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Phosphate mineralization as a skeletal material is uncommon in invertebrate animals and rare in Mollusca. Remarkably, apatite minerals were first reported more than 30 years ago in the periostracum of two species of the mytilid bivalve Lithophaga where shells are mostly constructed of calcium carbonate. This discovery extended the range of biominerals secreted by molluscs but has attracted no subsequent research. In this study we review the occurrence of phosphate mineralization in Lithophaga and putatively allied taxa. Lithophagine bivalves, particularly Lithophaga and the more diverse Leiosolenus species, are well known for their endolithic chemical dissolution of calcareous rocks and corals with calcium-binding lipoproteins secreted by mantle glands. Fluorapatite was identified by X-ray diffraction in an outer layer of the periostracum in six species of Lithophaga. Morphological study by scanning electron microscopy of four species showed the fluorapatite crystals embedded in periostracal material in a layer 10–20 µm thick. Dilute bleach treatment revealed the crystals as densely packed euhedral prisms 250–400 nm in size. The succeeding inner layers of the periostracum were unmineralized. Observations of the developing periostracum of Lithophaga teres suggest that the initial mineralization is in the form of amorphous granules that coalesce and transform into euhedral crystals. Periostracal phosphate was not recorded in other members of the Lithophaginae – Leiosolenus, Botula or Zelithophaga species. Leiosolenus species characteristically have extraperiostracal aragonitic encrustations that can be thick and structurally complex. Published molecular phylogenies of Mytilidae bivalves show a division into two major clades with Lithophaga species in one clade and Leiosolenus species in the other, indicating that the subfamily Lithophaginae as presently understood is polyphyletic. This result implies that the two genera have independent evolutionary pathways to endolithic occupation of calcareous substrates although using similar mantle gland secretions to excavate their crypts. Because fluorapatite is considerably less soluble and harder than calcium carbonate, it is suggested that the phosphate layer of Lithophaga is a functional adaptation to protect their shells from self-dissolution from their rock-dissolving glandular secretions and may also act as defence against other shell-eroding organisms.

New molecular phylogenies of the chemosymbiotic bivalve family Lucinidae, using 18S rRNA, 28S rRNA and cytochrome b genes, include species from genera not previously analysed. Notable additions from Myrteinae are sequences from Rostrilucina, Solelucina and Taylorina species, species of Ustalucina, Gonimyrtea from Leucosphaerinae and additional species of Ctena, Codakia, Lucinoma and Divalucina from Codakiinae. New sequences of Lucininae include the type species of Parvilucina (P. tenuisculpta), Liralucina, Falsolucinoma, Easmithia, Jallenia, Radiolucina and Cardiolucina as well as samples of Loripes orbiculatus from multiple localities. Five major clades, defined as subfamilies, are recognized: Pegophyseminae, Myrteinae, Leucosphaerinae, Codakiinae and Lucininae. Two branches, Fimbriinae and Monitilorinae, are represented by single species. Pegophyseminae are an extremely long-branched group with a sister-group relationship to Leucosphaerinae, while Codakiinae are a sister clade to the Lucininae. In various gene trees, the position of Myrteinae is unstable in relation to Pegophyseminae + Leucosphaerinae, Monitilorinae and Fimbriinae. The Myrteinae are not well resolved, with an ambiguous correlation of molecular and morphological characters. Codakiinae now include Divalucina cumingi, shown to be related to Lucinoma rather than Divaricella and Divalinga of the Lucininae. Leucosphaerinae are a well-supported clade but morphologically disparate, with the positions of Gonimyrtea and Callucina unresolved. Several molecularly distinct subclades are recognized within the Lucininae, especially the Lucinisca, Loripes and Parvilucina groups. Parvilucina species are paraphyletic with P. tenuisculpta, the type species, distinct from the western Atlantic species. Codakia, Ctena and Pegophysema have pan-tropical distributions with former connections disrupted by vicariant events of the closure of the eastern Tethyan and Central American Seaways. Species of Radiolucina, Pleurolucina and Lucinisca are present on either side of the Isthmus of Panama. A new classification of the 96 living lucinid genera is presented, providing a framework for future studies of systematics, ecology, biogeography and bacterial symbioses.

We have modified the periplasmic Escherichia coli glucose/galactose binding protein (GBP) and labelled with environmentally sensitive fluorophores to further explore its potential as a sensor for the evaluation of glucose concentration in airway surface liquid (ASL). We identified E149C/A213R GBP labelled with N,N’-Dimethyl-N-(iodoacetyl)-N’-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)ethylenediamine (IANBD, emission wavelength maximum 536nm) with a Kd for D-glucose of 1.02mM and a fluorescence dynamic range of 5.8. This sensor was specific for D-glucose and exhibited fluorescence stability in experiments for several hours. The use of E149C/A213R GBP-IANBD in the ASL of airway cells grown at air-liquid-interface (ALI) detected an increase in glucose concentration 10 minutes after raising basolateral glucose from 5 to 15mM. This sensor also reported a greater change in ASL glucose concentration in response to increased basolateral glucose in H441 airway cells compared to human bronchial epithelial cells (HBEC) and there was less variability with HBEC data than that of H441 indicating that HBEC more effectively regulate glucose movement into the ASL. The sensor detected glucose in bronchoalveolar lavage fluid (BALf) from diabetic db/db mice but not normoglycaemic wildtype mice, indicating limited sensitivity of the sensor at glucose concentrations

Mitogenomic trees for Bivalvia have proved problematic in the past, but several highly divergent lineages were missing from these analyses and increased representation of these groups may yet improve resolution. Here, we add seven new sequences from the Anomalodesmata and one unidentified semelid species ( Bryopa lata, Euciroa cf. queenslandica, Laternula elliptica, Laternula truncata, Lyonsia norwegica, Myadora brevis, Tropidomya abbreviata, “Abra” sp.). We show that relationships in a mitogenomic tree for the Class are improved by the addition of seven anomalodesmatans from this highly divergent clade, but are still not completely consistent with relationships recovered in studies of nuclear genes. We suggest that some anomalous relationships (for instance the non-monophyly of Bivalvia) may be partially explained by compositional heterogeneity in the mitogenome and suggest that the addition of more taxa may help resolve both this effect and possible instances of long branch attraction. We also identify several curious features about anomalodesmatan mitogenomes. For example, many protein-coding gene boundaries are poorly defined in marine bivalves, but particularly so in anomalodesmatans, primarily due to non-conserved boundary sequences. The use of transcriptomic and genomic data together enabled better definition of gene boundaries, the identification of possible pseudogenes and suggests that most genes are translated monocistronically, which contrasts with many other studies. We also identified a possible case of gene duplication of ND5 in Myadora brevis (Myochamidae) . Mitogenome size in the Anomalodesmata ranges from very small compact molecules, with the smallest for Laternula elliptica (Laternulidae) only 14,622 bp, to Bryopa lata (Clavagellidae) which is at least 31,969 bp long and may be >40,000 bp. Finally, sampled species show a high degree of sequence divergence and variable gene order, although intraspecific variation in Laternula elliptica is very low.

Background and Purpose Hyperglycaemia increases glucose concentrations in airway surface liquid and increases the risk of pulmonary Pseudomonas aeruginosa infection. We determined whether reduction of blood and airway glucose concentrations by the anti-diabetic drug dapagliflozin could reduce P. aeruginosa growth/survival in the lungs of diabetic mice. Experimental Approach The effect of dapagliflozin on blood and airway glucose concentration, the inflammatory response and infection were investigated in C57BL/6J (wild type, WT) or leptin receptor-deficient (db/db) mice, treated orally with dapagliflozin prior to intranasal dosing with LPS or inoculation with P. aeruginosa. Pulmonary glucose transport and fluid absorption were investigated in Wistar rats using the perfused fluid-filled lung technique. Key Results Fasting blood, airway glucose and lactate concentrations were elevated in the db/db mouse lung. LPS challenge increased inflammatory cells in bronchoalveolar lavage fluid from WT and db/db mice with and without dapagliflozin treatment. P. aeruginosa colony-forming units (CFU) were increased in db/db lungs. Pretreatment with dapagliflozin reduced blood and bronchoalveolar lavage glucose concentrations and P. aeruginosa CFU in db/db mice towards those seen in WT. Dapagliflozin had no adverse effects on the inflammatory response in the mouse or pulmonary glucose transport or fluid absorption in the rat lung. Conclusion and Implications Pharmacological lowering of blood glucose with dapagliflozin effectively reduced P. aeruginosa infection in the lungs of diabetic mice and had no adverse pulmonary effects in the rat. Dapagliflozin has potential to reduce the use, or augment the effect, of antimicrobials in the prevention or treatment of pulmonary infection.

Elevation of blood glucose results in increased glucose in the fluid that lines the surface of the airways and this is associated with an increased susceptibility to infection with respiratory pathogens. Infection induces an inflammatory response in the lung, but how this is altered by hyperglycemia and how this affects glucose, lactate and cytokine concentrations in the airway surface liquid is not understood. We used Wild Type (WT) and glucokinase heterozygote (GK+/-) mice to investigate the effect of hyperglycemia, with and without LPS-induced inflammatory responses, on airway glucose, lactate, inflammatory cells and cytokines measured in Bronchoalveolar Lavage Fluid (BALF).

The large, burrowing bivalve Laternula elliptica is an abundant component of shallow-water soft-substrate communities around Antarctica but its congeners are temperate and tropical in distribution and their phylogenetic relationships are obscure. A new molecular analysis of Laternulidae species shows that there are two distinct clades, one of Exolaternula species, E. spengleri and E. liautaudi, possessing a ligamental lithodesma and a larger clade of species lacking the lithodesma. Of the latter, Laternula elliptica is a sister taxon to temperate and tropical species, including those that live around the coasts of Australia from Tasmania to Darwin. It is suggested that L. elliptica was left isolated around Antarctica following the opening of the Tasman Gateway and initiation of the Circum-Antarctic Current as Australia drifted northwards following the final breakup of Gondwana. A further scenario is that as Australia moved closer to Asia, species spread into tropical habitats and more widely to the Red Sea and Japan. Exolaternula species have a likely Tethyan origin and the present-day range is from the Arabian Gulf, around southern Asia and as far north as southern Russia.

We present the first DNA taxonomy publication on abyssal Mollusca from the Clarion-Clipperton Zone (CCZ), central Pacific ocean, using material collected as part of the Abyssal Baseline (ABYSSLINE) environmental survey cruise 'AB01' to the UK Seabed Resources Ltd (UKSRL) polymetallic-nodule exploration area 'UK-1' in the eastern CCZ. This is the third paper in a series to provide regional taxonomic data for a region that is undergoing intense deep-sea mineral exploration for high-grade polymetallic nodules. Taxonomic data are presented for 21 species from 42 records identified by a combination of morphological and genetic data, including molecular phylogenetic analyses. These included 3 heterodont bivalves, 5 protobranch bivalves, 4 pteriomorph bivalves, 1 caudofoveate, 1 monoplacophoran, 1 polyplacophoran, 4 scaphopods and 2 solenogastres. Gastropoda were recovered but will be the subject of a future study. Seven taxa matched published morphological descriptions for species with deep Pacific type localities, and our sequences provide the first genetic data for these taxa. One taxon morphologically matched a known cosmopolitan species but with a type locality in a different ocean basin and was assigned the open nomenclature 'cf' as a precautionary approach in taxon assignments to avoid over-estimating species ranges. One taxon is here described as a new species, Ledella knudseni sp. n. For the remaining 12 taxa, we have determined them to be potentially new species, for which we make the raw data, imagery and vouchers available for future taxonomic study. The Clarion-Clipperton Zone is a region undergoing intense exploration for potential deep-sea mineral extraction. We present these data to facilitate future taxonomic and environmental impact study by making both data and voucher materials available through curated and accessible biological collections.

We report the discovery of highly potent and selective non-steroidal glucocorticoid receptor modulators with PK properties suitable for inhalation. A high throughput screen of the AstraZeneca compound collection identified sulfonamide 3 as a potent non-steroidal glucocorticoid receptor ligand. Further optimization of this lead generated indazoles 30 and 48 that were progressed to characterization in in vivo models. X-ray crystallography was used to gain further insight into the binding mode of selected ligands.

Although species of the chemosymbiotic bivalve family Lucinidae are often diverse and abundant in shallow water habitats such as seagrass beds, new discoveries show that the family is equally speciose at slope and bathyal depths, particularly in the tropics, with records down to 2500 m. New molecular analyses including species from habitats down to 2000 m indicate that these cluster in four of seven recognized subfamilies: Leucosphaerinae, Myrteinae, Codakiinae, and Lucininae, with none of these comprising exclusively deep-water species. Amongst the Leucosphaerinae, Alucinoma, Epidulcina, Dulcina, and Myrtina live mainly at depths greater than 200 m. Most Myrteinae inhabit water depths below 100 m, including Myrtea, Notomyrtea, Gloverina, and Elliptiolucina species. In the Codakinae, only the Lucinoma clade live in deep water; Codakia and Ctena clades are largely restricted to shallow water. Lucininae are the most speciose of the subfamilies but only four species analyzed, Troendleina sp., ‘Epicodakia’ falkandica, Bathyaustriella thionipta, and Cardiolucina quadrata, occur at depths greater than 200 m. Our results indicate that slope and bathyal lucinids have several and independent originations from different clades with a notable increased diversity in Leucosphaerinae and Myrteinae. Some of the deep-water lucinids (e.g. Elliptiolucina, Dulcina, and Gloverina) have morphologies not seen in shallow water species, strongly suggesting speciation and radiation in these environments. By contrast, C. quadrata clusters with a group of shallow water congenors. Although not well investigated, offshore lucinids are usually found at sites of organic enrichment, including sunken vegetation, oxygen minimum zones, hydrocarbon seeps, and sedimented hydrothermal vents. The association of lucinids with hydrocarbon seeps is better understood and has been traced in the fossil record to the late Jurassic with successions of genera recognized; Lucinoma species are particularly prominent from the Oligocene to present day. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 111, 401–420.

Four new species and a new genus of lucinid bivalves are described from shallow and deeper waters in the Indian and West Pacific Oceans. The new genus Scabrilucina (subfamily Lucininae) includes the little-known S. victorialis (Melvill, 1899) from the Arabian Sea and S. vitrea (Deshayes, 1844) from the Andaman Sea as well as a new species S. melvilli from the Torres Strait off northeastern Australia. Ferrocina brunei new species (Lucininae) was recovered from 60 m near oil drilling activities off Borneo; its anatomy confirmed the presence of symbiotic bacteria. Two unusual deeper water species of Leucosphaerinae are described, both species included in on-going molecular analyses; Gonimyrtea ferruginea from 400–650 m in the southwest Pacific and Myrtina reflexa from 200–825 m off Zanzibar and Madagascar.

A new molecular phylogeny of the Lucinidae using 18S and 28S rRNA and cytochrome b genes includes many species from the tropical Western Atlantic as well as additional taxa from the Indo-West Pacific. This study provides a phylogenetic framework for a new taxonomy of tropical Western Atlantic lucinids. The analysis confirmed five major clades—Pegophyseminae, Leucosphaerinae, Myrteinae, Codakiinae and Lucininae, with Monitilorinae and Fimbriinae represented by single species. The Leucosphaerinae are expanded and include Callucina winckworthi and the W. Atlantic Myrtina pristiphora that groups with several Indo-West Pacific Myrtina species. Within the Codakiinae two abundant species of Ctena from the Western Atlantic with similar shells are discriminated as C. orbiculata and C. imbricatula, while in the Indo-West Pacific Ctena bella is a probable species complex. The Lucininae is the most species rich and disparate subfamily with several subclades apparent. Three species of Lucina are recognized in the W. Atlantic L. aurantia, L. pensylvanica and L. roquesana. Pleurolucina groups near to Cavilinga and Lucina, while Lucinisca muricata is more closely related to the E. Pacific L. fenestrata than to the Atlantic L. nassula. A new species of Parvilucina is identified from molecular analyses having been confounded with Parvilucina pectinata but differs in ligament structure. Also, the former Parvilucina clenchi is more distant and assigned to Guyanella.

The systematics of the molluscan class Bivalvia are explored using a 5-gene Sanger-based approach including the largest taxon sampling to date, encompassing 219 ingroup species spanning 93 (or 82%) of the 113 currently accepted bivalve families. This study was designed to populate the bivalve Tree of Life at the family level and to place many genera into a clear phylogenetic context, but also pointing to several major clades where taxonomic work is sorely needed. Despite not recovering monophyly of Bivalvia or Protobranchia—as in most previous Sanger-based approaches to bivalve phylogeny—our study provides increased resolution in many higher-level clades, and supports the monophyly of Autobranchia, Pteriomorphia, Heteroconchia, Palaeoheterodonta, Heterodonta, Archiheterodonta, Euheterodonta, Anomalodesmata, Imparidentia, and Neoheterodontei, in addition to many other lower clades. However, deep nodes within some of these clades, especially Pteriomorphia and Imparidentia, could not be resolved with confidence. In addition, many families are not supported, and several are supported as non-monophyletic, including Malletiidae, Nuculanidae, Yoldiidae, Malleidae, Pteriidae, Arcidae, Propeamussiidae, Iridinidae, Carditidae, Myochamidae, Lyonsiidae, Pandoridae, Montacutidae, Galeommatidae, Tellinidae, Semelidae, Psammobiidae, Donacidae, Mactridae, and Cyrenidae; Veneridae is paraphyletic with respect to Chamidae, although this result appears to be an artifact. The denser sampling however allowed testing specific placement of species, showing, for example, that the unusual Australian Plebidonax deltoides is not a member of Donacidae and instead nests within Psammobiidae, suggesting that major revision of Tellinoidea may be required. We also showed that Cleidothaerus is sister group to the cementing member of Myochamidae, suggesting that Cleidothaeridae may not be a valid family and that cementation in Cleidothaerus and Myochama may have had a single origin. These results highlight the need for an integrative approach including as many genera as possible, and that the monophyly and relationships of many families require detailed reassessment. NGS approaches may be able to resolve the most recalcitrant nodes in the near future.

Pulmonary inflammation disorders represent a major healthcare burden, and novel anti-inflammatory agents are critically needed for the treatment of patients unresponsive to current therapies. In vivo animal models play a key role in the preclinical assessment of novel anti-inflammatory compounds. The implementation of streamlined in vivo experimental designs that are time-and cost-efficient, while keeping animal usage low, is a key consideration for drug optimization programs. The Sephadex rat model of pulmonary inflammation captures many pathophysiologic characteristics of clinical asthma and allergy, such as eosinophilic infiltration andinterstitial edema. Using the in vivo Sephadex model, we compared two different study designs that were implemented to screen and select two novel candidate drugs for a drug discovery project. The traditional one-start design, which utilizes few dose-testing groups with many animals per group, was used to select the first candidate drug. Due to tight timelines, the selection process for the second candidate drug had to be optimized, leading to the development of the novel two-start design, an approach whereby dose ranges are optimized in two experimental phases. Here we show that both study designs were comparable in their generation of robust median effective dose values for selected candidate drugs, as represented by similar confidence interval ratios. However, implementation of the two-start design resulted in approximately 50% fewer animals and 50% less time taken to assess the efficacy of an equal number of compounds compared with the one-start design. These results demonstrate that the two-start design is a more efficient experimental approach, and its widespread implementation in drug optimization programs will impact upon the selection process for candidate drugs with regards to time, cost, and animal usage.

A new molecular phylogeny of the chemosymbiotic bivalve family Lucinidae is presented. Using sequences from the nuclear 18S and 28S rRNA genes and the mitochondrial gene cytochrome b, 105 specimens were analysed representing 87 separate species classified into 47 genera. Samples were collected from a wide range of habitats including mangroves, seagrass beds, shallow sands, offshore muds, and hydrocarbon seeps at depths ranging from the intertidal to over 2000 m. A chronogram, derived from the combined molecular tree, was calibrated using ten lucinid fossils. The trees show five well-supported clades and two single branches of Fimbria fimbriata (Linnaeus, 1758) and Monitilora ramsayi (Smith, 1885). A new classification of Lucinidae is proposed with seven subfamilial divisions: three new subfamilies ‐ Pegophyseminae, Leucosphaerinae, and Monitilorinae ‐ are introduced and Codakiinae, usually treated as a synonym of Lucininae, is revived to include the Lucinoma, Codakia, and Ctena subclades. Membership of the Lucininae and Myrteinae is considerably revised compared with Chavan’s commonly employed ‘Treatise’ classification. Previously considered as a separate family, Fimbriinae is now regarded as a subfamily within Lucinidae. The status of Milthinae is presently equivocal pending further analysis and Divaricellinae is recognized as polyphyletic, and is therefore abandoned, with species and genera now grouped in various places within the Lucininae. Deeper water Lucinidae mainly belong to Leucosphaerinae, Codakiinae (Lucinoma clade), and Myrteinae, with Lucinoma species being most frequently associated with hydrocarbon seeps. Species occurring in seagrass habitats derive largely from Pegophyseminae, Codakiinae, and Lucininae, and species from mangrove habitats derive from the Pegophyseminae and Lucininae.

Rare species of three long-lived lucinid genera, Gibbolucina Cossmann, 1904, Barbierella Chavan, 1938 and Retrolucina n. gen., with origins in the Paleocene and Eocene of western Tethys, are present in the Mozambique Channel area of the southwestern Indian Ocean but absent elsewhere in the Indo-West Pacific. A new species, Gibbolucina zelee n. sp., is described from the Banc de la Zélée and western Madagascar that resembles Miocene species from western France. Since their origin in the Paleocene to the present day Barbierella species have always been rare. New records and images, including syntypes, are provided for Barbierella louisensis (Viader, 1951) from Mauritius and the Mozambique Channel, with Barbierella scitula Oliver & Abou-Zeid, 1986 from the Red Sea regarded as synonym. A new genus, Retrolucina n. gen., is proposed with the living Lucina voorhoevei Deshayes, 1857 (usually called Eomiltha voorhoevei) as type species and also including Lucina defrancei Deshayes, 1857, a strikingly similar species from the Eocene of the Paris Basin. Retrolucina n. gen. differs from Eomiltha Cossmann, 1912 in shape, sculpture and hinge characters. Monitilora Iredale, 1930, another genus of Paleocene or earlier origins, includes a few living species in the Indo-West Pacific and is now identified from Mozambique with Monitilora sepes (Barnard, 1964) (formerly Phacoides sepes Barnard, 1964). It is suggested that Gibbolucina, Barbierella and Retrolucina n. gen. species became isolated in the western Indian Ocean following the closure of the Tethyan Seaway in the early Miocene while their congeners in western Tethys became extinct. The survival of these rare genera, with restricted geographical ranges and seemingly small populations, runs counter to current ideas concerning long-term extinction risk., Des espèces rares appartenant à trois genres anciens de lucines, Gibbolucina Cossmann, 1904, Barbierella Chavan, 1938 et Retrolucina n. gen., dont l'origine se situe au Paléocène et à l'Éocène en Téthys occidentale, sont présentes dans la zone du Canal du Mozambique dans le sud-ouest de l'océan Indien, et absentes ailleurs dans l'Indo-Ouest Pacifique. Une nouvelle espèce, Gibbolucina zelee n. sp., est décrite du Banc de la Zélée et de l'ouest de Madagascar; elle ressemble à une espèce du Miocène de l'ouest de la France. Depuis leur origine au Paléocène jusqu'à nos jours, les espèces de Barbierella ont toujours été rares. Des signalisations nouvelles et des illustrations, dont celles de syntypes, sont données pour Barbierella louisensis (Viader, 1951) de l'île Maurice et du Canal du Mozambique, et Barbierella scitula Oliver & Abou-Zeid, 1986 de la Mer Rouge est donnée comme synonyme. Le nouveau genre Retrolucina n. gen. est établi avec pour espèce type l'espèce actuelle Lucina voorhoevei Deshayes, 1857 (habituellement appelée Eomiltha voorhoevei), et comprenant également Lucina defrancei Deshayes, 1857, une espèce de l'Éocène du Bassin Parisien qui lui est étonnamment semblable. Retrolucina n. gen. diffère d'Eomiltha Cossmann, 1912 par sa forme, sa sculpture et les caractères de sa charnière. Monitilora sepes (Barnard, 1964), du Mozambique (auparavant Phacoides sepes Barnard, 1964), est classée dans le genre Monitilora Iredale, 1930, un autre genre dont l'origine remonte au moins au Paléocène et qui comprend un petit nombre d'espèces actuelles de l'Indo-Ouest Pacifique. Les espèces de Gibbolucina, Barbierella et Retrolucina n. gen. ont pu se retrouver isolées dans l'Ouest de l'océan Indien à la suite de la fermeture de la Téthys au début du Miocène alors que leurs congénères de la Téthys occidentale se sont éteintes. La survie de ces genres rares, avec des aires de distribution restreintes et apparemment de petites populations, va à l'encontre des idées classiques sur les risques d'extinction à long terme.

A scanning electron microscope study of the periostracum of 50 species of venerid bivalves revealed that periostracal calcification in the form of aragonitic needles and shorter pins is widespread within the family. Together with organic and sediment coatings that are found in some species, these needles form an integral part of the functional shell. Visible as a white ‘crust’ on the outside of shells, long slender needles (up to 400 mm long and 1 mm wide) without adherent material are seen in species of Tivela and Lioconcha and in Gomphina undulosa. Other venerids including Pitar species, Mysia undata and Compsomyax subdiaphana have short pins, capped with a fibrous organic matrix and significant coatings of sediment. Callocardia hungerfordi and Clementia papyracea have very thick sediment coatings underlain by short pins, while Gafrarium and Circe species have short pins with a thin, robust, organic coating and little particulate material. Finally, there are species, including Venus verrucosa, Chione elevata and Mercenaria mercenaria, where minute, ,1 mm long pins also underpin a thin organic coating. Details of formation were studied in Tivela lamyi and Lioconcha ornata, where the needles are elongate hexagonal crystals of aragonite enveloped by an organic sheath, which grow at their proximal ends from within the periostracum, connected to the outer mantle epithelium via narrow channels. Growth of needles ceases following the onset of shell calcification. The distribution of the periostracal structures was examined in relation to a published molecular phylogeny that recognized two major clades within the family. Larger needles and pins are confined to the clade that includes subfamilies Pitarinae, Gouldinae, Meretricinae and Petricolinae, while submicron-sized pins are found only in the Venerinae and Chioninae of the second clade. Calcified periostracal structures appear to be absent in Tapetinae and Dosiniinae.

The structure of the ctenidia of the Indo-West Pacific chemosymbiotic lucinid bivalve Anodontia (Euanodontia) ovum was investigated by electron microscopy. Ctenidial filaments are similar in general morphology to those described from other Lucinidae, with a ciliated zone, a short intermediary zone and a thick abfrontal zone composed largely of bacteriocytes separated by narrow intercalary cells. The bacteriocyte zones of adjacent filaments are fused in the distal part to form short cylindrical channels. The apices of intercalary cells project as cytoplasmic protrusions in the form of spiky tufts, with sheets and tendrils spreading over adjacent bacteriocytes. Compared with other lucinids A. ovum lacks abfrontal granule cells, mucocytes are infrequent and the bacteriocyte channels are short. Three morphotypes of symbiotic bacteria were detected, associated with the bacteriocyte zone of the ctenidial filaments: (1) all bacteriocytes contained abundant bacteria 3-5 mu m long and 0.5-1.0 mu m wide, enclosed in single vacuoles; (2) some bacteriocytes possessed spherical vesicles enclosing masses of smaller rod-shaped bacteria c. 1.0 mu m long; (3) probable spirochaete bacteria, 8-10 mu m long and 0.3 mu m wide, were abundant within the apical cytoplasmic protrusions of the intercalary cells. Preliminary molecular analysis using 16S rRNA gene sequences has so far identified only one bacterial symbiont, from the gamma division of Proteobacteria grouping in a cluster of symbiotic thiotrophs. This symbiont of A. ovum is closely similar to a symbiont previously reported from the western Atlantic lucinid Anodontia schrammi (originally cited as A. philippiana).

The giant bivalve Lucina megameris Dall, 1901, from the late Eocene White Limestone Group of Jamaica and by far the largest known species of the family Lucinidae, is placed in a new genus Superlucina. Apart from its large size, with a shell height exceeding 310 mm, it is distinguished from other genera, such as Pseudomiltha and Eomiltha by external shell characters and the extremely long and narrow, anterior adductor muscle scar. Features preserved on internal moulds suggest that, in common with living Lucinidae, S. megameris was chemosymbiotic with sulphide - oxidizing bacteria housed in the gills. Palaeoenvironmental evidence suggests a habitat in oligotrophic, shallow waters, probably in seagrass beds, with an associated molluscan fauna includ- ing large cardiids that may have been photosymbiotic. Super- lucina is considerably larger than any living lucinid that range in size from 3 to 150 mm with most encompassed within 5-30 mm. From the Jurassic onwards, a few other large lucinids are known from cold seep sites, with several other records from possible shallow water seagrass beds.

Chronic obstructive pulmonary disease (COPD) exacerbations are a major cause of morbidity and mortality in COPD. There are several current treatments that are used today, but still a large unmet medical need. We discuss current treatments for COPD exacerbations and also allude to potential new routes to therapy with reference to the known pathobiology of a COPD exacerbation.

A new molecular phylogeny is presented for the highly diverse, bivalve molluscan subclass Heterodonta. The study, the most comprehensive for heterodonts to date, used new sequences of 18S and 28S rRNA genes for 103 species from 49 family groups with species of Palaeoheterodonta (Trigoniidae, Margaritiferidae and Unionidae) as outgroups. Results confirm previous analyses that the Carditidae/Astartidae/Crassatellidae clade is basal to all other heterodonts including Anomalodesmata (often classified as a separate subclass or order). Thyasiroidea occupy a near basal position between the Crassatelloidea and Anomalodesmata. Lucinidae form a well-supported monophyletic group distinct from Thyasiridae and Ungulinidae. The Solenoidea and Hiatelloidea link as sister groups distant from the Tellinoidea and Myoidea, respectively, where they had been previously associated. The position of the Gastrochaenidae is unstable but does not group with myoidean taxa. Species of four families of Galeommatoidea form a clade that also includes Sportellidae of the Cyamioidea. The Cardioidea and Tellinoidea form highly supported, long branched, individual clades but group as sister taxa. A major clade including Veneroidea, Mactroidea, Myoidea and other families is given the unranked name Neoheterodontei. There is no support for a separate order Myoida (Myoidea and Pholadoidea). Dreissenidae group within the clade including Myidae, Corbulidae, Pholadidae and Teredinidae. The Corbiculoidea is confirmed as polyphyletic with the Sphaeriidae and Corbiculidae forming separate clades within the Neoheterodontei; Corbiculidae grouping with the Glauconomidae. Hemidonacidae are unrelated to the Cardiidae, as previously proposed, but nest within the Neoheterodontei. The Gaimardiidae group near to the Ungulinidae and not with Cyamioidea where most recently classified. The family Ungulinidae, previously classified in the Lucinoidea, forms a well-supported clade within the Neoheterodontei and is elevated to superfamily rank — Ungulinoidea. The monophyletic status of Glossoidea, Arcticoidea and Veneroidea is unconfirmed. A brief review of the fossil record of the heterodonts indicates that the basal clades of Crassatelloidea, Anomalodesmata and Lucinoidea diverged very early in the Lower Palaeozoic. Other groups such as the Hiatelloidea, Solenoidea, Gastrochaenidae probably were of late Palaeozoic origins. The Cardioidea and Tellinoidea originated in the Triassic while major groups of Neoheterodontei radiated in the Late Mesozoic. The phylogenetic position of the Thyasiroidea and Galeommatoidea suggests a longer fossil history than has so far been recognized.

Molecular analyses of 13 species of the marine bivalve family Thyasiridae, using sequences from 18S rRNA and 28S rRNA genes, showed that the family is monophyletic despite the anatomical disparity and inclusion of both chemosymbiotic and asymbiotic species. This new analysis also confirmed that the three families (Thyasiridae, Lucinidae and Ungulinidae), previously included in the Lucinoidea, were not closely related. Four species of Ungulinidae grouped within a clade containing Veneridae, Arcticidae and Mactridae. In relation to a range of other heterodont bivalves, Thyasiridae occupied a near basal position, apart from a clade comprising Carditidae/Astartidae/Crassatellidae. The earliest thyasirid recognized in the fossil record is a species from the Lower Cretaceous of England. Within the Thyasiridae, some groups can be identified but relations between these are weakly supported. Amongst the taxa analysed, those with symbiotic bacteria and two ctenidial demibranchs belong to at least three groups, while there is some support for a clade of asymbiotic taxa with single demibranchs. In recognition of the monophyletic status of the Thyasiridae, distinct from all other heterodont bivalves, we elevate the rank to superfamily Thyasiroidea.

Glucose concentrations in lung airway surface liquid are normally lower than those of plasma (0.4mM vs 5mM). However, glucose concentration increases (up to 4mM) during hyperglycaemia, which would make the lung more susceptible to colonisation by respiratory pathogens. We investigated how glucose and lactate concentrations were modified by hyperglycaemia and inflammation using two mouse models; the leptin deficient db/db (n=10-12) and the glucokinase deficient GK -/- mouse (n=6) ± intranasal lipopolysaccharide (LPS, 0.0875mg/g) challenge. Blood glucose was increased in GK (15.1±0.6mM) compared to wild type (9.4±0.2mM, p 6 /ml, p 6 /ml) and db/db mice (0.29±0.04x10 6 /ml). BAL lactate was concomitantly increased to 0.16±0.01mM, but BAL glucose was significantly depleted at 0.04±0.01mM. These data indicate that elevation of blood glucose increases glucose and lactate in the lung and provide evidence that alveolar macrophages utilise glucose and further elevate lactate during acute inflammation.

Recent molecular analyses have demonstrated that the traditional Lucinoidea, comprising the extant families Lucinidae, Thyasiridae, Ungulinidae, Fimbriidae, and Cyrenoididae, is not monophyletic. Thyasiridae and Ungulinidae are unrelated to Lucinidae, a result corroborated by clear morphological differences between the groups. Chemosymbiosis in Thyasiridae and Lucinidae has been independently derived. Within the family Lucinidae, previous ideas of relationship and subfamilial divisions based on shell characters have little support from molecular results. Anatomical characters of the ctenidia, mantle gills, and posterior apertures have potential in phylogenetic analysis but rigorous analysis of shell characters is also needed. Although there is a good fossil record of Lucinidae throughout the Cenozoic and Mesozoic, in the Palaeozoic fossils are less frequent and most need reappraisal. The Silurian Ilionia prisca is probably the earliest fossil with convincing lucinid features, followed in the Devonian by Phenacocyclas and some Paracyclas species. © 2006 The Linnean Society of London, Zoological Journal of the Linnean Society, 2006, 148, 421–438.

OBJECTIVE To establish the equivalence between the tension-free vaginal tape (TVT) and the suprapubic urethral support sling (SPARC). Approximately 35% of women have stress urinary incontinence (SUI), and although TVT is now perceived as the standard treatment, the SPARC is a very similar procedure and is thought to have fewer peri-operative complications. PATIENTS AND METHODS Patients with clinical SUI were recruited from public and private urology/urogynaecology clinics, and participated in the trial of TVT vs SPARC. The primary outcome was bladder perforation; secondary outcomes were blood loss, voiding difficulty, urgency, and cure of SUI symptoms. Sample size calculations, based on an estimated 2% perforation rate, showed that 290 patients would be needed to detect a clinically significant difference of 5%. Stratification was by previous incontinence surgery and the experience of the surgeon. RESULTS There were 301 operations; the difference in bladder perforations was not statistically significant, at one/147 TVT (0.7%), and three/154 SPARC (1.9%), with the difference in rate of 0.013 (95% confidence interval (CI) − 0.01 to 0.04; odds ratio 2.89, 95% CI 0.30–28.21; P = 0.62), and nor were differences in estimated blood loss of >100 mL (TVT, 32/147, 21.8%; SPARC 28/154, 18.2%); de novo urgency (TVT 15/37, 40.5%; SPARC 14/33, 42.4%), objective cure (TVT 143/147, 97.3%; SPARC 148/152, 97.4%) or vaginal mesh erosion (TVT 7/147, 4.8%; SPARC 16/152, 10.5%). Acute urinary retention (TVT none of 147; SPARC 10/154, 6.5%; odds ratio ∞, 95% CI 2.2–∞; P = 0.002) and subjective cure (TVT 128/147, 87.1%; SPARC 117/153, 76.5%; odds ratio 2.07, 95% CI 1.13–3.81; P = 0.03) were statistically significantly different. CONCLUSION These results are consistent with clinical equivalence between TVT and SPARC for bladder perforation. There was no statistically significant difference between TVT and SPARC in blood loss, urgency or objective cure of SUI symptoms at 6 weeks. However, SPARC was more difficult to adjust correctly, and a statistically significant number of patients required loosening of the tape in theatre (P = 0.002). TVT had a lower rate of vaginal erosion and a statistically significantly higher cure rate of subjective SUI symptoms than SPARC. Overall, voiding difficulty (loosening of the tape), urgency and vaginal mesh erosion were the most important clinical problems. This trial showed the importance of testing new devices which appear to be similar, but which might have relevant differences. There was no financial assistance for this study, and a long-term follow up is planned.

The bivalve superfamily Lucinoidea is usually considered to comprise six separate families: Lucinidae, Thyasiridae, Ungulinidae, Fimbriidae, Mactromyidae and Cyrenoididae. Chemoautotrophic chemosymbiosis with sulphide-oxidizing bacteria is present in all studied species of Lucinidae, Fimbriidae and many, but not all, Thyasiridae. However, it is absent from Ungulinidae. The Mactromyidae are likely to be an entirely fossil group with doubtful affinities with Lucinoidea. The Cyrenoididae are poorly investigated, but anatomical features suggest they are unrelated to lucinoids. To investigate phylogenetic relationships within the Lucinoidea and test hypotheses concerning the evolution of the chemosymbiosis, a molecular study was made using sequences of 18S and 28S rRNA genes. The study incorporated species of Ungulinidae (two genera, two species), Thyasiridae (three species), Fimbriidae (one species) and many Lucinidae (31 species, 19 genera) as well as a range of outgroups representing major groups of heterodont and palaeoheterodont bivalves. The results demonstrate that the monophyly of the Lucinoidea is not supported. The Ungulinidae and Thyasiridae are unrelated to the Lucinidae. Ungulina and Diplodonta of the Ungulinidae group with a clade comprising Veneroidea, Arcticoidea and Mactroidea. The three Thyasira species analysed form a monophyletic branch in a basal position among the heterodont bivalves. The only member of the Fimbriidae examined, Fimbria fimbriata, groups within the Lucinidae and separation as a family is not supported. The Lucinidae form a monophyletic group within which several distinct and well-supported clades and lineages are recognized: the Myrtea clade, the ‘Anodontia’ clade, Fimbria lineage, Phacoides pectinatus lineage, and two clades comprising all other lucinids. The implication of non-monophyly of the superfamily Lucinoidea is that Thyasiridae represent an independent acquisition of bacterial chemosymbiosis and this is reflected in major morphological differences from the Lucinidae.

Fractal or focal dimension (FD) analysis is a valuable tool to identify physiologic stimuli at the cellular and tissue levels that allows for quantification of cell perimeter complexity. The FD analysis was determined on fluorescence images of caffeine- or epinephrine-treated (or untreated control) killifish Fundulus heteroclitus (Linneaus) melanophores in culture. Cell perimeters were indicated by rhodamine-phalloidin labeling of cortical microfilaments using box-counting FD analysis. Caffeine-treated melanophores displayed dispersed melanosomes in cells with less serrated edges and reduced FD and complexity. Complexity in epinephrine-treated cells was significantly higher than the caffeine-treated cells or in the control. Cytoarchitectural variability of the cell perimeter is expected because cells change shape when cued with agents. Epinephrine-treated melanophores demonstrated aggregated melanosomes in cells with more serrated edges, significantly higher FD and thus complexity. Melanophores not treated with caffeine or epinephrine produced variable distributions of melanosomes and resulted in cells with variably serrated edges and intermediate FD with a larger SE of the regression and greater range of complexity. Dispersion of melanosomes occurs with rearrangements of the cytoskeleton to accommodate centrifugal distribution of melanosomes throughout the cell and to the periphery. The loading of melanosomes onto cortical microfilaments may provide a less complex cell contour, with the even distribution of the cytoskeleton and melanosomes. Aggregation of melanosomes occurs with rearrangements of the cytoskeleton to accommodate centripetal distribution of melanosomes. The aggregation of melanosomes may contribute to centripetal retraction of the cytoskeleton and plasma membrane. The FD analysis is, therefore, a convenient method to measure contrasting morphologic changes within stimulated cells.

Graft thrombosis is an important cause of early (4 weeks) renal graft loss. Reports show that heparin reduces the incidence of early renal allograft thrombosis. Routine peri-operative administration of unfractionated heparin was introduced in our unit in 1994. We conducted a retrospective study of 254 transplants, undertaken in children, between 1987 and 2000. There were 126 children who did not receive heparin (group 1) and 128 who did (group 2). Recipient characteristics and immunosuppression were similar in both groups. The incidence of graft loss secondary to thrombosis was compared between the groups. Variables previously identified with increased risk of graft loss, including donor age, recipient age, cold ischaemia time (CIT), multiple donor vessels, surgical complications, and side of graft donation, were examined using logistic regression. Thrombosis occurred in 14 grafts in group 1 and 11 grafts in group 2 (odds ratio 0.7, 95% confidence interval 0.3-1.6, P=not significant). The mean time to graft loss was similar in groups 1 and 2 (6.6, SD 3.9, range 2-12 days and 7.9, SD 4.4, range 1-14 days, respectively) ( P=0.445). Young recipient age ( P=0.006), young donor age ( P=0.009), increasing CIT ( P=0.007), and surgical complications ( P=0.002) increased the risk of graft thrombosis. A reduction in the incidence of early renal allograft thrombosis upon introduction of heparin was not demonstrated.

Many specialist obstetricians and gynaecologists feel inadequately trained to deal with certain situations such as injury to bowel, bladder, ureter and major vessels, and value further training to prevent and manage these problems. We present the structure, objectives and rationale for a surgical skills workshop, which is an intensive practical learning experience aimed to improve the performance of obstetricians and gynaecologists. The overall objective of the workshop is improvement in the prevention and management of complications in obstetric and gynaecological surgery. Over 100 participants have completed the workshop so far. Pre-workshop preparation includes anatomical illustrations to guide reading and a training video describing surgical skills, ewe anatomy and hysterectomy in the ewe. There are four modules: anatomy includes an interactive lecture, cadaveric dissection and examination of prosections with specific learning tasks. Surgical skills involves the demonstration, practice and learning of techniques needed to deal with unexpected operative injury to bowel, bladder, ureter and major blood vessels. Live animal surgery on a ewe allows further supervised practice of the previously learned skills plus the repair of serious vascular injury. Case presentations allow each participant to present a complicated case in a facilitated group session with discussion and feedback from their peers. This session is controlled, non-threatening and a valuable interactive learning experience. Participant feedback suggests that this workshop format is useful and appears to improve the confidence, competence and performance of the participants. This workshop is presented as a template on which other educational activities can be developed.

Bivalves are an ancient and ubiquitous group of aquatic invertebrates with an estimated 10 000–20 000 living species. They are economically significant as a human food source, and ecologically important given their biomass and effects on communities. Their phylogenetic relationships have been studied for decades, and their unparalleled fossil record extends from the Cambrian to the Recent. Nevertheless, a robustly supported phylogeny of the deepest nodes, needed to fully exploit the bivalves as a model for testing macroevolutionary theories, is lacking. Here, we present the first phylogenomic approach for this important group of molluscs, including novel transcriptomic data for 31 bivalves obtained through an RNA-seq approach, and analyse these data with published genomes and transcriptomes of other bivalves plus outgroups. Our results provide a well-resolved, robust phylogenetic backbone for Bivalvia with all major lineages delineated, addressing long-standing questions about the monophyly of Protobranchia and Heterodonta, and resolving the position of particular groups such as Palaeoheterodonta, Archiheterodonta and Anomalodesmata. This now fully resolved backbone demonstrates that genomic approaches using hundreds of genes are feasible for resolving phylogenetic questions in bivalves and other animals.

Pigmentary organelle translocations within fish chromatophores undergo physiological color changes when exposed to external signals. Chromatophores can be isolated in high yields, and their pigmentary organelles can be tracked readily by microscopy. The combined efforts of morphology and biomolecular chemistry have led to the identification of and determination of the interrelationships between cytoskeletal elements and accessory proteins, motor molecules, cytomatrix, and pigmentary organelles of various sizes. Fish chromatophores have been classified as fast, intermediate, and slow translocators, based on the relative numbers of microtubules. Studies on cultured goldfish (Carassius auratus L.) xanthophores for over 20 years have demonstrated that in this slow translocator, tubulovesicular structures of the smooth endoplasmic reticular (SER) cisternae are involved in the disperson and aggregation of associated carotenoid droplets (CD) with some involvement of cytoskeletal elements. Killifish (Fundulus heteroclitus L.) melanophore, a fast translocator, was also examined. Recent work demonstrates a bright fluorescent "starburst"-like spot that we call an actin filament-organizing center (AFOC) with radiating microfilaments, akin to the microtubule-organizing center (MTOC) with radiating microtubules. Melanosomes translocate single-file on microtubules and are not associated with SER cisternae. Slower CD dispersion or aggregation in goldfish xanthophores seems to be predominantly microfilament-based transport, or microfilament- and microtubule-based transport, respectively. Faster melanosome translocations in killifish melanophores are based on microtubules, with our evidence indicating microfilament involvement. Neural crest-derived chromatophores are models for vesicular transport in axons, and immunocytochemical and imaging technologies may help to elucidate the cellular transport mechanisms.

This taxonomic revision concerns the Australian and Indo-Pacific species of small lucinid bivalves possessing a deeply inset internal ligament. Nine species of Pillucina are recognised of which four occur around Australia including the two new species, P. pacifica and P. australis. Two other new species are described; P. denticula from South Africa and P. mauritiana from Mauritius. Pillucina vietnamica is common along the Queensland coast and P. symbolica the only species previously recorded from Australia is considered conspecific with the wide ranging species P. pisidium. A new genus, Chavania, includes two species; C. striata is widely distributed in the Indo-West Pacific including eastern and western Australia, while C. erythraea is restricted to the Arabian Peninsula. Two species of Wallucina live around Australia, W. assimilis is endemic occurring at southerly locations from New South Wales to North West Cape, while the tropical species, W. fijiensis, is found in island habitats of Queensland but is widely distributed in the Indo-West Pacific. Another new genus, Funafutia, is proposed for the species, F. levukana, recorded from Australia for the first time. Details of anatomy are provided for Pillucina vietnamica, W. assimilis and C. striata. Symbiotic bacteria are confirmed for the first time in the lateral zone of gill filaments of Pillucina vietnamica and Wallucina assimilis.

Four main types of radula are found within the gastropod superfamily Conoidea. Type 1, the Clavidae type has five teeth in each row with comb-like laterals and flat pointed marginals. Type 2, the Turridae type has two or three teeth in a row with the marginals comprised of teeth with the duplex or wishbone form. Type 3, the Pseudomelatomidae/Pervicaciinae type has two or three teeth in a row with the marginals curved and solid. Type 4, the hypodermic type has two hollow, enrolled, marginal teeth in each row with the radular membrane reduced or absent. Study of the maturing teeth along the radular membrane in gastropods with the Type 2 radula shows that the duplex teeth are not formed from two separate elements but develop from a flat plate, by thickening of the tooth edges and elevation of the posterior edge from the membrane. Semi-enrolled and enrolled teeth of Pilsbryspira, Imaclava and Toxiclionella develop in a similar way. In Conidae where a vestigial radular membrane is attached only at the base of the teeth, the teeth are enrolled from first formation in the radular sac. This study demonstrates that duplex and enrolled teeth develop by a similar process, allowing the possibility of deriving hypodermic teeth from duplex clades as has been suggested by previous phylogenetic analysis.

Recent years have seen a growing interest in the direct and indirect roles of organisms, and more explicitly of organic matter, in the crystallization of minerals in sediments. Two bivalve genera form extensive biofilms that are apparently responsible for the growth of a variety of isolated crystals together with a marine cement. Granicorium indutum and Samarangia quadrangularis are infaunal species that cover their shells with a cemented coating of sand, sculpted to mimic the surface ornament typical of many bivalves. Granicorium has an exceptionally mobile mantle margin that produces large volumes of mucus, harboring a diverse microbial community. The sand grains surrounding both species are initially bound by a biofilm that provides structural integrity but also acts, like others, as a template for the crystallization of a variety of carbonate polymorphs. These include varied prismatic crystals, rice-grain and wheatsheaf forms and, more importantly, large volumes of acicular crystals indistinguishable from typical marine cements. The distribution of these crystals is related to positions accessible to the mantle of the animal and in Samarangia appears to result from the active emplacement of grains and mucus several times a year. Summing the evidence, it seems that these species are responsible not only for the biomineralization involved in the formation of their shells but also for crystallization mediated through biofilms and for the generation of cements that are morphologically indistinguishable from typical marine cements. The boundaries between these three strategies may be closer than we think.

Field observations of Oliva tigridella in Queensland revealed a diet of small gastropods, bivalves, echinoids and holothurians. Foraging animals were observed carrying prey attached to the rear of the foot. Up to seven prey items were recorded on the foot. Analysis of gut co ntents showed a high frequency of holothurian spicules although these were rare amongst items found on the foot. This study reveals a previously unrecorded catholic diet of olives although small gastropods and bivalves are important components.

Two veneroidean bivalves Granicorium indutum from Australia and Samarangia quadrangularis from the tropical Indo-Pacific region, cement a thick, hard layer of sand over most of their shells. In Granicorium this layer forms low commarginal ribs while in Samarangia it forms more prominent radial features. Sand grains are cemented to the shell and to each other with growths of a crystalline aragonitic cement similar in morphology to inorganic marine cements. Both species secrete mucus layers at the growing shell margin which initially hold the sediment grains together and form a substrate for the nucleation and growth of calcium carbonate crystals. The ribs of Samarangia are formed by the accretion of successive sheets of spherulitic growths. In G. indutum, the middle and outermost of two inner mantle folds are large, glandular and capable of considerable extension beyond the shell margin. Mucus secreted by the folds contains abundant bacteria and small calcium carbonate crystals. It is proposed that initial nucleation of the calcium carbonate cement takes place within this biofilm possibly mediated by the bacteria. The function of the sand layers is unknown but predation resistance and protection of the shells from endobionts are the most likely possibilities.

The fine structure of the salivary glands of Cymatium intermedius is described. It is concluded that both the posterior acid secreting and anterior acinous lobes are homologous with the salivary glands of other prosobranchs. At least six different types of secretion have been identified and their possible roles discussed. The anterior lobes secrete mucin and proteins believed to be enzymes, and the posterior lobes are specialized for acid production as well as secreting protein (probably a peptide toxin). The mechanisms of acid secretion and protection from damage to the snail before its release are also considered.

BACKGROUND PSP94 (prostate secretory protein of 94 aa; also called PIP), one of the predominant proteins secreted into the seminal fluid, was proposed as an independent diagnostic/prognostic marker for prostate cancers. It was also shown to inhibit rat prostate cancer growth. In this study, we investigated the effect of purified PSP94 on the growth of androgen-independent human prostate cancer cells (PC3) and its potential mechanism of action. METHODS AND RESULTS PSP94, in a dose- and time-dependent manner, inhibited the growth of PC3 cells. The protein demonstrated a stronger inhibitory effect on the colony-forming ability of PC3 cells in soft agar. A daily injection of PSP94 at 5 μg/kg/body weight resulted in a 50–60% inhibition in the growth of PC3 xenografts in athymic mice. PC3 cell growth inhibition by PSP94 resulted from cell death characteristic of morphological apoptosis, which was confirmed by dual fluorescence microscopy, electron microscopy, and DNA fragmentation assays. Mechanistic studies indicated that PSP94 enhanced the expression of proapoptotic protein Bax without affecting Bcl-2 levels. CONCLUSIONS This study suggests that PSP94 may represent a novel, apoptosis-based, antitumor agent applicable to the treatment of hormone-refractory human prostate cancers. Prostate 38:118–125, 1999. © 1999 Wiley-Liss, Inc.