Galactocerebroside is expressed by non-myelin-forming Schwann cells in situ

Interest in the glycosphingolipid galactocerebroside (GC) is based on the consensus that in the nervous system it is expressed only by myelin-forming Schwann cells and oligodendrocytes, and that it has a specific role in the elaboration of myelin sheaths. We have investigated GC distribution in two rat nerves--the sciatic, containing a mixture of myelinated and non-myelinated axons, and the cervical sympathetic trunk, in which greater than 99% of axons are non-myelinated. Immunohistochemical experiments using mono- and polyclonal GC antibodies were carried out on teased nerves and cultured Schwann cells, and GC synthesis was assayed biochemically. Unexpectedly, we found that mature non-myelin-forming Schwann cells in situ and in short-term cultures express unambiguous GC immunoreactivity, comparable in intensity to that of myelinated fibers or myelin-forming cells in short-term cultures. GC synthesis was also detected in both sympathetic trunks and sciatic nerves. In the developing sympathetic trunk, GC was first seen at day 19 in utero, the number of GC-positive cells rising to approximately 95% at postnatal day 10. In contrast, the time course of GC appearance in the sciatic nerve shows two separate phases of increase, between day 18 in utero and postnatal day 1, and between postnatal days 20 and 35, at which stage approximately 94% of the cells express GC. These time courses suggest that Schwann cells, irrespective of subsequent differentiation pathway, start expressing GC at about the same time as cell division stops. We suggest that GC is a ubiquitous component of mature Schwann cell membranes in situ. Therefore, the role of GC needs to be reevaluated, since its function is clearly not restricted to events involved in myelination.     

Factors influencing respiration data in freshwater sediment

Sediment respiration (oxygen consumption and CO₂ evolution) was measured in freshwater sediment samples using both flask- and core-microcosms, and the estimates were compared. Oxygen consumption data were also compared in flask-microcosms constructed with sediment samples of different masses, sediment: water ratios, and storage times. Furthermore, sediment respiration was examined under different incubation conditions of temperature and agitation. O₂ consumption was markedly higher in flask-microcosms than in sediment core-microcosms, when compared on a per g dry weight basis. However, when the results were expressed as O₂ consumed per unit surface area, the values were more similar. CO₂ evolution was less dependent on surface area as evidenced by similar CO₂ values per g sediment in both microcosms. In addition, the effect of sediment mass on O₂ consumption and CO₂ evolution was examined. Both O₂ consumption and CO₂ evolution (expressed as µmole g^–1 dry weight sediment) decreased significantly with increasing sediment mass between 3 and 12 g dry weight. Maximum O₂ consumption per unit headspace was measured when a wet sediment mass between 10.0 and 20.0 g was used in the flask-microcosms. It was also shown that the sediment: water ratio, agitation, incubation temperature, and previous storage time of sediment all affected the respiration estimates. Initial O₂ consumption and CO₂ evolution in flasks were significantly higher in flasks with a decreased sediment: water ratio (1:1 versus 1:2), increased flask agitation, and increased incubation temperature (15 °C versus 5 °C). Also, respiration decreased significantly over the first 100 days of storage at 4 °C.     

Purification of proteins of the Na/Cl cotransporter from membranes of Ehrlich ascites cells using a bumetanide-sepharose affinity column

Summary Bumetanide-binding proteins were isolated from membranes of Ehrlich ascites tumor cells by affinity chromatography. An affinity column was constructed with the active moiety of bumetanide as a ligand using 4-azidobumetanide, a photoactive analogue which inhibits Na/Cl cotransport in Ehrlich cells with high specificity. Covalent binding of the 4-azidobumetanide with Sepharose was promoted by photolysis. Membranes isolated from Ehrlich cells were solubilized withn-octylglucoside. Solubilized proteins retarded by the affinity column were readily eluted by bumetanide. In reducing gels the major proteins eluted by bumetanide were 76 kDa and 38–39 kDa. There were also two proteins of 32 to 35 kDa eluted in lesser amounts. No proteins retarded by the affinity column were eluted with extensive washing without bumetanide. Furthermore, bumetanide eluted no proteins from a control column lacking the specific ligand. Upon rechromatography with bumetanide in solution, bumetanide-eluted proteins were not retarded, but their purity was increased by the retardation of contaminating proteins. Bumetanide-binding protein purified in this manner were characterized further by electrophoresis in nonreducing, nondenaturing gels.     

Draining the Pool? Carbon Storage and Fluxes in Three Alpine Plant Communities

Shrub communities have expanded in arctic and alpine tundra during recent decades. Changes in shrub abundance may alter ecosystem carbon (C) sequestration and storage, with potential positive or negative feedback on global C cycling. To assess potential implications of shrub expansion in different alpine plant communities, we compared C fluxes and pools in one Empetrum-dominated heath, one herb- and cryptogam-dominated meadow, and one Salix-shrub community in Central Norway. Over two growing seasons, we measured Gross Ecosystem Photosynthesis, Ecosystem Respiration (ER), and C pools for above-ground vegetation, litter, roots, and soil separated into organic and mineral horizons. Both the meadow and shrub communities had higher rates of C fixation and ER, but the total ecosystem C pool in the meadow was twice that of the shrub community because of more C in the organic soil horizon. Even though the heath community had the lowest rates of C fixation, it stored one and a half times more C than the shrub community. The results indicate that the relatively high above-ground biomass sequestering C during the growing season is not associated with high C storage in shrub-dominated communities. Instead, shrub-dominated areas may be draining the carbon-rich alpine soils because of high rates of decomposition. These processes were not shown by mid-growing season C fluxes, but were reflected by the very different distribution of C pools in the three habitats.     

Abnormal synthesis of dolichol-linked oligosaccharides in carbohydrate-deficient glycoprotein syndrome

Carbohydrate-deficient glycoprotein syndrome (CDGS) is a raremetabolic disorder presenting in infancy with severe neurologicinvolvement and variable multisystemic abnormalities. Diagnosisrelies upon the detection of abnormal serum glycoprotein isoformson isoelectric focusing (IEF) gels. Carbohydrate structuralanalyses were performed on the N-linked oligosaccharides ofserum     

Tree and Liana Enumeration and Diversity on a One-Hectare Plot in Papua New Guinea1

Enumeration of a one hectare plot at 900 m a.s.l in Papua New Guinea revealed 693 individuals of 228 tree and liana species ≥ 10 cm DBH. A 0.1 hectare subplot contained 302 individuals of 106 species 2.5 ≥ DBH < 10 cm. Lauraceae, Moraceae, and Myristicaceae were the most important families in both size classes. This site is very diverse compared with other tropical forests, and like other species-rich sites worldwide, it has high aseasonal rainfall and high rates of natural disturbance.     

The initiation mess?

This review concerns the mechanisms which control initiation of chromosome replication in enterobacteria with respect to cell growth. Initiation control is commonly separated into positive and negative regulatory mechanisms. Four main points are advanced concerning these different aspects of initiation control. (i) The average concentration of the initiator protein DnaA is proportional to the origin concentration, i.e. the origin per cell mass ratio and, thus, inversely proportional to the very often used term of the 'initiation mass'. (ii) The time of initiation of chromosome replication in the cell cycle is set by DnaA protein accumulating to a threshold level, which in concert with a number of other factors allows for a co-operative formation of the initiation complex. (iii) The time of initiation is not determined by the interaction with these other factors or by the transient interaction between newly replicated origins ( oriC  ) and the cell surface. (iv) The aberrant initiation phenotype observed in various mutants, including dnaA (ts) mutants, might be due to a defective pre-initiation DnaA– oriC interaction or it might be due to a defect in the protection of newly initiated origins from reinitiation. Many of these points are discussed and evaluated in view of recent developments concerning the regulation of chromosome replication in Escherichia coli     

Frequency- and voltage-dependent effects of disopyramide in canine Purkinje fibers

The voltage- and frequency-dependent blocking actions of disopyramide were assessed in canine Purkinje fibers within the framework of concentrations, membrane potentials, and heart rates which have relevance to the therapeutic actions of this drug. Vmax was used to assess the magnitude of sodium channel block. Disopyramide produced a concentration- and rate-dependent increase in the magnitude and kinetics of Vmax depression. Effects on activation time (used as an estimate of drug effect on conduction) were exactly analogous to effects on Vmax. A concentration-dependent increase in tonic block was also observed. Despite significant increases in tonic block at more depolarized potentials, rate-dependent block increased only marginally with membrane potential over the range of potentials in which propagated action potentials occur. Increases in extracellular potassium concentration accentuated drug effect on Vmax but attenuated drug effect on action potential duration. Recovery from rate-dependent block followed two exponential processes with time constants of 689 +/- 535 ms and 15.7 +/- 2.7 s. The latter component represents dissociation of drug from its binding site and the former probably represents recovery from slow inactivation. A concentration-dependent increase in the amplitude of the first component suggested that disopyramide may promote slow inactivation. There was less than 5% recovery from block during intervals equivalent to clinical diastole. Thus, depression of beats of all degrees of prematurity was similar to that of basic drive beats. Prolongation of action potential duration by therapeutic concentrations of drug following a long quiescent interval was minimal. However, profound lengthening of action potential duration occurred following washout of drug effect at a time when Vmax depression had reverted to normal, suggesting that binding of disopyramide to potassium channels may not be readily reversed. Variable effects on action potential duration may thus be attributed to a block of the window current flowing during the action potential being partially or over balanced by block of potassium channels. Purkinje fiber refractoriness was prolonged in a frequency-dependent manner. Disopyramide did not significantly alter the effective refractory period of basic beats but did increase the effective refractory period of sequential tightly coupled extra stimuli. The results can account for the antiarrhythmic actions of disopyramide during a rapid tachycardia and prevention of its initiation by programmed electrical stimulation.