Symplasmic and apoplasmic transport inside feather moss stems of Pleurozium schreberi and Hylocomium splendens.

Background and Aims: The ubiquitous feather mosses Pleurozium schreberi and Hylocomium splendens form a thick, continuous boundary layer between the soil and the atmosphere, and play important roles in hydrology and nutrient cycling in tundra and boreal ecosystems. The water fluxes among these mosses and environmental factors controlling them are poorly understood. The aim of this study was to investigate whether feather mosses are capable of internal transport and to provide a better understanding of species-specific morphological traits underlying this function. The impacts of environmental conditions on their internal transport rates were also investigated. Methods: Cells involved in water and food conduction in P. schreberi and H. splendens were identified by transmission electron microscopy. Symplasmic and apoplasmic fluorescent tracers were applied to the moss stems to determine the routes of internal short- and long-distance transport and the impact of air humidity on the transport rates. Key Results: Symplasmic transport over short distances occurs via food-conducting cells in both mosses. Pleurozium schreberi is also capable of apoplasmic internal long-distance transport via a central strand of hydroids. These are absent in H. splendens. Reduced air humidity significantly increased the internal transport of both species, and the increase was significantly faster for P. schreberi than for H. splendens. Conclusions: Pleurozium schreberi and Hylocomium splendens are capable of internal transport but the pathway and conductivity differ due to differences in stem anatomy. These results help explain their varying desiccation tolerance and possibly their differing physiology and autecology and, ultimately, their impact on ecosystem functioning.

Distant effects of nitric oxide inhalation in lavage-induced lung injury in anaesthetised pigs.

BACKGROUND: Inhalation of nitric oxide (INO) exerts both local and distant effects. INO in healthy pigs causes down-regulation of endogenous nitric oxide (NO) production and vasoconstriction in lung regions not reached by INO, especially in hypoxic regions, which augments hypoxic pulmonary vasoconstriction. In contrast, in pigs with endotoxemia-induced lung injury, INO causes increased NO production in lung regions not reached by INO. The aim of this study was to investigate whether INO exerts distant effects in surfactant-depleted lungs. METHODS: Twelve pigs were anaesthetised, and the left lower lobe (LLL) was separately ventilated. Lavage injury was induced in all lung regions, except the LLL. In six pigs, 40 ppm INO was given to the LLL (INO group), and the effects on endogenous NO production and blood flow in the lavage-injured lung regions were studied. Six pigs served as a control group. NO concentration in exhaled air (ENO), NO synthase (NOS) activity and cyclic guanosine monophosphate (cGMP) in lung tissue, and regional pulmonary blood flow were measured. RESULTS: The calcium (Ca(2+) )-dependent NOS activity was lower (P < 0.05) in the lavage-injured lung regions in the INO group than in the control group. There were no measurable differences between the groups for Ca(2+) -independent NOS activity, cGMP, ENO, or regional pulmonary blood flow. CONCLUSIONS: Regional INO did not increase endogenous NO production in lavage-injured lung regions not directly reached by INO, but instead down-regulated the constitutive calcium-dependent nitric oxide synthase activity, indicating that NO may inhibit its own synthesis.

No effect of metabolic acidosis on nitric oxide production in hypoxic and hyperoxic lung regions in pigs.

AIM: In the severely ill intensive care patients metabolic acidosis and hypoxia often co-exist. We studied the effects of metabolic acidosis on nitric oxide synthase (NOS) dependent and NOS independent nitric oxide (NO) production in hypoxic and hyperoxic lung (HL) regions in a pig model. METHODS: Eighteen healthy anaesthetized pigs were separately ventilated with hypoxic gas to the left lower lobe (LLL) and hyperoxic gas to the rest of the lung. Six pigs received HCl infusion (HCl group), six pigs received the non-specific NOS inhibitor N(ω) -nitro-l-arginine methyl ester (l-NAME) and HCl infusions (l-NAME + HCl group) and six pigs received buffered Ringer's solution (control group). NO concentration in exhaled air (ENO), NOS activity in lung tissue, and regional pulmonary blood flow were measured. RESULTS: Metabolic acidosis, induced by infusion of HCl, decreased the relative perfusion to the hypoxic LLL from 7 (3) [mean (SD)] to 3 (1) % in the HCl group (P < 0.01), and from 4 (1) to 1 (1) % in the l-NAME + HCl group (P < 0.05), without any measurable significant changes in ENO from hypoxic or HL regions There were no significant differences between the HCl and control groups for Ca(2+) -dependent (cNOS) or Ca(2+) -independent NOS (iNOS) activity in hypoxic or HL regions. CONCLUSIONS: Metabolic acidosis augmented the hypoxic pulmonary vasoconstriction, without any changes in pulmonary NOS dependent or NOS independent NO production. When acidosis was induced during ongoing NOS blockade, the perfusion of hypoxic lung regions was almost abolished, indicating acidosis-induced pulmonary vasoconstriction was not NO dependent.

Nitrogen mineralization and phenol accumulation along a fire chronosequence in northern Sweden.

Scots pine (Pinus sylvestris L.) forests of northern Sweden are often considered to be N limited. This limitation may have been exacerbated by the elimination of wildfire as a natural disturbance factor in these boreal forests. Phenolic inhibition of N mineralization and nitrification (due to litter and exudates of ericaceous shrubs) has been proposed as a mechanism for N limitation of these forests, but this hypothesis remains largely untested. N mineralization rates, nitrification rates, and sorption of free phenolic compounds were assessed along a fire-induced chronosequence in northern Sweden. A total of 34 forest stands varying in age since the last fire were identified and characterized. Overstorey and understorey vegetative composition and depth of humus were analysed in replicated plots at all 34 sites. Eight of the forest stands aged 3-352 years since the last fire were selected for intensive investigation in which ten replicate ionic resin capsules (used to assess net N mineralization and nitrification) and non-ionic carbonaceous resin capsules (used to assess free phenolic compounds) were installed at the interface of humus and mineral soil. A highly significant correlation was observed between site age and net sorption of inorganic N to resin capsules. Net accumulation of NH4+ and NO3- on resin capsules followed a linear decrease (R 2=0.61, P<0.01) with time perhaps as a result of increased N immobilization with successional C loading. NO3- sorption to resin capsules followed a logarithmic decrease (R 2=0.80, P<0.01) that may be related to a logarithmic increase in dwarf shrub cover and decreased soil charcoal sorption potential along this chronosequence. A replicated field study was conducted at one of the late successional field sites to assess the influence of charcoal and an added labile N source on N turnover. Three rates of charcoal (0, 100, and 1,000 g M-2) and two rates of glycine (0 and 50 g N as glycine M-2) were applied in a factorial design to microplots in a randomized complete block pattern. Net ammonification (as assessed by NH4+ sorption to resins) was readily increased by the addition of a labile N source, but this increase in NH4+ did not stimulate nitrification. Nitrification was stimulated slightly by the addition of charcoal resulting in similar levels of resin-sorbed NO3- as those found in early successional sites. Resin-sorbed polyphenol concentrations were decreased with charcoal amendments, but were actually increased with N amendments (likely due to decomposition of polyphenols). Net N mineralization appears to be limited by rapid NH4+ immobilization whereas nitrification is limited by the lack of an appropriate environment or by the presence of inhibitory compounds in late successional forests of northern Sweden.

Characterisation of the differential interference effects of two boreal dwarf shrub species.

Our purpose was to characterize the relative competitive and phytotoxic potential of two closely related dwarf-shrub species, Empetrum nigrum and E. hermaphroditum, which form clones in a mosaic pattern in post-fire successions of the boreal forest of northern Sweden. We determined morphological and growth parameters of both species, performed bioassays and chemical analysis, and established field experiments to explore possible differing interference effects on trees by the two species. Both Empetrum species had very similar morphological and growth characteristics. E. hermaphroditum exerted considerably greater negative effects than E. nigrum against Pinus sylvestris and Populus tremula seed and seedlings. These negative effects were related to the different substitution of a bibenzyl in the two species. The effect on seed germination of the two bibenzyls isolated from E. nigrum and E. hermaphroditum was compared with that of other simple phenolics; the latter were found to be inactive, indicating a specific phytotoxic component in the bibenzyls. P. sylvestris seeds planted into clones of both species in the field revealed that E. hermaphroditum had much stronger inhibitory effects than did E. nigrum and the addition of activated carbon partially reversed these effects. Seed germination, biomass and survival of P. sylvestris after four seasons were significantly lower in E. hermaphroditum- than in E. nigrum-dominated plots. We conclude that while both Empetrum species are superficially very similar morphologically, they have vastly different effects on tree seed germination, seedling establishment and growth. These effects appear to be due, at least in part, to the different chemical profile of the two species.

The charcoal effect in Boreal forests: mechanisms and ecological consequences.

Wildfire is the principal disturbance regime in northern Boreal forests, where it has important rejuvenating effects on soil properties and encourages tree seedling regeneration and growth. One possible agent of this rejuvenation is fire-produced charcoal, which adsorbs secondary metabolites such as humus phenolics produced by ericaceous vegetation in the absence of fire, which retard nutrient cycling and tree seedling growth. We investigated short-term ecological effects of charcoal on the Boreal forest plant-soil system in a glasshouse experiment by planting seedlings of Betula pendula and Pinus sylvestris in each of three humus substrates with and without charcoal, and with and without phenol-rich Vaccinium myrtillus litter. These three substrates were from: (1) a high-productivity site with herbaceous ground vegetation; (2) a site of intermediate productivity dominated by ericaceous ground vegetation; and (3) an unproductive site dominated by Cladina spp. Growth of B. pendula was stimulated by charcoal addition and retarded by litter addition in the ericaceous substrate (but not in the other two), presumably because of the high levels of phenolics present in that substrate. Growth of P. sylvestris, which was less sensitive to substrate origin than was B. pendula, was unresponsive to charcoal. Charcoal addition enhanced seedling shoot to root ratios of both tree species, but again only for the ericaceous substrate. This response is indicative of greater N uptake and greater efficiency of nutrient uptake (and presumably less binding of nutrients by phenolics) in the presence of charcoal. These effects were especially pronounced for B. pendula, which took up 6.22 times more nitrogen when charcoal was added. Charcoal had no effect on the competitive balance between B. pendula and P. sylvestris, probably due to the low intensity of competition present. Juvenile mosses and ferns growing in the pots were extremely responsive to charcoal for all sites; fern prothalli were entirely absent in the ericaceous substrate unless charcoal was also present. Charcoal stimulated active soil microbial biomass in some instances, and also exerted significant although idiosyncratic effects on decomposition of the added litter. Our results provide clear evidence that immediately after wildfire fresh charcoal can have important effects in Boreal forest ecosystems dominated by ericaceous dwarf shrubs, and this is likely to provide a major contribution to the rejuvenating effects of wildfire on forest ecosystems.

Prompt epithelial damage and restitution processes in allergen challenged guinea-pig trachea in vivo.

BACKGROUND: Little is known about the induction and the morphology of epithelial damage, and of the ensuing epithelial restitution processes in allergic airways. OBJECTIVE: To examine epithelial damage and restitution in allergen challenged guinea-pig trachea. METHODS: Whole-mount techniques, transmission and scanning electron microscopy, cryosectioning, and histochemical staining were used. Cell proliferation was monitored by BrdU-immunohistochemistry. RESULTS: Allergen challenge produced patchy, crater-like, and leucocyte-rich epithelial damage sites. At 1, 5, and 24 h damage was associated with poorly differentiated epithelial restitution cells. Already at 1 h the epithelial craters had a floor of flattened restitution cells and the damaged areas comprised < 1% of the mucosal surface area (whole-mount preparations). In contrast, cryo sections displayed large areas (approximately 20%, 1 h) of denudation. Epithelial, and subepithelial (fibroblasts, smooth muscle) proliferation was increased 5 and 24 h after challenge (P < 0.01). CONCLUSION: Within 1 h allergen challenge has induced patchy damage sites where epithelial restitution is already advanced; although easily produced by cryosectioning frank denudation was not evident in whole-mount preparations. The present findings may explain the well maintained, functional tightness of allergic airways displaying epithelial damage, shedding, and even denudation. The present data also suggest the possibility that epithelial damage-restitution may be causative to allergic airway remodelling.

Association between inflammation and epithelial damage-restitution processes in allergic airways in vivo.

BACKGROUND: Associations between allergen challenge-induced sites of epithelial damage and the distribution of leucocytes and extravasated plasma remain unexplored. OBJECTIVE: To study neutrophils, eosinophils, and fibrinogen at allergen challenge-induced patchy epithelial damage-restitution sites in guinea-pig trachea. METHODS: After local challenge tracheal tissue (cryo sections and whole-mounts) and lumen (selective tracheal lavage) were examined at 1, 5, and 24 h. Eosinophils, neutrophils and fibrinogen were identified by histochemistry. RESULTS: Neutrophils increased markedly in tracheal lavage fluids and in tissue and were strongly associated with the challenge-induced epithelial craters of damage-restitution. At 1 and 24 h eosinophils were increased in the tracheal lumen whereas the surrounding tissue displayed a reversed pattern. Gels rich in fibrinogen, neutrophils, and eosinophils were present in epithelial crater areas, protruding into the lumen. Clusters of free eosinophil granules, Cfegs, released through lysis of eosinophils, and neutrophils with long cytoplasmatic protrusions abounded in these crater areas. CONCLUSION: The present findings provide important new insights into allergic airways where sites of epithelial damage-restitution processes emerge as the major loci for eosinophil, neutrophil, and plasma protein activities, the latter likely causing leukocyte adhesion and activation in vivo. The distribution of eosinophils in this study suggests roles of these cells both in airway mucosa and in regional lymph nodes. Based on the present study we also propose that lysis of eosinophils and Cfegs generation are a major paradigm for activation of these cells in vivo.

Effects of bilberry (Vaccinium myrtillus L.) litter on seed germination and early seedling growth of four boreal tree species.

Laboratory and greenhouse bioassays were used to test for inhibitory effects of senescent and decomposed leaves and aqueous extract from bilberry (Vaccinium myrtillus L.) against seed germination and seedling growth of aspen (Populus tremula L.), birch (Betula pendula Roth.), Scots pine (Pinus sylvestris L.), and Norway spruce [Picea abies (L.) Karst.]. Aqueous extracts from bilberry leaves were inhibitory to aspen seed germination and seedling growth and also induced root damage and growth abnormalities. Addition of activated carbon removed the inhibitory effects of extracts. Senescent leaves reduced pine and spruce seed germination, but rinsing of seeds reversed this inhibition. Senescent leaves were more inhibitory than decomposed leaf litter, suggesting that the inhibitory compounds in bilberry leaves are relatively soluble and released at early stages during decomposition. Spruce was generally less negatively affected by litter and aqueous extracts than the other tested species. This study indicates that chemical effects of bilberry litter have the potential to inhibit tree seedling recruitment, but these effects were not consistently strong. Phytotoxicity is unlikely to be of critical importance in determining success for spruce seedling establishment.

Seasonal variation in phytotoxicity of bracken (Pteridium aquilinum L. Kuhn).

Laboratory bioassays were used to test for the phytotoxicity of volatile compounds, fresh plant material as a seed bed, and water extracts from bracken [Pteridium aquilinum (L.) Kuhn] pinnules to germination and seedling growth of aspen (Populus tremula L.) and Scots pine (Pinus sylvestris L.). Fronds were sampled from two bracken populations, one in the south and one in the north of Sweden. All three bioassays showed inhibitory effects, and these varied seasonally with the most inhibitory effects occurring in May, June, and September. The peak of inhibition in May and June coincides with the start of the growing season when bracken still is immature and vulnerable to interference from other species. The increase in inhibitory effects in September appears to be due to transformation of natural products or an accumulation of inhibitory compounds that are released during decomposition following frond death. Addition of activated carbon did not remove the inhibitory effects.

Inhibition of Scots pine seedling establishment byEmpetrum hermaphroditum.

Poor establishment and reduced seedling growth of Scots pine (Pinus silvestris L.) in northern Sweden is related to an allelopathic inhibition by the dwarf shrubEmpetrum hermaphroditum Hagerup. Indoor bioassays with green and brown leaves ofEmpetrum have strong negative effects on rooting ability, radicle elongation, and growth of Scots pine seedlings. Bioassays with soil samples show that phytotoxic substances leached fromEmpetrum foliage accumulate in the soil. Field experiments reveal that chemical inhibition byEmpetrum, causing high mortality and slow growth of pine seedlings, can be reduced by adding activated carbon to the soil.