Myelin Gene Expression in Immortalized Schwann Cells: Relationship to Cell Density and Proliferation

Abstract: Myelin gene expression was investigated in the immortalized S16 Schwann cell line grown in the presence and absence of serum and at different densities. Protein expression was monitored by western blotting, and message levels were determined by RNase protection assays. To study cell proliferation rates at different cell densities and serum conditions. [³H]thymidine uptake assays and cell counts were performed. Although serum deprivation decreased cell proliferation as expected, the proliferation of S16 cells was unchanged or slightly increased at high density under the conditions of our experiments in either serum-containing or serum-free medium. This increased cell division at high density appeared to be due to greater release of an autocrine growth factor to the medium by dense cell populations. For both sparse and dense cells, substantially more P₀ glycoprotein (P₀) and myelin-associated glycoprotein (MAG) per milligram of total cellular protein were expressed when the cells were proliferating slowly in defined medium in comparison with more rapidly proliferating cells in serum-containing medium. Furthermore, in both serum-containing and defined media, dense cell populations expressed more MAG and P₀ than sparse ones. P₀ mRNA and MAG mRNA levels generally paralleled protein levels. The level of mRNA for peripheral myelin protein-22 (PMP-22) was also increased at high cell density but did not change much when proliferation was decreased by serum deprivation. PMP-22 protein was not detected under any of the growth conditions. The changes in expression of these genes with growth conditions may be specific for myelin proteins, because the expression of a nonmyelin glycoprotein, L1, remained constant. The level of cyclic AMP in the cells did not change with the different growth conditions tested. The results indicate that the S16 Schwann cell line mimics primary or secondary Schwann cells by down-regulating myelin gene expression when it proliferates more rapidly in the presence of serum. Furthermore, in both the presence and absence of serum, there was greater expression of myelin genes at high cell density that was not associated with a decreased proliferative rate. Because evidence for a role of secretory factors in affecting myelin gene expression was not obtained by treating sparse S16 cells with medium conditioned by dense S16 cells, the results suggest that the higher expression of myelin genes at high density may be mediated by cell-to-cell contact.     

Effect of density on age-specific mortality in Drosophila: a density supplementation experiment

Age-specific mortality rates were studied at two adult density levels in four inbred lines of Drosophila melanogaster. In experimental populations, adult densities were maintained at constant levels throughout the experiment by replacing dead flies with live, marked mutants. In control populations, densities declined naturally as the cohorts aged. For all experimental populations the best mortality model is the two-stage Gompertz model, with slower mortality acceleration at older ages. Flies in the experimental populations generally lived longer than flies in control populations, regardless of sex, genotype, or initial density level. The data demonstrate that deceleration of age-specific mortality rates at older ages is not caused by declining cohort densities. Mortality deceleration is a real phenomenon that raises serious questions about the evolution of senescence.     

Stratification of density in dry deciduous forest using satellite remote sensing digital data—An approach based on spectral indices

Forest density expressing the stocking status constitutes the major stand physiognomic parameter of Indian forest. Density and age are often taken as surrogate to structural and compositional changes that occur with the forest succession. Satellite remote sensing spectral response is reported to provide information on structure and composition of forest stands. The various vegetation indices are also correlated with forest canopy closure. The paper presents a three way crown density model utilizing the vegetation indices viz., advanced vegetation index, bare soil index and canopy shadow index for classification of forest crown density. The crop and water classes which could not be delineated by the model were finally masked from normalized difference vegetation index and TM band 7 respectively. The rule based approach has been implemented for land use and forest density classification. The broad land cover classification accuracy has been found to be 91.5%. In the higher forest density classes the classification accuracy ranged between 93 and 95%, whereas in the lower density classes it was found to be between 82 and 85%.     

The presence of inhibitors of the reproduction of Varroa jacobsoni Oud. (Gamasida: Varroidae) in infested cells

The mite Varroa jacobsoni was reared in artificial gelatin cells under laboratory conditions and the possible presence of factors inhibiting Varroa reproduction was studied. In cells infested with three mites, the mean offspring per female was reduced to 75% of that in singly infested cells. When gelatin cells were used for two successive rearing cycles, both the proportion of reproducing females and the offspring per reproducing female were significantly lower in cells that had contained an infested larva during the first rearing cycle than in those with an uninfested larva. The mean reduction of the offspring per female was 48%; this suggests that inhibitors of the reproduction are released into infested cells. Treatment of gelatin cells with the hexane extract of cells in which an infested bee pupa had developed caused a 21% reduction in the mean offspring per female, with a difference close to the significance level (p=0.07).     

13C discrimination by fossil leaves during the late-glacial climate oscillation 12-10 ka BP: measurements and physiological controls

The late-glacial climatic oscillation, 12-10 ka BP, is characterised in ice core oxygen isotope profiles by a rapid and abrupt return to glacial climate. Recent work has shown that associated with this cooling was a drop in atmospheric CO₂ concentration of ca. 50 ppm. In this paper, the impact of these environmental changes on ¹³C discrimination is reported, based on measurements made on a continuous sequence of fossil Salix herbacea leaves from a single site. The plant responses were interpreted using an integrated model of stomatal conductance, CO₂ assimilation and intercellular CO₂ concentration, influenced by external environmental factors. According to the model, temperature exerts a marked influence on ¹³C discrimination by leaves and the pattern of ¹³C changes recorded by the fossil leaves is consistent with other palaeotemperature curves for 12-10 ka BP, particularly the deuterium isotope record from Alaskan Salix woods, which generally reflects ocean temperatures. The gas exchange model correctly accounts for these changes and so permits the reconstruction of ancient rates of leaf CO₂ uptake and loss of water vapour in response to the abrupt late-glacial changes in global climate and CO₂. The approach provides the required physiological underpinning for extracting quantitative estimates of past temperatures and for contributing an ecophysiological explanation for changes in ¹³C discrimination in the fossil record.     

Far-red enrichment and photosynthetically active radiation level influence leaf senescence in field-grown sunflower

Basal leaves frequently senesce before anthesis in high population density crops. This paper evaluates the hypothesis that quantitative and qualitative changes in the light environment associated with a high leaf area index (LAI) trigger leaf senescence in sunflower ( Helianthus annuus L.) canopies. Mean leaf duration (LD, time from achievement of maximum leaf area) of leaf 8 was significantly ( P < 0.05) reduced from 51 to 19 days as crop population density was increased from 0.47 to 4.76 plants m⁻². High compared to low plant population density was associated with earlier reduction in the photosynthetically active radiation (PAR) and red/far-red ratio (R/FR) reaching the target leaf. However the changes in R/FR preceded those in PAR. When the light environment of individual leaves of isolated plants growing under field conditions was manipulated using filters and FR-reflecting mirrors, LD was positively and linearly related with the mean daily PAR (MDR) received in the FR- (no FR enrichment) treatments. FR enrichment of light reaching the abaxial surface of the leaf significantly ( P < 0.05) reduced LD by 9 days at intermediate PAR levels with respect to FR-controls, but did not affect LD at the maximum PAR used in these experiments. However, when light reaching both leaf surfaces was enriched with FR, LD (for leaves receiving maximum PAR) was 13 days shorter than that of the FR- control. These results show that basal leaf senescence in sunflower is enhanced both by a decrease in PAR and by a decrease in R/FR.     

Root,shoot and soil parameters required for process-oriented models of crop growth limited by water or nutrients

A review is given of the prospects for using process-oriented models of water and nutrient uptake in improving integrated agriculture. Government-imposed restrictions on the use of external inputs will increase the likelihood of (temporary) nutrient or water stress in crop production in NW Europe and thus a better understanding is required of shoot-root-soil interactions than presently available. In modelling nutrient and water uptake, three approaches are possible: 1) models-without-roots, based on empirically derived efficiency ratios for uptake of available resources, 2) models evaluating the uptake potential of root systems as actually found in the field and 3) models which also aim at a prediction of root development as influenced by interactions with environmental factors. For the second type of models the major underlying processes are known and research can concentrate on model refinement on the one hand and practical application on the other. The main parameters required for such models are discussed and examples are given of practical applications. For the third type of models quantification of processes known only qualitatively is urgently needed.     

Predicting badger sett numbers: evaluating methods in East Sussex

Abstract. One way in which a species' numbers may be estimated without direct counting is to predict their dispersion and density from more readily available habitat measures, such as landscape variables measured from maps or vegetation variables measured in the field. We compare the power of ordination and regression techniques for predicting badger ( Meles meles L.) numbers at a local scale, using a land class system, map-read landscape variables and field-derived vegetation variables. Sett density was used as a surrogate of badger density. Multiple linear regression using vegetation and landscape variables together gave the most accurate prediction of sett density, while ordination techniques were of lesser value. The addition of vegetation variables to landscape variables did not substantially improve the power of ordination. Outlier Sett Density was predicted more accurately, and by different variables, to Main Sett Density. The relationship between badger ecology and habitat variables that were useful in predicting sett density is discussed.     

Mutual Contact of Adherent Polymorphonuclear Leukocytes Inhibits Their Generation of Superoxide

Superoxide generation by polymorphonuclear leukocytes (PMNs) in suspension, or adherent to glass or plastic, after stimulation with /V-formylmethionyl-leucyl-phenylalanine or phorbol myristate acetate was measured by cytochromec reduction and spin trapping. Amounts of superoxide generated by adherent PM Ns were inversely related to cell density. The generation of hydrogen peroxide was also inhibited at higher cell densities. In contrast to adherent cells, superoxide released by PMNs in suspension linearly increased with respect to cell number over a wider range. Microscopic observation indicated that the number of cells in mutual contact increased rapidly at cell densities higher than 4 × 10⁴ cells/cm², and inhibition of superoxide became apparent at higher cell densities. Mediators which could be released by PMNs, such as NO and adenosine, were not the cause of inhibition. Thesedatu suggest that mutual contact of PMNs suppresses their generation of superoxide. Survival rates of PMNs after stimulation increased at higher densities, indicating that the mutual contact-induced inhibition of superoxide generation by PMNs may be physiologically relevant at sites of inflammation.