Simultaneous Isolation of Glial and Neuronal Fractions from Rat Brain Homogenates: Comparison of High-Affinity L-Glutamate Transport Properties

An improved three-step Percoll density gradient centrifugation technique is described for simultaneous isolation of glial plasmalemmal vesicles (GPV) and synaptosomal vesicles (SYN) from a rat brain homogenate. While electron microscopy revealed that fractions contained intact vesicles with markedly distinct morphological features, measures of high-affinity [³H]choline uptake, glutamine synthetase and carbonic anhydrase activities, as well as Western blot analyses for glial fibrillary acidic protein and neuron specific enolase, served to confirm the low level of neuronal contamination in GPV fractions as well as the low level of glial contamination in SYN fractions. In addition, GPV and SYN fractions were used to characterize the kinetic and pharmacological properties of sodium-dependent [³H]L-glutamate transport. In conclusion, these results demonstrate the usefulness of this method for obtaining highly-enriched, functionally viable populations of glial and neuronal elements which are suitable for studies of their respective cell functions in vitro.     

Temperature and the catalytic activity of enzymes: A fresh understanding

The discovery of an additional step in the progression of an enzyme from the active to inactive state under the influence of temperature has led to a better match with experimental data for all enzymes that follow Michaelis–Menten kinetics, and to an increased understanding of the process. The new model of the process, the Equilibrium Model, describes an additional mechanism by which temperature affects the activity of enzymes, with implications for ecological, metabolic, structural, and applied studies of enzymes.     

一种改良的对虾白斑综合征病毒的提纯技术

The envelope proteins of White spot syndrome virus (WSSV) are very fragile and easy to be destroyed during purification. It was difficult to obtain a large quantity of intact virions by routine sucrose gradient centrifugation. After modifying the sucrose gradient by adding citrate sodium, we can obtain a large quantity of intact virions and nucleocapsids. This purified virions and nucleocapsids were subsequently used for analyzing viral structural proteins and DNA extraction. The result showed that this modified techniaue is very efficient for virus purification.     

The isolation of Saccharomyces cerevisiae nuclear membranes with nuclease and high-salt treatment

Saccharomyces cerevisiae nuclear membranes were prepared from isolated nuclei by digesting chromatin with deoxyribonuclease and ribonuclease, washing of residual nuclei with 0.5 M MgCl₂, and discontinuous gradient centrifugation in buffered Ficoll solutions. Electron microscopic examination of the preparations showed single membrane and double membrane vesicles and membrane sheets. Pores or residual pores were often visible. In double membrane profiles the two unit membranes were often separated by the remains of the perinuclear cistern. The nuclear membrane fragments contained 58% protein, 23.8% phospholipid, 6% sterols, 7.1% neutral acylglycerols, 4.8% RNA, and 0.3% DNA. The phospholipid content of the membrane preparations was influenced by a phospholipase activity with acidic pH optimum.     

Quasi-elastic light scattering of Artemia ribosomes sedimented in a CsCl density gradient

It is impossible to measure the diffusion coefficient of macromolecules directly and accurately by quasi—elastic light scattering, when aggregates cannot be eliminated from the solutions to be investigated. Nevertheless, a simple method can be applied to overcome this problem in many cases. Aggregates are separated from the monomeric macromolecules by rate-zonal sedimentation in a CsCl density gradient in a transparent centrifugation tube; the monomers are then located by laser light scattering intensity measurements; photon correlation spectroscopy of the scattered light finally yields their diffusion coefficient. The viscosity of aqueous CsCl solutions at different temperatures and concentrations allows a good separation by centrifugation and a low uncertainty in the reduction of the measured diffusion coefficient to standard conditions.The application of the method to eukaryotic large ribosomal subunits is described as an example.     

Is Urografin density gradient centrifugation suitable to separate nonculturable cells from <Emphasis Type="Italic">Escherichia coli</Emphasis> populations?

The ability of Urografin or Percoll density gradient centrifugations to separate nonculturable subpopulations from heterogeneous Escherichia coli populations was analysed. Bacterial counts (total, active and culturable cells) and flow cytometric analyses were carried out in all recovered bands. After Urografin centrifugation, and despite the different origin of E. coli populations, a common pattern was obtained. High-density bands were formed mainly by nonculturable cells. However, the increase in cell density would not be common to all nonculturable cells, since part of this subpopulations banded in low-density zones, mixed with culturable cells. Bands obtained after Percoll centrifugation were heterogeneous and culturable and nonculturable cells were recovered along the gradient. Thus, fractionation in Urografin cannot be only attributed to changes in buoyant densities during the transition from culturable to nonculturable state. Urografin density gradients allow us to obtain enriched fractions in nonculturable subpopulations from a heterogeneous population, but working conditions should be carefully chosen to avoid Urografin toxicity.     

Latitudinal characteristics of below- and above-ground biomass of Typha: a modelling approach

BACKGROUND AND AIMS: The latitudinal differences in the growth characteristics of Typha are largely unknown, although a number of studies have pointed out the effects of climate on the growth and productivity of Typha. Therefore, a dynamic growth model was developed for Typha to examine the effects of latitudinal changes in temperature and radiation on partitioning of the total biomass during the growing season into rhizomes, roots, flowering and vegetative shoots, and inflorescences. METHODS: After validating the model with data from growth studies of Typha found in past literature, it was used to investigate the dynamics of above- and below-ground biomasses at three latitudes: 30 degrees, 40 degrees and 50 degrees. KEY RESULTS: Regardless of the initial rhizome biomass, both above- and below-ground biomass values converged to a latitude-specific equilibrium produced by the balance between the total production and respiration and mortality losses. Above-ground biomass was high from 10 degrees to 35 degrees latitude with sufficient radiation, despite high metabolic losses; however, it decreased markedly at higher latitudes due to a low photosynthetic rate. Below-ground biomass, on the other hand, increased with latitude up to 40 degrees due to decreasing metabolic losses, and then markedly decreased at higher latitudes. Above-ground biomass was enhanced with an increasing number of cohorts regardless of latitude. However, although more cohorts resulted in a larger below-ground biomass at low latitudes, the largest below-ground biomass was provided by a smaller number of cohorts at high latitudes. This difference is due to low production rates of late-season cohorts in high latitudes, compared with consumption for shooting and establishing foliage. CONCLUSIONS: The model could be used to predict the potential growth of Typha in given conditions over a wide range of latitudes and is useful for practical applications such as wetland management or wastewater treatment systems using Typha.     

Species turnover in deciduous forest vegetation

Summary Permanent quadrats were marked out in two areas of hardwood forest vegetation in 1969, and listings of their vascular plant species were taken on several occasions over the snow-free seasons of 1969, 1970, 1971 and 1976. Over the period of study, mean numbers of species per m² remained virtually constant, but variations in the species compositions of individual quadrats were such that mean turnover ratios of 0.115 and 0.085, respectively, were computed for the two stands. Between 1969 and 1976 averages of 20% and 14%, respectively, of the species found in individual quadrats were replaced. This was not accomplished by qualitative changes in the floras of the two stands. Rather, it reflects the operation of a system of continuous rearrangements of species in the small quadrats of both sample areas.Taxonomical nomenclature and life-form system used in this study are according to Gleason & Cronquist (1963).William Phillips, Ian Sutherland and Sheila Thompson helped in the field; Professor Keith Wade commented on the material; Abal Sen drafted the diagram; and the research is part of that funded by the National Research Council of Canada.     

Equilibrium analysis and phase synchronization of two coupled HR neurons with gap junction

The properties of equilibria and phase synchronization involving burst synchronization and spike synchronization of two electrically coupled HR neurons are studied in this paper. The findings reveal that in the non-delayed system the existence of equilibria can be turned into intersection of two odd functions, and two types of equilibria with symmetry and non-symmetry can be found. With the stability and bifurcation analysis, the bifurcations of equilibria are investigated. For the delayed system, the equilibria remain unchanged. However, the Hopf bifurcation point is drastically affected by time delay. For the phase synchronization, we focus on the synchronization transition from burst synchronization to spike synchronization in the non-delayed system and the effect of coupling strength and time delay on spike synchronization in delayed system. In addition, corresponding firing rhythms and spike synchronized regions are obtained in the two parameters plane. The results allow us to better understand the properties of equilibria, multi-time-scale properties of synchronization and temporal encoding scheme in neuronal systems.     

Fractionation of Hevea brasiliensis latex on ficoll density gradients

Fractionation of diluted Hevea brasiliensis latex on iso-osmotic Ficoll density gradients by sedimentation produces a sharp acid phosphatase-rich lutoid band at the density of ca1.04 g/cm³. Polyphenol oxidase (PPO) is not exclusively located in the Frey-Wyssling complexes as shown earlier by others; its distribution is variable. A complete separation of acid phosphatase and PPO activities can be obtained by flotation in a Ficoll density gradient. Ethephon stimulation of the tree reduces the acid phosphatase and PPO activities. A transient change in lutoid density soon after stimulation is observed. The occurrence of some new enzyme activities and their subcellular localization are reported.