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Structure and composition of ferritin cores isolated from human spleen, limpet (Patella vulgata) hemolymph and bacterial (Pseudomonas aeruginosa) cells.

Ferritin cores isolated from human spleen, limpet (Patella vulgata) hemolymph and bacterial (Pseudomonas aeruginosa) cells have been investigated by high resolution transmission electron microscopy, electron diffraction and chemical analysis. Hemosiderin particles isolated from thalassemic spleens also have been studied. The results show that there is a marked difference in structure and composition of the biomineral phases. Human ferritin and hemosiderin particles are single domain crystals of hydrated iron (III) oxide (ferrihydrite). Lattice fringes were low in contrast and often discontinuous within the central regions of the core. Heat treatment of human ferritins results in a 5 A shrinkage in particle size and an increase in the single crystalline nature of the core. In contrast, lattice images and electron diffraction of limpet and bacterial cores show no evidence of long-range crystallographic order. Chemical analysis indicates a high inorganic phosphate (Pi) (Fe/Pi = 1.71) content in bacterial ferritin compared with human ferritin (thalassemic) (Fe/Pi = 21.0). The high Pi content of bacterial ferritin suggests a hydrated amorphous iron (III) phosphate mineral core. Structural disorder within the limpet and bacterial cores may be associated with increased Pi content and increased oxidation in Fe(II), resulting in rapid mineral deposition. Growth of the iron (III) oxide cores in human ferritin is discussed on the basis of high resolution electron microscopy results.

Animals↗

The stringlike genes of the limpet Patella vulgata.

As a first step in analyzing the function of a cdc25 homolog during the embryonic development of Patella vulgata (Pv), genomic clones encoding these stringlike proteins (Stl) were isolated and characterized. These clones belong to four groups which are derived from different regions of the Pv genome. As the sequences of Stl genes from two of these groups are almost identical, we suggest that these genes represent copies of the same gene. The Stl3 gene, which has been analyzed in detail, consists of four exons separated by three introns. Its sequence encodes a 250-amino-acid protein with a calculated weight of 28 kDa. The Stl protein contains regions conserved in all other cdc25 proteins. Stl messengers are not stored maternally in Pv oocytes and Stl transcription only starts after the first embryonic cleavages.

Amino Acid Sequence↗

Organiser role of the stem cell of the mesoderm in prototroch patterning in Patella vulgata (Mollusca, Gastropoda).

During early development of the gastropod mollusc Patella vulgata, the stem cell of the mesoderm (3D-macromere) is induced. As a result of this induction, the embryo becomes dorsoventrally organised. At about the same time in development, ciliated cells, so-called trochoblasts, are formed. Later in development, some trochoblasts deciliate and, together with the ciliated trochoblast, form the dorsoventrally organised prototroch, the locomotory organ of the larva. In order to study the role of the 3D-macromere in the specification of trochoblasts and in the induction of the dorsoventral organisation of the prototroch, induction of 3D has been prevented in various ways. it is shown that preventing 3D-induction results in the formation of a radially symmetrical prototroch. The trochoblasts of all four quadrants developed like corresponding trochoblasts of the A-quadrant. Somewhere between 30 and 120 min after fifth cleavage the 3D-macromere induces the formation of specific trochoblasts and organises the dorsoventral pattern of the prototroch. Besides a role of the 3D-macromere, a role of other cells has been demonstrated in the conditionally specified deciliation of trochoblasts.

Animals↗

Putative midkine family protein up-regulation in Patella caerulea (Mollusca, Gastropoda) exposed to sublethal concentrations of cadmium.

A cDNA sequence of a putative midkine (MK) family protein was identified and characterised in the mollusc Patella caerulea. The midkine family consists of two members, midkine and pleiotrophin (PTN), and it is one of the recently discovered cytokines. Our results show that this putative midkine protein is up-regulated in specimens of P. caerulea exposed to sublethal cadmium concentrations (i.e. 0.5 and 1 mg l(-1) Cd) over a 10-day exposure period. Semiquantitative RT-PCR and quantitative Real time RT-PCR estimations indicate elevated expression of midkine mRNA in exposed specimens compared to controls. Moreover, RT-PCR Real time values were higher in the viscera (here defined as the part of the soft tissue including digestive gland plus gills) than in the foot (i.e. foot plus head plus heart) of the limpets. At present, information on the functional signalling significance of the midkine family proteins suggests that the up-regulation of P. caerulea putative midkine family protein is a distress signal likely with informative value on health status of the organism and with potential prognostic capability.

Amino Acid Sequence↗

Brachidontes variabilis and Patella sp. as quantitative biological indicators for cadmium, lead and mercury in the Lebanese coastal waters.

The mussel, Brachidontes variabilis, and the limpet, Patella sp., were used as indicators to monitor cadmium, lead and mercury concentrations along the Lebanese coast. Studies were carried out in order to define the best strategy for assessing and minimizing the effects of size and physiological condition on the metal contents of the molluscs, and corrective models were constructed. Metal concentrations in surface water were measured to estimate bioconcentration factors (BCFs). The BCFs varied from 8.3 x 10(3) to 3.4 x 10(4), from 7.5 x 10(3) to 8.0 x 10(3) and from 3.0 x 10(4) to 3.2 x10(4), for Cd, Pb and Hg, respectively. For limpets, BCFs varied from 1.7 x 10(4) to 7.4 x 10(4) for Cd, from 2.5 x 10(3) to 6 x 10(3) for Pb and remained fairly constant at around 10(4) for Hg. The highest BCFs were associated with lowest contamination levels. The results of the geographical survey exhibited a similar large-scale spatial pattern for the two species and followed the metal concentration distributions measured in the waters.

Animals↗

Time-of-day effect on patella tendon stiffness alters vastus lateralis fascicle length but not the quadriceps force-angle relationship.

AIM: To examine the time-of-day (TOD) effect on torque-force/angle, fibre length (FL), tendon stiffness (K), stress, and strain using the quadriceps muscle-tendon complex as a model. METHODS: Twelve healthy young men (aged 27+/-2.0 years) were studied at AM (7h45) and PM (5h45). Maximal isometric contractions were carried out on an isokinetic dynamometer, with real-time recordings of vastus lateralis (VL) FL and patella tendon K using B-mode ultrasonography. Percutaneous electrical twitch doublets superimposed on maximal torque were used to test for muscle activation capacity (AC). RESULTS: At PM, torque and force increased by 16+/-3.0% (P<0.01) over 30-90 degrees knee angles. Where the load was standardised (at 250N) in order to discriminate between torque generation capacity and tendon K changes, PM relative to AM, there were 8% and 13% (P<0.01) reductions in relaxed and contracted FL, respectively. Average K decreased by 21% (P<0.001) and the maximal stress and strain were increased at PM by 11% and 16%, respectively (P<0.01). No TOD effect on AC was seen. CONCLUSION: The quadriceps torque or force-angle relationships shift upwards at PM vs. AM, with no shift in the position of the optimal knee angle. This torque or force increase appears not to be centrally modulated. Although K decreases with TOD thereby potentially shortening the working length of the sarcomeres, these changes overall do not affect the ability of the muscle to produce greater torque in the evening.

Adult↗

Food supply, grazing activity and growth rate in the limpet Patella vulgata L.: a comparison between exposed and sheltered shores.

The limpet Patella vulgata L. is an important microphagous grazer on intertidal rocky shores of north-west Europe, occurring across the wave exposure gradient. Groups of P. vulgata were selected at mid-tide level of two exposed shores and two sheltered, fucoid dominated shores on the Isle of Man, British Isles, and manipulated to equivalent densities and population structure. The level of grazing activity and growth rate were determined over a 1-year period. At the same time, the abundance of epilithic microalgae, measured as the concentration of chlorophyll a, was determined as an estimate of food supply. Microagal abundance showed a seasonal pattern in both exposed and sheltered conditions, with higher levels in winter compared to summer. In both seasons, the microalgal resource was more abundant on the sheltered shore studied. The level of grazing activity in P. vulgata showed a seasonal pattern on the exposed but not the sheltered shores. Averaged over the year, grazing activity on the exposed shores was over double that on sheltered shores. Thus, in sheltered conditions, food supply for limpets was high and grazing activity low; in exposed conditions, food supply was low and grazing activity high. The growth rate of P. vulgata, measured as increase in shell length, showed no significant difference between exposed and sheltered shores. Growth rate was also determined in P. vulgata at natural densities. Although the overall density declined with decreasing exposure to wave action, the density per unit area of grazeable substance was higher in shelter. In these populations, the mean growth rate was over twice as high on exposed compared to sheltered shores.

Journal Article↗

Levels of polychlorinated biphenyls in Patella piperata from the coast of Fuerteventura (Canary Islands, Spain).

Concentration of 51 PCB congeners were determined in the visceral mass of the endemic limpet Patella piperata, sampled in four stations in the coast of Fuerteventura (Canary Islands, Spain) during 1991-1995, and evaluated in order to assess the origin of their contamination and the possibility of this limpet being used as a bioindicator organism for PCB contamination. PCB extracted from samples were fractionated on Florisil columns and analysed by capillary gas chromatography with flame ionisation detector. Principal Component, Factor and Cluster Analysis were used to study the patterns and relationships among PCBs.

Animals↗

Minor and trace elements in the shell of Patella aspera (Röding 1798).

Specimens of the limpet Patella aspera were collected from a clean, coastal marine site and a contaminated estuary on the south coast of Portugal. The shells were analysed individually for their minor (Mg, Sr) and trace element (Fe, Mn, Zn) content. Mean concentrations of these elements in the shell of P. aspera were 4651, 1318, 35.8, 29.9 and 5.5 microg g(-1), respectively. The elemental concentrations exhibited both a marked intra- and inter-population variability. Despite the variability within individual populations, significant differences in the trace element composition were apparent between the shells taken from the two sites. Small shells (< 2 g) provided the best resolution between sites for both manganese and iron. Differences in zinc were best resolved for larger shells. The shell of P. aspera has an extraordinarily high magnesium concentration, which is insensitive to gross salinity differences, and a trace metal assemblage that can be interpreted in terms of environmental exposure. On these grounds, it is recommended that the shell of P. aspera is a tissue for potential use in environmental trace metal monitoring.

Animals↗

Partitioning of trace metals between soft tissues and shells of Patella aspera.

This paper describes the partitioning of iron, manganese, zinc, copper, cadmium, cobalt, nickel and lead between the soft tissues and the shells of the gastropod mollusc Patella aspera. Specimens of the limpet P. aspera were collected from a clean coastal marine site (CMS) and from an estuarine contaminated site (ECS) on the south coast of Portugal. Fe and Zn concentrations were significantly lower on both populations and less variable in the shells than in the soft tissues. In contrast Mn concentrations were higher in the shells than in the soft tissues at ECS. The partitioning factor (PF), defined as the ratio between the mean metal concentrations in soft tissues and in the shells, was maximum for iron and minimum for manganese. Between the soft tissues and the shells, only manganese at ECS recorded a significant relationship between metal concentrations, suggesting that the mechanism that controls the accumulation of this essential metal in both tissues was different from the others. From the obtained data, in biomonitoring sewage contamination studies, shells of P. aspera can be considered as a good indicator for Mn while the soft tissues for Zn. As copper, cadmium, nickel and cobalt concentrations were much higher in the soft tissues than in the shells (< 0.1 microg g(-1)), this indicates that the shells were not a good indicator for these metals because they do not reflect the environmental bioavailability as do the soft tissues.

Animals↗

Should the patella be resurfaced at total knee replacement?

Total knee replacement (TKR) presumably is replacement of the total knee articular surface. Sometimes it is and sometimes it is not. It is this author's firm conviction that the patella should be resurfaced in the vast majority of cases. Such advocacy must be critically justified and the potential drawbacks and alternatives examined.

Journal Article↗

Metabolic responses of the limpet Patella caerulea (L.) to anoxia and dehydration.

This study examined the metabolic responses of the limpet Patella caerulea (L.) to anoxia and dehydration, attempting to tease apart the effect of these two stressful conditions, which are often not clearly distinguished in experiments. Specimens were exposed to: (a) oxygen-free sea water; (b) oxygen-saturated water (controls); (c) low-humidity air (55% RH); and (d) high-humidity air (100% RH). For each of the treatments, we took samples of five specimens after 6 and 18 h of exposure to the experimental conditions and determined the concentrations in the foot muscle of succinate, acetate, propionate, aspartate and alanine. Exposure to anoxia caused an increase in the levels of succinate (6 and 18 h) and acetate and propionate (18 h) with respect to control specimens. Anoxia also induced a decrease of aspartate and an increase of alanine after both 6 and 18 h. Exposure to both moist and dry air generally had negligible effects on the organic acid levels. Aspartate content increased after 18 h of exposure to moist air. Alanine levels also increased with respect to control values after exposure to air, with dry air having the more pronounced effect. In conclusion, the results of this study suggest that one should be cautious when inferring anaerobic conditions from the simple exposure of intertidal species to air, without strict control of the experimental conditions and actual respiration rates.

Acetates↗

The expression of a caudal homologue in a mollusc, Patella vulgata.

We cloned and analyzed the expression of a caudal homologue (PvuCdx) during the early development of the marine gastropod, Patella vulgata. PvuCdx is expressed at the onset of gastrulation in the ectodermal cells that constitute the posterior edge of the blastopore, as well as in the paired mesentoblasts, the stem cells that generate the posterior mesoderm of the trochophore larva. During larval stages, PvuCdx is expressed in the posterior neurectoderm of the larva, as well as in part of the mesoderm. This is the first report of the expression of a caudal gene in a lophotrochozoan species. The striking similarities with the expression of caudal in other organisms, such as chordates, suggest that a posterior expression of caudal is ancestral to Bilateria.

Amino Acid Sequence↗

A Novel Potassium-Binding Hydrolysis Product of Ascidiacyclamide: A Cyclic Octapeptide Isolated from the Ascidian Lissoclinum patella.

A new octapeptide has been isolated and characterized as its potassium complex from acidic aqueous methanol solutions of ascidiacyclamide, a cyclic peptide isolated from the ascidian Lissoclinum patella. Crystals suitable for X-ray structure determination were orthorhombic, space group P2(1)2(1)2(1), with a = 15.442(3) Å, b = 36.167(1) Å, c = 9.567(3) Å, V = 5343(2) Å(3), Z = 4, and R = 0.069. The structure consists of the macrocyclic cation, three perchlorate anions, and two water molecules. The macrocyclic ring of ascidiacyclamide remains intact, but two oxazoline rings have been opened to form two amino lactones, with the amine protonated. A potassium ion is bound to the N atoms of the thiazole rings (K(1).N(1), 2.47(2) Å; K(1).N(5), 2.50(2) Å) and to the adjacent O (amide) atoms (K(1).O(1), 2.34(1) Å; K(1).O(5), 2.34(1) Å). Two perchlorate anions, located on either side of the plane of the macrocyclic ring, are weakly associated with the potassium cation (K(1).O(10), 2.72(3) Å; K(1).O(16), 2.48(3) Å). This cyclic octapeptide reacts with copper(II) to give a purple solution (lambda(max) 590 nm).

Journal Article↗

New cyclic peptides with cytotoxic activity from the ascidian Lissoclinum patella.

The isolation and structures of a new patellamide (patellamide D) and two new lissoclinamides (lissoclinamides 4 and 5) from the aplousobranch ascidian Lissoclinum patella are described. Structures were determined largely by using two-dimensional NMR techniques and mass spectrometry. These peptides and other members of the patellamide and lissoclinamide families that have been reported previously are found within the obligate algal symbiont of the genus Prochloron. The cytotoxicities of the compounds toward fibroblast and tumor cell lines are reported. One of these compounds, lissoclinamide 4, is markedly more toxic than other members of the family. Structure-activity relationships are discussed.

Animals↗

Structure-activity relationships of the lissoclinamides: cytotoxic cyclic peptides from the ascidian Lissoclinum patella.

Two new lissoclinamides (lissoclinamides 7 and 8) have been isolated from the aplousobranch ascidian Lissoclinum patella. These lissoclinamides are cyclic heptapeptides with the same structural features as lissoclinamides 4 and 5 reported earlier, containing an oxazoline ring, one proline, one valine, two phenylalanine residues, and thiazole and/or thiazoline rings. All four peptides have the same sequence of amino acids around the ring and differ from one another only in their stereochemistry or the number of thiazole and thiazoline rings. The cytotoxicities of the compounds were tested with human fibroblast and bladder carcinoma cell lines and normal lymphocytes. Slight changes in structure resulted in marked differences in the cytotoxicities of these compounds. The most potent is lissoclinamide 7, containing two thiazoline rings, which rivals didemnin B in cytotoxicity in vitro.

Animals↗

The structure of ulithiacyclamide B. Antitumor evaluation of cyclic peptides and macrolides from Lissoclinum patella.

The major cytotoxic metabolites of Lissoclinum patella from Pohnpei have been isolated and identified. Most of the cytotoxicity is attributed to a new cyclic peptide, ulithiacyclamide B [4], which is closely related to ulithiacyclamide [1]. The proposed structure of 4 is based on spectroscopic analysis and chemical degradation. Compound 4 exhibits an IC50 of 17 ng/ml against the KB cell line but does not show selective cytotoxicity against solid tumor cells in the Corbett assay.

Amino Acids↗

Patellamide E: a new cyclic peptide from the ascidian Lissoclinum patella.

A new cyclic peptide, patellamide E [1], was isolated from the ascidian Lissoclinum patella collected at Pulau Salu, Singapore. Its structure was determined by nmr spectroscopy, and its absolute configuration by acid hydrolysis and analysis of the derivatized constituent amino acids by hplc. Patellamide E was mildly cytotoxic against human colon tumor cells in vitro.

Animals↗