The Role Of IL-8-Mediated Src Family Kinase Activation In Tumor-Tumor And Tumor-Stromal Interactions In Metastasis Of Prostate Cancer

Men with localized prostate cancer (PCa) have a 100% five-year survival rate, but this rate drops to 33% for men with metastatic disease. A better understanding of the metastatic process is needed to develop better therapies for PCa. Aberrant activation of protein tyrosine kinases, including Src Family Kinases (SFKs) contribute to metastasis through numerous functions, one of which leads to increased expression of cytokines, such as IL-8. However, the relationship between Src activity and IL-8 regulation is not completely understood. In cell line models, I determined that IL-8 activates Src and in turn Src activates IL-8 demonstrating a feed forward loop contributing to the migration and invasion of PCa cells. However, IL-8 is also produced by tumor-associated stromal cells. In bone marrow derived stromal cells (HS5), I demonstrated a feed forward loop occurs as was observed in tumor cells. HS5 conditioned media increased Src activity in PCa cells. By silencing IL-8 in HS5 cells, Src activity was decreased to control levels in PCa cells as was migration and invasion. Thus, stromal cells producing IL-8 contribute to metastatic properties of PCa by a paracrine mechanism. To examine the effect of stromal cells on tumor growth and metastatic potential of PCa in vivo, I mixed HS5 and PCa cells and co-injected them intraprostatically. I determined that tumor growth and metastases were increased. By silencing IL-8 in HS5 cells and co-injecting them with PCa cells intraprostatically, tumor growth and metastases were still increased relative to injection of PCa cells alone, but decreased relative to co-injections with PCa cells and HS5 cells. These studies demonstrated: (1) a feed forward loop in both tumor and stromal cells, whereby IL-8 activates Src, derepressing IL-8 expression in PCa cells in vitro; (2) stromal produced IL-8 activates Src and contributes to the migration and invasion of PCa cells in vitro; and (3) stromal produced IL-8 is responsible, in part, for increases in PCa tumor growth and metastatic potential. Together, these studies demonstrated that IL-8-mediated Src activity increases the metastatic potential of PCa and therapeutic agents interfering with the IL-8/SFK signaling axis may be useful for prevention and treatment of metastases.

Structure and biosynthesis of a pyruvate -dependent enzyme---phosphatidylserine decarboxylase from {\it Escherichia coli\/}

Phosphatidylserine decarboxylase of E. coli, a cytoplasmic membrane protein, catalyzes the formation of phosphatidylethanolamine, the principal phospholipid of the organism. The activity of the enzyme is dependent on a covalently bound pyruvate (Satre and Kennedy (1978) J. Biol. Chem. 253, 479-483). This study shows that the enzyme consists of two nonidentical subunits, $\alpha$ (Mr = 7,332) and $\beta$ (Mr = 28,579), with the pyruvate prosthetic group in amide linkage to the amino-terminus of the $\alpha$ subunit. Partial protein sequence and DNA sequence analysis reveal that the two subunits are derived from a proenzyme ($\pi$ subunit, Mr = 35,893) through a post-translational event. During the conversion of the proenzyme to the $\alpha$ and $\beta$ subunits, the peptide bond between Gly253-Ser254 is cleaved, and Ser254 is converted to the pyruvate prosthetic group at the amino-terminus of the $\alpha$ subunit (Li and Dowhan (1988) J. Biol. Chem. 263, 11516-11522).^ The proenzyme cannot be detected in cells carrying either single or multiple copies of the gene (psd), but can be observed in a T7 RNA polymerase/promoter and transcription-translation system. The cleavage of the wild-type proenzyme occurs rapidly with a half-time on the order of 2 min. Changing of the Ser254 to cysteine (S254C) or threonine (S254T) slows the cleavage rate dramatically and results in mutants with a half-time for processing of around 2-4 h. Change of the Ser254 to alanine (S254A) blocks the cleavage of the proenzyme. The reduced processing rate with the mutations of the proenzyme is consistent with less of the functional enzyme being made. Mutants S254C and S254T produce $\sim$15% and $\sim$1%, respectively, of the activity of the wild-type allele, but can still complement a temperature-sensitive mutant of the psd locus. Neither detectable activity nor complementation is observed by mutant S254A. These results are consistent with the hydroxyl-group of the Ser254 playing a critical role in the cleavage of the peptide bond Gly253-Ser254 of the pro-phosphatidylserine decarboxylase, and support the mechanism proposed by Snell and co-workers (Recsei and Snell (1984) Annu. Rev. Biochem. 53, 357-387) for the formation of the prosthetic group of pyruvate-dependent decarboxylases. ^

Functional studies of *Ras and Bcl-2 oncoproteins in keratinocyte homeostasis and multistep skin carcinogenesis

To assess the effect of deregulated Ha-ras and bcl-2, individually and in combination on epidermal keratinocyte homeostasis and during multistep skin carcinogenesis, we generated skin-specific transgenic mice and keratinocyte transfectants constitutively expressing oncogenic Ha-ras and bcl-2 proteins. The deregulated Ha-ras and bcl-2 expression contributing to homeostatic imbalances in the skin had an additive effect on the probability of tumor development. They were also cooperative in incidence, growth, and latency of tumor formation, and they exhibited synergistic cooperation in malignant transformation of benign papillomas. To explain the homeostatic imbalances by Ha-ras and bcl-2 overexpression in the skin, we investigated the three major cellular processes of proliferation, cell death, and differentiation. Epidermal expression of Bcl-2 retarded keratinocyte proliferation in the epidermis of neonatal mice compared with results for control littermates. Constitutive expression of Ha-ras increased keratinocyte proliferation, and co-expression of bcl-2 modestly suppressed the ras-mediated abnormal proliferation of neonatal keratinocytes. Bcl-2 proteins in keratinocytes protected UV-treated cells from apoptotic cell death regardless of oncogenic ras expression in both non-neoplastic neonatal epidermis and human keratinocyte cell lines. The spontaneous apoptotic index (AI) was also lower in papillomas constitutively expressing bcl-2 compared with the ones that developed in control mice. Ras-overexpressing epidermis, including that in ras/bcl-2 double transgenic mice, had abnormal differentiation patterns compared with controls. The oncogenic ras protein had alterations in both epidermal distribution and the extent of cytokeratin 14 and involucrin expression. Abnormal expression of the hyperproliferation marker cytokeratin 6 and modest down regulation of cytokeratin 1 were also detected. Late appearance of filaggrin was another abnormal phenotype of the ras-expressing epidermis. Overexpression of bcl-2 had no effect on epidermal differentiation. Together, these findings suggest that constitutive expression of oncogenic Ha-ras and bcl-2 are important determinants of epidermal proliferation, viability and differentiation. In summary, our results demonstrated that the disruption of epidermal homeostasis by overexpressed ras and bcl-2 predisposes to hyperplastic growth of the epidermis and to papilloma development and that these proteins with distinct mechanisms for oncogenesis are functionally synergistic for malignant transformation of chemically induced skin carcinogenesis. ^

(Table 1) Contents of tungsten and other metals in Fe-Mn nodules from the Pacific Ocean

Tungsten contents in iron-manganese nodules and crusts from different parts of the World Ocean, as well as its relationships with a number of chemical elements are under consideration. A trend to correlation of tungsten with Fe, Ti, W, Pb, and Co is noticed. Comparison of tungsten contents in the nodules and host sediments indicates its low geochemical mobility.

Composition of Fe-Mn micronodules, nodules and host bottom sediments from the Northeast Pacific Basin

Authigenic ferromanganese manifestations in bottom sediments from two horizons (0-10 and 240-250 cm) located in the low/high bioproductive transitional zone of the Pacific Ocean were studied. In addition two compositionally different types of micronodules, crusts and ferromanganese nodules were detected in the surface horizon (0-1 cm). Three size fractions (50-100, 100-250, and 250-500 µm) of manganese micronodules were investigated. In terms of surface morphology, color, and shape, the micronodules are divided into dull round (MN1) and angular lustrous (MN2) varieties with different mineral and chemical compositions. MN1 are enriched in Mn and depleted in Fe as compared with MN2. Mn/Fe ratio in MN1 varies from 13 to 14. Asbolane-buserite and birnessite are the major manganese minerals in them. MN2 is mainly composed of vernadite with Mn/Fe ratio from 4.3 to 4.8. Relative to MN1, fraction 50-100 µm of MN2 is enriched in Fe (2.6 times), W (1.8), Mo (3.2), Th (2.3), Ce (5.8), and REE (from 1.2 to 1.8). Relative to counterparts from MN1, separate fractions of MN2 are characterized by greater compositional difference. For example, increase in size of micronodules leads to decrease in contents of Fe (by 10 rel. %), Ce (2 times), W (2.1 times), Mo (2.2 times), and Co (1.5 times). At the same time one can see increase in contents of other elements: Th and Cu (2.1 times), Ni (1.9 times), and REE (from 1.2 to 1.6 times). Differences in chemical and mineral compositions of MN1 and MN2 fractions can be related to alternation of oxidative and suboxidative conditions in the sediments owing to input of labile organic matter, which acts as the major reducer, and allochthonous genesis of MN2.

Environmental variables and multivariate seabed classification maps via k means clustering of Potter Cove, Antarctica, link to input and result files in GeoTIFF format

This study subdivides the Potter Cove, King George Island, Antarctica, into seafloor regions using multivariate statistical methods. These regions are categories used for comparing, contrasting and quantifying biogeochemical processes and biodiversity between ocean regions geographically but also regions under development within the scope of global change. The division obtained is characterized by the dominating components and interpreted in terms of ruling environmental conditions. The analysis includes in total 42 different environmental variables, interpolated based on samples taken during Australian summer seasons 2010/2011 and 2011/2012. The statistical errors of several interpolation methods (e.g. IDW, Indicator, Ordinary and Co-Kriging) with changing settings have been compared and the most reasonable method has been applied. The multivariate mathematical procedures used are regionalized classification via k means cluster analysis, canonical-correlation analysis and multidimensional scaling. Canonical-correlation analysis identifies the influencing factors in the different parts of the cove. Several methods for the identification of the optimum number of clusters have been tested and 4, 7, 10 as well as 12 were identified as reasonable numbers for clustering the Potter Cove. Especially the results of 10 and 12 clusters identify marine-influenced regions which can be clearly separated from those determined by the geological catchment area and the ones dominated by river discharge.

Composition of dissolved, particulate and sinking material in hydrothermal plumes above the TAG and Broken Spur hydrothermal fields

The paper reports data on distribution of dissolved (Mn, Zn, Cu, Pb, and Cd) and particulate (Fe, Mn, Zn, Cu, Pb, Ni, and Co) species of metals in hydrothermal plumes above the active TAG and Broken Spur hydrothermal fields (26° N and 29° N in the MAR rift valley, respectively). Sediment trap data on fluxes of hydrothermal sedimentary material in the areas indicate that (i) the predominant Zn source for metalliferous sediments at the TAG field is material precipitating from the neutrally buoyant plume, and (ii) the predominant source of Fe and Co is re-deposited ore material coming from the area of extensive settling of sulfides.

Major and minor element concentrations in the livers of polar bears (Ursus maritimus) from Greenland, Canada and Alaska

To assess geographic distributions of elements in the Arctic we compared essential and non-essential elements in the livers of polar bears (Ursus maritimus) collected from five regions within Canada in 2002, in Alaska between 1994 and 1999 and from the northwest and east coasts of Greenland between 1988 and 2000. As, Hg, Pb and Se varied with age, and Co and Zn with gender, which limited spatial comparisons across all populations to Cd, which was highest in Greenland bears. Collectively, geographic relationships appeared similar to past studies with little change in concentration over time in Canada and Greenland for most elements; Hg and Se were higher in some Canadian populations in 2002 as compared to 1982 and 1984. Concentrations of most elements in the polar bears did not exceed toxicity thresholds, although Cd and Hg exceeded levels correlated with the formation of hepatic lesions in laboratory animals.

Description and geochemical composition of manganese nodules retrieved from Lake Ontario, Canada

A large deposit of ferromanganese oxide coated sands and scattered manganese nodules occurs in the northern portion of Lake Ontario. The Mn and Fe contents of the concretions are similar to those in concretions from other environments, while their Ni, Cu, and Co contents are lower than in deep-sea nodules, but higher than in most previously described lacustrine concretions. Pb and Zn are high in the coatings and exceed the concentrations found in many previously analyzed Mn deposits. Within the deposit, Mn, Ni, Co, and Zn contents are correlated, and they vary inversely with Fe. Mn, Fe, Ni, Cu, and Pb are present in the interstitial waters of the sediments underlying the deposit in higher concentrations than in the overlying lake waters, thus providing a potential source of metals for concretion formation.The origin and compositional variations in the deposit possibly can be explained in terms of the fractionation and precipitation of Fe and Mn as a result of redox variations in the lake sediments. Eh increases from south to north across the deposit in such a way that iron may be selectively oxidized and precipitated in the south and manganese, in the north. The upward diffusion of Mn, Fe, and associated elements from the underlying sediments probably provides the principal source of the metals in the south of the deposit, while metal-enriched bottom waters are probably the principal source in the north.

Trace and rare elements and factors at DSDP Hole 71-511

The distribution of paragenetic assemblages of trace and rare elements, as revealed by factor analysis (R-mode, Q-mode), the ratios of elements to Zr and the interpretation of these data in the context of the known mineralogy, lithology, and geology of the region, provide the bases for the outline of the geochemical history of sedimentation in the study area that forms the subject of this chapter. Two stages may be discerned. 1. Late-Middle Jurassic-Early Cretaceous (160-106? Ma). The sediments that accumulated in relatively shallow water (shelf) were predominantly clay, with dispersed sapropelic organic matter, plant fragments, pyrite, admixtures of acid-medium volcanic glass, and epigenetic crystals of gypsum. The bottom water layers of the basin are notably stagnant. The sediments are characterized by higher amounts of V, Zn, Cu, Cr, Rb, and Be associated with organic matter. Lower Cretaceous sediments, separated from those of the Upper Jurassic by a hiatus, accumulated in a deepened and enlarging basin. These Lower Cretaceous deposits are chemically similar to those of the Upper Jurassic, but contain diagenetic concentrations of Zn, Ni, and La. 2. Early-middle Albian (Unit 5)-middle Maestrichtian (1067-66.6Ma). The prevailing regime was that of an open ocean basin that tended to expand and deepen. During the second half of the early-middle Albian, the biogenic components Ba, Sr, and CaCO3 accumulated. By the end of this interval, Ti/Zr values had increased. In conjunction data on mineral composition, they testify to an outburst of basaltoid volcanism related to tectonic activity before an erosional hiatus (late Albian-Cenomanian). At the end of the Cenomanian-Turonian, residual deposits of predominantly clay sediments with relatively high amounts of Ti and Zr and associated rare alkalis (Li, Rb) accumulated. Clay sediments deposited during the Coniacian-Santonian were characterized by higher concentrations of Ti, Zr, Li, and Rb, by diagenetic carbonate phases of Ni, Zn, and La, and by sulphides and Fe-oxides with an admixture of Ni and Co. The latter half of the interval saw the deposition of fine basaltoid volcanoclastic material, diagenetically altered by zeolitization and carbonatization and enriched with Se, Pb, Ti, Sr, Ba, Y, and Yb. Sediments with a similar chemistry accumulated in the Campanian-middle Maestrichtian. Strong current activity preceding a global hiatus at the Mesozoic/Cenozoic boundary is reflected in both lower sedimentation rates and the presence of higher residual concentrations of Ti, Zr, Ba, Sr, and other elements studied in this chapter.

Investigations on fossil bivalves and wood from Seymour Island, Antarctica

Quasi-periodic variation in sea-surface temperature, precipitation, and sea-level pressure in the equatorial Pacific known as the El Niño - Southern Oscillation (ENSO) is an important mode of interannual variability in global climate. A collapse of the tropical Pacific onto a state resembling a so-called 'permanent El Niño', with a preferentially warmed eastern equatorial Pacific, flatter thermocline, and reduced interannual variability, in a warmer world is predicted by prevailing ENSO theory. If correct, future warming will be accompanied by a shift toward persistent conditions resembling El Niño years today, with major implications for global hydrological cycles and consequent impacts on socioeconomic and ecological systems. However, much uncertainty remains about how interannual variability will be affected. Here, we present multi-annual records of climate derived from growth increment widths in fossil bivalves and co-occurring driftwood from the Antarctic peninsula that demonstrate significant variability in the quasi-biennial and 3-6 year bands consistent with ENSO, despite early Eocene (~50 Mya) greenhouse conditions with global average temperature -10 degrees higher than today. A coupled climate model suggests an ENSO signal and teleconnections to this region during the Eocene, much like today. The presence of ENSO variation during this markedly warmer interval argues for the persistence of robust interannual variability in our future greenhouse world.

Average fluorescence and dissolved iron and Fe-binding ligand characteristics during POLARSTERN expedition ANT-XXIV/3 in 2008

Organic complexation of dissolved iron (dFe) was investigated in the Atlantic sector of the Southern Ocean in order to understand the distribution of Fe over the whole water column. The total concentration of dissolved organic ligands ([Lt]) measured by voltammetry ranged between 0.54 and 1.84 nEq of M Fe whereas the conditional binding strength (K') ranged between 10**21.4 and 10**22.8. For the first time, trends in Fe-organic complexation were observed in an ocean basin by examining the ratio ([Lt]/[dFe]), defined as the organic ligand concentration divided by the dissolved Fe concentration. The [Lt]/[dFe] ratio indicates the saturation state of the natural ligands with Fe; a ratio near 1 means saturation of the ligands leading to precipitation of Fe. Reversely, high ratios mean Fe depletion and show a high potential for Fe solubilisation. In surface waters where phytoplankton is present low dissolved Fe and high variable ligand concentrations were found. Here the [Lt]/[dFe] ratio was on average 4.4. It was especially high (5.6-26.7) in the HNLC (High Nutrient, Low Chlorophyll) regions, where Fe was depleted. The [Lt]/[dFe] ratio decreased with depth due to increasing dissolved Fe concentrations and became constant below 450 m, indicating a steady state between ligand and Fe. Relatively low [Lt]/[dFe] ratios (between 1.1 and 2.7) existed in deep water north of the Southern Boundary, facilitating Fe precipitation. The [Lt]/[dFe] ratio increased southwards from the Southern Boundary on the Zero Meridian and from east to west in the Weddell Gyre due to changes both in ligand characteristics and in dissolved iron concentration. High [Lt]/[dFe] ratio expresses Fe depletion versus ligand production in the surface. The decrease with depth reflects the increase of [dFe] which favours scavenging and (co-) precipitation, whereas a horizontal increase in the deep waters results from an increasing distance from Fe sources. This increase in the [Lt]/[dFe] ratio at depth shows the very resistant nature of the dissolved organic ligands.

Benthic foraminifera and sea level analyses from IODP Hole 310-M0005D

The course of sea-level fluctuations during Termination II (TII; the penultimate deglaciation), which is critical for understanding ice-sheet dynamics and suborbital climate variability, has yet to be established. This is partly because most shallow-water sequences encompassing TII were eroded during sea-level lowstands of the last glacial period or were deposited below the present sea level. Here we report a new sequence recording sea-level changes during TII in the Pleistocene sequence at Hole M0005D (water depth: 59.63 m below sea level [mbsl]) off Tahiti, French Polynesia, which was drilled during Integrated Ocean Drilling Program Expedition 310. Lithofacies variations and stratigraphic changes in the taxonomic composition, preservation states, and intraspecific test morphology of large benthic foraminifers indicate a deepening-upward sequence in the interval from Core 310-M0005D-26R (core depth: 134 mbsl) through -16R (core depth: 106 mbsl). Reconstruction of relative sea levels, based on paleodepth estimations using large benthic foraminifers, indicated a rise in sea level of about 90 m during this interval, suggesting its correlation with one of the terminations. Assuming that this rise in sea level corresponds to that during TII, after correcting for subsidence since the time of deposition, a highstand sea-level position would be 2 ± 15 m above present sea level (masl), which is generally consistent with highstand sea-level positions in MIS 5e (4 ± 2 masl). If this rise in sea level corresponds to that during older terminations, the subsidence-corrected highstand sea-level positions (30 ± 15 masl for Termination III and 54 ± 15 masl for Termination IV) are not consistent with reported ranges of interglacial sea-level highstands (-18 to 15 masl). Therefore, the studied interval likely records the rise in sea level and associated environmental changes during TII. In particular, the intervening cored materials between the two episodes of sea-level rise found in the studied interval might record the sea-level reversal event during TII. This conclusion is consistent with U/Th ages of around 133 ka, which were obtained from slightly diagenetically altered (i.e., < 1% calcite) in situ corals in the studied interval (Core 310-M0005D-20R [core depth: 118 mbsl]). This study also suggests that our inverse approach to correlate a stratigraphic interval with an approximate time frame could be useful as an independent check on the accuracy of uranium-series dating, which has been applied extensively to fossil corals in late Quaternary sea-level studies.

Magnetobiostratigraphy and stable isotope record of Paleocene sediments from ODP Hole 122-761B in the subtropical Indian Ocean

Surface and deep water circulation patterns in the eastern Indian Ocean during the Paleocene Epoch are inferred based on an integrated magnetobiostratigraphic and stable isotope investigation of Ocean Drilling Program Hole 761B, drilled on the Wombat Plateau. A combination of magnetostratigraphy, biostratigraphy and isotope stratigraphy demonstrates that numerous deep sea sites that have been considered to show continuous, or nearly continuous sedimentation through the Paleocene are punctuated by a series of hiatuses, some of which exceeding a duration of 1 Myr. Therefore, our study is based on a detailed temporal interpretation of the stratigraphic successions we used for paleoceanographic reconstructions. We compare detailed planktonic and benthic foraminiferal carbon and oxygen isotope records from Hole 761B with several temporally correlative records published from different oceanic provinces in order to distinguish between local and global patterns within the eastern Indian Ocean. Although Site 761 was situated at low latitudes during the Paleocene, its surface waters were predominantly influenced by circulation originating from the Southern Ocean as indicated by inferred cool sea surface temperatures and reduced surface to deep water temperature gradients. We suggest that deep waters in the eastern Indian Ocean were not directly fed by the Southern or Tethys Oceans. Rather, the more negative delta13C composition of the bottom waters recorded by benthic foraminifera implies the presence and/or active contribution of aged deep waters from the Pacific during this time, at least prior to ~60.2 Ma and subsequent to ~59.0 Ma. The Indian continent, Ninetyeast Ridge, Kerguelen Plateau and Broken Ridge may have played a significant role as submarine barriers to deep water circulation during the Paleocene.