Cytotoxic Evaluation and Determination of Organic and Inorganic Eluates from Restorative Materials.

Over the last years, diverse commercial resin-based composites have dominated as dental filling materials. The purpose of the present study was to determine organic and inorganic eluates from five restorative materials using GC/MS and ICP-OES and to compare the effect on cell survival of human gingival fibroblasts of a conventional and a bioactive resin. Five commercially available restorative materials were employed for this study: ActivaTM Bioactive Restorative, ENA HRi, Enamel plus HRi Biofunction, Fuji II LC Capsule, and Fuji IX Capsule. Disks that were polymerized with a curing LED light or left to set were immersed in: 1 mL methanol or artificial saliva for GC/MS analysis, 5mL deionized water for ICP-OES, and 5mL of culture medium for cell viability. Cell viability was investigated with a modified staining sulforhodamine B assay.The following organic substances were detected: ACP, BHT, BPA, 1,4-BDDMA, CQ, DBP, DMABEE, HEMA, MCE, MeHQ, MOPA, MS, TMPTMA, and TPSb and the ions silicon, aluminum, calcium, sodium, and barium. Activa Bioactive Restorative was found to be biocompatible. Elution of organic substances depended on material's composition, the nature of the solvent and the storage time. Ions' release depended on material's composition and storage time. The newly introduced bioactive restorative was found to be more biocompatible.

Impacts of foodborne inorganic nanoparticles on the gut microbiota-immune axis: potential consequences for host health.

BACKGROUND: In food toxicology, there is growing interest in studying the impacts of foodborne nanoparticles (NPs, originating from food additives, food supplements or food packaging) on the intestinal microbiome due to the important and complex physiological roles of these microbial communities in host health. Biocidal activities, as described over recent years for most inorganic and metal NPs, could favour chronic changes in the composition and/or metabolic activities of commensal bacteria (namely, intestinal dysbiosis) with consequences on immune functions. Reciprocally, direct interactions of NPs with the immune system (e.g., inflammatory responses, adjuvant or immunosuppressive properties) may in turn have effects on the gut microbiota. Many chronic diseases in humans are associated with alterations along the microbiota-immune system axis, such as inflammatory bowel diseases (IBD) (Crohn's disease and ulcerative colitis), metabolic disorders (e.g., obesity) or colorectal cancer (CRC). This raises the question of whether chronic dietary exposure to inorganic NPs may be viewed as a risk factor facilitating disease onset and/or progression. Deciphering the variety of effects along the microbiota-immune axis may aid the understanding of how daily exposure to inorganic NPs through various foodstuffs may potentially disturb the intricate dialogue between gut commensals and immunity, hence increasing the vulnerability of the host. In animal studies, dose levels and durations of oral treatment are key factors for mimicking exposure conditions to which humans are or may be exposed through the diet on a daily basis, and are needed for hazard identification and risk assessment of foodborne NPs. This review summarizes relevant studies to support the development of predictive toxicological models that account for the gut microbiota-immune axis. CONCLUSIONS: The literature indicates that, in addition to evoking immune dysfunctions in the gut, inorganic NPs exhibit a moderate to extensive impact on intestinal microbiota composition and activity, highlighting a recurrent signature that favours colonization of the intestine by pathobionts at the expense of beneficial bacterial strains, as observed in IBD, CRC and obesity. Considering the long-term exposure via food, the effects of NPs on the gut microbiome should be considered in human health risk assessment, especially when a nanomaterial exhibits antimicrobial properties.

Can inorganic elements affect herpesvirus infections in European eels?

In combination, pollution and pathogens represent a serious threat to the health of European eels that has been increasingly recognized. Thus, the impact of contaminants, cadmium, lead, mercury, and selenium, on anguillid herpesvirus 1 infection in wild European eels has been evaluated. Despite the small sample size, results indicate that selenium and mercury concentrations may compromise the European eel immune system as herpesvirus infection was more prevalent in specimens with higher Hg and Se hepatic concentrations.

Cytotoxicity of organic and inorganic compounds to primary cell cultures established from internal tissues of Chelonia mydas.

Chemical contaminants have been found in the tissues of sea turtles from all over the world; however, very little is known about the effects. Recently, in vitro alternatives to live animal testing have been applied to sea turtles due to their ethical and practical benefits. While primary skin fibroblasts have been established for several species of sea turtle, cells from internal organs are lacking, though they may be more relevant due to the well documented accumulation of contaminants within internal tissues. This study established primary cell cultures from the small intestine, heart, liver, ovary and skin of green turtles (Chelonia mydas). Cells were exposed to ten contaminants typically found in sea turtles to examine potential variations in sensitivity among cells established from different organs. Differences between cells established from different animals were also examined, including a comparison of cells established from a turtle with fibropapillomatosis (FP) and healthy turtles. Loggerhead (Caretta caretta) primary skin cells were also included for species comparisons. Significant differences were found between the organ types, with liver and heart being the least sensitive, and skin being the most sensitive. Overall, variation between the organ types was low. Primary skin fibroblasts may be a suitable and representative cell type for in vitro turtle toxicology research, as it is relatively easy to obtain from healthy live animals. Skin cultures provide a more sensitive indication of effect, and could be used as an early warning of the potential effects of chemical contamination. Some species differences were found but no differences were found between cell cultures from an FP turtle and healthy turtles. When EC50 values were compared to accumulation values from the literature, inorganic contaminants, such as Zn, Cd, Cr, Hg, and Cu were identified as posing a potential risk to sea turtle populations around the world.

Method to characterize inorganic particulates in lung tissue biopsies using field emission scanning electron microscopy.

Humans accumulate large numbers of inorganic particles in their lungs over a lifetime. Whether this causes or contributes to debilitating disease over a normal lifespan depends on the type and concentration of the particles. We developed and tested a protocol for in situ characterization of the types and distribution of inorganic particles in biopsied lung tissue from three human groups using field emission scanning electron microscopy (FE-SEM) combined with energy dispersive spectroscopy (EDS). Many distinct particle types were recognized among the 13 000 particles analyzed. Silica, feldspars, clays, titanium dioxides, iron oxides and phosphates were the most common constituents in all samples. Particles were classified into three general groups: endogenous, which form naturally in the body; exogenic particles, natural earth materials; and anthropogenic particles, attributed to industrial sources. These in situ results were compared with those using conventional sodium hypochlorite tissue digestion and particle filtration. With the exception of clays and phosphates, the relative abundances of most common particle types were similar in both approaches. Nonetheless, the digestion/filtration method was determined to alter the texture and relative abundances of some particle types. SEM/EDS analysis of digestion filters could be automated in contrast to the more time intensive in situ analyses.

Phenotypic responses in Caenorhabditis elegans following chronic low-level exposures to inorganic and organic compounds.

Responses of organisms to sublethal exposure of environmental stressors can be difficult to detect. We investigated phenotypic changes in the tissue of Caenorhabditis elegans via Raman spectroscopy, as well as survival and reproductive output when exposed to chronic low doses of metals (copper, zinc, or silver), an herbicide (diuron), and a pesticide (imidacloprid). Raman spectroscopy measures changes in phenotype by providing information about the molecular composition and relative abundance of biomolecules. Multivariate analysis was used to evaluate the significance of treatment phenotype segregation plots compared with controls. Dose-dependent responses were observed for copper, zinc, silver, and diuron, whereas imidacloprid exposure resulted in a small response over the tested concentrations. Concentration-dependent shifts in nematode biomolecular phenotype were observed for copper. Despite having a dose-dependent reproductive response, silver, diuron, and imidacloprid produced inconsistent biological phenotype patterns. In contrast, there was a clear stepwise change between low concentrations (0.00625-0.5 mg/L) and higher concentration (1-2 mg/L) of ionic zinc. The findings demonstrate that measuring phenotypic responses via Raman spectroscopy can provide insights into the biomolecular mechanisms of toxicity. Despite the lack of consistency between survival and Raman-measured phenotypic changes, the results support the effectiveness of Raman spectroscopy and multivariate analysis to detect sublethal responses of chemicals in whole organisms and to identify toxic effect thresholds. Environ Toxicol Chem 2018;37:920-930. © 2017 SETAC.

Can in vitro assays account for interactions between inorganic co-contaminants observed during in vivo relative bioavailability assessment?

In vitro assays act as surrogate measurements of relative bioavailability (RBA) for inorganic contaminants. The values derived from these assays are routinely used to refine human health risk assessments (HHRA). Extensive in vitro research has been performed on three major inorganic contaminants; As, Cd and Pb. However, the majority of these studies have evaluated the contaminants individually, even in cases when they are found as co-contaminants. Recently, in vivo studies (animal model) have determined that when the three aforementioned contaminants are present in the same soil matrix, they have the ability to influence each other's individual bioavailability. Since in vitro assays are used to inform HHRA, this study investigated whether bioaccessibility methods including the Solubility/Bioavailability Research Consortium (SBRC) assay, and physiologically based extraction test (PBET), have the ability to detect interactions between As, Cd and Pb. Using a similar dosing methodology to recently published in vivo studies, spiked aged (12 years) soil was assessed by evaluating contaminant bioaccessibility individually, in addition to tertiary combinations. In two spiked aged soils (grey and brown chromosols), there was no influence on contaminant bioaccessibility when As, Cd and Pb we present as co-contaminants. However, in a red ferrosol, the presence of As and Pb significantly decreased (p < 0.05) the bioaccessibility of Cd when assessed using gastric and intestinal phases of the SBRC assay and the PBET. Conceivable, differences in key physico-chemical properties (TOC, Fe, Al, P) between the study soils influenced contaminant interactions and bioaccessibility outcomes. Although bioaccessibility methods may not account for interactions between elements as demonstrated in in vivo models, in vitro assessment provides a conservative prediction of contaminant RBA under co-contaminant scenarios.

Searching Online Chemical Data Repositories via the ChemAgora Portal.

ChemAgora, a web application designed and developed in the context of the "Data Infrastructure for Chemical Safety Assessment" (diXa) project, provides search capabilities to chemical data from resources available online, enabling users to cross-reference their search results with both regulatory chemical information and public chemical databases. ChemAgora, through an on-the-fly search, informs whether a chemical is known or not in each of the external data sources and provides clikable links leading to the third-party web site pages containing the information. The original purpose of the ChemAgora application was to correlate studies stored in the diXa data warehouse with available chemical data. Since the end of the diXa project, ChemAgora has evolved into an independent portal, currently accessible directly through the ChemAgora home page, with improved search capabilities of online data sources.

Trophic fate of inorganic and methyl-mercury in a macrophyte-chironomid food chain.

Dietary transfer of mercury (Hg) is central for its effects on higher trophic animals, nonetheless, its driving parameters and characteristics are not well understood. Here we measured Hg species transfer (uptake) from the macrophyte Elodea nuttallii -mimicking tissues incorporation in sediments after decay- to Chironomus riparius. Methyl-Hg (MMHg) was more transferable than inorganic Hg (IHg) from plant's intracellular and cell wall compartments. After 10-d-long exposure, MMHg was predominantly found in MMHg form in the cytosolic compartment (S) of chironomids, while IHg showed similar concentrations in S and insoluble debris (P) compartments. After cessation of Hg species exposure (depuration), only MMHg resulted in a bioaccumulation factor >1. Toxicokinetics modelling indicated a demethylation of MMHg in the S fraction and its concomitant storage in the P fraction as IHg during both uptake and depuration, revealing an elimination and detoxification mechanism. Our data support that MMHg is more transferable than IHg to sensitive subcellular targets as well as bioavailable fraction in chironomids, in line with field studies showing higher MMHg transfer than IHg in food webs. Hence our data point out macrophytes as a potential Hg source to benthic food webs to be considered for enhancing aquatic environment protection during phytoremediation programs.

Inorganic arsenic: A non-genotoxic carcinogen.

Inorganic arsenic induces a variety of toxicities including cancer. The mode of action for cancer and non-cancer effects involves the metabolic generation of trivalent arsenicals and their reaction with sulfhydryl groups within critical proteins in various cell types which leads to the biological response. In epithelial cells, the response is cell death with consequent regenerative proliferation. If this continues for a long period of time, it can result in an increased risk of cancer. Arsenicals do not react with DNA. There is evidence for indirect genotoxicity in various in vitro and in vivo systems, but these involve exposures at cytotoxic concentrations and are not the basis for cancer development. The resulting markers of genotoxicity could readily be due to the cytotoxicity rather than an effect on the DNA itself. Evidence for genotoxicity in humans has involved detection of chromosomal aberrations, sister chromatid exchanges in lymphocytes and micronucleus formation in lymphocytes, buccal mucosal cells, and exfoliated urothelial cells in the urine. Numerous difficulties have been identified in the interpretation of such results, including inadequate assessment of exposure to arsenic, measurement of micronuclei, and potential confounding factors such as tobacco exposure, folate deficiency, and others. Overall, the data strongly supports a non-linear dose response for the effects of inorganic arsenic. In various in vitro and in vivo models and in human epidemiology studies there appears to be a threshold for biological responses, including cancer.

Potential Links between Cytoskeletal Disturbances and Electroneurophysiological Dysfunctions Induced in the Central Nervous System by Inorganic Nanoparticles.

Inorganic nanomaterials have been widely applied in biomedicine. However, several studies have noted that inorganic nanoparticles can enter the brain and induce cytoskeletal remodeling, as well as electrophysiological alterations, which are related to neurodevelopmental disorders and neurodegenerative diseases. The toxic effects of inorganic nanomaterials on the cytoskeleton and electrophysiology are summarized in this review. The relationships between inorganic NPs-induced cytoskeletal and electrophysiological alterations in the central nervous system remain obscure. We propose several potential relationships, including those involving N-methyl-D-aspartate receptor function, ion channels, transient receptor potential channels, and the Rho pathway.

Effects of reduced mitochondrial DNA content on secondary mitochondrial toxicant exposure in Caenorhabditis elegans.

The mitochondrial genome (mtDNA) is intimately linked to cellular and organismal health, as demonstrated by the fact that mutations in and depletion of mtDNA result in severe mitochondrial disease in humans. However, cells contain hundreds to thousands of copies of mtDNA, which provides genetic redundancy, and creates a threshold effect in which a large percentage of mtDNA must be lost prior to clinical pathogenesis. As certain pharmaceuticals and genetic mutations can result in depletion of mtDNA, and as many environmental toxicants target mitochondria, it is important to understand whether reduced mtDNA will sensitize an individual to toxicant exposure. Here, using ethidium bromide (EtBr), which preferentially inhibits mtDNA replication, we reduced mtDNA 35-55% in the in vivo model organism Caenorhabditis elegans. Chronic, lifelong, low-dose EtBr exposure did not disrupt nematode development or lifespan, and induced only mild alterations in mitochondrial respiration, while having no effect on steady-state ATP levels. Next, we exposed nematodes with reduced mtDNA to the known and suspected mitochondrial toxicants aflatoxin B1, arsenite, paraquat, rotenone or ultraviolet C radiation (UVC). EtBr pre-exposure resulted in mild sensitization of nematodes to UVC and arsenite, had no effect on AfB1 and paraquat, and provided some protection from rotenone toxicity. These mixed results provide a first line of evidence suggesting that reduced mtDNA content may sensitize an individual to certain environmental exposures.

Distribution of selenium in zebrafish larvae after exposure to organic and inorganic selenium forms.

Selenium is an essential micronutrient for many organisms, and in vertebrates has a variety of roles associated with protection from reactive oxygen species. Over the past two decades there have been conflicting reports upon human health benefits and detriments arising from consumption of selenium dietary supplements. Thus, early studies report a decrease in the incidence of certain types of cancer, whereas subsequent studies did not observe any anti-cancer effect, and adverse effects such as increased risks for type 2 diabetes have been reported. A possible contributing factor may be that different chemical forms of selenium were used in different studies. Using larval stage zebrafish (Danio rerio) as a model organism, we report a comparison of the toxicities and tissue selenium distributions of four different chemical forms of selenium. We find that the organic forms of selenium tested (Se-methyl-l-selenocysteine and l-selenomethionine) show considerably more toxicity than inorganic forms (selenite and selenate), and that this appears to be correlated with the level of bioaccumulation. Despite differences in concentrations, the tissue specific pattern of selenium accumulation was similar for the chemical forms tested; selenium was found to be highly concentrated in pigment (melanin) containing tissues especially for the organic selenium treatments, with lower concentrations in eye lens, yolk sac and heart. These results suggest that pigmented tissues might serve as a storage reservoir for selenium.

Toxicology of organic-inorganic hybrid molecules: bio-organometallics and its toxicology.

Bio-organometallics is a research strategy of biology that uses organic-inorganic hybrid molecules. The molecules are expected to exhibit useful bioactivities based on the unique structure formed by interaction between the organic structure and intramolecular metal(s). However, studies on both biology and toxicology of organic-inorganic hybrid molecules have been incompletely performed. There can be two types of toxicological studies of bio-organometallics; one is evaluation of organic-inorganic hybrid molecules and the other is analysis of biological systems from the viewpoint of toxicology using organic-inorganic hybrid molecules. Our recent studies indicate that cytotoxicity of hybrid molecules containing a metal that is nontoxic in inorganic forms can be more toxic than that of hybrid molecules containing a metal that is toxic in inorganic forms when the structure of the ligand is the same. Additionally, it was revealed that organic-inorganic hybrid molecules are useful for analysis of biological systems important for understanding the toxicity of chemical compounds including heavy metals.

Enriched inorganic compounds in diesel exhaust particles induce mitogen-activated protein kinase activation, cytoskeleton instability, and cytotoxicity in human bronchial epithelial cells.

This study assessed the effects of the diesel exhaust particles on ERK and JNK MAPKs activation, cell rheology (viscoelasticity), and cytotoxicity in bronchial epithelial airway cells (BEAS-2B). Crude DEP and DEP after extraction with hexane (DEP/HEX) were utilized. The partial reduction of some DEP/HEX organics increased the biodisponibility of many metallic elements. JNK and ERK were activated simultaneously by crude DEP with no alterations in viscoelasticity of the cells. Mitochondrial activity, however, revealed a decrease through the MTT assay. DEP/HEX treatment increased viscoelasticity and cytotoxicity (membrane damage), and also activated JNK. Our data suggest that the greater bioavailability of metals could be involved in JNK activation and, consequently, in the reduction of fiber coherence and increase in the viscoelasticity and cytotoxicity of BEAS cells. The adverse findings detected after exposure to crude DEP and to DEP/HEX reflect the toxic potential of diesel compounds. Considering the fact that the cells of the respiratory epithelium are the first line of defense between the body and the environment, our data contribute to a better understanding of the pathways leading to respiratory cell injury and provide evidence for the onset of or worsening of respiratory diseases caused by inorganic compounds present in DEP.

DNA damage caused by inorganic particulate matter on Raji and HepG2 cell lines exposed to ultraviolet radiation.

Epidemiological studies have correlated exposure to ultraviolet-irradiated particulate matter with cardiovascular, respiratory, and lung diseases. This study investigated the DNA damage induced by two major inorganic particulate matter compounds found in diesel exhaust, ammonium nitrate and ammonium sulfate, on Burkitt's lymphoma (Raji) and hepatocellular carcinoma (HepG2) cell lines. We found a dose-dependent positive correlation of accumulated DNA damage at concentrations of ammonium nitrate (25 µg/ml, 50 µg/ml, 100 µg/ml, 200 µg/ml, 400 µg/ml) with ultraviolet exposure (250 J/m(2), 400 J/m(2), 600 J/m(2), 850 J/m(2)), as measured by the comet assay in both cell lines. There was a significant difference between the treated ammonium nitrate samples and negative control samples in Raji and HepG2 cells (p<0.001). Apoptosis was shown in Raji and HepG2 cells when exposed to high concentrations of ammonium nitrate (200 µg/ml and 400 µg/ml) for 1h in samples without ultraviolet exposure, as assessed by the comet assay. However, the level of apoptosis greatly diminished after ultraviolet exposure at these concentrations. Over a 24h period, at intervals of 1, 4, 8, 12, 18, and 24h, we also observed that ammonium nitrate decreased viability in Raji and HepG2 cell lines and inhibited cell growth. Ammonium sulfate-induced DNA damage was minimal in both cell lines, but there remained a significant difference (p<0.05) between the ultraviolet radiation treated and negative control samples. These results indicate that the inorganic particulate compound, ammonium nitrate, induced DNA strand breaks at all concentrations, and indications of apoptosis at high concentrations in Raji and HepG2 cells, with ultraviolet radiation preventing apoptosis at high concentrations. We hypothesize that ultraviolet radiation may inhibit an essential cellular mechanism, possibly involving p53, thereby explaining this phenomenon. Further studies are necessary to characterize the roles of apoptosis inhibition induced by DNA damage caused by inorganic particulate matter.

Spore Resistance Properties.

Spores of various Bacillus and Clostridium species are among the most resistant life forms known. Since the spores of some species are causative agents of much food spoilage, food poisoning, and human disease, and the spores of Bacillus anthracis are a major bioweapon, there is much interest in the mechanisms of spore resistance and how these spores can be killed. This article will discuss the factors involved in spore resistance to agents such as wet and dry heat, desiccation, UV and γ-radiation, enzymes that hydrolyze bacterial cell walls, and a variety of toxic chemicals, including genotoxic agents, oxidizing agents, aldehydes, acid, and alkali. These resistance factors include the outer layers of the spore, such as the thick proteinaceous coat that detoxifies reactive chemicals; the relatively impermeable inner spore membrane that restricts access of toxic chemicals to the spore core containing the spore's DNA and most enzymes; the low water content and high level of dipicolinic acid in the spore core that protect core macromolecules from the effects of heat and desiccation; the saturation of spore DNA with a novel group of proteins that protect the DNA against heat, genotoxic chemicals, and radiation; and the repair of radiation damage to DNA when spores germinate and return to life. Despite their extreme resistance, spores can be killed, including by damage to DNA, crucial spore proteins, the spore's inner membrane, and one or more components of the spore germination apparatus.

Toxic inhalational exposures.

Respirable toxicants are a spectrum of irritant and nonirritant gases, vapors, fumes, and airborne particles that can be entrained into the body through the respiratory tract, resulting in exposures that cause pulmonary injury and/or systemic disease. Sources of respirable toxicants include structural fires, industrial accidents, domestic mishaps, and intentional releases of injurious agents on the battleground (warfare) or in civilian settings (acts of terrorism). Acute toxic inhalational exposures may result in respiratory failure, multisystem organ dysfunction, and death. Management of victims includes assessment and protection of the airway, monitoring and treatment of systemic toxicity, and delivery of exposure-specific and nonspecific therapies that improve outcomes. Treatments may include antidotes, hyperbaric oxygen, and other nonspecific life-supporting interventions.

[The impact of ecologic risk factors in mining cities of South Ural].

The analysis of common and primary morbidity according the statistic form 12 is presented. The assessment of health risk in major age groups of population in mining cities of Republic of Bashkortostan is provided. The dynamics of morbidity indicators trends to increasing because in all age groups higher levels of morbidity were detected. The analysis of input of environment pollution into morbidity of population of cities of Utchaly and Sybay revealed that a significant role is played by environment factors. So, the existence of specific geochemical territory and anthropogenic pollution of environment with inorganic compounds of highly toxic metals is a most significant risk fasctor impacting population health. This condition urges to develop various preventive measures.