Alterations in the ras oncogene and the p53 tumor suppressor gene in ultraviolet radiation-induced skin carcinogenesis

A rapid increase of the ultraviolet radiation (UVR)-related skin cancer incidence has attracted more and more public attention during the last few decades. Prevention and treatment of UVR-related skin cancer has become an important public health issue in the United States. Recent studies indicate that mutations in ras and/or p53 genes may be involved in UVR-induced skin tumor development but the precise molecular mechanism remains unclear. In this study, alterations of H-ras and p53 genes were investigated in different stages of carcinogenesis in a chronic UVR (solar simulator) exposure-induced Sencar mouse skin carcinogenesis model in order to clarify the role of the alterations of these genes during the skin carcinogenesis process and to further understand the mechanisms by which UVR causes skin cancer.^ Positive ras-p21 staining in cell membranes and cytosol were detected in 18/33 (55%) of squamous cell carcinomas (SCCs), but were not detected in UV-exposed skin, papillomas, or spindle cell tumors (SCTs). Positive staining of the malignant progression marker K13 was found in 17/33 (52%) of SCCs only. A significant positive correlation was observed between the K13 and the ras-p21 expression. Polymerase chain reaction (PCR)-based single strand conformation polymorphism (SSCP) analysis and gene sequencing analysis revealed three point mutations, one (codon 56) in UV-exposed non-tumor bearing skin and the other two (codons 21 and 13) in SCCs. No UV-specific mutation patterns were found.^ Positive p53 nuclear staining was found in 10/37 (27%) of SCCs and 12/24 (50%) of SCTs, but was not detected in normal skin or papillomas. PCR-based SSCP and sequencing analysis revealed eight point mutations in exons 5 and 6 (four in SCTs, two in SCCs, and two in UV-exposed skin) including six C-T or C-A transitions. Four of the mutations occurred at a dipyrimidine (CC) sequence. The pattern of the mutations indicated that the mutagenic lesions were induced by UVR.^ These results indicate that overexpression of ras-p21 in conjunction with aberrant expression of K13 occurred frequently in UVR-induced SCCs in Sencar mouse skin. The point mutation in the H-ras gene appeared to be a rare event in UVR skin carcinogenesis and may not be responsible for overexpression of ras-p21. UVR-induced P53 gene alteration is a frequent event in UVR-induced SCCs and later stage SCT tumors in Sencar mice skin, suggesting the p53 gene mutation plays an important role in skin tumor malignant progression. ^

Regulation of apoptosis during the pathogenesis of non -melanoma skin cancer

Previous studies from our lab have shown distinctive patterns of expression of bcl-2 gene family members in human nonmelanoma skin cancer (NMSC). To further evaluate the significance of these observations and to study the effects of cell death deregulation during skin carcinogenesis, we generated a transgenic mouse model (HK1.bcl-2) using the human keratin 1 promoter to target the expression of a human bcl-2 minigene to the epidermis. Transgenic protein expression was confirmed in all the layers of the epidermis except the stratum corneum using immunohistochemistry. Multifocal epidermal hyperplasia, without associated hyperkeratosis, was observed in newborn HK1.bcl-2 mice. Immunofluorescence staining using monoclonal antibodies specific for a variety of differentiation markers revealed aberrant expression of keratin 6 (K6) in the transgenic epidermis. Epidermal proliferative indexes, assessed by anti-BrdUrd immunofluorescence staining, were similar in control and transgenic newborn mice, but suprabasal proliferating cells were seen within the hyperplastic areas of the transgenic mouse skin. Spontaneous apoptotic indices of the epidermis were similar in both control and HK1.bcl-2 transgenic newborn mice, however, after UV-B irradiation, the number of "sunburn cells" was significantly higher in the control compared to the HK1.bcl-2 transgenic animals.^ Adult HK1.bcl-2 and control littermate mice were used in UV-B and chemical carcinogenesis protocols including DMBA + TPA. UV-B irradiated control and HK1.bcl-2 mice had comparable incidence of tumors than the controls, but the mean latency period was significantly shorter in the HK1.bcl-2 transgenic. Both control and transgenic animals included in chemical carcinogenesis protocols required application of both the initiating (DMBA) and promoting (TPA) agents to develop tumors. The frequency, number, and latency of tumor formation was similar in both groups of animals, however, HK1.bcl-2 mice exhibited a rate of conversion from benign papilloma to carcinoma 2.5 times greater than controls.^ Similar carcinogenesis experiments were performed using newborn mice. HK1.bcl-2 mice treated with UV-B plus TPA have a three fold greater incidence of tumor formation compared to controls littermates. HK1.bcl-2 transgenic animals also exhibited a shorter latency for papilloma formation when treated with DMBA plus TPA.^ HK1.bcl-2/v-Ha-ras double transgenic mice shared phenotypic features of both HK1.v-Ha-ras and HK1.bcl-2 transgenic mice, and exhibited focal areas of augmented hyperplasia. These double transgenic mice were susceptible to tumor formation by treatment with TPA alone.^ Cultures of primary keratinocytes were established from control, HK1.bcl-2, HK1.Ha-ras, and HK1.bcl-2/v-Ha-ras newborn mice. Cell viability was determined after exposure of the cells to UV-B irradiation, DMBA, TPA, or TGF-$\beta$1. Internucleosomal DNA fragmentation ("ladders") and morphological cellular changes compatible with apoptotic cell death were observed after the application of all these agents. HK1.bcl-2 keratinocytes were resistant to cell death induction by all of these agents except TGF-$\beta$1. HK1.Ha-ras cells had a higher spontaneous rate of cell death which could be compensated by co-expression of bcl-2.^ These findings suggest that bcl-2 dependent cell death suppression may be an important component of multistep skin carcinogenesis. ^

Fas ligand and soluble Fas -opposing molecules in melanoma lung metastasis

Deregulation of apoptotic cell death can result in aberrant accumulation of cells and increased tumor incidence. Fas (CD95) and Fas ligand (FasL) are a receptor-ligand pair whose activation induces apoptosis in many cell types. Previously, we demonstrated that low metastatic, Fas+ K1735-P murine melanoma cells spontaneously metastasize to the lung following orthotopic injection into FasL-deficient (gld) mice compared to wild-type (wt) controls. We further demonstrated that the expression of the Fas antagonist soluble Fas (sFas) directly correlates with disease stage in patients with melanoma, breast, and colon cancer. These findings document a role for host-derived FasL, in the control of metastatic disease and suggest a role for tumor-associated sFas in acquiring metastatic potential. To directly test whether FasL expressed on lymphocytes or on lung stromal cells restricts metastasis, bone marrow chimeras were generated between C3H wt and C3H gld mice. Chimeric animals were injected subcutaneously with 5 × 105 K1735-P and the incidence and number of spontaneous lung metastases scored. The data show that wt mice receiving gld marrow had a greater number of lung metastases (median 9.5, range 2–31) than gld mice reconstituted with wt marrow (median 1, range 0–31; p < 0.016). Interestingly, both groups had fewer metastases compared to gld controls (median 18.5, range 0–46) but more than wt controls (median 2, range 0–7). These observations provide the first evidence that both hematopoietic- and nonhematopoietic-host derived FasL, are important in the control of melanoma metastasis to the lung. To directly test whether tumor-associated sFas expression can enhance metastasis, K1735-P cells were transfected with three isoforms of sFas (Exo4Del, Exo6Del, and Exo3, 4, 6Del). RT-PCR and ELISA analysis confirmed the expression of sFas RNA and protein respectively. Following intravenous injection of 5 × 104 cells, sFas transfected cells formed significantly more experimental lung metastases [Exo6Del clone 3 (median 22, range 0–36), Exo6Del clone 7 (median 31, range 4–50), Exo3, 4, 6Del (median 22.5, range 13–48)] compared to vector control cells (median 6.5, range 3–29). Together, these data provide the first evidence that sFas is sufficient to enhance the metastatic potential of Fas+ melanoma cells. ^

Signaling pathways in the transcriptional and post-translational regulation of the human glutathione S -transferase P1 gene

The human glutathione S-transferase P1 (GSTP1) protein is an endogenous inhibitor of c-jun N-terminal kinases (JNKs) and an important phase II detoxification enzyme. ^ Recent identification of a cAMP response element (CRE) in the 5 ′-region of the human GSTP1 gene and several putative phosphorylation sites for the Ser/Thr protein kinases, including, cAMP-dependent protein kinases (PKAs), protein kinases C (PKCs), and JNKs in the GSTP1 protein raised the possibility that signaling pathways may play an important role in the transcriptional and post-translational regulation of GSTP1 gene. This study examined (a) whether the signaling pathway mediated by CAMP, via the GSTP1 CRE, is involved in the transcriptional regulation of the GSTP1 gene, (b) whether signaling pathways mediated by the Ser/Thr protein kinases (PKAs, PKCs, and JNKs) induce post-translational modification, viz. phosphorylation of the GSTP1 protein, and (c) whether such phosphorylation of the GSTP1 protein alters its functions in metabolism and in JNK signaling. ^ The first major finding in this study is the establishment of the human GSTP1 gene as a novel CAMP responsive gene in which transcription is activated via an interaction between PKA activated CRE binding protein-1 (CREB-1) and the CRE in the 5′-regulatory region. ^ The second major finding in this study is the observation that the GSTP1 protein undergoes phosphorylation and functionally activated by second messenger-activated protein kinases, PKA and PKC, in tumor cells with activated signaling pathways. Following phosphorylation by PKA or PKC, the catalytic activity of the GSTP1 protein was significantly enhanced, as indicated by a decrease in its Km (2- to 3.6-fold) and an increase in Kcat/ Km (1.6- to 2.5-fold) for glutathione. Given the frequent over-expression of GSTP1 and the aberrant PKA/PKC signaling cascade observed in tumors, these findings suggest that phosphorylation of GSTP1 may contribute to the malignant progression and drug-resistant phenotype of these tumors. ^ The third major finding in this study is that the GSTP1 protein, an inhibitor of JNKs, undergoes significant phosphorylation in tumor cells with activated JNK signaling pathway and in those under oxidative stress. Following phosphorylation by JNK, the ability of GSTP1 to inhibit JNK downstream function, i.e. c-jun phosphorylation, was significantly enhanced, suggesting a feedback mechanism of regulation of JNK-mediated cellular signaling. (Abstract shortened by UMI.) ^

Pathways leading to apoptosis resistance in a murine B -cell lymphoma cell system

The aberrant activation of signal transduction pathways has long been linked to uncontrolled cell proliferation and the development of cancer. The activity of one such signaling module, the Mitogen-Activated Protein Kinase (MAPK) pathway, has been implicated in several cancer types including pancreatic, breast, colon, and lymphoid malignancies. Interestingly, the activation of MAP-Kinase-Kinase-Kinase proteins often leads to the additional activation of NF-κB, a transcription factor that acts as a cell survival signal through its control of antiapoptotic genes. We have investigated the role of a specific dimer form of the NF-κB transcription factor family, NF-κB1 (p50) homodimers, in its control of the proto-oncogene, Bcl-2, and we have identified the MEK/ERK (MAPK) signaling cascade as a mediator of NF-κB1 activity. ^ Two murine B cell lymphoma cell lines were used for these studies: LY-as, an apoptosis proficient line with low Bcl-2 protein expression and no nuclear NF-κB activity, and LY-ar, a nonapoptotic line with constitutive p50 homodimer activity and 30 times more Bcl-2 protein expression than LY-as. Experiments modulating p50 activity correlated the activation of p50 homodimers with Bcl-2 expression and additional gel shift experiments demonstrated that the Bcl-2 P1 promoter had NF-κB sites with which recombinant p50 was able to interact. In vitro transcription revealed that p50 enhanced the production of transcripts derived from the Bcl-2 P1 promoter. These data strongly suggest that Bcl-2 is a target gene for p50-mediated transcription and suggest that the activation of p50 homodimers contributes to the expression of Bcl-2 observed in LY-ar cells. ^ Studies of upstream MAPK pathways that could influence NF-κB activity demonstrated that LY-ar cells had phosphorylated ERK proteins while LY-as cells did not. Treatment of LY-ar cells with the MEK inhibitors PD 98059, U0126, and PD 184352 led to a loss of phosphorylated ERK, a reversal of nuclear p50 homodimer DNA binding, and a decrease in the amount of Bcl-2 protein expression. Similarly, the activation of the MEK/ERK pathway in LY-as cells by phorbol ester led to Bcl-2 expression that could be blocked by PD 98059. Furthermore, treatment of LY-ar cells with TNFα, an IKK activator, did not change the suppressive effect of PD 98059 on p50 homodimer activity, suggesting an IKK-independent pathway for p50 homodimer activation. Lastly, all three MEK inhibitors sensitized LY-ar cells to radiation-induced apoptosis. ^ These data indicate that the activation of the MEK/ERK MAP-Kinase signaling pathway acts upstream of p50 homodimer activation and Bcl-2 expression in this B cell lymphoma cell system and suggest that the activation of MEK/ERK may be a key step in the progression of lymphoma to advanced-staged disease. Other researchers have used MEK inhibitors to inhibit cell growth and sensitize a number of tumors to chemotherapies. In light of our data, MEK inhibitors may additionally be useful clinically to radiosensitize cancers of lymphoid origin. ^

Using K5 Myc transgenic mice to delineate *Myc's apoptotic and tumorigenic pathways

The c-myc oncogene has the unusual ability to induce proliferation and apoptosis. Transgenic mice have been generated in which the expression of Myc is under the control of an epithelial-specific keratin 5 (K5) promoter. These mice have increased levels of proliferation and p53-dependent apoptosis, and are predisposed to developing spontaneous tumors in epithelial tissues. In this study, various knockout mice were bred to K5 Myc transgenic mice to identify factors involved in the aberrant apoptosis, hyperproliferation, and spontaneous tumorigenesis present in these mice. Consistent with in vitro studies, Myc-induced, p53-dependent apoptosis in transgenic epidermis was found to be partially dependent on p19ARF, a p53 regulator that inhibits mdm2. Additionally, the rate of tumorigenesis was increased when p19ARF was absent in Myc transgenic mice. Consistent with previous reports that some E2F family members may function as tumor suppressors, inactivation of either E2f1 or E2f2 was found to accelerate tumor development in the K5 Myc transgenic mice. Acceleration of tumorigenesis in the absence of E2F1 occurred despite the fact that apoptotic levels were increased in transgenic tissue and tumors null for E2f1 , whereas hyperproliferation was unaffected. In contrast, inactivation of E2f2 was found to increase hyperproliferation in the K5 Myc transgenic mice, while having no effect on apoptosis. The lack of E2f1 in the Myc transgenic mice increased the expression of several p53 transcription target genes, which may explain the increased apoptosis in these mice. In transgenic epidermis, p53 is phosphorylated at serine 18, a site of phosphorylation by ATM. Inactivation of ATM in K5 Myc transgenic mice impaired Myc-induced apoptosis, identifying ATM as having an important role in Myc-induced apoptosis. Moreover, the absence of ATM accelerates tumorigenesis in K5-expressing tissues. However, p53 accumulation and phosphorylation at serine 18 induced by Myc occurs independent of ATM. Therefore, another activity of ATM appears to be important for Myc-induced apoptosis. These findings show that acceleration of tumorigenesis in K5 Myc transgenic mice, as in the case of p53, p19ARF, E2F1, E2F2, and ATM absence, does not necessarily correlate with suppression of Myc-induced apoptosis, as seen only when p53, p19ARF or ATM was absent. ^

Silicoflagellates of early Paleocene to early Miocene sediments of the subantarctic South Atlantic

Nearly complete Paleogene sedimentary sequences were recovered by Leg 114 to the subantarctic South Atlantic. Silicoflagellate assemblages from the Paleogene and immediately overlying lower Neogene from Sites 698 (Northeast Georgia Rise), 700 (East Georgia Basin), 702 (Islas Orcadas Rise), and 703 (Meteor Rise) were examined. The described assemblage from Hole 700B represents the most complete yet described from the Paleocene, encompassing planktonic foraminifer Zones Plb (upper part) through P4 and Subchrons C25N to C23N. All lower Eocene sediments are barren as a result of diagenesis, except for a single sample from Hole 698A. Middle Eocene silicoflagellates described from Hole 702B range in age from early middle Eocene (P10) to late Eocene (PI5), with correlations to Subchrons C21N to C18N. Hole 703A contains late Eocene through early Miocene assemblages, with paleomagnetic control from Subchrons C16R to C6AAN. Leg 114 biosiliceous sequences contain exceptionally diverse assemblages of silicoflagellates. Approximately 155 species and separate morphotypes are described from the Paleogene and earliest Neogene. New taxa described from Leg 114 sediments include Bachmannocena vetula n. sp., Corbisema animoparallela n. sp., Corbisema camara n. sp., Corbisema constricta spinosa n. subsp., Corbisema delicata n. sp., Corbisema hastata aha n. subsp., Corbisema praedelicata n. sp., Corbisema scapana n. sp., Corbisema triacantha lepidospinosa n. subsp., Dictyocha deflandreifurtivia n. subsp., Naviculopsis biapiculata nodulifera n. subsp., Naviculopsis cruciata n. sp., Naviculopsis pandalata n. sp., Naviculopsis primativa n. sp., and Naviculopsis trispinosa eminula n. subsp. Taxonomic revisions were made to the following taxa: Corbisema constricta constricta emended, Corbisema disymmetrica crenulata n. comb., Corbisema jerseyensis emended, and Distephanus antarcticus n. comb. Silicoflagellate assemblages from the Paleogene and earliest Neogene of Holes 698A, 699A, 700B, 702B, and 703A are the basis of a silicoflagellate zonation spanning the interval from 63.2 to 22.25 Ma. Silicoflagellate zones recognized in this interval include the Corbisema hastata hastata Zone, Corbisema hastata aha Zone, Dictyocha precarentis Zone, Naviculopsis constricta Zone, Naviculopsis foliacea Zone, Bachmannocena vetula Zone, Dictyocha grandis Zone, Naviculopsis pandalata Zone, Naviculopsis constricta-Bachmannocena paulschulzii Zone, Bachmannocena paulschulzii Zone, Naviculopsis trispinosa Zone with subzones a and b, Corbisema archangelskiana Zone, Naviculopsis biapiculata Zone, Distephanus raupii Zone, Distephanus raupii-Corbisema triacantha Zone, and Corbisema triacantha mediana Zone.

Abundances and contents of major silicoflagellates from IODP Holes of Exp302

The silicoflagellate taxa obtained in IODP Expedition 302 (ACEX) were identified and counted in order to establish the silicoflagellate biostratigraphy in the central Arctic Ocean. These microfossils in the ACEX samples were preserved in the Lithology Units 1/6 and 2, which are dark silty clay and biosiliceous ooze, respectively. The silicoflagellate skeletons in the ACEX samples are assigned to 56 taxa. Seven taxa were described as new species, which were abundant in Lithology Unit 2. Comparison with several study cases outside the Eocene Arctic Ocean suggested that the silicoflagellate assemblages in ACEX were unique in Lithology Unit 2. The dominance of silicoflagellate taxa varied throughout the lithological section. Based on the cluster analysis by Morishita similarity index C(Lambda), the silicoflagellate assemblageswere divided into nine assemblage groups. The silicoflagellate datum event of the first occurrence of Corbisema hexacantha in the lower part of Lithology Unit 1/6 is regarded. Based on the datum events for silicoflagellate and palynomorphs, the assigned epoch of Lithology Units 1/6 and 2 is the middle Eocene.

Silicoflagellates and ebridians from ODP Leg 138 holes

The biostratigraphic distribution and abundance of middle Miocene to Pleistocene silicoflagellates is documented from Ocean Drilling Program (ODP) Leg 138 Holes 844B, 847B, 848B, 849B, 850B, 85 IB, 852B, and 854B from the eastern Equatorial Pacific Ocean. The silicoflagellates were generally abundant and well preserved and frequently exhibited an unusually large range of variation. The upper Miocene of near-equatorial sites includes an assemblage of Bachmannocena diodon nodosa, which includes a bridge across the width of the basal ring. Stratigraphically below this, at sites within 5° of the equator is a lengthy interval of specimens of Distephanus speculum tenuis, which have a fragile apical structure. Both the intervals of Bachmannocena diodon nodosa plexus and Distephanus speculum tenuis are biostratigraphically useful within 5° of the equator, but are less useful beyond that. An unusual range of variation also is observed for Dictyocha in the Pliocene sediments at about the point where D. perlaevis and D. messanensis appear in the geologic record. This variation may be explained by hybridization between diverging species.

Silicoflagellate abundances in sediments of ODP Leg 207 sites

The Ocean Drilling Program (ODP) drilled at five sites in the western Atlantic Ocean during Leg 207. The objective of the drilling was to recover samples from the shallow buried Cretaceous and Paleocene sediments on the Demerara Rise off Suriname, South America. These sediments are being studied for a number of paleoceanographic studies of the low-latitude Atlantic off the coast of Suriname (this volume). For this report two sites, Sites 1257 and 1258, were selected for silicoflagellate study because shipboard results suggested these two sites as the only ones with siliceous microfossils of Paleocene-Eocene age. The Demarara Rise is a predominant submarine plateau located off the coast of Suriname and French Guyana. This plateau stretches 380 km along the coast and is 220 km wide. The depth to seafloor along the depth transect drilled during ODP Leg 207 ranges from 1000 to 4500 m, but most of the remainder of the plateau lies in shallow water of 700 m. Much of this area is covered with 2-3 km of sediments. The Demerara Rise is built on rifted Precambrian continental crust. The plateau was one of the last places to be in contact with West Africa during the opening of the Atlantic Ocean (see Shipboard Scientific Party, 2004). Site 1257 (9°27'N, 54°20'W; water depth = 2951 m) is located on a terrace on the northwestern Demerara Rise ~400 km from Suriname. This is the second deepest water depth location drilled during Leg 207. Sediments from this area range in age from Miocene to Albian. This area is part of the transform fault that separated from Central America and western Africa. Three holes were drilled at Site 1257. Site 1258 (9°26'N, 54°43'W; water depth = 3192 m) is located on the western slope of the Demerara Rise ~380 km north of Suriname. This site is the distal and deepest site of the paleoceanographic depth transect drilled across Demerara Rise during Leg 207. The area is located on a ridge of Paleocene sediments cropping out on the seafloor. Three holes were drilled at Site 1258, but only one is studied.

Silicoflagellate abundance in ODP Holes 183-1138A and 183-1140A

The biostratigraphic distribution and abundance of lower Oligocene and Miocene to Pleistocene silicoflagellates are documented from Ocean Drilling Program Leg 183 Holes 1138A and 1140A, on the Kerguelen Plateau. The Distephanus speculum speculum forma pseudofibula plexus is found in the upper Miocene in Hole 1138A, but other important biostratigraphic markers are not available. Diversity and abundance of silicoflagellates vary considerably in Hole 1138A, with silicoflagellates more abundant in the Pliocene and Pleistocene and some intervals of the Miocene barren of silicoflagellates or containing only limited numbers of specimens. The silicoflagellates of Hole 1140A include a new skeletal morphology, described here as Distephanus speculum speculum forma cylindrus. Silicoflagellates were generally abundant throughout the lower and middle Miocene in Hole 1140A.

Calcareous nannofossil during the Paleocene Eocene Thermal Maximum in sediments of Walvish Ridge

Compositions and abundances of calcareous nannofossil taxa have been determined in a ca 170 kyrs long time interval across the Paleocene/Eocene boundary at 1-cm to 10-cm resolution from two ODP Sites (1262, 1263) drilled along the flank of the Walvis Ridge in the South Atlantic. The results are compared to published data from ODP Site 690 in the Weddell Sea. The assemblages underwent rapid evolution over a 74 kyrs period, indicating stressed, unstable and/or extreme photic zone environments during the PETM hyperthermal. This rapid evolution, which created 5 distinct stratigraphic horizons, is consistent with the restricted brief occurrences of malformed and/or weakly calcified morphotypes. The production of these aberrant morphotypes is possibly caused by major global scale changes in carbon cycling in the ocean-atmosphere system, affecting also photic zone environments. No marked paleoecologically induced changes are observed in abundances of the genera Discoaster, Fasciculithus and Sphenolithus at the Walvis Ridge sites. Surprisingly, there is no significant correlation in abundance between these three genera, presumed to have had a similar paleoecological preference for warm and oligotrophic conditions.

Siliceous microfossil assemblage and accumulation rate variations across the Middle Eocene Climatic Optimum at ODP Site 171-1051

The Middle Eocene Climatic Optimum (MECO; ~ 40 million years ago [Ma]) is one of the most prominent transient global warming events in the Paleogene. Although the event is well documented in geochemical and isotopic proxy records at many locations, the marine biotic response to the MECO remains poorly constrained. We present new high-resolution, quantitative records of siliceous microplankton assemblages from the MECO interval of Ocean Drilling Program (ODP) Site 1051 in the subtropical western North Atlantic Ocean, which are interpreted in the context of published foraminiferal and bulk carbonate stable isotope (d18O and d13C) records. High diatom, radiolarian and silicoflagellate accumulation rates between 40.5 and 40.0 Ma are interpreted to reflect an ~ 500 thousand year (kyr) interval of increased nutrient supply and resultant surface-water eutrophication that was associated with elevated sea-surface temperatures during the prolonged onset of the MECO. Relatively low pelagic siliceous phytoplankton sedimentation accompanied the peak MECO warming interval and the termination of the MECO during an ~ 70 kyr interval centered at ~ 40.0 Ma. Following the termination of the MECO, an ~ 200-kyr episode of increased siliceous plankton abundance indicates enhanced nutrient levels between ~ 39.9 and 39.7 Ma. Throughout the Site 1051 record, abundance and accumulation rate fluctuations in neritic diatom taxa are similar to the trends observed in pelagic taxa, implying either similar controls on diatom production in the neritic and pelagic zones of the western North Atlantic or fluctuations in sea level and/or shelf accommodation on the North American continental margin to the west of Site 1051. These results, combined with published records based on multiple proxies, indicate a geographically diverse pattern of surface ocean primary production changes across the MECO. Notably, however, increased biosiliceous accumulation is recorded at both ODP Sites 1051 and 748 (Southern Ocean) in response to MECO warming. This may suggest that increased biosiliceous sediment accumulation, if indeed a widespread phenomenon, resulted from higher continental silicate weathering rates and an increase in silicic acid supply to the oceans over several 100 kyr during the MECO.

Neogene silicoflaggelates in ODP Leg 104 holes

A quantative study was made of silicoflagellates recovered from Sites 642 (lower Miocene-upper Pliocene), 643 (lower Miocene-upper Miocene), and 644 (upper Pliocene-Quaternary) on the Voring Plateau. Although disconformities are present in these sequences, they represent a much more complete record of the Neogene than was recovered previously in the Norwegian Sea by DSDP Leg 38. Silicoflagellates are rare or absent for glacial sequences younger than 2.65 Ma, and generally sparse and poorly preserved in the lower upper Pliocene and upper Miocene. Lower and middle Miocene assemblages are diverse and generally well preserved. Temporal changes in the silicoflagellate assemblage are indicative of major paleoceanographic changes in the Norwegian Sea. A regional zonation for the Neogene of the Norwegian Sea is proposed, consisting of eleven zones: Naviculopsis lata Zone, N. quadrata Zone (emended), N. ponticula Zone (emended), Distephanus speculum hemisphaericus Zone (new), Caryocha ernestinae Zone (new), Bachmannocena circulus var. apiculata/Caryocha Zone (new), Distephanus crux scutulatus Zone (new), Bachmannocena diodon nodosa Zone (new), Distephanus boliviensis Zone (new), Ds. jimlingii Zone (elevated from subzonal to zonal status) with Subzones a and b (new), and Ds. speculum Zone (new). The ranges and abundances of over 100 species and morphotypes are tabulated.