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Perfluorinated alkyl acids (PFAAs) are persistent in marine biota and are toxic to many species, including marine mammals. We measured the concentrations of 15 PFAAs in liver and kidney samples of 16 species of stranded cetaceans from Hawai'i and other tropical North Pacific regions utilizing high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). Eleven PFAAs in liver and nine PFAAs in kidney were detected, including substantial perfluorooctanesulfonate (PFOS) and perfluoroundecanoic acid (PFUnA). Regression models indicated that phylogenetic family and age class significantly influenced concentrations of certain PFAAs. PFAAs can activate transcription factor peroxisome proliferator-activated receptor alpha (PPARα), which induces transcription of cytochrome P450 4A (CYP4A). Relative expression of PPARα and CYP4A mRNA was quantified using real-time PCR (qPCR) and CYP4A protein expression, using Western blot and then compared to PFAA concentrations in liver and kidney. Concentrations of four PFAA congeners, summation of perfluoroalkyl carboxylic acids (ΣPFCAs), and ΣPFAAs correlated significantly with PPARα mRNA expression and CYP4A protein expression in kidney, suggesting either may be biomarkers of PFAA exposure in cetaceans. This is the first study to quantify PFAAs in marine mammals from this region and the first observation of a direct relationship between PFAA exposure and PPARα and CYP4A expression in cetaceans.

Pelagic Pacific sea turtles eat relatively large quantities of plastic (median 5 g in gut). Using Fourier transform infrared spectroscopy, we identified the polymers ingested by 37 olive ridley, 9 green, and 4 loggerhead turtles caught as bycatch in Hawaii- and American Samoa-based longline fisheries. Unidentifiable samples were analyzed using high-temperature size exclusion chromatography with multiple detectors and/or X-ray photoelectron spectroscopy. Regardless of species differences in dive depths and foraging strategies, ingested plastics were primarily low-density, floating polymers (51% low-density polyethylene (LDPE), 26% polypropylene (PP), 10% unknown polyethylene (PE), and 5% high-density PE collectively). Albeit not statistically significant, deeper diving and deeper captured olive ridley turtles ate proportionally more plastics expected to sink (3.9%) than intermediate-diving green (1.2%) and shallow-diving loggerhead (0.3%) turtles. Spatial, but no sex, size, year, or hook depth differences were observed in polymer composition. LDPE and PP, some of the most produced and least recycled polymers worldwide, account for the largest percentage of plastic eaten by sea turtles in this region. These novel data inform managers about the threat of plastic ingestion to sea turtles and may motivate development of more environmentally friendly practices for plastic production, use, and waste management.

Elevated levels of persistent organic pollutants (POPs) have been reported in tropical Pacific Island cetaceans and their environment. In addition, recent health concerns in cetacean populations have warranted investigation into potential physiological effects from POP exposure for this region. Cytochrome P450 1A1 (CYP1A1) is a candidate for examining such effects. This well-studied biomarker of exposure and effect was examined in stranded cetacean liver using immunoblot (n = 39, 16 species) and blubber using immunohistochemistry (n = 23, 10 species). Paired tissue samples allowed for CYP1A1 comparisons not only between species but also within each individual animal to examine differences between tissue types. Liver CYP1A1 expression correlated positively and significantly with blubber concentrations of all POP categories (n = 39, p 0.100). Among Stenella species, liver CYP1A1 tissue expression was correlated negatively with the sum of all blubber layer endothelial cell CYP1A1 expression (n = 14, p = 0.049). Overall, elevated expression of liver CYP1A1 confirms its use as a biomarker of POP exposure to cetaceans stranded in the tropical Pacific basin. Environ Toxicol Chem 2015;34:1989–1995. © 2015 SETAC

The viral genus Morbillivirus and the bacterial genus Brucella have emerged as important groups of pathogens that are known to affect cetacean health on a global scale, but neither pathogen has previously been reported from endangered sperm whales (Physeter macrocephalus). A female neonate sperm whale stranded alive and died near Laie on the island of Oahu, Hawaii, US, in May of 2011. Congestion of the cerebrum and enlarged lymph nodes were noted on the gross necropsy. Microscopic findings included lymphoid depletion, chronic meningitis, and pneumonia, suggesting an in utero infection. Cerebrum, lung, umbilicus, and select lymph nodes (tracheobronchial and mediastinal) were positive for Brucella by PCR. Brucella sp. was also cultured from the cerebrum and from mediastinal and tracheobronchial lymph nodes. Twelve different tissues were screened for Morbillivirus by reverse-transcriptase (RT)-PCR and select tissues by immunohistochemistry, but only the tracheobronchial lymph node and spleen were positive by RT-PCR. Pathologic findings observed were likely a result of Brucella, but Morbillivirus may have played a key role in immune suppression of the mother and calf. The in utero infection in this individual strongly supports vertical transmission of both pathogens.

Polymer identification of plastic marine debris can help identify its sources, degradation, and fate. We optimized and validated a fast, simple, and accessible technique, attenuated total reflectance Fourier transform infrared spectroscopy (ATR FT-IR), to identify polymers contained in plastic ingested by sea turtles. Spectra of consumer good items with known resin identification codes #1-6 and several #7 plastics were compared to standard and raw manufactured polymers. High temperature size exclusion chromatography measurements confirmed ATR FT-IR could differentiate these polymers. High-density (HDPE) and low-density polyethylene (LDPE) discrimination is challenging but a clear step-by-step guide is provided that identified 78% of ingested PE samples. The optimal cleaning methods consisted of wiping ingested pieces with water or cutting. Of 828 ingested plastics pieces from 50 Pacific sea turtles, 96% were identified by ATR FT-IR as HDPE, LDPE, unknown PE, polypropylene (PP), PE and PP mixtures, polystyrene, polyvinyl chloride, and nylon.

Persistent organic pollutants such as halogenated aromatic hydrocarbons (HAHs) biomagnify in food webs and accumulate to high concentrations in top predators like odontocete cetaceans (toothed whales). The most toxic HAHs are the 2,3,7,8-substituted halogenated dibenzo-p-dioxins and furans, and nonortho-substituted polychlorinated biphenyls (PCBs), which exert their effects via the aryl hydrocarbon receptor (AHR). Understanding the impact of HAHs in wildlife is limited by the lack of taxon-specific information about the relative potencies of toxicologically important congeners. To assess whether Toxic Equivalency Factors (TEFs) determined in rodents are predictive of HAH relative potencies in a cetacean, we used beluga and mouse AHRs expressed in vitro from cloned cDNAs to measure the relative AHRbinding affinities of ten HAHs from five different structural classes. The rank order of mean IC50s for competitive binding to beluga AHR was: TCDD < TCDF < PCB-126 < PCB-169 < PCB-77 < PCB-81 ≪ PCB156 ∼ PCB-128 < PCB-105 < PCB-118. The rank order of mean IC50s for binding to the mouse AHR was TCDD < TCDF < PCB-126 < PCB-169 < PCB-81 < PCB-77 < PCB-156 � PCB-128 ∼ PCB-105 ∼ PCB-118. Ki values for binding of HAHs to beluga and mouse AHRs were highly correlated (r 2 = 0.96). Comparison of Ki values suggested that the beluga AHR had a higher affinity than the mouse AHR for most of the HAHs tested, consistent with the ∼2-fold higher [3H]TCDD binding affinity determined previously. These results are consistent with the World Health Organization mammalian TEFs for non- and mono-ortho PCB congeners. The comparatively high HAH binding affinities of the beluga AHR relative to those of an AHR from a dioxin-responsive mouse suggests that beluga, and perhaps cetaceans in general, may be particularly sensitive to the toxic effects of AHR agonists. Further study is warranted in order to more fully address this important question affecting protected and endangered species.

Cetacean morbillivirus (CeMV) is a causative factor in epizootics that have resulted in thousands of deaths throughout the Atlantic and Mediterranean since 1987, but less is known of its presence and significance in the Pacific. The first case of CeMV reported in Hawai'i was in a Longman's beaked whale that stranded in 2010. The initial CeMV sequence from this individual indicated the possibility of a novel strain. To address this, archived samples from cetaceans that stranded in Hawai'i between 1997 and 2014 were screened for CeMV. The beaked whale morbillivirus (BWMV) was detected in 15 individuals representing 12 different species (24% of Code 1 and 2 stranded cetaceans). The earliest detected case was a humpback whale that stranded in 1998. Sequence comparisons of a 2.2 kb sequence spanning the phosphoprotein (P) and nucleocapsid (N) genes strongly suggest that the BWMV represents a novel strain of CeMV present in Hawai'i and the Central Pacific. In contrast to recently reported isolates from Brazil and Australia that may represent a distinct clade, BWMV appears to be more closely related to known strains of CeMV (dolphin morbillivirus; porpoise morbillivirus; and pilot whale morbillivirus). Detection rates with repeat sampling of positive lymph nodes were between 2 and 61%, illustrating the extreme heterogeneity that can occur in affected tissues. Taken together, these results suggest that BWMV may be common and established in Hawaiian cetacean populations. BWMV will be important for understanding CeMV and health threats in the relatively understudied cetaceans of the Pacific.

The impacts of anthropogenic contaminants on marine ecosystems are a concern worldwide. Anthropogenic activities can enrich trace elements in marine biota to concentrations that may negatively impact organism health. Exposure to elevated concentrations of trace elements is considered a contributing factor in marine mammal population declines. Hawai'i is an increasingly important geographic location for global monitoring, yet trace element concentrations have not been quantified in Hawaiian cetaceans, and there is little trace element data for Pacific cetaceans. This study measured trace elements (Cr, Mn, Cu, Zn, As, Se, Sr, Cd, Sn, Hg, and Pb) in liver of 16 species of cetaceans that stranded on U.S. Pacific Islands from 1997 to 2013, using high resolution inductively coupled plasma mass spectrometry (HR-ICP-MS) (n = 31), and direct mercury analysis atomic absorption spectrometry (DMA-AAS) (n = 43). Concentration ranges (μg/g wet mass fraction) for non-essential trace elements, such as Cd (0.0031-58.93) and Hg (0.0062-1571.75) were much greater than essential trace elements, such as Mn (0.590-17.31) and Zn (14.72-245.38). Differences were found among age classes in Cu, Zn, Hg, and Se concentrations. The highest concentrations of Se, Cd, Sn, Hg, and Pb were found in one adult female false killer whale (Pseudorca crassidens) at concentrations that are known to affect health in marine mammals. The results of this study establish initial trace element concentration ranges for Pacific cetaceans in the Hawaiian Islands region, provide insights into contaminant exposure of these marine mammals, and contribute to a greater understanding of anthropogenic impacts in the Pacific Ocean.

Polycyclic aromatic hydrocarbons (PAH) are common environmental pollutants that suppress the immune system in part by inducing pro/pre-B cell apoptosis. The PAH-induced death signaling pathway resembles the signaling cascade activated during clonal deletion and modeled by B cell receptor cross-linking or by dexamethasone exposure of immature surface Ig+ B cells in that apoptosis is mediated by NF-κB down-regulation. Because a PAH-induced, clonally nonrestricted deletion of B cells would have important implications for B cell repertoire development, the nature of the PAH-induced intracellular death signal was studied further. Particular emphasis was placed on the roles of growth arrest and c-Myc, p27Kip1, and p21WAF1 expression, because all of these elements contribute to clonal deletion. As in clonal deletion models, and as predicted by the down-regulation of NF-κB, PAH-induced death of pro/pre-B cells was at least partially dependent on c-Myc down-regulation. Furthermore, whereas dexamethasone induced a G0/G1 cell cycle arrest, PAH had no effect on pro/pre-B cell growth, indicating that growth arrest and apoptosis occur by separable signaling pathways in this early phase of B cell development. Finally, in contrast to clonal deletion, PAH-induced pro/pre-B cell death was not dependent on p27Kip1 or p21WAF1 up-regulation but did coincide with p53 induction. These results distinguish the PAH-induced apoptosis pathway from that activated during clonal deletion and indicate that signaling cascades leading to growth arrest and/or apoptosis in pro/pre-B cells differ from those active at later B cell developmental stages.

The role of peroxisome proliferator-activated receptor gamma (PPARgamma) in adipocyte physiology has been exploited for the treatment of diabetes. The expression of PPARgamma in lymphoid organs and its modulation of macrophage inflammatory responses, T cell proliferation and cytokine production, and B cell proliferation also implicate it in immune regulation. Despite significant human exposure to PPARgamma agonists, little is known about the consequences of PPARgamma activation in the developing immune system. Here, well-characterized models of B lymphopoiesis were used to investigate the effects of PPARgamma ligands on nontransformed pro/pre-B (BU-11) and transformed immature B (WEHI-231) cell development. Treatment of BU-11, WEHI-231, or primary bone marrow B cells with PPARgamma agonists (ciglitazone and GW347845X) resulted in rapid apoptosis. A role for PPARgamma and its dimerization partner, retinoid X receptor (RXR)alpha, in death signaling was supported by 1) the expression of RXRalpha mRNA and cytosolic PPARgamma protein, 2) agonist-induced binding of PPARgamma to a PPRE, and 3) synergistic increases in apoptosis following cotreatment with PPARgamma agonists and 9-cis-retinoic acid, an RXRalpha agonist. PPARgamma agonists activated NF-kappaB (p50, Rel A, c-Rel) binding to the upstream kappaB regulatory element site of c-myc. Only doses of agonists that induced apoptosis stimulated NF-kappaB-DNA binding. Cotreatment with 9-cis-retinoic acid and PPARgamma agonists decreased the dose required to activate NF-kappaB. These data suggest that activation of PPARgamma-RXR initiates a potent apoptotic signaling cascade in B cells, potentially through NF-kappaB activation. These results have implications for the nominal role of the PPARgamma in B cell development and for the use of PPARgamma agonists as immunomodulatory therapeutics.

Elevated levels of persistent organic pollutants (POPs) have been reported in tropical Pacific Island cetaceans and their environment. In addition, recent health concerns in cetacean populations have warranted investigation into potential physiological effects from POP exposure for this region. Cytochrome P450 1A1 (CYP1A1) is a candidate for examining such effects. This well-studied biomarker of exposure and effect was examined in stranded cetacean liver using immunoblot (n = 39, 16 species) and blubber using immunohistochemistry (n = 23, 10 species). Paired tissue samples allowed for CYP1A1 comparisons not only between species but also within each individual animal to examine differences between tissue types. Liver CYP1A1 expression correlated positively and significantly with blubber concentrations of all POP categories (n = 39, p0.050) except octachlorostyrene and pentachlorobenzene (p0.100). Among Stenella species, liver CYP1A1 tissue expression was correlated negatively with the sum of all blubber layer endothelial cell CYP1A1 expression (n = 14, p = 0.049). Overall, elevated expression of liver CYP1A1 confirms its use as a biomarker of POP exposure to cetaceans stranded in the tropical Pacific basin.

Odontocetes (toothed whales) are considered sentinel species in the marine environment because of their high trophic position, long life spans, and blubber that accumulates lipophilic contaminants. Cytochrome P4501A1 (CYP1A1) is a biomarker of exposure and molecular effects of certain persistent organic pollutants. Immunohistochemistry was used to visualize CYP1A1 expression in blubber biopsies collected by non-lethal sampling methods from 10 species of free-ranging Hawaiian odontocetes: short-finned pilot whale, melon-headed whale, pygmy killer whale, common bottlenose dolphin, rough-toothed dolphin, pantropical spotted dolphin, Blainville’s beaked whale, Cuvier’s beaked whale, sperm whale, and endangered main Hawaiian Islands insular false killer whale. Significantly higher levels of CYP1A1 were observed in false killer whales and rough-toothed dolphins compared to melon-headed whales, and in general, trophic position appears to influence CYP1A1 expression patterns in particular species groups. No significant differences in CYP1A1 were found based on age class or sex across all samples. However, within male false killer whales, juveniles expressed significantly higher levels of CYP1A1 when compared to adults. Total polychlorinated biphenyl (∑PCBs) concentrations in 84 % of false killer whales exceeded proposed threshold levels for health effects, and ∑PCBs correlated with CYP1A1 expression. There was no significant relationship between PCB toxic equivalent quotient and CYP1A1 expression, suggesting that this response may be influenced by agonists other than the dioxin-like PCBs measured in this study. No significant differences were found for CYP1A1 expression among social clusters of false killer whales. This work provides a foundation for future health monitoring of the endangered stock of false killer whales and other Hawaiian odontocetes.

It has been hypothesized for decades that environmental pollutants may contribute to green sea turtle fibropapillomatosis (FP), possibly through immunosuppression leading to greater susceptibility to the herpesvirus, the putative causative agent of this tumor-forming disease. To address this question, we measured concentrations of 164 persistent organic pollutants (POPs) and halogenated phenols in 53 Hawaiian green turtle (Chelonia mydas) plasma samples archived by the Biological and Environmental Monitoring and Archival of Sea Turtle Tissues (BEMAST) project at the National Institute of Standards and Technology Marine Environmental Specimen Bank. Four groups of turtles were examined: free-ranging turtles from Kiholo Bay (0% FP, Hawaii), Kailua Bay (low FP, 8%, Oahu), and Kapoho Bay (moderate FP, 38%, Hawaii) and severely tumored stranded turtles that required euthanasia (high FP, 100%, Main Hawaiian Islands). Four classes of POPs and seven halogenated phenols were detected in at least one of the turtles, and concentrations were low (often

Some cetaceans bioaccumulate substantial concentrations of planar halogenated aromatic hydrocarbons (PHAHs) in their tissues, but little is known about the effects of such burdens on cetacean health. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and related PHAHs cause toxicity via activation of the aryl hydrocarbon receptor (AHR), a member of the bHLH-PAS family of transcription factors. Differences in AHR structure and function are known to contribute to species-specific differences in susceptibility to PHAH toxicity. To ascertain the potential for PHAH effects in a cetacean, we characterized an AHR from the beluga whale, Delphinapterus leucas. The 3.2 kb cDNA encodes an 845-amino acid protein with a predicted size of 95.5 kDa. Overall, the beluga AHR shares 85% amino acid sequence identity with the human AHR and 75% identity with the mouse AHR Ah(b-1) allele. Beluga AHR protein synthesized in a rabbit reticulocyte lysate system demonstrated specific, high-affinity [(3)H]TCDD binding. Saturation binding analysis was used to compare the [(3)H]TCDD binding affinity of the in vitro-expressed beluga AHR with affinities of in vitro-expressed AHRs from a dioxin-sensitive mouse strain (Ah(b-1) allele) and humans. The beluga AHR bound [(3)H]TCDD with an affinity (K(d)= 0.43 +/- 0.16 nM) that was at least as high as that of the mouse AHR (K(d)= 0.68 +/- 0.23 nM), and significantly greater than that of the human AHR (K(d)= 1.63 +/- 0.64 nM). In electrophoretic mobility shift assays, the beluga AHR exhibited sequence-specific, Arnt-dependent binding to a dioxin responsive enhancer (DRE). Upon transient transfection into mammalian cells, the beluga AHR activated transcription of a luciferase reporter under control of a DRE-containing fragment of the mouse Cyp1a1 promoter. These results show that in an in vitro system, the beluga AHR possesses characteristics similar to those of AHRs from other mammals that are considered sensitive to toxic effects of PHAHs. Together, these results demonstrate that the use of in vitro-expressed proteins is a promising approach for addressing molecular and biochemical questions concerning PHAH toxicity in endangered or protected species.

To assess large-whale stocks following the cessation of land-based South Georgia whaling in 1965, we report three independent sighting databases: a cruise in 1997, observations from Bird Island (NW of South Georgia) between 1979 and 1998, and mariner sightings between 1992 and 1997. All species were rare, with sightings of southern right whales being the most common event. Two right whales photographed off South Georgia matched animals known from Peninsula Valdes, Argentina, a population known to be growing at 7%per annum. In contrast, blue and fin whales appeared to be less abundant. A single blue whale mother-calf pair was observed off the Shag Rocks in February 1997. Extirpation of animals from this particular feeding ground is the most likely reason for ongoing low numbers of all species. Other factors may include competition for krill by traditional predators such as penguins and seals and more recently by humans, an unusually high rate of natural mortality, habitat change such as alteration in sea ice coverage, and/or the impact of ongoing whaling. The history of this critical area of large-whale habitat and this report demonstrate the need for improved, consistent longterm monitoring of population trends for these depleted stocks.

Persistent organic pollutants (POPs) are toxic man-made chemicals that bioaccumulate and biomagnify in food webs, making them a ubiquitous threat to the marine environment. Although many studies have determined concentrations of POPs in top predators, no studies have quantified POPs in stranded cetaceans within the last 30 years around the Hawaiian Islands. A suite of POPs was measured in the blubber of 16 cetacean species that stranded in the tropical Pacific, including Hawai'i from 1997 to 2011. The sample set includes odontocetes (n=39) and mysticetes (n=3). Median (range) contaminant concentrations in ng/g lipid for the most representative species category (delphinids excluding killer whales [n=27]) are: 9650 (44.4-99,100) for ∑DDTs, 6240 (40.8-50,200) for ∑PCBs, 1380 (6.73-9520) for ∑chlordanes, 1230 (13.4-5510) for ∑toxaphenes, 269 (1.99-10,100) for ∑PBDEs, 280 (2.14-4190) for mirex, 176 (5.43-857) for HCB, 48.1 (5.42-566) for ∑HCHs, 33.9 (2.42-990) for ∑HBCDs, 1.65 (0.435-11.7) for octachlorostyrene and 1.49 (2.07-13.1) for pentachlorobenzene. ∑PCB concentrations in these Pacific Island cetaceans approach and sometimes exceed proposed toxic threshold values. Backward stepwise multiple regressions indicated the influence of life history parameters on contaminant concentrations when performed with three independent variables (species category, year of stranding, and sex/age class). No temporal trends were noted (p0.063), but sex/age class influences were evident with adult males exhibiting greater contaminant loads than adult females and juveniles for ∑DDT, ∑PCBs, ∑CHLs, and mirex (p≤0.036). POP concentrations were lower in mysticetes than odontocetes for many compound classes (p≤0.003). p,p'-DDE/∑DDTs ratios were greater than 0.6 for all species except humpback whales, suggesting exposure to an old DDT source. These POP levels are high enough to warrant concern and continued monitoring.

Laysan albatross (Phoebastria immutabilis) ingest plastic marine debris of a wide range of shape, sizes and sources. To better characterize this plastic and provide insights regarding its provenance and persistence in the environment, we developed a simple method to classify plastic fragments of unknown origin according to the resin codes used by the Society of Plastics Industry. Known plastics were analyzed by gas chromatography-mass spectroscopy (GC-MS) to identify indicator chemicals characteristic of each plastic resin. Application of this method to fragments of ingested plastic debris from boluses of Laysan albatross from Kure Atoll, Hawai'i, yielded proportions of 0.8% High Density Polyethylene, 6.8% Polysty- rene, 8.5% Polyethylene Terephthalate, 20.5% Polyvinyl Chloride and 68.4% Polypropylene. Some frag- ments were composed of multiple resin types. These results suggest that infrequently recycled plastics are the dominant fragments ingested by albatross, and that these are the most prevalent and persistent resin types in the marine environment.

We review the molecular and epidemiological characteristics of cetacean morbillivirus (CeMV) and the diagnosis and pathogenesis of associated disease, with six different strains detected in cetaceans worldwide. CeMV has caused epidemics with high mortality in odontocetes in Europe, the USAand Australia. It represents a distinct species within the Morbillivirusgenus. Although most CeMV strains are phylogenetically closely related, recent data indicate that morbilliviruses recovered from Indo-Pacific bottlenose dolphins (Tursiops aduncus), from Western Australia, and a Guiana dolphin (Sotalia guianensis), from Brazil, are divergent. The signaling lymphocyte activation molecule (SLAM) cell receptor for CeMV has been characterized in cetaceans. It shares higher amino acid identity with the ruminant SLAM than with the receptors of carnivores or humans, reflecting the evolutionary history of these mammalian taxa. In Delphinidae, three amino acid substitutions may result in a higher affinity for the virus. Infection is diagnosed by histology, immunohistochemistry, virus isolation, RT-PCR, and serology. Classical CeMV-associated lesions include bronchointerstitial pneumonia, encephalitis, syncytia, and lymphoid depletion associated with immunosuppression. Cetaceans that survive the acute disease may develop fatal secondary infections and chronic encephalitis. Endemicallyinfected, gregarious odontocetes probably serve as reservoirs and vectors. Transmission likely occurs through the inhalation of aerosolized virus but mother to fetus transmission was also reported. Publisher PDF

Mantle xenoliths hosted by the Historic Volcan de San Antonio, La Palma, Canary Islands, fall into two main group. Group I consists of spinel harzburgites, rare spinel lherzolites and spinel dunites, whereas group II comprises spinel wehrlites, amphibole wehrlites, and amphibole clinopyroxenites. We here present data on group I xenoliths, including veined harzburgites and dunites which provide an excellent basis for detailed studies of metasomatic processes. The spinel harzburgite and lherzolite xenoliths have modal ol−opx−cpx ratios and mineral and whole rock major element chemistry similar to those found in Lanzarote and Hierro, and are interpreted as highly refractory, old oceanic lithospheric mantle. Spinel dunites are interpreted as old oceanic peridotite which has been relatively enriched in olivine and clinopyroxene (and highly incompatible elements) through reactions with basaltic Canarian magmas, with relatively high melt/peridotite ratio. Group I xenoliths from La Palma differ from the Hierro and Lanzarote ones by a frequent presence of minor amounts of phlogopite (and amphibole). Metasomatic processes are also reflected in a marked enrichment of strongly incompatible relative to moderately incompatible trace elements, and in a tendency for Fe−Ti enrichment along grain boundaries in some samples. The veins in the veined xenoliths show a gradual change in phase assemblage and composition of each phase, from Fe−Ti-rich amphibole+augite+Fe−Ti-oxides+apatite+basaltic glass, to Ti-poor phlogopite+Cr-diopside±chromite+ Si−Na−K-rich glass+fluid. Complex reaction zones between veins and peridotite include formation of clinopyroxene±olivine+glass at the expense of orthopyroxene in harzburgite, and clinopyroxene+spinel±amphibole±glass at the expense of olivine in dunite. The dramatic change in glass composition from the broadest to the narrowest veins includes increasing SiO2 from 44 to 67 wt%, decreasing TiO2/Al2O3 ratio from >0.24 to about 0.02, and increasing K2O and Na2O from 1.8 to >7.0 wt% and 3.8 to 6.7 wt%, respectively. The petrographic observations supported by petrographic mixing calculations indicate that the most silicic melts in the veined xenoliths formed as the result of reaction between infiltrating basaltic melt and peridotite wall-rock. The highly silicic, alkaline melt may represent an important metasomatic agent. Pervasive metasomatism by highly silicic melts (and possibly fluids unmixed from these) may account for the enriched trace element patterns and frequent presence of phlogopite in the upper mantle under La Palma.

Some cetaceans bioaccumulate substantial concentrations of halogenated aromatic hydrocarbons (HAH) in their tissues, but little is known about the effects of such burdens on cetacean health. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and related HAH cause toxicity via activation of the aryl hydrocarbon receptor (AHR). Differences in AHR structure and function are known to contribute to species-specific differences in susceptibility to HAH toxicity. The goal of these studies was to ascertain the potential for HAH effects in cetaceans by characterizing the AHR from the beluga, Deiphinapterus leucas. The beluga AHR was characterized by its molecular structure, capacity for ligand binding, structure-binding relationships with various classes of HAH, as well as tissue-specific expression. These results show that: (1) in an in vitro system, the beluga AHR possesses binding affinities similar to AHRs of other mammals that are considered sensitive to toxic effects of HAH, (2) Structure-activity relationships are consistent with a common mechanism of coplanar HAH action among cetaceans and rodent species, and (3) the AHR protein is expressed in many tissues of the beluga, and is present at high levels in the lymphoid organs, liver and lung. Together, these data suggest that cetaceans can be considered sensitive to the action of coplanar HAH.

Background Bone marrow stromal cells produce cytokines required for the normal growth and development of all eight hematopoietic cell lineages. Aberrant cytokine production by stromal cells contributes to blood cell dyscrasias. Consequently, factors that alter stromal cell cytokine production may significantly compromise the development of normal blood cells. We have shown that environmental chemicals, such as aromatic hydrocarbon receptor (AhR) agonists, suppress B lymphopoiesis by modulating bone marrow stromal cell function. Here, we extend these studies to evaluate the potential for two prototypic AhR agonists, 7,12-dimethylbenz [a]anthracene (DMBA) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), to alter stromal cell cytokine responses. Methods Bone marrow stromal cells were treated with AhR agonists and bacterial lipopolysaccharide (LPS) to mimic innate inflammatory cytokine responses and to study the effects of AhR ligands on those responses. Steady state cytokine RNA levels were screened by RNAse protection assays (RPA) and quantified by real-time PCR. Cytokine (IL-6) protein production was measured by ELISA. NF-κB EMSAs were used to study IL-6 transcriptional regulation. Results RPAs indicated that AhR+ bone marrow stromal cells consistently up-regulated genes encoding IL-6 and LIF in response to LPS, presumably through activation of Toll-like receptor 4. Pre-treatment with low doses of DMBA or TCDD selectively abrogated IL-6 gene induction but had no effect on LIF mRNA. Real-time-PCR indicated a significant inhibition of IL-6 mRNA by AhR ligands within 1 hour of LPS challenge which was reflected in a profound down-regulation of IL-6 protein induction, with DMBA and TCDD suppressing IL-6 levels as much as 65% and 88%, respectively. This potent inhibitory effect persisted for at least 72 hours. EMSAs measuring NF-κB binding to IL-6 promoter sequences, an event known to induce IL-6 transcription, indicated a significant decrease in the LPS-mediated induction of DNA-binding RelA/p50 and c-Rel/p50 heterodimers in the presence of DMBA. Conclusions Common environmental AhR agonists can suppress the response to bacterial lipopolysaccharide, a model for innate inflammatory responses, through down-regulation of IL-6, a cytokine critical to the growth of several hematopoietic cell subsets, including early B cells. This suppression occurs at least at the level of IL-6 gene transcription and may be regulated by NF-κB.

The aryl hydrocarbon receptor (AHR) mediates the toxic and biochemical effects of 2,3,7,8-tetrachlorodibenzo- p -dioxin (TCDD) and related planar halogenated aromatic hydrocarbons (PHAH). Differences in AHR expression or function may contribute to species-specific and tissue-specific differences in susceptibility to PHAH. Odontocetes (toothed whales) bioaccumulate some of the highest PHAH tissue concentrations reported. To ascertain the potential for PHAH effect at the biochemical level, we cloned, sequenced, and expressed an AHR from beluga liver. The 3.2 kb cDNA encodes an 845 amino acid protein with a predicted size of 95.5 kDa. The beluga AHR expressed by in vitro transcription and translation exhibits specific, high-affinity binding of [ 3 H]-TCDD. We used several techniques to measure tissue-specific expression of the beluga AHR. Western blotting revealed a band of expected size in cytosols prepared from beluga liver, lung, thymus, and spleen, but not heart, kidney, or testis. Semi-quantitative analysis using RT-PCR detected expression of AHR transcripts in all tissues tested. Analysis of [ 3 H]-TCDD binding by velocity sedimentation on sucrose gradients detected specific binding in liver and lung cytosols, and a lesser amount in heart. cDNA cloning of susceptibility genes coupled with in vitro expression and functional analysis of the encoded proteins may serve as a valuable approach to characterize the susceptibility of protected species to PHAH-induced toxicity. (NIEHS Grant ES06272 and Sea Grant NA46RG0470 (R/B-137)).

Tveitite is a new mineral with formula of the type Ca 1−x ( Y, RE ) x F 2+x where x is approximately 0.3. It is found in a cleavelandite pegmatite at Hoydalen in Tordal, Telemark, S. Norway. The sub-cell is thought to be monoclinic, a ′=3.924, b ′=3.893, c ′=5.525, s=90°26′, Z =2 . The structure is very similar to that of fluorite and can be described in terms of a pseudo-cubic cell [1 1 0] 0 = 5.527, [110] 0 = 5.527, c = 5.525, α = 90°18′, γ = 89°37′ which contains approximately 1[YCa3F9]. The mineral shows twinning in at lest three different directions giving a grating structure probably due to high-low inversion. DTA studies show an inversion point at about 670°C. Material heated above this temperature gives X-ray diffraction patterns corresponding to yttrofluorite (a0 5.528). The effect of trivalent ions on the stability of fluorite is discussed.