RESUMO
Pharmacological induction of fetal hemoglobin (HbF) expression is an effective therapeutic strategy for the management of beta-hemoglobinopathies such as sickle cell disease. DNA methyltransferase (DNMT) inhibitors 5-azacytidine (5-aza) and 5-aza-2'-deoxycytidine (decitabine) have been shown to induce fetal hemoglobin expression in both preclinical models and clinical studies, but are not currently approved for the management of hemoglobinopathies. We report here the discovery of a novel class of orally bioavailable DNMT1-selective inhibitors as exemplified by GSK3482364. This molecule potently inhibits the methyltransferase activity of DNMT1, but not DNMT family members DNMT3A or DNMT3B. In contrast with cytidine analog DNMT inhibitors, the DNMT1 inhibitory mechanism of GSK3482364 does not require DNA incorporation and is reversible. In cultured human erythroid progenitor cells (EPCs), GSK3482364 decreased overall DNA methylation resulting in de-repression of the gamma globin genes HBG1 and HBG2 and increased HbF expression. In a transgenic mouse model of sickle cell disease, orally administered GSK3482364 caused significant increases in both HbF levels and in the percentage HbF-expressing erythrocytes, with good overall tolerability. We conclude that in these preclinical models, selective, reversible inhibition of DNMT1 is sufficient for the induction of HbF, and is well-tolerated. We anticipate that GSK3482364 will be a useful tool molecule for the further study of selective DNMT1 inhibition both in vitro and in vivo.
Assuntos
Anemia Falciforme , Hemoglobina Fetal , Anemia Falciforme/tratamento farmacológico , Anemia Falciforme/genética , Animais , Azacitidina/farmacologia , Metilação de DNA , Hemoglobina Fetal/genética , Camundongos , gama-Globinas/genéticaRESUMO
Data from toxicity tests of the pore water extracted from Puget Sound sediments were compiled from surveys conducted from 1997 to 2009. Tests were performed on 664 samples collected throughout all of the eight monitoring regions in the Sound, an area encompassing 2,294.1 km(2). Tests were performed with the gametes of the Pacific purple sea urchin, Strongylocentrotus purpuratus, to measure percent fertilization success as an indicator of relative sediment quality. Data were evaluated to determine the incidence, degree of response, geographic patterns, spatial extent, and temporal changes in toxicity. This is the first survey of this kind and magnitude in Puget Sound. In the initial round of surveys of the eight regions, 40 of 381 samples were toxic for an incidence of 10.5 %. Stations classified as toxic represented an estimated total of 107.1 km(2), equivalent to 4.7 % of the total area. Percent sea urchin fertilization ranged from >100 % of the nontoxic, negative controls to 0 %. Toxicity was most prevalent and pervasive in the industrialized harbors and lowest in the deep basins. Conditions were intermediate in deep-water passages, urban bays, and rural bays. A second round of testing in four regions and three selected urban bays was completed 5-10 years following the first round. The incidence and spatial extent of toxicity decreased in two of the regions and two of the bays and increased in the other two regions and the third bay; however, only the latter change was statistically significant. Both the incidence and spatial extent of toxicity were lower in the Sound than in most other US estuaries and marine bays.
Assuntos
Monitoramento Ambiental , Sedimentos Geológicos/química , Poluentes Químicos da Água/toxicidade , Análise Espaço-Temporal , Washington , Poluentes Químicos da Água/análise , Poluição Química da Água/estatística & dados numéricosRESUMO
The purpose of this work was to use various molecular imaging techniques to non-invasively assess GSK2849330 (anti HER3 ADCC and CDC enhanced 'AccretaMab' monoclonal antibody) pharmacokinetics and pharmacodynamics in human xenograft tumor-bearing mice. Immuno-PET biodistribution imaging of radiolabeled 89Zr-GSK2849330 was assessed in mice with HER3 negative (MIA-PaCa-2) and positive (CHL-1) human xenograft tumors. Dose dependency of GSK2849330 disposition was assessed using varying doses of unlabeled GSK2849330 co-injected with 89Zr-GSK2849330. In-vivo NIRF optical imaging and ex-vivo confocal microscopy were used to assess the biodistribution of GSK2849330 and the HER3 receptor occupancy in HER3 positive xenograft tumors (BxPC3, and CHL-1). Ferumoxytol (USPIO) contrast-enhanced MRI was used to investigate the effects of GSK2849330 on tumor macrophage content in CHL-1 xenograft bearing mice. Immuno-PET imaging was used to monitor the whole body drug biodistribution and CHL-1 xenograft tumor uptake up to 144 hours post injection of 89Zr-GSK2849330. Both hepatic and tumor uptake were dose dependent and saturable. The optical imaging data in the BxPC3 xenograft tumor confirmed the tumor dose response finding in the Immuno-PET study. Confocal microscopy showed a distinguished cytoplasmic punctate staining pattern within individual CHL-1 cells. GSK2849330 inhibited tumor growth and this was associated with a significant decrease in MRI signal to noise ratio after USPIO injection and with a significant increase in tumor macrophages as confirmed by a quantitative immunohistochemistry analysis. By providing both dose response and time course data from both 89Zr and fluorescently labeled GSK2849330, complementary imaging studies were used to characterize GSK2849330 biodistribution and tumor uptake in vivo. Ferumoxytol-enhanced MRI was used to monitor aspects of the immune system response to GSK2849330. Together these approaches potentially provide clinically translatable, non-invasive techniques to support dose optimization, and assess immune activation and anti-tumor responses.
Assuntos
Anticorpos Monoclonais Humanizados/farmacocinética , Anticorpos Monoclonais/farmacocinética , Macrófagos/imunologia , Compostos Radiofarmacêuticos/farmacocinética , Receptor ErbB-3/imunologia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais Humanizados/química , Anticorpos Monoclonais Humanizados/uso terapêutico , Linhagem Celular Tumoral , Feminino , Óxido Ferroso-Férrico/química , Humanos , Imuno-Histoquímica , Marcação por Isótopo , Macrófagos/citologia , Macrófagos/patologia , Camundongos , Camundongos Nus , Neoplasias/diagnóstico por imagem , Neoplasias/tratamento farmacológico , Radioisótopos , Compostos Radiofarmacêuticos/química , Compostos Radiofarmacêuticos/uso terapêutico , Receptor ErbB-3/metabolismo , Distribuição Tecidual , Transplante Heterólogo , Zircônio/químicaRESUMO
An internally consistent dataset comprising 103 surficial estuarine sediment samples were collected from Sydney Harbour, Australia and locations south of Sydney. This paper describes the chemical characteristics of the dataset and evaluates its suitability for use in evaluating biological effects-based sediment quality guidelines (SQGs). The sediments contained mixtures of chemicals, the most prevalent chemical classes being metals and polycyclic aromatic hydrocarbons, whereas sediments from coastal lakes/estuaries south of Sydney had low concentrations of contaminants. Maximum concentrations of the prevalent contaminants zinc, lead, copper and pyrene were 11,300, 1,420, 1,060 mg kg(-1) and 23,300 microg kg(-1), respectively. For the majority of samples, concentrations of individual chemicals exceeded most effects-based SQGs that have been adopted for use in Australia, implying occasional or frequent adverse biological effects are expected. Comparing mixtures of contaminants to ranges in numbers of SQGs exceeded and mean SQG quotients showed that most samples (57% to 68%) had contamination characteristics associated with moderate probabilities (30% to 52%) of acute toxicity, based on North American data. A smaller proportion of samples (15% to 17%) had contamination characteristics associated with high probabilities (74% to 85%) of toxicity. The wide range of chemicals and concentrations, associated with low, medium and high probabilities of toxicity, indicated that the dataset was suitable for future use in evaluating predictive abilities of SQGs. This is relevant, given the recent introduction of North American-derived SQGs for Australia.
Assuntos
Monitoramento Ambiental , Sedimentos Geológicos/análise , Metais Pesados/análise , Praguicidas/análise , Poluentes Químicos da Água/análise , Austrália , Sedimentos Geológicos/química , Guias como Assunto , Metais Pesados/toxicidade , Praguicidas/toxicidade , Bifenilos Policlorados/análise , Bifenilos Policlorados/toxicidade , Hidrocarbonetos Policíclicos Aromáticos/análise , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Medição de Risco , Poluentes Químicos da Água/toxicidadeRESUMO
Matching chemical and toxicological data of surficial sediments from Sydney Harbour, Australia, and vicinity, were collected to evaluate predictive abilities of Effects Range-Low (ERL), Effects Range-Median (ERM) and other sediment quality guidelines (SQGs). Samples (n=103) containing a wide range of chemicals and concentrations were subjected to a battery of 4-6 toxicity tests. ERLs and functionally equivalent low-range SQGs were highly predictive of non-toxicity when not exceeded, as incidences of toxicity were 0-8%. ERMs and other mid-range SQGs were predictive of toxicity in combined test data with > or = 80% of toxic samples with one or more SQG exceeded. Predictive abilities of mid-range SQGs were low for amphipod survival tests (generally < 20% toxic samples), but the incidence of toxicity increased with increasing numbers of SQGs exceeded and increasing mean SQG quotients. Predictive abilities of SQGs generally matched their narrative intent when outcomes of multiple toxicity tests were considered, and were consistent with North American data. Functionally equivalent SQGs gave comparable results (except where developed for single chemical classes), indicating that predictive abilities were primarily influenced by sensitivities of test species and/or the bioavailability of contaminants.
Assuntos
Monitoramento Ambiental , Sedimentos Geológicos/análise , Guias como Assunto , Testes de Toxicidade , Poluentes Químicos da Água/análise , Austrália , Monitoramento Ambiental/métodos , Monitoramento Ambiental/normas , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Testes de Toxicidade/métodos , Testes de Toxicidade/normasRESUMO
Recent studies by researchers at the U.S. Environmental Protection Agency and U.S. Geological Survey have evaluated the toxicity of Cr in freshwater and marine sediments, primarily during laboratory studies in which clean sediments were spiked with Cr. Results of those studies showed that Cr is relatively insoluble and nontoxic when present in the trivalent form, Cr(III), rather than in the more soluble and more toxic hexavalent form, Cr(VI). The studies concluded that Cr toxicity should be low in sediments with measurable concentrations of acid-volatile sulfide (AVS), because AVS is formed only in anoxic sediments and Cr(VI) is thermodynamically unstable under such conditions. The present study evaluates the toxicity and bioavailability of Cr in sediments associated with chromite ore processing residue (COPR). Ten stations were sampled in the Hackensack River (NJ, USA) to represent a wide range of total Cr concentrations (199-3,970 mg/kg) with minimal interference from potentially toxic, co-occurring chemicals. Sediment toxicity was evaluated using two amphipod tests: The 10-d Ampelisca abdita test (survival as endpoint), and the 28-d Leptocheirus plumulosus test (survival and biomass as endpoints). Measurable concentrations of AVS were present at eight stations, and nearly all Cr was present as Cr(III). In addition, results of electron-microprobe analyses showed that most Cr was associated with phases in which Cr has limited bioavailability (i.e., chromite and iron oxide). Sediment toxicity showed no correlation with concentrations of total Cr, and the maximum no-effect concentration for total Cr was estimated as 1,310 mg/ kg. These results indicate that Cr can be present in sediments associated with COPR at highly elevated concentrations without causing sediment toxicity.
Assuntos
Cromo/toxicidade , Sedimentos Geológicos/química , Disponibilidade Biológica , Cromo/farmacocinética , Microanálise por Sonda EletrônicaRESUMO
Laboratory survival tests were conducted with an indigenous infaunal amphipod, Corophium colo, on 103 sediment samples from Sydney Harbor (NSW, Australia) and vicinity, containing a wide range of chemicals and concentrations. The present study describes the sensitivity of C. colo to the sediments and compares the results to data for North American amphipods (Rhepoxynius abronius and Ampelisca abdita) previously used to establish and validate sediment-quality guidelines (SQGs). The incidence of toxicity increased with increasing contamination, as indicated by increasing numbers of SQGs exceeded and increasing mean SQG quotients. The incidence of highly toxic results (p < 0.05 and mean amphipod survival of < 80% that of controls) for highly contaminated samples was approximately half (28-40%) that of a large U.S. database (74%). The incidence of highly toxic responses for samples with intermediate levels of contamination also was lower in the present study (5-13%) compared to the results in large U.S. studies (approximately 30-50%). Corophium colo reburial tests showed greater sensitivity compared to survival tests, with a maximum incidence of statistically significant responses in moderately contaminated sediments of 70%. The present study showed that adult Corophium organisms are suitable for testing lethal responses in highly contaminated sediments (i.e., with mean effects range-median quotients of >1.5). Reburial results provide additional sensitivity.
Assuntos
Anfípodes , Sedimentos Geológicos/química , Poluentes da Água/toxicidade , Animais , Monitoramento Ambiental , New South Wales , Valores de Referência , Sobrevida , Testes de ToxicidadeRESUMO
Concentrations of 119 pharmaceuticals and personal care products (PPCPs) and 13 perfluoroalkyl substances (PFASs) in marine sediments measured throughout Puget Sound (n = 10) and Bellingham Bay (n = 30), Washington, USA, are reported. These data are among the first measurements of PPCPs and PFASs in marine sediments from the Pacific Northwest and provide a comparison to previous measurements of these chemicals in influent, effluent, and biosolids from municipal wastewater treatment plants throughout the region. The concentrations of both PPCPs and PFASs in sediments from Puget Sound and Bellingham Bay ranged from very low to non-detectable for most compounds. Only 14 of the 119 PPCPs and 3 of 13 PFASs were quantifiable in sediments. Diphenhydramine (an antihistamine) was most frequently detected (87.5% of samples), with a maximum concentration of 4.81 ng/g dry weight and an estimated mean detected concentration of 1.68 ng/g. Triclocarban (an antibacterial) was detected in 35.0% of the samples, with a maximum concentration of 16.6 ng/g dry weight. Perfluoroalkyl substances were detected in 2.5% of analyses. Perfluorobutanoate, perfluorooctane sulfonate, and perfluorooctane sulfonamide were detected in 7, 5, and 1 sample(s) each, respectively, with the highest concentrations observed for perfluorooctane sulfonate (1.5 ng/g). Detected concentrations were often highest within the industrial harbor in Bellingham Bay and near the cities of Seattle and Bremerton. Environ Toxicol Chem 2013;32:1701-1710. © 2013 SETAC.
Assuntos
Cosméticos/análise , Monitoramento Ambiental , Fluorocarbonos/análise , Sedimentos Geológicos/química , Preparações Farmacêuticas/análise , Poluentes Químicos da Água/análise , Ácidos Alcanossulfônicos/análise , Carbanilidas/análise , Cidades , Washington , Poluição Química da Água/estatística & dados numéricosRESUMO
The Washington State Department of Ecology annually conducts sediment quality monitoring in Puget Sound as a component of the Puget Sound Ecosystem Monitoring Program. Sediment samples are analyzed to determine the concentrations of about 170 chemical and physical variables. A Sediment Chemistry Index (SCI) was derived using the State of Washington Sediment Management Standards to account for the presence and concentrations of mixtures of toxicants. Mean Sediment Quality Standard quotients (mSQSq) were calculated as the basis for the SCI and compared to the incidence and degree of toxicity in laboratory tests and to metrics of the diversity and abundance of resident benthic assemblages in a database consisting of as many as 664 samples. These data were evaluated with co-occurrence analyses to identify "cut points" (i.e., thresholds) in the index below which the frequency and magnitude of biological effects were relatively low and above which they occurred with increasing frequency or magnitude. Iterative trials of different sets of cut points established the final cut points in mSQSq of 0.1, 0.3, and 0.5. They defined 4 ranges in chemical exposure: Minimum (<0.1), Low (0.1- < 0.3), Moderate (0.3- < 0.5), and Maximum (≥0.5). Across these 4 exposure ranges both the incidence and magnitude of toxicity in some laboratory tests increased, the abundance of most stress-sensitive benthic taxa decreased, and the abundance of most stress-tolerant taxa increased. The mSQSq cut point of 0.1 appears to be the target value for protection of benthic resources, the value below which the probability and magnitude of adverse effects either in the laboratory or the field are the lowest. The mSQSq values are rescaled from 0 to 100 to form the SCI, used by the Puget Sound Partnership and environmental managers as a Dashboard Indicator, with biologically relevant targets selected to monitor ecosystem recovery.
Assuntos
Baías/química , Ecotoxicologia/métodos , Monitoramento Ambiental/métodos , Poluentes Ambientais/análise , Sedimentos Geológicos/química , Ecotoxicologia/normas , Monitoramento Ambiental/normas , Poluentes Ambientais/química , Poluentes Ambientais/toxicidade , Laboratórios , Padrões de Referência , WashingtonRESUMO
The lower Passaic River in northern New Jersey (USA) has been heavily industrialized since the mid-nineteenth century and its shoreline and aquatic habitats degraded or destroyed. Similar to other urban systems, Passaic River sediments, both surface and buried, historically have contained elevated levels of numerous contaminants that may pose risks to ecological receptors and humans. Sediments from 15 stations in the lower Passaic River and 3 reference stations in the Mullica River in southern New Jersey were sampled in 1999 and characterized for chemical contamination, toxicity, and impairment of the benthic community. The objective of this study was to determine the incidence, degree, and nature of degraded surficial sediments in the area to support subsequent plans for restoration of the system. Results demonstrated that Passaic River sediments had concentrations of many organic and inorganic contaminants at levels significantly greater than the reference area and effect-based guidelines. Sediments were toxic to marine amphipods at 11 stations and the benthic assemblages were impaired relative to the reference area at all stations. The weight-of-evidence of this sediment quality triad (SQT) assessment indicates that impacts from multiple contaminants are occurring throughout the lower Passaic River and, that these impacts must be evaluated further and addressed as part of ongoing restoration initiatives for the river.
Assuntos
Água Doce/química , Sedimentos Geológicos/química , Poluentes Químicos da Água/análise , New England , Poluentes Químicos da Água/toxicidadeRESUMO
Fine-grained sediments contaminated with complex mixtures of organic and inorganic chemical contaminants can be toxic in laboratory tests and/or cause adverse impacts to resident benthic communities. Effects-based, sediment quality guidelines (SQGs) have been developed over the past 20 years to aid in the interpretation of the relationships between chemical contamination and measures of adverse biological effects. Mean sediment quality guideline quotients (mSQGQ) can be calculated by dividing the concentrations of chemicals in sediments by their respective SQGs and calculating the mean of the quotients for the individual chemicals. The resulting index provides a method of accounting for both the presence and the concentrations of multiple chemicals in sediments relative to their effects-based guidelines. Analyses of considerable amounts of data demonstrated that both the incidence and magnitude of toxicity in laboratory tests and the incidence of impairment to benthic communities increases incrementally with increasing mSQGQs. Such concentration/response relationships provide a basis for estimating toxicological risks to sediment-dwelling organisms associated with exposure to contaminated sediments with a known degree of accuracy. This sediment quality assessment tool has been used in numerous surveys and studies since 1994. Nevertheless, mean SQGQs have some important limitations and underlying assumptions that should be understood by sediment quality assessors. This paper provides an overview of the derivation methods and some of the principal advantages, assumptions, and limitations in the use of this sediment assessmenttool. Ideally, mean SQGQs should be included with other measures including results of toxicity tests and benthic community surveys to provide a weight of evidence when assessing the relative quality of contaminated sediments.
Assuntos
Monitoramento Ambiental/métodos , Sedimentos Geológicos/química , Poluentes Químicos da Água/análise , Poluição da Água/prevenção & controle , Abastecimento de Água , Animais , Guias como Assunto , Humanos , Medição de Risco , Testes de ToxicidadeRESUMO
A survey was designed and conducted to determine the severity, spatial patterns, and spatial extent of degraded sediment quality in Puget Sound (Washington State, USA). A weight of evidence compiled from results of chemical analyses, toxicity tests, and benthic infaunal analyses was used to classify the quality of sediments. Sediment samples were collected from 300 locations within a 2363 km(2) area extending from the US/Canada border to the inlets of southern Puget Sound and Hood Canal. Degraded conditions, as indicated with a combination of high chemical concentrations, significant toxicity, and adversely altered benthos, occurred in samples that represented about 1% of the total area. These conditions invariably occurred in samples collected within urbanized bays and industrial waterways, especially near the urban centers of Everett, Seattle, Tacoma, and Bremerton. Sediments with high quality (as indicated by no toxicity, no contamination, and the presence of a relatively abundant and diverse infauna) occurred in samples that represented a majority (68%) of the total study area. Sediments in which results of the three kinds of analyses were not in agreement were classified as intermediate in quality and represented about 31% of the total area. Relative to many other estuaries and marine bays of the USA, Puget Sound sediments ranked among those with minimal evidence of toxicant-induced degradation.
Assuntos
Sedimentos Geológicos/análise , Poluentes Químicos da Água/toxicidade , Aliivibrio fischeri/metabolismo , Anfípodes/efeitos dos fármacos , Animais , Linhagem Celular , Sistema Enzimático do Citocromo P-450/biossíntese , Monitoramento Ambiental , Indução Enzimática , Fertilização/efeitos dos fármacos , Genes Reporter/genética , Humanos , Luciferases/metabolismo , Luminescência , Metais/análise , Metais/toxicidade , Compostos Orgânicos/análise , Compostos Orgânicos/toxicidade , Ouriços-do-Mar/efeitos dos fármacos , Ouriços-do-Mar/fisiologia , Água do Mar , Silício/análise , Silício/toxicidade , Washington , Poluentes Químicos da Água/análiseRESUMO
Synoptic data on concentrations of sediment-associated chemical contaminants and benthic macroinfaunal community structure were collected from 1,389 stations in estuaries along the U.S. Atlantic and Gulf of Mexico coasts as part of the nationwide Environmental Monitoring and Assessment Program (EMAP). These data were used to develop an empirical framework for evaluating risks of benthic community-level effects within different ranges of sediment contamination from mixtures of multiple chemicals present at varying concentrations. Sediment contamination was expressed as the mean ratio of individual chemical concentrations relative to corresponding sediment quality guidelines (SQGs), including Effects Range-Median (ERM) and Probable Effects Level (PEL) values. Benthic condition was assessed using diagnostic, multi-metric indices developed for each of three EMAP provinces (Virginian, Carolinian, and Louisianian). Cumulative percentages of stations with a degraded benthic community were plotted against ascending values of the mean ERM and PEL quotients. Based on the observed relationships, mean SQG quotients were divided into four ranges corresponding to either a low, moderate, high, or very high incidence of degraded benthic condition. Results showed that condition of the ambient benthic community provides a reliable and sensitive indicator for evaluating the biological significance of sediment-associated stressors. Mean SQG quotients marking the beginning of the contaminant range associated with the highest incidence of benthic impacts (73-100% of samples, depending on the province and type of SQG) were well below those linked to high risks of sediment toxicity as determined by short-term toxicity tests with single species. Measures of the ambient benthic community reflect the sensitivities of multiple species and life stages to persistent exposures under actual field conditions. Similar results were obtained with preliminary data from the west coast (Puget Sound).