Concentration dependence of protein diffusion.
The concentration dependence of protein self-diffusion constants is described by a free volume diffusion theory which accounts for the effects of local protein concentration fluctuations.
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Internal cations, membrane current, and sodium inactivation gate closure in Myxicola giant axons.
Steady state to peak Na current ratio (INa,/INapeak) in Myxicola is greater, under some conditions, in internal Cs than in K, indicating less steady state inactivation in Csi. Csi effects are selective for steady state inactivation, with negligible effects on single-pulse inactivation time constants (Th). Mean Th ratios (Csi to Ki) were 1.04 and 1.02 at 0 and 10 mV. Two pulse inactivation time constants were also little affected. Inactivation is blocked in an all or none manner. Ki has little effect on steady state inactivation in the presence of inward INa, with INa/INapeak often declining to zero at positive potentials and independent of external Na concentration from 1/4 to 2/3 artificial sea water (ASW). Cs also has little effect at more negative potentials, but more with either more positive potentials or Na reduction, both reducing inward INa. K effects are evident when Na channel current is outward. A site in the current pathway when occupied selectively blocks inactivation gate closure. As occupancy does not depend significantly on potential, the site must not be very deep into the membrane field. Inactivation gates may associate with these sites on closure. The inactivated state may consist of a positively-charged structure occluding the inner channel mouth.
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Gating current kinetics in Myxicola giant axons. Order of the back transition rate constants.
Gating current, Ig, was recorded in Myxicola axons with series resistance compensation and higher time resolution than in previous studies. Ig at ON decays as two exponentials with time constants, tau ON-F and tau ON-S, very similar to squid values. No indication of an additional very fast relaxation was detected, but could be still unresolved. Ig at OFF also displays two exponentials, neither reflecting recovery from charge immobilization. Deactivation of the two I(ON) components may proceed with well-separated exponentials at -100 mV. INa tail currents at OFF also display two exponentials plus a third very slow relaxation of 5-9% of the total tail current. The very slow component is probably deactivation of a very small subpopulation of TTX sensitive channels. A -100 mV, means for INa tail component time constants (four axons) are 76 microseconds (range: 53-89 microseconds) and 344 microseconds (range: 312-387 microseconds), and for IOFF (six axons) 62 microseconds (range: 34-87 microseconds) and 291 microseconds (range: 204-456 microseconds) in reasonable agreement. INa ON activation time constant, tau A, is clearly slower than tau ON-F at all potentials. Except for the interval -30 to -15 mV, tau A is clearly faster than tau ON-S, and has a different dependency on potential. tau ON-S is several fold smaller than tau h. Computations with a closed2----closed1----open activation model indicated Na tail currents are consistent with a closed1----open rate constant greater than the closed2----closed1.
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Cytoplasmic hydrogen ion diffusion coefficient.
The apparent cytoplasmic proton diffusion coefficient was measured using pH electrodes and samples of cytoplasm extracted from the giant neuron of a marine invertebrate. By suddenly changing the pH at one surface of the sample and recording the relaxation of pH within the sample, an apparent diffusion coefficient of 1.4 +/- 0.5 x 10(-6) cm2/s (N = 7) was measured in the acidic or neutral range of pH (6.0-7.2). This value is approximately 5x lower than the diffusion coefficient of the mobile pH buffers (approximately 8 x 10(-6) cm2/s) and approximately 68x lower than the diffusion coefficient of the hydronium ion (93 x 10(-6) cm2/s). A mobile pH buffer (approximately 15% of the buffering power) and an immobile buffer (approximately 85% of the buffering power) could quantitatively account for the results at acidic or neutral pH. At alkaline pH (8.2-8.6), the apparent proton diffusion coefficient increased to 4.1 +/- 0.8 x 10(-6) cm2/s (N = 7). This larger diffusion coefficient at alkaline pH could be explained quantitatively by the enhanced buffering power of the mobile amino acids. Under the conditions of these experiments, it is unlikely that hydroxide movement influences the apparent hydrogen ion diffusion coefficient.
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Sweet is stable: glycosylation stabilizes collagen.
For most collagens, the melting temperature (T(m)) of the triple-helical structure of collagen correlates with the total content of proline (Pro) and 4-trans-hydroxyproline (Hyp) in the Xaa and Yaa positions of the -Gly-Xaa-Yaa- triplet repeat. The cuticle collagen of the deep-sea hydrothermal vent worm Riftia pachyptila, despite a very low content of Pro and Hyp, has a relatively high thermal stability. Rather than Hyp occupying the Yaa position, as is normally found in mammalian collagens, this position is occupied by threonine (Thr) which is O-glycosylated. We compare the triple-helix forming propensities in water of two model peptides, Ac-(Gly-Pro-Thr)(10)-NH(2) and Ac-(Gly-Pro-Thr(Galbeta))(10)-NH(2), and show that a collagen triple-helix structure is only achieved after glycosylation of Thr. Thus, we show for the first time that glycosylation is required for the formation of a stable tertiary structure and that this modification represents an alternative way of stabilizing the collagen triple-helix that is independent of the presence of Hyp.
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Genome methylation of the marine annelid worm Chaetopterus variopedatus: methylation of a CpG in an expressed H1 histone gene.
Hydrolysis by methylation-dependent restriction enzymes shows that the genomic DNA of the polychaete annelid worm Chaetopterus variopedatus is methylated. Electrophoretic analyses of the digestion products indicate that the degree of methylation is lower in adult tissues than in sperm and embryonic DNA. 5-Methylcytosine was identified by HPLC, absorption spectroscopy and mass spectrometry analyses of free bases obtained by acid hydrolysis of the DNA. An average value of 1.6% methylated cytosines was determined in sperm DNA. Partial methylation was also found in an actively expressed H1 histone gene. This is the first time that genomic DNA methylation is demonstrated to occur in a worm.
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Origin of octameric creatine kinases.
Mitochondrial creatine kinase (MiCK) occurs primarily as an octameric form localized in the mitochondrial intermembrane compartment in vertebrate tissues and echinoderm spermatozoa (both deuterostome groups). The octameric quaternary structure is thought to play important functional and enzyme targeting roles. We have found that the spermatozoa of the protostome polychaete Chaetopterus variopedatus contain three distinct isoenzymes of creatine kinase (CK) termed CK1, CK2 and CK3. CK3 appears to be present only in the sperm head/midpiece complex where mitochondria are restricted and has a subunit relative molecular mass (Mr) of 43.4 kDa. Gel permeation chromatography using Superdex 200HR showed that CK3 has a native Mr of 344.9 kDa indicating that this enzyme exists as an octamer. Electron micrographs of negatively stained CK3 preparations show structures which are virtually identical to those that have been seen for octameric vertebrate MiCK. The above observations show that CK3 from C. variopedatus displays great similarities to MiCKs from vertebrates and echinoderms. Octamerization of CK is not an advanced feature. The evolution of octameric subunit association is ancient and occurred prior to the divergence of protostomes and deuterostomes.
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The ciliate Orchitophrya cf. stellarum and other parasites and commensals of the northern Pacific seastar Asterias amurensis from Japan.
To identify the pathogens and possible biological control agents for the introduced seastar Asterias amurensis, we examined seastars from source populations in central and northern Japan. In particular, we sought the scuticociliate Orchitophrya cf. stellarum. The ciliate was found in male A. amurensis from five sites. We also found the caprellid amphipod Caprella astericola on A. amurensis and Distolasterias nipon from Nemuro Bay. The copepod Scottomyzon gibberum was found on A. amurensis from Usujiri and polychaete scaleworms Arctonoe vittata were found on A. amurensis from Murohama and Nemuro Bay. Of these parasites and commensals, Orchitophrya cf. stellarum is the most likely agent for biological control of A. amurensis in Australian waters; however, its ability to regulate seastar populations is uncertain.
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Polydora ciliata shell infestation in Tapes philippinarum Manila clam held out of the substrate in the Adriatic sea, Italy.
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Animals↗
Infestation of the clam Chione fluctifraga by the burrowing worm Polydora sp. nov. in laboratory conditions.
Burrowing worms that belong to Polydora spp. infest marine mollusks cultured worldwide, causing problems for production and marketing. The clam Chione fluctifraga is semi-cultured in Bahía Falsa, Baja California, NW Mexico, and some clams harbor burrowing worms. The present study was carried out to determine the identity of the worm species infesting the clam, the infesting process by cohabitation of infested and non-infested clams in aquaria with a variety of substrates (fine sand, gross sand, plastic bag used for clam culture, and aquarium without substrate) and turbulence conditions, and the occurrence of architomy phenomena in connection with infestation of the clam. The burrowing worm was considered as a nova species due to its singular limbate neurosetae and notosetae in the setiger 5, hooks in the setiger 6, eyes not present, and general pigmentation, among other characteristics. Infestation was similar in all substrates and turbulence conditions, but it was more abundant on clams previously infested than on those free of worms, showing a preferential settlement of worm infesting stages on pre-infested clams. Regeneration was observed in all segments of the worm: anterior (metastomium), medium, and posterior (prostomium); the complete regeneration time occurred in 40 days. This is the first record of architomy in a species of Polydora and this phenomenon could account for the increase of infestation intensity in pre-infested clams at the end of the study period. Infestation of clams by settling polichaete in the conditions studied, and the architomy process in this worm species, shows its great infesting capacity.
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Structural similarities and differences amongst neurofilaments.
Neurofilaments purified from cat, ox, Loligo and Myxicola nerve fibres are composed of different numbers of polypeptides with different molecular weights. Mammalian neurofilaments also differ from those of marine invertebrates by being about 20% larger in diameter. Despite the differences, X-ray diffraction patterns from all the neurofilaments indicate a common alpha-helical content with alpha-helices arranged in coiled-coils. The patterns from Myxicola neurofilaments also indicate a long-range periodicity along the length of these filaments which is of the order of 25.2 nm.
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Thick filaments and paramyosin of annelid muscles.
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Animals↗
Redistribution of intramembrane particles related to acetylcholine release by cholinergic synaptosomes.
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Acetylcholine↗
Polychaetes as indicators of environmental disturbance on subarctic tidal flats, Iqaluit, Baffin Island, Nunavut Territory.
The polychaetes of the tidal flats near the town of Iqaluit, Baffin Island were analysed along gradients of environmental disturbance resulting from human activity. Sources of environmental disturbance include a sewage lagoon, garbage sites; and an area of the tidal flat that is cleared by bulldozer. Sampling of the tidal flats included 300 biological sediment cores taken from 75 sites along seven transects. Environmental disturbance has resulted in four zones of polychaete communities with increasing distance. The heavily disturbed zone is closest to the disturbances and is devoid of polychaetes. The disturbed zone follows and is characterized by low diversity the result of increased densities of a few opportunistic species such as, Capitella 'capitata' sp. The moderately disturbed zone is characterized by increased species diversity due to organic enrichment from the disturbances. The undisturbed zone, located the furthest from the sources of disturbance, is characterized by moderate levels of diversity compared to the other three zones.
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Response of an estuarine benthic community to application of the pesticide carbaryl and cultivation of Pacific oysters (Crassostrea gigas) in Willapa Bay, Washington.
Oyster culture operations on the West coast of North America have developed into complete farming operations for the introduced Japanese oyster, Crassostrea gigas, which now covers vast areas of the intertidal landscape, particularly in Washington State where the pesticide carbaryl has also been used to control burrowing thalassinid shrimp for more than 30 years. Field experiments were conducted to examine the effects of these habitat modifications on the benthic community in Willapa, Bay Washington (124 degrees 06'W,46 degrees 24'N) where 50% of the state's oyster production occurs. Results indicated that the primary long-term effect of carbaryl application was removal of the two species of thalassinid shrimp (Neotrypaea californiensis and Upogebia pugettensis), which dominated the community at the start of the experiment and clearly influenced community composition themselves. Small peracarid crustaceans like the amphipods Corophium acherusicum and Eohaustorius estuarius experienced the most significant short-term mortalities, but generally recruited back to treated sites within 3 months, and were often more abundant on treated than untreated sites 1 year after carbaryl application. Results for molluscs were mixed, with no significant effect on Macoma spp, but a significant effect on the commensal clam Crytomya californica and mixed results for the cockle Clinocardium nutalli. Polychaetes were the least susceptible to carbaryl and with the exception of a short-term effect on oligochaetes, no significant negative effects were observed. The addition of oysters did not affect the infaunal community in this study, however greater abundance of epifaunal organisms like mussels, scaleworms, and the amphipod Amphithoe valida, which builds tubes in algae attached to shells, was observed. Carbaryl, which is currently applied to roughly 242 ha (< 6% of the intertidal) in Willapa Bay on an annual basis, has a variable but relatively short-term effect on the benthic community, which should be viewed in the context of other oyster culture operations like the addition of oysters themselves to a community often dominated by burrowing thalassinid shrimp which clearly control its dynamics.
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Incorporation of organic tritium (3H) by marine organisms and sediment in the severn estuary/Bristol channel (UK).
Discharges of tritium (3H) into the Severn estuary/Bristol Channel (UK) arise from the authorized release of wastes from nuclear power plants at Hinkley Point and Berkley/Oldbury and from the Nycomed-Amersham radiochemical plant, via the sewer system, at Cardiff. The wastes from the nuclear power plants probably consist almost entirely of 3H2O, whereas those from the radiochemical plant also include uncharacterized 3H labelled organic compounds. The total 3H concentrations in demersal fish and other benthic organisms in the vicinity of the Cardiff Eastern sewer outfall are significantly elevated compared to those observed around other UK nuclear establishments. Concentrations in filtered seawater were approximately 10 Bq kg(-1) whilst levels in surface sediment, seaweed (Fucus vesiculosis) and mussels (Mytilus edulis)/flounder (Platichthys flesus) were in the order of 6 x 10(2), 2 x 10(3), and 10(5) Bq kg(-1) (dry weight), respectively. Almost all the 3H found in sediment and biota were organically bound tritium (OBT). The high concentration in these materials, relative to that in seawater, is due to the presence of bioavailable organic 3H labelled compounds in the radiochemical waste. It is suggested that bioaccumulation of 3H by benthic organisms and demersal fish occurs primarily via a pathway of physico-chemical sorption/bacterial transformation of dissolved 3H labelled organic compounds into particulate organic matter, and subsequent transfer up a web of sediment dwelling microbes and meiofauna. Variations in 3H accumulation between individual organisms have been interpreted in terms of their different feeding behaviour. Relatively low concentrations were observed in the herbivorous winkle (Littorina littorea) and the pelagic Sprat (Spratus spratus) compared with other benthic organisms and demersal fish. The elevated 3H concentrations in seafood, due to bioaccumulation of OBT, have low radiological significance even for the local critical group of seafood consumers.
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Toxicological and chemical assessment of ordinance compounds in marine sediments and porewaters.
Toxicological and chemical studies were performed with a silty and a sandy marine sediment spiked with 2,6-dinitrotoluene (2,6-DNT), 2,4,6-trinitrophenylmethylnitramine (tetryl), or 2,4,6-trinitrophenol (picric acid). Whole sediment toxicity was analyzed by the 10-day survival test with the amphipod Ampelisca abdita, and porewater toxicity tests assessed macro-algae (Ulva fasciata) zoospore germination and germling growth, sea urchin (Arbacia punctulata) embryological development, and polychaete (Dinophilus gyrociliatus) survival and reproduction. Whole sediments spiked with 2,6-DNT were not toxic to amphipods. The fine-grained sediment spiked with tetryl was also not acutely toxic. The tetryl and picric acid LC50 values in the sandy sediment were 3.24 and 144 mg/kg dry weight, respectively. The fine-grained sediment spiked with picric acid generated a U-shaped concentration-response curve in the amphipod test, with increased survival both in the lowest and highest concentration. Grain-size distribution and organic carbon content strongly influenced the behavior of ordnance compounds in spiked sediments. Very low concentrations were measured in some of the treatments and irreversible binding and biodegradation are suggested as the processes responsible for the low measurements. Porewater toxicity varied with its sedimentary origin and with ordnance compound. The sea urchin embryological development test tended to be the least sensitive. Tetryl was the most toxic chemical in all porewater tests, and picric acid the least toxic. Samples spiked with 2,6-DNT contained a degradation product identified as 2-methyl-3-nitroaniline (also known as 2-amino-6-nitrotoluene), and unidentified peaks, possibly degradation products, were also seen in some of the picric acid- and tetryl-spiked samples. Degradation products may have played a role in observed toxicity.
Amphipoda↗
Toxicity of 1,4-dichlorobenzene in sediments to juvenile polychaete worms.
Investigation of sediment contamination associated with a marine sewage outfall in Victoria (BC, Canada) found elevated concentrations of 1,4-dichlorobenzene (1,4-DCB). Juvenile polychaete worm (Neanthes) growth was significantly reduced at or near the outfall, roughly corresponding to elevated 1,4-DCB concentrations. There are few data on 1,4-DCB toxicity to marine organisms and no published literature on its toxicity to benthic marine organisms. To determine whether reduced polychaete growth (measured as dry weight) was due to 1,4-DCB exposure, a laboratory investigation was conducted. Uncontaminated marine sediment was spiked with 1,4-DCB and juvenile Neanthes were exposed in 20-d sublethal toxicity tests. There were no adverse effects on survival at any test concentration; mean survival was 80-100%. Statistically significant decreases in average dry weight only occurred at the highest 1,4-DCB concentration (19,900 microg/kg, dry weight); this represented a 1,4-DCB concentration more than 10 times higher than previously measured at the outfall (1,710 microg/kg, dry weight). There were no adverse effects on survival or dry weight at the range of concentrations previously measured in sediments from the vicinity of the outfall.
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