PubMed Health⌕ Search

Biomedical subjects

G L Boyer

Publications and source records attributed to G L Boyer.

8 recordsLinked to original sources

Quantification of toxic Microcystis spp. during the 2003 and 2004 blooms in western Lake Erie using quantitative real-time PCR.

In August of 2003 and August of 2004, blooms of potentially toxic cyanobacteria Microcystis spp. persisted in western Lake Erie. Samples collected from the bloom were analyzed for the cyanobacterial toxin microcystin and the presence of Microcystis spp. cells. Estimates of microcystin toxicity exceeding 1 microg L(-1) (microcystin-LR activity equivalents), the safety limit set by the World Health Organization, were found from the samples in both 2003 and 2004. The presence of Microcystis spp. in water samples was confirmed through standard polymerase chain reaction (PCR) using a combination of four primer sets. Quantification of Microcystis was accomplished by a real-time PCR assay utilizing specific primer-Taq-man probe sets targeted on a conserved, Microcystis-specific 16S rDNA fragment and a microcystin toxin synthetase gene mcyD. This approach allowed us to specifically study the distribution and abundance of toxic Microcystis in the lake in contrast to previous studies that have assessed Microcystis populations with less refined methods. On the basis of quantification by quantitative real-time PCR analysis, the total abundance of Microcystis cells in the bloom area varied from 4 x 10(8) to 2 x 10(3) cells L(-1). The results of this study provide novel insight regarding the distribution and abundance of Microcystis spp. in the western basin of Lake Erie, a region plagued in recent years by large-scale (>20 km2) blooms. Our results suggest that the Maumee River and Bay may serve as a source for Microcystis to western and central Lake Erie.

Bacterial Toxins↗

Sterol composition of Aureoumbra lagunensis, the Texas brown tide alga.

Aureoumbra lagunensis, the alga responsible for the "Texas brown tide", contains (E)-24-propylidenecholesterol (35.7% of total sterols) as its dominant sterol, in common with other members of the Pelagophyceae. Other major sterols are stigmasterol (22.2%), sitosterol (19.2%), cholesterol (14.1%), and (24R)-24-propylcholesterol (5.2%). Trace amounts of 24-methylenecholesterol, crinosterol, clerosterol, campesterol, dihydrobrassicasterol, and 24-isopropylcholesterol were also detected.

Chromatography, High Pressure Liquid↗

High performance liquid chromatography coupled with post-column electrochemical oxidation for the detection of PSP toxins.

High Performance Liquid Chromatography (HPLC) is an important tool for the study of PSP toxins. It provides an alternative to bioassays and gives the concentration of individual toxin isomers. The current HPLC protocol uses a post-column chemical reaction system (PCRS) to oxidize the saxitoxin ring system to form a fluorescent chromophore. This oxidation is sensitive to changes in the flow rate, temperature, pH and age of the reagents. We have previously shown that this oxidation can be accomplished using electrochemical techniques. Termed the electrochemical oxidation system (ECOS), this approach provides a simpler alternative to the traditional PCRS-based HPLC system. A detailed description of the construction and maintenance of an HPLC-ECOS system for the analysis of PSP toxins is presented. Comparisons of the mouse bioassay, HPLC-PCRS and HPLC-ECOS system are presented for three different sample matrices: toxic dinoflagellates (Alexandrium tamarense), geoduck (Panopea generosa) and scallops (Placopectin magellanicus). In all three cases, the correlation of the HPLC-ECOS system to the mouse bioassay is similar to that obtained using the HPLC-PCRS system for the analysis of PSP toxins.

Animals↗

Siderophore-Mediated Aluminum Uptake by Bacillus megaterium ATCC 19213.

The bacterium Bacillus megaterium ATCC 19213 is known to produce two hydroxamate siderophores, schizokinen and N-deoxyschizokinen, under iron-limited conditions. In addition to their high affinity for ferric ions, these siderophores chelate aluminum. Aluminum was absorbed by B. megaterium ATCC 19213 through the siderophore transport receptor, providing an extra pathway for aluminum accumulation into iron-deficient bacteria. At low concentrations of the metal, siderophore-mediated uptake was the dominant process for aluminum accumulation. At high concentrations of aluminum, passive transport dominated and siderophore production slowed the passive transport of aluminum into the cell. Siderophore production was affected by the aluminum content in the media. High concentrations of aluminum increased production of siderophores in iron-limited cultures, and this production continued into stationary phase. Aluminum did not stimulate siderophore production in iron-replete cultures. The production of siderophores markedly affected aluminum uptake. This has direct implications on the toxicity of heavy metals under iron-deficient conditions.

Journal Article↗

Isolation and spectral characteristics of four toxins from the dinoflagellate Ptychodiscus brevis.

Four ichthyotoxins were isolated from crude toxic extracts of Ptychodiscus brevis using a combination of solvent partitioning, thin layer chromatography and reversed phase high pressure liquid chromatography. The toxins were analyzed by mass, infrared, 1H- and 13C-NMR spectroscopy and were found to constitute two structural families of two toxins each: brevetoxins 1 and 2 and brevetoxins 3 and 4. Comparison with literature data indicates that brevetoxins 1 and 2 are identical to the previously described and characterized 11-ring polyether toxins brevetoxins C and B, respectively. The other two compounds (brevetoxins 3 and 4) also represent a structural pair (with chloroacetone and alpha-methylene-propanal side chains, respectively) which has a different, but related, basic ring-structure.

Animals↗

Accumulation and transport of abscisic Acid and its metabolites in ricinus and xanthium.

When intact plants of Xanthium strumarium L. were water stressed, the youngest leaves accumulated the highest levels of abscisic acid (ABA). On the other hand, when leaves of different ages were detached and then stressed, the capacity to produce ABA was highest in the mature leaves. Radioactive ABA was transported from mature leaves to the shoot tips and young leaves, as well as to the roots, as evidenced by the presence of radioactive ABA and phaseic acid in the xylem exudate coming from the roots. Thus, ABA was recirculated in the plant, moving down the stem in the phloem and back up in the transpiration stream to the mature leaves. Phloem exudate collected by the use of the EDTA technique had a high concentration of ABA and phaseic acid which increased several-fold after water stress. The high ABA levels in immature leaves and apical buds are, therefore, mainly due to import from older leaves, rather than to in situ synthesis.In Ricinus communis L. cv gibsonii labeled ABA was rapidly exported from a mature leaf as measured by the appearance of radioactive ABA in the phloem collected from incisions in the stem below the treated leaf. After 2 h, small amounts of radioactive phaseic acid and dihydrophaseic acid were also present in the phloem exudate. The glucosyl ester of abscisic acid, beta-d-glucopyranosyl abscisate, was hydrolyzed before entering the phloem and moved as free ABA. It is concluded that the glucosyl ester of ABA itself cannot cross cell membranes and is, therefore, not transported.

Journal Article↗

Isolation and Quantitation of beta-d-Glucopyranosyl Abscisate from Leaves of Xanthium and Spinach.

From previous work (Zeevaart 1980 Plant Physiol 66: 672-678) Xanthium leaves are known to contain a high level of alkali-hydrolyzable conjugated abscisic acid. This abscisic acid conjugate has been isolated and identified by mass spectrometry, nuclear magnetic resonance, and chemical and enzymic degradation techniques, as the glucosyl ester of abscisic acid, beta-d-glucopyranosyl abscisate. The glucosyl ester of abscisic acid was the only abscisic acid conjugate found in Xanthium leaves. It was also isolated from spinach leaves.An insignificant amount of the glucosyl ester of abscisic acid partitioned into diethyl ether, whereas 12% partitioned into ethyl acetate. Consequently, removal of abscisic acid by partitioning with ethyl acetate will result in considerable losses of the glucosyl ester of abscisic acid from the aqueous phase. Diethyl ether is, therefore, recommended for separation of abscisic acid and the glucosyl ester of abscisic acid by solvent partitioning.A method for quantitation of the glucosyl ester of abscisic acid as the tetraacetate derivative by gas-liquid chromatography with an electron capture detector was developed. The level of beta-d-glycopyranosyl abscisate in Xanthium leaves increased from 3.6 nanomoles per gram fresh weight in turgid leaves to 22.9 nanomoles in leaves from plants subjected to seven wilting-recovery cycles. beta-d-glycopyranosyl abscisate in Xanthium leaves may be a stable end product of abscisic acid metabolism.

Journal Article↗