Oxidative stress and alterations in the expression of genes related to inflammation, DNA damage, and metal exposure in lung cells exposed to a hydroethanolic coal dust extract.

BACKGROUND: Open cast mining is well known as a concerning source of environmental and public health problems. This work aimed to obtain a hydroethanolic coal dust extract (≤ 38 µm) and to characterize its composition with particular regard to content of organic compounds by GC/MS, as well as describe its toxicity in vitro on Calu-1 after exposure to several concentrations (0-500 µg/mL). MATERIALS AND RESULTS: Cytotoxicity was measured with MTT assay and DCFH-DA probe was employed to estimate the amount of reactive oxygen species (ROS) in Calu-1 cells. RT-PCR was employed to quantify relative expression of genes associated with inflammation, oxidative stress, as well as metals, and lipid metabolism. Seventeen organic compounds were identified in the extract, highlighting undecane, dodecane, pentadecane and benzo[a]anthracene, 6,12-dimethyl-1,2,3,4-tetrahydro-. Cytotoxicity test showed a decrease trend in the cell viability after 24 h hours from the concentration of 62.5 µg/mL. Further, the extract raised intracellular ROS when compared with control. Expression levels of CYP1A1, IL-8, IL-6, MT1X, and NQO1 were up-regulated when cells were exposed to 125 µg/mL of coal dust, whereas PPAR-α was down-regulated, likely involving aryl hydrocarbon receptor regulation. CONCLUSIONS: In short, this study shows that despite hydroethanolic coal dust extract is not cytotoxic to Calu-1 cells, it produces an elevation of intracellular ROS and alters the expression in marker genes of oxidative stress, inflammation, metal transport, xenobiotic and lipid metabolism. These findings suggest that chemicals present in coal dust are biologically active and may interfere key biochemical process in the living organisms.

Toxicological effects of bituminous coal dust on the earthworm Eisenia fetida (Oligochaeta: Lumbricidae).

The exploitation of coal is an important resource to generate energy worldwide. However, during the processes of coal extraction, transport, and cargo, dust particles are released into the environment. The aim of this study was to determine the toxicological effects of bituminous coal dust (<38 µm), obtained from a sample collected in a coal mine in Colombia, on the annelid Eisenia fetida. The earthworm culture was standardized under laboratory conditions to evaluate mortality, as well as morphological, physiological and histological changes using concentrations varying from 1 to 4% w/w coal dust in artificial soil, after 7, 14, and 28 days of exposure. In addition, an avoidance assay was carried out after 48-h treatment. Histopathological analysis was performed at the end of the experiment. After the sub-chronic exposure, an increase in mortality was observed at the highest coal dust concentration compared to the untreated group. Alterations in morphology and physiology of the exposed annelids were mostly evidenced at the greatest tested concentrations (3-4%) and exposure times (≥14 days). Changes included loss of weight and color, abundant mucus production, constriction, peeling of the epidermis, clitellum involution, violent movements and lethargy. Avoidance of coal dust-polluted soil followed a concentration-response relationship. Histopathological findings revealed changes on the cuticle, as well as in the circular and longitudinal muscle layers in animals living in soils containing 3 and 4% coal particles. In short, E. fetida exposed to coal dust experienced several pathological changes, suggesting that this pollutant may induce population problems in macroinvertebrates present in coal mining areas.

Prediction of the potential biological activity of novel spiropyrazolo[3,4-b]pyridines and spiropyrazolo[3,4-b]pyridine-5,5'-pyrimidines by a ligand-protein inverse-docking approach.

The interaction of a series of spiropyrazolo[3,4-b]pyridines and spiropyrazolo[3,4-b]pyridine-5,5'-pyrimidines with 975 molecular targets involved in different diseases and biochemical alterations in humans was assessed. In-silico and in-vivo methods were used to predict the potential biological activity of these compounds. The exposure of several individuals of C. elegans to these compounds shows that their lethality would be less than 10% and that they do not induce any alteration in their locomotion. The compounds identified as PRV-8 and 13-G were the most bioactive, and also showed other advantages such as; better structural properties, adequate pharmacokinetic and pharmacodynamic properties, and good flexibility and unsaturation, which placed them as the compounds of greatest interest to be tested in-vitro and in-vivo. The series of compounds described here exhibited significant interactions with the estrogen signaling pathway.

In silico evaluation of pesticides as potential modulators of human DNA methyltransferases.

DNA methylations are carried out by DNA methyltransferases (DNMTs) that are key enzymes during gene expression. Many chemicals, including pesticides, have shown modulation of epigenetic functions by inhibiting DNMTs. In this work, human DNMTs were evaluated as a potential target for pesticides through virtual screening of 1038 pesticides on DNMT1 (3SWR) and DNMT3A (2QRV). Molecular docking calculations for DNMTs-pesticide complexes were performed using AutoDock Vina. Binding-affinity values and contact patterns were employed as selection criteria of pesticides as virtual hits for DNMTs. The best three DNMT-pesticides complexes selected according to their high absolute affinity values (kcal/mol), for both DNMT1 and DNMT3A, were flocoumafen (-12.5; -9.9), brodifacoum (-12.4; -8.4) and difenacoum (-12.1; -8.7). These chemicals belong to second-generation rodenticides. The most frequent predicted interacting residues for DNMT1-pesticide complexes were Trp1170A, Phe1145A, Asn1578A, Arg1574A and Pro1225A; whereas for DNMT3A those were Arg271B, Lys740A, and Glu303B. These results suggest that rodenticides used for pest control are potential DNMT ligands and therefore, may modulate DNA methylations. This finding has important environmental and clinical implications, as epigenetic pathways are critical in many biochemical processes leading to diseases.

Imposex in Stramonita haemastoma from coastal sites of Cartagena, Colombia / Imposex em Stramonita haemastoma em locais costeiros de Cartagena, Colômbia

Abstract Imposex is the development of male sexual characteristics caused by the toxic effects of some chemicals that acts as an endocrinal disruptor. Antifouling paints contain these chemicals. Cartagena lacks studies to indicate the extent of imposex in its coastal waters. The aim of this study was to determine the prevalence of imposex in the gastropod Stramonita haemastoma in Cartagena, Colombia. Specimens were collected during 2013 from locations of high and low influence of port activity. Morphometric measurements and the frequency of the occurrence of imposex were registered. The comparison among morphometric variables showed statistically significant differences between the two sites studied. Furthermore, the females of the S. haemastoma species presented an imposex frequency of 93.1% in Birds' Island, Cartagena Bay, compared to 31.8% in La Bocana. The relative penis size index or RPLI (10.145 and 3.231) and vas deferens sequence index or VDSI (2.83 and 1.16), showed possible contamination by organotin compounds in both places.

Resumo Imposex é o desenvolvimento de características sexuais masculinas causadas por poluentes tóxicos de alguns produtos químicos que atuam como desreguladores endócrinos. Tintas anti-incrustantes são as que contêm estes produtos químicos. Cartagena carece de estudos para indicar a extensão do imposex nas suas águas costeiras. O objetivo deste estudo foi determinar a prevalência de imposex no gastrópode Stramonita haemastoma em Cartagena, Colômbia. Os espécimes foram coletados durante 2013 de locais de alta e baixa influência da atividade portuária. Foram registradas as medidas morfométricas e a frequência da ocorrência do imposex. A comparação entre as variáveis morfométricas mostrou diferenças estatisticamente significantes entre os dois locais estudados. Além disso, as fêmeas da espécie S. haemastomaapresentaram uma frequência de imposex de 93,1% na Ilha das Aves, Baía das Cartagena, em comparação com 31,8% em La Bocana. O índice do comprimento relativo do pênis ou RPLI (10,145 e 3,231) e o índice da sequência do vaso deferente ou VDSI (2,83 e 1,16), mostraram possível contaminação por compostos organoestânicos em ambos os locais.

Imposex in Stramonita haemastoma from coastal sites of Cartagena, Colombia.

Imposex is the development of male sexual characteristics caused by the toxic effects of some chemicals that acts as an endocrinal disruptor. Antifouling paints contain these chemicals. Cartagena lacks studies to indicate the extent of imposex in its coastal waters. The aim of this study was to determine the prevalence of imposex in the gastropod Stramonita haemastoma in Cartagena, Colombia. Specimens were collected during 2013 from locations of high and low influence of port activity. Morphometric measurements and the frequency of the occurrence of imposex were registered. The comparison among morphometric variables showed statistically significant differences between the two sites studied. Furthermore, the females of the S. haemastoma species presented an imposex frequency of 93.1% in Birds' Island, Cartagena Bay, compared to 31.8% in La Bocana. The relative penis size index or RPLI (10.145 and 3.231) and vas deferens sequence index or VDSI (2.83 and 1.16), showed possible contamination by organotin compounds in both places.

Imposex in Stramonita haemastoma from coastal sites of Cartagena, Colombia

Abstract Imposex is the development of male sexual characteristics caused by the toxic effects of some chemicals that acts as an endocrinal disruptor. Antifouling paints contain these chemicals. Cartagena lacks studies to indicate the extent of imposex in its coastal waters. The aim of this study was to determine the prevalence of imposex in the gastropod Stramonita haemastoma in Cartagena, Colombia. Specimens were collected during 2013 from locations of high and low influence of port activity. Morphometric measurements and the frequency of the occurrence of imposex were registered. The comparison among morphometric variables showed statistically significant differences between the two sites studied. Furthermore, the females of the S. haemastoma species presented an imposex frequency of 93.1% in Birds Island, Cartagena Bay, compared to 31.8% in La Bocana. The relative penis size index or RPLI (10.145 and 3.231) and vas deferens sequence index or VDSI (2.83 and 1.16), showed possible contamination by organotin compounds in both places.

Resumo Imposex é o desenvolvimento de características sexuais masculinas causadas por poluentes tóxicos de alguns produtos químicos que atuam como desreguladores endócrinos. Tintas anti-incrustantes são as que contêm estes produtos químicos. Cartagena carece de estudos para indicar a extensão do imposex nas suas águas costeiras. O objetivo deste estudo foi determinar a prevalência de imposex no gastrópode Stramonita haemastoma em Cartagena, Colômbia. Os espécimes foram coletados durante 2013 de locais de alta e baixa influência da atividade portuária. Foram registradas as medidas morfométricas e a frequência da ocorrência do imposex. A comparação entre as variáveis morfométricas mostrou diferenças estatisticamente significantes entre os dois locais estudados. Além disso, as fêmeas da espécie S. haemastomaapresentaram uma frequência de imposex de 93,1% na Ilha das Aves, Baía das Cartagena, em comparação com 31,8% em La Bocana. O índice do comprimento relativo do pênis ou RPLI (10,145 e 3,231) e o índice da sequência do vaso deferente ou VDSI (2,83 e 1,16), mostraram possível contaminação por compostos organoestânicos em ambos os locais.

Effect of three polychlorinated biphenyls on f-met-leu-phe-induced degranulation in rat neutrophils.

It has been known that polychlorinated biphenyl (PCB) mixtures and individual congeners produce degranulation of rat neutrophils. Structure-activity relationships for congeners PCB 8 (2,4'-dichlorobiphenyl), PCB 126 (3,3',4,4',5-pentachlorobiphenyl) and PCB 128 (2,2',3,3',4,4'-hexachlorobiphenyl) were examined by correlating the extent of degranulation and cytotoxicity with molecular and physico-chemical parameters. Neutrophils were exposed to PCB congeners and then to the neutrophil activator f-met-leu-phe (fmlp). Degranulation and cytotoxicity were quantified by measuring released myeloperoxidase and lactate dehydrogenase activities, respectively. Degranulation in the absence of fmlp, that is in quiescent neutrophils, was detected only for PCB 8. Inhibition of fmlp-induced degranulation was observed for both PCB 8 (50 microM) and PCB 128 (10 and 50 microM). PCB 126 did not affect degranulation of quiescent or fmlp-stimulated neutrophils. Thus, effects on degranulation were observed only for ortho-substituted congeners. Cytotoxicity was observed under all conditions with PCB 8, in quiescent neutrophils with PCB 128, and in activated neutrophils with PCB 126. Structure-activity relationships revealed that effects of PCBs on neutrophil degranulation correlate with the energy of the lowest unoccupied molecular orbital but not with torsional angle 2,1,1',2'. This study demonstrates the importance of molecular, electronic parameters in PCB-induced effects on neutrophil degranulation.

Neutrophil activation by polychlorinated biphenyls: structure-activity relationship.

Polychlorinated biphenyls (PCBs) rapidly stimulate polymorphonuclear leukocytes (neutrophils) in vitro to produce superoxide anion (O2-). This response results from activation of various intracellular signal transduction pathways and appears to occur in a structure-specific fashion. Individual PCB congeners, varying in pattern and extent of chlorination, were tested for their ability to stimulate production of O2- and/or to enhance the response to protein kinase C activation by phorbol myristate acetate (PMA). Neutrophils were isolated from retired breeder, male, Sprague-Dawley rats and exposed to either vehicle, 10 or 50 microM PCB for 30 min at 37 degrees C. PMA (0 or 20 ng/ml) was added for an additional 10 min, and O2- generated during the incubation period was measured. 2,2'-Dichlorobiphenyl (2,2'-DCB), 2,4'-DCB, or 3,3'-DCB (50 microM) stimulated neutrophils to produce O2-. Incubation of neutrophils with 4,4'-DCB, 3,3',4,4',5-pentachlorobiphenyl (3,3',4,4',5-PeCB), 3,3',4,5,5'-PeCB, or 2,2',3,3',4,4'-hexachlorobiphenyl (2,2',3,3',4,4'-HCB) did not result in generation of O2-. Of the various congeners, 2,4'-DCB elicited the greatest production of O2-. Exposure to 10 microM 2,2'-DCB, 2,4'-DCB, 3,3'-DCB, or 2,2',3,3',4,4'-HCB prior to addition of PMA caused a significant increase in the amount of O2- produced, greater than that seen with either compound alone. PMA-stimulated O2- production was unaffected by prior exposure to 4,4'-DCB, 3,3',4,4',5-PeCB, or 3,3',4,5,5'-PeCB. In separate experiments, 3,3',4,4',5-PeCB inhibited the amount of O2- produced in response to activation with either 3,3'-DCB or 2,4'-DCB. Thus, it appears that congeners which are noncoplanar are capable of stimulating neutrophil O2- production. Coplanar congeners with high affinity for the Ah receptor do not activate neutrophils to produce O2- and may inhibit this response. These results are consistent with the hypothesis that PCBs stimulate neutrophil O2- production by a mechanism that is structure-specific and dependent on the chlorine substitution pattern of the biphenyl rings. Molecular modeling suggested that the sum of atomic charges on chlorine atoms is the most important descriptor for congeners which stimulate O2- production. The angle of rotation and the difference in energy between the highest occupied molecular orbital and the lowest unoccupied molecular orbital are integrative descriptors which, along with the sum of chlorine atomic charges, are associated with this biological activity.