Generation of a transmembrane gradient of Na+ in Methanosarcina barkeri

A transmembrane Na+ gradient was generated by Methanosarcina barkeri during methanogenesis. The intracellular Na+ concentration amounted to approximately one fifth of the extracellular one. A secondary Na+/H+ antiport system was shown to be responsible for Na+ extrusion. This system could be inhibited by amiloride. In the presence of amiloride the delta pH across the cytoplasmic membrane increased and a transmembrane Na+ gradient could neither be generated nor maintained. The possible role of Na+ in the oxidation of methanol to the level of formaldehyde is discussed.     

Ethanol induces release of arachidonic acid but not synthesis of eicosanoids in mouse peritoneal macrophages

Exposure of mouse peritoneal macrophages to ethanol induces a rapid release of arachidonic acid to the extracellular medium. All major classes of phospholipids, phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol contribute to this release. Ethanol-induced mobilization of arachidonic acid occurs by deacylation, but it is not accompanied by eicosanoid synthesis. These data suggest that at least two signals are necessary for the release and metabolism of arachidonic acid. Ethanol also activates a phospholipase C which hydrolyzes only phosphatidylinositol, and not its phosphorylated derivatives.     

Influence of Ca2+ and Mg2+ on the turnover of the phosphomonoester group of phosphatidylinositol 4-phosphate in human erythrocyte membranes.

In isolated erythrocyte membranes, increasing the free Mg2+ concentration from 0.5 to 10 mM progressively activates the membrane-bound phosphatidylinositol (PtdIns) kinase and leads to the establishment of a new equilibrium with higher phosphatidylinositol 4-phosphate (PtdIns4P) and lower PtdIns concentrations. The steady-state turnover of the phosphomonoester group of PtdIns4P also increases at high Mg2+ concentrations, indicating a simultaneous activation of PtdIns4P phosphomonoesterase by Mg2+. Half-maximum inhibition of PtdIns kinase occurs at 10 microM free Ca2+ in the presence of physiological free Mg2+ concentrations. Increasing free Mg2+ concentrations overcome Ca2+ inhibition of PtdIns kinase. In the presence of Ca2+, calmodulin activates Ca2+-transporting ATPase 5-fold, but does not alter pool size and radiolabelling of PtdIns4P. In intact erythrocytes, adding EGTA or EGTA plus Mg2+ and the ionophore A23187 to the external medium does not exert significant effects on concentration and radiolabelling of polyphosphoinositides when compared with controls in the presence of 1.4 mM free Ca2+.     

Radioactive labeling techniques for the identification of G antigen in the human Rh blood group system

The G antigen is one of the erythrocyte membrane Rh antigens. The amount of Rh antigen present on the red blood cell is about 10(-15) g and radioactive labeling of membrane proteins is a useful method for its identification and characterization. In this paper, we compare 4 labeling techniques. Using a human monoclonal anti-Rh(G) antibody and an immunofixation technique, we located the G antigen on a polypeptide of an average molecular weight of 28,000 Da.     

Three-dimensional representation of chemical gradients

Perspective display techniques are applied to chemical and biochemical data sets. These represent spatially distributed gradients of reactive compounds that participate in pattern-formation processes due to reaction-diffusion or reaction-convection coupling. The patterns form in thin solution layers and are observed as chemical waves in the Belousov-Zhabotinskii reaction, as convection-induced stationary structures during oscillating glycolysis in yeast cytoplasm, and as the diffusive spreading of enzyme-catalyzed metabolic turnover in a substrate layer. The digital data are measured with a two-dimensional spectrophotometer based on a computerized video equipment with high spatial, temporal and intensity resolution. By application of three-dimensional procedures detailed structural properties of chemical and biochemical model systems will be presented yielding localization of reaction and transport events.     

Osmoregulation in Escherichia coli by accumulation of organic osmolytes: betaines,glutamic acid,and trehalose

Like other cyanobacteria, Chlorogloeopsis fritschii contained as major lipid classes monogalactosyldiacyl-glycerols, digalactosyldiacylglycerols, sulfoquinovosyldiacylglycerols and diacylglycerophosphoglycerols. In addition to these lipid classes this cyanobacterium also contained small amounts of diacylglycerophosphocholines and sterols, predominantly lanosterol, thus showing similarity to photosynthetic eukaryotes. Dark incubated cells contained larger proportions of the latter two lipid classes than light grown cells. Like other prokaryotes, C. fritschii lacked linolenic acid (18:3) in its lipids. Lipids from the thylakoids were richer in palmitoleic acid (16:1) than those of whole cells. There was no effect of light on the patterns of constituent fatty acids of lipids from C. fritschii, in contrast to photosynthetic eukaryotes.Abbreviations MGDG Monogalactosyldiacylglycerols - PA Phosphatidic acids - PE Diacylglycerophosphoethanolamines - PG Diacylglycerophosphoethanolamines - DGDG Digalactosyldiacylglycerols - SQDG Sulfoquinovosyldiacylglycerols - PC Diacylglycerophosphocholines     

Three new orosomucoid (ORM) variants revealed by isoelectric focusing and print immunofixation

Summary Phenotypes of orosomucoid (ORM) in human sera have been analysed by isoelectric focusing and print immunofixation. After neuraminidase treatment the band patterns indicated that the polymorphism of the structural locus ORM1 is controlled by three autosomal codominant alleles. According to the previous nomenclature they were called ORM1^*F1, ORM1^*F2, and ORM1^*S. In a study of 272 unrelated individuals from southern Germany, five of the six expected common ORM1 subtypes were observed. Furthermore, we found three ORM variant phenotypes which have not been reported previously. These variants were characterized by additional bands in a cathodal position. One variant had additional double bands and presumably represents a rare ORM1 variant named ORM1S1. Two variants had additional single bands. They were assigned tentatively to the ORM2 gene locus. While the common gene product of ORM2 may be called ORM2A, the two variants are named ORM2B1 and ORM2B2, respectively. ORM2B1 has, thus far, been found only in a single individual; the variants ORM1S1 and ORM2B2 were found in a father-child pair and a mother-child pair, respectively. The frequency for variants tentatively assigned to the ORM2 locus is very low and was calculated to be 0.0037.     

Structural analysis of the phytophagous insect guilds associated with the roots of Centaurea maculosa Lam. C. diffusa Lam., and C. vallesiaca Jordan in Europe:

Summary During extensive field surveys in central and eastern Europe, 21 herbivorous root insect species were found on Centaurea maculosa ssp. rhenana Boreau, 12 species on C. diffusa Lam. and 11 species on C. vallesiaca Jordan, representing 12 families in 4 orders. The large geographic distribution (species-area function), the high number of Centaurea spp. present (host speciation rate), and the high apparency of the rosettes and the rich food resources offered by the roots during winter, together with their poor accessibility, correlate with the high number of specialist feeders associated with the roots of C. maculosa and C. diffusa. The members of the taxonomically diverse root entomofauna exploit specific structures of the tap root (food niches). Interspecific competition among members of food niches, as well as species-specific responses to different phenological stages (for oviposition) and tissues (for larval development) are thought to be responsible for the high predictability in guild structure. The relatively low levels of host plant attack (two thirds of the roots were unattacked) and the fact that food niches remained unoccupied in most of the regions suggest, however, that the majority of the studied guilds do not represent equilibrium assemblages. Ecological (different habitats), climatic (transitional zone) and historical (ancient pre-Pleistocene communities) factors could account for the highest values of species diversity, infestation levels, species packing and food niche utilization, which are found on C. maculosa in E. Austria/NW. Hungary, compared to other regions. A positive correlation between species packing (number of root-feeding species per population) and infestation rates (percent of roots attacked) was only found for the more stable, semi-natural habitats. A comparative analysis of the regional root insect guilds of C. maculosa with corresponding data for the phytophagous insects associated with the flower heads revealed distinct taxonomical differences, but a high degree of numerical and structural similarity. The different geographical regions are similarly ranked for host plant attack, herbivore pressure, average species packing and level of food niche utilization.     

Sucrose transport into the phloem of Ricinus communis L. seedlings as measured by the analysis of sieve-tube sap

Careful cutting of the hypocotyl of Ricinus communis L. seedlings led to the exudation of pure sieve-tube sap for 2–3 h. This offered the possibility of testing the phloem-loading system qualitatively and quantitatively by incubating the cotyledons with different solutes of various concentrations to determine whether or not these solutes were loaded into the sieve tubes. The concentration which was achieved by loading and the time course could also be documented. This study concentrated on the loading of sucrose because it is the major naturally translocated sieve-tube compound. The sucrose concentration of sieve-tube sap was approx. 300 mM when the cotyledons were buried in the endosperm. When the cotyledons were excised from the endosperm and incubated in buffer, the sucrose concentration decreased gradually to 80–100 mM. This sucrose level was maintained for several hours by starch breakdown. Incubation of the excised cotyledons in sucrose caused the sucrose concentration in the sieve tubes to rise from 80 to 400 mM, depending on the sucrose concentration in the medium. Thus the sucrose concentration in the sieve tubes could be manipulated over a wide range. The transfer of labelled sucrose to the sieve-tube sap took 10 min; full isotope equilibration was finally reached after 2 h. An increase of K⁺ in the medium or in the sieve tubes did not change the sucrose concentration in the sievetube sap. Similarly the experimentally induced change of sucrose concentration in the sieve tubes did not affect the K⁺ concentration in the exudate. High concentrations of K⁺, however, strongly reduced the flow rate of exudation. Similar results were obtained with Na⁺ (data not shown). The minimum translocation speed in the sieve tubes in vivo was calculated from the growth increment of the seedling to be 1.03 m·h^-1, a value, which on average was also obtained for the exudation system with the endosperm attached. This comparison of the in-vivo rate of phloem transport and the exudation rate from cut hypocotyls indicates that sink control of phloem transport in the seedlings of that particular age was small, if there was any at all, and that the results from the experimental exudation system were probably not falsified by removal of the sink tissues.Abbreviations PTS 3-hydroxy-5,8, 10-pyrenetrisulfonate