Characterization of intracellular and extracellular saxitoxin levels in both field and cultured Alexandrium spp. samples from Sequim Bay, Washington

Traditionally, harmful algal bloom studies have primarily focused on quantifying toxin levels contained within the phytoplankton cells of interest. In the case of paralytic shellfish poisoning toxins (PSTs), intracellular toxin levels and the effects of dietary consumption of toxic cells by planktivores have been well documented. However, little information is available regarding the levels of extracellular PSTs that may leak or be released into seawater from toxic cells during blooms. In order to fully evaluate the risks of harmful algal bloom toxins in the marine food web, it is necessary to understand all potential routes of exposure. In the present study, extracellular and intracellular PST levels were measured in field seawater samples (collected weekly from June to October 2004–2007) and in Alexandrium spp. culture samples isolated from Sequim Bay, Washington. Measurable levels of intra- and extra-cellular toxins were detected in both field and culture samples via receptor binding assay (RBA) and an enzyme-linked immunosorbent assay (ELISA). Characterization of the PST toxin profile in the Sequim Bay isolates by pre-column oxidation and HPLC-fluorescence detection revealed that gonyautoxin 1 and 4 made up 65 ± 9.7 % of the total PSTs present. Collectively, these data confirm that extracellular PSTs are present during blooms of Alexandrium spp. in the Sequim Bay region.     

Cadmium uptake by Lupinus albus (L.): Cadmium excretion,a possible mechanism of cadmium tolerance

Cadmium (Cd) uptake by white lupin (Lupinus albus) was studied at low Cd concentrations (0.05nM to 5 μM) in hydroponic solution. Ten 12‐day old seedlings were pretreated in 0.5 mM CaCl₂ solution in presence and absence of metabolic inhibitors (DCCD, DNP or NaN3). Cadmium solutions were labelled with carrier free ¹⁰⁹CdCl2. Cadmium uptake was measured after a 2 h desorption in unlabelled CdCl2 solution. In the absence of any metabolic inhibitor and at 5 [μM Cd, roots absorbed 235.23 μg Cd/g root dry weight. Over the range of lnM to 5 μM Cd, exchangeable Cd represented approximately 5% of the absorbed fraction, and about 25 % of the total absorbed Cd was adsorbed to the root. Cadmium was passively absorbed to about 30% as observed in the presence of the inhibitor (DCCD). Ative absorption which represented 70% of Cd uptake involved H⁺‐ATPase carriers. Cadmium absorption was reduced to 30 and 20% in presence of lanthanum (La³⁺) and zinc (Zn²⁺), respectively which suggested that calcium (Ca), Cd, and Zn use the same carriers. Cadmium uptake in presence of DNP or NaN3 was approximately 4‐ fold that in control. Data showed presomption for an excretion of Cd out of root cells which could be the expression of a detoxification process limiting cell contamination.     

Host/parasite relationship in the in vitro infection of rat gliocytes by Neospora caninum: evaluation of cell respiration

Neospora caninum is a protozoon that causes abortion in cattle and neuromuscular lesions in dogs, with the formation of cysts mainly in the central nervous system. Since N. caninum is an intracellular parasite with tropism for the cells of nervous system, this study evaluated the respiratory metabolism of glial cells infected by this* parasite. Glial cultures obtained from the cerebral cortex of newborn rats were kept in DMEM enriched with 10% fetal bovine serum, 1 mM pyruvic acid and 2 mM of L-glutamine. They were infected at a ratio of approximately 1:1 (cell/parasite). Oxygen consumption was evaluated by polarography in the non infected and N. caninum infected groups, 24 and 72 h following infection. Glial cell respiration after 24 and 72 h was 307.2 +/- 34.7 and 308.9 +/- 64.1 microL of oxygen per mug of total protein per minute, and 566.2 +/- 54.6 and 579 +/- 117.5 microL O2/microg of total protein/minute in the control and infected groups, respectively. These results show that N. caninum does not interfere with glial respiration in vitro.     

Variation of Phyllosphere Microflora of Different Rice Varieties in Sri Lanka and its Relationship to Leaf Anatomical and Physiological Characters

Anatomical and physiological characters of the leaf surface and its physico‐chemical environment substantially influence the density and diversity of phyllosphere‐inhabiting microorganisms, which may include natural antagonists of important pathogens. The objective of this investigation was to quantify the phyllosphere (i.e. leaf surface) microbial population in a range of rice varieties grown in Sri Lanka and to identify the leaf anatomical and physiological characters that determine the density and diversity of phyllosphere microbes. Fifteen rice varieties including both traditional and new high‐yielding varieties were used in a planthouse experiment and a field experiment in two consecutive seasons to quantify the phyllosphere microbial population and measure leaf characters that may influence it. There were highly significant intervarietal variations in the density and diversity of epiphytic bacterial, fungal and total microbial populations under both planthouse and field conditions. However, there was no difference between traditional and new, high‐yielding varieties in their capacity to harbour epiphytic microbes in the phyllosphere. Total microbial density (TMD) under both conditions showed positive correlations with leaf hair density, stomatal density and transpiration rate. Under planthouse conditions, TMD was also positively correlated with leaf hair length and negatively correlated with leaf temperature. These correlations can be explained in terms of providing favourable microsites on the phylloplane for epiphytic microbial growth.     

Absorption of calcium fumarate salts is equivalent to other calcium salts when measured in the rat model

Calcium absorption from fumarate salts (calcium fumarate and calcium malate fumarate), which have recently been considered for use as sources for food and beverage enrichment, was compared to that from calcium citrate malate, calcium citrate, and calcium carbonate. Salts were instrinsically labeled with 45Ca and orally administered to Sprague-Dawley rats. Fractional absorption of calcium from each salt was determined using the femur uptake model. Fractional absorption from the five salts (0.30-0.27) was not significantly different (p > 0.05). Thus, when measured in the rat model, calcium from calcium fumarate and calcium malate fumarate is absorbed equally well as compared to other salts, which are common calcium sources in many foods, beverages, and supplements.     

Observation of X-ray lines from a gamma-ray burst (GRB991216): evidence of moving ejecta from the progenitor

We report on the discovery of two emission features observed in the x-ray spectrum of the afterglow of the gamma-ray burst (GRB) of 16 December 1999 by the Chandra X-ray Observatory. These features are identified with the Ly(alpha) line and the narrow recombination continuum by hydrogenic ions of iron at a redshift z = 1.00 +/- 0.02, providing an unambiguous measurement of the distance of a GRB. Line width and intensity imply that the progenitor of the GRB was a massive star system that ejected, before the GRB event, a quantity of iron approximately 0.01 of the mass of the sun at a velocity approximately 0.1 of the speed of light, probably by a supernova explosion.     

Determination of angiotensin I-converting enzyme activity in equine blood: lack of agreement between methods of analysis

Angiotensin-I converting enzyme (ACE) is a key regulator of blood pressure, electrolytes and fluid homeostasis through conversion of angiotensin I into angiotensin II. Recently, a genetic polymorphism of the ACE gene, which accounts for 47% of the variation of ACE activity in blood, has been advocated as a biomarker of athletic aptitude. Different methods of analysis and determination of ACE activity in plasma have been used in human and equine research without a consensus of a "gold standard" method. Different methods have often been used interchangeably or cited as being comparable in the existing literature; however, the actual agreement between assays has not been investigated. Therefore, in this study, we evaluated the level of agreement between three different assays using equine plasma obtained from 29 horses. Two spectrophotometric assays using Furylacryloyl-phenylalanyl-glycyl-glycine as substrate and one fluorimetric assay utilizing o-aminobenzoic acid-FRK-(Dnp)P-OH were employed. The results revealed that the measurements from the different assays were not in agreement, indicating that the methods should not be used interchangeably for measurement of equine ACE activity. Rather, a single method of analysis should be adopted to achieve comparable results and critical appraisal of the literature is needed when attempting to compare results obtained from different assays.