Toxic and transcriptional responses of PC12 cells to soluble tungsten alloy surrogates.

There is increasing evidence that metals have a role in the etiology of diverse neurological diseases. This study used PC12 cells as an in vitro model to examine the toxicity of tungsten alloys that have important military applications. Initially, the relative concentrations of tungsten (W), nickel (Ni), and cobalt (Co) mobilized from pellets of a weapons-grade tungsten alloy incubated in physiologically relevant solutions were determined. Dosing solutions of soluble metal salts that were equivalent in ratio to those mobilized from these alloy pellets were used to treat nerve growth factor (NGF) differentiated PC12 cells. Treatments consisted of single (W, Ni or Co), paired (W/Ni, W/Co or Ni/Co) or complete (W/Ni/Co) metal exposures for 24 h followed by measurement of cytotoxicity, viability, and microarray analysis to examine their impact on survival and viability, global gene expression, and biological processes. Gene expression changed dramatically with addition of NGF. Addition of Ni or Co either singly or in combination further impacted gene expression. An observed additive effect of Ni and Co on gene expression was unaffected by the addition of W. The work showed that tungsten, as found in this tungsten alloy, had minimal relative toxicity as compared to the other alloy components when used either alone or in combination.

Assessment of 3-nitro-1,2,4-triazol-5-one as a potential endocrine disrupting chemical in rats using the Hershberger and uterotrophic bioassays.

The explosive 3-nitro-1,2,4-triazol-5-one (NTO) is an insensitive formulation developed to replace high energetics that are susceptible to accidental detonation from heat, shock, and impact. Although studies have shown NTO to be nontoxic at acute exposures, recent subacute and subchronic tests have demonstrated effects on testes and subsequent sperm production in rats. This study assessed endocrine disruption as a potential mechanism for these reproductive effects via the Hershberger and uterotrophic bioassays. These assays are 2 of the US Environmental Protection Agency's tier 1 in vivo screens for the Endocrine Disruptor Screening Program that measure differences in androgen- and estrogen-sensitive tissue weights in castrated and ovariectomized rats. The gonadectomized rats were orally exposed to NTO in a corn oil vehicle at doses of 250, 500, or 1000 mg/kg body weight (bw)/d for 10 and 3 days for the Hershberger and uterotrophic assays, respectively, according to standard protocols. Male rats also received testosterone (0.2 mg/kg/d, subcutaneous) and antiandrogenic flutamide (3mg/kg/d, oral) as negative and positive controls, and females received 17 α-ethynyl estradiol (0.3 µg/d, subcutaneous) as positive controls. 3-Nitro-1,2,4-triazol-5-one caused neither a decrease in androgen-sensitive male reproductive selected tissue (seminal vesicles with fluid/without fluid, glans penis, Cowper gland, ventral prostrate, and levator ani-bulbocavernosus) weights nor a change in uterine weights. The results of this study provide no evidence to suggest that NTO acts like an estrogenic or antiandrogenic endocrine disruptor in rats at these doses.

In vivo corrosion, tumor outcome, and microarray gene expression for two types of muscle-implanted tungsten alloys.

Tungsten alloys are composed of tungsten microparticles embedded in a solid matrix of transition metals such as nickel, cobalt, or iron. To understand the toxicology of these alloys, male F344 rats were intramuscularly implanted with pellets of tungsten/nickel/cobalt, tungsten/nickel/iron, or pure tungsten, with tantalum pellets as a negative control. Between 6 and 12 months, aggressive rhabdomyosarcomas formed around tungsten/nickel/cobalt pellets, while those of tungsten/nickel/iron or pure tungsten did not cause cancers. Electron microscopy showed a progressive corrosion of the matrix phase of tungsten/nickel/cobalt pellets over 6 months, accompanied by high urinary concentrations of nickel and cobalt. In contrast, non-carcinogenic tungsten/nickel/iron pellets were minimally corroded and urinary metals were low; these pellets having developed a surface oxide layer in vivo that may have restricted the mobilization of carcinogenic nickel. Microarray analysis of tumors revealed large changes in gene expression compared with normal muscle, with biological processes involving the cell cycle significantly up-regulated and those involved with muscle development and differentiation significantly down-regulated. Top KEGG pathways disrupted were adherens junction, p53 signaling, and the cell cycle. Chromosomal enrichment analysis of genes showed a highly significant impact at cytoband 7q22 (chromosome 7) which included mouse double minute (MDM2) and cyclin-dependant kinase (CDK4) as well as other genes associated with human sarcomas. In conclusion, the tumorigenic potential of implanted tungsten alloys is related to mobilization of carcinogenic metals nickel and cobalt from corroding pellets, while gene expression changes in the consequent tumors are similar to radiation induced animal sarcomas as well as sporadic human sarcomas.

The role of anion exchange in the uptake of Pb by human erythrocytes and Madin-Darby canine kidney cells.

Anion exchange (AE) plays a critical role in regulating intracellular pH in erythrocytes and epithelial cells and has been suggested to facilitate the transport of lead (Pb) across the erythrocyte cell membrane. In this study we examined the role of AE in the uptake of Pb by human erythrocytes and by Madin-Darby canine kidney (MDCK) cells, the kidney epithelial cell line. Functional AE in MDCK cells was evidenced by: increased uptake of SO(4)(2-) at pH 6.0 over pH 7.0, and inhibition of SO(4)(2-) uptake by the AE inhibitor 4, 4'-diisothiocyanostilbene-2, 2'- disulfonic acid (DIDS) as well as by non-halide anions. Accumulation of Pb into MDCK cells was time and temperature dependent. DIDS inhibited uptake of Pb into human erythrocytes but not MDCK cells. In conclusion, uptake of Pb into erythrocytes but not kidney epithelial cells occurs through AE.

Pilot study of sources of lead exposure in Moscow, Russia.

This preliminary investigation of sources of lead exposure in Moscow, Russia, by Russian and US collaborators measured lead in paint, interior dust, and drinking water in seven day-care centres, and in petrol, soil and canned food. Some paint samples exceeded US regulatory standards for lead in paint on surfaces (0.5%). Dust lead loadings were < 1.7 µg cm(-2) and below the guidance levels of the US EPA. Drinking water lead concentrations were at or below the US drinking water standard of 15 µg L(-1). Lead concentrations in petrol from Moscow vehicles and petrol stations were consistent with a regulation banning the sale of leaded petrol within the Moscow City limits. Except for baby food, lead levels were higher in the Russian canned foods (range 6 to 1240 µg kg(-1), dry weight) compared to corresponding US canned foods, with ratios of Russian to US levels of up to 120:1 for evaporated milk. Lead concentrations in soil generally ranged from 500 to 2000 µg g(-1), levels that would trigger hazard reduction measures according to US EPA guidance. These findings, together with the use of lead in petrol outside Moscow, indicate multiple sources of lead exposure in Russia. Priorities for future research are discussed including the establishment of interlaboratory quality control programmes.

Graphite furnace atomic absorption spectroscopic measurement of blood lead in matrix-matched standards.

Now that the level of concern for a toxic blood lead concentration is 0.482 mumol/L (10 micrograms/dL), laboratories must meet new requirements to shorten analysis times and increase accuracy and precision of blood lead determinations. We used a matrix-matching method to estimate the lead concentration in blood by graphite furnace atomic absorption spectroscopy (GFAAS). For CDC proficiency samples and the NIST-Certified Blood Reference standard, the performance of this method compared favorably with that of previously published GFAAS methods and of the anodic stripping voltammetric method routinely used in our laboratory. At lead concentrations of 0.242 mumol/L (5.01 micrograms/dL) and 1.478 mumol/L (30.63 micrograms/dL), within-run CVs were 2.78% and 0.68%, respectively; between-run CVs were 4.9% and 1.35%. In 52 study samples with lead content ranging from 0.097 to 3.812 mumol/L (2 to 79 micrograms/dL), 87% of results by the matrix-modified method were within 0.048 mumol/L (1 microgram/dL) of consensus values.