MODULATION OF HOST INTESTINAL MYOELECTRIC ACTIVITY BY LOCAL ANAPHYLAXIS: A COMPONENT OF PROTECTIVE IMMUNITY TO AN ENTERIC PARASITE (TRICHINELLA SPIRALIS, EIMERIA NIESCHULZI, BOWEL MOTILITY)

The hypothesis tested was that rapid rejection of Trichinella spiralis infective larvae from immunized rats following a challenge infection is associated with a local anaphylactic reaction, and this response should be reflected in altered small intestinal motility. The objective was to determine if altered gut smooth muscle function accompanies worm rejection based on the assumption that anaphylaxis in vivo could be detected by changes in intestinal smooth muscle contractile activity (ie. an equivalent of the Schultz-Dale reaction or in vitro anaphylaxis). The aims were to (1) characterize motility changes by monitoring intestinal myoelectric activity in conscious rats during the enteric phase of T. spiralis infection in immunized hosts, (2) detect the onset and magnitude of myoelectric changes caused by challenge infection in immunized rats, (3) determine the parasite stimulus causing changes, and (4) determine the specificity of host response to stimulation. Electrical slow wave frequency, spiking activity, normal interdigestive migrating myoelectric complexes and abnormal migrating action potential complexes were measured. Changes in myoelectric parameters induced by larvae inoculated into the duodenum of immune hosts differed from those associated with primary infection with respect to time of onset, magnitude and duration. Myoelectric changes elicited by live larvae could not be reproduced by inoculation of hosts with dead larvae, larval excretory-secretory products, or by challenge with a heterologous parasite, Eimeria nieschulzi. These results indicate that (1) local anaphylaxis is a component of the initial response to T. spiralis in immune hosts, since the rapid onset of altered smooth muscle function parallels in time the expression of rapid rejection of infective larvae, and (2) an active mucosal penetration attempt by the worm is necessary to elicit this host response. These findings provide evidence that worm rejection is a consequence of, or sequel to, an immediate hypersensitivity reaction elicited when parasites attempt to invade the gut mucosa of immunized hosts. ^

Cellular Uptake of Neutrohpil Elastase Links Inflammation to Adaptive Immunity

Many tumors arise from sites of inflammation providing evidence that innate immunity is a critical component in the development and progression of cancer. Neutrophils are primary mediators of the innate immune response. Upon activation, an important function of neutrophils is release of an assortment of proteins from their granules including the serine protease neutrophil elastase (NE). The effect of NE on cancer has been attributed primarily to its ability to degrade the extracellular matrix thereby promoting invasion and metastasis. Recently, it was shown that NE could be taken up by lung cancer cells leading to degradation of insulin receptor substrate-1 thereby promoting hyperactivity of the phosphatidylinositol-3 kinase (PI3K) pathway and tumor cell proliferation. To our knowledge, nobody has investigated uptake of NE by other tumor types. In addition, NE has broad substrate specificity suggesting that uptake of NE by tumor cells could impact processes regulating tumorigenensis other than activation of the PI3K pathway. Neutrophil elastase has been identified in breast cancer specimens where high levels of NE have prognostic significance. These studies have assessed NE levels in whole tumor lysates. Because the major source of NE is from activated neutrophils, we hypothesized that breast cancer cells do not have endogenous NE but may take up NE released by tumor associated neutrophils in the tumor microenvironment and that this could provide a link between the innate immune response to tumors and specific adaptive immune responses. In this thesis, we show that breast cancer cells lack endogenous NE expression and that they are able to take up NE resulting in increased generation of low molecular weight cyclin E (CCNE) and enhanced susceptibility to lysis by CCNE-specific cytotoxic T lymphocytes. We also show that after taking up NE and proteinase 3 (PR3), a second primary granule protease with significant homology to NE, breast cancer cells cross-present the NE- and PR3-derived peptide PR1 rendering them susceptible to PR1-targeted therapies. Taken together, our data support a role for NE uptake in modulating adaptive immune responses against breast cancer.

Human complement component C3 -binding proteins of Mycobacterium tuberculosis

Mycobacterium tuberculosis, the causative agent of tuberculosis, is a facultative intracellular pathogen that uses the host mononuclear phagocyte as a niche for survival and replication during infection. Complement component C3 has previously been shown to enhance the binding of M. tuberculosis to mononuclear phagocytes. Using a C3 ligand affinity blot protocol, we identified a 30 kDa C3-binding protein in M. tuberculosis as heparin-binding hemagglutinin (HbhA). HbhA was found to be a hydrophobic protein that localized to the cell membrane/cell wall fraction of M. tuberculosis, and this protein has previously been shown by others to be located on the surface of M. tuberculosis. The C3-binding activity of HbhA was localized to the C-terminus of the protein, which consists of lysine-alanine repeats. Full-length recombinant HbhA coated onto latex beads was shown to mediate the adherence of the beads to murine macrophage-like cells in both a C3-dependent and a C3-independent manner. An in-frame 576 by deletion in the hbhA gene was created in a virulent strain of M. tuberculosis using a PCR technique known as gene splicing by overlap extension (SOEing). Using the ΔhbhA mutant, HbhA was found not to be necessary for growth of M. tuberculosis in laboratory media or in macrophage-like cells, nor is HbhA required for adherence of M. tuberculosis to macrophage-like cells. HbhA is, however, required for infectivity of M. tuberculosis in mice. Mice infected with the ΔhbhA mutant show decreased growth in the lungs, liver, and spleen compared to mice infected with the wild-type strain. Using the ΔhbhA mutant strain, we were able to purify and identify a second 30-kDa C3-binding protein, HupB. These data demonstrate that HbhA is required for the in vivo but not the in vitro survival of M. tuberculosis and that HbhA is not necessary for the adherence of M. tuberculosis to the macrophage-like cells used in these studies. The expression of two proteins that bind human C3 may aid in the efficient binding of M. tuberculosis to complement receptors for uptake into mononuclear cells, or may influence other aspects of the host-parasite interaction. ^

Stable oxygen isotope record from a high-latitude reef off Western Australia

A core from a coral colony of Porites lutea was analysed for stable oxygen isotopic composition*. A 200-year proxy record of sea surface temperatures from the Houtman Abrolhos Islands off west Australia was obtained from coral delta18O. At 29°S, the Houtman Abrolhos are the southernmost major reef complex of the Indian Ocean. They are located on the path of the Leeuwin Current, a southward flow of warm, tropical water, which is coupled to Indonesian throughflow. Coral delta18O primarily reflects local oceanographic and climatic variability, which is largely determined by spatial variability of the Leeuwin Current. However, coherence between coral delta18O and the current strength itself is relatively weak. Evolutionary spectral and singular spectrum analyses of coral delta18O demonstrate a high variability in spectral composition through time. Oscillations in the 5-7-y, 14-15-y, and quasi-biennial bands reflect teleconnections of local sea surface temperature (SST) to tropical Pacific climate variability. Deviations between local (coral-based) and regional (instrument) SST contain a cyclic component with a period of 15 y. Coral delta18O suggests a rise in SST by 0.6°C since AD 1944, consistent with available instrumental SST records. A long-term warming by 1.4°C since AD 1795 is inferred from the coral record.

Water stable oxygen isotope records from six Antarctic ice core sites for the present and last interglacial periods

We compare the present and last interglacial periods as recorded in Antarctic water stable isotope records now available at various temporal resolutions from six East Antarctic ice cores: Vostok, Taylor Dome, EPICA Dome C (EDC), EPICA Dronning Maud Land (EDML), Dome Fuji and the recent TALDICE ice core from Talos Dome. We first review the different modern site characteristics in terms of ice flow, meteorological conditions, precipitation intermittency and moisture origin, as depicted by meteorological data, atmospheric reanalyses and Lagrangian moisture source diagnostics. These different factors can indeed alter the relationships between temperature and water stable isotopes. Using five records with sufficient resolution on the EDC3 age scale, common features are quantified through principal component analyses. Consistent with instrumental records and atmospheric model results, the ice core data depict rather coherent and homogenous patterns in East Antarctica during the last two interglacials. Across the East Antarctic plateau, regional differences, with respect to the common East Antarctic signal, appear to have similar patterns during the current and last interglacials. We identify two abrupt shifts in isotopic records during the glacial inception at TALDICE and EDML, likely caused by regional sea ice expansion. These regional differences are discussed in terms of moisture origin and in terms of past changes in local elevation histories, which are compared to ice sheet model results. Our results suggest that elevation changes may contribute significantly to inter-site differences. These elevation changes may be underestimated by current ice sheet models

Stable oxygen isotope record from Greenland ice cores

Twenty ice cores drilled in medium to high accumulation areas of the Greenland ice sheet have been used to extract seasonally resolved stable isotope records. Relationships between the seasonal stable isotope data and Greenland and Icelandic temperatures as well as atmospheric flow are investigated for the past 150-200 years. The winter season stable isotope data are found to be influenced by the North Atlantic Oscillation (NAO) and very closely related to SW Greenland temperatures. The linear correlation between the first principal component of the winter season stable isotope data and Greenland winter temperatures is 0.71 for seasonally resolved data and 0.83 for decadally filtered data. The summer season stable isotope data display higher correlations with Stykkisholmur summer temperatures and North Atlantic SST conditions than with SW Greenland temperatures. The linear correlation between Stykkisholmur summer temperatures and the first principal component of the summer season stable isotope data is 0.56, increasing to 0.66 for decadally filtered data. Winter season stable isotope data from ice core records that reach more than 1400 years back in time suggest that the warm period that began in the 1920s raised southern Greenland temperatures to the same level as those that prevailed during the warmest intervals of the Medieval Warm Period some 900-1300 years ago. This observation is supported by a southern Greenland ice core borehole temperature inversion. As Greenland borehole temperature inversions are found to correspond better with winter stable isotope data than with summer or annual average stable isotope data it is suggested that a strong local Greenland temperature signal can be extracted from the winter stable isotope data even on centennial to millennial time scales.

(Table 2) Mineralogy and carbon, oxygen, and hydrogen isotope ratios for secondary mineral samples from igneous rocks from ODP Hole 128-794D

The basaltic rocks of Hole 794D drilled during Leg 128 are strongly altered. Microprobe analyses and XRD spectra on small quantities of matter extracted from thin sections show that primary minerals and glassy zones of the groundmass are totally or partially replaced by clay minerals with chlorite/saponite mixed-layer composition whatever the rock sample considered. This mixed-layer was also identified in veins and vesicles where it crystallizes in spheroidal aggregates. The largest veins and vesicles are filled by a zoned deposit: the chlorite/saponite mixed-layer always occupies the central part and is rimmed by pure saponite. Calcite crystallizes in secondary fractures which crosscut the clayey veins and vesicles. Chemographic analysis based on the M+-4Si-3R2+ projection shows that the chemical composition of the saponite component in the mixed-layer is identical to that of the free saponite. This indicates that the clay mineral crystallization was controlled by the chemical composition of the alteration fluids. From petrographic evidence, it is suggested that both chlorite/saponite mixed-layer and free saponite belong to the same hydrothermal event and are produced by a temperature decrease. This is supported by the stable isotopic data. The isotopic data show very little variation: d18O saponite ranges from 13.1 per mil to 13.5 per mil, and dD saponite from -73.6 per mil to -70.0 per mil. d18O calcite varies from +19.7 per mil to +21.9 per mil vs SMOW and d13C from -3.2 per mil to +0.4 per mil vs. PDB. These values are consistent with seawater alteration of the basalt. The formation of saponite took place at 150°-180°C and the formation of calcite at about 65°C.

CaCO3, foraminifer fragmentation, benthic carbon isotopes, coarse lithic counts and radiogenic isotopes (Sr, Nd, Pb) of the Pliocene and earliest Pleistocene terrigenous component (3.3-2.4 Ma) of IODP Site 306-U1313

We present Plio-Pleistocene records of sediment color, %CaCO3, foraminifer fragmentation, benthic carbon isotopes (d13C) and radiogenic isotopes (Sr, Nd, Pb) of the terrigenous component from IODP Site U1313, a reoccupation of benchmark subtropical North Atlantic Ocean DSDP Site 607. We show that (inter)glacial cycles in sediment color and %CaCO3 pre-date major northern hemisphere glaciation and are unambiguously and consistently correlated to benthic oxygen isotopes back to 3.3 million years ago (Ma) and intermittently so probably back to the Miocene/Pliocene boundary. We show these lithological cycles to be driven by enhanced glacial fluxes of terrigenous material (eolian dust), not carbonate dissolution (the classic interpretation). Our radiogenic isotope data indicate a North American source for this dust (~3.3-2.4 Ma) in keeping with the interpreted source of terrestrial plant wax-derived biomarkers deposited at Site U1313. Yet our data indicate a mid latitude provenance regardless of (inter)glacial state, a finding that is inconsistent with the biomarker-inferred importance of glaciogenic mechanisms of dust production and transport. Moreover, we find that the relation between the biomarker and lithogenic components of dust accumulation is distinctly non-linear. Both records show a jump in glacial rates of accumulation from Marine Isotope Stage, MIS, G6 (2.72 Ma) onwards but the amplitude of this signal is about 3-8 times greater for biomarkers than for dust and particularly extreme during MIS 100 (2.52 Ma). We conclude that North America shifted abruptly to a distinctly more arid glacial regime from MIS G6, but major shifts in glacial North American vegetation biomes and regional wind fields (exacerbated by the growth of a large Laurentide Ice Sheet during MIS 100) likely explain amplification of this signal in the biomarker records. Our findings are consistent with wetter-than-modern reconstructions of North American continental climate under the warm high CO2 conditions of the Early Pliocene but contrast with most model predictions for the response of the hydrological cycle to anthropogenic warming over the coming 50 years (poleward expansion of the subtropical dry zones).

Stable carbon and oxygen isotope compositions of benthic foraminifera N. truempyi samples and coarse fraction analyses from ODP Site 198-1209

We have compiled the first stratigraphically continuous high-resolution benthic foraminiferal stable isotope record for the Paleocene from a single site utilizing cores recovered at Pacific ODP Site 1209. The long-term trend in the benthic isotope record suggests a close coupling of volcanic CO2 input and deep sea warming. Over the short-term the record is characterized by slow excursions with a pronounced periodic beat related to the short (100-kyr) and long (405-kyr) eccentricity cycle. The phase relationship between the benthic isotope record and eccentricity is similar to patterns documented for the Oligocene and Miocene confirming the role of orbital forcing as the pace maker for paleoclimatic variability on Milankovitch time scales. In addition, the record documents an unusual transient warming of 2°C coeval with a 0.6 per mil carbon isotope excursion and a decrease in carbonate content at 61.75 Ma. This event, which bears some resemblance to Eocene hyperthermals, marks the onset of a long-term decline in d13C. The timing indicates it might be related to the initiation of volcanism along Greenland margin.

Benthic oxygen flixes on the Washington shelf and slope

Benthic oxygen fluxes calculated from in situ microelectrode profiles arc compared with benthic flux chamber O2 uptake measurements on a transect of eight stations across the continental shelf and three stations on the slope of Washington State. Station depths ranged from 40 to 630 m and bottom-water oxygen concentrations were 127-38 µM. The fluxes measured by the two methods were similar on the slope, but on the shelf, the chamber flux exceeded the microelectrode flux by as much as a factor of 3-4. We attribute this difference to pore-water irrigation, a process which apparently accounts for the oxidation of a significant amount of organic C in the continental shelf sediments. Combining our diffusive flux data with other data demonstrates clearly that the bottomwater oxygen concentration must play some significant role in determining the sedimentary oxygen consumption rate. Numerical simulation of the microelectrode 0, profiles suggests that roughly half the diffusive 0, flux must be consumed within - 1 mm of the sediment surface. If this conclusion is correct, then the magnitude of the diffusive flux depends both on the bottom-water oxygen concentration and on the supply rate of labile C to the sediment surf'ace.

Mineralogy and stable oxygen isotope ratios of the Cretaceous-Tertiary boundary

Results of detailed mineralogical, chemical, and oxygen isotope analyses of the clay minerals and zeolites from two Cretaceous-Tertiary (K/T) boundary regions, Stevns Klint, Denmark, and Deep Sea Drilling Project (DSDP) Hole 465A in the north central Pacific Ocean, are presented. In the central part of the Stevns Klint K/T boundary layer, the only clay mineral detected by x-ray diffraction is a pure smectite with > 95 percent expandable layers. No detrital clay minerals or quartz were observed in the clay size fraction in these beds, whereas the clay minerals above and below the boundary layer are illite and mixed-layer smectite-illite of detrital origin as well as quartz. The mineralogical purity of the clay fraction, the presence of smectite only at the boundary, and the d18O value of the smectite (27.2 ± 0.2 per mil) suggest that it formed in situ by alteration of glass. Formation from impact rather than from volcanic glass is supported by its major element chemistry. The high content of iridium and other siderophile elements is not due to the cessation of calcium carbonate deposition and resulting slow sedimentation rates. At DSDP Hole 465A, the principal clay mineral in the boundary zone (80 to 143 centimeters) is a mixed-layer smectite-illite with >=90 percent expandable layers, accompanied by some detrital quartz and small amounts of a euhedral authigenic zeolite (clinoptilolite). The mixed-layer smectite-illite from the interval 118 to 120 centimeters in the zone of high iridium abundance has a very low rare earth element content; the negative cerium anomaly indicates formation in the marine environment. This conclusion is corroborated by the d18O value of this clay mineral (27.1 ± 0.2 per mil). Thus, this mixed-layer smectite-illite formed possibly from the same glass as the K/T boundary smectite at Stevns Klint, Denmark.

Planktic foraminiferal Mg/Ca and oxygen isotopes, plus estimated d18Oseawater and ice volume corrected d18Oseawater for the past 35000 years of sediment core GeoB10069-3

Evidence from geologic archives suggests that there were large changes in the tropical hydrologic cycle associated with the two prominent northern hemisphere deglacial cooling events, Heinrich Stadial 1 (HS1; ~19 to 15 kyr BP; kyr BP = 1000 yr before present) and the Younger Dryas (~12.9 to 11.7 kyr BP). These hydrologic shifts have been alternatively attributed to high and low latitude origin. Here, we present a new record of hydrologic variability based on planktic foraminifera-derived d18O of seawater (d18Osw) estimates from a sediment core from the tropical Eastern Indian Ocean, and using 12 additional d18Osw records, construct a single record of the dominant mode of tropical Eastern Equatorial Pacific and Indo-Pacific Warm Pool (IPWP) hydrologic variability. We show that deglacial hydrologic shifts parallel variations in the reconstructed interhemispheric temperature gradient, suggesting a strong response to variations in the Atlantic Meridional Overturning Circulation and the attendant heat redistribution. A transient model simulation of the last deglaciation suggests that hydrologic changes, including a southward shift in the Intertropical Convergence Zone (ITCZ) which likely occurred during these northern hemisphere cold events, coupled with oceanic advection and mixing, resulted in increased salinity in the Indonesian region of the IPWP and the eastern tropical Pacific, which is recorded by the d18Osw proxy. Based on our observations and modeling results we suggest the interhemispheric temperature gradient directly controls the tropical hydrologic cycle on these time scales, which in turn mediates poleward atmospheric heat transport.

Paleomagnetic, stable oxygen isotope ratios, and U/Th activities of ODP Site 172-1062

Geomagnetic excursions are recognized as intrinsic features of the Earth's magnetic field. High-resolution records of field behaviour, captured in marine sedimentary cores, present an opportunity to determine the temporal and geometric character of the field during geomagnetic excursions and provide constraints on the mechanisms producing field variability. We present here the highest resolution record yet published of the Blake geomagnetic excursion (~125 ka) measured in three cores from Ocean Drilling Program (ODP) Site 1062 on the Blake-Bahama Outer Ridge. The Blake excursion has a controversial structure and timing but these cores have a sufficiently high sedimentation rate (~10cm/ka) to allow detailed reconstruction of the field behaviour at this site during the excursion. Palaeomagnetic measurements of the cores reveal rapid transitions (<500 yr) between the contemporary stable normal polarity and a completely reversed state of long duration which spans a stratigraphic interval of 0.7 m. We determine the duration of the reversed state during the Blake excursion using oxygen isotope stratigraphy, combined with 230Th excess measurements to assess variations in the sedimentation rates through the sections of interest. This provides an age and duration for the Blake excursion with greater accuracy and with constrained uncertainty. We date the directional excursion as falling between 129 and 122 ka with a duration for the deviation of 6.5±1.3 kyr. The long duration of this interval and the fully reversed field suggest the existence of a pseudo-stable, reversed dipole field component during the excursion and challenge the idea that excursions are always of short duration.

Strontium and Oxygen isotope compositions from DSDP Holes 54-424A, 54-424B and 70-509B

Strontium and O isotope compositions of green clay minerals from sediment cores of three boreholes drilled into (sites 424A and 509B) and close to a hydrothermal mound (site 424B) near the Galapagos Spreading Center (DSDP Legs 54 and 70) were determined. The green clays consist mostly of a transition from Fesmectite (nontronite) to glauconite. 87Sr/86Sr ratios were measured on clay size-fractions after gentle acid leaching and on the recovered leachates from different samples. The 87Sr/86Sr ratios of the clay residues from both the 424A and B sites are well below the modern seawater value, which points consistently to precipitation from hydrothermal fluids that contained variable amounts of seawater, even away from mound. However, most of the clay residues from mound site 509B have 87Sr/86Sr ratios significantly above the seawater value, suggesting the occurrence of a detrital component together with the new authigenic particles. The clay minerals of the hydrothermal mound are mixed with detrital components, and that of the sample taken outside but near the mound as a reference for the surrounding oceanic environment, yields a hydrothermal signature. Crystallization temperatures of the clays range from 32 to 63 °C assuming a d18O value of +2.2 per mil for the mineralizing fluids. Hydrothermal fluids generated in the underlying oceanic crust, mixed in varied proportions with ambient seawater and migrated into beds of the mound in a sequence of recurrent processes that ultimately resulted in the formation of the observed clay minerals. No significant temperature differences were detected for crystallization of the K-rich glauconite and K-depleted nontronite. The 87Sr/86Sr ratios of the Sr leached off the clay particles are near the value of modern seawater, inferring a progressive replacement of the hydrothermal fluids by seawater in the pore space of the mound sediments.

Mineral and oxygen isotopic compositions in a profile through the altered lower oceanic crust, ODP Hole 176-735B

ODP Hole 735B located on the Southwest Indian Ridge at 57°E is an in situ sampled long, continuous section of lower oceanic crust. Oxygen isotope compositions of constituent minerals of Leg 176 gabbros have been measured by UV-laser oxygen isotope microprobe. Together with existing data from Leg 118, a complete oxygen isotope profile through the lower oceanic crust has been obtained. Most clinopyroxenes and olivines have normal mantle values of ~5.5 per mil and ~5.2 per mil, respectively, while plagioclases show slight d18O enrichment relative to its mantle value of 6.1per mil. Down-hole variations of Hole 735B gabbro indicate a downward decreasing d18O profile, with a kink at a depth of about 800 m below sea floor. Above this depth, gabbros are depleted in 18O relative to unaltered basalts, while below ~800 m they show nearly unmodified d18O values. Abundant seawater penetration appears to be limited to the upper part of the lower crust at ODP site 735 (~800 m into the gabbroic layer and ~2-2.5 km into the oceanic crust from the top of pillow basalts). Mass balance calculations show that the lower crust formed under this ultra-slow-spreading ridge has an average d18O value of 5.5 per mil. The whole crust at Site 735 has an overall 18O enrichment with d18O values of 6.0 per mil to 7.8 per mil, depending on the possible variation of the d18O values of the upper pillow basalts and sheeted dykes. The apparent difference in oxygen isotope compositions of ocean crusts formed with different spreading rates has important implications on the buffering of ocean water over geological time, as well as on the oxygen recycling between crust and mantle through subduction. The difference of seawater penetration between fast- and slow-spreading ridges could be related to their particular magmatic-tectonic history during the formation and aging of the crust. However, more analyses on continuous sections through oceanic and ophiolitic crust in different tectonic settings are required to derive any predictive models.