Prostaglandin E(2) and I(2) facilitate noxious heat-induced spike discharge but not iCGRP release from rat cutaneous nociceptors

The bradykinin-induced sensitization of cutaneous nociceptors to heat was previously shown to be abolished by cyclooxygenase blockade suggesting that endogenous prostaglandins exerted a heat-sensitizing action. The present study aimed at investigating the effects of exogenous prostaglandin E(2) (PGE(2)) and I(2) (PGI(2)) on noxious heat-evoked responses of rat cutaneous nociceptors. As neuropeptides including calcitonin gene-related peptide (CGRP) can be released from the peptidergic subset of heat-sensitive nociceptors, both the spike-generating (afferent) and CGRP-releasing (efferent) responses to heat stimulation were assessed by recording action potentials from single cutaneous C-fibers and measuring immunoreactive CGRP (iCGRP) release from isolated skin flaps, respectively. A combination of PGE(2) and PGI(2) (100 microM for both) unlike 10 microM PGE(2) or PGI(2) increased the number of spikes discharged during a noxious heat stimulus whereas the heat threshold remained unchanged. In contrast, 100 microM PGE(2) plus PGI(2) failed to increase the iCGRP release induced by noxious heat (47 degrees C) from the isolated rat skin. PGE(2) (100 microM), however, augmented the iCGRP-releasing effect of protons (pH 5.7). The adenylyl cyclase activator forskolin and the protein kinase C activator phorbol ester (PMA, 10 microM for both) facilitated heat-induced iCGRP release whereas increasing the intracellular Ca(2+) concentration by 10 microM ionomycin produced a desensitization of the response. In conclusion, PGE(2) plus PGI(2) can sensitize the afferent function of nociceptors in the rat skin, by increasing heat-induced spike discharge, but not the heat-induced efferent response i.e. iCGRP release. This discrepancy might reflect the differences between mechanisms of Na(+) channel-dependent spike generation and Ca(2+)-dependent neuropeptide release.     

The Stream Inlet to a Shallow Bay of a Drinking Water Reservoir,a ‘Hot‐Spot’ for Microcystis Blooms Initiation

Stream inlets into shallow bays of reservoirs and lakes may be ‘hot‐spots’ for toxic cyanobacterial bloom initiation. These ‘hot‐spots’ may be connected with the permanent inflow of high nutrient concentrations from the catchment, optimal physical conditions (wind protected areas) that occur in shallow areas and/or ineffective top‐down control. Four sampling sites along a transect from stream to reservoir in a shallow bay of Sulejow Reservoir (Poland) were studied to test the above hypothesis, comprising a transition zone between lotic and pelagic habitats. Investigations showed that stream inlet into shallow bay acted as incubator for Microcystis blooms. The nutrient level, especially phosphorus, was identified as the major cause of cyanobacterial bloom growth. The increase of Microcystis biomass strongly correlated with increasing microcystin concentrations, however, a relationship with microcystin content was not observed. Toxicity of bloom demonstrated seasonal variability, reaching its maximum at the initial phase of bloom. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Structure and physicochemical properties of barley non-starch polysaccharides — I. Water-extractable β-glucans and arabinoxylans

Water-soluble non-starch polysaccharides were extracted from a Canadian malting barley (cv. Harrington) by sequential treatment with water at 40 °C (WE40) and 65 °C (WE65). The yields were 1.4 and 1.3% (w/w), respectively, of the dry barley grist. The WE40 extract was composed of 82.5% glucose, 8.9% xylose, and 7.0% arabiose residues, whereas WE65 contained 93.3% Glc, 3.3% Xyl, and 2.5% Ara. Only minute amounts of mannose and galactose residues were found in either fraction. Both extracts were further fractionated by stepwise (NH₄)₂SO₄ precipitation into several polysaccharide populations. Subfractions from both extracts, obtained up to 45% saturation with (NH₄)₂SO₄, contained mostly β-glucans, whereas subfractions precipitated at increasing saturation levels of (NH₄)₂SO₄ (45–100%) contained progressively more arabinoxylans and less β-glucans. Compared to WE40, the WE65 extract was enriched in β-glucan populations with higher molecular size, higher limiting viscosity values, and higher content of β-(1 → 4) linkages. The ratio of tri-/ tetrasaccharide oligomers was also higher in β-glucans extracted at 65 °C than those extracted at 40 °C. Arabinoxylans in both extracts, WE40 and WE65, were highly substituted and contained large proportions of doubly substituted xylose residues.     

Long-Term Patterns in the Population Dynamics of Daphnia longispina,Leptodora kindtii and Cyanobacteria in a Shallow Reservoir: A Self-Organising Map (SOM) Approach

The recognition of long-term patterns in the seasonal dynamics of Daphnia longispina, Leptodora kindtii and cyanobacteria is dependent upon their interactions, the water temperature and the hydrological conditions, which were all investigated between 1999 and 2008 in the lowland Sulejow Reservoir. The biomass of cyanobacteria, densities of D. longispina and L. kindtii, concentration of chlorophyll a and water temperature were assessed weekly from April to October at three sampling stations along the longitudinal reservoir axis. The retention time was calculated using data on the actual water inflow and reservoir volume. A self-organising map (SOM) was used due to high interannual variability in the studied parameters and their often non-linear relationships. Classification of the SOM output neurons into three clusters that grouped the sampling terms with similar biotic states allowed identification of the crucial abiotic factors responsible for the seasonal sequence of events: cluster CL-ExSp (extreme/spring) corresponded to hydrologically unstable cold periods (mostly spring) with extreme values and highly variable abiotic factors, which made abiotic control of the biota dominant; cluster CL-StSm (stable/summer) was associated with ordinary late spring and summer and was characterised by stable non-extreme abiotic conditions, which made biotic interactions more important; and the cluster CL-ExSm (extreme/summer), was associated with late spring/summer and characterised by thermal or hydrological extremes, which weakened the role of biotic factors. The significance of the differences between the SOM sub-clusters was verified by Kruskal-Wallis and post-hoc Dunn tests. The importance of the temperature and hydrological regimes as the key plankton-regulating factors in the dam reservoir, as shown by the SOM, was confirmed by the results of canonical correlation analyses (CCA) of each cluster. The demonstrated significance of hydrology in seasonal plankton dynamics complements the widely accepted pattern proposed by the plankton succession model for lakes, the PEG (Plankton Ecology Group), and may be useful for the formulation of management decisions in dam reservoirs.     

Role of Environmental Factors and Toxic Genotypes in the Regulation of Microcystins-Producing Cyanobacterial Blooms

The aim of this study was to understand: (1) how environmental conditions can contribute to formation of Microcystis-dominated blooms in lowland, dam reservoirs in temperate climate—with the use of quantitative molecular monitoring, and (2) what is the role of toxic Microcystis genotypes in the bloom functioning. Monitoring of the Sulejow Reservoir in 2009 and 2010 in two sites Tresta (TR) and Bronislawow BR), which have different morphometry, showed that physicochemical conditions were always favorable for cyanobacterial bloom formation. In 2009, the average biomass of cyanobacteria reached 13 mg L^?1 (TR) and 8 mg L^?1 (BR), and in the second year, it decreased to approximately 1 mg L^?1 (TR and BR). In turns, the mean number of toxic Microcystis genotypes in the total Microcystis reached 1 % in 2009, both in TR and BR, and in 2010, the number increased to 70 % in TR and 14 % in BR. Despite significant differences in the biomass of cyanobacteria in 2009 and 2010, the mean microcystins (MCs) concentration and toxicity stayed at a similar level of approximately 1 μg L^?1. Statistical analysis indicated that water retention time was a factor that provided a significant difference between the two monitoring seasons and was considered a driver of the changes occurring in the Sulejow Reservoir. Hydrologic differences, which occurred between two studied years due to heavy flooding in Poland in 2010, influenced the decrease in number of Microcystis biomass by causing water disturbances and by lowering water temperature. Statistical analysis showed that Microcystis aeruginosa biomass and 16S rRNA gene copy number representing Microcystis genotypes in both years of monitoring could be predicted on the basis of total and dissolved phosphorus concentrations and water temperature. In present study, the number of mcyA gene copies representing toxic Microcystis genotypes could be predicted based on the biomass of M. aeruginosa. Moreover, MCs toxicity and concentration could be predicted on the basic of mcyA gene copy number and M. aeruginosa (biomass, 16S rRNA), respectively. Present findings may indicate that Microcystis can regulate the number of toxic genotypes, and in this way adjust the whole bloom to be able to produce MCs at the level which is necessary for its maintenance in the Sulejow Reservoir under stressful hydrological conditions.     

Horizontal migration of zooplankton in a littoral zone of the lowland Sulejow Reservoir (Central Poland)

Horizontal migrations of zooplankton between macrophyte patches and open areas were investigated in the sparsely vegetated littoral zone of the Sulejow Reservoir in June-July 2000 and 2001, using one-litre plastic traps. Large-bodied zooplankton: daphnids and copepods generally swam towards the open water at dusk and towards submerged macrophytes at dawn. Small-bodied zooplankton (Bosmina sp., Chydorus sp.) did not show any pattern of horizontal movement. At the time of the research the phytoplankton community was dominated by eatable diatoms (Cyclotella sp.), whose biomass reached 14 mg l⁻¹. Thus, bottom-up forces (food scarcity) are not likely to be responsible for the observed zooplankton migrations. Analyses of fish stomach contents showed high contribution of large zooplankters to the food of juvenile roach (Rutilus rutilus) and perch (Perca fluviatilis) which densely inhabited the littoral zone of reservoir. High fish pressure in the littoral zone along with high density of the predatory cladoceran, Leptodora kindtii in the open water, suggest that top-down forces (predatory pressure) were responsible for the migration of large zooplankton. At dusk predatory pressure of fish fry exceeded that of L. kindtii, forcing endangered zooplankton to escape from macrophytes towards open water. The opposite situation occurred at dawn. The consequences of the relationships for both zooplankton and fish fry communities dynamics are discussed.

     

Structure and physicochemical properties of barley non-starch polysaccharides—II. Alkaliextractable β-glucans and arabinoxylans

Three fractions containing hemicellulosic material were obtained by sequential extraction of barley residue (left after removal of water-extractable polysaccharides) with saturated barium hydroxide [Ba(OH)₂ fraction], distilled water [Ba(OH)₂/H₂O fraction], and 1 m sodium hydroxide [NaOH fraction]. The yields of the fractions were 1.6, 1.7, and 2.6% (w/w), respectively, of the dry barley grist. The Ba(OH)₂ fraction contained mainly arabinose and xylose, 35.8% and 60.9%, respectively. The Ba(OH)₂/H₂O fraction in addition to 26.7% Ara and 36.6% Xyl contained also 34.8% Glc. The NaOH fraction was composed of 14.2% Ara, 44.0% Xyl, and 40.9% Glc. The Ba(OH)₂/H₂O and NaOH extracts were further fractionated by stepwise (NH₄)₂SO₄ precipitation into several subfractions with varying amounts of β-glucans and arabinoxylans. β-Glucans in Ba(OH)₂/H₂O and NaOH fractions were characterized by high ratios of β-(1→4)/β-(1→3) linkages, large amounts of contiguously linked β-(1→4) segments, and high ratios of cellotriosyl/cellotetraosyl units. The alkali-extractable arabinoxylans, especially those NaOH-extractable, were characterized by a very low degree of substitution, high xylose/arabinose ratio, and a small content of doubly substituted xylose residues. Some populations of arabinoxylans displayed structural features that would enable them to self-associate or to interact with β-glucans.     

Sequential solvent extraction and structural characterization of polysaccharides from the endosperm cell walls of barley grown in different environments

The objective of this study was to examine the composition and molecular structure of the endosperm cell walls (CW) derived from barley grain grown in three environments in Canada, and differing in grain hardness, protein, and total β-glucan contents. The endosperm CW were isolated from barley, cv. Metcalfe, grown in Davidson, SK (Sample A), Hythe, AB (sample B), and Hamiota, MB (sample C). The CW were sequentially extracted with water at 65 ^oC, saturated Ba(OH)₂, again with water at 25 ^oC, and 1 M NaOH, resulting in fractions designated WE65, BaE, Ba/WE, and NaE, respectively. The monosaccharide analysis indicated the presence of β-glucans, arabinoxylans, and small amounts of arabinogalactans, glucomannans, and xyloglucans. Cellulose was detected in the CW remnants. The CW of sample A, exhibiting a lower grain hardness than sample B, contained the lowest amount of β-glucans, but the highest amount of arabinoxylans and the mannose-containing polysaccharides. The CW of sample C, characterized by very high protein content in the grain, contained the highest amount of β-glucans and the lowest amount of other polysaccharides. Polysaccharides in the CW of sample B, exhibiting the highest grain hardness, were characterized by the highest weight average molecular weights (M_w). β-Glucans in the CW of Sample B showed the highest ratio of DP3/DP4 and the longest cellulosic fragments in the polymeric chains. Of the three barley samples, arabinoxylans in the endosperm CW of sample A exhibited the lowest degree of branching, the highest amount of unsubstituted Xyl residues, and the highest ratio of singly to doubly substituted Xylp. The highest water solubility of the CW of sample C was associated with the highest concentration of β-glucans, the lowest DP3/DP4 ratio, and the lowest M_w of the polymeric constituents. Arabinoxylans with the lowest amount of doubly substituted but the highest amount of unsubstituted xylose residues and long sequences of unsubstituted xylan regions were found in the NaE fractions. The NaE fractions showed a high ratio of →4)-Glcp-(1→ to →3)-Glcp-(1→ linkages and some →4)-Manp-(1→ linkages, indicating a high level of long cellulosic regions in β-glucan chains and the presence of glucomannans.