Hormone-responsive cells derived from human dental papilla: Characterization in vitro and in vivo in diffusion chambers

Summary Cells of the dental papilla are capable of odontoblastic, fibroblastic, and endothelial differentiation and formation of dentin and the dental pulp. In the present study dental papilla cells, obtained from human tooth buds (HDP cells), were cultured in vitro through 3 to 7 passages. After exposure to prostaglandin E₂ there was a marked decrease in intracellular cyclic AMP (cAMP) levels as compared to hormone-free controls. Parathyroid hormone and calcitonin had stimulatory effects with 1 and 2 log increases in cAMP, respectively. The HDP cells showed moderate activity of alkaline phosphatase, 1 log higher than that of hamster kidney fibroblasts (BHK 13) and 1 log lower than that of osteoblastic osteosarcoma cells (ROS 17/2). When cultured for 4 or 8 wk in diffusion chambers (DC) implanted in athymic mice, many of the HDP cells underwent odontoblastic morphodifferentiation with very long, single processes extending into the matrix. This matrix contained banded and unbanded collagen fibers. Neither light nor electron microscopy of the DC content revealed mineral deposits. These results suggest that HDP cells have an intrinsic potential for partial odontoblastic differentiation; inductive signals like those originating from odontogenic epithelium are probably essential for the completion of hard tissue formation.     

Deep soil heterogeneity and fine root distribution in forests and pastures of eastern Amazonia

Little is known about deep soil heterogeneity, or its relationship with fine root distribution. Beneath a mature, closed-canopy forest of eastern Amazonia, and the pastures and secondary forests that are derived from this forest, soil soft spots and hollow chambers occur to at least 9 meters depth. We measured the vertical distribution of these soil patches, and compared chemical characteristics, mycorrhizal infection, and root density of soil soft spots with the surrounding matrix of more homogeneous soil. Soil soft spots and chambers varied little with depth, but occupied the greatest soil volume (0.8 to 1.2%) from 4 to 6 m depth in the mature forest. Soft spots had lower pH, P availability and arbuscular mycorrhizal infection, and higher K availability than surrounding soil. Root length density was 2 to 15 times higher in soft spots than in surrounding soil. In the pastures, roots were found only in soil soft spots at depths of >3 m. Pastures and secondary forest had more soil chambers in the upper meter of soil than mature forest, but were otherwise indistinguishable in their patterns of deep soil heterogeneity. Soil soft spots may be vestiges of cutter ant nest chambers, while hollow chambers are cutter ant chambers and root channels. Chambers may act as conduits for root penetration and water penetration to deep soil.Abbreviations AM arbuscular mycorrhizae - RLD root length density (root length per unit of soil volume)     

Cultures of bovine tracheal epithelium with differentiated ultrastructure and ion transport

Summary Tracheal epithelial cells were grown on Nuclepore filters coated with human placental collagen. When grown immersed in medium containing fetal bovine serum, cells displayed an undifferentiated ultrastructure (no cilia and a cell height of ∼ 10 μm). Short-circuit current (I_sc) was approximately 1/10 that of the native epithelium. By contrast, when grown in hormonally defined, serum-free medium with an air interface, cells showed I_sc equal to or greater than the original tissue, possessed cilia, and had a cell height of ∼ 50 μm. Responses in I_sc to mediators were similar to those of the original tissue, but differed from those of dog or human tracheal epithelium. Given the ready availability and low cost of the native tissues, bovine tracheal cultures grown in serum-free medium with an air interface should prove useful in studies of airway epithelial physiology.     

Preservation and computer-aided microscopic analysis of planktonic Protozoa and algae

Samples of microplankton and larger nanoplankton (5 to 200 m) are preserved with a combination of Lugol's solution and DaFano's fixative. Organisms are then settled on a gelatin-coated slide, dried and embedded in 40 percent glycerin. Counting and sizing is performed under a microscope using a drawing tube, which facilitates measuring the organisms with a microcomputer-interfaced caliper. An interactive computer program, written in BASIC, allows for estimating the volumes of cells in up to 40 shape/species categories. The program then saves data on a disk, retrieves them, and calculates the results either for individual species (abundance, biomass, and mean cell volume) or as a pooled size spectrum of all organisms measured.     

Direct and indirect effects of elevated CO2 on transpiration from Quercus myrtifolia in a scrub-oak ecosystem

Elevated atmospheric carbon dioxide (C_a) usually reduces stomatal conductance, but the effects on plant transpiration in the field are not well understood. Using constant‐power sap flow gauges, we measured transpiration from Quercus myrtifolia Willd., the dominant species of the Florida scrub‐oak ecosystem, which had been exposed in situ to elevated C_a (350 µmol mol ^{? 1} above ambient) in open‐top chambers since May 1996. Elevated C_a reduced average transpiration per unit leaf area by 37%, 48% and 49% in March, May and October 2000, respectively. Temporarily reversing the C_a treatments showed that at least part of the reduction in transpiration was an immediate, reversible response to elevated C_a. However, there was also an apparent indirect effect of C_a on transpiration: when transpiration in all plants was measured under common C_a, transpiration in elevated C_a‐grown plants was lower than that in plants grown in normal ambient C_a. Results from measurements of stomatal conductance (g_s), leaf area index (LAI), canopy light interception and correlation between light and g_s indicated that the direct, reversible C_a effect on transpiration was due to changes in g_s caused by C_a, and the indirect effect was caused mainly by greater self‐shading resulting from enhanced LAI, not from stomatal acclimation. By reducing light penetration through the canopy, the enhanced self‐shading at elevated C_a decreased stomatal conductance and transpiration of leaves at the middle and bottom of canopy. This self‐shading mechanism is likely to be important in ecosystems where LAI increases in response to elevated C_a.     

Effects of elevated atmospheric CO2 on net ecosystem CO2 exchange of a scrub–oak ecosystem

We report the results of a 2‐year study of effects of the elevated (current ambient plus 350 μmol CO₂ mol^?1) atmospheric CO₂ concentration (C_a) on net ecosystem CO₂ exchange (NEE) of a scrub–oak ecosystem. The measurements were made in open‐top chambers (OTCs) modified to function as open gas‐exchange systems. The OTCs enclosed samples of the ecosystem (ca. 10 m² surface area) that had regenerated after a fire, 5 years before, in either current ambient or elevated C_a. Throughout the study, elevated C_a increased maximum NEE (NEE_max) and the apparent quantum yield of the NEE (φ_NEE) during the photoperiod. The magnitude of the stimulation of NEE_max, expressed per unit ground area, was seasonal, rising from 50% in the winter to 180% in the summer. The key to this stimulation was effects of elevated C_a, and their interaction with the seasonal changes in the environment, on ecosystem leaf area index, photosynthesis and respiration. The separation of these factors was difficult. When expressed per unit leaf area the stimulation of the NEE_max ranged from 7% to 60%, with the increase being dependent on increasing soil water content (W_soil). At night, the CO₂ effluxes from the ecosystem (NEE_night) were on an average 39% higher in elevated C_a. However, the increase varied between 6% and 64%, and had no clear seasonality. The partitioning of NEE_night into its belowground (R_below) and aboveground (R_above) components was carried out in the winter only. A 35% and 27% stimulation of NEE_night in December 1999 and 2000, respectively, was largely due to a 26% and 28% stimulation of R_below in the respective periods, because R_below constituted ca. 87% of NEE_night. The 37% and 42% stimulation of R_above in December 1999 and 2000, respectively, was less than the 65% and 80% stimulation of the aboveground biomass by elevated C_a at these times. An increase in the relative amount of the aboveground biomass in woody tissue, combined with a decrease in the specific rate of stem respiration of the dominant species Quercus myrtifolia in elevated C_a, was responsible for this effect. Throughout this study, elevated C_a had a greater effect on carbon uptake than on carbon loss, in terms of both the absolute flux and relative stimulation. Consequently, for this scrub–oak ecosystem carbon sequestration was greater in the elevated C_a during this 2‐year study period.     

Assessing the impact of ozone on photosynthesis of European beech (Fergus sylvatica L.) in environmental chambers

An exposure — response study with proportionalto-ambient ozone levels was conducted in closed chambers on 3-year-old European beech (Fagus sylvatica L.) of montane origin. The fumigation started in April 1990 and lasted for a single growing season. Climate data and ozone concentrations monitored at an experimental station of the Institute for Applied Plant Biology, Schönenbuch, Switzerland were simulated in the exposure chambers 12 days later (1*O₃). To test exposure-response relations three additional treatments were applied, subambient (0.2*O₃) and two proportionally increased ozone treatments (1.5*O₃ and 2*O₃). The photosynthetic behaviour of the trees in August revealed the light reactions to be less affected than parameters which are related to the dark reactions of photosynthesis. Assimilation (A₃₅₀), apparent carboxylation efficiency (CE), and maximum photosynthetic capacity (A₂₅₀₀) were reduced with increasing ozone concentration. For the ozone response of CE and A₂₅₀₀ Critical Levels were calculated.