Chemical and boron isotopic compositions of volcanics from ODP Sites 135-834 to 135-840

The influence of fluid flux on petrogenesis in the Tonga-Kermadec Arc was investigated using ion microprobe measurements of B/Be and boron isotope ratios (11B/10B) to document the source and relative volumes of the fluids released from the subducting oceanic plate. We analyzed young lavas from eight different islands along the Tonga-Kermadec Arc, as well as glass shards in volcanic sediments from Ocean Drilling Program (ODP) Site 840, which record the variations in the chemistry of Tonga magmatism since 7 Ma. B/Be is variable (5.8-122), in young Tonga-Kermadec Arc lavas. In contrast, glass shards from around 3 to 4 Ma old volcanic sediments at Site 840 have the highest B/Be values yet reported for arc lavas (18-607). These values are too high to be related simply to a sediment influence on petrogenesis. Together with very high d11B values (-11.6 to +37.5) for the same shards and lavas these data indicate that most of the B is derived from fluid escaped from the subducting altered Pacific oceanic crust, rather than from sediment. High d11B values also reflect large degrees of isotopic fractionation in this cold fast subduction zone. Lower d11B values noted in the Kermadec Arc (17 to -4.4) are related to the influence of sediment eroded from New Zealand and slower convergence. High fluid flux (B/Be) is synchronous in Tonga and the Marianas at 3 to 4 Ma and may be related to acceleration of the Pacific Plate just prior to this time. The timing of maximum B/Be at 3 to 4 Ma correlates with maximum light rare earth (LREE) and high field strength element depletion. This suggests maximum degrees of partial melting at this time. Although thinning of the arc lithosphere during rifting to form the Lau Basin is expected to influence the arc geochemistry, variable aqueous fluid flux from the subducting plate alone appears capable of explaining boron and other trace element systematics in the Tonga-Kermadec Arc with no indication of slab melting.

Geochemistry of ODP Leg 135 vitrophyric rhyolite samples

In-situ proton-microprobe analyses are presented for glasses, plagioclases, pyroxenes, olivines, and spinels in eleven samples from Sites 834-836, 839, and 841 (vitrophyric rhyolite), plus a Tongan dacite. Elements analyzed are Mn, Ni, Cu, Zn, Ga, Rb, Sr, Y, Zr, Pb, and Sn (in spinels only). The data are used to calculate two sets of partition coefficients, one set based on the ratio of element in mineral/element in coexisting glass. The second set of coefficients, thought to be more robust, is corrected by application of the Rayleigh fractionation equations, which requires additional use of modal data. Data are presented for phenocryst core-rim phases and microphenocryst-groundmass phases from a few samples. Comparison with published coefficients reveals an overall consistency with those presented here, but with some notable anomalies. Examples are relatively high Zr values for pyroxenes and abnormally low Mn values in olivines and clinopyroxenes from Site 839 lavas. Some anomalies may reflect kinetic effects, but interpretation of the coefficients is complicated, especially in olivines from Sites 836 and 839, by possible crystal-liquid disequilibrium resulting from mixing processes.

Late Neogene biostratigraphy of ODP Leg 135 sites

Integration of biostratigraphic and magnetostratigraphic results from Leg 135 sites has given additional information as to the position and reliability of various bioevents compared with previously published results. Two sites (834: Gilbert to Brunhes; and 836: Brunhes) provided excellent magnetic and biostratigraphic data. From these it is suggested that some bioevents are older than previously recorded: the first appearances (FAs) of Emiliania huxleyi (within the Brunhes Chron, at the same level as the FA of Helicosphaera inversa) and Globorotalia (Truncorotalia) truncatulinoides (within the upper Gauss Chron), and the last appearance (LA) of Gr. (Tr.) tosaensis (upper Matuyama Chron). The FA of Gr. (Tr.) crassaformis hessi is variable, but the oldest occurrence is just below the Cobb Mountain Subchron. Other key bioevents, such as the LAs of Discoaster pentaradiatus (just above the Réunion Subchron), D. tamalis (within the lower reversed part of the Matuyama Chron), Sphenolithus (lower Gauss Chron), and Amaurolithus primus (topmost Gilbert Chron) appear higher than previously recorded. Some key biostratigraphic taxa, such as Globigerinoides quadrilobatus fistulosus, Pulleniatina finalis, P. primalis, and Sphaeroidinella dehiscens, are either rare or their distribution is sporadic to the extent that they are unsuitable for biostratigraphic use in the area studied. Because of the rarity of P. primalis, the FA of Globorotalia (Globorotalia) multicamerata has been used to mark the base of Zone N17B. Though levels are present at most sites in which populations of Pulleniatina are sinistrally coiled, it is difficult to equate these coiling changes with previous records.

Miocene to Pleistocene planktonic foraminifer biostratigraphy of ODP Leg 135 sites

Diverse and well-preserved planktonic foraminifers were recovered from six sites (834-839) drilled in the Lau Basin. Planktonic faunas from the Tongan Platform sites varied from those of the Lau Basin sites by being less well preserved (Site 840) to being very poorly preserved and very sparse (Site 841); at Site 841 most samples were barren. All sites penetrated a volcaniclastic sequence in which thick ash beds were encountered; foraminifer populations within the ash beds were often very small, making it difficult to obtain biostratigraphic data. No hiatuses were encountered in the upper Miocene to Pleistocene sections of the Lau Basin, but a possible break occurs at Site 840 on the Tongan Platform. Site 834 penetrated through a Quaternary-Pliocene sequence overlying basaltic basement, and topmost Miocene (Zone N17B) sediments interbedded within the volcanic sequence. Site 835 penetrated into the lower Pliocene (Zones N19 to N19-20). Site 836 penetrated the shortest section, with Zone N22 {Globorotalia (Truncorotalia) crassaformis hessi Subzone) directly overlying basalts. Site 837 penetrated into the basal part of Zone N22 (Globigerinoides quadrilobatus fistulosus Subzone) overlying basalt. Site 838 failed to encounter basalts, with the oldest sediment being from Zone N22 (Globigerinoides quadrilobatus fistulosus Subzone). Site 839, within the same basin as Site 838, located Zone N22 (Globigerinoides quadrilobatus fistulosus Subzone) sediments directly overlying igneous basement. Site 840 penetrated into the upper Miocene Zone N17A without encountering any major unconformity. Site 841, studied mainly from core-catcher samples, penetrated a Quaternary to questionable upper Miocene sequence that was in fault contact with middle Miocene (Zones N8 to N9) sediments. For the Lau Basin sites, reworking was encountered only in Sites 834 and 835. Site 834 was drilled adjacent to the Lau Ridge, on which are developed numerous reef al and shallow-water environments, where erosional conditions could have been expected during sea-level lowstands. Site 835 was drilled in a narrow basin that has been remote from these erosional influences; slumping and erosion of material from the adjacent basin slopes appears to have been the source of the reworking. For the Tongan Platform sites, reworking was observed only in the lower part of the upper Miocene section at Site 841, where late Eocene larger foraminifers are present in conglomerates and grits. The presence of Globorotalia (Globorotalia) multicamerata and small specimens of Sphaeroidinellopsis spp. in the Pleistocene of Site 840 may indicate reworking, but this is not clear. Unit I, which marks a reduction in volcanic activity in the Lau Basin, ranges in age from the lower part of Zone N22 (Globigerinoides quadrilobatus fistulosus Subzone) at Sites 834 and 835, to within Zone N22 (Globorotalia crassaformis hessi Subzone) at Sites 836 to 838, and within the upper part of Zone N22 (Bolliella praeadamsi Subzone) at Site 839. Units II and III are generally represented by thick to very thick ash beds, which generally contain low-diversity and often poorly preserved assemblages. Igneous sources seem to have remained important contributors of sediment up to the present day.

Geochemistry of ODP Leg 135 basalts

Cr-spinels in cores drilled during Ocean Drilling Program Leg 135 exhibit wide variations in composition and morphology that reflect complex petrogenetic histories. These Cr-spinels are found within basaltic lava flows that erupted in north-trending sub-basins within the Lau Basin backarc. Cr-spinels from Sites 834 and 836 occur as euhedral groundmass grains and inclusions in plagioclase, and range up to 300 ?m in size. These Cr-spinels are similar in composition, morphology, and mode of occurrence to Cr-spinels found within depleted, N-type mid-ocean-ridge basalts (N-MORB), reflecting similar crystallization conditions and host lava composition to N-MORB. Their compositional range is relatively narrow, with Cr/(Cr + Al + Fe3+) (Cr#) and Mg/(Mg + Fe2+) (Mg#) varying from 0.38 to 0.48 and 0.56 to 0.72, respectively; like Cr-spinels from N-MORB, they contain low amounts of TiO2 (0.37%-1.05%) and Fe3+/(Cr + Al + Fe3+) (Fe3+#; <0.11). In contrast, Cr-spinels from Site 839 have much higher Cr# at a given Mg#, with Cr# varying from 0.52 to 0.76 and Mg# varying from 0.27 to 0.75. These Cr-spinels are similar in composition to those from primitive, boninitic or low-Al2O3 arc basalts, sharing their low TiO2 and Fe3+# (typically below 0.35% and 0.1, respectively for spinel grain interiors). Site 839 Cr-spinels occur as small (to 50 µm) euhedra within strongly zoned olivine or as unusually large (to 3 mm), euhedral to subhedral megacrysts. These megacrysts are strongly zoned in Mg#, but they display little zoning in Cr#, providing evidence of strong compositional disequilibria with the host melt. The magnesian cores of the megacrysts crystallized from primitive, near-primary melts derived from harzburgitic or highly depleted lherzolitic sources, and they provide evidence that the Site 839 spinel-bearing lavas were derived by the mixing of melt with a Mg# of 0.75-0.80 and evolved, Cr-spinel barren melt with a Mg# < 0.6 shortly before eruption.

(Table 1) Platinum group element abundanves in ODP Leg 135 samples

Ocean Drilling Program Leg 135 backarc basin lavas are characterized by anomalously high Au contents (1.0-11.4 ppb) and strongly fractionated relative platinum group element (PGE) abundances (Pd/Ir ratio, approximately 100). The Rh and Ir contents are very low, ranging from below detection (approximately 0.02 ppb) to 0.08 ppb. The Pd and Pt contents range from <0.3 to 4 ppb. Rh, Pd, and Pt values are consistently and significantly higher in Site 836 and 839 samples relative to those from Sites 834 and 835. Major, trace, and rare earth element (REE) data suggest Sites 836 and 839 have a more pronounced arc signature than Sites 834 and 835. No correlation exists between noble metal abundance and indices of alteration or fractionation (e.g., loss on ignition (LOI), Mg#, and Cr or Ni contents), suggesting that measured values and ratios are primary and reflect characteristics of the mantle source. The evaluation of Leg 135 noble metal data with respect to potential mantle-source components is hindered by the lack of data on magmas derived from such sources. However, analyses of the limited available data for the different magma types suggest that the characteristic enrichment of Leg 135 lavas in Au, relative to Pd and Cu, cannot be derived solely from simple MORB-type or ocean-island-type mantle, or mantle depleted by a previous melt extraction event. The Au-enriched signature of the Lau basin lavas could, however, be produced through the addition of a sedimentary component from the downgoing slab. Separation of Au from the PGE occurs within oceanic hydrothermal systems and gold values of the resultant precipitates are 2-3 orders of magnitude higher than other oceanic crustal components. Even small additions of this component from the downgoing oceanic crust to a supra-subduction zone mantle melt could account for the high mean Au/Pd ratios of the Leg 135 samples (Sites 834 and 835, Au/Pd = 5.04; Sites 836 and 839, Au/Pd = 2.26).

In vitro and in vivo examination of the cell surface glycoprotein CDCP1

A number of reports have demonstrated the importance of the CUB domaincontaining protein 1 (CDCP1) in facilitating cancer progression in animal models and the potential of this protein as a prognostic marker in several malignancies. CDCP1 facilitates metastasis formation in animal models by negatively regulating anoikis, a type of apoptosis triggered by the loss of attachment signalling from cell-cell contacts or cell-extra cellular matrix (ECM) contacts. Due to the important role CDCP1 plays in cancer progression in model systems, it is considered a potential drug target to prevent the metastatic spread of cancers. CDCP1 is a highly glycosylated 836 amino acid cell surface protein. It has structural features potentially facilitating protein-protein interactions including 14 N-glycosylation sites, three CUB-like domains, 20 cysteine residues likely to be involved in disulfide bond formation and five intracellular tyrosine residues. CDCP1 interacts with a variety of proteins including Src family kinases (SFKs) and protein kinase C ä (PKCä). Efforts to understand the mechanisms regulating these interactions have largely focussed on three CDCP1 tyrosine residues Y734, Y743 and Y762. CDCP1-Y734 is the site where SFKs phosphorylate and bind to CDCP1 and mediate subsequent phosphorylation of CDCP1-Y743 and -Y762 which leads to binding of PKCä at CDCP1-Y762. The resulting trimeric protein complex of SFK•CDCP1•PKCä has been proposed to mediate an anti-apoptotic cell phenotype in vitro, and to promote metastasis in vivo. The effect of mutation of the three tyrosines on interactions of CDCP1 with SFKs and PKCä and the consequences on cell phenotype in vitro and in vivo have not been examined. CDCP1 has a predicted molecular weight of ~90 kDa but is usually detected as a protein which migrates at ~135 kDa by Western blot analysis due to its high degree of glycosylation. A low molecular weight form of CDCP1 (LMWCDCP1) of ~70 kDa has been found in a variety of cancer cell lines. The mechanisms leading to the generation of LMW-CDCP1 in vivo are not well understood but an involvement of proteases in this process has been proposed. Serine proteases including plasmin and trypsin are able to proteolytically process CDCP1. In addition, the recombinant protease domain of the serine protease matriptase is also able to cleave the recombinant extracellular portion of CDCP1. Whether matriptase is able to proteolytically process CDCP1 on the cell surface has not been examined. Importantly, proteolytic processing of CDCP1 by trypsin leads to phosphorylation of its cell surface-retained portion which suggests that this event leads to initiation of an intracellular signalling cascade. This project aimed to further examine the biology of CDCP1 with a main of focus on exploring the roles played by CDCP1 tyrosine residues. To achieve this HeLa cells stably expressing CDCP1 or the CDCP1 tyrosine mutants Y734F, Y743F and Y762F were generated. These cell lines were used to examine: • The roles of the tyrosine residues Y734, Y743 and Y762 in mediating interactions of CDCP1 with binding proteins and to examine the effect of the stable expression on HeLa cell morphology. • The ability of the serine protease matriptase to proteolytically process cell surface CDCP1 and to examine the consequences of this event on HeLa cell phenotype and cell signalling in vitro. • The importance of these residues in processes associated with cancer progression in vitro including adhesion, proliferation and migration. • The role of these residues on metastatic phenotype in vivo and the ability of a function-blocking anti-CDCP1 antibody to inhibit metastasis in the chicken embryo chorioallantoic membrane (CAM) assay. Interestingly, biochemical experiments carried out in this study revealed that mutation of certain CDCP1 tyrosine residues impacts on interactions of this protein with binding proteins. For example, binding of SFKs as well as PKCä to CDCP1 was markedly decreased in HeLa-CDCP1-Y734F cells, and binding of PKCä was also reduced in HeLa-CDCP1-Y762F cells. In contrast, HeLa-CDCP1-Y743F cells did not display altered interactions with CDCP1 binding proteins. Importantly, observed differences in interactions of CDCP1 with binding partners impacted on basal phosphorylation of CDCP1. It was found that HeLa-CDCP1, HeLa-CDCP1-Y743F and -Y762F displayed strong basal levels of CDCP1 phosphorylation. In contrast, HeLa-CDCP1-Y734F cells did not display CDCP1 phosphorylation but exhibited constitutive phosphorylation of focal adhesion kinase (FAK) at tyrosine 861. Significantly, subsequent investigations to examine this observation suggested that CDCP1-Y734 and FAK-Y861 are competitive substrates for SFK-mediated phosphorylation. It appeared that SFK-mediated phosphorylation of CDCP1- Y734 and FAK-Y861 is an equilibrium which shifts depending on the level of CDCP1 expression in HeLa cells. This suggests that the level of CDCP1 expression may act as a regulatory mechanism allowing cells to switch from a FAK-Y861 mediated pathway to a CDCP1-Y734 mediated pathway. This is the first time that a link between SFKs, CDCP1 and FAK has been demonstrated. One of the most interesting observations from this work was that CDCP1 altered HeLa cell morphology causing an elongated and fibroblastic-like appearance. Importantly, this morphological change depended on CDCP1- Y734. In addition, it was observed that this change in cell morphology was accompanied by increased phosphorylation of SFK-Y416. This suggests that interactions of SFKs with CDCP1-Y734 increases SFK activity since SFKY416 is critical in regulating kinase activity of these proteins. The essential role of SFKs in mediating CDCP1-induced HeLa cell morphological changes was demonstrated using the SFK-selective inhibitor SU6656. This inhibitor caused reversion of HeLa-CDCP1 cell morphology to an epithelial appearance characteristic of HeLa-vector cells. Significantly, in vitro studies revealed that certain CDCP1-mediated cell phenotypes are mediated by cellular pathways dependent on CDCP1 tyrosine residues whereas others are independent of these sites. For example, CDCP1 expression caused a marked increase in HeLa cell motility that was independent of CDCP1 tyrosine residues. In contrast, CDCP1- induced decrease in HeLa cell proliferation was most prominent in HeLa- CDCP1-Y762F cells, potentially indicating a role for this site in regulating proliferation in HeLa cells. Another cellular event which was identified to require phosphorylation of a particular CDCP1 tyrosine residue is adhesion to fibronectin. It was observed that the CDCP1-mediated strong decrease in adhesion to fibronectin is mostly restored in HeLa-CDCP1-Y743F cells. This suggests a possible role for CDCP1-Y743 in causing a CDCP1-mediated decrease in adhesion. Data from in vivo experiments indicated that HeLa-CDCP1-Y734F cells are more metastic than HeLa-CDCP1 cells in vivo. This indicates that interaction of CDCP1 with SFKs and PKCä may not be required for CDCP1-mediated metastasis formation of HeLa cells in vivo. The metastatic phenotype of these cells may be caused by signalling involving FAK since HeLa-CDCP1- Y734F cells are the only CDCP1 expressing cells displaying constitutive phosphorylation of FAK-Y861. HeLa-CDCP1-Y762F cells displayed a very low metastatic ability which suggests that this CDCP1 tyrosine residue is important in mediating a pro-metastatic phenotype in HeLa cells. More detailed exploration of cellular events occurring downstream of CDCP1-Y734 and -Y762 may provide important insights into the mechanisms altering the metastatic ability of CDCP1 expressing HeLa cells. Complementing the in vivo studies, anti-CDCP1 antibodies were employed to assess whether these antibodies are able to inhibit metastasis of CDCP1 and CDCP1 tyrosine mutants expressing HeLa cells. It was found that HeLa- CDCP1-Y734F cells were the only cell line which was markedly reduced in the ability to metastasise. In contrast, the ability of HeLa-CDCP1, HeLa- CDCP1-Y743F and -Y762F cells to metastasise in vivo was not inhibited. These data suggest a possible role of interactions of CDCP1 with SFKs, occurring at CDCP1-Y734, in preventing an anti-metastatic effect of anti- CDCP1 antibodies in vivo. The proposal that SFKs may play a role in regulating anti-metastatic effects of anti-CDCP1 antibodies was supported by another experiment where differences between HeLa-CDCP1 cells and CDCP1 expressing HeLa cells (HeLa-CDCP1-S) from collaborators at the Scripps Research Institute were examined. It was found that HeLa-CDCP1-S cells express different SFKs than CDCP1 expressing HeLa cells generated for this study. This is important since HeLa-CDCP1-S cells can be inhibited in their metastatic ability using anti-CDCP1 antibodies in vivo. Importantly, these data suggest that further examinations of the roles of SFKs in facilitating anti-metastatic effects of anti-CDCP1 antibodies may give insights into how CDCP1 can be blocked to prevent metastasis in vivo. This project also explored the ability of the serine protease matriptase to proteolytically process cell surface localised CDCP1 because it is unknown whether matriptase can cleave cell surface CDCP1 as it has been reported for other proteases such as trypsin and plasmin. Furthermore, the consequences of matriptase-mediated proteolysis on cell phenotype in vitro and cell signalling were examined since recent reports suggested that proteolysis of CDCP1 leads to its phosphorylation and may initiate cell signalling and consequently alter cell phenotype. It was found that matriptase is able to proteolytically process cell surface CDCP1 at low nanomolar concentrations which suggests that cleavage of CDCP1 by matriptase may facilitate the generation of LWM-CDCP1 in vivo. To examine whether matriptase-mediated proteolysis induced cell signalling anti-phospho Erk 1/2 Western blot analysis was performed as this pathway has previously been examined to study signalling in response to proteolytic processing of cell surface proteins. It was found that matriptase-mediated proteolysis in CDCP1 expressing HeLa cells initiated intracellular signalling via Erk 1/2. Interestingly, this increase in phosphorylation of Erk 1/2 was also observed in HeLa-vector cells. This suggested that initiation of cell signalling via Erk 1/2 phosphorylation as a result of matriptase-mediated proteolysis occurs by pathways independent of CDCP1. Subsequent investigations measuring the flux of free calcium ions and by using a protease-activated receptor 2 (PAR2) agonist peptide confirmed this hypothesis. These data suggested that matriptase-mediated proteolysis results in cell signalling via a pathway induced by the activation of PAR2 rather than by CDCP1. This indicates that induction of cell signalling in HeLa cells as a consequence of matriptase-mediated proteolysis occurs via signalling pathways which do not involve phosphorylation of Erk 1/2. Consequently, it appears that future attempts should focus on the examination of cellular pathways other than Erk 1/2 to elucidate cell signalling initiated by matriptase-mediated proteolytic processing of CDCP1. The data presented in this thesis has explored in vitro and in vivo aspects of the biology of CDCP1. The observations summarised above will permit the design of future studies to more precisely determine the role of CDCP1 and its binding partners in processes relevant to cancer progression. This may contribute to further defining CDCP1 as a target for cancer treatment.

Review: "Context : A framework for its Influence on Evaluation Practice (New Directions for Evaluation No. 135)" edited by Rog, D. J.; Fitzpatrick, J. L. and Conner, R. F.

This book offers a framework for the influence of context on evaluation practice and is applied to three case studies: environmental context; indigenous context and political context; and finishes with a process for implementation.