Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 62
Filtrar
Más filtros

Banco de datos
País/Región como asunto
Tipo del documento
Intervalo de año de publicación
1.
Part Fibre Toxicol ; 18(1): 44, 2021 12 15.
Artículo en Inglés | MEDLINE | ID: mdl-34911549

RESUMEN

BACKGROUND: Air pollution is a complex mixture of particles and gases, yet current regulations are based on single toxicant levels failing to consider potential interactive outcomes of co-exposures. We examined transcriptomic changes after inhalation co-exposure to a particulate and a gaseous component of air pollution and hypothesized that co-exposure would induce significantly greater impairments to mitochondrial bioenergetics. A whole-body inhalation exposure to ultrafine carbon black (CB), and ozone (O3) was performed, and the impact of single and multiple exposures was studied at relevant deposition levels. C57BL/6 mice were exposed to CB (10 mg/m3) and/or O3 (2 ppm) for 3 h (either a single exposure or four independent exposures). RNA was isolated from lungs and mRNA sequencing performed using the Illumina HiSeq. Lung pathology was evaluated by histology and immunohistochemistry. Electron transport chain (ETC) activities, electron flow, hydrogen peroxide production, and ATP content were assessed. RESULTS: Compared to individual exposure groups, co-exposure induced significantly greater neutrophils and protein levels in broncho-alveolar lavage fluid as well as a significant increase in mRNA expression of oxidative stress and inflammation related genes. Similarly, a significant increase in hydrogen peroxide production was observed after co-exposure. After single and four exposures, co-exposure revealed a greater number of differentially expressed genes (2251 and 4072, respectively). Of these genes, 1188 (single exposure) and 2061 (four exposures) were uniquely differentially expressed, with 35 mitochondrial ETC mRNA transcripts significantly impacted after four exposures. Both O3 and co-exposure treatment significantly reduced ETC maximal activity for complexes I (- 39.3% and - 36.2%, respectively) and IV (- 55.1% and - 57.1%, respectively). Only co-exposure reduced ATP Synthase activity (- 35.7%) and total ATP content (30%). Further, the ability for ATP Synthase to function is limited by reduced electron flow (- 25%) and translation of subunits, such as ATP5F1, following co-exposure. CONCLUSIONS: CB and O3 co-exposure cause unique transcriptomic changes in the lungs that are characterized by functional deficits to mitochondrial bioenergetics. Alterations to ATP Synthase function and mitochondrial electron flow underly a pathological adaptation to lung injury induced by co-exposure.


Asunto(s)
Contaminantes Atmosféricos , Ozono , Contaminantes Atmosféricos/toxicidad , Animales , Exposición por Inhalación/efectos adversos , Pulmón , Ratones , Ratones Endogámicos C57BL , Mitocondrias , Ozono/toxicidad , Hollín/toxicidad , Transcriptoma
2.
Inhal Toxicol ; 32(1): 24-38, 2020 01.
Artículo en Inglés | MEDLINE | ID: mdl-32028803

RESUMEN

Objective: In this study, we compared in vitro and in vivo bioactivity of nitrogen-doped multi-walled carbon nanotubes (NDMWCNT) to MWCNT to test the hypothesis that nitrogen doping would alter bioactivity.Materials and Methods: High-resolution transmission electron microscopy (TEM) confirmed the multilayer structure of MWCNT with an average layer distance of 0.36 nm, which was not altered by nitrogen doping: the nanomaterials had similar widths and lengths. In vitro studies with THP-1 cells and alveolar macrophages from C57BL/6 mice demonstrated that NDMWCNT were less cytotoxic and stimulated less IL-1ß release compared to MWCNT. For in vivo studies, male C57BL/6J mice received a single dose of dispersion medium (DM), 2.5, 10 or 40 µg/mouse of NDMWCNT, or 40 µg/mouse of MWCNT by oropharyngeal aspiration. Animals were euthanized between 1 and 7 days post-exposure for whole lung lavage (WLL) studies.Results and Discussion: NDMWCNT caused time- and dose-dependent pulmonary inflammation. However, it was less than that caused by MWCNT. Activation of the NLRP3 inflammasome was assessed in particle-exposed mice by determining cytokine production in WLL fluid at 1 day post-exposure. Compared to DM-exposed mice, IL-1ß and IL-18 were significantly increased in MWCNT- and NDMWCNT-exposed mice, but the increase caused by NDMWCNT was less than MWCNT. At 56 days post-exposure, histopathology determined lung fibrosis in MWCNT-exposed mice was greater than NDMWCNT-exposed mice.Conclusions: These data indicate nitrogen doping of MWCNT decreases their bioactivity, as reflected with lower in vitro and in vivo toxicity inflammation and lung disease. The lower activation of the NLRP3 inflammasome may be responsible. Abbreviations: NDMWCNT: nitrogen-doped multi-walled carbon nanotubes; MWCNT: multi-walled carbon nanotubes; TEM: transmission electron microscopy; HRTEM: high resolution transmission electron microscopy; IL-1ß: interleukin-1ß; DM: dispersion medium; WLL: whole lung lavage; IL-18: interleukin-18; GSD: geometric standard deviation; XPS: X-ray photoelectron spectroscopy; SEM: standard error of the mean; PMA: phorbol 12-myristate 13-acetate; LPS: lipopolysacharride; LDH: lactate dehydrogenase; AM: alveolar macrophage; PMN: polymorphonuclear leukocyte.


Asunto(s)
Exposición por Inhalación/efectos adversos , Pulmón/efectos de los fármacos , Macrófagos Alveolares/efectos de los fármacos , Nanotubos de Carbono/toxicidad , Nitrógeno/toxicidad , Neumonía/inducido químicamente , Animales , Líquido del Lavado Bronquioalveolar/química , Citocinas/análisis , Relación Dosis-Respuesta a Droga , Humanos , Inflamasomas/inmunología , Inflamasomas/metabolismo , Pulmón/inmunología , Pulmón/patología , Macrófagos Alveolares/inmunología , Macrófagos Alveolares/patología , Masculino , Ratones Endogámicos C57BL , Nanotubos de Carbono/química , Nitrógeno/química , Tamaño de la Partícula , Neumonía/inmunología , Neumonía/patología , Propiedades de Superficie , Células THP-1 , Factores de Tiempo
3.
Part Fibre Toxicol ; 11: 3, 2014 Jan 09.
Artículo en Inglés | MEDLINE | ID: mdl-24405760

RESUMEN

BACKGROUND: Engineered carbon nanotubes are currently used in many consumer and industrial products such as paints, sunscreens, cosmetics, toiletries, electronic processes and industrial lubricants. Carbon nanotubes are among the more widely used nanoparticles and come in two major commercial forms, single-walled carbon nanotubes (SWCNT) and the more rigid, multi-walled carbon nanotubes (MWCNT). The low density and small size of these particles makes respiratory exposures likely. Many of the potential health hazards have not been investigated, including their potential for carcinogenicity. We, therefore, utilized a two stage initiation/promotion protocol to determine whether inhaled MWCNT act as a complete carcinogen and/or promote the growth of cells with existing DNA damage. Six week old, male, B6C3F1 mice received a single intraperitoneal (ip) injection of either the initiator methylcholanthrene(MCA, 10 µg/g BW, i.p.), or vehicle (corn oil). One week after i.p. injections, mice were exposed by inhalation to MWCNT (5 mg/m³, 5 hours/day, 5 days/week) or filtered air (controls) for a total of 15 days. At 17 months post-exposure, mice were euthanized and examined for lung tumor formation. RESULTS: Twenty-three percent of the filtered air controls, 26.5% of the MWCNT-exposed, and 51.9% of the MCA-exposed mice, had lung bronchiolo-alveolar adenomas and lung adenocarcinomas. The average number of tumors per mouse was 0.25, 0.81 and 0.38 respectively. By contrast, 90.5% of the mice which received MCA followed by MWCNT had bronchiolo-alveolar adenomas and adenocarcinomas with an average of 2.9 tumors per mouse 17 months after exposure. Indeed, 62% of the mice exposed to MCA followed by MWCNT had bronchiolo-alveolar adenocarcinomas compared to 13% of the mice that received filtered air, 22% of the MCA-exposed, or 14% of the MWCNT-exposed. Mice with early morbidity resulting in euthanasia had the highest rate of metastatic disease. Three mice exposed to both MCA and MWCNT that were euthanized early had lung adenocarcinoma with evidence of metastasis (5.5%). Five mice (9%) exposed to MCA and MWCNT and 1 (1.6%) exposed to MCA developed serosal tumors morphologically consistent with sarcomatous mesotheliomas, whereas mice administered MWCNT or air alone did not develop similar neoplasms. CONCLUSIONS: These data demonstrate that some MWCNT exposures promote the growth and neoplastic progression of initiated lung cells in B6C3F1 mice. In this study, the mouse MWCNT lung burden of 31.2 µg/mouse approximates feasible human occupational exposures. Therefore, the results of this study indicate that caution should be used to limit human exposures to MWCNT.


Asunto(s)
Adenocarcinoma/inducido químicamente , Neoplasias Pulmonares/inducido químicamente , Nanotubos de Carbono/toxicidad , Adenocarcinoma/patología , Adenocarcinoma del Pulmón , Adenoma/inducido químicamente , Adenoma/patología , Animales , Líquido del Lavado Bronquioalveolar/citología , Técnica del Anticuerpo Fluorescente , Hiperplasia/inducido químicamente , Hiperplasia/patología , Exposición por Inhalación , Pulmón/patología , Neoplasias Pulmonares/patología , Mesotelioma/inducido químicamente , Mesotelioma/patología , Ratones , Ratones Endogámicos , Microscopía de Polarización , Infiltración Neutrófila/efectos de los fármacos , Análisis de Supervivencia
4.
Redox Biol ; 46: 102092, 2021 10.
Artículo en Inglés | MEDLINE | ID: mdl-34418598

RESUMEN

Environmental inhalation exposures are inherently mixed (gases and particles), yet regulations are still based on single toxicant exposures. While the impacts of individual components of environmental pollution have received substantial attention, the impact of inhalation co-exposures is poorly understood. Here, we mechanistically investigated pulmonary inflammation and lung function decline after inhalation co-exposure and individual exposures to ozone (O3) and ultrafine carbon black (CB). Environmentally/occupationally relevant lung deposition levels in mice were achieved after inhalation of stable aerosols with similar aerodynamic and mass median distributions. X-ray photoemission spectroscopy detected increased surface oxygen contents on particles in co-exposure aerosols. Compared with individual exposures, co-exposure aerosols produced greater acellular and cellular oxidants detected by electron paramagnetic resonance (EPR) spectroscopy, and in vivo immune-spin trapping (IST), as well as synergistically increased lavage neutrophils, lavage proteins and inflammation related gene/protein expression. Co-exposure induced a significantly greater respiratory function decline compared to individual exposure. A synthetic catalase-superoxide dismutase mimetic (EUK-134) significantly blunted lung inflammation and respiratory function decline confirming the role of oxidant imbalance. We identified a significant induction of epithelial alarmin (thymic stromal lymphopoietin-TSLP)-dependent interleukin-13 pathway after co-exposure, associated with increased mucin and interferon gene expression. We provided evidence of interactive outcomes after air pollution constituent co-exposure and identified a key mechanistic pathway that can potentially explain epidemiological observation of lung function decline after an acute peak of air pollution. Developing and studying the co-exposure scenario in a standardized and controlled fashion will enable a better mechanistic understanding of how environmental exposures result in adverse outcomes.


Asunto(s)
Contaminantes Atmosféricos , Ozono , Neumonía , Contaminantes Atmosféricos/toxicidad , Alarminas/farmacología , Animales , Carbono/farmacología , Exposición por Inhalación , Pulmón , Ratones , Oxidantes/farmacología , Ozono/toxicidad , Tamaño de la Partícula , Neumonía/inducido químicamente
5.
Am J Respir Cell Mol Biol ; 43(2): 210-9, 2010 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-19783790

RESUMEN

DNA double-strand breaks (DSBs) can result in cell death or genetic alterations when cells are subjected to radiation, exposure to toxins, or other environmental stresses. A complex DNA-damage-response pathway is activated to repair the damage, and the inability to repair these breaks can lead to carcinogenesis. One of the earliest responses to DNA DSBs is the phosphorylation of a histone, H2AX, at serine 139 (gamma-H2AX), which can be detected by a fluorescent antibody. A study was undertaken to compare the induction of DNA DSBs in normal (small airway epithelial) cells and cancer cells (A549) after exposure to asbestos (crocidolite), a proven carcinogen, silica, a suspected carcinogen, and titanium dioxide (TiO(2)), an inert particle recently reported to be carcinogenic in animals. The results indicate that crocidolite induced greater DNA DSBs than silica and TiO(2), regardless of cell type. DNA DSBs caused by crocidolite were higher in normal cells than in cancer cells. Silica and TiO(2) induced higher DNA DSBs in cancer cells than in normal cells. The production of reactive oxygen species was found to be highest in cells exposed to crocidolite, followed, in potency, by silica and TiO(2). The generation of reactive oxygen species was higher in normal cells than in cancer cells. Cell viability assay indicated that crocidolite caused the greatest cytotoxicity in both cell types. Apoptosis, measured by caspase 3/7 and poly (ADP-Ribose) polymerase activation, was highest in crocidolite-exposed cells, followed by TiO(2) and silica. The results of this study indicate that crocidolite has a greater carcinogenic potential than silica and TiO(2), judged by its ability to cause sustained genomic instability in normal lung cells.


Asunto(s)
Amianto/farmacología , Biomarcadores de Tumor/metabolismo , Roturas del ADN de Doble Cadena , ADN/efectos de los fármacos , Neoplasias/inducido químicamente , Neoplasias/metabolismo , Dióxido de Silicio/farmacología , Titanio/farmacología , Carcinógenos/metabolismo , Caspasas/metabolismo , Línea Celular Tumoral , Supervivencia Celular , Espectroscopía de Resonancia por Spin del Electrón , Activación Enzimática , Humanos , Especies Reactivas de Oxígeno/metabolismo
6.
Biochim Biophys Acta ; 1796(2): 242-51, 2009 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19591900

RESUMEN

Hepatocellular carcinoma (HCC) is the third leading cause of cancer death worldwide. Despite tremendous efforts to diagnose and institute new treatment regimens, the prognosis is still extremely poor. Therefore, knowledge of the molecular mechanisms governing the initiation, maintenance and progression of HCC is urgently needed. Recently, several groups have attributed an important role for c-Jun N-terminal kinase 1 (JNK1) in the pathogenesis of human HCC and its close association with the expression of HCC signature genes. In this review the various associations between JNK1 and HCC are discussed with the hope that targeting this pivotal kinase may lead to novel therapeutic approaches for this fatal disease.


Asunto(s)
Carcinoma Hepatocelular/etiología , Neoplasias Hepáticas/etiología , Proteína Quinasa 8 Activada por Mitógenos/fisiología , Secuencia de Aminoácidos , Animales , Carcinoma Hepatocelular/tratamiento farmacológico , Carcinoma Hepatocelular/enzimología , Activación Enzimática , Humanos , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Hepáticas/enzimología , Proteína Quinasa 8 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 9 Activada por Mitógenos/fisiología , Datos de Secuencia Molecular , Especies Reactivas de Oxígeno/metabolismo , Ubiquitina/metabolismo
7.
Mol Cancer ; 9: 134, 2010 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-20515486

RESUMEN

MicroRNAs (miRNAs) are a class of small, noncoding RNAs critically involved in a wide spectrum of normal and pathological processes of cells or tissues by fine-tuning the signals important for stem cell development, cell differentiation, cell cycle regulation, apoptosis, and transformation. Considerable progress has been made in the past few years in understanding the transcription, biogenesis and functional regulation of miRNAs. Numerous studies have implicated altered expression of miRNAs in human cancers, suggesting that aberrant expression of miRNAs is one of the hallmarks for carcinogenesis. In this review, we briefly discuss most recent discoveries on the regulation of miRNAs at the level of microprocessor-mediated biogenesis of miRNAs.


Asunto(s)
Regulación de la Expresión Génica , MicroARNs/fisiología , Neoplasias/genética , Animales , Humanos
8.
Nat Mater ; 8(7): 543-57, 2009 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-19525947

RESUMEN

Rapid growth in nanotechnology is increasing the likelihood of engineered nanomaterials coming into contact with humans and the environment. Nanoparticles interacting with proteins, membranes, cells, DNA and organelles establish a series of nanoparticle/biological interfaces that depend on colloidal forces as well as dynamic biophysicochemical interactions. These interactions lead to the formation of protein coronas, particle wrapping, intracellular uptake and biocatalytic processes that could have biocompatible or bioadverse outcomes. For their part, the biomolecules may induce phase transformations, free energy releases, restructuring and dissolution at the nanomaterial surface. Probing these various interfaces allows the development of predictive relationships between structure and activity that are determined by nanomaterial properties such as size, shape, surface chemistry, roughness and surface coatings. This knowledge is important from the perspective of safe use of nanomaterials.


Asunto(s)
Nanopartículas/química , Materiales Biocompatibles/química , Materiales Biocompatibles/metabolismo , Modelos Biológicos , Nanotecnología/métodos , Nanotecnología/tendencias , Tamaño de la Partícula , Proteínas/química , Proteínas/metabolismo
9.
J Toxicol Environ Health A ; 73(5): 378-95, 2010.
Artículo en Inglés | MEDLINE | ID: mdl-20155580

RESUMEN

Carbon nanotubes (CNT), since their discovery, have become one of the most promising nanomaterials in many industrial and biomedical applications. Due to their unique physicochemical properties, interest is growing in the manufacture of CNT-based products and their subsequent marketing. Since their discovery, the prospect of possible undesirable human health effects has been a focus of many scientific studies. Although CNT possess unique physical properties that include (1) nanoscale diameter, (2) a wide length distribution ranging from tens of nanometers to several micrometers, and (3) high aspect ratio, the fibrous-like shape and durability suggest that their toxic properties may be analogous to those observed with other fibrous particles, such as asbestos. The present study provides a summary of published findings on CNT bioactivity, such as the potential of CNT, especially of multi-wall carbon nanotubes (MWCNT), to activate signaling pathways modulating transcription factor activity, induce apoptosis, induce DNA damage, and initiate biological responses. Assessment of risks to human health and adoption of appropriate exposure controls is critical for the safe and successful introduction of CNT -based products for future applications.


Asunto(s)
Amianto/toxicidad , Carcinógenos/toxicidad , Nanotubos de Carbono/toxicidad , Amianto/química , Carcinógenos/química , Carcinoma Broncogénico/inducido químicamente , Daño del ADN , Epitelio/metabolismo , Humanos , Mesotelioma/inducido químicamente , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Nanotubos de Carbono/química , Tamaño de la Partícula , Medición de Riesgo , Transducción de Señal/efectos de los fármacos , Factor de Transcripción AP-1/metabolismo
10.
Mol Cancer ; 8: 64, 2009 Aug 17.
Artículo en Inglés | MEDLINE | ID: mdl-19686584

RESUMEN

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide with an extremely poor prognosis. The classification of HCC based on the molecular signature is not well-established. RESULTS: In the present study, we reported HCC signature genes based on the JNK1 activation status in 31 HCC specimens relative to the matched distal noncancerous liver tissue from 31 patients. The HCCs with high JNK1 (H-JNK1) and low JNK1 (L-JNK1) were sub-grouped. Two different signature gene sets for both H-JNK1 and L-JNK1 HCC were identified through gene expression profiling. A striking overlap of signature genes was observed between the H-JNK1 HCC and the hepatoblastoma or hepatoblastoma-type HCC. Many established biomarkers for hepatic progenitor cells were over-expressed in H-JNK1 HCC, including AFP, TACSTD1, KRT19, KRT7, THY1, and PROM1. In addition, the majority of the most up-regulated genes were those associated with metastasis and earlier recurrence, whereas the genes for normal liver function were substantially down-regulated in H-JNK1 HCC tissue. A Kaplan-Meier plot demonstrated that the survival of the patients with H-JNK1 HCC was severely impaired. CONCLUSION: Accordingly, we believe that the H-JNK1 HCC may originate from hepatic progenitor cells and is associated with poorer prognosis. The status of JNK1 activation in HCC tissue, thus, might be a new biomarker for HCC prognosis and therapeutic targeting.


Asunto(s)
Carcinoma Hepatocelular/enzimología , Neoplasias Hepáticas/enzimología , Proteína Quinasa 8 Activada por Mitógenos/metabolismo , Adulto , Anciano , Carcinoma Hepatocelular/genética , Activación Enzimática , Femenino , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Estimación de Kaplan-Meier , Neoplasias Hepáticas/genética , Masculino , Persona de Mediana Edad , Proteína Quinasa 8 Activada por Mitógenos/genética , Pronóstico , Transducción de Señal , Análisis de Matrices Tisulares
11.
Mol Carcinog ; 48(5): 454-64, 2009 May.
Artículo en Inglés | MEDLINE | ID: mdl-18942077

RESUMEN

A new GADD45alpha isoform, GADD45alpha1, was identified in the cellular response to arsenic. DNA sequencing and biochemical analyses suggested that GADD45alpha1 is derived from an alternative splicing of the GADD45alpha mRNA by skipping the region corresponding to exon2 of the gadd45alpha gene during mRNA maturation. In addition to the size difference due to the lack of 34 amino acids encoded by exon2, GADD45alpha1 and GADD45alpha proteins differ in their effects on cell proliferation and cell cycle transition. Unlike GADD45alpha, the GADD45alpha1 is unable to attenuate cell growth. In over-expression experiments, the full length GADD45alpha, but not the GADD45alpha1, sensitized cells to arsenic-induced prometaphase arrest of the cell cycle. Furthermore, GADD45alpha1 appears to be able to antagonize the function of the GADD45alpha on the G2/M phase cell cycle arrest as demonstrated in cotransfection experiment. Thus, these data suggest that the generation of the GADD45alpha1 isoform may not only offset but also antagonize the effects of arsenic and GADD45alpha on cell growth and cell cycle regulation.


Asunto(s)
Empalme Alternativo , Arsénico/farmacología , Proteínas de Ciclo Celular/genética , Células Epiteliales/efectos de los fármacos , Proteínas Nucleares/genética , Isoformas de Proteínas/genética , Secuencia de Aminoácidos , Secuencia de Bases , Western Blotting , Ciclo Celular , Proteínas de Ciclo Celular/química , Proliferación Celular , Cartilla de ADN , Células Epiteliales/metabolismo , Técnica del Anticuerpo Fluorescente , Humanos , Datos de Secuencia Molecular , Proteínas Nucleares/química , Fosforilación , Isoformas de Proteínas/química , ARN Mensajero/genética
12.
Part Fibre Toxicol ; 6: 28, 2009 Oct 28.
Artículo en Inglés | MEDLINE | ID: mdl-19863807

RESUMEN

Particle and Fibre Toxicology wants to play a decisive role in a time where particle research is challenged and driven by the developments and applications of nanomaterials. This aim is not merely quantitative in publishing a given number of papers on nanomaterials, but also qualitatively since the field of nanotoxicology is rapidly emerging and benchmarks for good science are needed. Since then a number of things have happened that merit further analysis. The interactive learning issue is best shown by report and communications on the toxicology of multi-wall carbon nanotubes (CNT). A special workshop on the CNT has now been organized twice in Nagano (Japan) and this editorial contains a summary of the most important outcomes. Finally, we take the opportunity discuss some recent reports from the nanotech literature, and more specifically a Chinese study that claims severe consequences of nanoparticle exposure.

13.
J Toxicol Environ Health A ; 72(23): 1509-19, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-20077225

RESUMEN

Crystalline silica (silica), a suspected human carcinogen, produces an increase in reactive oxygen species (ROS) when fractured using mechanical tools used in several occupations. Although ROS has been linked to apoptosis, DNA damage, and carcinogenesis, the role of enhanced ROS production by silica in silica-induced carcinogenesis is not completely understood. The goal of this study was to compare freshly fractured and aged silica-induced molecular alterations in human immortalized/transformed bronchial epithelial cells (BEAS-IIB) and lung cancer cells with altered (H460) or deficient (H1299) p53 expression. Exposure to freshly fractured or aged silica produced divergent cellular responses in certain downstream cellular events, including ROS production, apoptosis, cell cycle and chromosomal changes, and gene expression. ROS production increased significantly following exposure to freshly fractured silica compared to aged silica in BEAS-IIB and H460 cells. Apoptosis showed a comparable enhanced level of induction with freshly fractured or aged silica in both cancer lines with p53 functional changes. p53 protein was present in the BEAS-IIB and was absent in cancer cell lines after silica exposure. Exposure to freshly fractured silica also resulted in a rise in aneuploidy in cancer cells with a significantly greater increase in p53-deficient cells. Cytogenetic analysis demonstrated increased metaphase spreads, chromosome breakage, rearrangements, and endoreduplication in both cancer cells. These results suggest that altered and deficient p53 affects the cellular response to freshly fractured silica exposure, and thereby enhances susceptibility and augments cell proliferation and lung cancer development.


Asunto(s)
Dióxido de Silicio/toxicidad , Proteína p53 Supresora de Tumor/metabolismo , Apoptosis/efectos de los fármacos , Pruebas de Carcinogenicidad , Línea Celular , Análisis Citogenético , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Especies Reactivas de Oxígeno/metabolismo , Proteína p53 Supresora de Tumor/genética
14.
J Toxicol Environ Health A ; 72(8): 560-70, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-19267316

RESUMEN

Pulmonary responses to diesel exhaust particles (DEP) exposure are mediated through enhanced production of reactive oxygen species (ROS) and nitric oxide (NO) by alveolar macrophages (AM). The current study examined the differential roles of ROS and NO in DEP-induced lung injury using C57B/6J wild-type (WT) and inducible NO synthase knockout (iNOS KO) mice. Mice exposed by pharyngeal aspiration to DEP or carbon black particles (CB) (35 mg/kg) showed an inflammatory profile that included neutrophil infiltration, increased lactate dehydrogenase (LDH) activity, and elevated albumin content in bronchoalveolar lavage fluid (BALF) at 1, 3, and 7 d postexposure. The organic extract of DEP (DEPE) did not induce an inflammatory response. Comparing WT to iNOS KO mice, the results show that NO enhanced DEP-induced neutrophils infiltration and plasma albumin content in BALF and upregulated the production of the pro-inflammatory cytokine interleukin 12 (IL-12) by AM. DEP-exposed AM from iNOS KO mice displayed diminished production of IL-12 and, in response to ex vivo lipopolysaccharide (LPS) challenge, decreased production of IL-12 but increased production of IL-10 when compared to cells from WT mice. DEP, CB, but not DEPE, induced DNA damage and mitochondria dysfunction in AM, however, that is independent of cellular production of NO. These results demonstrate that DEP-induced immune/inflammatory responses in mice are regulated by both ROS- and NO-mediated pathways. NO did not affect ROS-mediated mitochondrial dysfunction and DNA damage but upregulated IL-12 and provided a counterbalance to the ROS-mediated adaptive stress response that downregulates IL-12 and upregulates IL-10.


Asunto(s)
Enfermedades Mitocondriales/inducido químicamente , Óxido Nítrico Sintasa de Tipo II/fisiología , Óxido Nítrico/toxicidad , Material Particulado/toxicidad , Neumonía/inducido químicamente , Especies Reactivas de Oxígeno/toxicidad , Emisiones de Vehículos/toxicidad , Animales , Disponibilidad Biológica , Líquido del Lavado Bronquioalveolar/citología , Células Cultivadas , Ensayo Cometa , Citocinas/metabolismo , Daño del ADN , L-Lactato Deshidrogenasa/metabolismo , Ratones , Ratones Noqueados , Microscopía Confocal , Microscopía Electrónica de Transmisión , Enfermedades Mitocondriales/patología , Óxido Nítrico Sintasa de Tipo II/genética , Material Particulado/farmacocinética , Neumonía/patología , Alveolos Pulmonares/patología
15.
Cancer Res ; 67(13): 6146-54, 2007 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-17616671

RESUMEN

We have previously shown that trivalent arsenic (arsenite, As(3+)) is able to induce GADD45 alpha expression in human bronchial epithelial cells through activation of c-Jun NH(2)-terminal kinase and nucleolin-dependent mRNA stabilization. In the present report, we show that As(3+) is capable of inducing translation of the GADD45 alpha protein through a cap-independent, or rather, an internal ribosome entry site (IRES)-dependent mechanism. In growth-arrested cells, As(3+) elevated the GADD45 alpha protein level in a dose- and time-dependent manner which did not correlate with the GADD45 alpha mRNA expression. Pretreatment of the cells with rapamycin, an inhibitor for the cap-dependent translation machinery through the suppression of mTOR and p70S6 kinase, failed to affect the induction of the GADD45 alpha protein induced by As(3+). Sequence analysis revealed a potential IRES element in the 5'-untranslated region of the GADD45 alpha mRNA. This IRES element in the 5'-untranslated region of the GADD45 alpha mRNA is functional in mediating As(3+)-induced translation of the GADD45 alpha protein in a dicistronic reporter gene activity assay. Immunoprecipitation and proteomic studies suggest that As(3+) impairs the assembly of the cap-dependent initiating complex for general protein translation but increases the association of human elongation factor 2 and human heterogeneous nuclear ribonucleoprotin with this complex. Thus, these results suggest that in growth-arrested cells, As(3+) is still capable of inducing GADD45 alpha expression through an IRES-dependent translational regulation.


Asunto(s)
Arsénico/farmacología , Proteínas de Ciclo Celular/biosíntesis , Regulación Neoplásica de la Expresión Génica , Proteínas Nucleares/biosíntesis , Ribosomas/genética , Secuencia de Aminoácidos , Secuencia de Bases , Proteínas de Ciclo Celular/genética , Genes Reporteros , Ribonucleoproteínas Nucleares Heterogéneas/metabolismo , Humanos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Datos de Secuencia Molecular , Proteínas Nucleares/genética , Regiones Promotoras Genéticas , ARN Mensajero/metabolismo , Proteínas Quinasas S6 Ribosómicas 70-kDa/metabolismo , Homología de Secuencia de Aminoácido
16.
Am J Respir Cell Mol Biol ; 38(5): 579-90, 2008 May.
Artículo en Inglés | MEDLINE | ID: mdl-18096873

RESUMEN

Carbon nanotubes (CNT), with their applications in industry and medicine, may lead to new risks to human health. CNT induce a robust pulmonary inflammation and oxidative stress in rodents. Realistic exposures to CNT may occur in conjunction with other pathogenic impacts (microbial infections) and trigger enhanced responses. We evaluated interactions between pharyngeal aspiration of single-walled CNT (SWCNT) and bacterial pulmonary infection of C57BL/6 mice with Listeria monocytogenes (LM). Mice were given SWCNT (0, 10, and 40 mug/mouse) and 3 days later were exposed to LM (10(3) bacteria/mouse). Sequential exposure to SWCNT/LM amplified lung inflammation and collagen formation. Despite this robust inflammatory response, SWCNT pre-exposure significantly decreased the pulmonary clearance of LM-exposed mice measured 3 to 7 days after microbial infection versus PBS/LM-treated mice. Decreased bacterial clearance in SWCNT-pre-exposed mice was associated with decreased phagocytosis of bacteria by macrophages and a decrease in nitric oxide production by these phagocytes. Pre-incubation of naïve alveolar macrophages with SWCNT in vitro also resulted in decreased nitric oxide generation and suppressed phagocytizing activity toward LM. Failure of SWCNT-exposed mice to clear LM led to a continued elevation in nearly all major chemokines and acute phase cytokines into the later course of infection. In SWCNT/LM-exposed mice, bronchoalveolar lavage neutrophils, alveolar macrophages, and lymphocytes, as well as lactate dehydrogenase level, were increased compared with mice exposed to SWCNT or LM alone. In conclusion, enhanced acute inflammation and pulmonary injury with delayed bacterial clearance after SWCNT exposure may lead to increased susceptibility to lung infection in exposed populations.


Asunto(s)
Listeria monocytogenes/patogenicidad , Listeriosis/patología , Pulmón/patología , Nanotubos de Carbono/microbiología , Neumonía/inducido químicamente , Neumonía/microbiología , Animales , Líquido del Lavado Bronquioalveolar/inmunología , Líquido del Lavado Bronquioalveolar/microbiología , Citocinas/biosíntesis , Femenino , Listeriosis/inmunología , Listeriosis/fisiopatología , Pulmón/inmunología , Pulmón/microbiología , Macrófagos Alveolares/inmunología , Ratones , Ratones Endogámicos C57BL , Nanotubos de Carbono/toxicidad , Fagocitosis , Neumonía/inmunología , Neumonía/patología , Pérdida de Peso
17.
Nucleic Acids Res ; 34(2): 485-95, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-16421274

RESUMEN

The present study shows that arsenic induces GADD45alpha (growth arrest and DNA damage inducible gene 45alpha) mainly through post-transcriptional mechanism. Treatment of the human bronchial epithelial cell line, BEAS-2B, with arsenic(III) chloride (As3+) resulted in a significant increase in GADD45alpha protein and mRNA. However, As3+ only exhibited a marginal effect on the transcription of the GADD45alpha gene. The accumulation of GADD45alpha mRNA is largely achieved by the stabilization of GADD45alpha mRNA in the cellular response to As3+. As3+ is able to induce binding of mRNA stabilizing proteins, nucleolin and less potently, HuR, to the GADD45alpha mRNA. Although As3+ was unable to affect the expression of nucleolin, treatment of the cells with As3+ resulted in re-distribution of nucleolin from nucleoli to nucleoplasm. Silencing of the nucleolin mRNA by RNA interference reversed As3+-induced stabilization of the GADD45alpha mRNA and accumulation of the GADD45alpha protein. Stabilization of GADD45alpha mRNA, thus, represents a novel mechanism contributing to the production of GADD45alpha and cell cycle arrest in response to As3+.


Asunto(s)
Arsenicales/farmacología , Proteínas de Ciclo Celular/genética , Cloruros/farmacología , Proteínas Nucleares/genética , Fosfoproteínas/metabolismo , Estabilidad del ARN/efectos de los fármacos , ARN Mensajero/metabolismo , Proteínas de Unión al ARN/metabolismo , Proteínas de Ciclo Celular/metabolismo , Línea Celular , Silenciador del Gen , Humanos , Proteínas Nucleares/metabolismo , Fosfoproteínas/antagonistas & inhibidores , Fosfoproteínas/genética , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , Proteínas de Unión al ARN/antagonistas & inhibidores , Proteínas de Unión al ARN/genética , Transcripción Genética/efectos de los fármacos , Nucleolina
18.
Environ Health Perspect ; 115(5): 756-63, 2007 May.
Artículo en Inglés | MEDLINE | ID: mdl-17520064

RESUMEN

BACKGROUND: Inhalation of diesel exhaust particles (DEPs) is characterized by lung injury and inflammation, with significant increases in the numbers of polymorphonuclear leukocytes and alveolar macrophages. This influx of cellular infiltrates is associated with the activation of multiple genes, including cytokines and chemokines, and the production of reactive oxygen species. OBJECTIVE: The pathogenesis of the lung injury is not fully understood, but alterations in the presence or abundance of a number of proteins in the lung have been observed. Our objective in this study was to further characterize these changes and to ask whether additional changes could be discerned using modern proteomic techniques. METHODS: The present study investigates global alterations in the proteome of bronchoalveolar lavage fluid taken from rats 1, 7, or 30 days after exposure to 5, 35, or 50 mg/kg of animal weight of DEPs. RESULTS: Analysis by surface-enhanced laser desorption/ionization-time of flight mass spectrometry identified two distinct peaks that appeared as an acute response postexposure at all doses in all animals. We identified these two peaks, with mass to charge ratios (m/z) of 9,100 and 10,100, as anaphylatoxin C3a and calgranulin A by additional mass spectral investigation using liquid chromatography coupled to mass spectrometry. CONCLUSIONS: With this approach, we found a number of inflammatory response proteins that may be associated with the early phases of inflammation in response to DEP exposure. Further studies are warranted to determine whether serum levels of these proteins could be markers of diesel exhaust exposure in workers.


Asunto(s)
Líquido del Lavado Bronquioalveolar/química , Regulación de la Expresión Génica/efectos de los fármacos , Pulmón/efectos de los fármacos , Proteínas/análisis , Emisiones de Vehículos/toxicidad , Animales , Cromatografía Líquida de Alta Presión , Relación Dosis-Respuesta a Droga , Pulmón/metabolismo , Proteómica/métodos , Ratas , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Factores de Tiempo , Emisiones de Vehículos/análisis
19.
Toxicology ; 236(1-2): 103-13, 2007 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-17482744

RESUMEN

Firefighting, along with construction, mining and agriculture, ranks among the most dangerous occupations. In addition, the work environment of firefighters is unlike that of any other occupation, not only because of the obvious physical hazards but also due to the respiratory and systemic health hazards of smoke inhalation resulting from combustion. A significant amount of research has been devoted to studying municipal firefighters; however, these studies may not be useful in wildland firefighter exposures, because the two work environments are so different. Not only are wildland firefighters exposed to different combustion products, but their exposure profiles are different. The combustion products wildland firefighters are exposed to can vary greatly in characteristics due to the type and amount of material being burned, soil conditions, temperature and exposure time. Smoke inhalation is one of the greatest concerns for firefighter health and it has been shown that the smoke consists of a large number of particles. These smoke particles contain intermediates of hydrogen, carbon and oxygen free radicals, which may pose a potential health risk. Our investigation looked into the involvement of free radicals in smoke toxicity and the relationship between particle size and radical generation. Samples were collected in discrete aerodynamic particle sizes from a wildfire in Alaska, preserved and then shipped to our laboratory for analysis. Electron spin resonance was used to measure carbon-centered as well as hydroxyl radicals produced by a Fenton-like reaction with wildfire smoke. Further study of reactive oxygen species was conducted using analysis of cellular H(2)O(2) generation, lipid peroxidation of cellular membranes and DNA damage. Results demonstrate that coarse size-range particles contained more carbon radicals per unit mass than the ultrafine particles; however, the ultrafine particles generated more *OH radicals in the acellular Fenton-like reaction. The ultrafine particles also caused significant increases in H(2)O(2) production by monocytes and lipid peroxidation. All particle sizes showed the ability to cause DNA damage. These results indicate that the radical generation and the damage caused by them is not only a function of surface area but is also influenced by changing chemical and other characteristics due to particle size.


Asunto(s)
Contaminantes Atmosféricos/toxicidad , Incendios , Radicales Libres/toxicidad , Humo/efectos adversos , Contaminantes Atmosféricos/análisis , Animales , Línea Celular , Daño del ADN , Radicales Libres/análisis , Peróxido de Hidrógeno/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Ratones , Monocitos/efectos de los fármacos , Monocitos/metabolismo , Tamaño de la Partícula , Humo/análisis
20.
Clin Cancer Res ; 12(11 Pt 1): 3344-54, 2006 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-16740756

RESUMEN

PURPOSE: Individualized therapy of lung adenocarcinoma depends on the accurate classification of patients into subgroups of poor and good prognosis, which reflects a different probability of disease recurrence and survival following therapy. However, it is currently impossible to reliably identify specific high-risk patients. Here, we propose a computational model system which accurately predicts the clinical outcome of individual patients based on their gene expression profiles. EXPERIMENTAL DESIGN: Gene signatures were selected using feature selection algorithms random forests, correlation-based feature selection, and gain ratio attribute selection. Prediction models were built using random committee and Bayesian belief networks. The prognostic power of the survival predictors was also evaluated using hierarchical cluster analysis and Kaplan-Meier analysis. RESULTS: The predictive accuracy of an identified 37-gene survival signature is 0.96 as measured by the area under the time-dependent receiver operating curves. The cluster analysis, using the 37-gene signature, aggregates the patient samples into three groups with distinct prognoses (Kaplan-Meier analysis, P < 0.0005, log-rank test). All patients in cluster 1 were in stage I, with N0 lymph node status (no metastasis) and smaller tumor size (T1 or T2). Additionally, a 12-gene signature correctly predicts the stage of 94.2% of patients. CONCLUSIONS: Our results show that the prediction models based on the expression levels of a small number of marker genes could accurately predict patient outcome for individualized therapy of lung adenocarcinoma. Such an individualized treatment may significantly increase survival due to the optimization of treatment procedures and improve lung cancer survival every year through the 5-year checkpoint.


Asunto(s)
Adenocarcinoma/diagnóstico , Neoplasias Pulmonares/diagnóstico , Adenocarcinoma/clasificación , Adenocarcinoma/genética , Área Bajo la Curva , Análisis por Conglomerados , Simulación por Computador , Perfilación de la Expresión Génica , Humanos , Estimación de Kaplan-Meier , Neoplasias Pulmonares/clasificación , Neoplasias Pulmonares/genética , Valor Predictivo de las Pruebas , Pronóstico , Curva ROC , Análisis de Supervivencia , Factores de Tiempo
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA