Flooded meadow communities an analysis of their productivity in a wet year

The productivity of three plant communities differeing in moisture conditions was studied in the river basin of the Dyje near the village of Lan?hot (Southern Moravia). The communities were as follows:Serratuleto-Festucetum commutatae Balátová-Tulá?ková 1963,Gratiola officinalis—Carex praecox-suzae subass.Galium boreale Balátová-Tulá?ková 1963, andGratiola officinalis—Carex praecox-suzae subass.Rorippa silvestris Balátová-Tulá?ková 1963. The associationGratiola officinalis—Carex praecox-suzae subass.Galium boreale appeared as the most productive one, its biomass maximum W=400 g . m^?2 and the maximum R=0.042 g . g^?1 . day^?1 C=4.84 g . m^?2 . day^?1. Owing to extreme moisture conditions in the year 1966, the associationSerratuleto-Festucetum commutatae was also highly productive, as it reached the following maximum of dry matter production: 240 g . m^?2, R=0.0388g . g^?1 . day^?1 C=4.64 g . m^?2 . day^?1. The maximum value of dry matter in the associationGratiola officinalis—Carex praecox-suzae subass.Rorippa silvestris was 220 g.m^?2. Changes in dry matter production of shoots were evaluated statistically. The dry matter in the underground parts of plants in 1 square metre, collected from the 0–25 cm layer varied from 1,000 to 2,000 g.m^?2. Together with records of the increasing dry matter in the shoots the author kept records of the properties of dead material.     

Mechanisms of cell volume regulation in the proximal segment of the Malpighian tubule of Rhodnius neglectus

The cell volume regulation of the lower segment cells of the Malpighian tubule of Rhodnius neglectus in anisosmotic media was evaluated by using videooptic techniques. When the medium osmolality was increased with addition of 100 mm mannitol the cells shrank to a minimum of 16.84±2.62% and subsequently swelled towards their initial volume undergoing a typical regulatory volume increase (RVI). Replacement of either K⁺ or Cl^? or HCO ₃ ^? by Na⁺, gluconate and phosphate, respectively, abolished the RVI response. Furthermore, the substitution of Na⁺ by tetramethylammonium (TMA⁺) in isosmotic conditions led to cellular swelling and death. Addition of either amiloride 10^?4 m, anthracene-9-COOH 5×10^?4 m, furosemide 5×10^?4 m or ethacrynic acid 5×10^?5 m, also abolished RVI. On the other hand, addition of either Ba²⁺ 10^?3 m, SITS 5× 10^?4 m, ouabain 10^?3 m or vanadate 10^?3 m, did not change the RVI response. When the tubules were incubated in hyperosmotic media with EGTA 2 mm or verapamil 10^?6 m, the RVI response was abolished. In contrast, a decrease of NaCl concentration from 129 to 79 mm induced a cell swelling to a maximum of 33.11+1.73%, but the cells maintained swollen, only partially regulating their volume. These results show that the proximal cells of Malpighian tubule of R. neglectus are able to regulate their volume in hyperosmotic but only partially regulating in hyposmotic solutions. The mechanisms in RVI involve Na⁺, K⁺, Cl^?, Ca²⁺ and HCO ₃ ^? transport pathways and a ouabain-insensitive ATPase stimulated by Na⁺. This work was supported by grants from the Fundação de Amparo à Pesquisa do Estado de São Paulo—FAPESP; Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq e Financiadora de Projetos e Pesquisas-FINEP.     

Uptake of amino acids by actidione-treated yeast cells

The active uptake ofl-aspartic acid, glycine andl-lysine by actidione-treated cells ofSaccharomyces cerevisiae was found to be inhibited by anaerobic conditions in the absence of a source of energy, only facilitated diffusion persisting. Similarly, metabolic inhibitors (iodoacetamide, sodium fluoride and potassium sorbate) inhibited the uptake very substantially. 2,4-Dinitrophenol and sodium azide appeared to inhibit the movement of the transport carrier itself, while uranyl ions showed a complex interaction pattern, ranging from inhibition at concentrations of 10^?6–10^?4 m, to stimulation at concentrations of 3×10^?4–10^?3 m, to pronounced inhibition at higher concentrations. The uptake was pH-dependent with optima forl-aspartic acid near pH 4, for glycine near pH 5, forl-lysine near pH 6.5.     

Live and dead underground plant biomass in a natural meadow hydrosere

In seven meadow stands of natural hydroseres the average total underground biomass ranged from 1.6 to 3.7 kg. m^?2. The highest percentages of live roots (46–60%) and live underground biomass (1.40–1.61 kg. m^?2) were recorded in stands situated around the middle of the hydroseres and the lowest values (18%; 0.48 kg. m^?2 and 23%; 0.86 kg. m^?2) were both on the wettest and the driest sites. The total underground/aboveground biomass ratios varied between 3.2 and 12.2 while the live underground/aboveground biomass ratios lay between 0.7 and 3.4.     

Characterization of uptake and release processes ford- andl-aspartate in primary cultures of astrocytes and cerebellar granule cells

The uptake ofl-andd-aspartate was studied in astrocytes cultured from prefrontal cortex and in granule cells cultured from cerebellum. A high affinity uptake system forl- andd-aspartate was found in both cell types, and the two stereoisomers exhibited essentially the sameK _m - andV _max -values in bouth astrocytes (l-aspartate:K _m 77 μM;V _max 11.8 nmol×min^?1×mg^?1;d-aspartate:K _m 83 μM;V _max 14.0 nmol×min^?1×mg^?1) and granule cells (l-aspartate:K _m 32 μM;V _max 2.8 nmol ×min^?1×mg^?1;d-aspartate:K _m 26 μM;V _max 3.0 nmol×min^?1×mg^?1). To investigate whetherl-glutamate,l-aspartate andd-aspartate use the same uptake system a detailed kenetic analysis was performed. The uptake kinetics of each one of the three amino acids was studied in the presence of the two other amino acids, and no essential differences between the uptake characteristics of the amino acids were found. In addition to the uptake studies the release ofD-aspartate from cerebellar granule cells was investigated and compared withl-glutamate release. A Ca²⁺-dependent, K⁺-induced release was found for both amino acids.     

Homofermentative production of d-lactic acid from sucrose by a metabolically engineered Escherichia coli

Escherichia coli W, a sucrose-positive strain, was engineered for the homofermentative production of d-lactic acid through chromosomal deletion of the competing fermentative pathway genes (adhE, frdABCD, pta, pflB, aldA) and the repressor gene (cscR) of the sucrose operon, and metabolic evolution for improved anaerobic cell growth. The resulting strain, HBUT-D, efficiently fermented 100?g?sucrose?l^?1 into 85?g?d-lactic acid?l^?1 in 72–84?h in mineral salts medium with a volumetric productivity of ~1?g?l^?1?h^?1, a product yield of 85?% and d-lactic acid optical purity of 98.3?%, and with a minor by-product of 4?g?acetate?l^?1. HBUT-D thus has great potential for production of d-lactic acid using an inexpensive substrate, such as sugar cane and/or beet molasses, which are primarily composed of sucrose.     

Effect of winter fire on primary productivity and nutrient concentration of a dry tropical savanna

Burning increased the mean annual canopy and belowground biomass of a dry tropical savanna by 40% and 12%, respectively, while littermass was reduced by 85% in comparison to control savanna. Mean annual aboveground and belowground net primary production were 471 and 631 g m^-2 in control, and 584 and 688 g m^-2 in burned savanna, respectively. Fire caused an increase in mean aboveground net production of 24% and in belowground net production of 9%.Concentration of carbon, nitrogen and phosphorus in vegetation of unburned plots ranged between 34.01–38.59%, 0.85–1.53% and 0.04–0.11% and in soil from 0.95–1%, 0.011–0.13% and 0.017–0.02%, respectively. Fire increased the mean concentrations of N and P by 16% and 42% in vegetation and 18.18% and 17.65% in soil, respectively. Thus winter fire can be an important tool for the management of dry tropical savanna with respect to biomass production and nutritive quality.     

Biomass partitioning in twoCalamagrostis villosa stands on deforested sites

Calamagrostis villosa stands occurring in areas deforested by air-pollution impact in the Moravian-Silesian Beskydy Mountains were characterized by a high dry mass of total underground biomass (3 300 g. m^?2—the slope site, 2 850 g. m^?2—the flat site). The percentage of living roots and rhizomes in total underground biomass was very high (about 70%). The total aboveground biomass was respectively, 321 g.m^?2 (the slope site) and 726 g. m^?2 (the flat site). In unstabilized habitats on steep slope, the higher plant biomass produced was allocated to a more developed root system.     

Small-conductance chloride channels in human peripheral T lymphocytes

During whole-cell patch-clamp recording from normal (nontransformed) human T lymphocytes a chloride current spontaneously activated in >98% of cells (n > 200) in the absence of applied osmotic or pressure gradients. However, some volume sensitivity was observed, as negative pressure pulses reduced the current. With iso-osmotic bath and pipette solutions the peak amplitude built up (time constant ≈23 sec at room temperature), a variable-duration plateau phase followed, then the current ran down spontaneously (time constant ≈280 sec). The anion permeability sequence, calculated from reversal potentials was I^?, Br^? > NO ₃ ^? , Cl^? > CH₃SO ₃ ^? , HCO ₃ ^? > CH₃COO^? > F^? > aspartate, gluconate, SO ₄ ^2? and there was no measurable monovalent cation permeability. The Cl^? current was independent of time during long voltage steps and there was no evidence of voltage-dependent gating; however, the current showed intrinsic outward rectification in symmetrical Cl^? solutions. The conductance of the channels underlying the whole-cell current was calculated from fluctuation analysis, using power-spectral density and variance-vs.-mean analysis. Both methods yielded a single channel conductance of about 0.6 pS at ?70 mV (close to the normal resting potential of T lymphocytes). The power spectral density function was best fit by the sum of two Lorentzian functions, with corner frequencies of 30 and 295 Hz, corresponding to mean open times of 0.54 and 5.13 msec. The pharmacological profile included rapid block by external application of flufenamic acid (50 μm), 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB, 100 μm, [6,7-dichloro-2-cyclopentyl-2,3-dihydro-2-methyl-1-oxo-1H-inden-5-y1) oxy] acetic acid (IAA-94, 250 μm) or 100 μm 1,9-dideoxyforskolin. The stilbene derivatives DIDS (4,4′-diisothiocyano-2,2′ di-sulphonic acid stilbene, 500 μm) and SITS (4-acetamido-4′-isothiocyano-2, 2′-disulphonic acid stilbene, 500 μm) prevented buildup of Cl^? current after a 30-min preincubation at 500 μm. When tested in a mitogenic assay, DIDS, flufenamic acid, NPPB and IAA-94 all inhibited T-cell proliferation, suggesting a physiological function in addition to the observed volume sensitivity.     

Characterization of a d-psicose-producing enzyme, d-psicose 3-epimerase, from Clostridium sp.

The gene coding for d-psicose 3-epimerase (DPEase) from Clostridium sp. BNL1100 was cloned and expressed in Escherichia coli. The recombinant enzyme was purified by Ni-affinity chromatography. It was a metal-dependent enzyme and required Co²⁺ as optimum cofactor. It displayed catalytic activity maximally at pH 8.0 and 65 °C (as measured over 5 min). The optimum substrate was d-psicose, and the K _m, turnover number (k _cat), and catalytic efficiency (k _cat/K _m) for d-psicose were 227 mM, 32,185 min^?1, and 141 min^?1 mM^?1, respectively. At pH 8.0 and 55 °C, 120 g d-psicose l^?1 was produced from 500 g d-fructose l^?1 after 5 h.     

Translocation,remineralization, and turnover of nitrogen in the roots and rhizomes of Spartina alterniflora (Gramineae)

We used ¹⁵N to quantify rates of N translocation from aerial to belowground tissues, foliar leaching, and turnover and production of root and rhizome biomass in the plant-sediment system of short Spartina alterniflora areas of Great Sippewissett Marsh, Massachusetts. Decay of belowground tissues in litterbag incubations at 1- and 10-cm depths resulted in 80% remineralization of the original plant (¹⁵N-labeled) N and 20% burial after 3 years. Translocation of ¹⁵N from plant shoots in hydrologically controlled laboratory lysimeters maintained under field conditions was 38% of the aboveground pool while leaching of N was 10% from June to October. Most of the translocated N was not retranslocated to new aboveground growth in December but appeared to be either remineralized or buried in the sediment. Injection of ¹⁵N into field stands of grass showed initially high incorporation into plants followed by a continuous decline over the next 7 years yielding a gross tumover time of 1.5–1.6yr. Correcting the gross N turnover for recycling of label via translocation and uptake of remineralized label during this period, a net root and rhizome turnover time of 1.0–1.1 yr was obtained. Combining the turnover time with independent estimates of seasonal belowground biomass yielded an estimate of belowground production of 929–1,022 g C m⁻² yr⁻¹, similar to measurements by traditional biomass harvest, CO² based budgets and models for comparable areas of this marsh. Integration of the production and nitrogen balance estimates for short Spartina marsh yielded translocation, 1.4 g N m⁻² yr⁻¹, leaching, 0.4 g N m⁻² yr⁻¹, remineralization, 14.9–16.3 g N m⁻² yr⁻¹, and burial, 3.7–4.1 g N m⁻² yr⁻¹.     

A d-psicose 3-epimerase with neutral pH optimum from Clostridium bolteae for d-psicose production: cloning, expression, purification, and characterization

d-Tagatose 3-epimerase family enzymes can efficiently catalyze the epimerization of free keto-sugars, which could be used for d-psicose production from d-fructose. In previous studies, all optimum pH values of these enzymes were found to be alkaline. In this study, a d-psicose 3-epimerase (DPEase) with neutral pH optimum from Clostridium bolteae (ATCC BAA-613) was identified and characterized. The gene encoding the recombinant DPEase was cloned and expressed in Escherichia coli. In order to characterize the catalytic properties, the recombinant DPEase was purified to electrophoretic homogeneity using nickel-affinity chromatography. Ethylenediaminetetraacetic acid was shown to inhibit the enzyme activity completely; therefore, the enzyme was identified as a metalloprotein that exhibited the highest activity in the presence of Co²⁺. Although the DPEase demonstrated the most activity at a pH ranging from 6.5 to 7.5, it exhibited optimal activity at pH 7.0. The optimal temperature for the recombinant DPEase was 55 °C, and the half-life was 156 min at 55 °C. Using d-psicose as the substrate, the apparent K _m, k _cat, and catalytic efficiency (k _cat/K _m) were 27.4 mM, 49 s^?1, and 1.78 s^?1 mM^?1, respectively. Under the optimal conditions, the equilibrium ratio of d-fructose to d-psicose was 69:31. For high production of d-psicose, 216 g/L d-psicose could be produced with 28.8 % turnover yield at pH 6.5 and 55 °C. The recombinant DPEase exhibited weak-acid stability and thermostability and had a high affinity and turnover for the substrate d-fructose, indicating that the enzyme was a potential d-psicose producer for industrial production.     

Net primary production and net ecosystem production of a boreal black spruce wildfire chronosequence

Net primary production (NPP) was measured in seven black spruce (Picea mariana (Mill.) BSP)‐dominated sites comprising a boreal forest chronosequence near Thompson, Man., Canada. The sites burned between 1998 and 1850, and each contained separate well‐ and poorly drained stands. All components of NPP were measured, most for 3 consecutive years. Total NPP was low (50–100 g C m^?2 yr^?1) immediately after fire, highest 12–20 years after fire (332 and 521 g C m^?2 yr^?1 in the dry and wet stands, respectively) but 50% lower than this in the oldest stands. Tree NPP was highest 37 years after fire but 16–39% lower in older stands, and was dominated by deciduous seedlings in the young stands and by black spruce trees (>85%) in the older stands. The chronosequence was unreplicated but these results were consistent with 14 secondary sites sampled across the landscape. Bryophytes comprised a large percentage of aboveground NPP in the poorly drained stands, while belowground NPP was 0–40% of total NPP. Interannual NPP variability was greater in the youngest stands, the poorly drained stands, and for understory and detritus production. Net ecosystem production (NEP), calculated using heterotrophic soil and woody debris respiration data from previous studies in this chronosequence, implied that the youngest stands were moderate C sources (roughly, 100 g C m^?2 yr^?1), the middle‐aged stands relatively strong sinks (100–300 g C m^?2 yr^?1), and the oldest stands about neutral with respect to the atmosphere. The ecosystem approach employed in this study provided realistic estimates of chronosequence NPP and NEP, demonstrated the profound impact of wildfire on forest–atmosphere C exchange, and emphasized the need to account for soil drainage, bryophyte production, and species succession when modeling boreal forest C fluxes.     

ROOT PRODUCTION IN FOUR COMMUNITIES IN THE GREAT DISMAL SWAMP

A sequential coring approach was used to measure root biomass and production over 1 year in four different communities within the Great Dismal Swamp. A second method, an implanted bag technique, was also used to measure root production, and values were generally lower using this technique. On all sites, fine roots were the most dynamic root component. Both biomass (1,887 g/m²) and production (354–989 g m ² yr^-1) were highest on the mixed hardwood site, the least flooded site, and second highest on the cedar site, the site with the longest duration of soil saturation (1,033 g/m² and 274–366 g m^-2 yr^-1). The maple-gum (696 g/m² and 59–91 g m^-2 yr^-1) and cypress (824 g/m² and 68–308 g m^-2 yr^-1) sites had similarly low amounts of biomass and rates of production. Environmental parameters that influenced production include frequency and duration of flooding, and soil type. Peaks in belowground production were observed on the most productive sites (mixed hardwood and cedar) in summer and late fall-winter; the other two sites exhibited little seasonal variability. The least flooded stand appears to allocate a greater percentage of net primary production belowground than the more extensively flooded stands. The ratio of above- and belowground allocation appears to change in response to a flooding gradient. This has major implications for ecosystem functions as carbon allocation patterns determine the array of litter types generated (leaves vs. roots) which affect decomposition rates and nutrient availability.     

d-Tagatose production in the presence of borate by resting Lactococcus lactis cells harboring Bifidobacterium longum l-arabinose isomerase

Bifidobacterium longum NRRL B-41409 l-arabinose isomerase (l-AI) was overexpressed in Lactococcus lactis using a phosphate depletion inducible expression system. The resting L. lactis cells harboring the B. longum l-AI were used for production of d-tagatose from d-galactose in the presence of borate buffer. Multivariable analysis suggested that high pH, temperature and borate concentration favoured the conversion of d-galactose to d-tagatose. Almost quantitative conversion (92 %) was achieved at 20 g L^?1 substrate and at 37.5 °C after 5 days. The d-tagatose production rate of 185 g L^?1 day^?1 was obtained at 300 g L^?1 galactose, at 1.15 M borate, and at 41 °C during 10 days when the production medium was changed every 24 h. There was no significant loss in productivity during ten sequential 24 h batches. The initial d-tagatose production rate was 290 g L^?1 day^?1 under these conditions.     

Land use history and resource utilisation from a.d. 400 to the present, at Chibuene, southern Mozambique

This paper discusses changing patterns of resource utilisation over time in the locality of Chibuene, Vilankulos, situated on the coastal plain of southern Mozambique. The macroscopic charcoal, bone and shell assemblages from archaeological excavations are presented and discussed against the off-site palaeoecological records from pollen, fungal spores and microscopic charcoal. The Chibuene landscape has experienced four phases of land use and resource utilisation that have interacted with changes in the environment. Phase 1 (a.d. 400–900), forest savanna mosaic, low intensity cattle herding and cultivation, trade of resources for domestic use. Phase 2 (a.d. 900–1400), forest savanna mosaic, high intensity/extensive cultivation and cattle herding. Phase 3 (a.d. 1400–1800), savanna woodland and progressive decrease in forests owing to droughts. Decline of agricultural activities and higher reliance on marine resources. Possible trade of resources with the interior. Phase 4 (a.d. 1800–1900), open savanna with few forest patches. Warfare and social unrest. Collapse of trade with the interior. Decline in marine resources and wildlife. Loss of cattle herds. Expansion of agriculture locally and introduction of New World crops and clearing of Brachystegia trees. The study shows the importance of combining different environmental resources for elucidating how land use and natural variability have changed over time.     

Characterization of a recombinant l-rhamnose isomerase from Dictyoglomus turgidum and its application for l-rhamnulose production

A putative recombinant enzyme from Dictyoglomus turgidum was characterized and immobilized on Duolite A568 beads. The native enzyme was a 46 kDa tetramer. Its activity was highest for l-rhamnose, indicating that it is an l-rhamnose isomerase. The maximum activities of both the free and immobilized enzymes for l-rhamnose isomerization were at pH 8.0 and 75 °C in the presence of Mn²⁺. Under these conditions, the half-lives of the free and immobilized enzymes were 28 and 112 h, respectively. In a packed-bed bioreactor, the immobilized enzyme produced an average of 130 g l-rhamnulose l^?1 from 300 g l-rhamnose l^?1 after 240 h at pH 8.0, 70 °C, and 0.6 h^?1, with a productivity of 78 g l^?1 h^?1 and a conversion yield of 43 %. To the best of our knowledge, this is the first report describing the enzymatic production of l-rhamnulose.     

Blood-brain barrier transport ofL-pipecolic acid in various rat brain regions

Blood-brain barrier transport ofL-[l-¹⁴C]pipecolic acid was studied in the rat by single intracarotid injection using³H₂O as a diffusible internal standard. Brain uptake index (BUI) forL-[¹⁴C]pipecolic acid (0.036 mM) was found to be 18.1, 10.5, and 12.6 for the cerebral cortex, brain stem, and cerebellum, respectively which was substantially higher than that reported for its analogL-proline in the whole brain. Influx ofL-pipecolic acid into the brain was concentration dependent and differed significantly between the cerebral cortex and the brain stem, and between the cerebral cortex and the cerebellum, but not between the brain stem and the cerebellum. Kinetic study ofL-pipecolic acid influx revealed a low- and a high-capacity uptake mechanisms. The low-capacity saturable component hasK _m values ranging from 38 to 73 μM, andV _max values ranging from 10 to 13 nmol/g/min for the three brain regions. The nonsaturable component has aK _m of 4 mM, aV _max of 200 nmol/g/min and similar diffusion constant (K _d) (0.03 to 0.06 mlg^?1 min^?1) for all three brain regions. A possible role of the two-component brain uptake mechanism in the regulation of the neuronal function ofL-pipecolic acid was suggested.     

Characterization of a recombinant l-fucose isomerase from Caldicellulosiruptor saccharolyticus that isomerizes l-fucose, d-arabinose, d-altrose, and l-galactose

A recombinant l-fucose isomerase from Caldicellulosiruptor saccharolyticus was purified as a single 68 kDa band with an activity of 76 U mg^?1. The molecular mass of the native enzyme was 204 kDa as a trimer. The maximum activity for l-fucose isomerization was at pH 7 and 75°C in the presence of 1 mM Mn²⁺. Its half-life at 70°C was 6.1 h. For aldose substrates, the enzyme displayed activity in decreasing order for l-fucose, with a k _cat of 11,910 min^?1 and a K _m of 140 mM, d-arabinose, d-altrose, and l-galactose. These aldoses were converted to the ketoses l-fuculose, d-ribulose, d-psicose, and l-tagatose, respectively, with 24, 24, 85, 55% conversion yields after 3 h.