Identity of the polymorphisms for esterase D and S-formylglutathione hydrolase in red blood cells

Summary The S-formylglutathione hydrolase (FGH) polymorphism of human red blood cells was studied in unrelated individuals, both by isoelectric focusing and starch gel electrophoresis, and with the substrates S-acetylglutathione and 4-methylumbelliferyl-acetate (the standard substrate for esterase D (ESD)). With both separation techniques the two substrates consistently gave similar and identically located zymograms. Thus, FGH (E.C.3.1.2.12) appears to be identical to ESD (E.C.3.1.1.1).     

Posttranslational modification of interleukin-2 is a late event during activation of human T lymphocytes by ionophore A23187 and phorbol ester

Human peripheral blood lymphocytes secrete high titers of interleukin-2 (IL-2) after stimulation by Ca2+-ionophore A23187/phorbol 12-myristate-13-acetate. During the first 30 hours of incubation cells secrete only the nonglycosylated IL-2 M form of the lymphokine, the glycosylated forms IL-2 N1,2 being detected only after prolonged culture times (30-48 h). After recultivation of cells for a second 48 h period (without additional mitogen), the glycosylated and nonglycosylated IL-2 forms are secreted at a constant ratio of 7:3 throughout. The detection of glycosylated IL-2 is parallelled by an increase in cellular glycosyltransferase activities involved in formation of sialylated oligosaccharides O-linked to proteins.     

Protein synthesis elongation factor 1 from rat liver: A zinc metalloenzyme

Highly purified elongation factor 1 (light form, EF1_L) from rat liver contains zinc as determined by atomic absorption spectrophotometry. Analysis has been performed on the most active protein fraction from DEAE-Sephadex chromatography (estimated purity: 90%) and on the main band obtained from this fraction by polyacrylamide gel electrophoresis. The data are consistent with a stoichiometry of approximately one g-atom of zinc per 54,000 daltons of EF1_L protein. A functional role for Zn²⁺ is suggested by the fact that 0.3 mM 1,10-phenanthroline completely abolishes GTP binding by EF1_L (measured by the nitrocellulose filter retention assay), while the isomeric non-chelator 1,7-phenanthroline has no effect. This inhibition can be overcome by the addition of excess zinc ion.     

Resolution of Conflicting Signals at the Single-Cell Level in the Regulation of Cyanobacterial Photosynthesis and Nitrogen Fixation

Unicellular, diazotrophic cyanobacteria temporally separate dinitrogen (N₂) fixation and photosynthesis to prevent inactivation of the nitrogenase by oxygen. This temporal segregation is regulated by a circadian clock with oscillating activities of N₂ fixation in the dark and photosynthesis in the light. On the population level, this separation is not always complete, since the two processes can overlap during transitions from dark to light. How do single cells avoid inactivation of nitrogenase during these periods? One possibility is that phenotypic heterogeneity in populations leads to segregation of the two processes. Here, we measured N₂ fixation and photosynthesis of individual cells using nanometer-scale secondary ion mass spectrometry (nanoSIMS) to assess both processes in a culture of the unicellular, diazotrophic cyanobacterium Crocosphaera watsonii during a dark-light and a continuous light phase. We compared single-cell rates with bulk rates and gene expression profiles. During the regular dark and light phases, C. watsonii exhibited the temporal segregation of N₂ fixation and photosynthesis commonly observed. However, N₂ fixation and photosynthesis were concurrently measurable at the population level during the subjective dark phase in which cells were kept in the light rather than returned to the expected dark phase. At the single-cell level, though, cells discriminated against either one of the two processes. Cells that showed high levels of photosynthesis had low nitrogen fixing activities, and vice versa. These results suggest that, under ambiguous environmental signals, single cells discriminate against either photosynthesis or nitrogen fixation, and thereby might reduce costs associated with running incompatible processes in the same cell.     

Minimal algal food requirements in the presence of protozoan prey for the rotifer Brachionus calyciflorus

As recent experiments demonstrated that protozoans support reproductionof the rotifer Brachionus calyciflorus only in the presenceof algal prey, we addressed the question of minimal algal foodrequirements in a mixed diet of algae and protozoans to ensurepositive growth of B. calyciflorus. In numerical response experiments,we determined the algal threshold concentration for zero populationgrowth of B. calyciflorus when feeding on a pure algal dietof Monoraphidium minutum, and on a mixed diet of M. minutumplus the ciliate Coleps sp. Under pure algal and under mixedfood conditions, a minimum amount of 0.3 µg carbon (C)ml^-1 M. minutum was needed to ensure zero population growthof B. calyciflorus. At lower algal concentrations, Coleps sp.was of low nutritional value. However, when offered in conjunctionwith a concentration of 0.8 µg C ml^-1 M. minutum, Colepssp. contributed equally to the reproductive success of B. calyciflorusas compared to a sole M. minutum diet. Overall, Coleps sp. canbe an adequate supplement in the diet of B. calyciflorus ifa sufficient amount of algal prey is available. However, attimes of low phytoplankton biomass, consumption of ciliatesdoes not necessarily confer a reproductive advantage for thisrotifer.     

Adrenomedullin is a novel marker of tumor progression in neuroendocrine carcinomas.

Adrenomedullin is a multi-functional polypeptide hormone. Its involvement in angiogenesis and vasodilator action support the hypothesis that adrenomedullin may be a secretory product of neuroendocrine tumors and contribute to tumor progression. Plasma levels of adrenomedullin were measured by radioimmunoassay in 46 patients with neuroendocrine carcinomas of the gastroenteropancreatic and bronchial system. Tissue expression of adrenomedullin was studied using monoclonal antibodies on pretreated paraffin embedded tissues in a group of 31 patients. Adrenomedullin plasma levels were significantly elevated in patients compared to healthy age-matched controls (p < 0.001). The highest plasma levels were found in patients with neuroendocrine carcinomas of bronchial, midgut and unknown origin. Patients with progressive disease had higher plasma levels than patients with stable disease (p < 0.001). Of the examined tumor samples, 55 % showed cytoplasmic staining for adrenomedullin > 5 % of the total tumor area. Plasma levels and tissue expression of adrenomedullin did not correlate with functional activity of the tumors or presence of the carcinoid syndrome, but did with tumor progression (p < 0.001 and p < 0.014). In conclusion, plasma and tissue expression of the angiogenic peptide adrenomedullin are predictive of tumor progression in patients with neuroendocrine carcinomas. Adrenomedullin might represent a useful prognostic marker in patients with neuroendocrine carcinomas.