Effect of phospholipases and proteases on the [3H]N6-(R)-phenylisopropyladenosine ([3H]R-PIA) binding to A1 adenosine receptors from pig cerebral cortex.

The effect of phospholipases and proteases on the membrane-bound and solubilized A1 adenosine receptor has been studied. Phospholipids modulate the [3H]N6-(R)-phenylisopropyladenosine binding to A1 adenosine receptors in crude membranes and in soluble preparations, because changes in the phospholipid environment decrease both the binding capacity and the affinity for the ligand. It has become clear that 1) there is co-solubilization of receptor and phospholipids; 2) the phospholipid requirements are different for the coupled and the uncoupled receptor; 3) a net charge in the polar head produced by phospholipase D prevents the agonist binding to the receptor-G protein complex; alternatively, when the whole polar head is removed by phospholipase C the uncoupled receptor is altered; and 4) the protease action upon the receptor suggests that receptor coupled to G protein is more protected by the membrane than the uncoupled receptor. In kinetic experiments performed on membranes it was demonstrated that phospholipase C and trypsin increased the Kd value of the high-affinity state by modifying both k1 and k-1. In contrast they only modified the dissociation constant of the low-affinity state. In conclusion it should be noted that phospholipids play a key role for the binding of R-PIA to A1 adenosine receptor. Also, a different disposition within the membrane of the coupled and uncoupled receptor is encountered.     

Electroporating Fields Target Oxidatively Damaged Areas in the Cell Membrane

Reversible electropermeabilization (electroporation) is widely used to facilitate the introduction of genetic material and pharmaceutical agents into living cells. Although considerable knowledge has been gained from the study of real and simulated model membranes in electric fields, efforts to optimize electroporation protocols are limited by a lack of detailed understanding of the molecular basis for the electropermeabilization of the complex biomolecular assembly that forms the plasma membrane. We show here, with results from both molecular dynamics simulations and experiments with living cells, that the oxidation of membrane components enhances the susceptibility of the membrane to electropermeabilization. Manipulation of the level of oxidative stress in cell suspensions and in tissues may lead to more efficient permeabilization procedures in the laboratory and in clinical applications such as electrochemotherapy and electrotransfection-mediated gene therapy.     

Spatial extent of bird species response to landscape changes: colonisation/extinction dynamics at the community-level in two contrasting habitats

Animal community dynamics in changing landscapes are primarily driven by changes in vegetation structure and ultimately by how species respond to these changes and at which spatial scale. We consider two major components of local community dynamics, species colonisation and extinction. We hypothesise that (1) the optimal spatial extent needed to accurately predict them will differ between these two processes; (2) it will also likely differ from species to species as a result of life history traits differences related to differences in habitat selection and (3) that a species' primary habitat will determine the spatial extent at which it perceives change in vegetation structure. We used data collected over 25 yr in a changing Mediterranean landscape to study bird species local colonisation and extinction patterns in two groups of species typical from two habitats: open farmland and woodland. Vegetation changes were measured at spatial extents ranging from 0.2 to 79 ha. Local species colonisation and extinction estimates were computed using a method accounting for heterogeneity in detection probability among species. We built linear models between local species colonisation/extinction estimates and vegetation changes and examined variations in model quality with respect to the spatial extent at which vegetation changes had been measured. Models for open habitat species showed that colonisation processes operated at the landscape scale (79 ha), while extinction was more tightly linked to local habitat requirements (0.2 ha). Models for woodland species presented a low and constant model quality whatever the spatial extent considered. Our results suggest that the dynamics of the woodland species considered responded to a combination of vegetation changes at several scales and, in particular, to changes in the vertical structure of the vegetation. We highlight the need to explicitly consider spatial extent in studies of habitat selection and of habitat and population dynamics to improve our understanding of the biological consequences of land use changes and guide more effective conservation efforts.     

Characterization of a lactate dehydrogenase inhibitor protein from rabbit skeletal muscle mitochondrial fraction

A lactate dehydrogenase inhibitor protein is isolated from rabbit skeletal muscle crude mitochondrial fraction. The molecular weight of the inhibitor is approximately 20,000 as determined by size exclusion HPLC. The inhibitor isoelectricpH is 5.3 as determined by agarose or polyacrylamide gel isoelectric focusing. The amino acid composition of the inhibitor is given. The presence of the inhibitor gives an acidic characteristic to the alkaline M₄ lactate dehydrogenase isozyme and the lactate dehydrogenase-inhibitor complex is more stable than the enzyme alone.     

Lactate dehydrogenase binding to the mitochondrial fraction and to a mitochondrial inhibitor as a function of the isoenzymatic composition

Purified rabbit skeletal muscle LDH M4 isoenzyme, but not H4 isoenzyme, was observed to bind to either the crude mitochondrial fraction or a mitochondrial inhibitor. Several sources of LDH isoenzymes in which M-type subunits with an alkaline pI are predominant bind to this crude mitochondrial fraction and are inhibited by the mitochondrial inhibitor. Binding and inhibition have also been observed with H-type isoenzymes with a pI near 7. The binding and the inhibition processes did not occur with H-type isoenzymes with an acid pI or with M-type isoenzymes with pI near 6. The binding capacity of LDH to the mitochondrial fraction and to the mitochondrial inhibitor is very similar and depends on the net protein charge and not on whether the subunits are H- or M-type.     

Teasing out biological effects and sampling artifacts when using occupancy rate in monitoring programs

ABSTRACT.   Recent declines in biodiversity stress the need for rigorous and reliable multispecies monitoring programs. A potential weakness of monitoring programs is a reliance on raw counts and the assumption either of complete detection or of constant detection probability for each species, regardless of the sampling situation. Until recently, these assumptions have largely remained untested and, therefore, to help insure accuracy, bird-monitoring programs have depended on standardization of counts and counts of longer duration. We tested the effectiveness of these strategies for providing unbiased occupancy rates using a method designed to accommodate situations where species detection probabilities are less than one and heterogeneous. We tested the effect of potential sources of heterogeneity in detection probability (vegetation structure, wind velocity, cloud cover, date, and time) on occupancy rate estimates of 13 bird species in southern France. We compared adjusted and raw occupancy rates for two sampling durations (10 and 20 min). Differences between raw and adjusted occupancy rates were low even for the shorter count duration, suggesting that standardized long counts should produce reliable estimates of occupancy rates even in the absence of correction by an appropriate method. This enhances the value of past monitoring programs where long standardized counts were used, but with designs that do not allow corrected estimates. However, we found that detection probability was heterogeneous for most species and that vegetation structure was an important source of heterogeneity. The possible effects of habitat on detection probability should be of special concern for long-term monitoring programs conducted in landscapes where habitats vary across time or space.     

An in situ enclosure experiment to test the solar UVB impact on plankton in a high-altitude mountain lake. I. Lack of effect on phytoplankton species composition and growth

The effect of solar UVB radiation on the growth and speciescomposition of phytoplankton from a high-mountain lake (2417m a.s.l.) was studied in situ for 16 days in two enclosuresof 1 m receiving either full sunlight or sunlight without UVB.A total of 20 species were identified in both enclosures, consistingmainly of dinoflagellates, chrysophytes and diatoms. Duringthe experiment, there were no significant differences in phytoplanktonspecies composition between the two enclos ures. In both treatments,the abundance of phytoplankton increased continuously, and chlorophylla changed by -5-fold. We observed high fluctuations in the abundanceof several species. However, these fluctuations occurred inboth enclosures, and hence they were not related to UVB radiation.Some species were affected by daily fluctuations of radiation(UVA + photosynthetically active radi ation). Cyclotella aff.gordonensis showed a robust positive correlation, whereas speciesof Gymno dinium were negatively correlated. For most species,the sensitivity to radiation found during the experiment wasconsistent with their diurnal vertical distribution in the lake.Our findings suggest that the phytoplankton from this very clearlake (10% of the surface UVB radiation at 305 nm reached 9.6m depth) were well adapted to the high UVB radiation charactensticof high-elevation sites.