The limits of phosphorus removal in wetlands

The phosphorus concentrations exported from wetlands are explored via data and a first order model. The graph of outlet concentration versus areal phosphorus loading is used to display these data and the model. For a given wetland, data and models show that P concentrations show an S curve response to increasing P loadings. The lower plateau is the background concentration and the upper plateau is the inlet concentration. The position of the ascending limb between the two plateaus is positioned differently for different wetlands. Phosphorus (P) removal in wetlands is often typified by a stable decreasing gradient of P concentrations from inlet to outlet, and an accompanying stable decreasing gradient in P assimilation. Limits to removal are inherent in the physical, chemical and biological processes. A lower outlet concentration limit exists because of the P cycle in the un-impacted wetland. The loading at which the outlet concentration departs from background, the lower knee in the loading curve, varies from wetland to wetland. An upper outlet concentration limit is imposed by the source concentration for extremely high inflows. The first order mass balance equation interpolates between these limits. The model gives further insights about performance within an overall envelope. The water carrying capacity of the wetland precludes flows in excess of the hydraulic conveyance capacity, thus limiting the possible P loadings to the system. Conversely, extremely low hydraulic loadings cause the wetland to be dominated by atmospheric additions and losses. The central tendency of inter-system data in the North American Database is shown to be inadequate to draw generalized conclusions about ecosystem processes in an individual wetland. The proposed one gram rule of Richardson, et al. (1997) is shown to be an over-simplification.     

Effect of Trace Levels of Nigericin on Intracellular pH and Acid-Base Transport in Rat Renal Mesangial Cells

Nigericin is an ionophore commonly used at the end of experiments to calibrate intracellularly trapped pH-sensitive dyes. In the present study, we explore the possibility that residual nigericin from dye calibration in one experiment might interfere with intracellular pH (pH _i ) changes in the next. Using the pH-sensitive fluorescent dye 2′,7′-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF), we measured pH _i in cultured rat renal mesangial cells. Nigericin contamination caused: (i) an increase in acid loading during the pH _i decrease elicited by removing extracellular Na⁺, (ii) an increase in acid extrusion during the pH _i increase caused by elevating extracellular [K⁺], and (iii) an acid shift in the pH _i dependence of the background intracellular acid loading unmasked by inhibiting Na-H exchange with ethylisopropylamiloride (EIPA). However, contamination had no effect on the pH _i dependence of Na-H exchange, computed by adding the pH _i dependencies of total acid extrusion and background acid loading. Nigericin contamination can be conveniently minimized by using a separate line to deliver nigericin to the cells, and by briefly washing the tubing with ethanol and water after each experiment. Received: 14 October 1998/Revised: 2 March 1999     

Effect of acute and chronic exercise on plasma amino acids and prolactin concentrations and on [3H]ketanserin binding to serotonin2A receptors on human platelets

The neurotransmitter serotonin (5-hydroxytryptamine, 5-HT) has been shown to modulate various physiological and psychological functions such as fatigue. Altered regulation of the serotonergic system has been suggested to play a role in response to exercise stress. In the present study, the influence was investigated of acute endurance exercise and short-term increase in the amount of training on the concentrations of the 5-HT precursor tryptophan (TRP), of prolactin (PRL) and of branched-chain amino acids (BCAA) in the blood, as well as on the binding of [3H]ketanserin to the serotonin-2A (5-HT2A) receptors on platelets. Nine healthy endurance-trained men were tested the day before (I) and after (II) a 9-day training programme. Samples of venous blood were drawn after an overnight fast and following 5 h of cycling. Fasted and post-exercise plasma concentrations of free TRP, BCAA and free TRP:BCAA ratio did not differ between I and II. A significant decrease of plasma BCAA (P < 0.01) and significant augmentations of plasma free TRP, free TRP:BCAA ratio and PRL (P < 0.01) were found post-exercise. The increase in plasma PRL was smaller in II compared with I. Acute endurance exercise reduced the density of platelet 5-HT2A receptor [3H]ketanserin binding sites at I and II (P < 0.05). The basal density of the binding sites and the affinity of [3H]ketanserin for these binding sites were unaffected by an increase in the amount of training. The present results support the hypothesis that acute endurance exercise may increase 5-HT availability. This was reflected in the periphery by increased concentration of the 5-HT precursor free TRP, by increased plasma PRL concentration, and by a reduction of 5-HT2A receptors on platelets. It remains to be resolved whether these alterations in the periphery occur in parallel with an increase in the availability of 5-HT in the brain.     

Root anchorage of inner and edge trees in stands of Maritime pine (<Emphasis Type="Italic">Pinus pinaster</Emphasis> Ait.) growing in different podzolic soil conditions

Static winching tests were carried out in order to determine the mechanical resistance of Maritime pine to overturning. The tested stands were selected according to podzolic soil conditions: wet Lande, characterised by a shallow ground water table and a hard pan horizon, and dry Lande, with a deeper ground water table and a hard pan absent or broken up. As this soil horizon limits the vertical growth of tree roots, anchorage resistance was investigated with regards to the presence or absence of a hard pan underneath each tree. To determine if mechanical behaviour differed within a stand, trees from inside the stand and edge trees at the border exposed to prevailing winds were also tested. The critical turning moment (TM_crit,total) at the base of the stem was positively related to the variable (H × DBH²) (H, total tree height; DBH, tree diameter). Linear regression analyses between TM_crit,total and (H × DBH²) showed that the presence of a hard pan had no significant effect on anchorage resistance in uprooted trees. Stem failure occurred for 82% of trees on dry Lande when (H × DBH²) < 1 m³. Moreover, stem failure type on dry Lande indicated that trees were better anchored. On soil with a hard pan, edge trees were found to be 20% more resistant to overturning than inner trees. Edge trees differed from inner trees in that the soil-root plate was two times larger and also possessed a larger surface area on the windward side.     

The Effects of Infrared Loading and Water Table on Soil Energy Fluxes in Northern Peatlands

Increased radiative forcing is an inevitable part of global climate change, yet little is known of its potential effects on the energy fluxes in natural ecosystems. To simulate the conditions of global warming, we exposed peat monoliths (depth, 0.6 m; surface area, 2.1 m²) from a bog and fen in northern Minnesota, USA, to three infrared (IR) loading (ambient, +45, and +90 W m^–2) and three water table (–16, –20, and –29 cm in bog and –1, –10 and –18 cm in fen) treatments, each replicated in three mesocosm plots. Net radiation (Rn) and soil energy fluxes at the top, bottom, and sides of the mesocosms were measured in 1999, 5 years after the treatments had begun. Soil heat flux (G) increased proportionately with IR loading, comprising about 3%–8% of Rn. In the fen, the effect of IR loading on G was modulated by water table depth, whereas in the bog it was not. Energy dissipation from the mesocosms occurred mainly via vertical exchange with air, as well as with deeper soil layers through the bottom of the mesocosms, whereas lateral fluxes were 10–20-fold smaller and independent of IR loading and water table depth. The exchange with deeper soil layers was sensitive to water table depth, in contrast to G, which responded primarily to IR loading. The qualitative responses in the bog and fen were similar, but the fen displayed wider seasonal variation and greater extremes in soil energy fluxes. The differences of G in the bog and fen are attributed to differences in the reflectance in the long waveband as a function of vegetation type, whereas the differences in soil heat storage may also depend on different soil properties and different water table depth at comparable treatments. These data suggest that the ecosystem-dependent controls over soil energy fluxes may provide an important constraint on biotic response to climate change.     

Modulation of the histaminergic system and behaviour by alpha-fluoromethylhistidine in zebrafish

The functional role of histamine (HA) in zebrafish brains was studied. Zebrafish did not display a clear circadian variation in brain HA levels. Loading of zebrafish with l-histidine increased HA concentration in the brain. A single injection of the histidine decarboxylase (HDC) inhibitor, alpha-fluoromethylhistidine (alpha-FMH), gave rise to a rapid reduction in zebrafish brain HA. Low HDC activity in the brain after injections verified the effect of alpha-FMH. A reduction in the number of histaminergic fibres but not neurones and an increased expression of HDC mRNA was evident after alpha-FMH. Automated behavioural analysis after alpha-FMH injection showed no change in swimming activity, but abnormalities were detected in exploratory behaviour examined in a circular tank. No significant behavioural changes were detected after histidine loading. The time spent for performance in the T-maze was significantly increased in the first trial 4 days after alpha-FMH injections, suggesting that lack of HA may impair long-term memory. The rostrodorsal telencephalon, considered to correspond to the mammalian amygdala and hippocampus in zebrafish, is densely innervated by histaminergic fibres. These results suggest that low HA decreases anxiety and/or affects learning and memory in zebrafish, possibly through mechanisms that involve the dorsal forebrain.     

Muscle activity reduces soft-tissue resonance at heel-strike during walking

Muscle activity has previously been suggested to minimize soft-tissue resonance which occurs at heel-strike during walking and running. If this concept were true then the greatest vibration damping would occur when the input force was closest to the resonant frequency of the soft-tissues at heel-strike. However, this idea has not been tested. The purpose of this study was to test whether muscle activity in the lower extremity is used to damp soft-tissue resonance which occurs at heel-strike during walking. Hard and soft shoe conditions were tested in a randomized block design. Ground reaction forces, soft-tissue accelerations and myoelectric activity were measured during walking for 40 subjects. Soft-tissue mass was estimated from anthropologic measurements, allowing inertial forces in the soft-tissues to be calculated. The force transfer from the ground to the tissues was compared with changes in the muscle activity. The soft condition resulted in relative frequencies (input/tissue) to be closer to resonance for the main soft-tissue groups. However, no increase in force transmission was observed. Therefore, the vibration damping in the tissues must have increased. This increase concurred with increases in the muscle activity for the biceps femoris and lateral gastrocnemius. The evidence supports the proposal that muscle activity damps soft-tissue resonance at heel-strike. Muscles generate forces which act across the joints and, therefore, shoe design may be used to modify muscle activity and thus joint loading during walking and running.     

Cyanobacterial toxicity and migration in a mesotrophic lake in western Washington, USA

In fall 1997, the toxic cyanobacterium Microcystis aeruginosa was documented in Lake Sammamish (western Washington, U.S.A.) for the first time. Cyanobacterial activity and environmental conditions that may promote toxic cyanobacteria were investigated during summer and fall 1999. Development of toxic Microcystis was hypothesized to be due to runoff of nutrients from the watershed (external loading hypothesis) or from vertical migration of dormant cyanobacteria from the nutrient-rich sediments into the water column (cyanobacterial migration hypothesis). Microcystins were detected using an enzyme-linked immunosorbent assay during late August and early September 1999 despite low cyanobacterial abundance. Microcystin concentrations ranged between 0.19–3.8 g l^–1 throughout the lake and at all depths with the exception of the boat launch where concentrations reached 43 g l^–1. Comparison of the conditions associated with the toxic episodes in 1997 and 1999 indicate that Microcystis is associated with a stable water column, increased surface total phosphorus concentrations (> 10 g l^–1), surface temperatures greater than 22°C, high total nitrogen to phosphorus ratios (> 30), and increased water column transparency (up to 5.5 m). Migration of the cyanobacteria, Microcystis and Anabaena, occurred in both the deep and shallow portions of the lake. Microcystis dominated (89–99%) the migrating cyanobacteria with greater migration from the shallow station. External loading of nutrients due to the large rainfall preceding the 1997 toxic episode may have provided the nutrients needed to fuel that bloom. However, toxic Microcystis occurred in 1999 despite the lack of rain and subsequent external runoff. The migration of Microcystis from the nutrient-rich sediments may have been the inoculum for the toxic population detected in 1999.     

Continuous isolation of plasmid DNA by annular chromatography.

Continuous chromatographic separations, especially of multicomponent mixtures, constitute interesting options for biotechnological downstream processing. Taking the separation of plasmid DNA from clearified lysates on hydroxyapatite as a pertinent example, we discuss the potential of continuous annular chromatography (CAC) in comparison with conventional (preparative) batch chromatography. In CAC the column is realized in the form of a thin (5 mm, height 210 mm) slowly rotating annulus. The performance of such a CAC column is compared to that of an ("analytical") batch column of similar thickness (diameter) and length (4 x 250 mm) and that of a ("preparative") batch column of similar cross-sectional surface area and height (50 x 210 mm). The quality of the obtained plasmid as defined by the appearance of the corresponding agarose gels (native and linearized plasmid), the 260/280 ratio and the biological activity (transient transfection of HEK 293 cells) was found to be identical in all three cases. The yields are also shown to be equivalent. The loading factor is found to be the most decisive parameter for the transfer of a given separation method between the continuous and the batch columns. Under nonoptimized conditions, plate numbers tended to be lower in the continuous compared to the batch columns. This is shown to be largely due to an artifact created by the CAC design (collection of averaged fractions at the outlets) and can be overcome by optimizing the rotation speed. Surprisingly the large batch column consistently gave better plate numbers than either the small batch or the CAC column. Compared to the preparative batch column, wall effects are more pronounced in the CAC (respectively the small diameter batch column), which may translate into better bed stability but conceivably also contributes to an increase in plate height, due to the reduction in bed density usually observed in the proximity of the wall. The CAC is shown to be a powerful approach to continuous chromatography, which allows a direct and straightforward upscale of chromatographic bioseparation methods.