The distribution and interconversion of ammonium and nitrate in the Skallingen salt marsh (Denmark) and their exchange with the adjacent coastal water

The potential nitrogen sources for the primary production in the intertidal area are nitrogen compounds obtained from mineralization in the sediment and the water column, nitrogen fixation, outflow from rivers and groundwater seeping from the mainland. The available inorganic nitrogen in the adjacent coastal waters decreases from 50–80 μmol NO₃ ^-/l and 6–15 μmol NH₄ ⁺/l in early spring to ca one tenth during the growing season. In the sediment of the tidal flats available ammonia and nitrate vary between 50 and 100 μmol/1 pw. In the salt marsh available ammonia increases from 200–300 nmol NH₄ ⁺/g fwt to approximately double the amount, and the available nitrate varies from 100–300 nmol NO₃ ^-/g fwt (250–750 μmol NO₃ ^-/l pw) to ca one third during the growing season. The exchange of NH₄ ⁺, NO₂ ^- and NO₃ ^- across the sediment water interface has been estimated during tidal cycles under light and dark conditions on the tidal flats. The flux of nitrogen was dependent on the flora and fauna as well as the time of the year. The tidal activity, frequency and length of inundation are considered the driving force in a two-way process between salt marshes and adjacent coastal waters. The role of marsh sediment, tidal water and sediments of the tidal flats as sites of accumulation, consumption and remineralization of organic matter is emphasized. The possible exchange of ammonia and nitrate between the salt marsh and the different compartments of the tidal water is discussed.     

白头翁的受精及组织化学研究

成熟花粉为二细胞,生殖细胞的蛋白质染色较营养细胞的深,淀粉粒充满营养细胞。花粉管常见穿人正在退化中的助细胞,并释放出两个精子,大量稠浓的蛋白质和淀粉,个别的释放在助细胞与胚囊壁之间。在一些卵细胞核,次生核(或极核)中,受精前后有1—3个小核仁(约1微米);助细胞具丝状器;反足细胞宿存,具多核和多核仁。胚囊中各个细胞的原生质稠浓程度和蛋白质染色深浅不同,其中以反足细胞和助细胞的原生质最稠浓,蛋白质染色最深。淀粉十分贫乏,绝大多数卵细胞,中央细胞和几乎全部助细胞和反足细胞均不含淀粉。双受精属于有丝分裂前类型,两性核融合步骤为:精子核接近和贴附在卵核和次生核上(或极核上),两性核的核膜溶解,其染色质沉入融合核,并随之松解,同时出现雄性核仁,最终,雄性核仁和雌性核仁合并,形成具单核和单核仁的合子和初生胚乳细胞。另外,雄性核仁同卵核仁合并较雄性核仁和次生核的晚,所以卵细胞完成受精较次生核晚。     

The effect of temperature on the acetylcholinesterase activity of muscle microsomes

Membrane vesicles which constitute the sarcotubular system were separated and the fraction enriched in T-tubules purified by a calcium loading procedure. The preparations of unfractioned microsomes and T-tubules have been analyzed for their relative content of enzyme markers and acetylcholinesterase. The amount of this enzyme in the T-tubule fraction was higher than in mixed microsomes but less than two-fold the value of vesicles derived from sarcoplasmic reticulum. Arrhenius plots of membrane-bound and soluble acetylcholinesterase from either mixed microsomes or fractions enriched in T-tubules show an anomalous behaviour as two break points were obtained. The first discontinuity was found at about 17 degrees C for membrane-bound, and 12-14 degrees C for soluble acetylcholinesterase. The second one being at about 25 degrees C for both particulate and detergent-solubilized enzyme. The changes in activity with temperature suggest that lipid-protein, detergent-protein and protein-protein interactions might be involved in the stabilization of the enzyme both in the natural membrane and in the soluble state.     

Extractive bioconversion in aqueous two-phase systems

Summary The transformation of hydrocortisone to prednisolone was studied in aqueous two-phase systems, as a model for the extractive bioconversion of fine chemicals. The bacterium, Arthrobacter simplex, was able to grow in the two-phase system and the cells could be revitalized after a period of use. Use of aqueous two-phase systems made it possible to operate the reaction at higher substrate concentrations than in pure buffer solution. An adsorptive method to remove the product from the top phase was tested and shown to be both efficient and compatible with the overall process. In order to reduce the costs of operation in aqueous two-phase systems, a cheaper starch-based polymer, Reppal-PES, was successfully used as a substitute for dextran.Dedicated to Professor Dr. Georg Manecke on the occasion of his 70th birthday     

Study of the hydrolysis and ionization constants of Schiff base from pyridoxal 5''-phosphate and n-hexylamine in partially aqueous solvents. An application to phosphorylase b.

Formation and hydrolysis rate constants as well as equilibrium constants of the Schiff base derived from pyridoxal 5'-phosphate and n-hexylamine were determined between pH 3.5 and 7.5 in ethanol/water mixtures (3:17, v/v, and 49:1, v/v). The results indicate that solvent polarity scarcely alters the values of these constants but that they are dependent on the pH. Spectrophotometric titration of this Schiff base was also carried out. We found that a pKa value of 6.1, attributed in high-polarity media to protonation of the pyridine nitrogen atom, is independent of solvent polarity, whereas the pKa of the monoprotonated form of the imine falls from 12.5 in ethanol/water (3:17) to 11.3 in ethanol/water (49:1). Fitting of the experimental results for the hydrolysis to a theoretical model indicates the existence of a group with a pKa value of 6.1 that is crucial in the variation of kinetic constant of hydrolysis with pH. Studies of the reactivity of the coenzyme (pyridoxal 5'-phosphate) of glycogen phosphorylase b with hydroxylamine show that this reaction only occurs when the pH value of solution is below 6.5 and the hydrolysis of imine bond has started. We propose that the decrease in activity of phosphorylase b when the pH value is less than 6.2 must be caused by the cleavage of enzyme-coenzyme binding and that this may be related with protonation of the pyridine nitrogen atom of pyridoxal 5'-phosphate.     

The carbohydrate of human thrombin: Structural analysis of glycoprotein oligosaccharides by mass spectrometry

The carbohydrate structure of human thrombin has been determined by direct probe mass spectrometry of the oligosaccharides released by trifluoroacetolysis from the asialo glycoprotein. The free oligosaccharides were studied as permethylated and N-trifluoroacetylated oligosaccharide alditols. The structure was confirmed by sequential exoglycosidase digestion of intact thrombin and sugar and methylation analysis of the oligosaccharides by gas-liquid chromatography-mass spectrometry. The results indicate the following structure:

with Fuc present on only about 50% of the oligosaccharides.     

Effect of interferon treatment on blockade of protein synthesis induced by poliovirus infection

Treatment of HeLa cells with lymphoblastoid interferon leads to a drastic inhibition of infective poliovirus. Even relatively high concentrations of human lymphoblastoid interferon HuIFN-alpha (Ly) (400 IU/ml) do not prevent destruction of the cell monolayer after most of the cells have been infected with poliovirus. Analysis of macromolecular synthesis in a single step growth cycle of poliovirus in interferon-treated cells detected no viral protein synthesis. In spite of this inhibition of viral translation, the shut-off of host protein synthesis in interferon-treated cells is apparent when they are infected both at low and high multiplicities. Although viral RNA synthesis is inhibited considerably in cells treated with interferon, a certain amount is detected, suggesting that some viral replication takes place. Analysis of membrane permeability after poliovirus infection shows a leakage to 86Rb+ ions and modification of membrane permeability to the translation inhibitor hygromycin B at the moment when the bulk of virus protein synthesis occurs. These changes are delayed and even prevented if cells are pretreated with interferon. A situation is described in which host protein synthesis is shut-down with no major changes in membrane permeability, as studied by the two tests mentioned above. Prevention of viral gene expression by inactivation with ultraviolet light of the input virus or by treatment with cycloheximide blocks the shut-off of protein synthesis. This does not occur in the presence of 3 mM guanidine. These observations are in agreement with the idea that some poliovirus protein synthesis takes place in interferon-treated cells and this early gene expression is necessary to block cellular protein synthesis.     

Colorless Sulfur Bacteria, Beggiatoa spp. and Thiovulum spp., in O2 and H2S Microgradients

The interactions between colorless sulfur bacteria and the chemical microgradients at the oxygen-sulfide interface were studied in Beggiatoa mats from marine sediments and in Thiovulum veils developing above the sediments. The gradients of O₂, H₂S, and pH were measured by microelectrodes at depth increments of 50 μm. An unstirred boundary layer in the water surrounding the mats and veils prevented microturbulent or convective mixing of O₂ and H₂S. The two substrates reached the bacteria only by molecular diffusion through the boundary layer. The bacteria lived as microaerophiles or anaerobes even under stirred, oxic water. Oxygen and sulfide zones overlapped by 50 μm in the bacterial layers. Both compounds had concentrations in the range of 0 to 10 μmol liter⁻¹ and residence times of 0.1 to 0.6 s in the overlapping zone. The sulfide oxidation was purely biological. Diffusion calculations showed that formation of mats on solid substrates or of veils in the water represented optimal strategies for the bacteria to achieve a stable microenvironment, a high substrate supply, and an efficient competition with chemical sulfide oxidation. The continuous gliding movement of Beggiatoa cells in mats or the flickering motion of Thiovulum cells in veils were important for the availability of both O₂ and H₂S for the individual bacteria.