Hemipristicola gunterae gen. n., sp. n. (Cestoda: Tetraphyllidea: Phyllobothriidae) from the snaggletooth shark, Hemipristis elongata (Carcharhiniformes: Hemigaleidae), from Moreton Bay, Australia

Helminthological examination of the snaggletooth shark, Hemipristis elongata (Klunzinger) (Carcharhiniformes: Hemigaleidae), from Moreton Bay, Queensland, Australia, yielded a phyllobothriid genus and species previously unknown to science. Hemipristicola gunterae gen. n., sp. n. is described here, and is placed in the subfamily Phyllobothriinae Braun, 1900. Of the other phyllobothriid genera, the new genus most closely resembles Paraorygmatobothrium in that both genera possess bothridia with a single loculus and apical sucker, post-vaginal testes and lateral vitellarium. Hemipristicola, however, differs from Paraorygmatobothrium in the morphology of the proximal bothridial surface microthrix, possessing serrate gladiate spinitriches with marginal serrations restricted to the distal half of the blade, and in the possession of a more extensive uterus, extending anteriorly from the anterior margin of the ovary to well past the level of the cirrus-sac. The new genus also differs from Paraorygmatobothrium by possessing testes that are more than one layer deep. Hemipristicola gunterae further differs from Paraorygmatobothrium species found in hemigaleid sharks in possessing vitelline follicles arranged in two lateral bands that are restricted to the lateral margins of the proglottid and not possessing a cephalic peduncle. Bayesian inference analysis of partial 28S rDNA data shows that H. gunterae forms a sister taxon to species of Paraorygmatobothrium. These two genera were resolved with high posterior probability support in the analysis. Hemipristicola gunterae is only the second phyllobothriid species to be described from Hemipristis elongata from Australian waters, and the fourth from the Australian hemigaleids.     

Conservation biology for suites of species: Demographic modeling for Pacific island kingfishers

Conservation practitioners frequently extrapolate data from single-species investigations when managing critically endangered populations. However, few researchers initiate work with the intent of making findings useful to conservation efforts for other species. We presented and explored the concept of conducting conservation-oriented research for suites of geographically separated populations with similar natural histories, resource needs, and extinction threats. An example was provided in the form of an investigation into the population demography of endangered Micronesian kingfishers (Todiramphus cinnamominus). We provided the first demographic parameter estimates for any of the 12 endangered Pacific Todiramphus species, and used results to develop a population projection matrix model for management throughout the insular Pacific. Further, we used the model for elasticity and simulation analyses with demographic values that randomly varied across ranges that might characterize congener populations. Results from elasticity and simulation analyses indicated that changes in breeding adult survival exerted the greatest magnitude of influence on population dynamics. However, changes in nestling survival were more consistently correlated with population dynamics as demographic rates were randomly altered. We concluded that conservation practitioners working with endangered Pacific kingfishers should primarily focus efforts on factors affecting nestling and breeder survival, and secondarily address fledgling juveniles and helpers. Further, we described how the generalized base model might be changed to focus on individual populations and discussed the potential application of multi-species models to other conservation situations.     

Elevation of atmospheric CO2 and N-nutritional status modify nodulation, nodule-carbon supply, and root exudation of Phaseolus vulgaris L.

Increased root exudation and a related stimulation of rhizosphere-microbial growth have been hypothesised as possible explanations for a lower nitrogen- (N-) nutritional status of plants grown under elevated atmospheric CO₂ concentrations, due to enhanced plant-microbial N competition in the rhizosphere. Leguminous plants may be able to counterbalance the enhanced N requirement by increased symbiotic N₂ fixation. Only limited information is available about the factors determining the stimulation of symbiotic N₂ fixation in response to elevated CO₂.In this study, short-term effects of elevated CO₂ on quality and quantity of root exudation, and on carbon supply to the nodules were assessed in Phaseolus vulgaris, grown in soil culture with limited (30 mg N kg⁻¹ soil) and sufficient N supply (200 mg N kg⁻¹ soil), at ambient (400 μmol mol⁻¹) and elevated (800 μmol mol⁻¹) atmospheric CO₂ concentrations.Elevated CO₂ reduced N tissue concentrations in both N treatments, accelerated the expression of N deficiency symptoms in the N-limited variant, but did not affect plant biomass production. ¹⁴CO₂ pulse-chase labelling revealed no indication for a general increase in root exudation with subsequent stimulation of rhizosphere microbial growth, resulting in increased N-competition in the rhizosphere at elevated CO₂. However, a CO₂-induced stimulation in root exudation of sugars and malate as a chemo-attractant for rhizobia was detected in 0.5-1.5 cm apical root zones as potential infection sites. Particularly in nodules, elevated CO₂ increased the accumulation of malate as a major carbon source for the microsymbiont and of malonate with essential functions for nodule development. Nodule number, biomass and the proportion of leghaemoglobin-producing nodules were also enhanced. The release of nod-gene-inducing flavonoids (genistein, daidzein and coumestrol) was stimulated under elevated CO₂, independent of the N supply, and was already detectable at early stages of seedling development at 6 days after sowing.     

Increased salicylate concentrations in urine of human volunteers after consumption of cranberry juice

The aim of this study was to assess whether regular consumption of cranberry juice results in elevations in urinary salicylate concentrations in persons not taking salicylate drugs. Two groups of healthy female subjects (11/group) matched for age, weight, and height consumed 250 mL of either cranberry juice or a placebo solution three times a day (i.e., 750 mL/day) for 2 weeks. At weekly intervals, salicylic acid and salicyluric acid (the major urinary metabolite of salicylic acid) concentrations were determined in urine by HPLC with electrochemical detection. Concentrations of salicylic acid in plasma were also determined. Consumption of cranberry juice was associated with a marked increase (p < 0.001) of salicyluric and salicylic acids in urine within 1 week of the intervention. After 2 weeks, there was also a small but significant (p < 0.05) increase in salicylic acid in plasma. The regular consumption of cranberry juice results in the increased absorption of salicylic acid, an anti-inflammatory compound that may benefit health.     

Hydrogen peroxide induced oxidative stress damage and antioxidant enzyme response in Caco-2 human colon cells

Studies were conducted to evaluate the cell damage caused by exposing human colon carcinoma cells, Caco-2, to hydrogen peroxide at concentrations varying from 0 to 250 microM for 30 min. Evaluation of cell viability, as measured by trypan blue dye exclusion test, showed that the loss of viability was < 5% at concentrations up to 250 microM hydrogen peroxide. Cell membrane damage and DNA damage as measured by the leakage of lactate dehydrogenase and the comet assay, respectively, were significantly high at concentrations >100 microM hydrogen peroxide compared to those of the control. Antioxidant mechanisms in Caco-2 cells were evaluated by measuring catalase, superoxide dismutase, and glutathione peroxidase activities. Catalase activities remained constant in cells treated with 50-250 microM hydrogen peroxide. Superoxide dismutase activity decreased, whereas glutathione peroxidase activity increased in cells treated with H(2)O(2) concentrations of >50 microM. This study showed that with increasing hydrogen peroxide concentration, cell membrane leakage and DNA damage increased, whereas the three antioxidant enzymes responded differently, as shown by mathematical models.     

Organic acids influence iron uptake in the human epithelial cell line Caco-2

It has previously been suggested that organic acids enhance iron absorption. We have studied the effect of nine organic acids on the absorption of Fe(II) and Fe(III) in the human epithelial cell line Caco-2. The effect obtained was dose-dependent, and the greatest increase (43-fold) was observed for tartaric acid (4 mmol/L) on Fe(III) (10 micromol/L). Tartaric, malic, succinic, and fumaric acids enhanced Fe(II) and Fe(III) uptake. Citric and oxalic acid, on the other hand, inhibited Fe(II) uptake but enhanced Fe(III) uptake. Propionic and acetic acid increased the Fe(II) uptake, but had no effect on Fe(III) uptake. Our results show a correlation between absorption pattern and chemical structure; e.g. hydroxyl groups, in addition to carboxyls, were connected with a positive influence. The results may be important for elucidating factors affecting iron bioavailability in the small intestine and for the development of foods with improved iron bioavailability.     

Discriminating between the six isomers of dicaffeoylquinic acid by LC-MS(n)

The fragmentation behavior of all six dicaffeoylquinic acids (diCQA) has been investigated using LC-MS(4). It is possible to discriminate between each of the isomers including those for which commercial standards are not available. For diCQA, the ease of removal of the caffeoyl residue during fragmentation is 1 approximately = 5 > 3 > 4. The distinctive fragmentation observed for the little-studied 1,4-dicaffeoylquinic acid involves elimination of the C1 caffeoyl residue, repeated dehydrations leading to the aromatization of the quinic acid moiety, and its decarboxylation. It is suggested that this process is initiated by the C1 carboxyl protonating the C5 hydroxyl in the inverted chair conformer, followed by its protonating the C1 caffeoyl residue in the favored chair conformation. The fragmentation of 1-caffeoylquinic acid is indistinguishable from that of the commercially available 5-caffeoylquinic acid, but these two isomers can be distinguished easily by their facile chromatographic resolution on reversed phase packings. The hierarchical key previously developed for characterizing chlorogenic acids has been extended to accommodate 1-caffeoylquinic acid and the 1-acyl dicaffeoylquinic acids.     

Effects of a plant parasitic nematode (Heterodera trifolii) on clover roots and soil microbial communities

We studied the effects of the root endoparasitic nematode Heterodera trifolii on rhizodeposition and the root architecture of white clover (Trifolium repens). Rhizosphere solutions were collected from the root systems of plants growing with and without H. trifolii (200 juveniles per inoculated plant) in sand-based microlysimeters. The organic carbon (C) content of these solutions was analyzed, and they were applied to plant-free soils to investigate microbial responses. Although plant biomass was unaffected by nematodes, the architecture of the root systems was significantly altered, with a decrease in overall root length and an increase in the density of lateral branches from the primary root. The presence of nematodes reduced the concentration of organic compounds in the rhizosphere solutions but only on the final sampling date (75 days). Analysis of microbial signature phospholipid fatty acids revealed no change in the structure of the microbial communities in soils to which rhizosphere solutions were applied. However, these microorganisms did respond with changes in substrate utilization patterns (community-level physiological profiles). Microbes in soils that received rhizosphere solutions from the nematode-infected clover showed lower utilization of most substrates but higher utilization of oligosugars. These responses appear to be related to changes in roots and rhizodeposition associated with nematode infection of clover roots. The results of this study suggest that root herbivory can negatively impact carbon-limited soil microbial communities via changes in root architecture that moderate rhizodeposition.     

Hierarchical scheme for LC-MSn identification of chlorogenic acids

The fragmentation behavior of 18 chlorogenic acids that are not substituted at position 1 has been investigated using LC-MS(4) applied to a methanolic coffee bean extract and commercial cider (hard cider). Using LC-MS(3), it is possible to discriminate between each of the three isomers of p-coumaroylquinic acid, caffeoylquinic acid, feruloylquinic acid, and dicaffeoylquinic acid, and a hierarchical key has been prepared to facilitate this process when standards are not available. MS(4) fragmentations further support these assignments, but were not essential in reaching them. The distinctive behavior of 4-acyl and 3-acyl chlorogenic acids compared with the 5-acyl chlorogenic acids is a key factor permitting these assignments. The fragmentation patterns are dependent upon the particular stereochemical relationships between the individual substituents on the quinic acid moiety. Fragmentation is facilitated by 1,2-acyl participation and proceeds through quinic acid conformers in which the relevant substituents transiently adopt a 1,3-syn-diaxial relationship. Selected ion monitoring at m/z 529 clearly indicated the presence in coffee of six caffeoylferuloylquinic acid isomers, whereas previously only two or three had been demonstrated. The hierarchical key permitted specific structures to be assigned to each of the six isomers. These assignments are internally consistent and consistent with the limited data previously available.