Fe- and Cu-complex formation with artificial ligands investigated by ultra-high resolution Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS): Implications for natural metal-organic complex studies

In recent years, electrospray-ionization mass spectrometry (ESI-MS) has been increasingly used to complement the bulk determination of metal-ligand equilibria, for example via competitive ligand exchange-adsorptive cathodic stripping voltammetry (CLE-ACSV). However, ESI-MS speciation analyses may be impacted by instrumental artifacts such as reduction reactions, fragmentation, and adduct formation at the ESI source, changes in the ionization efficiencies of the detected species in relation to sample matrix, and peak overlaps in response to increasing sample complexity. In our study, equilibria of the known artificial ligands citrate, ethylenediaminetetraacetic acid (EDTA), 1-nitroso-2-naphthol (NN), and salicylaldoxime (SA) with iron (Fe) and copper (Cu) were investigated by ultra-high resolution ESI-MS, Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS), under a variety of sample matrix and ionization settings. The acquired mass spectra were compared with metal-ligand equilibrium data from the literature as well as an adapted speciation model. Overall, the mass spectra produced representative species mentioned in previous reports and predicted by the speciation calculations, such as Fe(Cit), Cu(Cit)2, Fe(EDTA), Cu(EDTA), Fe(NN)3, and Cu(SA)2. The analyses furthermore revealed new species which had been hypothesized but not measured directly using other methods, for example ternary complexes of citrate with Fe and Cu, Cu(SA) monomers, and the dimer Fe(SA)2. Finally, parallel measurements of a Cu+SA calibration series and a Cu+SA+EDTA competition series indicated that FT-ICR-MS can produce linear responses and low detection limits analogous to those of ACSV. We propose that ultra-high resolution FT-ICR-MS can be used as a representative tool to study interactions of trace metals with artificial as well as natural, unknown ligands at the molecular level

High-Level Expression of Various Apolipoprotein (a) Isoforms by "Transferrinfection". The Role of Kringle IV Sequences in the Extracellular Association with Low-Density Lipoprotein

Characterization of the assembly of lipoprotein(a) [Lp(a)] is of fundamental importance to understanding the biosynthesis and metabolism of this atherogenic lipoprotein. Since no established cell lines exist that express Lp(a) or apolipoprotein(a) [apo(a)], a "transferrinfection" system for apo(a) was developed utilizing adenovirus receptor- and transferrin receptor-mediated DNA uptake into cells. Using this method, different apo(a) cDNA constructions of variable length, due to the presence of 3, 5, 7, 9, 15, or 18 internal kringle IV sequences, were expressed in cos-7 cells or CHO cells. All constructions contained kringle IV-36, which includes the only unpaired cysteine residue (Cys-4057) in apo(a). r-Apo(a) was synthesized as a precursor and secreted as mature apolipoprotein into the medium. When medium containing r-apo(a) with 9, 15, or 18 kringle IV repeats was mixed with normal human plasma LDL, stable complexes formed that had a bouyant density typical of Lp(a). Association was substantially decreased if Cys-4057 on r-apo(a) was replaced by Arg by site-directed mutagenesis or if Cys-4057 was chemically modified. Lack of association was also observed with r-apo(a) containing only 3, 5, or 7 kringle IV repeats without "unique kringle IV sequences", although Cys-4057 was present in all of these constructions. Synthesis and secretion of r-apo(a) was not dependent on its sialic acid content. r-Apo(a) was expressed even more efficiently in sialylation-defective CHO cells than in wild-type CHO cells. In transfected CHO cells defective in the addition of N-acetylglucosamine, apo(a) secretion was found to be decreased by 50%. Extracellular association with LDL was not affected by the carbohydrate moiety of r-apo(a), indicating a protein-protein interaction between r-apo(a) and apoB. These results show that, besides kringle IV-36, other kringle IV sequences are necessary for the extracellular association of r-apo(a) with LDL. Changes in the carbohydrate moiety of apo(a), however, do not affect complex formation

Riverine mixing at the molecular scale - An ultrahigh-resolution mass spectrometry study on dissolved organic matter and selected metals in the Amazon confluence zone (Manaus, Brazil)

The Amazon delivers a fifth of the global continental runoff and riverine dissolved organic carbon (DOC) to the ocean. Intensified biogeochemical processes are expected at the junction of the Amazon's major blackwater tributary, the Rio Negro, and its parent, the Rio Solimbes, due to large gradients in pH, conductivity, DOC and particle load. Dissolved organic matter (DOM) plays a major role in aquatic biogeochemical processes which are poorly understood on the molecular level. To gain insights into the potential role of DOM in non-conservative processes, we assessed dynamics of Cu, Fe and DOM by ultrahigh resolution mass spectrometry in: (1) endmembers, (2) regional samples and (3) laboratory mixing experiments under presence/absence of natural particles (>0.2 p.m). The relative abundances of 3600 DOM molecular formulae were interpreted via multivariate statistics which revealed major dynamics in the DOM molecular composition. >40% of molecular formulae displayed conservative behavior even in the presence of natural particles, agreeing with bulk DOC behavior, but opposing the oftenpresumed non-conservative behavior of DOM. Another 16-27% of formulae fluctuated in FT-ICRMS signal intensity during mixing, but did not show consistent non-conservative behavior. Both rivers left a clear molecular imprint within the DOM of the Amazon, each being linked to >800 molecular formulae. Characteristic for the Rio Negro was a dominance of phenolics with a wide molecular mass range (centered at similar to 400 Da), and for the Rio Solimbes more saturated but lower-molecular mass compounds (centered at similar to 300 Da). Both Fe and Cu showed distinct non-conservative mixing patterns under particle presence. In the controlled mixing experiments including original particles at natural concentration, up to 0.5 ttg/L Cu was released from the particles into solution at 20-40% blackwater contribution. Our molecular analysis revealed distinct DOM compositional changes in polyphenol- and nitrogencontaining formulae paralleling this release, suggesting links to desorption of potential ligands and charge-induced effects at particle surfaces caused by pH and conductivity changes in the course of mixing. (C) 2019 Elsevier Ltd. All rights reserved

Lipoprotein(a) is associated differentially with carotid stenosis, occlusion, and total plaque area

Background - Lipoprotein(a) [Lp(a)] is a putative risk factor for myocardial infarction and stroke and is related to thrombosis and impaired fibrinolysis. We studied relationships of Lp(a) with carotid stenosis, occlusion, and total plaque area, distinct phenotypes of atherosclerosis that may be differentially affected by cardiovascular risk factors. Methods and Results - Multivariable linear regression analysis was used to study relationships of Lp(a) to phenotypes of carotid atherosclerosis among 876 consecutive patients from an atherosclerosis prevention clinic with complete data for all variables used in the model. Occlusion of an internal carotid artery was present in 22 (2.5%) patients (one with bilateral occlusions). Risk factors predicted carotid plaque area, stenosis, and occlusion differently. Lp(a) was a significant independent predictor of baseline stenosis (P\u3c0.0001) but not of plaque area (P=0.13); in logistic regression, Lp(a) significantly predicted occlusion (P=0.001). Patients with occlusion had significantly higher levels of Lp(a): 0.27±0.25 g/L versus 0.17±0.18 g/L without occlusion; P=0.007. Conclusion - Lp(a) was a significant independent predictor of carotid stenosis and occlusion, but not of carotid plaque area, supporting the hypothesis that the effect of Lp(a) on atherogenesis and cardiovascular risk is largely related to thrombosis and impaired fibrinolysis. Stenosis and occlusion may not be attributable to plaque progression, but to plaque rupture and thrombosis; this relationship may also apply to other arterial beds. © 2008 American Heart Association, Inc

The Position of High Frequency Waves with Respect to the Granulation Pattern

High frequency velocity oscillations were observed in the spectral lines Fe I 543.45nm and 543.29nm, using 2D spectroscopy with a Fabry- Perot and speckle reconstruction, at the VTT in Tenerife. We investigate the radial component of waves with frequencies in the range 8 - 22mHz in the internetwork, network and a pore. We find that the occurrence of waves do not show any preference on location and are equally distributed over down-flows and up-flows, regardless of the activity of the observed area in the line of Fe I 543.45nm. The waves observed in the lower formed line of Fe I 543.29nm seem to appear preferentially over down-flows.Comment: Article has 12 pages and 7 images. It is accepted in Solar Physics Journa

Diking, young volcanism and diffuse hydrothermal activity on the southern Mid-Atlantic Ridge: The Lilliput field at 9°33'S

Detailed exploration with remotely operated and autonomous deep submergence vehicles has revealed, at 9 degrees 33'S, the presence of the southernmost active hydrothermal field known so far on the Mid-Atlantic Ridge. The size of the hydrothermal field, which we have named "Lilliput", is about 1000 m x 250 m. It lies in a water depth of 1500 m on a ridge segment (Segment A3) with considerably thickened crust of 11 km. Four relatively small diffuse vent sites occur on a large young (estimated <100 years old) lava flow, partly covering the flow with hydrothermal Fe-oxide/hydroxide sediments. Based on homogeneous major element compositions of ca. 25 lava samples, this flow covers an area of at least 5 km x 0.6 km. The lava flow erupted from a series of parallel fissures at the western edge of the flow and a volcanic ridge consisting of up to 30 m high pillow mounds. The volcanic ridge probably represents the surface expression of an underlying dike which fed the flow. Several drained lava pond structures were observed within the flow but only one shows hydrothermal activity. The hydrothermal venting and precipitation of abundant Fe-oxyhydroxides appear to be related to the young diking and eruption event and the four different hydrothermally active sites of the Lilliput field lie along and almost equidistant from the eastern flank of the supposed dike. Although a hydrothermal plume some 500 m above the seafloor was found in two consecutive years (2005 and 2006), no high-temperature venting associated with Lilliput has been found. in agreement with findings at other ridges with thick crust such as Reykjanes. High magma supply rate and frequent diking and eruption events may lead to hot hydrothermal vents being rare in slow-spreading segments with thick crust whereas diffuse venting is abundant. Interestingly, the fauna at the Lilliput vents largely consists of small and apparently juvenile mussels (Bathymodiolus sp.) and did not show any signs of growth during the four years of continuing observations possibly reflecting pulsing hydrothermal activity

Small-scale heterogeneity of trace metals including REY in deep-sea sediments and pore waters of the Peru Basin, southeastern equatorial Pacific

Due to its remoteness, the deep-sea floor remains an understudied ecosystem of our planet. The patchiness of existing data sets makes it difficult to draw conclusions about processes that apply to a wider area. In our study we show how different settings and processes determine sediment heterogeneity on small spatial scales. We sampled solid phase and porewater from the upper 10 m of an approximately 7.4×13 km2 area in the Peru Basin, in the southeastern equatorial Pacific Ocean, at 4100 m water depth. Samples were analyzed for trace metals, including rare earth elements and yttrium (REY), as well as for particulate organic carbon (POC), CaCO3, and nitrate. The analyses revealed the surprisingly high spatial small-scale heterogeneity of the deep-sea sediment composition. While some cores have the typical green layer from Fe(II) in the clay minerals, this layer is missing in other cores, i.e., showing a tan color associated with more Fe(III) in the clay minerals. This is due to varying organic carbon contents: nitrate is depleted at 2–3 m depth in cores with higher total organic carbon contents but is present throughout cores with lower POC contents, thus inhibiting the Fe(III)-to-Fe(II) reduction pathway in organic matter degradation. REY show shale-normalized (SN) patterns similar to seawater, with a relative enrichment of heavy REY over light REY, positive LaSN anomaly, negative CeSN anomaly, and positive YSN anomaly and correlate with the Fe-rich clay layer and, in some cores, also correlate with P. We therefore propose that Fe-rich clay minerals, such as nontronite, as well as phosphates, are the REY-controlling phases in these sediments. Variability is also seen in dissolved Mn and Co concentrations between sites and within cores, which might be due to dissolving nodules in the suboxic sediment, as well as in concentration peaks of U, Mo, As, V, and Cu in two cores, which might be related to deposition of different material at lower-lying areas or precipitation due to shifting redox boundaries

Copper-nickel-rich, amalgamated ferromanganese crust-nodule deposits from Shatsky Rise, NW Pacific

A unique set of ferromanganese crusts and nodules collected from Shatsky Rise (SR), NW Pacific, were analyzed for mineralogical and chemical compositions, and dated using Be isotopes and cobalt chronometry. The composition of these midlatitude, deep-water deposits is markedly different from northwest-equatorial Pacific (PCZ) crusts, where most studies have been conducted. Crusts and nodules on SR formed in close proximity and some nodule deposits were cemented and overgrown by crusts, forming amalgamated deposits. The deep-water SR crusts are high in Cu, Li, and Th and low in Co, Te, and Tl concentrations compared to PCZ crusts. Thorium concentrations (ppm) are especially striking with a high of 152 (mean 56), compared to PCZ crusts (mean 11). The deep-water SR crusts show a diagenetic chemical signal, but not a diagenetic mineralogy, which together constrain the redox conditions to early oxic diagenesis. Diagenetic input to crusts is rare, but unequivocal in these deep-water crusts. Copper, Ni, and Li are strongly enriched in SR deep-water deposits, but only in layers older than about 3.4 Ma. Diagenetic reactions in the sediment and dissolution of biogenic calcite in the water column are the likely sources of these metals. The highest concentrations of Li are in crust layers that formed near the calcite compensation depth. The onset of Ni, Cu, and Li enrichment in the middle Miocene and cessation at about 3.4 Ma were accompanied by changes in the deep-water environment, especially composition and flow rates of water masses, and location of the carbonate compensation depth. Key Points - Fe-Mn crusts can have a diagenetic component - Mid-latitude N. Pacific deep-water Fe-Mn crusts are uniquely enriched in Cu, Th, Li - Temporal changes in deep-ocean geochemical processes are recorde