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As an emerging two-dimensional nanomaterial with promising prospects, mono- or few-layer black phosphorus (BP) is potentially toxic to humans. We investigated the effects of two types of BPs on adult male mice through intratracheal instillation. Using the flow cytometry method, the generation, migration, and recruitment of immune cells in different organs have been characterized on days 1, 7, 14, and 21 post-exposure. Compared with small BP (S-BP, lateral size at ∼188 nm), large BP (L-BP, lateral size at ∼326 nm) induced a stronger stress lymphopoiesis and B cell infiltration into the alveolar sac. More importantly, L-BP dramatically increased peripheral neutrophil (NE) counts up to 1.9-fold on day 21 post-exposure. Decreased expression of the CXCR4 on NEs, an important regulator of NE retention in the bone marrow, explained the increased NE release into the circulation induced by L-BP. Therefore, BP triggers systemic inflammation via the disruption of both the generation and migration of inflammatory immune cells.

The coronavirus disease 2019 (COVID-19) has swept the world and still afflicts humans. As an effective means of protection, wearing masks has been widely adopted by the general public. The massive use of disposable masks has raised some emerging environmental and bio-safety concerns: improper handling of used masks may transfer the attached pathogens to environmental media; disposable masks mainly consist of polypropylene (PP) fibers which may aggravate the global plastic pollution; and the risks of long-term wearing of masks are elusive. To maximize the utilization and minimize the risks, efforts have been made to improve the performance of masks (

Black phosphorus (BP) has extensive applications in various fields. The release of BP into aquatic ecosystems and the potential toxic effects on aquatic organisms are becoming major concerns. Here, we investigated the developmental toxicity of few-layered BP toward the zebrafish. We found that BP could adsorb on the surface of the chorion and could subsequently penetrate within the embryo. After exposure of embryos to 10 mg/L BP, developmental malformations appeared at 96 hpf, especially heart deformities such as pericardial edema and bradycardia, accompanied by severe circulatory system failure. Using transgenic zebrafish larvae, we further characterized cardiovascular defects with cardiac enlargement and impaired cardiac vessels as indicators of damage to the cardiovascular system upon BP exposure. We performed transcriptomic analysis on zebrafish embryos treated with a lower concentration of 2 mg/L. The results showed disruption in genes associated with muscle development, oxygen involved processes, focal adhesion, and VEGF and MAPK signaling pathways. These alterations also indicated that BP carries a risk of developmental perturbation at lower concentrations. This study provides new insights into the effects of BP on aquatic organisms.

Tetrabromobisphenol A (TBBPA), a ubiquitous environmental pollutant, has been implicated in developmental toxicity of aquatic animals. However, the impact of TBBPA on development and the related mechanism have not been fully elucidated. In this study, using a live imaging technique and transgenic labeling of zebrafish embryos, we described the toxic effects of TBBPA on hematopoietic development in zebrafish. We demonstrated that TBBPA induced erythroid precursor expansion in the intermediate cell mass (ICM), which perturbed the onset of blood circulation at 24-26 hours postfertilization (hpf). Consequently, excessive blood cells accumulated in the posterior blood island (PBI) and vascular cells formed defective caudal veins (CVs), preventing blood cell flow to the heart at 32-34 hpf. We found that the one-cell to 50% epiboly stage was the most sensitive period to TBBPA exposure during hematopoietic development. Furthermore, our results demonstrated that PBI malformation induced by TBBPA resulted from effects on erythroid precursor cells, which might involve THR signaling in complex ways. These findings will improve the understanding of TBBPA-induced developmental toxicity in teleost.

Rare earth element nanoparticles (REE NPs) or agents have been used extensively in various fields. Human exposure to REE NPs is an increasing concern. To date, REE NP-mediated comprehensive immune responses after incorporation into the body remain unclear. In our study, using gadolinium oxide NPs (Gd

The use of commercial products containing engineered nanomaterials in realistic scenarios may lead to the accumulation of exogenous particles in brain tissues. In this study, we simulated the use of silver (Ag) nasal spray in humans using Sprague-Dawley rats at 0.04 mg/kg/day. Silver-containing particles were explicitly identified in the rat brain after the administration of nasal sprays containing colloidal Ag or silver ions (Ag

The widespread application of silver (Ag) nanomaterials raises health concerns due to the adverse effects that can be associated with silver nanoparticles (AgNPs) exposure. AgNPs can be introduced into human bodies via inhalation, either intentionally (intranasal administration of AgNPs) or unintentionally (environmental pollution, accidental release, or occupational exposure). Recent studies have shown that intranasal exposure of experimental animals to AgNPs can lead to the accumulation of Ag in brain tissues. However, there is little information available concerning what forms of Ag (particulate and ionic) exist in brain tissues. This study aimed to investigate whether particulate Ag exists in rat brains following intranasal exposure of AgNPs at 1 mg/kg/day using multiple analytical techniques. The results demonstrated that Ag-containing particles were presented in AgNPs-exposed rat brains, accounting for 20.2–68.1% of the total Ag. The mass concentrations of Ag-containing particles in brain tissues increased with exposure time but only decreased by 37.5% after elimination for 4 weeks upon exposure cessation. The size of Ag-containing particles identified in rat brains was larger than the original AgNPs. The Ag-containing particles identified in the rat brain were composed of multiple elements, including Ag, sulfur, and selenium with atomic percentages of 45.8%, 37.5%, and 16.7%, respectively. The finding highlighted the occurrence and accumulation of transformed AgNPs containing S and Se in rat brains after intranasal exposure to AgNPs, implying potential risks for brain health.

As typical brominated flame retardants (BFRs), tetrabromobisphenol A (TBBPA) and its derivative TBBPA-bis(2,3-dibromopropyl ether) (TBBPA-BDBPE) are ubiquitous in various environmental compartments. However, the potential health risk posed by these compounds, especially at environmentally relevant levels, remains unclear. In this study, using adult male mice, we investigated the toxicity of orally administered TBBPA and TBBPA-BDBPE at an environmentally relevant dose (57 nmol/kg body weight). After a single exposure and daily exposure, we assessed lipid metabolism homeostasis, the transcriptome, and immune cell components in the liver. We found that the single exposure to TBBPA or TBBPA-BDBPE alone increased the number of hepatic macrophages, induced alterations in the levels of lipids, including triacylglycerol and free fatty acids, and caused transcriptome perturbation. The results from the daily administration groups showed that TBBPA and TBBPA-BDBPE both significantly increased the triacylglycerol content; however, the elevation of hepatic macrophages was observed only in the TBBPA-BDBPE treatment group. This study confirmed that environmentally relevant levels of TBBPA and TBBPA-BDBPE are toxic to the liver. Our findings revealed that dysfunction of the liver is a health concern, following exposure to BFRs, even at very low concentrations. The chronic effects induced by TBBPA and its derivatives should be further investigated.

Use of legacy brominated flame retardants (BFRs), including polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecane (HBCD), has been reduced due to adverse effects of these chemicals. Several novel brominated flame retardants (NBFRs), such decabromodiphenyl ethane (DBDPE) and bis(2,4,6-tribromophenoxy) ethane (BTBPE), have been developed as replacements for PBDEs. NBFRs are used in various industrial and consumer products, which leads to their ubiquitous occurrence in the environment. This article reviews occurrence and fate of a select group of NBFRs in the environment, as well as their human exposure and toxicity. Occurrence of NBFRs in both abiotic, including air, water, dust, soil, sediment and sludge, and biotic matrices, including bird, fish, and human serum, have been documented. Evidence regarding the degradation, including photodegradation, thermal degradation and biodegradation, and bioaccumulation and biomagnification of NBFRs is summarized. The toxicity data of NBFRs show that several NBFRs can cause adverse effects through different modes of action, such as hormone disruption, endocrine disruption, genotoxicity, and behavioral modification. The primary ecological risk assessment shows that most NBFRs exert no significant environmental risk, but it is worth noting that the result should be carefully used owing to the limited toxicity data.

Human uptake abundance of microplastics via various pathways, and they accumulate in human liver, kidney, gut and even placenta (especially with a diameter of 1 μm or less). Recent scientific studies have found that exposure to microplastics causes intestinal inflammation and liver metabolic disorder, but it remains largely unknown that whether the damage and inflammation may cause further development of severe diseases. In this study, we discovered one of such potential diseases that may be induced by the exposure to small-sized microplastics (with a diameter of 1 μm) performing a multi-organ and multi-omics study comprising metabolomics and microbiome approaches. Unlike other animal experiments, the dosing strategy was applied in mice according to the daily exposure of the highly exposed population, which was more environmentally relevant and reflective of real-world human exposure. Our studies on the gut-liver axis metabolism have shown that the crosstalk between the gut and liver ultimately leaded to insulin resistance and even diabetes. We proactively verified this hypothesis by measuring the levels of fasting blood glucose and fasting insulin, which were found significantly elevated in the mice with microplastics exposure. These results indicate the urgent need of large-scale cohort evaluation on epidemiology and prognosis of insulin resistance after microplastics exposure in future.

Although the abiotic and biotic transformation/degradation (T/D) processes of tetrabromobisphenol A (TBBPA) have been widely investigated in model experiments, few reviews have focused on these processes along with their metabolites or degradation products. In this paper, we summarize the current knowledge on the T/D of TBBPA and its derivatives, including abiotic and biotic T/D strategies/conditions, mechanisms, metabolites and environmental occurrences. Various treatments, such as pyrolysis, photolysis, chemical reactions and biotransformation, have been employed to study the metabolic mechanism of TBBPA and its derivatives and to remediate associated contaminated environments. To date, more than 100 degradation products and metabolites have been identified, dominated by less brominated compounds such as bisphenol A, 2,6-dibromo-4-isopropylphenol, 2,6-dibromo-4-hydroxyl-phenol, 2,6-dibromophenol, isopropylene-2,6-dibromophenol, 4-(2-hydroxyisopropyl)-2,6-dibromophenol, etc. It can be concluded that the T/D of TBBPA mainly takes place through debromination and β-scission. In some environmental media and human and animal tissues, brominated metabolites, glucoside and sulfate derivatives are also important T/D products. Here, the T/D products of TBBPA and its derivatives have been most comprehensively presented from the literature in recent 20 years. This review will enhance the understanding of the environmental behaviors of TBBPA-associated brominated flame retardants along with their ecological and health risks.

Gold nanoparticles (AuNPs) are increasingly being used as diagnostic and therapeutic agents owing to their excellent properties; however, there is not much data available on their dynamics in vivo on a single particle basis in a single mouse. Here, we developed a method for the direct analysis of nanoparticles in trace blood samples based on single particle inductively coupled plasma-mass spectrometry (spICP-MS). A flexible, highly configurable, and precisely controlled sample introduction system was designed by assembling an ultralow-volume autosampler (flow rate in the range of 5-5000 μL/min) and a customized cyclonic spray chamber (transfer efficiency up to 99%). Upon systematic optimization, the detection limit of the nanoparticle size (LODsize) of AuNPs in ultrapure water was 19 nm, and the detection limit of the nanoparticle number concentration (LODNP) was 8 × 104 particle/L. Using a retro-orbital blood sampling method and subsequent dilution, the system was successfully applied to track the dynamic changes in size and concentration for AuNPs in the blood of a single mouse, and the recovery for the blood sample was 111.74%. Furthermore, the concentration of AuNPs in mouse blood reached a peak in a short period of time and, then, gradually decreased. This study provides a promising technique for analyzing and monitoring the size and concentration of nanoparticles in ultralow-volume blood samples with low concentrations, making it a powerful tool for analyzing and understanding the fate of nanoparticles in vivo.

As a novel member of the two-dimensional nanomaterial family, mono- or few-layer black phosphorus (BP) with direct bandgap and high charge carrier mobility is promising in many applications such as microelectronic devices, photoelectronic devices, energy technologies, and catalysis agents. Due to its benign elemental composition (phosphorus), large surface area, electronic/photonic performances, and chemical/biological activities, BP has also demonstrated a great potential in biomedical applications including biosensing, photothermal/photodynamic therapies, controlled drug releases, and antibacterial uses. The nature of the BP-bio interface is comprised of dynamic contacts between nanomaterials (NMs) and biological systems, where BP and the biological system interact. The physicochemical interactions at the nano-bio interface play a critical role in the biological effects of NMs. In this review, we discuss the interface in the context of BP as a nanomaterial and its unique physicochemical properties that may affect its biological effects. Herein, we comprehensively reviewed the recent studies on the interactions between BP and biomolecules, cells, and animals and summarized various cellular responses, inflammatory/immunological effects, as well as other biological outcomes of BP depending on its own physical properties, exposure routes, and biodistribution. In addition, we also discussed the environmental behaviors and potential risks on environmental organisms of BP. Based on accumulating knowledge on the BP-bio interfaces, this review also summarizes various safer-by-design strategies to change the physicochemical properties including chemical stability and nano-bio interactions, which are critical in tuning the biological behaviors of BP. The better understanding of the biological activity of BP at BP-bio interfaces and corresponding methods to overcome the challenges would promote its future exploration in terms of bringing this new nanomaterial to practical applications.

Natural mercury-containing nanoparticles (Hg-NPs) have been found in the environment, but the information for Hg-NPs in organisms was still limited. Clarifying the unique roles of Hg-NPs in organisms is crucial to fully understand the health risks of Hg. Herein, liver and muscle tissues of cetaceans were collected to identify the presence and characteristics of Hg-NPs. We found that methylmercury (MeHg) was the dominant species of Hg in muscles, while inorganic Hg (IHg) accounted for 84.4-99.0% (average 94.0%) of Hg in livers. By using transmission electron microscopy (TEM), size exclusion chromatography coupled with inductively coupled plasma mass spectrometry (SEC-ICPMS) and single particle ICPMS (sp-ICPMS), large amounts (9-161 μg/g) of Hg-NPs in livers and small amounts (0.1-0.4 μg/g) in muscles were observed, indicating that Hg-NPs was an important form of Hg in livers. Both small sized (5-40 nm) and large sized (>100 nm) Hg-NPs were identified, which were mainly complexed with selenium (Se) and sulfur (S) as well as a few cadmium (Cd), lead (Pb) and silver (Ag). This study provided direct evidence of Hg-NPs in marine mammals as well as their chemical form and size distribution, which are helpful for further understanding the biogeochemical cycle and health risk of Hg.

Black phosphorus nanosheets (BPs) have exhibited great potential for various applications including biomedicine, thus their potential side effects and corresponding improvement strategy deserve investigation. Here, in vitro and in vivo biological effects of BPs with and without titanium sulfonate ligand (TiL4) modification are investigated. Comparing to bare BPs, BPs with TiL4 modification (TiL4@BPs) can efficiently escape from macrophages uptake, and reduce the cytotoxicity and proinflammation. When intravenously injected into mice, bare BPs trigger significant inflammatory responses, including an increase in peripheral neutrophils accompanied by elevation of a group of inflammatory cytokines, while TiL4@BPs exhibit no such adverse immune responses. The corresponding mechanisms are also discussed. These findings may not only guide the applications of BPs, but also propose an efficient strategy to further improve the biocompatibility of BPs.

Tetrabromobisphenol-A/S (TBBPA/S) analogs have raised substantial concern because of their adverse effects and potential bioaccumulative properties, such as TBBPA bis(allyl ether) (TBBPA-BAE) and TBBPA bis(2,3-dibromopropyl ether) (TBBPA-BDBPE). In this study, a comprehensive method for simultaneous determination of TBBPA/S and nine novel analogs, including TBBPA-BAE, TBBPA-BDBPE, TBBPS-BDBPE, TBBPA mono(allyl ether) (TBBPA-MAE), TBBPA mono(2-bromoallyl ether) (TBBPA-MBAE), TBBPA mono(2,3-dibromopropyl ether) (TBBPA-MDBPE), TBBPS-MAE, TBBPS-MBAE, and TBBPS-MDBPE in biological samples was developed. The distribution patterns and trophic transfer properties of TBBPA/S and analogs in various biological samples collected from the Chinese Bohai Sea were then studied in detail. For the first time, TBBPA-MBAE and TBBPS-BDBPE were detected in biological samples and TBBPA-MBAE was identified as a byproduct. The concentrations of TBBPA and analogs ranged from ND (not detected or below the method detection limit) to 2782.8 ng/g lipid weight (lw), and for TBBPS and analogs ranged from ND to 927.8 ng/g lw. High detection frequencies (86%) for TBBPA, TBBPS and TBBPA-MAE, TBBPA-MDBPE, TBBPS-MAE, TBBPS-MBAE, and TBBPS-MDBPE were obtained. Meanwhile, TBBPA, TBBPS, and these five analogs displayed trophic dilution tendencies due to significantly negative correlations between trophic levels and lipid-corrected concentrations together with the trophic magnification factors (from 0.31 to 0.55). The results also indicated the novel TBBPA-MAE, TBBPA-MBAE, TBBPA-MDBPE, TBBPS-MAE, TBBPS-MBAE, and TBBPS-MDBPE could be generated not only as byproducts, but also as the probable transformation products of commercial TBBPA/S derivatives.

Environmental distribution and concentration of tetrabromobisphenol A bis- (2-hydroxyethyl) ether (TBBPA-DHEE) and tetrabromobisphenol A mono- (hydroxyethyl) ether (TBBPA-MHEE), are obscure due to the lack of available analytical methods. Here two novel immunoassays were established to systematically investigate their distributions in Taizhou, Eastern China. Five monoclonal antibodies against pollutants were generated with two designed haptens through animal immunization. After matched with different coating antigens/antibodies, ELISAs were established (LOD for TBBPA-DHEE, 0.12 ng/mL, based on OVA-M3/mAb-D4G6; LOD for TBBPA-MHEE, 0.79 ng/mL, based on OVA-M3/mAb-D2G6) and applied for investigation of their occurrences at a typical e-waste recycling area after 2-year samples collection, where the total 33 water, 32 soil and 16 biological samples were collected with the highest concentrations of 3.46 ng/mL, 2.76 ng/g (dry weight, dw) and 5.01 ng/g (dw), respectively. Meanwhile, our study also indicated that at the centralizing e-waste recycling sites the serious pollution for both chemicals still existed despite of various efforts. Besides, obvious improvements were observed at an abandoned e-waste recycling region treated and remedied for many years by the local Chinese government. These findings highlight the importance of policy decisions in treatment of pollutants to reduce organic pollutant-related health risks.

Mercury-binding protein profiles in rat plasma at different levels of mercury exposure in vitro and in vivo were systematically investigated using column gel electrophoresis coupled with inductively coupled plasma-mass spectrometry. The current finding provided various protein candidates that may play critical roles in mercury binding and transporting in plasma.

X-ray induced photodynamic therapy (X-ray-PDT) is a promising approach for synergistic cancer radiotherapy and development of suitable radiosensitizers is highly desired. In this paper, we propose black phosphorus/Bi

The neurotoxic effects of methylmercury (MeHg) have been intensively studied. However, the molecular mechanisms responsible for the neurotoxicity of MeHg are not fully understood. To decipher these mechanisms, proteomic and high-throughput mRNA sequencing (RNA-seq) technique were utilized, comprehensively evaluating the cellular responses of human neuroblastoma SK-N-SH cells to MeHg exposure. Proteomic results revealed that MeHg exposure interfered with RNA splicing via splicesome, along with the known molecular mechanisms of mercury-related neurotoxicity (e.g. oxidative stress, protein folding, immune system processes, and cytoskeletal organization). The effects of MeHg on RNA splicing were further verified using RNA-seq. Compared to control, a total of 658 aberrant RNA alternative splicing (AS) events were observed after MeHg exposure. Proteomics and RNA-seq results also demonstrated that mercury chloride (HgCl2) influenced the expression levels of several RNA splicing related proteins and 676 AS events compared to control. These results suggested that RNA splicing could be a new molecular mechanism involved in MeHg and HgCl2 neurotoxicity.

TET2 is a dioxygenase that catalyses multiple steps of 5-methylcytosine oxidation. Although TET2 mutations frequently occur in various types of haematological malignancies, the mechanism by which they increase risk for these cancers remains poorly understood. Here we show that Tet2−/− mice develop spontaneous myeloid, T- and B-cell malignancies after long latencies. Exome sequencing of Tet2−/− tumours reveals accumulation of numerous mutations, including Apc, Nf1, Flt3, Cbl, Notch1 and Mll2, which are recurrently deleted/mutated in human haematological malignancies. Single-cell-targeted sequencing of wild-type and premalignant Tet2−/− Lin−c-Kit+ cells shows higher mutation frequencies in Tet2−/− cells. We further show that the increased mutational burden is particularly high at genomic sites that gained 5-hydroxymethylcytosine, where TET2 normally binds. Furthermore, TET2-mutated myeloid malignancy patients have significantly more mutational events than patients with wild-type TET2. Thus, Tet2 loss leads to hypermutagenicity in haematopoietic stem/progenitor cells, suggesting a novel TET2 loss-mediated mechanism of haematological malignancy pathogenesis., TET2 catalyses DNA demethylation and is mutated in various blood cancers; in particular Tet2 null mice develop haematological neoplasms. Here the authors show that this effect could be due to the increased frequency of mutation associated with TET2 loss in haematopoietic stem/progenitor cells.

Regarding the nanotoxicity, intense attention has been drawn on the capability of nanomaterials in crossing the biological barriers and their potential developmental toxicity. In the current study, the capability of quantum dots (QDs) in crossing the placental barrier and their potential biological influence on erythroid development were closely investigated. When administrated in mice, QDs tended to accumulate in liver, spleen and kidney, and revealed little capacity to cross the placental barrier. QDs had no effect on the survival of E14.5 fetal liver erythroid cells and erythropoiesis.

Graphene oxide (GO) displays promising properties for biomedical applications including drug delivery and cancer therapeutics. However, GO exposure also raises safety concerns such as potential side effects on health. Here, the biological effects of GO suspended in phosphate buffered saline (PBS) with or without 1% nonionic surfactant Tween 80 were investigated. Based on the ex vivo experiments, Tween 80 significantly affected the interaction between GO and peripheral blood from mice. GO suspension in PBS tended to provoke the aggregation of diluted blood cells, which could be prevented by the addition of Tween 80. After intravenous administration, GO suspension with or without 1% Tween 80 was quickly eliminated by the mononuclear phagocyte system. Nevertheless, GO suspension without Tween 80 showed greater accumulation in lungs than that containing 1% Tween 80. In contrast, less GO was found in livers for GO suspension compared to Tween 80 assisted GO suspension. Organs including hearts, livers, lungs, spleens, kidneys, brains, and testes did not reveal histological alterations. The indexes of peripheral blood showed no change upon GO exposure. Our results together demonstrated that Tween 80 could greatly alter GO's biological performance and determine the pattern of its biodistribution in mice.

Due to its antimicrobial activity, nanosilver (nAg) has become the most widely used nanomaterial. Thus far, the mechanisms responsible for nAg-induced antimicrobial properties and nAg-mediated toxicity to organisms have not been clearly recognized. Silver (Ag) ions certainly play a crucial role, and the form of nanoparticles can change the dissolution rate, bioavailability, biodistribution, and cellular uptake of Ag. However, whether nAg exerts direct "particle-specific" effects has been under debate. Here we demonstrated that nAg exhibited a robust inhibition on RNA polymerase activity and overall RNA transcription through direct Ag binding to RNA polymerase, which is separated from the cytotoxicity pathway induced by Ag ions. nAg treatment in vitro resulted in reduced hemoglobin concentration in erythroid cells; in vivo administration of nAg in mice caused profound reduction of hemoglobin content in embryonic erythrocytes, associated with anemia in the embryos. Embryonic anemia and general proliferation deficit due to the significant inhibition on RNA synthesis, at least partially, accounted for embryonic developmental retardation upon nAg administration. To date, there is no conclusive answer to the sources of nAg-mediated toxicity: Ag ions or "particle-specific" effects, or both. We here demonstrated that both Ag ions and nAg particles simultaneously existed inside cells, demonstrating the "Trojan horse" effects of nAg particles in posing biological impacts on erythroid cells. Moreover, our results suggested that "particle-specific" effects could be the predominant mediator in eliciting biological influences on erythroid cells under relatively low concentrations of nAg exposure. The combined data highlighted the inhibitory effect of nAg on RNA polymerase activity through a direct reciprocal interaction.

Although numerous toxicological studies have been performed on carbon nanotubes (CNTs), a few studies have investigated their secondary and indirect effects beyond the primary target tissues/organs. Here, a cascade of events are investigated: the initiating event and the subsequent key events necessary for the development of phenotypes, namely CNT-induced pro-inflammatory effects on iron homeostasis and red blood cell formation, which are linked to anemia of inflammation (AI). A panel of CNTs are prepared including pristine multiwall CNTs (P-MWCNTs), aminated MWCNTs (MWCNTs-NH2 ), polyethylene glycol MWCNTs (MWCNTs-PEG), polyethyleneimine MWCNTs (MWCNTs-PEI), and carboxylated MWCNTs (MWCNTs-COOH). It has been demonstrated that all CNT materials provoke inflammatory cytokine interleukin-6 (IL-6) production and stimulate hepcidin induction, associated with disordered iron homeostasis, irrespective of exposure routes including intratracheal, intravenous, and intraperitoneal administration. Meanwhile, PEG and COOH modifications can ameliorate the activation of IL-6-hepcidin signaling. Long-term exposure of MWCNTs results in AI and extramedullary erythropoiesis. Thus, an adverse outcome pathway is identified: MWCNT exposure leads to inflammation, hepatic hepcidin induction, and disordered iron metabolism. Together, the combined data depict the hazardous secondary toxicity of CNTs in incurring anemia through inflammatory pathway. This study will also open a new avenue for future investigations on CNT-induced indirect and secondary adverse effects.

Carbon nanotubes (CNTs) have promising applications in a wide range of biomedical fields, including imaging, drug/gene delivery and other therapeutics; however, the biosafety concerns of CNTs should be addressed. To date, many reports have documented the toxicological effects on the cells, tissue or organs that are in direct contact with the tubes; however, there is limited evidence to unravel the secondary toxicity upon CNT treatment. Moreover, more effort is needed to gain a definitive understanding of the adverse outcome pathway (AOP) for CNTs, and a pragmatic framework for risk assessment has not been established yet. In the current study, we aimed to decipher the secondary toxicity to joints under CNT exposure. We demonstrated that carboxylated multi-wall CNTs (MWCNTs-COOH) significantly provoked systemic pro-inflammatory responses, leading to synovial inflammation within knee joints, as evidenced by the infiltration of pro-inflammatory cells in the synovium and meniscus. Mechanistic studies showed that MWCNTs-COOH stimulated pro-inflammatory effects by activating macrophages, and the secreted pro-inflammatory cytokines primed the synoviocytes and chondrocytes, resulting in enhanced production of a large array of enzymes involved in articular cartilage degeneration, including matrix metalloproteinase (MMP) members and cyclooxygenase (COX) members, and increased enzymatic activity of MMPs was demonstrated. Blockade of the cytokines by antibodies significantly attenuated the production of these enzymes. Our current study thus suggests that there is a novel secondary toxicity of CNTs, namely a new AOP to understand the indirect effects of carbon nanotubes: synovial inflammation due to the alteration of the priming state of synoviocytes and chondrocytes under CNT-induced systemic inflammatory conditions.

Graphene has promising applications in food packaging, water purification, and detective sensors for contamination monitoring. However, the biological effects of graphene are not fully understood. It is necessary to clarify the potential risks of graphene exposure to humans through diverse routes, such as foods. In the present study, graphene, as the model nanomaterial, was used to test its potential effects on the cell proliferation based on multiple representative cell lines, including HepG2, A549, MCF-7, and HeLa cells. Graphene was characterized by Raman spectroscopy, particle size analysis, atomic force microscopy, and transmission electron microscopy. The cellular responses to graphene exposure were evaluated using flow cytometry, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, and alamarBlue assays. Rat cerebral astrocyte cultures, as the non-cancer cells, were used to assess the potential cytotoxicity of graphene as well. The results showed that graphene stimulation enhanced cell proliferation in all tested cell cultures and the highest elevation in cell growth was up to 60%. A western blot assay showed that the expression of epidermal growth factor (EGF) was upregulated upon graphene treatment. The phosphorylation of EGF receptor (EGFR) and the downstream proteins, ShC and extracellular regulating kinase (ERK), were remarkably induced, indicating that the activation of the mitogen-activated protein kinase (MAPK)/ERK signaling pathway was triggered. The activation of PI3 kinase p85 and AKT showed that the PI3K/AKT signaling pathway was also involved in graphene-induced cell proliferation, causing the increase of cell ratios in the G2/M phase. No influences on cell apoptosis were observed in graphene-treated cells when compared to the negative controls, proving the low cytotoxicity of this emerging nanomaterial. The findings in this study revealed the potential cellular biological effect of graphene, which may give useful hints on its biosafety evaluation and the further exploration of the bioapplication.

A novel brominated flame retardant (BFR), tris(2,3-dibromopropyl) isocyanurate (TBC), as well as hexabromocyclododecanes (HBCDs) and polybrominated diphenyl ethers (PBDEs), were analyzed in 11 species of mollusks collected from nine coastal cities around the Chinese Bohai Sea in 2009 and 2010. The detection frequencies were 100%, 99%, and 77% for PBDEs, HBCDs, and TBC, respectively. Concentrations of ∑HBCDs ranged from below detection limit (nd) to 28.8 ng g(-1) on a dry weight (dw) basis, followed by ∑(12)PBDE (0.01-20.4 ng g(-1) dw) and TBC (nd-12.1 ng g(-1) dw). Statistically significant linear correlations were found among the three BFRs. Positive correlations were found between BFRs concentrations and lipid content in mollusks. The concentrations tend to decrease with increasing trophic levels of the mollusks, implying trophic dilution rather than biomagnifications of the BFRs in the aquatic food chains of the sampling area. Among the 11 mollusks species, Mytilus edulis showed higher bioaccumulation capability than others and was therefore considered to be an appropriate bioindicator of contamination by the BFRs in the Chinese Bohai Sea, in agreement with its previous selection for the biomonitoring of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs). A dramatic decrease in PBDE concentrations in mollusks of the area was found for the time period from 2003 to 2010, with a half-life of only 2.3 ± 1.7 years, reflecting a rapid response of mollusks to the change in pollution of the marine environment.

Brominated flame retardants (BFRs) have been widely used as additives in products to reduce their flammability. Recent findings suggested that some BFRs exhibit neurotoxicity and thus might pose a threat to human health. In this work, a neurotoxicity assay-directed analysis was developed, combining sample cleanup, fractionation, chemical identification, and bioassay. Viability of primary cultured cerebellar granule neurons (CGNs) was used to evaluate the neurotoxicity of extracts or separated fractions from environmental samples. Tetrabromobisphenol A diallyl ether (TBBPA DAE) was identified as the causative toxicant in sediment samples collected from a river near a brominated flame retardant (BFR) manufacturing plant in South China. Liquid chromatography atmospheric pressure chemical ionization tandem mass spectrometry (LC-APCI-MS/MS) was optimized to determine TBBPA DAE levels in the potent fractions and to confirm TBBPA DAE as the key neurotoxicant. On the basis of comparison with the structure of other TBBPA derivatives, the 1-propenyl group in TBBPA DAE appears to be the cause for the neurotoxic potency. The levels of TBBPA DAE in samples along the river were found at up to 49 ng/L for river water, 10,183 ng/g dry weight (dw) in surface sediments, and 42 ng/g dw in soils. According to the distribution of TBBPA DAE in the environmental samples, the manufacturing plant was identified as the release source of TBBPA DAE. To our knowledge, this study is the first to demonstrate potential neurotoxicity induced by TBBPA DAE in real environmental samples.

Tetrabromobisphenol A (TBBPA) can cause diverse adverse effects including neurotoxicity. Emerging TBBPA derivatives, with high structure similarity to the parent compound, are now being concerned. In this study, the potential neurotoxicities of four TBBPA derivatives and their parent compound were studied by cell viability inhibition in rat pheochromocytoma cells (PC12) and the corresponding molecular mechanisms were investigated. The cellular toxicity was correlated with the chemical hydrophobicity. Tetrabromobisphenol A bis(2-hydroxyethyl ether) (TBBPA-BHEE) exhibited the highest cellular toxicity to PC12 due to its lowest hydrophobicity among these 5 tested compounds. Further experiments showed that TBBPA-BHEE disturbed dopamine (DA) secretion and altered acetylcholinesterase (AChE) enzymatic activity in PC12 cells. The molecular mechanism study indicated that TBBPA-BHEE induced cellular toxicity to PC12 cells through ROS-mediated caspase activation to a large extent, which was partially attenuated by the anti-oxidation of Vitamin E. Moreover, in contrast to TBBPA, the occurrence of TBBPA-BHEE toxicity to PC12 was not attributed to activation of mitogen-activated protein kinases (MAPKs) or thyroid hormone (TH) signaling pathway. These findings suggest TBBPA derivatives, especially TBBPA-BHEE, as potential neurotoxins need urgent attention.

Identification of novel brominated contaminants in the environment, especially the derivatives and byproducts of brominated flame retardants (BFRs), has become a wide concern because of their adverse effects on human health. Herein, we qualitatively and quantitatively identified three byproducts of tetrabromobisphenol-S bis(2,3-dibromopropyl ether) (TBBPS-BDBPE), including TBBPS mono(allyl ether) (TBBPS-MAE), TBBPS mono(2-bromoallyl ether) (TBBPS-MBAE) and TBBPS mono(2,3-dibromopropyl ether) (TBBPS-MDBPE) as novel brominated contaminants. Meanwhile, the mass spectra and analytical method for determination of TBBPS-BDBPE byproducts were presented for the first time. The detectable concentrations (dry weight) of TBBPS-MAE, TBBPS-MBAE and TBBPS-MDBPE were in the ranges 28–394 μg/g in technical TBBPS-BDBPE and 0.1–4.1 ng/g in mollusks collected from the Chinese Bohai Sea. The detection frequencies in mollusk samples were 5%, 39%, 95% for TBBPS-MAE, TBBPS-MBAE and TBBPS-MDBPE, respectively, indicating their prevailing in the environment. The results showed that they could be co-produced and leaked into the environment with production process and might be more bioaccumulative and toxic than TBBPS-BDBPE. Therefore, the production and use of TBBPS derivatives lead to unexpected contamination to the surrounding environment. This study also provided an effective approach for identification of novel contaminants in the environment with synthesized standards and Orbitrap high resolution mass spectrometry.

Osteoporosis is one of the leading disorders among aged people. Bone loss results from a number of physiological alterations, such as estrogen decline and aging. Meanwhile, iron overload has been recognized as a risk factor for bone loss. Systemic iron homeostasis is fundamentally governed by the hepcidin-ferroportin regulatory axis, where hepcidin is the key regulator. Hepcidin deficiency could induce a few disorders, of which iron overload is the most representative phenotype. However, there was little investigation of the effects of hepcidin deficiency on bone metabolism. To this end, hepcidin-deficient (Hamp1(-/-)) mice were employed to address this issue. Our results revealed that significant iron overload was induced in Hamp1(-/-) mice. Importantly, significant decreases of maximal loading and maximal bending stress were found in Hamp1(-/-) mice relative to wildtype (WT) mice. Moreover, the levels of the C-telopeptide of type I collagen (CTX-1) increased in Hamp1(-/-) mice. Therefore, hepcidin deficiency resulted in a marked reduction of bone load-bearing capacity likely through enhancing bone resorption, suggesting a direct correlation between hepcidin deficiency and bone loss Targeting hepcidin or the pathway it modulates may thus represent a therapeutic for osteopenia or osteoporosis. (C) 2014 Elsevier B.V. All rights reserved.

Graphene and graphene-based nanomaterials display novel and beneficial chemical, electrical, mechanical, and optical characteristics, which endow these nanomaterials with promising applications in a wide spectrum of areas such as electronics and biomedicine. However, its toxicity on health remains unknown and is of great concern. In the present study, we demonstrated that graphene oxide (GO) induced necrotic cell death to macrophages. This toxicity is mediated by activation of toll-like receptor 4 (TLR4) signaling and subsequently in part via autocrine TNF-α production. Inhibition of TLR4 signaling with a selective inhibitor prevented cell death nearly completely. Furthermore, TLR4-deficient bone marrow-derived macrophages were resistant to GO-triggered necrosis. Similarly, GO did not induce necrosis of HEK293T/TLR4-null cells. Macrophagic cell death upon GO treatment was partially attributed to RIP1-RIP3 complex-mediated programmed necrosis downstream of TNF-α induction. Additionally, upon uptake into macrophages, GO accumulated primarily in cytoplasm causing dramatic morphologic alterations and a significant reduction of the macrophagic ability in phagocytosis. However, macrophagic uptake of GO may not be required for induction of necrosis. GO exposure also caused a large increase of intracellular reactive oxygen species (ROS), which contributed to the cause of cell death. The combined data reveal that interaction of GO with TLR4 is the predominant molecular mechanism underlying GO-induced macrophagic necrosis; also, cytoskeletal damage and oxidative stress contribute to decreased viability and function of macrophages upon GO treatment.

Tetrabromobisphenol A (TBBPA) is one of the most widely used brominated flame retardants (BFRs) and has been frequently detected in the environment and biota. Recent studies have found that derivatives of TBBPA, such as TBBPA bis(allyl) ether (TBBPA BAE) and TBBPA bis(2,3-dibromopropyl) ether (TBBPA BDBPE) are present in various environmental compartments. In this work, using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS), TBBPA allyl ether (TBBPA AE) and TBBPA 2,3-dibromopropyl ether (TBBPA DBPE) were identified in environmental samples and further confirmed by synthesized standards. Soil, sediment, rice hull, and earthworm samples collected near a BFR manufacturing plant were found to contain these two compounds. In sediments, the concentrations of TBBPA AE and TBBPA DBPE ranged from 1.0 to 346.6 ng/g of dry weight (dw) and from 0.7 to 292.7 ng/g of dw, respectively. TBBPA AE and TBBPA DBPE in earthworm and rice hull samples were similar to soil samples, which ranged from below the method limit of detection (LOD

Nanosilver (nAg) is a considerably important nanomaterial due to its unique physical and chemical features and its intrinsic antimicrobial properties. Thus far, nAg has been widely applied in a variety of fields including biomedicine. Considering the safety and adverse influence, investigations into the biological fate and potential toxicity of nAg are essential for its safe and appropriate applications. In the current study, we exposed nAg to BALB/c mice at various concentrations via intraperitoneal (IP) and intravenous (IV) routes. The results showed that nAg was predominantly localized in liver and spleen in mice for both administration methods. Compared to IP administration, nAg was quickly removed and excreted from body with IV administration. The accumulation of nAg in livers caused remarkable hepatic toxicity. For the first time, we demonstrated that nAg had the ability to cross the placental barrier and accumulate in fetuses. Furthermore, the results of nAg tissue distribution in male mice revealed that nAg could pass through the blood-testis barrier, resulting in localization in testis. Additionally, the pharmacokinetic process of nAg in mice was also assessed in this study. Our findings indicated that nAg retention in mouse body could last for more than 4 months, and the silver content in the major recipient organs decreased in a time-dependent manner. Taken together, these results would be of great assistance in clarifying the safety issue of nAg as drug delivery and therapeutic agent, and in the understanding of the mechanisms underlying nAg-meditated toxicity.

The impact of silver nanoparticles (AgNPs) on the central nervous system is a topic with mounting interest and concern and the facts remain elusive. In the current study, the neurotoxicity of commercial AgNPs to rat cerebellum granule cells (CGCs) and the corresponding molecular mechanism are closely investigated. It is demonstrated that AgNPs induce significant cellular toxicity to CGCs in a dose-dependent manner without damaging the cell membrane. Flow cytometry analysis with the Annexin V/propidium iodide (PI) staining indicates that the apoptotic proportion of CGCs upon treatment with AgNPs is greatly increased compared to the negative control. Moreover, the activity of caspase-3 is largely elevated in AgNP-treated cells compared to the negative control. AgNPs are demonstrated to induce oxidative stress, reflected by the massive generation of reactive oxygen species (ROS), the depletion of antioxidant glutathione (GSH), and the increase of intracellular calcium. Histological examination suggests that AgNPs provoke destruction of the cerebellum granular layer in rats with concomitant activation of caspase-3, in parallel to the neurotoxicity of AgNPs observed in vitro. Taken together, it is demonstrated for the first time that AgNPs substantially impair the survival of primary neuronal cells through apoptosis coupled to oxidative stress, depending on the caspase activation-mediated signaling.

Iron is essential for the human being, involving in oxygen transport, energy metabolism and DNA synthesis. Iron homeostasis is tightly governed by the hepcidin-ferroportin axis, of which hepcidin is the master regulator. Excess iron is associated with various diseases including osteopenia and osteoporosis, which are closely related to the alternation of the endogenous estrogen level. To verify the biological effect of estrogen on iron metabolism, we established a mouse model of estrogen deficiency by ovariectomy. We demonstrated that the hemoglobin content and serum iron level decreased, whereas the tissue iron level in liver and spleen increased in the ovariectomized mice. Moreover, the transcription of hepatic hepcidin was elevated in ovariectomized mice compared to the control mice. We further demonstrated that there was an estrogen response element (ERE) in the promoter region of the hepcidin gene. The assay using the luciferase reporter system confirmed the existence of a functional ERE in the hepcidin promoter, as the estradiol treatment reduced hepcidin expression in cells transfected with ERE-intact construct, with no response to estradiol in cells transfected with ERE-devoid construct. In conclusion, estrogen greatly contributes to iron homeostasis by regulating hepatic hepcidin expression directly through a functional ERE in the promoter region of hepcidin gene. These findings might help build a better understanding towards the etiology of postmenopausal osteoporosis accompanied by excess tissue iron (such as iron retention of osteoclasts in bone) under estrogen deficiency.

Quantum dots (QDs) are fluorescent semiconductor nanoparticles that have broad excitation spectra, narrow emission peaks, long fluorescence lifetimes, and the ability to easily conjugate with bio-molecules. Due to these distinct characteristics, QDs represent promising substances in biological imaging and labelling. However, the side and adverse effects of QDs are also widely studied. Herein, we recognize macrophages as the pivotal cells in ingesting QDs, and that the accumulation of QDs inside macrophages leads to significant morphological alterations and a remarkable reduction of their ability to erythrophagocytize in vitro. In a mouse model with chronic exposure to QDs, red blood cell (RBC) retention in spleens and severe splenomegaly were observed, presumably due to attenuated macrophagic erythrophagocytosis in vivo. Importantly, we demonstrated that QDs greatly inhibited the Rho-associated kinase (ROCK) activity, resulting in impaired fidelity of the actin cytoskeleton and actin-rich structure (such as surface protrusions), which was assumed to be the molecular basis underlying the blunted macrophagic morphology and reduced ability to phagocytize. The combined data provide insights into QDs' intracellular trafficking, localization and biological fate in macrophages, and the resultant impairment to cytoskeleton coupled with inhibition on the ROCK signalling would decrease the macrophagic ability to erythrophagocytize with diminished RBC recycling and splenic RBC retention in animals.

A reliable analytical method was developed here for the simultaneous separation, identification and quantification of ten polybrominated diphenyl ethers (PBDEs), nine methoxylated PBDEs (MeO-PBDEs) and ten hydroxylated PBDEs (OH-PBDEs) in various environmental matrices, including water, soil, sediment, plant, mollusk and fish. PBDEs and MeO-PBDEs were measured by gas chromatography coupled with mass spectrometry (GC/MS) and liquid chromatography coupled with electrospray ionization (negative)-tandem quadrupole mass spectrometry (LC/ESI(−)-MS/MS) for the separation and determination of OH-PBDEs without prior derivatization. After preliminary sample cleaning using acid silica gel, water-impregnated silica column separation of PBDEs, MeO-PBDEs and OH-PBDEs was proved to be rapid, simple, and efficient. For phenolic analytes, the method detection limits (MDLs) were 3.2–11.6 pg/L in water sample and 2.8–18.4 pg/g dry weight in solid samples. For neutral compounds, MDLs were 48.8–150.3 pg/L in water sample and 46.5–170.8 pg/g dry weight in solid samples. The method was validated using six kinds of environmental samples spiked with all analytes at three concentration levels (0.3 ng, 2 ng and 5 ng, respectively) for recovery (71–113%) and repeatability determination (4–12%RSD).

Hexabromocyclododecanes (HBCDs) and tris (2,3-dibromopropyl) isocyanurate (TBC) are both brominated flame retardants, and have been used throughout the world with subsequent concern for potential environment pollution. Here we describe the development of a method for the simultaneous determination of TBC and HBCD diastereoisomers in environmental matrices by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The method recovery ranged from 81% to 93% and limits of detections (LODs) on column were 4.3 pg, 0.5 pg, 0.4 pg, 0.3 pg for TBC, α-HBCD, β-HBCD, and γ-HBCD, respectively, which showed high sensitivity compared to previous published methods. Compared to electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI) was found to be a more sensitive, effective ionization mode for determination of HBCDs. This novel method was further validated by analyzing TBC and HBCDs in biotic and abiotic samples. The concentration ranges of TBC, α, β, γ-HBCD in sediment samples were 136.1-5884.6 ng/g, 9.0-1079.6 ng/g, 1.0-1161.7 ng/g, and 65.4-3964.2 ng/g, respectively. The concentration range for TBC, α-HBCD, β-HBCD, and γ-HBCD in common carp samples were 51.1-1899 ng/g, 21.6-1338 ng/g,LOD-10.9 ng/g, and 1.4-101 ng/g, respectively.

A brominated flame retardant (BFR), hexabrominated heterocyclic tris-(2,3-dibromopropyl) isocyanurate (TBC), was identified,forthe first time, in the natural environment.The chemical was found in river water (2.33-163 ng/L), surface sediments (85.0 ng/g-6.03 microg/g dry weight (dw)), soils (19.6-672 ng/g dw), earthworm (9.75-78.8 ng/g dw), and carp samples (12.0-646 ng/g dw) from a factory-polluted area in southern China. It was found that TBC can strongly adsorb to organic material in sediment, and a trend of decreasing concentration with distance from the source in soil and earthworm samples, combined with calculated Kow (octanol-water partition coefficient) and Koa (octanol-air partition coefficient), suggests its potential ability to undergo regional transportation through dust deposition. Calculated results showed high Kow (log Kow = 7.37) and bioaccumulation factor (BAF) (log BAF = 4.30) of this BFR and indicate that TBC has semivolatile properties and bioaccumulation characteristic in certain biological species. Quantitative structure property relationships (QSPRs) modeling revealed that TBC has Koa (log Koa = 23.68) and Kaw (air-water partition coefficient) (log Kaw = -16.31) values several orders higher than those of other BFRs. The identification of this chemical additive further reminds us that the production and usage of heterocyclic BFRs may cause potential contamination to the surrounding environment