An electron microscope and autoradiographic study of the calciferous glands of the earthworm, Lumbricus terrestris.
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Immunoreactive platelet-derived growth factor-AB and transforming growth factor-beta 1 were demonstrated in invertebrate and vertebrate immunocytes by an immunocytochemical procedure. These factors are only present in phagocytic cells among invertebrate immunocytes, whereas in vertebrate immunocytes they are found in monocytes, granulocytes, lymphocytes, thrombocytes and platelets. These results, in agreement with previous reports, represent further evidence in favour of the hypothesis that Nature has followed a conservative strategy in using a common pool of signal molecules that have been highly conserved throughout evolution.
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Electrochemical determination of metallothionein (MT) is widely used for environmental studies. This article describes the development and optimization of the procedure for the quantification of metallothionein by square wave cathodic stripping voltammetry. The determination is based on the complexation of cisplatin and MT and the subsequent reduction of the complexes at the electrode. In order to achieve the highest possible sensitivity and resolution of the peak, an optimization of the experimental parameters has been carried out using experimental design methodology (response surface). Seven chemical and physical parameters, namely, pH, cisplatin concentration, buffer concentration, deposition potential, square wave frequency, amplitude of pulse, and step potential, have been optimized to give 9.0, 5.9 microM, 0.65 M, -0.2 mV, 229 Hz, 46 mV, and 2 mV, respectively. Method characterization has been performed, leading to a detection limit of 0.1 microg L(-1). Quantification of MT in polychaetes and comparison with the modified Brdicka procedure were also carried out.
The ice worm, Mesenchytraeus solifugus, is the largest glacially obligate metazoan and among only a few metazoan species that complete their life cycle at 0 degrees C. We conducted a large-scale sequencing analysis of cDNAs isolated from ice worm anterior segments. Sequence comparisons among an available group of ice worm, arthropod, chordate, and nematode homologues suggest that ice worms encode proteins that are less bulky, are less polar, and contain fewer charged residues. Also, subunits of the catalytic F1 ATP synthase complex appear to have diverged more rapidly than other ice worm genes examined, suggesting a role in cold-temperature adaptation. Modeling of F1 ATP synthase beta and gamma subunits identified nonconservative, ice worm-specific amino acid substitutions at subunit contact sites and at sites proximal to the catalytic site.
Giant extracellular hexagonal bilayer hemoglobin (HBL-Hb), found only in annelids, is an approximately 3500-kDa heteropolymeric structure involved in oxygen transport. The HBL-Hbs are comprised of globin and linker chains, the latter being required for the assembly of the quaternary structure. The linker chains, varying in size from 225 to 283 amino acids, have a conserved cysteine-rich domain within their N-terminal moiety that is homologous to the cysteine-rich modules constituting the ligand binding domain of the low-density lipoprotein receptor (LDLR) protein family found in many metazoans. We have investigated the gene structure of linkers from Arenicola marina, Alvinella pompejana, Nereis diversicolor, Lumbricus terrestris, and Riftia pachyptila. We found, contrary to the results obtained earlier with linker genes from N. diversicolor and L. terrestris, that in all of the foregoing cases, the linker LDL-A module is flanked by two phase 1 introns, as in the human LDLR gene, with two more introns in the 3' side whose positions varied with the species. In addition, we obtained 13 linker cDNAs that have been determined experimentally or found in the EST database LumbriBASE. A molecular phylogenetic analysis of the linker primary sequences demonstrated that they cluster into two distinct families of linker proteins. We propose that the common gene ancestor to annelid linker genes exhibited a four-intron and five-exon structure and gave rise to the two families subsequent to a duplication event.
Tertiary structures of proteins are conserved better than their primary structures during evolution. Quaternary structures or subunit organizations, however, are not always conserved. A typical case is found in hemoglobin family. Although human, Scapharca, and Urechis have tetrameric hemoglobins, their subunit contacts are completely different from each other. We report here that only one or two amino acid replacements are enough to create a new contact between subunits. Such a small number of chance replacements is expected during the evolution of hemoglobins. This result explains why different modes of subunit interaction evolved in animal hemoglobins. In contrast, certain interactions between subunits are necessary for cooperative oxygen binding. Cooperative oxygen binding is observed often in dimeric and tetrameric hemoglobins. Conformational change of a subunit induced by the first oxygen binding to the heme group is transmitted through the subunit contacts and increases the affinity of the second oxygen. The tetrameric hemoglobins from humans and Scapharca have cooperativity in spite of their different modes of subunit contact, but the one from Urechis does not. The relationship between cooperativity and the mode of subunit contacts is not clear. We compared the atomic interactions at the subunit contact surface of cooperative and non-cooperative tetrameric hemoglobins. We show that heme-contact modules M3-M6 play a key role in the subunit contacts responsible for cooperativity. A module was defined as a contiguous peptide segment having compact conformation and its average length is about 15 amino acid residues. We show that the cooperative hemoglobins have interactins involving at least two pairs of modules among the four heme-contact modules at subunit contact.
Implementation of advanced wastewater treatment at the two municipal wastewater-treatment plants for Indianapolis, Indiana, resulted in substantial improvement in the quality of the receiving stream and significant changes in the benthic-invertebrate community. Diversity, similarity, and biotic indices were compared to determine which indices best reflected changes in the composition of the biota in the river. None of the indices perfectly reflected the changes in river quality or community structure. Similarity indices, especially percentage similarity, exhibit the most promise of the three classes of indices. Diversity indices were least useful, wrongly indicating that water quality deteriorated after the upgrade of the wastewater-treatment plants. The most descriptive tool in analyzing the data was the percentage of Ephemeroptera, Plecoptera, and Trichoptera (EPT) taxa present. Using a mixture of indices and other analytical tools, such as EPT, in the analysis of biological data will ensure the most effective investigations of water quality.
The chronic effects of the herbicide diuron on survival and reproduction of Daphnia pulex, and survival and growth of the amphipod Hyalella azteca, the midge Chironomus tentans, juvenile and embryo/larval fathead minnows, Pimephales promelas, annelid worms, Lumbriculus variegatus, and snails, Physa gyrina, were determined in laboratory static and static-renewal tests. D. pulex 96-h and 7-day LC50 values were 17.9 and 7.1 mg/L; 7-day LOAEL and NOAEL values based on mortality and reproduction were 7.7 and 4.0 mg/L. H. azteca 96-h and 10-day LC50 values were 19.4 and 18.4 mg/L; 10-day LOAEL and NOAEL values based on survival and reduced weight were 15.7 and 7.9 mg/L. C. tentans 10-day LC50 value was 3.3 mg/L; 10-day LOAEL and NOAEL values based on growth were 7.1 and 3.4 mg/L, and 3.4 and 1.9 mg/L based on mortality. Juvenile fathead minnows had a 10-day LC50 of 27.1 mg/L and 10-day LOAEL and NOAEL values based on growth of 3.4 and <3.4 mg/L. The fathead minnow embryo-larval test had a 7-day LC50 value of 11.7 mg/L and 7-day LOAEL and NOAEL values based on reduced growth of 8.3 and 4.2 mg/L. L. variegatus had 10-day LOAEL and NOAEL values based on reduced weight of 3.5 and 1.8 mg/L. P. gyrina had 10-day LOAEL and NOAEL values based on reduced weight of 22.8 and 13.4 mg/L. Laboratory effects concentrations were higher that those found in normal field application situations, except in areas of localized pooling after recent herbicide applications, indicating that there would probably be little harm to these fish and invertebrates from diuron exposure in the field.
An echiuroid species, Urechis unicinctus, was surveyed for Hox genes using polymerase chain reaction with homeobox-specific degenerate primers. We identified nine distinct homeodomain-containing gene fragments. These nine fragments were classified by comparative analysis. This analysis revealed that this echiuroid possessed at least three Hox genes from the anterior group, five from the central group, and one from the posterior group.
Orthologs of the Drosophila gap gene hunchback have been isolated so far only in protostomes. Phylogenetic analysis of recently available genomic data allowed us to confirm that hunchback genes are widely found in protostomes (both lophotrochozoans and ecdysozoans). In contrast, no unequivocal hunchback gene can be found in the genomes of deuterostomes and non-bilaterians. We cloned hunchback in the marine polychaete annelid Platynereis dumerilii and analysed its expression during development. In this species, hunchback displays an expression pattern indicative of a role in mesoderm formation and neurogenesis, and similar to the expression found for hunchback genes in arthropods. These data suggest altogether that these functions are ancestral to protostomes.
Gutless tube-dwelling worms of pogonophorans (also known as frenulates) and vestimentiferans depend on primary production of endosymbiotic bacteria. The endosymbionts include thiotrophs that oxidize sulfur for autotrophic production and methanotrophs that oxidize and assimilate methane. Although most of the pogonophoran and vestimentiferan tube worms possess single thiotrophic 16S rRNA genes (16S rDNA) related to gamma-proteobacteria, some pogonohorans are known to bear single methanotroph species or even dual symbionts of thiotrophs and methanotrophs. The vestimentiferan Lamellibrachia sp. L1 shows symbiotic 16S rDNA sequences of alpha-, beta-, gamma-, and epsilon-proteobacteria, varying among specimens, with RuBisCO form II gene (cbbM) sequences related to beta-proteobacteria. An unidentified pogonophoran from the world's deepest cold seep, 7326-m deep in the Japan Trench, hosts a symbiotic thiotroph based on 16S rDNA with the RuBisCO form I gene (cbbL). In contrast, a shallow-water pogonophoran (Oligobrachia mashikoi) in coastal Japan Sea has a methanotrophic 16S rDNA and thiotrophic cbbL, which may suggest the feature of type X methanotrophs. These observations demonstrate that pogonophoran and vestimentiferan worms have higher plasticity in bacterial symbioses than previously suspected.
In this paper the author presents a mathematical model of a device that can be seen as a segment of an artificial worm (following the paradigm "earthworm") and as an artificial muscle as well. Confining considerations to statics, the model shows up as an ordinary parameter-dependent boundary value problem. It is tackled numerically in various particular forms by means of Maple and thus gives a good view of the segment's behavior during inflation and under longitudinal load. Segments of maximal volume present a useful preliminary stage of the investigations.
This paper summarizes sampling results from NOAA's National Status and Trends (NS&T) Program for marine environmental quality in Delaware Bay. A stratified-random design was used to determine the spatial extent of sediment contamination and toxicity in Delaware Bay from offshore stations in the coastal zone, the lower estuary, the upper estuary, the fresh/salt mixing zone, and tidal fresh areas. Sediment samples were taken for chemical analyses of major classes of environmental contaminants, a suite of toxicity bioassays, and benthic macrofaunal community assessment to identify patterns of resident species. The tidal-fresh areas and portions of the mixing zone of the study area were heavily contaminated. Contaminant concentrations were frequently above the 90th percentile of EMAP Virginian Province levels. PAHs in the sediment were higher than previously documented, with a major component of PAHs being pyrogenic in origin. Bioassay results were highly variable. Toxicity and contaminant levels were correlated when aggregated into indices, but were only marginally correlated with benthic community impacts. High and low abundance stations were found in all zones. Most of the tidal fresh stations were dominated by pollution tolerant species. Species diversity and abundance were generally lowest in the fresh/salt mixing zone.
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