The cytotoxicity and genotoxicity of particulate and soluble hexavalent chromium in human lung cells

Hexavalent chromium (Cr(VI)) is a human lung carcinogen. Cr(VI) is a particularly important and dangerous carcinogen, because there is widespread exposure to it both occupationally and to the general public. However, despite the potential for widespread exposure and the fact that the lung is its target organ, there are few reports of the genotoxicity of Cr(VI) in human lung cells. Clearly, in order to better understand this carcinogen, its effects in its target cells need to be evaluated. Accordingly, we determined the cytotoxicity and clastogenicity of both particulate (water-insoluble) and soluble Cr(VI) in primary human bronchial fibroblasts (PHBFs). We used lead chromate (PbCrO(4)) and sodium chromate (Na(2)CrO(4)) as prototypical particulate and soluble Cr(VI) salts, respectively. Both compounds induced concentration-dependent cytotoxicity after a 24h exposure in PHBFs. The relative survival was 87, 46, 26 and 2% after exposure to 0.1, 0.5, 1 and 5 microg/cm(2) PbCrO(4), respectively, and 74, 57, 13 and 0% after exposure to 1, 2.5, 5 and 10 microM Na(2)CrO(4), respectively. Similarly, the amount of chromosome damage increased with concentration after 24h exposure to both compounds. Specifically, 0.1, 0.5 and 1 microg/cm(2) PbCrO(4) damaged 15, 34 and 42% of metaphase cells with the total amount of damage reaching 18, 40 and 66 aberrations per 100 metaphases, respectively. PbCrO(4) (5 microg/cm(2)) induced such profound cell cycle delay that no metaphases were found. Na(2)CrO(4) (1 and 2.5 microM) damaged 18 and 33% of metaphase cells with the total amount of damage reaching 19 and 43 aberrations per 100 metaphases, respectively. Na(2)CrO(4) (5 and 10 microM) induced such profound cell cycle delay that no metaphases were found. Overall the data clearly indicate that Cr(VI) compounds are cytotoxic and genotoxic to human lung cells.     

Individual vocal production in a sperm whale (Physeter macrocephalus) social unit

The vocal repertoires of group‐living animals may communicate individual or group identity. Female and juvenile sperm whales live in long‐term social units that can be assigned to vocal clans based on the pattern of clicks in coda vocalizations. An unusual set of circumstances allowed us to record the vocalizations of photo‐identified individuals within a single social unit over a 41 d period. Using click interpulse intervals, we were able to assign codas to individuals and investigate coda production at the individual level within a social unit for the first time. Adult females in the unit vocalized at approximately equal rates. A calf and juvenile, both male, vocalized less often than the adult females. Repertoires were indistinguishable for all unit members apart from a mother and her calf, which possessed significantly different repertoires—even from one another. We suggest that similarity among the coda repertoires of most unit members indicates a function in advertising unit identity. In contrast, the distinctive repertoires of the calf and its mother may facilitate reunions between these whales. We hypothesize that sperm whales may be able to vary their vocal repertoires as their reproductive status alters the trade‐off between the benefits of individual and group identification.     

Characterization of 18 SNP markers for sperm whale (Physeter macrocephalus)

We report the characterization of 18 new single nucleotide polymorphism (SNP) markers for an endangered species, the sperm whale (Physeter macrocephalus), developed using a targeted gene approach. SNP markers were derived from autosomal regions of the genome using primers originally characterized for genome mapping in other mammals. These SNP markers are the first to be designed for genotyping sperm whale populations and will provide a necessary addition to the genetic tools employed for understanding population structure on a global scale and for developing a conservation management strategy for this endangered species.     

Cytotoxicity and genome-wide microarray analysis of intestinal smooth muscle cells in response to hexavalent chromium induction

Chronic ingestion of high concentrations of hexavalent chromium Cr(VI) in drinking water induces intestinal tumors in mice; however, information on its toxicity on intestinal smooth muscle cells is limited. The present study aimed to assess the in vitro and in vivo toxicological effects of Cr(VI) on intestinal smooth muscle cells. Human intestinal smooth muscle cells (HISM cells) were cultured with different concentrations of Cr(VI) to evaluate effects on cell proliferation ability, oxidative stress levels, and antioxidant system. Furthermore, tissue sections in Cr(VI) exposed rabbits were analyzed to evaluate toxicity on intestinal muscle cells in vivo. Gene chips were utilized to assess differential gene expression profiles at the genome-wide level in 1 μmol/L Cr(VI) treated cells. Intestinal tissue biopsy results showed that Cr(VI) increased the incidences of diffuse epithelial hyperplasia in intestinal jejunum but caused no obvious damage to the structure of the muscularis. Cell proliferation analysis revealed that high concentrations (≥64 μmol/L) but not low concentrations of Cr(VI) (≤16 μmol/L) significantly inhibited the growth of HISM cells. For oxidative stress levels, the expression of reactive oxygen species (ROS) and nitric oxide (NO) was elevated at high concentrations (≥64 μmol/L) but not at low concentrations of Cr(VI) (≤16 μmol/L). In addition, dose-dependent increases in the activity of oxidized glutathione (GSSH)/total-glutathione (T-GSH) were also observed. Gene chip screened 491 differentially expressed genes including genes associated with cell apoptosis, oxidations, and cytoskeletons. Some of these differentially expressed genes may be unique to smooth muscle cells in response to Cr(VI) induction.     

Female philopatry in coastal basins and male dispersion across the North Atlantic in a highly mobile marine species, the sperm whale (Physeter macrocephalus)

The mechanisms that determine population structure in highly mobile marine species are poorly understood, but useful towards understanding the evolution of diversity, and essential for effective conservation and management. In this study, we compare putative sperm whale populations located in the Gulf of Mexico, western North Atlantic, Mediterranean Sea and North Sea using mtDNA control region sequence data and 16 polymorphic microsatellite loci. The Gulf of Mexico, western North Atlantic and North Sea populations each possessed similar low levels of haplotype and nucleotide diversity at the mtDNA locus, while the Mediterranean Sea population showed no detectable mtDNA diversity. Mitochondrial DNA results showed significant differentiation between all populations, while microsatellites showed significant differentiation only for comparisons with the Mediterranean Sea, and at a much lower level than seen for mtDNA. Samples from either side of the North Atlantic in coastal waters showed no differentiation for mtDNA, while North Atlantic samples from just outside the Gulf of Mexico (the western North Atlantic sample) were highly differentiated from samples within the Gulf at this locus. Our analyses indicate a previously unknown fidelity of females to coastal basins either side of the North Atlantic, and suggest the movement of males among these populations for breeding.     

Body fat and condition in sperm whales,Physeter macrocephalus,from southern Australian waters

Blubber thickness (n=102) and lipid content (n=37) were measured in sperm whales from three mass stranding events on the west and north-west coasts of Tasmania, Australia in February 1998. Blubber thickness was highly variable, ranging from 43.0 to 168.0 mm (mean 98.4+/-18.4 mm) while lipid fat content, also highly variable, ranged from 16.19 to 89.34% (mean 49.2+/-17.9%). Blubber thickness was significantly and positively related to total length, but a blubber thickness index based on the residuals of this relationship was not related to age, sex or reproductive condition. No relationship was found between blubber thickness index and blubber lipid content, indicating that blubber thickness may not provide a comprehensive indication of body fat condition in sperm whales when only measured at a single site. Blubber lipid content was not related to total length, age or sex. Blubber lipid content was stratified vertically throughout the blubber layer, suggesting that the inner blubber layer may be a more active site for lipid deposition and mobilisation, while the outer blubber layer may serve in a structural or thermoregulatory role. The social structure and foraging ecology of this species may serve to minimise the need to rely on stored energy reserves to meet reproductive energy requirements. In addition, the broader role of blubber for structural, buoyancy and insulative functions coupled with high individual variability may cause a lack of obvious relationships between these variables and body size, age, sex and reproductive state in this species.     

Residency and abundance of sperm whales (Physeter macrocephalus) in Nemuro Strait,Hokkaido, Japan

We examined the trend in residence patterns and abundance of male sperm whales in Nemuro Strait, Japan, based on long-term photo-identification (1,513 survey days, total 2,969 photos) between 2006 and 2017. A total of 225 unique individuals were identified during this study, with an average of 36 (SE = 2.55) new individuals identified in each season. The model chosen by maximum likelihood suggests that residence time around Nemuro Strait is 769 (SE = 372.4) days, with individuals staying in the strait about 48 days (SE = 8.36) per year. While the migration patterns of male sperm whales visiting this area are still unclear, these findings along with previous studies suggest that males move from one breeding area to another neighboring area every several weeks, shifting their home ranges gradually over a period of a few years. The abundance of sperm whales in Nemuro Strait varied greatly from year to year; from 28 (95%CI: 24–44) in 2015 and (95%CI: 22–48) in 2016 to 66 (95%CI: 57–84) in 2011. This study provides important knowledge of abundance and residency for Nemuro Strait, information which will contribute to further research on the social structure and movement pattern of male sperm whales.     

Prolonged exposure to particulate Cr(VI) is cytotoxic and genotoxic to fin whale cells

BackgroundHexavalent chromium [Cr(VI)] is a human lung carcinogen and global marine pollutant. High Cr concentrations, resembling the ones observed in occupationally exposed workers, have been observed in fin whales (Balaenoptera physalus) in the Gulf of Maine. This outcome suggests Cr might be disrupting the health of fin whale populations. Indeed, Cr in acute (24 h) exposure does cause toxicity in fin whale cells. However, human cell culture data indicate prolonged exposures (120 h) induce a higher amount of toxicity compared to 24 h exposure due to an inhibition of homologous recombination repair. However, whether prolonged exposure causes similar outcomes in fin whale cells is unknown.ObjectiveDue to the importance of assessing prolonged exposure toxicity, this study focuses on characterizing acute and prolonged exposure of Cr(VI) in male and female fin whale cells.MethodsCytotoxicity was measured by the clonogenic assay, also known as colony forming assay, which measures the ability of cells to proliferate and form colonies after the treatment. DNA double strand breaks were analyzed by neutral comet assay. Clastogenicity was measured using the chromosome aberration assay. Intracellular Cr levels were measured with Graphite Furnace Atomic Absorption Spectrometry (GFAAS) with Syngistix Software.ResultsIn this study, we demonstrate that particulate Cr(VI) induces cytotoxicity and genotoxicity in a treatment-dependent manner after 24 h and 120 h exposures. Cytotoxicity levels were generally low with relative survival above 64 %. DNA double strand break data and chromosome aberration data were elevated after a 24 h exposure, but decreased after a 120 h exposure. While cytotoxicity was similar after 24 h and 120 h exposures, less DNA double strand breaks and chromosomal instability occurred with prolonged exposure.ConclusionParticulate Cr(VI) is cytotoxic and genotoxic to fin whale cells after acute and prolonged exposures. The reduction of genotoxicity we have observed after 120 h exposure may be partly explained by lower intracellular Cr levels after 120 h. However, the decrease in intracellular levels is not reflected by a similar decrease in chromosome aberrations suggesting other mechanisms may be at play. Male fin whale cells appear to be more susceptible to the genotoxic effects of particulate Cr(VI) while female cells are less susceptible possibly due to increased cell death of damaged cells, but more work is needed to clarify if this outcome reflects a sex difference or interindividual variability. Overall, the study shows particulate Cr(VI) does induce toxicity at both acute and prolonged exposures in fin whales cells indicating Cr(VI) exposure is a health risk for this species.     

Spatio-temporal distribution of sperm whales (Physeter macrocephalus) off Kaikoura,New Zealand,in relation to bathymetric features

Using a shore-based station we monitored the position of sperm whales (Physeter macrocephalus) within the Kaikoura submarine canyon from 2010 to 2012. We tracked sperm whales using a theodolite station for a total of 290 days. We extracted the distance from the nearest coast, the depth and the bathymetric slope using ArcGIS 10.1. We estimated the seasonal spatial distribution of sperm whales using general additive models. The distribution varied significantly between seasons; individuals were found in deeper water and further offshore in the spring than in winter. This study improved our understanding of the variability of sperm whale distribution patterns off Kaikoura. We determined that the distribution was linked to the bathymetric features and we hypothesized that whales adapted their use of the submarine canyon in relation to food aggregation. We would encourage further studies to evaluate the sperm whale relationship with oceanographic variables off Kaikoura.     

Vocal repertoires and insights into social structure of sperm whales (Physeter macrocephalus) in Mauritius,southwestern Indian Ocean

Sperm whales communicate using codas (stereotyped click sequences). Females and juveniles live in long-term social units, and units with similar coda repertoires share vocal clan membership. Vocal clans exhibit culturally defined differences in their multilevel social structure. Here, we aimed to identify different social units among sperm whales in Mauritius in the southwestern Indian Ocean, and to describe and compare their coda repertoires to investigate the presence of different vocal clans. We conducted six boat-based surveys between 2008 and 2013, during which 101 different individuals were photo-identified. Analysis of associations between 22 resighted individuals divided them into four candidate social units, but a lack of resightings impedes solid delineation of social units. Based on number and rhythm of clicks, at least 24 discrete coda types were detected among 4,767 analyzed codas using two different classification methods. Comparison of coda repertoires recorded from seven sperm whale groupings revealed the possible existence of two sympatric vocal clans, but the size differences of recorded repertoires warrant caution of interpretation. To further evaluate social structuring and the presence of different vocal clans in this region, future surveys should aim at obtaining long-term sighting and acoustic data, and cover a more extensive area.     

基于同位素标记相对和绝对定量技术研究耐铬(VI)菌株CM01的蛋白定量组学

【背景】六价铬[Cr(VI)]作为一种高毒性的重金属污染物,用微生物学方法还原Cr(VI)既经济又环保。【目的】探究耐铬菌株CM01中与耐受或还原铬Cr(VI)有关的蛋白或基因,从分子水平上阐述其耐铬Cr(VI)机制。【方法】运用同位素标记相对和绝对定量技术(isobarictagsforrelativeand absolute quantitation techniques,iTRAQ)分析耐铬菌株CM01在有无铬Cr(VI)胁迫下的蛋白表达差异。运用RT-qPCR技术验证4种与代谢途径有关的上调蛋白的m RNA表达量。运用紫外分光光度计测定细胞表面疏水性。【结果】共鉴定到2 570个蛋白,其中存在显著差异的蛋白数为646个,上调蛋白数量343个,下调蛋白数量303个。CM01中的Fe/Mn超氧化物歧化酶、甜菜碱醛脱氢酶、磷酸戊糖途径、磷酸肌醇代谢途径、氨基酸代谢共同参与了菌株对于外源性Cr(VI)的响应机制。RT-qPCR实验结果表明,与代谢途径有关的4种蛋白其基因转录水平和蛋白水平一致。对照组CM01的细胞表面疏水性高于实验组。【结论】初步探索了耐铬菌的Cr(VI)响应机制,为利用微生物治理环境污染提供了理论支持和新的研究思路。     

Effects of taurine on oxidative stress parameters and chromium levels altered by acute hexavalent chromium exposure in Mice Kidney tissue

The kidney has been regarded as a critical organ of toxicity induced by acute exposure to hexavalent chromium [Cr(VI)] compounds. Reactive intermediates and free radicals generated during reduction process might be responsible for Cr(VI) toxicity. In this study, the effects of pretreatment or posttreatment of taurine on Cr(VI)-induced oxidative stress and chromium accumulation in kidney tissue of Swiss albino mice were investigated. Single intraperitoneal (ip) potassium dichromate treatment (20 mgCr/kg), as Cr(VI) compound, significantly elevated the level of lipid peroxidation as compared with the control group (p<0.05). This was accompanied by significant decreases in nonprotein sulfhydryls (NPSH) level, superoxide dismutase (SOD), and catalase (CAT) enzyme activities as well as a significant chromium accumulation (p<0.05). Taurine administration (1 g/kg, ip) before or after Cr(VI) exposure resulted in reduction of lipid peroxidation levels and improvement in SOD enzyme activity (p<0.05). On the other hand, administration of the antioxidant before Cr(VI) exposure restored the NPSH level and CAT enzyme activity and also reduced tissue chromium levels (p<0.05), whereas postreatment had only slight effects on these parameters. In view of the results, taurine seems to exert some beneficial effects against Cr(VI)-induced oxidative stress and chromium accumulation in mice kidney tissue.     

Cephalopod remains from stomachs of sperm whales (Physeter macrocephalus) that mass‐stranded along the Oregon coast

On 16 June 1979, a herd of 41 sperm whales stranded near the mouth of the Siuslaw River in Florence, Oregon. The stomach contents from 32 whales were collected, identified to the lowest taxonomic level possible, enumerated, and measured. A total of 20,247 cephalopod lower beaks that represented 24 species from 14 different families were recovered. The most numerous species were Histioteuthis hoylei (25.9%), Taonius borealis (12.9%), Galiteuthis phyllura (11.2%), Gonatopsis/Berryteuthis type (10.9%), and Moroteuthis robusta (10.7%). Reconstructed estimates of mass indicated that M. robusta contributed almost 50% of the total mass of cephalopods consumed, followed by H. hoylei (19.3%), and T. borealis (7.0%). The most important species in the diet of stranded whales were M. robusta, H. hoylei, T. borealis, G. phyllura, Octopoteuthis deletron, and Gonatopsis/Berryteuthis type. There were significant differences in the diet of males and females, but no differences between sperm whales of different age groups. Overall, sperm whales primarily consumed small cephalopods that were likely eaten south of 45ºN in or near the California Current System. This study provides new estimates of the food habits of sperm whales in the northeast Pacific from one of the largest strandings of this species.     

Population dynamics of sperm whales (Physeter macrocephalus) in Guadeloupe,French Caribbean: A mark-recapture study from 2001 to 2013

Sperm whales are present along the Caribbean islands in family groups of mature females, juveniles, and newborns. Their abundance and demographic rates remain poorly known. Using photo-identification data, we estimated the abundance and annual survival of sperm whales in Guadeloupe from January to April 2001–2013. A total of 1,492 photographs of tail flukes were scored (ranging from 1 to 5) in terms of quality (Q) and marking (M) level. A total of 789 photographs were selected with Q ≥ 3 and M ≥ 3, corresponding to 109 individually identified adult females or immatures of both sexes. Capture histories were built and analyzed using the Pollock's robust design capture-mark-recapture model. The study consisted of 13 primary periods sequenced in 5–8 secondary periods. Mean annual survival rate of residents was 0.945, 95% CI [0.864, 0.979]. Abundance varied between years from 75 to 35 individuals. The geometric growth rate over the period 2001–2013 was 0.938, 95% CI [0.878, 0.997], suggesting a population decline of −6.2%/year. Sperm whales are moving between Guadeloupe and Dominica, with a high temporary emigration rate (0.163) and 50% of the individuals of Guadeloupe resighted in Dominica. These results highlight the fragility of the population and the necessity to consider it as one unit.     

The cytotoxicity and genotoxicity of hexavalent chromium in Steller sea lion lung fibroblasts compared to human lung fibroblasts

In this study we directly compared soluble and particulate chromate cytotoxicity and genotoxicity in human (Homo sapiens) and sea lion (Eumetopias jubatus) lung fibroblasts. Our results show that hexavalent chromium induces increased cell death and chromosome damage in both human and sea lion cells with increasing intracellular chromium ion levels. The data further indicate that both sodium chromate and lead chromate are less cytotoxic and genotoxic to sea lion cells than human cells, based on an administered dose. Differences in chromium ion uptake explained some but not all of the reduced amounts of sodium chromate-induced cell death. By contrast, uptake differences could explain the differences in sodium chromate-induced chromosome damage and particulate chromate-induced toxicity. Altogether they indicate that while hexavalent chromium induces similar toxic effects in sea lion and human cells, there are different mechanisms underlying the toxic outcomes.     

Cytoplasmic and Mitochondrial Creatine Kinases from the Skeletal Muscle of Sperm Whale (Physeter macrocephalus). Molecular Cloning and Enzyme Characterization

We have amplified two cDNAs, coding for creatine kinases (CKs), from the skeletal muscle of sperm whale Physeter macrocephalus by PCR, and cloned these cDNAs into pMAL plasmid. These are the first CK cDNA and deduced amino acid sequences from cetaceans to be reported. One of the two amino acid sequences is a cytoplasmic, muscle-type isoform (MCK), while the other was identified as a sarcomeric, mitochondrial isoform (sMiCK) that included a mitochondrial targeting peptide. The amino acid sequences of sperm whale MCK and sMiCK showed 94–96% sequence identity with corresponding isoforms of mammalian CKs, and all of the key residues necessary for CK function were conserved. The phylogenetic analyses of vertebrate CKs with three independent methods (neighbor-joining, maximum-likelihood and Bayes) supported the clustering of sperm whale MCK with Bos and Sus MCKs, in agreement with the contemporary view that these groups are closely related. Sperm whale MCK and sMiCK were expressed in Escherichia coli as a fusion protein with maltose-binding protein, and the kinetic constants (K _m, K _d and k _cat) were determined for the forward reaction. Comparison of kinetic constants with those of human and mouse CKs indicated that sperm whale MCK has a comparable affinity for creatine (K _m^Cr = 9.38 mM) to that of human MCK, and the sMiCK has two times higher affinity for creatine than the human enzyme. Both the MCK and sMiCK of sperm whale display a synergistic substrate binding (K _d /K _m = 3.1–7.8) like those of other mammalian CKs.     

Histological analysis of the nasal roof cartilage in a neonate sperm whale (Physeter macrocephalus – Mammalia, Odontoceti)

The nasal roof cartilage of a neonate sperm whale (Physeter macrocephalus) was examined by gross dissection and routine histology. This cartilage is part of the embryonic Tectum nasi and is a critical feature in the formation of the massive sperm whale forehead. In neonates as well as in adults, the blade-like nasal roof cartilage extends diagonally through the huge nasal complex from the bony nares to the blowhole on the left side of the rostral apex of the head. It accompanies the left nasal passage along its entire length, which may reach several meters in adult males. The tissue of the nasal roof cartilage in the neonate whale shows an intermediate state of development. For example, in embryos and fetuses, the nasal roof cartilage consists of hyaline cartilage, but in adult sperm whales, it also includes elastic fibers. In our neonate sperm whale, the nasal roof cartilage already consisted of adult-like elastic cartilage. In addition, the active or growing, layer of the perichondrium was relatively thick compared to that of fetuses, and a large number of straight elastic fibers that were arranged perpendicularly to the long axis of the nasal roof cartilage were present. These neonatal features can be interpreted as characteristics of immature and growing cartilaginous tissue. An important function of the nasal roof cartilage may be the stabilization of the left nasal passage, which is embedded within the soft tissue of the nasal complex. The nasal roof cartilage with its elastic fibers may keep the nasal passage open and prevent its collapse from Bernoulli forces during inhalation. Additionally, the intrinsic tension of the massive nasal musculature may be a source of compression on the nasal roof cartilage and could explain its hyaline character in the adult. In our neonate specimen, in contrast, the cartilaginous rostrum (i.e., mesorostral cartilage) consisted of hyaline cartilage with an ample blood supply. The cartilaginous rostrum does not change its histological characteristics during development, but its function in adults is still not understood.     

Effect of incubation temperature after devitrification on quality parameters in human sperm cells

Sperm cryopreservation is common in assisted reproduction laboratories, providing a therapeutic option for several clinical conditions. This process has been optimized; however, the effect of post-thaw incubation temperature has been poorly studied. This work analyzed the effect of incubation temperature after devitrification on human sperm quality. Spermatozoa from normozoospermic donors were cryopreserved by vitrification. After devitrification, the spermatozoa were separated into two aliquots: (i) incubated at room temperature (RT, 22-25 °C) and (ii) incubated at 37 °C. Reactive oxygen species (ROS), viability, mitochondrial membrane potential (ΔΨM), phosphatidylserine externalization and motility were analyzed immediately after devitrification (control) and after 2, 4 and 6 h. Spermatozoa incubated at RT showed a conserved viability and ΔΨM compared to the control, while the incubation at 37 °C promoted a decrease in these parameters. The ROS levels were increased at both incubation conditions. The progressive motility was decreased in all experimental groups and the decrease was more pronounced under incubation at RT. No increase in phosphatidylserine externalization was observed. In conclusion, prior to use in assisted reproduction procedures, devitrified spermatozoa at RT conserve a better viability and ΔΨM than at 37 °C.     

Role of direct reactivity with metals in chemoprotection by N-acetylcysteine against chromium(VI), cadmium(II), and cobalt(II)

The antioxidant N-acetylcysteine (NAC) is widely used for the assessment of the role of reactive oxygen species (ROS) in various biological processes and adverse drug reactions. NAC has been found to effectively inhibit the toxicity of carcinogenic metals, which was attributed to its potent ROS-suppressive properties. However, the absence of redox activity among some metals and findings from genetic models suggested a more diverse, smaller role of oxidative stress in metal toxicity. Here, we examined mechanisms of chemoprotection by NAC against Cd(II), Co(II), and Cr(VI) in human cells. We found that NAC displayed a broad-spectrum chemoprotective activity against all three metals, including suppression of cytotoxicity, apoptosis, p53 activation, and HSP72 and HIF-1α upregulation. Cytoprotection by NAC was independent of cellular glutathione. NAC strongly inhibited the uptake of all three metals in histologically different types of human cells, explaining its high chemoprotective potential. A loss of Cr(VI) accumulation by cells was caused by NAC-mediated extracellular reduction of chromate to membrane-impermeative Cr(III). Suppression of Co(II) uptake resulted from a rapid formation of Co(II)–NAC conjugates that were unable to enter cells. Our results demonstrate that NAC acts through more than one mechanism in preventing metal toxicity and its chemoprotective activity can be completely ROS-independent. Good clinical safety and effectiveness in Co(II) sequestration suggest that NAC could be useful in the prevention of tissue accumulation and toxic effects of Co ions released by cobalt–chromium hip prostheses.