.Abbreviations used methyl-CCNU 1-[2-chloroethyl-3-(4-methylcyclohexyl)]-1-nitrosourea - FU 5-fluorouracil - VC vincristine Dedicated to Prof. Dr. A. Barthelmeß on the occasion of his 70th birthday