Bioprospecting of extremophilic perchlorate-reducing bacteria: report of promising Bacillus spp. isolated from sediments of the bay of Cartagena, Colombia.

Three extremophile bacterial strains (BBCOL-009, BBCOL-014 and BBCOL-015), capable of degrading high concentrations of perchlorate at a range of pH (6.5 to 10.0), were isolated from Colombian Caribbean Coast sediments. Morphological features included Gram negative strain bacilli with sizes averaged of 1.75 × 0.95, 2.32 × 0.65 and 3.08 × 0.70 µm, respectively. The reported strains tolerate a wide range of pH (6.5 to 10.0); concentrations of NaCl (3.5 to 7.5% w/v) and KClO4- (250 to 10000 mg/L), reduction of KClO4- from 10 to 25%. LB broth with NaCl (3.5-30% w/v) and KClO4- (250-10000 mg/L) were used in independent trials to evaluate susceptibility to salinity and perchlorate, respectively. Isolates increased their biomass at 7.5 % (w/v) NaCl with optimal development at 3.5 % NaCl. Subsequently, ClO4- reduction was assessed using LB medium with 3.5% NaCl and 10000 mg/L ClO4-. BBCOL-009, BBCOL-014 and BBCOL-015 achieved 10%, 17%, and 25% reduction of ClO4-, respectively. The 16 S rRNA gene sequence grouped them as Bacillus flexus T6186-2, Bacillus marisflavi TF-11 (T), and Bacillus vietnamensis 15 - 1 (T) respectively, with < 97.5% homology. In addition, antimicrobial resistance to ertapenem, vancomycine, amoxicillin clavulanate, penicillin, and erythromycin was present in all the isolates, indicating their high adaptability to stressful environments. The isolated strains from marine sediments in Cartagena Bay, Colombia are suitable candidates to reduce perchlorate contamination in different environments. Although the primary focus of the study of perchlorate-reducing and resistant bacteria is in the ecological and agricultural realms, from an astrobiological perspective, perchlorate-resistant bacteria serve as models for astrobiological investigations.

Genotoxicity biomarkers in car repair workers from Barranquilla, a Colombian Caribbean City.

Exposure to chemicals and particles generated in automotive repair shops is a common and underestimated problem. The objective of this study was to assess the genotoxic status of auto repair workers with (1) a questionnaire to gather sociodemographic information and self-reported exposure to hazardous chemicals and (2) measurement of various biochemical parameters. Blood and oral mucosa samples were collected from 174 male volunteers from Barranquilla, Colombia, aged 18-55 years: 87 were active car repairmen and 87 were individuals with no known exposure to hazardous chemicals. Peripheral blood lymphocytes were collected for the comet and cytokinesis-blocking micronucleus (CBMN) assays, while oral mucosal epithelium extracted to quantify micronucleated cells (MNC). DNA was extracted to assess polymorphisms in the DNA repair (XRCC1) and metabolism-related genes (GSTT1 and GSTM1) using PCR-RFLP. DNA damage and frequency of micronuclei (MN) in lymphocytes and oral mucosa were significantly higher in exposed compared to control group. In both groups genotypes and allelic variants for XRCC1 and GSTT1 met the Hardy-Weinberg equilibrium (HWE). In contrast, GSTM1 deviated from HWE. In the exposed group genotypic variants were not correlated with DNA damage or MN presence in cells. DNA damage and occurrence of MN in mucosa and lymphocytes correlated with age and time of service (occupational exposure ≥ 3 years). In summary, workers in car repair shops exhibited genotoxic effects depending upon exposure duration in the workplace which occurred independent of DNA repair XRCC1 gene and metabolism genes GSTT1 and GSTM1. Date demonstrate that health authorities improve air quality in auto repair facilities to avoid occupational DNA damage.

Perchlorate Contamination: Sources, Effects, and Technologies for Remediation.

Perchlorate is a persistent pollutant, generated via natural and anthropogenic processes, that possesses a high potential for endocrine disruption in humans and biota. It inhibits iodine fixation, a major reason for eliminating this pollutant from ecosystems. Remediation of perchlorate can be achieved with various physicochemical treatments, especially at low concentrations. However, microbiological approaches using microorganisms, such as those from the genera Dechloromonas, Serratia, Propionivibrio, Wolinella, and Azospirillum, are promising when perchlorate pollution is extensive. Perchlorate-reducing bacteria, isolated from harsh environments, for example saline soils, mine sediments, thermal waters, wastewater treatment plants, underground gas storage facilities, and remote areas, including the Antarctica, can provide removal yields from 20 to 100%. Perchlorate reduction, carried out by a series of enzymes, such as perchlorate reductase and superoxide chlorite, depends on pH, temperature, salt concentration, metabolic inhibitors, nutritional conditions, time of contact, and cellular concentration. Microbial degradation is cost-effective, simple to implement, and environmentally friendly, rendering it a viable method for alleviating perchlorate pollution in the environment.

Molecular Human Targets of Bioactive Alkaloid-Type Compounds from Tabernaemontana cymose Jacq.

Alkaloids are a group of secondary metabolites that have been widely studied for the discovery of new drugs due to their properties on the central nervous system and their anti-inflammatory, antioxidant and anti-cancer activities. Molecular docking was performed for 10 indole alkaloids identified in the ethanol extract of Tabernaemontana cymosa Jacq. with 951 human targets involved in different diseases. The results were analyzed through the KEGG and STRING databases, finding the most relevant physiological associations for alkaloids. The molecule 5-oxocoronaridine proved to be the most active molecule against human proteins (binding energy affinity average = -9.2 kcal/mol) and the analysis of the interactions between the affected proteins pointed to the PI3K/ Akt/mTOR signaling pathway as the main target. The above indicates that indole alkaloids from T. cymosa constitute a promising source for the search and development of new treatments against different types of cancer.

Emerging contaminants and priority substances in marine sediments from Cartagena Bay and the Grand Marsh of Santa Marta (Ramsar site), Colombia.

Emerging pollutants and priority substances are of growing concern due to their toxicity potential to aquatic organisms and human health. However, few reports on this issue in marine ecosystems in general and, more specifically, on the Colombian Caribbean coast are available. The aim of this study was to detect these compounds in sediments from Cartagena Bay (CB) and in the Grand Marsh of Santa Marta, GMSM (Ramsar site), in order to determine how they related to in vitro cytotoxicity assays on HepG2 cells of sediment extracts. A total of thirty compounds were detected using GC-MS/MS in fifteen stations during both the rainy and the dry seasons. Sediments from CB had a wide range of different toxicants, with polycyclic aromatic hydrocarbons (PAHs) being the most prevalent (12 PAHs, 5.5-881.6 ng/g). Total PCBs ranged from < LOD to 18.6 ng/g, with PCB 138 being the most common detected congener. Residues of p,p'-DDE, Chlorpyrifos and two organophosphate flame retardants, TEHP and ToTP, were found in most sampling locations. The UV filters 4MBC and homosalate were recurrently found in sediments, and the fragrance galaxolide appeared in all cases, with the greatest concentrations found on a touristic beach. In GMSM, with the exception of deltamethrin, all chemicals evaluated had lower average values than in CB. According to sediment quality guidelines, some sites in CB presented values of PAHs higher than the threshold effects level, while in the marsh, none of the stations exceeded it. HepG2 cells exposed to 1% sediment extracts presented reduced cell viability up to 26%. Cytotoxicity displayed a negative correlation with chlorpyrifos concentration. In short, these data suggest the bay and the marsh have specific contamination fingerprints related to anthropogenic interventions. This research highlights the need to further investigate the ecotoxicological implications of detected chemical stressors in these ecosystems.

Intergenerational effects of coal dust on Tribolium castaneum, Herbst.

Coal dust is a primary air contaminant from coal mining operations that produces harmful health effects. However, it is unclear to what extent its detrimental properties would impact future generations, and whether alterations in the progenies might be concentration-dependent. The aim of this study was to determine the intergenerational effects of chronic exposure to coal dust on the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), at three life stages. Groups of fifty adult insects were exposed during 30 days at different concentrations of coal dust mixed with ground oats as food substrate (0, 0.25, 0.5, 1.0 and 2.0% weight/weight), both with a particle size <38 µm. The LC50 for F0 insects was 1.07%, whereas for larvae and adults from F1, values were 0.53 and 0.89%, respectively. Pathological findings assessed at F1 revealed a coal dust concentration-dependent frequency of several morphological abnormalities, including larvae without antenna or extremities, lack of T1, T2, T3 legs, loss of urogomphi, and the presence of abnormal protuberances. It was found that a considerable number of F1 larvae derived from parental beetles did not achieve a complete conversion into the next growth stage. Pupae with undeveloped eyes and adults with malformed elytra, as well as necrosis, were recurrently observed at high concentrations. Finally, adults exposed to 1% coal dust overexpressed genes related to oxidative stress (nuclear factor erythroid 2-related factor 2, Nrf2) and synaptic transmission (GABA-gated ion channel, Grd). In short, coal dust particles induced intergenerational effects on T. castaneum, highlighting the need to further study the impact of this airborne pollutant on wildlife and human populations.

Intergenerational toxicity of nonylphenol ethoxylate (NP-9) in Caenorhabditis elegans.

The ethoxylated isomers of nonylphenol (NPEs, NP-9) are one of the main active ingredients present in nonionic surfactants employed as herbicides, cosmetics, paints, plastics, disinfectants and detergents. These chemicals and their metabolites are commonly found in environmental matrices. The aim of this work was to evaluate the intergenerational toxicity of NP-9 in Caenorhabditis elegans. The lethality, length, width, locomotion and lifespan were investigated in the larval stage L4 of the wild strain N2. Transgenic green fluorescent protein (GFP) strains were employed to estimate changes in relative gene expression. RT-qPCR was utilized to measure mRNA expression for neurotoxicity-related genes (unc-30, unc-25, dop-3, dat-1, mgl-1, and eat-4). Data were obtained from parent worms (P0) and the first generation (F1). Lethality of the nematode was concentration-dependent, with 48 h-LC50 values of 3215 and 1983 µM in P0 and F1, respectively. Non-lethal concentrations of NP-9 reduced locomotion. Lifespan was also decreased by the xenobiotic, but the negative effect was greater in P0 than in F1. Non-monotonic concentration-response curves were observed for body length and width in both generations. The gene expression profile in P0 was different from that registered in F1, although the expression of sod-4, hsp-70, gpx-6 and mtl-2 increased with the surfactant concentration in both generations. None of the tested genes followed a classical concentration-neurotoxicity relationship. In P0, dopamine presented an inverted-U curve, while GABA and glutamate displayed a bimodal type. However, in F1, inverted U-shaped curves were revealed for these genes. In summary, NP-9 induced intergenerational responses in C. elegans through mechanisms involving ROS, and alterations of the GABA, glutamate, and dopamine pathways.

Toxicity of nonylphenol and nonylphenol ethoxylate on Caenorhabditis elegans.

Among the most used chemicals in the world are nonionic surfactants. One of these environmental pollutants is nonylphenol ethoxylate (NP-9), also known as Tergitol, and its degradation product, nonylphenol (NP). The objective of this work was to determine the toxicity of NP and NP-9 in Caenorhabditis elegans. Wild-type L4 larvae were exposed to different concentrations of the surfactants to measure functional endpoints. Mutant strains were employed to promote the activation of toxicity signaling pathways related to mtl-2, gst-1, gpx-4, gpx-6, sod-4, hsp-70 and hsp-4. Additionally, stress response was also assessed using a daf-16::GFP transgenic strain. The lethality was concentration dependent, with 24-h LC50 of 122 µM and 3215 µM for NP and NP-9, respectively. Both compounds inhibited nematode growth, although NP was more potent; and at non-lethal concentrations, nematode locomotion was reduced. The increase in the expression of tested genes was significant at 10 µM for NP-9 and 0.001 µM for NP, implying a likely role for the activation of oxidative and cellular stress, as well as metabolism pathways. With the exception of glutathione peroxidase, which has a bimodal concentration-response curve for NP, typical of endocrine disruption, the other curves for this xenobiotic in the strains evaluated were almost flat for most concentrations, until reaching 50-100 µM, where the effect peaked. NP and NP-9 induced the activation and nuclear translocation of DAF-16, suggesting that transcription of stress-response genes may be mediated by the insulin/IGF-1 signaling pathway. In contrast, NP-9 induced a concentration-dependent response for the sod-4 and hsp-4 mutants, with greater fluorescence induction than NP at similar levels. In short, NP and NP-9 affect the physiology of C. elegans and modulate gene expression related to ROS production, cellular stress and metabolism of xenobiotics.

Biomonitoring of Mercury, Cadmium and Selenium in Fish and the Population of Puerto Nariño, at the Southern Corner of the Colombian Amazon.

Heavy metals threaten communities near biodiversity hotspots, as their protein sources come from the environment. This study assessed Hg, Cd, and Se concentrations in fish, as well as the magnitude of exposure and hematological conditions of adult citizens from Puerto Nariño (Colombian Amazon). Among fish samples, greater Hg concentrations were found in higher trophic level species, including Rhaphiodon vulpinus (880 ± 130 ng/g) and Pseudoplatystoma tigrinum (920 ± 87 ng/g). These species presented the highest hazard quotients and lowest Se:Hg molar ratios among those studied, showing their consumption represents a health risk to consumers. Moreover, some samples of Mylossoma duriventre and Prochilodus magdalenae had Cd levels greater than the regulated limit (100 ng/g). The average total Hg (T-Hg) concentrations in human hair and blood were 5.31 µg/g and 13.7 µg/L, respectively. All hair samples exceeded the 1.0 µg/g threshold set by the USEPA, whereas 93% of the volunteers had T-Hg blood levels greater than 5 µg/L, suggesting elevated exposure. The mean Cd level was 3.1 µg/L, with 21% of samples surpassing 5 µg/L, value at which mitigating actions should be taken. Eighty-four percent of participants presented Se deficiencies (<100 µg/L). There was a significant association between fish consumption and T-Hg in hair (ρ = 0.323; p = 0.032) and blood (ρ = 0.381; p = 0.011). In this last matrix, Se correlated with Cd content, whereas lymphocytes were inversely linked to Hg concentrations. The results of this study show that there is  extensive exposure to Hg in fish, the consumption of which may promote detrimental impacts on hematology parameters within the community.

Proteasome-associated cysteine deubiquitinases are molecular targets of environmental optical brightener compounds.

The levels of organic pollutants, such as optical brightener (OB) compounds, in the global environment have been increasing in recent years. The toxicological effects and signal transduction systems associated with OB toxicity have not been thoroughly studied. The ubiquitin-proteasome system (UPS) plays a crucial role in regulating multiple essential cellular processes, and proteasome-associated cysteine deubiquitinases (DUBs), ubiquitin C-terminal hydrolase L5 (UCHL5) and USP14, are two major regulators for (de)ubiquitination and stability of many important target proteins. Therefore, potential inhibition of UCHL5 and USP14 activities by some environmental chemicals might cause in vivo toxicity. In the current study we hypothesize that electrophilic OB compounds, such as 4,4'-diamino-2,2'-stilbenedisulfonic acid(DAST), fluorescent brightener 28 (FB-28) and FB-71, can interact with the catalytic triads (CYS, HIS, and ASP) of UCHL5 and USP14 and inhibit their enzymatic activities, leading to cell growth suppression. This hypothesis is supported by our findings presented in this study. Results from in silico computational docking and ubiquitin vinyl sulfone assay confirmed the UCHL5/USP14-inhibitory activities of these OB compounds that have potencies in an order of: FB-71 > FB-28 > DAST. Furthermore, inhibition of these two proteasomal DUBs by OBs resulted in cell growth inhibition and apoptosis induction in two human breast cancer cell models. In addition, we found that OB-mediated DUB inhibition triggers a feedback reaction in which expression of UCHL5 and USP14 proteins is increased to compromise the suppressed activities. Our study suggests that these commonly used OB compounds may target and inhibit proteasomal cysteine DUBs, which should contribute to their toxicological effects in vivo.

Toxicity and expression of oxidative stress genes in Tribolium castaneum induced by toluene, xylene, and thinner.

Thinner is a solvent that contains toluene, xylene, and other substances. The aim of this study was to examine the toxicity of thinner, xylene, and toluene on adult red flour beetles, Tribolium castaneum (Herbst), as well as the effects of thinner and toluene on gene expression. The major compounds identified in the thinner with peak area >2% were p-xylene (6%), toluene (4%), 2,4-dimethylheptane (3%), methylcyclohexane (2.75%), 2-methylheptane (2%), cyclohexanone (2.6%), and nonane (2.1%). Insects were exposed to solvents at 0-240 µl/L air for 4-48 hr. Animals that survived after a 4 hr exposure to 80 µl/L air were employed to determine mRNA expression using real-time PCR. Xylene was highly toxic, and the majority of the beetles did not survive 4 hr exposure at 40 µl/L air. The lethal concentration 50 (LC50) values for toluene, xylene and thinner at 48 hr exposure were 97.7, <40 and 99.8 µl/L air, respectively. Thinner and toluene induced lethargy and sluggish movement in treated insects, while with xylene these effects were not observed. Glutathione-S-transferase (Gst) gene expression increased after thinner and toluene exposure. Superoxide dismutase (SOD) gene expression rose after toluene exposure. These results suggest that T. castaneum provides an alternative model for the study of toxicological effects of volatile aromatic compounds.

Multicompartment Mercury Contamination in Major Gold Mining Districts at the Department of Bolivar, Colombia.

Artisanal and small-scale gold mining is the main source of human exposure to mercury (Hg) in many countries. This study was designed to evaluate total Hg (T-Hg) concentrations in human hair, fish, soil, and air from two major gold-mining districts (GMDs) at the department of Bolivar, Colombia. Total Hg was analyzed using a direct Hg analyzer. The mean T-Hg concentration in hair samples was 3.07 ± 0.14 µg/g (range 0.15-25.1 µg/g; median 2.02 µg/g). The highest Hg level was observed in Mojana GMD, specifically at Achi-La Raya (9.2 ± 0.6 µg/g) and the lowest in Morales, at the Middle Magdalena GMD (1.50 ± 0.16 µg/g). Hair T-Hg values exceeded the U.S. Environmental Protection Agency reference level of 1.0 µg/g. Correlation between T-Hg in hair and stature was negative for the Mojana, but the opposite for Middle Magdalena, although for both GMDs hair T-Hg correlated positively with fish intake. The highest average T-Hg fish concentrations were observed in Caquetaia kraussii (0.37 ± 0.10 µg/g), Sorubim cuspicaudus (0.32 ± 0.16 µg/g), Plagioscion surinamensis (0.22 ± 0.02 µg/g), Trachelyopterus insignis (0.20 ± 0.02 µg/g), and Pseudoplatystoma magdaleniatum (0.20 ± 0.02 µg/g). Human health risk assessment of Hg based on fish consumption suggested that, with the exception of P. magdalenae, all economically important fish species are potentially harmful for the communities. Soil Hg levels in amalgam burning facilities were extremely high, and Hg in the air around mines and gold-processing shops exceeded international guidelines. In short, Hg pollution in GMDs of Bolivar is extensive, and this situation requires special attention to reduce environmental and human health problems.

Toxicity of atrazine- and glyphosate-based formulations on Caenorhabditis elegans.

Atrazine and Glyphosate are herbicides massively used in agriculture for crop protection. Upon application, they are available to the biota in different ecosystems. The aim of this research was to evaluate the toxicity of Glyphosate and Atrazine based formulations (GBF and ABF, respectively). Caenorhabditis elegans was exposed to different concentrations of each single formulation, and to the mixture. Lethality, locomotion, growth, and fertility were measured as endpoints. Effects on gene expression were monitored utilizing green fluorescence protein transgenic strains. ABF caused lethality of 12%, 15%, and 18% for 6, 60, and 600 µM, respectively, displaying a dose dependence trend. GBF produced lethality of 20%, 50%, and 100% at 0.01, 10, and 100 µM, respectively. Locomotion inhibition ranged from 21% to 89% at the lowest and maximum tested concentrations for Atrazine; whereas for Glyphosate, exposure to 10 µM inhibited 87%. Brood size was decreased by 67% and 93% after treatment to 0.06 and 6 µM Atrazine, respectively; and by 23% and 93% after exposure to 0.01 and 10 µM Glyphosate, respectively. There were no significant differences in growth. Changes in gene expression occurred in all genes, highlighting the expression of sod-1, sod-4, and gpx-4 that increased more than two-fold after exposure to 600 µM ABF and 10 µM GBF. The effects observed for the mixture of these formulations were additive for lethality, locomotion and fertility. In short, GBF, ABF, and their mixture induced several toxic responses related to oxidative stress on C. elegans.

Bioecological Aspects and Heavy Metal Contamination of the Mollusk Donax denticulatus in the Colombian Caribbean Coastline.

Donax denticulatus is a key mollusk for the ecology of sandy beaches, serving as a controller of organic matter, microorganisms, and as bioindicator of heavy metals pollution. The goal of this study was to characterize some ecological aspects of D. denticulatus and its relationship with the content of heavy metals in their tissue, in three beaches of the Caribbean coast of Colombia. The results showed the study populations were different in terms of morphological characteristics and density (Berrugas-Sucre < Cartagena-Bolívar < Riohacha-Guajira), but not in sexual proportion; although density was clearly related to beach occupancy by tourists. Analysis of metals revealed tissue concentrations varied depending on the location (Greater means: Hg = 0.018 ± 0.004 in Riohacha; Pb = 0.110 ± 0.060 in Berrugas and Cd = 0.040 ± 0.010 µg/g in Cartagena). No relationships were found between morphometric variables and heavy metals content. Principal components analysis highlighted Riohacha for presenting differences respecting to Bocagrande and Berrugas in terms of physicochemical water parameters such as pH, temperature, salinity, dissolved oxygen and conductivity. Results suggest tourism rather than environmental pollution could be a sensitive factor for biota survival in Caribbean beaches.

Natural Products as Chemopreventive Agents by Potential Inhibition of the Kinase Domain in ErbB Receptors.

Small molecules found in natural products provide therapeutic benefits due to their pharmacological or biological activity, which may increase or decrease the expression of human epidermal growth factor receptor (HER), a promising target in the modification of signaling cascades involved in excessive cellular growth. In this study, in silico molecular protein-ligand docking protocols were performed with AutoDock Vina in order to evaluate the interaction of 800 natural compounds (NPs) from the NatProd Collection (http://www.msdiscovery.com/natprod.html), with four human HER family members: HER1 (PDB: 2ITW), HER2 (PDB: 3PP0), HER3 (PDB: 3LMG) and HER4 (PDB: 2R4B). The best binding affinity values (kcal/mol) for docking pairs were obtained for HER1-podototarin (-10.7), HER2-hecogenin acetate (-11.2), HER3-hesperidin (-11.5) and HER4-theaflavin (-10.7). The reliability of the theoretical calculations was evaluated employing published data on HER inhibition correlated with in silico binding calculations. IC50 values followed a significant linear relationship with the theoretical binding Affinity data for HER1 (R = 0.656, p < 0.0001) and HER2 (R = 0.543, p < 0.0001), but not for HER4 (R = 0.364, p > 0.05). In short, this methodology allowed the identification of several NPs as HER inhibitors, being useful in the discovery and design of more potent and selective anticancer drugs.

Neurotoxic Effects of Linalool and ß-Pinene on Tribolium castaneum Herbst.

Effective, ethical pest control requires the use of chemicals that are highly specific, safe, and ecofriendly. Linalool and ß-pinene occur naturally as major constituents of the essential oils of many plant species distributed throughout the world, and thus meet these requirements. These monoterpenes were tested as repellents against Tribolium castaneum, using the area preference method, after four hours of exposure and the effect transcriptional of genes associated with neurotransmission. Changes in gene expression of acetylcholinesterase (Ace1), GABA-gated anion channel splice variant 3a6a (Rdl), GABA-gated ion channel (Grd), glutamate-gated chloride channel (Glucl), and histamine-gated chloride channel 2 (Hiscl2) were assessed and the interaction with proteins important for the insect using in silico methods was also studied. For linalool and ß-pinene, the repellent concentration 50 (RC50) values were 0.11 µL/cm² and 0.03 µL/cm², respectively. Both compounds induced overexpression of Hiscl2 gen in adult insects, and ß-pinene also promoted the overexpression of Grd and the Ace1 gene. However, ß-pinene and linalool had little potential to dock on computer-generated models for GABA-gated ion channel LCCH3, nicotinic acetylcholine receptor subunits alpha1 and alpha2, and putative octopamine/tyramine receptor proteins from T. castaneum as their respective binding affinities were marginal, and therefore the repellent action probably involved mechanisms other than direct interaction with these targets. Results indicated that ß-pinene was more potent than linalool in inducing insect repellency, and also had a greater capacity to generate changes in the expression of genes involved in neuronal transmission.

Mice housed on coal dust-contaminated sand: A model to evaluate the impacts of coal mining on health.

Coal dust is the most important air pollutant in coal mining in regards to producing deleterious health effects. It permeates the surrounding environment threatening public health. The aim of this study was to evaluate the toxic effects associated with exposure to sand contaminated with coal dust particles below 38 µm in diameter, obtained from a mineral sample collected in the largest coal mine in South America, La Loma, Cesar, Colombia. Sterilized sand was spiked with coal dust to obtain concentrations ranging from zero to 4% coal dust. To model natural exposure, mice were housed for eight weeks in boxes containing this mixture as bedding after which, they were euthanized and blood and tissue samples were collected. Real time PCR analysis revealed an increase in Cyp1A1 mRNA for living on sand with coal dust concentrations greater than 2% compared to mice living on sand without coal dust. Unexpectedly, for mice on coal dust-polluted sand, Sod1, Scd1 and Nqo1 hepatic mRNA were downregulated. The Comet assay in peripheral blood cells and the micronucleus test in blood smears, showed a significant potential genotoxic effect only at the highest coal dust concentration. Histopathological analysis revealed vascular congestion and peribronchial inflammation in the lungs. A dose-response relationship for the presence of hepatic steatosis, vacuolization and nuclei enlargements was observed in the exposed animals. The data suggest living on a soil polluted with coal dust induces molecular, cellular and histopathological changes in mice. Accordingly, the proposed model can be used to identify deleterious effects of exposure to coal dust deposited in soils that may pose health risks for surrounding wildlife populations.

Urban Endocrine Disruptors Targeting Breast Cancer Proteins.

Humans are exposed to a huge amount of environmental pollutants called endocrine disrupting chemicals (EDCs). These molecules interfere with the homeostasis of the body, usually through mimicking natural hormones leading to activation or blocking of their receptors. Many of these compounds have been associated with a broad range of diseases including the development or increased susceptibility to breast cancer, the most prevalent cancer in women worldwide, according to the World Health Organization. Thus, this article presents a virtual high-throughput screening (vHTS) to evaluate the affinity of proteins related to breast cancer, such as ESR1, ERBB2, PGR, BCRA1, and SHBG, among others, with EDCs from urban sources. A blind docking strategy was employed to screen each protein-ligand pair in triplicate in AutoDock Vina 2.0, using the computed binding affinities as ranking criteria. The three-dimensional structures were previously obtained from EDCs DataBank and Protein Data Bank, prepared and optimized by SYBYL X-2.0. Some of the chemicals that exhibited the best affinity scores for breast cancer proteins in each category were 1,3,7,8-tetrachlorodibenzo-p-dioxin, bisphenol A derivatives, perfluorooctanesulfonic acid, and benzo(a)pyrene, for catalase, several proteins, sex hormone-binding globulin, and cytochrome P450 1A2, respectively. An experimental validation of this approach was performed with a complex that gave a moderate binding affinity in silico, the sex hormone binding globulin (SHBG), and bisphenol A (BPA) complex. The protein was obtained using DNA recombinant technology and the physical interaction with BPA assessed through spectroscopic techniques. BPA binds on the recombinant SHBG, and this results in an increase of its α helix content. In short, this work shows the potential of several EDCs to bind breast cancer associated proteins as a tool to prioritize compounds to perform in vitro analysis to benefit the regulation or exposure prevention by the general population.

Caenorhabditis elegans, a Biological Model for Research in Toxicology.

Caenorhabditis elegans is a nematode of microscopic size which, due to its biological characteristics, has been used since the 1970s as a model for research in molecular biology, medicine, pharmacology, and toxicology. It was the first animal whose genome was completely sequenced and has played a key role in the understanding of apoptosis and RNA interference. The transparency of its body, short lifespan, ability to self-fertilize and ease of culture are advantages that make it ideal as a model in toxicology. Due to the fact that some of its biochemical pathways are similar to those of humans, it has been employed in research in several fields. C. elegans' use as a biological model in environmental toxicological assessments allows the determination of multiple endpoints. Some of these utilize the effects on the biological functions of the nematode and others use molecular markers. Endpoints such as lethality, growth, reproduction, and locomotion are the most studied, and usually employ the wild type Bristol N2 strain. Other endpoints use reporter genes, such as green fluorescence protein, driven by regulatory sequences from other genes related to different mechanisms of toxicity, such as heat shock, oxidative stress, CYP system, and metallothioneins among others, allowing the study of gene expression in a manner both rapid and easy. These transgenic strains of C. elegans represent a powerful tool to assess toxicity pathways for mixtures and environmental samples, and their numbers are growing in diversity and selectivity. However, other molecular biology techniques, including DNA microarrays and MicroRNAs have been explored to assess the effects of different toxicants and samples. C. elegans has allowed the assessment of neurotoxic effects for heavy metals and pesticides, among those more frequently studied, as the nematode has a very well defined nervous system. More recently, nanoparticles are emergent pollutants whose toxicity can be explored using this nematode. Overall, almost every type of known toxicant has been tested with this animal model. In the near future, the available knowledge on the life cycle of C. elegans should allow more studies on reproduction and transgenerational toxicity for newly developed chemicals and materials, facilitating their introduction in the market. The great diversity of endpoints and possibilities of this animal makes it an easy first-choice for rapid toxicity screening or to detail signaling pathways involved in mechanisms of toxicity.

Toxicogenomics to Evaluate Endocrine Disrupting Effects of Environmental Chemicals Using the Zebrafish Model.

The extent of our knowledge on the number of chemical compounds related to anthropogenic activities that can cause damage to the environment and to organisms is increasing. Endocrine disrupting chemicals (EDCs) are one group of potentially hazardous substances that include natural and synthetic chemicals and have the ability to mimic endogenous hormones, interfering with their biosynthesis, metabolism, and normal functions. Adverse effects associated with EDC exposure have been documented in aquatic biota and there is widespread interest in the characterization and understanding of their modes of action. Fish are considered one of the primary risk organisms for EDCs. Zebrafish (Danio rerio) are increasingly used as an animal model to study the effects of endocrine disruptors, due to their advantages compared to other model organisms. One approach to assess the toxicity of a compound is to identify those patterns of gene expression found in a tissue or organ exposed to particular classes of chemicals, through new technologies in genomics (toxicogenomics), such as microarrays or whole-genome sequencing. Application of these technologies permit the quantitative analysis of thousands of gene expression changes simultaneously in a single experiment and offer the opportunity to use transcript profiling as a tool to predict toxic outcomes of exposure to particular compounds. The application of toxicogenomic tools for identification of chemicals with endocrine disrupting capacity using the zebrafish model system is reviewed.