Search

Methods for estimating the absolute polyphenol content of the brown algae Ascophyllum nodosum (L.) Le Jol. and Fucus vesiculosus (L.) have been developed and tested. Polyphenols were extracted almost quantitatively from Ascophyllum nodosum using aqueous acetone, whereas this procedure was somewhat less efficient with Fucus vesiculosus. Colorimetric methods based on the Folin-Denis reagent, Brentamine Fast Red 2G Salt, and vanillin-H2SO4 were applied to acetone-free extracts for determination of polyphenol content relative to suitable reference compounds. Gravimetric methods based on hide powder and on haemoglobin were employed to derive ‘estimation factors’ (EFs) which allow calculation of the absolute polyphenol content from the relative polyphenol content. The values calculated for absolute polyphenol content are considered to be reasonably accurate, despite imprecisions in the methods and despite often large standard deviations, and re-emphasize the potential physiological and ecological significance of brown algal polyphenols. Although the precise EFs calculated here are not valid for other brown algae, the methods are considered to be generally applicable to other Phaeophyceae.

The causative agent of toxicity in cultured mussels from a localized area of eastern Prince Edward Island has been identified as domoic acid, a neuroexcitatory amino acid. The toxin was isolated by a number of different bioassay-directed separation techniques including high-performance liquid chromatography, high-voltage paper electrophoresis, and ion-exchange chromatography, and characterized by a number of spectroscopic techniques including ultraviolet, infrared, mass spectrometry, and nuclear magnetic resonance. The isolation and purification methods are described in detail and some new analytical data for domoic acid are reported. Keywords: shellfish toxin, domoic acid, neurotoxin, bioassay-directed analysis.

When freshly collected thalli of Ascophyllum nodosum (L.) Le Jol. were allowed to stand in batches of filtered sea water for 1 to 4 h at 10 ± 2° C , polyphenolic substances were released into the contact media. Polyphenol release during photosynthesis far exceeded dark-release, and rates of release were greater during the initial period of the experiment (whether light or dark) than during subsequent light or dark periods, respectively. The release could be ‘turned on’ and ‘turned off’ by successive irradiation and darkening of a single sample, demonstrating that much of the release is exudation in the strict sense, not merely a symptom of stress, shock or injury. No light intensity threshold for polyphenol exudation could be detected, and an apparent ‘saturation’ was seen at intensities above 9 × 10 2 W m −2 . Rates of light-mediated release of polyphenols under these protected laboratory conditions were 9.2 ± 3.6 μg · g −1 dry matter·h −1 .

High-molecular-weight, nondialyzable polyphloroglucinols from the marine brown algae Ascophyllum nodosum and Fucus vesiculosus (Phaeophyceae, Fucales) chelate the divalent metal ions Sr2+, Mg2+, Ca2+, Be2+, Mn2+, Cd2+, Co2+, Zn2+, Ni2+, Pb2+ and Cu2+ in weakly acidic aqueous solution. Values of the relative selectivity coefficients k have been calculated relative to Mn2+; the ionotropic series with the algal polyphenols are similar to those with catechol and pyrogallol. These studies may serve as a guide for the examination of metal chelation by brown algal polyphenols both in situ and after exudation into seawater.

The content of extractable polyphenols in the brown algae Ascophyllum nodosum (L.) Le Jol. and Fucus vesiculosus (L.) was measured at ≈28-day intervals for one year. Colorimetric methods based on the Folin-Denis, Brentamine, and vanillin-H2SO4 reactions were used to estimate relative contents of polyphenols, and these values were converted to absolute contents using the gravimetric method introduced earlier. Relatively little error was introduced by variations in the qualitative composition of the extracted polyphloroglucinols. There appeared to be a significant temporal correlation between polyphenol content and the reproductive state of the algae. The content of polyphenols in A. nodosum was at a minimum (≈9–10% of dry matter) during the period of maximum fruit body shedding (late May), and reached a maximum (≈12–14% of dry matter) during the ‘winter season’. In F. vesiculosus, the minimum (≈8–10% of dry matter) was one to two months later, just before the period of maximum fertility, and thereafter rose to a maximum (≈11–13% of dry matter) during the period of sterility. These results furthermore suggest that the bulk of the polyphenols are not readily accessible as reserve components, and indicate that modifications may be needed in the ‘chemical defense’ and ‘waste product’ hypotheses concerning the significance of brown algal polyphenols.

Poor correlation exists between levels of total phenols and of ultraviolet-absorbing materials in extracts of the brown algae Ascophyllum nodosum (L.) Le Jol. and Fucus vesiculosus L. The uv extinction sp coefficient of a major exuded polyphenol fraction increases rapidly and markedly in sea-water solution. Consequently, uv spectrophotometry cannot provide a useful measure of either total polyphenols or total organic matter in extracts or exudates from these brown algae. Moreover, considerable errors may be introduced when uv spectrophotometry is used to estimate dissolved organic matter in sea water which receives significant exudation from brown macrophytes.

1H and 13C nuclear magnetic resonance spectroscopy of the high-molecular-weight (> 104) polyphenol fraction from Fucusvesiculosus, before and after acetylation or methylation, demonstrates that it is composed of phloroglucinol units (48.9 ± 3.5%) and 2,2′,4,4′,6,6′-hexahydroxybiphenyl, 2,2′,2″,4,4′,4″,6,6′,6″-nonahydroxy-1,1′:3′1″-terphenyl, and related quater-and polyphenyl groups (together, 51.1 ± 3.5%) linked by ether bonds, each bond involving the carbinol and methine carbons of different units and the formal loss of a molecule of hydrogen. The polymer is highly branched, with ca. 20 – 22% of the constituent units being chain termini; there is no evidence for large rings of units. Approximately 5 – 10% of the phloroglucinol and 54 – 82% of the directly bonded units occur on the exterior of the molecule, whereas the interior backbone consists predominantly of ether-linked phloroglucinol units.

The green alga Spirogyra sp. accumulates tetra- through undecagalloylglucosyl gallotannins. The hexa- through undecagalloylglucoses are predominantly based on 1,2,3,4,6-penta-O-galloylglucose, whereas the major pentagalloylglucose is 3-O-digalloyl-1,2,6-tri-O-galloyl-β- D -glucose.

Vanadium tetrachloride-catalyzed dimerization of 3,5-dimethoxyphenol yields the key intermediate 2,2′,6,6′-tetra-methoxy-4,4′-dihydroxybiphenyl, from which the octamethyl ether of the brown algal metabolite "fucophlorethol A" (2,2′,4,6,6′-pentamethoxy-4′-(2,4,6-trimethoxyphenoxy)biphenyl, 6) is synthesized by Ullmann condensation and Hakomori methylation.

Fucus vesiculosus accumulates polyphenols consisting of four-seven phloroglucinol units. These ‘oligomers’ are primarily ether-linked; no evidence could be found for entirely phenyl-linked ‘oligomers’ having more than four phloroglucinol units, nor for any ‘oligomer’ with more than three phenyl linkages, nor for vicinally trihydroxylated polyphloroglucinols.

Phenol sulfate esters have been analyzed by paired-ion reversed-phase high-performance liquid chromatography. The method provided direct, rapid chromatography of phenol sulfates in crude extracts of the red alga Polysiphonai lanosa (2,3-dibromo-4,5-dihydroxybenzyl alcohol 1′,4-disulfate), of the brown alga Ascophyllum nodosum (1,2,3,5-tetrahydroxybenzene 2,5-disulfate), and in rat urine (resorcinol mono- and disulfates). Detector response (254 nm) was linear within the approximate range from 30–125 ng to 5–10 μg. Semipreparative scale chromatography provided sufficient amounts of purified phenol sulfates for further analysis by paper electrophoresis.

Does the “universal tree” based on small-subunit ribosomal RNA sequences show the phylogenetic relationship of all modern organisms? The answer is “yes” only if all these rRNAs are orthologous. Herein I argue that the major rRNA lineages (e.g. eubacterial, one or more archaebacterial and eukaryotic nucleocytoplasmic) probably arose from a divergent population of rRNAs in the progenote, antedating the universal common ancestral organism. Thus the major lineages of rRNA are probably not orthologous, but paralogous. The extrapolated date for the origin of the common ancestral small-subunit rRNA (3.6–4.7 × 10 9 years ago) is consistent with major rRNA lineages being paralogous. This perspective on the early evolution of genes and organisms rationalizes the presence of unexpected ribosomal characters in microsporidia, and bears on xenogenous and endogenous theories of the origin of organelles in eukaryotis.

Alcoholic extracts of Fucus vesiculosus contain small quantities of low molecular weight polyphenols derived from phloroglucinol and 2,2′,4,4′,6,6′-hexahydroxybiphenyl. 1H and 13C nmr were used to identify two of these as 4-(2′′,4′′,6′′-trihydroxyphenoxy)-2,2′,4′,6,6′-pentahydroxybiphenyl and 4-(2′′-(2′′′,4′′′,6′′′-trihydroxyphenoxy)-4′′,6′′-dihydroxyphenoxy)-2,2′,4′, 6,6′-pentanydroxybiphenyl.

A series of vanillin-reactive compounds has been isolated from extracts of the brown alga Fucus vesiculosus. The first three members of this series have been examined by mass spectrometry, nuclear magnetic resonance spectrometry, and chemical methods, and were shown to be phloroglucinol, its phenyl-linked dimer, and a trimer also composed of phloroglucinol units. Evidence is presented for the presence of tetrameric, pentameric, and hexameric phloroglucinol derivatives. Polymeric vanillin-reactive compounds were isolated and separated into two fractions. Oxidative hydrolysis of the major one produced phloroglucinol and related oligomers in low yields. The classical vanillin reactivity of physodes may thus be attributed to phloroglucinol-containing derivatives, as originally proposed by Crato (Crato, E.: Ber. Dtsch. Bot. Ges. 10, 295–302 (1892)).

Many efforts at standardising terminologies within the biological domain have resulted in the construction of hierarchical controlled vocabularies that capture domain knowledge. Vocabularies, such as the PSI-MI vocabulary, capture both deep and extensive domain knowledge, in the OBO (Open Biomedical Ontologies) format. However hierarchical vocabularies, such as PSI-MI which are represented in OBO, only represent simple parent-child relationships between terms. By contrast, ontologies constructed using the Web Ontology Language (OWL), such as BioPax, define many richer types of relationships between terms. OWL provides a semantically rich structured language for expressing classes and sub-classes of entities and properties, relationships between them and domain-specific rules or axioms that can be applied to extract new information through semantic inferencing. In order to fully exploit the domain knowledge inherent in domain-specific controlled vocabularies, they need to be represented as OWL-DL ontologies, rather than in formats such as OBO. In this paper, we describe a method for converting OBO vocabularies into OWL and class instances represented as OWL-RDF triples. This approach preserves the hierarchical arrangement of the domain knowledge whilst also making the underlying parent-child relationships available to inferencing engines. This approach also has clear advantages over existing methods which incorporate terms from external controlled vocabularies as literals stripped of the context associated with their place in the hierarchy. By preserving this context, we enable machine inferencing over the ordered domain knowledge captured in OBO controlled vocabularies

The nucleotide sequence of a 23S rRNA gene of Campylobacter coli VC167 was determined. The primary sequence of the C. coli 23S rRNA was deduced, and a secondary-structure model was constructed. Comparison with Escherichia coli 23S rRNA showed a major difference in the C. coli rRNA at approximately position 1170 (E. coli numbering) in the form of an extra sequence block approximately 147 bp long. PCR analysis of 31 other strains of C. coli and C. jejuni showed that 69% carried a transcribed spacer of either ca. 147 or ca. 37 bp. Comparison of all sequenced Campylobacter transcribed spacers showed that the Campylobacter inserts were related in sequence and percent G+C content. All Campylobacter strains carrying transcribed spacers in their 23S rRNA genes produced fragmented 23S rRNAs. Other strains which produced unfragmented 23S rRNAs did not appear to carry transcribed spacers at this position in their 23S rRNA genes. At the 1850 region (E. coli numbering), Campylobacter 23S rRNA displayed a base pairing signature most like that of the beta and gamma subdivisions of the class Proteobacteria, but in the 270 region, Campylobacter 23S rRNA displayed a helix signature which distinguished it from the alpha, beta, and gamma subdivisions. Phylogenetic analysis comparing C. coli VC167 23S rRNA and a C. jejuni TGH9011 (ATCC 43431) 23S rRNA with 53 other completely sequenced (eu)bacterial 23S rRNAs showed that the two campylobacters form a sister group to the alpha, beta, and gamma proteobacterial 23S rRNAs, a positioning consistent with the idea that the genus Campylobacter belongs to the epsilon subdivision of the class Proteobacteria.