Michael E. Ketterer

Role of Ceruloplasmin in Macrophage Iron Efflux during Hypoxia

The reticuloendothelial system has a central role in erythropoiesis and iron homeostasis. An important function of reticuloendothelial macrophages is phagocytosis of senescent red blood cells. The iron liberated from heme is recycled for delivery to erythrocyte precursors for a new round of hemoglobin synthesis. The molecular mechanism by which recycled iron is released from macrophages remains unresolved. We have investigated the mechanism of macrophage iron efflux, focusing on the role of ceruloplasmin (Cp), a copper protein with a potent ferroxidase activity that converts Fe2+ to Fe3+ in the presence of molecular oxygen. As shown by others, Cp markedly increased iron binding to apotransferrin at acidic pH; however, the physiological significance of this finding is uncertain because little stimulation was observed at neutral pH. Introduction of a hypoxic atmosphere resulted in marked Cp-stimulated binding of iron to apotransferrin at physiological pH. The role of Cp in cellular iron release was examined in U937 monocytic cells induced to differentiate to the macrophage lineage. Cp added at its normal plasma concentration increased the rate of 55Fe release from U937 cells by about 250%. The stimulation was absolutely dependent on the presence of apotransferrin and hypoxia. Cp-stimulated iron release was confirmed in mouse peritoneal macrophages. Stimulation of iron release required an intracellular “labile iron pool” that was rapidly depleted in the presence of Cp and apotransferrin. Ferroxidase-mediated loading of iron into apotransferrin was critical for iron release because ferroxidase-deficient Cp was inactive and because holotransferrin could not substitute for apotransferrin. The extracellular iron concentration was critical as shown by inhibition of iron release by exogenous free iron, and marked enhancement of release by an iron chelator. Together these data show that Cp stimulates iron release from macrophages under hypoxic conditions by a ferroxidase-dependent mechanism, possibly involving generation of a negative iron gradient.

Anticancer Activity of Sodium Stibogluconate in Synergy with IFNs

Cancer cell resistance limits the efficacy of IFNs. In this study, we show that sodium stibogluconate (SSG) and IFN-α synergized to overcome IFN-α resistance in various human cancer cell lines in culture and eradicated IFN-α-refractory WM9 human melanoma tumors in nude mice with no obvious toxicity. SSG enhanced IFN-α-induced Stat1 tyrosine phosphorylation, inactivated intracellular SHP-1 and SHP-2 that negatively regulate IFN signaling, and induced cellular protein tyrosine phosphorylation in cancer cell lines. These effects are consistent with inactivation of phosphatases as the basis of SSG anticancer activity. Characterization of SSG by chromatography revealed that only selective compounds in SSG were effective protein tyrosine phosphatase inhibitors. These observations suggest the potential of SSG as a clinically usable protein tyrosine phosphatase inhibitor in cancer treatment and provide insights for developing phosphatase-targeted therapeutics.

Resolving global versus local/regional Pu sources in the environment using sector ICP-MS

Sector inductively coupled plasma mass spectrometry is a versatile method for the determination of plutonium activities and isotopic compositions in samples containing this element at fallout levels. Typical detection limits for 239 + 240Pu are 0.1, 0.02 and 0.002 Bq kg−1Pu for samples sizes of 0.5 g, 3 g, and 50 g of soil, respectively. The application of sector ICP-MS-based Pu determinations is demonstrated in studies in sediment chronology, soil Pu inventory and depth distribution, and the provenance of global fallout versus local or regional Pu sources. A sediment core collected from Sloans Lake (Denver, Colorado, USA) exhibits very similar 137Cs and 239 + 240Pu activity profiles; 240Pu/239Pu atom ratios indicate possible small influences from the Nevada Test Site and/or the Rocky Flats Environmental Technology Site. An undisturbed soil profile from Lockett Meadow (Flagstaff, Arizona, USA) exhibits an exponential decrease in 239 + 240Pu activity versus depth; 240Pu/239Pu in the top 3 cm is slightly lower than the global fallout range of 0.180 ± 0.014 due to possible regional influence of Nevada Test Site fallout. The 239 ± 240Pu inventory at Lockett Meadow is 56 ± 4 Bq m−2, consistent with Northern Hemisphere mid-latitude fallout. Archived NdF3 sources, prepared from Polish soils, demonstrate that substantial 239 + 240Pu from the 1986 Chernobyl disaster has been deposited in north eastern regions of Poland; compared to global fallout, Chernobyl Pu exhibits higher abundances of 240Pu and 241Pu. The ratios 240Pu/239Pu and 241Pu/239Pu co-vary and range from 0.186–0.348 and 0.0029–0.0412, respectively, in forest soils (241Pu/239Pu = 0.2407·[240Pu/239Pu] − 0.0413; r2 = 0.9924).

Suitability of 239+240Pu and 137Cs as tracers for soil erosion assessment in mountain grasslands.

Anthropogenic radionuclides have been distributed globally due to nuclear weapons testing, nuclear accidents, nuclear weapons fabrication, and nuclear fuel reprocessing. While the negative consequences of this radioactive contamination are self-evident, the ubiquitous fallout radionuclides (FRNs) distribution form the basis for the use as tracers in ecological studies, namely for soil erosion assessment. Soil erosion is a major threat to mountain ecosystems worldwide. We compare the suitability of the anthropogenic FRNs, 137Cs and 239+240Pu as soil erosion tracers in two alpine valleys of Switzerland (Urseren Valley, Canton Uri, Central Swiss Alps and Val Piora, Ticino, Southern Alps). We sampled reference and potentially erosive sites in transects along both valleys. 137Cs measurements of soil samples were performed with a Li-drifted Germanium detector and 239+240Pu with ICP-MS. Our data indicates a heterogeneous deposition of the 137Cs, since most of the fallout origins from the Chernobyl April/May 1986 accident, when large parts of the European Alps were still snow-covered. In contrast, 239+240Pu fallout originated mainly from 1950s to 1960s atmospheric nuclear weapons tests, resulting in a more homogenous distribution and thus seems to be a more suitable tracer in mountainous grasslands. Soil erosion assessment using 239+240Pu as a tracer pointed to a huge dynamic and high heterogeneity of erosive processes (between sedimentation of 1.9 and 7 t ha(-1) yr(-1) and erosion of 0.2-16.4 t ha(-1) yr(-1) in the Urseren Valley and sedimentation of 0.4-20.3 t ha(-1) yr(-1) and erosion of 0.1-16.4 t ha(-1) yr(-1) at Val Piora). Our study represents a novel and successful application of 239+240Pu as a tracer of soil erosion in a mountain environment.

Elevated rates of organic carbon, nitrogen, and phosphorus accumulation in a highly impacted mangrove wetland

The effect of nutrient enrichment on mangrove sediment accretion and carbon accumulation rates is poorly understood. Here we quantify sediment accretion through radionuclide tracers to determine organic carbon (OC), total nitrogen (TN), and total phosphorus (TP) accumulation rates during the previous 60 years in both a nutrient-enriched and a pristine mangrove forest within the same geomorphological region of southeastern Brazil. The forest receiving high nutrient loads has accumulated OC, TN, and TP at rates that are fourfold, twofold, and eightfold respectively, higher than those from the undisturbed mangrove. Organic carbon and TN stable isotopes (δ13C and δ15N) reflect an increased presence of organic matter (OM) originating with either phytoplankton, benthic algae, or another allochthonous source within the more rapidly accumulated sediments of the impacted mangrove. This suggests that the accumulation rate of OM in eutrophic mangrove systems may be enhanced through the addition of autochthonous and allochthonous nonmangrove material.

Childhood lead exposure and uptake in teeth in the Cleveland area during the era of leaded gasoline

Childhood uptake of lead from exposure to atmospheric leaded gasoline in the United States has been studied using mainly blood lead levels. Since reliable blood lead techniques were used only after the peak use of leaded gasoline, the prior exposure history is unclear. The well-documented decline in blood lead levels after the mid-1970s could represent the continuation of a historic steady decline in exposure from many sources. Alternatively, the post-1970s decline might represent the declining phase of a unimodal rise and fall corresponding closely to usage of leaded gasoline. To assess these possibilities, lead concentration and 207Pb/206Pb isotope ratios were measured in the enamel of permanent molar teeth formed between 1936 and 1993 in mainly African-American donors who grew up in the Cleveland area. Tooth enamel preserves the lead concentration and isotope ratio that prevails during tooth formation. Historical trends in enamel lead concentration were significantly correlated with surrogates of atmospheric lead exposure: lead in sediments of two dated Lake Erie cores, and lead consumed in gasoline. About two-thirds of the total lead uptake into enamel in this period was attributable to leaded gasoline, and the remainder to other sources (e.g. paint). Enamel 207Pb/206Pb isotope ratios were similar to those of one lake sediment. Multivariate analysis revealed significant correlation in neighborhoods with higher levels of traffic, and including lake sediment data, accounted for 53% of the variation in enamel lead levels. Enamel lead concentration was highly correlated with reported African-American childhood blood levels. The extrapolated peak level of 48microg/dL (range 40 to 63) is associated with clinical and behavioral impairments, which may have implications for adults who were children during the peak gasoline lead exposure. In sum, leaded gasoline emission was the predominant source of lead exposure of African-American Cleveland children during the latter two-thirds of the 20th century.

Lead isotopic and chalcophile element compositions in the environment near a zinc smelting-secondary zinc recovery facility, Palmerton, Pennsylvania, USA

Abstract The environment surrounding Palmerton, Pennsylvania is contaminated with Pb arising from primary Zn smelting and a process involving Zn recovery from electric arc steel furnace dusts. Lead isotope systematics have been used to distinguish primary Zn smelting Pb (206Pb/204Pb∼18.4–18.5) from electric arc furnace dust lead (206Pb/204Pb∼19.0–19.1). Primary Zn smelting is the dominant source of Pb in O2 horizon soils from undisturbed near-Palmerton locations, which contain up to 3570 ppm Pb and 782 ppm Cd. Soils from undeveloped near-Palmerton locations also exhibit unusually elevated concentrations of other sphalerite-derived chalcophilic elements (Se, Ag, In, Sb, Te, Au, Hg, Tl and Bi); indium concentrations of up to 17.0 ppm are observed therein. Residential soils and dusts from Palmerton contain Pb which is largely explainable via mixing of Pb from primary Zn smelting and electric arc furnace dusts. Approximately 80% of the Pb in airborne particulate matter sampled at Palmerton in 1991 is derived from electric arc furnace dusts, and atmospheric enrichment factors for Cu, Sb, Pb, and Bi are observed which confirm this major source contribution. Residential samples from a control location contain Pb which is less radiogenic than is found in Palmerton, and exhibit no unusual elevation in sphalerite elements. Lead source discrimination in the Palmerton environment via Pb isotopic and elemental constituents approaches result in parallel conclusions.

Examining 239+240Pu, 210Pb and historical events to determine carbon, nitrogen and phosphorus burial in mangrove sediments of Moreton Bay, Australia

Two sediment cores were collected in a mangrove forest to construct geochronologies for the previous century using natural and anthropogenic radionuclide tracers. Both sediment cores were dated using (239+240)Pu global fallout signatures as well as (210)Pb, applying both the Constant Initial Concentration (CIC) and the Constant Rate of Supply (CRS) models. The (239+240)Pu and CIC model are interpreted as having comparable sediment accretion rates (SAR) below an apparent mixed region in the upper ∼5 to 10 cm. In contrast, the CRS dating method shows high sediment accretion rates in the uppermost intervals, which is substantially reduced over the lower intervals of the 100-year record. A local anthropogenic nutrient signal is reflected in the high total phosphorus (TP) concentration in younger sediments. The carbon/nitrogen molar ratios and δ(15)N values further support a local anthropogenic nutrient enrichment signal. The origin of these signals is likely the treated sewage discharge to Moreton Bay which began in the early 1970s. While the (239+240)Pu and CIC models can only produce rates averaged over the intervals of interest within the profile, the (210)Pb CRS model identifies elevated rates of sediment accretion, organic carbon (OC), nitrogen (N), and TP burial from 2000 to 2013. From 1920 to 2000, the three dating methods provide similar OC, N and TP burial rates, ∼150, 10 and 2 g m(-2) year(-1), respectively, which are comparable to global averages.

Diatom changes in two Uinta mountain lakes, Utah, USA: responses to anthropogenic and natural atmospheric inputs

Diatom assemblages in sediments from two subalpine lakes in the Uinta Mountains, Utah, show asynchronous changes that are related to both anthropogenic and natural inputs of dust. These lakes are downwind of sources of atmospheric inputs originating from mining, industrial, urban, agricultural and natural sources that are distributed within tens to hundreds of kilometers west and south of the Uinta Mountains. Sediment cores were retrieved from Marshall and Hidden lakes to determine the impacts of atmospheric pollution, especially metals. Paleolimnological techniques, including elemental analyses and 210Pb and 239+240Pu dating, indicate that both lakes began receiving eolian inputs from anthropogenic sources in the late 1800s with the greatest increases occurring after the early 1900s. Over the last century, sediments in Marshall Lake, which is closer to the Wasatch Front and receives more precipitation than Hidden Lake, received twice the concentrations of metals and phosphorus as Hidden Lake. Comparison of diatom and elemental data reveals coeval changes in geochemistry and diatom assemblages at Marshall Lake, but not at Hidden Lake; however, a major shift in diatom assemblages occurs at Hidden Lake in the seventeenth century. The change in diatoms at Marshall Lake is marked by the near disappearance of Cyclotella stelligera and C. pseudostelligera and an increase in benthic, metal-tolerant diatoms. This change is similar to changes in other lakes that have been attributed to metal pollution. The marked change in diatom assemblages at Hidden Lake indicates a shift in lake-water pH from somewhat acidic to circumneutral. We hypothesize that this change in pH is related to drought-induced changes in input of carbonate-rich desert dust.

Applications of Transuranics as Tracers and Chronometers in the Environment

The transuranic elements (TRU) Np, Pu, Am, and Cm have prominently emerged as powerful tracers of earth and environmental processes, applicable to the recent, post nuclear-era timescale. Various long-lived isotopes of these elements are found in the earth’s surface environment, almost exclusively as a result of nuclear weapons production, testing, or nuclear fuel cycle activities. A globally recognizable signal, of consistent composition, from stratospheric fallout derived from 1950–1960 above-ground weapons tests is itself useful in tracing applications; in specific local/regional settings, stratospheric fallout is mixed with or dominated by other TRU sources with contrasting isotopic signatures. Both decay-counting and MS approaches have been utilized to measure the concentrations and isotopic ratios of TRU and are useful as discriminators for source characterization, provenance, and apportionment. Examples include the activity ratios 238Pu/239+240Pu, 241Am/239+240Pu, and 241Pu/239+240Pu; atom ratios such as 240Pu/239Pu, 237Np/239Pu, 241Pu/239Pu, and 242Pu/239Pu are also used in this context. Of the TRU elements, Pu is by far the most widely studied; accordingly, this chapter mainly emphasizes the use of Pu activities and/or atom ratios as tracers and/or chronometers. Nevertheless, Pu is sometimes measured in combination with one or more isotopes of other elements. The TRU elements offer several prominent applications in environmental/geochemical tracing: (1) chronostratigraphy of sediments and related recent Holocene deposits; (2) using fallout TRU as quantitative probes of soil erosion, transport and deposition; (3) investigating water mass circulation, the transport and scavenging of particulate matter, and tracking the marine geochemical behavior of the TRU elements themselves in the marine environment; and (4) studies of the local/regional transport, deposition and inventories of non-fallout TRU in the surficial environment.