Particulate Ligands Research Articles

Electroanalytical Trace Metal Cations Quantification and Speciation in Freshwaters: Historical Overview, Critical Review of the Last Five Years and Road Map for Developing Dynamic Speciation Field Measurements.

An historical overview covering the field of electroanalytical metal cations speciation in freshwaters is presented here, detailing both the notable experimental and theoretical developments. Then, a critical review of the progress in the last five years is given, underlining in particular the improvements in electrochemical setups and methodologies dedicated to field surveys. Given these recent achievements, a road map to carry out on-site dynamic metal speciation measurements is then proposed, and the key future developments are discussed. This review shows that electroanalytical stripping techniques provide a unique framework for quantitatively assessing metals at trace levels while offering access to both thermodynamic and dynamic features of metal complexation with natural colloidal and particulate ligands.

Iron organic speciation during the LOHAFEX experiment: Iron ligands release under biomass control by copepod grazing

Abstract The LOHAFEX iron fertilization experiment consisted in the fertilization of the closed core of a cyclonic eddy located south of the Antarctic Polar Front in the Atlantic sector of the Southern Ocean. This eddy was characterized by high nitrate and low silicate concentrations. Despite a 2.5 fold increase of the chlorophyll-a (Chl-a) concentrations, the composition of the biological community did not change. Phytoplankton biomass was mostly formed by small autotrophic flagellates whereas zooplankton biomass was mostly comprised by the large copepod Calanus simillimus. Efficient recycling of copepod fecal pellets (the main component of the downward flux of organic matter) in the upper 100–150 m of the water column prevented any significant deep export of particulate organic carbon (POC). Before fertilization, dissolved iron (DFe) concentrations in the upper 200 m were low, but not depleted, at ~0.2 nM. High DFe concentrations appeared scattered from day 14 onwards as a result of the grazing activity. A second fertilization on day 21 had no significant effect on the DFe and Chl-a standing stocks. Work with unfiltered samples using different acidification protocols revealed that, by midway of LOHAFEX, rapid recycling of iron-replenished copepod fecal pellets explained the source of bioavailable iron that prolonged the duration of the bloom for many weeks. Here we present the evolution of the organic speciation of iron in the upper 200 m of the water column during LOHAFEX by a Competing Ligand Equilibrium method using voltammetry. During the first 12 days of the experiment, ligands of an affinity for iron similar to the ligands found before fertilization (logK′Fe′L~11.9) accumulated in fertilized waters mostly in the upper 80 m (from ~1 nM to ~2.5 nM). The restriction of ligand accumulation to the depth of Chl-a penetration points to exudation by the growing autotrophic population as the initial source of ligands. From day 5 onwards, we found in many samples a new class of ligands (L1) characterized by a significant higher conditional stability constant than the background complexation (logK′Fe′L1~12.9). During the middle section of the experiment (days 12 to 25) the accumulation of overall ligands and specifically L1, reached an upper limit in surface waters (at ~3 nM). Overall ligands and L1 accumulation was also observed below the mixed layer depth indicating that grazing was the process behind ligand release. During the last 10 days of the experiment ligands kept accumulating in deep waters but suffered a small decrease in the upper 50 m of the water column caused by the vanishing of L1. Ligand removal restricted to the euphotic layer was probably caused by photodegradation. A high correlation between [DFe] and [L1] suggested that recycled iron (released during grazing and copepod fecal pellet cycling) was in the form of FeL1 complexes. We hypothesize that the iron binding ligands released to the dissolved phase during LOHAFEX were mostly photosensitive intracellular ligands rapidly degraded in extracellular conditions (e.g.: pigments). Sloppy feeding by copepods and recycling of cells and cellular material in copepod fecal pellets caused the transfer of particulate ligands to the dissolved phase as zooplankton built up as a response to the blooming community.

Chemodynamics of metal ion complexation by charged nanoparticles: a dimensionless rationale for soft, core-shell and hard particle types.

Soft nanoparticulate complexants are defined by a spatial confinement of reactive sites and electric charges inside their 3D body. In turn, their reactivity with metal ions differs significantly from that of simple molecular ligands. A revisited form of the Eigen mechanism recently elucidated the processes leading to metal/soft particle pair formation. Depending on e.g. particle size and metal ion nature, chemodynamics of nanoparticulate metal complexes is controlled by metal conductive diffusion to/from the particles, by intraparticulate complex formation/dissociation kinetics, or by both. In this study, a formalism is elaborated to achieve a comprehensive and systematic identification of the rate-limiting step governing the overall formation and dissociation of nanoparticulate metal complexes. The theory covers the different types of spherical particulate complexants, i.e. 3D soft/permeable and core-shell particles, and hard particles with reactive sites at the surface. The nature of the rate-limiting step is formulated by a dynamical criterion involving a power law function of the ratio between particle radius and an intraparticulate reaction layer thickness defined by the key electrostatic, diffusional and kinetic components of metal complex formation/dissociation. The analysis clarifies the intertwined contributions of particle properties (size, soft or hard type, charge, density or number of reactive sites) and aqueous metal ion dehydration kinetics in defining the chemodynamic behavior of nanoparticulate metal complexes. For that purpose, fully parameterized chemodynamic portraits involving the defining features of particulate ligand and metal ion as well as the physicochemical conditions in the local intraparticulate environment, are constructed and thoroughly discussed under conditions of practical interest.

Novel nanocomposite materials for efficient and selective mercury ions capturing from wastewater

This work explored the particulate ligand impregnated highly ordered mesoporous silica based optical nanocomposite material for the sensitive and selective detection and removal of mercury (Hg(II)) from polluted water solutions. The ligand of N,N-disalicylidene-4,5-dimethyl-phenylenedene was prepared and successfully anchored onto the highly ordered mesoporous silica by the indirect approach in the preparation of nanocomposite materials. The materials were produced an optical color signal that was easily generated and transduced even at trace levels of Hg(II) ions. Considering the environmental factors, the nanocomposite materials were solvent-free systems for sensitive Hg(II) ions detection system. Adding of Hg(II) ions, the nanocomposite materials generated absorption peak and enhance the selective chromogenic behavior toward Hg(II) ions from pale yellow to dark read, which could be easily visualized by the naked eye. The ability to remove Hg(II) ions by the materials was performed relatively to different experimental parameters such as solution pH, reaction time, initial metal ions concentration and different competing metal ions. The Hg(II) ions adsorption reached an equilibrium within short time and the maximum monolayer adsorption capacity was 179.74mg/g. Also the foreign ions did not any significant affect for selective Hg(II) ions removal from water solutions under the optimum conditions. The elution was performed and the regeneration was possible during the elution operation. In addition, the nanocomposite materials could to reuse in several cycles without loss of structural ordering, permitting the reusability for further selective detection and separation of Hg(II) ions from water. In terms of the sensitivity and selectivity, the nanocomposite material is a promising candidate employed for in situ environmental pollution incidents, especially competent for the detection and removal of Hg(II) ions with an extremely high efficiency.

PI(3)P-p40phox binding regulates NADPH oxidase activation in mouse macrophages and magnitude of inflammatory responses in vivo.

Mutations in the leukocyte NADPH oxidase that abrogate superoxide production result in chronic granulomatous disease (CGD), an inherited immunodeficiency associated with recurrent infections and inflammatory complications. The cytosolic regulatory subunit p40phox plays a specialized role in stimulating NADPH oxidase activity on intracellular membranes via its phosphatidylinositol 3-phosphate [PI(3)P]-binding domain, as revealed by studies largely focused on neutrophils. Whether PI(3)P-p40phox-regulated superoxide production contributes to regulating inflammatory responses is not well understood. Here, we report that mice expressing p40phox R58A, which lacks PI(3)P binding, had impaired macrophage NADPH oxidase activity and increased sterile inflammation. p40phoxR58A/R58A macrophages exhibited diminished phagosome reactive oxygen species (ROS) in response to certain particulate and soluble ligands, including IgG-opsonized particles and a TLR2 agonist, along with unexpected defects in plasma membrane oxidase activity. Compared with wild-type (WT) mice, p40phoxR58A/R58A mice had elevated numbers of newly recruited neutrophils and monocytes in peritoneal inflammation elicited by zymosan, monosodium urate (MSU) crystals, or sodium periodate. At later time points, higher numbers of inflammatory macrophages in p40phoxR58A/R58A mice were consistent with delayed resolution. Our studies demonstrate a critical role of PI(3)P-p40phox binding for optimal activation of the NADPH oxidase in macrophages. Furthermore, selective loss of PI(3)P-regulated NADPH oxidase activity was sufficient to enhance significantly responses to inflammation and delay resolution.

CR3 and Dectin-1 Collaborate in Macrophage Cytokine Response through Association on Lipid Rafts and Activation of Syk-JNK-AP-1 Pathway.

Collaboration between heterogeneous pattern recognition receptors (PRRs) leading to synergistic coordination of immune response is important for the host to fight against invading pathogens. Although complement receptor 3 (CR3) and Dectin-1 are major PRRs to detect fungi, crosstalk between these two receptors in antifungal immunity is largely undefined. Here we took advantage of Histoplasma capsulatum which is known to interact with both CR3 and Dectin-1 and specific particulate ligands to study the collaboration of CR3 and Dectin-1 in macrophage cytokine response. By employing Micro-Western Array (MWA), genetic approach, and pharmacological inhibitors, we demonstrated that CR3 and Dectin-1 act collaboratively to trigger macrophage TNF and IL-6 response through signaling integration at Syk kinase, allowing subsequent enhanced activation of Syk-JNK-AP-1 pathway. Upon engagement, CR3 and Dectin-1 colocalize and form clusters on lipid raft microdomains which serve as a platform facilitating their cooperation in signaling activation and cytokine production. Furthermore, in vivo studies showed that CR3 and Dectin-1 cooperatively participate in host defense against disseminated histoplasmosis and instruct adaptive immune response. Taken together, our findings define the mechanism of receptor crosstalk between CR3 and Dectin-1 and demonstrate the importance of their collaboration in host defense against fungal infection.

FADD and caspase-8 mediate priming and activation of the canonical and non-canonical Nlrp3 inflammasomes (INC5P.321)

Abstract The Nlrp3 inflammasomes are essential for protection against several infections and autoimmune diseases. Despite the plethora of research focused on understanding the roles of NLRP3 in various disease settings, the mechanisms controlling its activation remain enigmatic. Herein, we show through genetic studies that FADD or caspase-8 is required for both transcriptional priming and posttranslational activation of the canonical and non-canonical Nlrp3 inflammasomes. FADD and caspase-8 were required for caspase-1 activation during canonical Nlrp3 inflammasome activation by both soluble (LPS+ATP) and particulate (Silica) ligands. We showed that FADD and caspase-8 also regulate Nlrp3 expression. Furthermore, both FADD and caspase-8 were critical for caspase-11 and caspase-1 activation during non-canonical Nlrp3 inflammasome activation by C. rodentium and E. coli. These requirements of FADD and caspase-8 for caspase-1 activation were specific to the Nlrp3 inflammasome since the activation of Nlrc4 inflammasome by S. typhimurium was not hampered by deletion of FADD or caspase-8. In vivo LPS challenge or C. rodentium infection of FADD or caspase-8-deficient mice resulted in impaired IL-1β production compared to their littermate controls. Thus, our results demonstrate that FADD and caspase-8 are critical mediators of the canonical and non-canonical Nlrp3 inflammasomes.

CD36 coordinates NLRP3 inflammasome activation by facilitating intracellular nucleation of soluble ligands into particulate ligands in sterile inflammation

Particulate ligands including cholesterol crystals and amyloid fibrils induce NLRP3-dependent production of interleukin-1β (IL-1β) in atherosclerosis, Alzheimer's disease and diabetes. Soluble endogenous ligands including oxidized-LDL, amyloid-β and amylin peptides accumulate in these diseases. Here we identify a CD36-mediated endocytic pathway that coordinates the intracellular conversion of these soluble ligands to crystals or fibrils, resulting in lysosomal disruption and NLRP3-inflammasome activation. Consequently, macrophages lacking CD36 failed to elicit IL-1β production in response to these ligands and targeting CD36 in atherosclerotic mice reduced serum IL-1β and plaque cholesterol crystal accumulation. Collectively, these findings highlight the importance of CD36 in the accrual and nucleation of NLRP3 ligands from within the macrophage and position CD36 as a central regulator of inflammasome activation in sterile inflammation.

Coupled particulate and continuum model for nanoparticle targeted delivery

Prediction of nanoparticle (NP) distribution in a vasculature involves transport phenomena at various scales and is crucial for the evaluation of NP delivery efficiency. A combined particulate and continuum model is developed to model NP transport and delivery processes. In the particulate model ligand–receptor binding kinetics is coupled with Brownian dynamics to study NP binding on a microscale. An analytical formula is derived to link molecular level binding parameters to particulate level adhesion and detachment rates. The obtained NP adhesion rates are then coupled with a convection–diffusion–reaction model to study NP transport and delivery at macroscale. The binding results of the continuum model agree well with those from the particulate model. The effects of shear rate, particle size and vascular geometry on NP adhesion are investigated. Attachment rates predicted by the analytical formula also agree reasonably well with the experimental data reported in literature. The developed coupled model that links ligand–receptor binding dynamics to NP adhesion rate along with macroscale transport and delivery processes may serve as a faster evaluation and prediction tool to determine NP distribution in complex vascular networks.

Spatial Distribution of the Particulate Organic Ligand in the South Pacific Ocean Surface Waters

Strong organic ligand in oceanic Particulate Matter (PM), as determined from amount of thorium adsorption onto PM in 0.1 M HCl, was measured for surface waters of mid-latitude region (ca. 32.5°S) in the South Pacific Ocean. The Particulate Organic Ligand (POL) concentrations in surface waters of the South Pacific, ranging from 2.6 to 13.3 nM, showed a longitudinal variation with low concentrations in the South Pacific subtropical gyre (180°-100°W) and high in the eastern South Pacific. The POL concentrations in the South Pacific surface waters increased with increasing nutrients (nitrate and phosphate) and chlorophyll concentration, which means that POL in surface waters is closely related to biological activity in the ocean; the high POL occurs in highly productivity region, whereas the low POL appears in the oligotrophic ocean. This is a first basin-scale measurement of POL in surface waters of the South Pacific. POL is a useful tool for better understanding of biological activity, particle dynamics and trace metal cycling in surface layer, if we have more detail knowledge on chemical and biogeochemical properties of POL, as do factors controlling spatial and temporal variability.

Adaptor Protein-3 in Dendritic Cells Facilitates Phagosomal Toll-like Receptor Signaling and Antigen Presentation to CD4+ T Cells

Effective major histocompatibility complex-II (MHC-II) antigen presentation from phagocytosed particles requires phagosome-intrinsic Toll-like receptor (TLR) signaling, but the molecular mechanisms underlying TLR delivery to phagosomes and how signaling regulates antigen presentation are incompletely understood. We show a requirement in dendritic cells (DCs) for adaptor protein-3 (AP-3) in efficient TLRrecruitment to phagosomes and MHC-II presentation of antigens internalized by phagocytosis butnot receptor-mediated endocytosis. DCs from AP-3-deficient pearl mice elicited impaired CD4(+) Tcell activation and Th1 effector cell function to particulate antigen invitro and to recombinant Listeria monocytogenes infection invivo. Whereas phagolysosome maturation and peptide:MHC-II complex assembly proceeded normally in pearl DCs, peptide:MHC-II export to the cell surface was impeded. This correlated with reduced TLR4 recruitment and proinflammatory signaling from phagosomes by particulate TLR ligands. We propose that AP-3-dependent TLR delivery from endosomes to phagosomes and subsequent signaling mobilize peptide:MHC-II export from intracellular stores.

Identification of Drosophila Yin and PEPT2 as Evolutionarily Conserved Phagosome-associated Muramyl Dipeptide Transporters

NOD2 (nucleotide-binding oligomerization domain containing 2) is an important cytosolic pattern recognition receptor that activates NF-kappaB and other immune effector pathways such as autophagy and antigen presentation. Despite its intracellular localization, NOD2 participates in sensing of extracellular microbes such as Staphylococcus aureus. NOD2 ligands similar to the minimal synthetic ligand muramyl dipeptide (MDP) are generated by internalization and processing of bacteria in hydrolytic phagolysosomes. However, how these derived ligands exit this organelle and access the cytosol to activate NOD2 is poorly understood. Here, we address how phagosome-derived NOD2 ligands access the cytosol in human phagocytes. Drawing on data from Drosophila phagosomes, we identify an evolutionarily conserved role of SLC15A transporters, Drosophila Yin and PEPT2, as MDP transporters in fly and human phagocytes, respectively. We show that PEPT2 is highly expressed by human myeloid cells. Ectopic expression of both Yin and PEPT2 increases the sensitivity of NOD2-dependent NF-kappaB activation. Additionally, we show that PEPT2 associates with phagosome membranes. Together, these data identify Drosophila Yin and PEPT2 as evolutionarily conserved phagosome-associated transporters that are likely to be of particular importance in delivery of bacteria-derived ligands generated in phagosomes to cytosolic sensors recruited to the vicinity of these organelles.

Myeloperoxidase-dependent Inactivation of Surfactant Protein D in Vitro and in Vivo

Surfactant protein D (SP-D) plays diverse and important roles in innate immunity and pulmonary homeostasis. Neutrophils and myeloperoxidase (MPO) colocalized with SP-D in a murine bacterial pneumonia model of acute inflammation, suggesting that MPO-derived reactive species might alter the function of SP-D. Exposure of SP-D to the complete MPO-H(2)O(2)-halide system caused loss of SP-D-dependent aggregating activity. Hypochlorous acid (HOCl), the major oxidant generated by MPO, caused a similar loss of aggregating activity, which was accompanied by the generation of abnormal disulfide-cross-linked oligomers. A full-length SP-D mutant lacking N-terminal cysteine residues and truncation mutants lacking the N-terminal domains were resistant to the oxidant-induced alterations in disulfide bonding. Mass spectroscopy of HOCl-treated human SP-D demonstrated several modifications, but none involved key ligand binding residues. There was detectable oxidation of cysteine 15, but no HOCl-induced cysteine modifications were observed in the C-terminal lectin domain. Together, the findings localize abnormal disulfide cross-links to the N-terminal domain. MPO-deficient mice showed decreased cross-linking of SP-D and increased SP-D-dependent aggregating activity in the pneumonia model. Thus, MPO-derived oxidants can lead to modifications of SP-D structure with associated alterations in its characteristic aggregating activity.

A Novel Pseudopodial Component of the Dendritic Cell Anti-Fungal Response: The Fungipod

Fungal pathologies are seen in immunocompromised and healthy humans. C-type lectins expressed on immature dendritic cells (DC) recognize fungi. We report a novel dorsal pseudopodial protrusion, the “fungipod”, formed by DC after contact with yeast cell walls. These structures have a convoluted cell-proximal end and a smooth distal end. They persist for hours, exhibit noticeable growth and total 13.7±5.6 µm long and 1.8±0.67 µm wide at the contact. Fungipods contain clathrin and an actin core surrounded by a sheath of cortactin. The actin cytoskeleton, but not microtubules, is required for fungipod integrity and growth. An apparent rearward flow (225±55 nm/second) exists from the zymosan contact site into the distal fungipod. The phagocytic receptor Dectin-1 is not required for fungipod formation, but CD206 (Mannose Receptor) is the generative receptor for these protrusions. The human pathogen Candida parapsilosis induces DC fungipod formation strongly, but the response is species specific since the related fungal pathogens Candida tropicalis and Candida albicans induce very few and no fungipods, respectively. Our findings show that fungipods are dynamic actin-driven cellular structures involved in fungal recognition by DC. They may promote yeast particle phagocytosis by DC and are a specific response to large (i.e., 5 µm) particulate ligands. Our work also highlights the importance of this novel protrusive structure to innate immune recognition of medically significant Candida yeasts in a species specific fashion.

The Rap GTPases Regulate B Cell Morphology, Immune-Synapse Formation, and Signaling by Particulate B Cell Receptor Ligands

B lymphocytes spread and extend membrane processes when searching for antigens and form immune synapses upon contacting cells that display antigens on their surface. Although these dynamic morphological changes facilitate B cell activation, the signaling pathways underlying these processes are not fully understood. We found that activation of the Rap GTPases was essential for these changes in B cell morphology. Rap activation was important for B cell receptor (BCR)- and lymphocyte-function-associated antigen-1 (LFA-1)-induced spreading, for BCR-induced immune-synapse formation, and for particulate BCR ligands to induce localized F-actin assembly and membrane-process extension. Rap activation and F-actin assembly were also required for optimal BCR signaling in response to particulate antigens but not soluble antigens. Thus by controlling B cell morphology and cytoskeletal organization, Rap might play a key role in the activation of B cells by particulate and cell-associated antigens.

The human macrophage mannose receptor is not a professional phagocytic receptor

The macrophage mannose receptor (MR) appears to play an important role in the binding and phagocytosis of several human pathogens, but its phagocytic property and signaling pathways have been poorly defined. The general strategy to explore such topics is to express the protein of interest in nonphagocytic cells, but in the case of MR, there are few reports using the full-length MR cDNA. When we searched to clone de novo the human MR (hMR) cDNA, problems were encountered, and full-length hMR cDNA was only obtained after devising a complex cloning strategy. Chinese hamster ovary cells, which have a fully functional phagocytic machinery when expressing professional phagocytic receptors, were stably transfected, and cell clones expressing hMR at quantitatively comparable levels than human macrophages or J774E cells were obtained. They exhibited a functional hMR-mediated endocytic capacity of a soluble ligand but failed to ingest classical particulate ligands of MR such as zymosan, Mycobacterium kansasii, or trimannoside bovine serum albumin-coated latex beads. Transient expression of hMR in two human cell lines did not provide a phagocytic capacity either. In conclusion, we show that MR is not a professional phagocytic receptor, as it does not possess the ability to promote particle ingestion in nonphagocytic cells on its own. We propose that MR is a binding receptor, which requires a partner to trigger phagocytosis in some specialized cells such as macrophages. Our new expression vector could represent a useful tool to study the receptor and its partnership further.

High-resolution coral records of rare earth elements in coastal seawater: biogeochemical cycling and a new environmental proxy

In this study we have used laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), to produce a high resolution coral record of rare earth elements (REE), Mn and Ba from coastal Porites corals from the Great Barrier Reef of Australia. Validation of the LA-ICP-MS technique indicated that the method provides accurate and reproducible (RSD = 13–18%) analysis of low concentration REE in corals (∼1 to 100 ppb). The REE composition in coral samples was found to closely reflect that of the surrounding seawater and distribution coefficients of ∼1–2 indicated minimal fractionation of the series during incorporation into coral carbonate. To explore the idea that coral records of REE can be used to investigate dissolved seawater composition, we analyzed two coastal corals representing a total of ∼30 yr of growth, including a 10-yr overlapping period. Comparable results were obtained from the two samples, particularly in terms of elemental ratios (Nd/Yb) and the Ce anomaly. Based on this evidence and results from the determination of distribution coefficients, we suggest that useful records of seawater REE composition can be obtained from coral carbonates. When compared to the REE composition of a mid shelf coral, coastal corals showed a significant terrestrial influence, characterized by higher REE concentrations (greater than 10 times) and light REE enrichment. The REE composition of coastal seawater inferred from the coral record was dependent on seasonal factors and the influence of flood waters. REE fractionation displayed a strong seasonal cycle that correlated closely with Mn concentration. We suggest that higher Nd/Yb ratios and higher Mn concentrations in summer result from scavenging of heavy REE by particulate organic ligands and Mn reductive dissolution respectively, both processes displaying higher rates during periods of high primary productivity. The Ce anomaly also displayed a strong seasonal cycle showing an enhanced anomaly during summer and during flood events. This is consistent with the Ce anomaly being primarily controlled by the abundance of Ce oxidizing bacteria. Based on these arguments, we suggest that the coral record of dissolved REE and Mn may be regarded as a useful proxy for biological activity in coastal seawater.

Inverse dependency of particle residence times in ponds to the concentration of phosphate, the limiting nutrient

234Th, a commonly used short-lived particle-reactive tracer in marine systems, was measured in three different holding pond series at the Rocky Flats Environmental Technology Site (RFETS), Colorado, along with its parent nuclide 238U, to determine steady-state residence times of particle-reactive actinides such as Pu, and of particles. Series B ponds, which received industrial effluent that includes ortho-phosphate (PO 4) and actinides, differed from series A and C ponds, which did not. This difference was also evident in the calculated particle residence times, which were <1 day for the ponds B4 and B5, where PO 4 concentrations were higher (1.4 and 1.8 mg/l), and 3 and 3.4 days for ponds A3 and C2, respectively, where ortho-phosphate concentrations were lower (<0.1 mg/l). Particle residence times thus showed an inverse relationship with the concentration of ortho-phosphate, the limiting nutrient in fresh water systems. The same relationship to the concentration of ortho-phosphate or any of the other nutrient elements was not evident for the residence times of dissolved 234Th, which ranged between 0.1 and 2 days. This can be attributed to higher concentrations of dissolved and particulate ligands with greater binding potential for actinides such as four-valent Th and Pu in ponds with higher ortho-phosphate concentrations. Regardless of actual ortho-phosphate concentration, however, at water residence (holding) times of 1 month in these ponds, particles and associated actinides would be expected to be completely removed from the pond water to sediments.

Release of cadmium in the Danube estuary: contribution of physical and chemical processes as determined by an experimental approach

The behavior of dissolved cadmium (Cd) in the Danube estuary was investigated through field sampling and mixing experiments using Danube River water and Black Sea water. The experiments were performed by mixing these two end-member waters in various proportions, with the addition of stable or radioactive Cd to the freshwater Danube end-member prior to the mixing. The release of Cd that resulted in maximum concentrations under field conditions was well simulated by mixing experiments. The experimental results were modeled assuming that the release of Cd was the sum of the contribution of physical effects resulting from dilution effects and the contribution of chemical effects resulting from dissolved Cd-complex formation (and isotopic exchange when concerned). In the absence of dissolved Cd-complexing ligands, the release of Cd due to the dilution of the particulate phase during mixing could explain part of the maximum concentrations observed in field conditions. Kinetic effects were established by comparing the theoretical and measured contribution of chemical effects resulting from dissolved Cd-complex formation. The non-equilibrium state observed during the mixing experiment suggested the presence of particulate labile Cd that was not easily mobilized. All these features supported the hypothesis that Cd released in estuaries is controlled both by the dilution of the particulate phase and by kinetic competitive complexation between particulate ligands (covering a large spectrum of nature and strength) and dissolved ligands.

Distinct Roles for Lung Collectins in Pulmonary Host Defense

It has long been recognized that the lung contains resident components of the innate immune system that provide a first-line defense against infectious challenge. Recent studies have provided considerable evidence that two surfactant associated proteins, surfactant protein (SP)-A and SP-D, play important roles in pulmonary host defense (1). SP-A and SP-D are members of a larger family of proteins known as collectins (collagenous C-type lectins), which also includes the serum collectins mannose-binding lectin (MBL) and conglutinin (2). Members of this family are characterized by common N-terminal collagen-like domains and C-terminal carbohydrate recognition domains (CRD). They have the capacity to bind a variety of macromolecules, including carbohydrates, phospholipids, and proteins. Numerous in vitro and in vivo studies during the past decade have supported the hypothesis that SP-A and SP-D contribute to the containment of infections caused by a wide variety of respiratory pathogens, including bacteria, viruses, and fungi, through modulation of pathogen clearance, immune cell function, and lung inflammation (reviewed in Refs. 1–5). Similar effects have been noted for SP-A and SP-D in the innate immune response. However, specific differences in function have suggested that each collectin may play a very specific role in defense against different microbial pathogens. Although both SP-A and SP-D can bind to, aggregate, and/or promote uptake of certain pathogens by immune cells in the airway (summarized in Table 1), each collectin behaves in a very specific manner, which may define the outcome of infection. For example, SP-A has been shown to function as an efficient opsonin for a variety of bacteria such as Pseudomonas aeruginosa and mycobacteria (Table 1; reviewed in Refs. 3–7), and promote uptake of the complexes by alveolar macrophages. In contrast, SP-D is not a very effective opsonin, and only moderately enhances the phagocytosis of P. aeurginosa by alveolar macrophages (8), and inhibits phagocytosis of Mycobacterium tuberculosis (9). SP-A also enhances the phagocytic capacity of macrophages (10), and increases ingestion of nonopsonized bacteria such as M. tuberculosis (11) and Klebsiella (12). Although SP-A has been reported to promote aggregation of certain viruses and bacteria, SP-D forms large aggregates compared with the microscopic aggregates in the presence of SP-A. These large SP-D-associated aggregates may promote airway mucociliary clearance as well as internalization by pulmonary phagocytic cells (3). The differences in the oligomeric structures of these collectins may contribute to their interaction with various microbial pathogens. SP-A and SP-D are both assembled as oligomers of trimeric subunits (1, 3). Each subunit contains a short amino-terminal disulfide crosslinking domain, a triple helical collagen domain, a short trimeric coiled-coil linking domain, and a C-terminal CRD. SP-A exists predominantly as octadecamers in “bouquet-like” oligomers, with closely spaced CRDs. SP-D is assembled as dodecamers with long crosslinking domains, resulting in more widely spaced CRDs. Because the separation of CRDs in SP-D is fivefold greater than for SP-A (100 nm versus 20 nm), SP-D has a greater capacity to link interactions between binding sites on different particulate ligands, potentially contributing to the higher levels of aggregation of microbial ligands seen with SP-D. Key recent studies using mice deficient in SP-A or SP-D support a role for these collectins in pulmonary host defense (summarized in Table 2). Work from several groups using SP-A / mice has shown increased bacterial load following infection with group B streptococcus (13), and defective clearance of Hemophilus influenzae (14), P. aeruginosa (15), and Mycoplasma pulmonis (16). SP-A / mice also show increased susceptibility to challenge with respiratory syncyticial virus (RSV) (17). In most instances, the decreased microbial clearance can be reversed by addition of exogenous SP-A. Because these mice show normal respiratory function and surfactant lipid metabolism (20), the defects in microbial defense appear to be primarily attributable to SP-A. Mice lacking SP-A also show variable increases in proinflammatory mediators and decreases in anti-inflammatory cytokines, with an overall net proinflammatory environment. In contrast, evaluation of the role of SP-D using SP-D–deficient mice has been complicated by abnormal surfactant homeostasis and altered alveolar macrophage function (21). Nevertheless, in the absence of SP-D, several studies have examined infection of SP-D / mice with bacterial and viral pathogens, including group B streptococcus (14), H. influenzae (14), and influenza A virus (IAV) (19). Several differences in response to in vivo challenges with these microbes by SP-A / and SP-D / mice have been reported. First, SP-D / mice show no change in bacterial load following group B streptococcus and H. influenzae challenge compared with reduced clearance in SP-A–deficient mice (14). Second, whereas production of reactive oxygen species (ROS) by alveolar macrophages ( Received in original form January 7, 2002 )