Translocation of particles and inflammatory responses after exposure to fine particles and nanoparticles in an epithelial airway model

ABSTRACT: BACKGROUND: Experimental studies provide evidence that inhaled nanoparticles may translocate over the airspace epithelium and cause increased cellular inflammation. Little is known, however, about the dependence of particle size or material on translocation characteristics, inflammatory response and intracellular localization. RESULTS: Using a triple cell co-culture model of the human airway wall composed of epithelial cells, macrophages and dendritic cells we quantified the entering of fine (1 mum) and nano-sized (0.078 mum) polystyrene particles by laser scanning microscopy. The number distribution of particles within the cell types was significantly different between fine and nano-sized particles suggesting different translocation characteristics. Analysis of the intracellular localization of gold (0.025 mum) and titanium dioxide (0.02-0.03 mum) nanoparticles by energy filtering transmission electron microscopy showed differences in intracellular localization depending on particle composition. Titanium dioxide nanoparticles were detected as single particles without membranes as well as in membrane-bound agglomerations. Gold nanoparticles were found inside the cells as free particles only. The potential of the different particle types (different sizes and different materials) to induce a cellular response was determined by measurements of the tumour necrosis factor-alpha in the supernatants. We measured a 2-3 fold increase of tumour necrosis factor-alpha in the supernatants after applying 1 mum polystyrene particles, gold nanoparticles, but not with polystyrene and titanium dioxide nanoparticles. CONCLUSION: Quantitative laser scanning microscopy provided evidence that the translocation and entering characteristics of particles are size-dependent. Energy filtering transmission electron microscopy showed that the intracellular localization of nanoparticles depends on the particle material. Both particle size and material affect the cellular responses to particle exposure as measured by the generation of tumour necrosis factor-alpha.

Inhibition by interferon-gamma of human mononuclear cell-mediated low density lipoprotein oxidation. Participation of tryptophan metabolism along the kynurenine pathway

In this study we examined the potential inhibition by interferon-gamma (IFN gamma) of the early stages of low density lipoprotein (LDL) oxidation mediated by human peripheral blood mononuclear cells (PBMC) and monocyte-derived macrophages (MDM) in Ham's F-10 medium supplemented with physiological amounts of L-tryptophan (Trp). We assessed LDL oxidation by measuring the consumption of LDL's major antioxidant (i.e., alpha-tocopherol) and targets for oxidation (cholesteryllinoleate and cholesterylarachidonate), together with the accumulation of cholesterylester hydroperoxides and the increase in relative electrophoretic mobility of the lipoprotein particle. Exposure of PBMC or MDM to IFN gamma induced the degradation of extracellular Trp with concomitant accumulation of kynurenine, anthranilic and 3-hydroxyanthranilic acid (3HAA) in the culture medium. Formation of 3HAA, but neither Trp degradation nor formation of kynurenine and anthranilic acid, was inhibited by low amounts of diphenylene iodonium (DPI) in a concentration-dependent manner. In contrast to oxidative Trp metabolism, exposure of human PBMC or MDM to IFN gamma failed to induce degradation of arginine, and nitrite was not detected in the cell supernatant, indicating that nitric oxide synthase was not induced under these conditions. Incubation of LDL in Trp-supplemented F-10 medium resulted in a time-dependent oxidation of the lipoprotein that was accelerated in the presence of PBMC or MDM but inhibited strongly in the presence of both cells and IFN gamma, i.e., when Trp degradation and formation of 3HAA were induced. In contrast, when IFN gamma was added to PBMC or MDM in F-10 medium that was virtually devoid of Trp, inhibition of cell-accelerated LDL oxidation was not observed. Exogenous 3HAA added to PBMC or purified monocytes in the absence of IFN gamma also strongly and in a concentration-dependent manner inhibited LDL oxidation. Selective inhibition of IFN gamma-induced formation of 3HAA by DPI caused reversion of the inhibitory action of this cytokine on both PBMC- and MDM-mediated LDL oxidation. These results show that IFN gamma treatment of human PBMC or MDM in vitro attenuates the extent of LDL oxidation caused by these cells, and indicate that Trp degradation with formation of 3HAA is a major contributing factor to this inhibitory activity.

Direct combination of nanoparticle fabrication and exposure to lung cell cultures in a closed setup as a method to simulate accidental nanoparticle exposure of humans

The tremendous application potential of nanosized materials stays in sharp contrast to a growing number of critical reports of their potential toxicity. Applications of in vitro methods to assess nanoparticles are severely limited through difficulties in exposing cells of the respiratory tract directly to airborne engineered nanoparticles. We present a completely new approach to expose lung cells to particles generated in situ by flame spray synthesis. Cerium oxide nanoparticles from a single run were produced and simultaneously exposed to the surface of cultured lung cells inside a glovebox. Separately collected samples were used to measure hydrodynamic particle size distribution, shape, and agglomerate morphology. Cell viability was not impaired by the conditions of the glovebox exposure. The tightness of the lung cell monolayer, the mean total lamellar body volume, and the generation of oxidative DNA damage revealed a dose-dependent cellular response to the airborne engineered nanoparticles. The direct combination of production and exposure allows studying particle toxicity in a simple and reproducible way under environmental conditions.

Nitric oxide induces cell death in canine cruciate ligament cells by activation of tyrosine kinase and reactive oxygen species

BACKGROUND: There is increasing evidence suggesting that development of progressive canine cranial cruciate ligament (CCL) rupture involves a gradual degeneration of the CCL itself, initiated by a combination of factors, ranging from mechanical to biochemical. To date, knowledge is lacking to what extent cruciate disease results from abnormal biomechanics on a normal ligament or contrary how far preliminary alterations of the ligament due to biochemical factors provoke abnormal biomechanics. This study is focused on nitric oxide (NO), one of the potential biochemical factors. The NO-donor sodium nitroprusside (SNP) has been used to study NO-dependent cell death in canine cranial and caudal cruciate ligament cells and to characterize signaling mechanisms during NO-stimulation. RESULTS: Sodium nitroprusside increased apoptotic cell death dose- and time-dependently in cruciate ligamentocytes. Cells from the CCL were more susceptible to apoptosis than CaCL cells. Caspase-3 processing in response to SNP was not detected. Testing major upstream and signal transducing pathways, NO-induced cruciate ligament cell death seemed to be mediated on different levels. Specific inhibition of tyrosine kinase significantly decreased SNP-induced cell death. Mitogen activated protein kinase ERK1 and 2 are activated upon NO and provide anti-apoptotic signals whereas p38 kinase and protein kinase C are not involved. Moreover, data showed that the inhibition reactive oxygen species (ROS) significantly reduced the level of cruciate ligament cell death. CONCLUSIONS: Our data support the hypothesis that canine cruciate ligamentocytes, independently from their origin (CCL or CaCL) follow crucial signaling pathways involved in NO-induced cell death. However, the difference on susceptibility upon NO-mediated apoptosis seems to be dependent on other pathways than on these tested in the present study. In both, CCL and CaCL, the activation of the tyrosine kinase and the generation of ROS reveal important signaling pathways. In perspective, new efforts to prevent the development and progression of cruciate disease may include strategies aimed at reducing ROS.

Geochemical composition and granulometry of ODP Hole 167-1017E sediments

Intensification of North Pacific Intermediate Water during the Younger Dryas and stadials of the last glacial episode has been advocated by Kennett and his colleagues based on studies of ventilation history in Santa Barbara Basin. Because Santa Barbara Basin is a semi-isolated marginal basin, this hypothesis requires testing in sequences on the upper continental margin facing the open-ocean of the Pacific. Ocean Drilling Program Site 1017 is located on the upper slope of southern California off Point Conception close to the entrance of Santa Barbara Basin, an ideal location to test the hypothesis of late Quaternary switching in intermediate waters. We examined chemical and mineral composition, sedimentary structures, and grain size of hemipelagic sediments representing the last 80 k.y. at this site to detect changes in behavior of intermediate waters. We describe distinct compositional and textual variations that appear to reflect changes in grain size in response to flow velocity fluctuations of bottom waters. Qualitative estimates of changes in degree of pyritization indicate better ventilation of bottom water during intervals of stronger bottom-water flow. Comparison between variations in the sediment parameters and the planktonic d18O record indicates intensified bottom-current activity during the Younger Dryas and stadials of marine isotope Stage 3. This result strongly supports the hypothesis of Kennett and his colleagues. Our investigation also suggests strong grain-size control on organic carbon content (and to less extent carbonate carbon content). This, in turn, suggests the possibility that organic carbon content of sediments, which is commonly used as an indicator of surface productivity, can be influenced by bottom currents.

Clay mineraology and grain size composition of surface sediments from South Pacific Ocean

The reconstruction of low-latitude ocean-atmosphere interactions is one of the major issues of (paleo-)environmental studies. The trade winds, extending over 20° to 30° of latitude in both hemispheres, between the subtropical highs and the intertropical convergence zone, are major components of the atmospheric circulation and little is known about their long-term variability on geological time-scales, in particular in the Pacific sector. We present the modern spatial pattern of eolian-derived marine sediments in the eastern equatorial and subtropical Pacific (10°N to 25°S) as a reference data set for the interpretation of SE Pacific paleo-dust records. The terrigenous silt and clay fractions of 75 surface sediment samples have been investigated for their grain-size distribution and clay-mineral compositions, respectively, to identify their provenances and transport agents. Dust delivered to the southeast Pacific from the semi- to hyper-arid areas of Peru and Chile is rather fine-grained (4-8 µm) due to low-level transport within the southeast trade winds. Nevertheless, wind is the dominant transport agent and eolian material is the dominant terrigenous component west of the Peru-Chile Trench south of ~ 5°S. Grain-size distributions alone are insufficient to identify the eolian signal in marine sediments due to authigenic particle formation on the sub-oceanic ridges and abundant volcanic glass around the Galapagos Islands. Together with the clay-mineral compositions of the clay fraction, we have identified the dust lobe extending from the coasts of Peru and Chile onto Galapagos Rise as well as across the equator into the doldrums. Illite is a very useful parameter to identify source areas of dust in this smectite-dominated study area.

Mineralogy and major element oxide composition of sediments from ODP Hole 172-1063D

Dansgaard-Oeschger (D-O) cycles in sediment at Site 1063 are characterized by distinct fluctuations in physical properties. Stadials are marked by low bulk density and interstadials by high bulk density. Compressional (P-)wave velocity is in phase with bulk density over some but not all depth intervals. Four of the D-O cycles straddling the oxygen isotope Stage 4/5 boundary have been studied in detail to understand the origin of the physical properties changes. Sediment on the Bermuda Rise is comprised of three main components: calcite, aluminosilicate minerals, and biogenic silica. Calcite concentrations vary from 1% to 43% of bulk sediment and are highest during interstadials. Aluminosilicate concentrations vary from 52% to 92% of bulk sediment and are highest during stadials. The major element ratios Al2O3/TiO2 and K2O/Al2O3 show increases across bulk density cycles, suggesting a change in the composition of aluminosilicates. This interpretation is supported by mineralogical analyses, which show a subtle change in clay composition. Biogenic silica concentrations vary from 0% to 23% of bulk sediment and are also highest during stadials. However, the abundance of silica varies significantly from one D-O cycle to another. Silt and fine sand abundance also increase during the first of the four stadials. This coarsening of sediment coincides with the increase in biogenic silica. The low grain density and high porosity associated with biogenic silica result in intervals of low bulk-sediment density. The abundance of biogenic silica closely matches P-wave velocity, suggesting that silica imparts a greater rigidity to the sediment.

Geochemical provenance analysis of fine-grained sediment of sediment core AND1-1

Bulk chemical fine-grained sediment compositions from southern Victoria Land glacimarine sediments provide significant constraints on the reconstruction of sediment provenance models in the McMurdo Sound during Late Cenozoic time. High-resolution (~ 1 ka) geochemical data were obtained with a non-destructive AVAATECH XRF Core Scanner (XRF-CS) on the 1285 m long ANDRILL McMurdo Ice Shelf Project (MIS) sediment core AND-1B. This data set is complemented by high-precision chemical analyses (XRF and ICP-OES) on discrete samples. Statistical analyses reveal three geochemical facies which are interpreted to represent the following sources for the sediments recovered in the AND-1B core: 1) local McMurdo Volcanic Group (MVG) rocks, 2) Transantarctic Mountain rocks west of Ross Island (W TAM), and 3) Transantarctic Mountain rocks from more southerly areas (S TAM). Data indicate in combination with other sediment facies analyses (McKay et al., 2009, doi:10.1130/B26540.1) and provenance scenarios (Talarico and Sandroni, 2009, doi:10.1016/j.gloplacha.2009.04.007) that diamictites at the drill site are largely dominated by local sources (MVG) and are interpreted to indicate cold polar conditions with dry-based ice. MVG is interpreted to indicate cold polar condition with dry-based ice. A mixture of MVG and W TAM is interpreted to represent polar conditions and the S TAM facies is interpreted to represent open-marine conditions. Down-core variations in geochemical facies in the AND-1B core are interpreted to represent five major paleoclimate phases over the past 14 Ma. Cold polar conditions with major MVG influence occur below 1045 mbsf and above 120 mbsf. A section of warmer climate conditions with extensive peaks of S TAM influence characterizes the rest of the core, which is interrupted by a section from 525 to 855 mbsf of alternating influences of MVG and W TAM.

Grain size composition and age of ODP Hole 167-1017E sediments

Siliciclastic sedimentation at Ocean Drilling Program Site 1017 on the southern slope of the Santa Lucia Bank, central California margin, responded closely to oceanographic and climatic change over the past ~130 ka. Variation in mean grain-size and sediment sorting within the ~25-m-thick succession from Hole 1017E show Milankovitch-band to submillenial-scale variation. Mean grain size of the "sortable silt" fraction (10-63 µm) ranges from 17.6 to 33.9 µm (average 24.8 µm) and is inversely correlated with the degree of sorting. Much of the sediment has a bimodal or trimodal grain-size distribution that is composed of distinct fine silt, coarse silt to fine sand, and clay-size components. The position of the mode and the sorting of each component changes through the succession, but the primary variation is in the presence or abundance of the coarse silt fraction that controls the overall mean grain size and sorting of the sample. The occurrence of the best-sorted, finest grained sediment at high stands of sea level (Holocene, marine isotope Substages 5c and 5e) reflect the linkage between global climate and the sedimentary record at Site 1017 and suggest that the efficiency of off-shelf transport is a key control of sedimentation on the Santa Lucia Slope. It is not clear what proportion of the variation in grain size and sorting may also be caused by variations in bottom current strength and in situ hydrodynamic sorting.

Elemental analyses (XRF), total organic carbon (TOC), grain size analyses, lamination index of sediment core SO130-289KL

The Dansgaard-Oeschger oscillations and Heinrich events described in North Atlantic sediments and Greenland ice are expressed in the climate of the tropics, for example, as documented in Arabian Sea sediments. Given the strength of this teleconnection, we seek to reconstruct its range of environmental impacts. We present geochemical and sedimentological data from core SO130-289KL from the Indus submarine slope spanning the last ~ 80 kyr. Elemental and grain size analyses consistently indicate that interstadials are characterized by an increased contribution of fluvial suspension from the Indus River. In contrast, stadials are characterized by an increased contribution of aeolian dust from the Arabian Peninsula. Decadal-scale shifts at climate transitions, such as onsets of interstadials, were coeval with changes in productivity-related proxies. Heinrich events stand out as especially dry and dusty events, indicating a dramatically weakened Indian summer monsoon, potentially increased winter monsoon circulation, and increased aridity on the Arabian Peninsula. This finding is consistent with other paleoclimate evidence for continental aridity in the northern tropics during these events. Our results strengthen the evidence that circum-North Atlantic temperature variations translate to hydrological shifts in the tropics, with major impacts on regional environmental conditions such as rainfall, river discharge, aeolian dust transport, and ocean margin anoxia.