RESUMO
Based on the combination of ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF) and Waters UNIFI software, the chemical constituents of the classic prescription Xiaochengqi Decoction were qualitatively analyzed and identified. The UPLC conditions are as follows: Acquity HSS T3 reverse phase column(2.1 mm ×100 mm, 1.8 μm), column temperature of 30 ℃, mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B), and flow rate of 0.3 mL·min~(-1). High-resolution MS data of Xiaochengqi Decoction were collected in ESI~(+/-) modes by Fast DDA. The structures of the chemical constituents were tentatively characterized or identified by UNIFI software according to the retention time of reference standards and characteristic fragment ions in MS profile, and literature data. A total of 233 components in Xiaochengqi Decoction were identified, with 93 from wine-processed Rhei Radix et Rhizoma, 104 from bran-processed Aurantii Fructus Immaturus, and 36 from ginger-processed Magnoliae Officinalis Cortex. These 233 components included anthraquinones, flavonoids, lignans, alkaloids, coumarins, and phenylethanoid glycosides. The result provided experimental evidence for the further study on establishment of quality standard and product development of the formula.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , DDT/análogos & derivados , Medicamentos de Ervas Chinesas/química , Espectrometria de Massas , Rizoma/química , SoftwareRESUMO
Cationic liposome - CpG DNA complexes (lipoplexes) are known as stimulators of innate immunity via Toll-like receptor 9 (TLR9)-triggered activation of the nuclear factor kappa B (NF-κB) pathway. More recent reports suggest that DNA lipoplexes also engage DNA sensors in the cytosol leading to the stimulation of the interferon response factor (IRF) pathway. In this study a range of lipoplexes were formulated by using an invariable helper lipid, three different cationic lipids (DOTAP, DOTMA and DDA) and three different CpG-containing plasmids of different sizes. These lipoplexes exhibited similar hydrodynamic diameters, zeta-potentials and plasmid loading rates, despite the different lipid blends and CpG-containing plasmids. Binding and uptake of liposomal lipids by J774.A1 macrophages and JAWSII dendritic cells increased significantly (up to 4-fold) upon lipoplex formation. Cellular plasmid DNA uptake via lipoplexes compared to naked DNA was increased up to 18-fold. Analysis of signal transduction pathway activation in J774-DUAL™ reporter cells by liposomes or naked CpG plasmid DNA compared to their derived lipoplexes showed only minor activation of the NF-κB pathway, while the IRF pathway displayed massive activation factors of up to 46-fold. DOTAP- and DOTMA lipoplexes also led to massive interferon-alpha and -beta secretion of J774A.1 macrophages and JAWSII dendritic cells, which is a hallmark of IRF pathway activation. Cellular distribution studies on DOTAP lipoplexes suggest delivery of plasmid DNA via vesicular compartments into the cytosol. Taken together, the CpG plasmid DNA lipoplexes generated in this study appear to selectively stimulate DNA receptors activating the IRF pathway, while bypassing TLR9 and NF-κB activation.
Assuntos
DDT/análogos & derivados , DNA/metabolismo , Ácidos Graxos Monoinsaturados/metabolismo , Interferons/metabolismo , Plasmídeos/metabolismo , Compostos de Amônio Quaternário/metabolismo , Animais , Cátions , Linhagem Celular , DDT/administração & dosagem , DDT/metabolismo , DNA/administração & dosagem , Ácidos Graxos Monoinsaturados/administração & dosagem , Lipossomos , Camundongos , Plasmídeos/administração & dosagem , Compostos de Amônio Quaternário/administração & dosagem , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologiaRESUMO
The cationic dimethyldioctadecylammonium/trehalose 6,6,9-dibehenate (DDA/TDB) liposome is as a strong adjuvant system for vaccines, with remarkable immunostimulatory activity. The mucosal administration of vaccines is a potential strategy for inducing earlier and stronger mucosal immune responses to infectious diseases. In this study, we assessed whether the intranasal administration of cationic DDA/TDB liposomes combined with influenza antigen A (H3N2) can be used as a highly efficacious vaccine to induce mucosal and systemic antibody responses. Confocal laser scanning microscopy and a flow-cytometric analysis showed that the uptake of the cationic DDA/TDB liposome carrier was significantly higher than that of neutral 1,2-distearoyl-sn-glycero-3-phosphocholine/cholesterol (DSPC/Chol) or cationic 1,2-dioleoyl-3-trimethylammonium-propane/3ß-(N-[N',N'-dimethylaminoethane]-carbamoyl (DOTAP/DC-Chol) liposomes. Our results indicate that the cationic DDA/TDB liposome is more effective in facilitating its uptake by dendritic cells (DCs) in vitro than the DSPC/Chol or DOTAP/DC-Chol liposome. DCs treated with DDA/TDB liposomes strongly expressed CD80, CD86, and MHC II molecules, whereas those treated with DSPC/Chol or DOTAP/DC-Chol liposomes did not. C57BL/6 mice intranasally immunized with H3N2-encapsulating cationic DDA/TDB liposomes had significantly higher H3N2-specific s-IgA levels in their nasal wash fluid than those treated with other formulations. The DDA/TDB liposomes also simultaneously enhanced the serum IgG IgG2a, IgG1, and IgG2b antibody responses. In summary, DDA/TDB liposomes effectively facilitated their uptake by DCs and DCs maturation in vitro, and induced significantly higher mucosal IgA, systemic IgG, IgG1, and IgG2b antibody titres than other formulations after their intranasal administration in vivo. These results indicate that DDA/TDB liposomes are a promising antigen delivery carrier for clinical antiviral applications.
Assuntos
Adjuvantes Imunológicos/metabolismo , DDT/análogos & derivados , Células Dendríticas/metabolismo , Glicolipídeos/química , Imunidade Humoral/imunologia , Lipossomos/metabolismo , Mucosa/metabolismo , Animais , Anticorpos Antivirais/imunologia , Transporte Biológico , Fenômenos Químicos , DDT/química , Células Dendríticas/imunologia , Feminino , Vírus da Influenza A Subtipo H3N2/imunologia , Lipossomos/química , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Loss-of-function of barley mildew locus o (Mlo) confers durable broad-spectrum penetration resistance to the barley powdery mildew pathogen, Blumeria graminis f. sp. hordei (Bgh). Given the importance of mlo mutants in agriculture, surprisingly few molecular components have been identified to be required for this type of resistance in barley. With the aim to identify novel cellular factors contributing to mlo-based resistance, we devised a pharmacological inhibitor screen. Of the 41 rationally chosen compounds tested, five caused a partial suppression of mlo resistance in barley, indicated by increased levels of Bgh host cell entry. These chemicals comprise brefeldin A (BFA), 2',3'-dideoxyadenosine (DDA), 2-deoxy-d-glucose, spermidine, and 1-aminobenzotriazole. Further inhibitor analysis corroborated a key role for both anterograde and retrograde endomembrane trafficking in mlo resistance. In addition, all four ribonucleosides, some ribonucleoside derivatives, two of the five nucleobases (guanine and uracil), some guanine derivatives as well as various polyamines partially suppress mlo resistance in barley via yet unknown mechanisms. Most of the chemicals identified to be effective in partially relieving mlo resistance in barley also to some extent compromised powdery mildew resistance in an Arabidopsis mlo2 mlo6 double mutant. In summary, our study identified novel suppressors of mlo resistance that may serve as valuable probes to unravel further the molecular processes underlying this unusual type of disease resistance.
Assuntos
Agroquímicos/farmacologia , Resistência à Doença/efeitos dos fármacos , Resistência à Doença/genética , Hordeum/efeitos dos fármacos , Hordeum/genética , Proteínas de Plantas/genética , Agricultura/métodos , Brefeldina A/farmacologia , DDT/análogos & derivados , DDT/farmacologia , Desoxiglucose/farmacologia , Ribonucleosídeos/genética , Espermidina/farmacologia , Triazóis/farmacologiaRESUMO
The Palos Verdes Shelf (PVS) and the continental slope off the Palos Verdes Peninsula are highly contaminated by degradation products of the pesticide DDT (1-chloro-4-[2,2,2-trichloro-1-(4-chlorophenyl)ethyl]benzene). Sediment samples from two box cores were analyzed to obtain further information about the fate of DDT and its degradation products within the environment. After solvent extraction, an alkaline hydrolysis procedure was applied. A comprehensive screening for 26 DDT compounds revealed that DDT and its degradates contaminate not only the extractable fraction but also the fraction released by alkaline hydrolysis. A comparison of the quantitative distribution of DDT degradation products in the extractable fraction and released by alkaline hydrolysis showed a distinct difference. DDE (1-chloro-4-[2,2-dichloro-1-(4-chlorophenyl)ethenyl]benzene), DDD (1-chloro-4-[2,2-dichloro-1-(4-chlorophenyl)ethyl]benzene), DDMS (1-chloro-4-[2-chloro-1-(4-chlorophenyl)ethyl]benzene), and DDMU (1-chloro-4-[2-chloro-1-(4-chlorophenyl)ethenyl]benzene) were predominant in the sediment extracts but minor components of the hydrolyzable fraction. The most abundant compounds released by the alkaline hydrolysis were DBP (bis(4-chlorophenyl)methanone), DDNU (1-chloro-4-[1-(4-chlorophenyl)ethenyl]benzene), DDM (1-chloro-4-[(4-chlorophenyl)methyl]benzene) and the water-soluble DDA (2,2-bis(4-chlorophenyl)acetic acid). The release of DDA may point to the presence of an important degradation pathway in marine environments. Concentration levels of DDT-related compounds showed corresponding vertical profiles in both fractions, but were significantly lower in the fraction released by alkaline hydrolysis. In contrast to fluvial sediments contaminated by DDT and its degradates the alkaline hydrolysis products represented a minor portion of the total sedimentary burden in the analyzed marine sediments. These findings show the necessity of a comprehensive screening for all DDT isomers and breakdown products in the extractable and non-extractable fraction to assess the total contamination abundance and potential environmental risks.
Assuntos
DDT , Sedimentos Geológicos , California , DDT/análogos & derivados , DDT/análise , DDT/química , Diclorodifenil Dicloroetileno , Sedimentos Geológicos/análise , Sedimentos Geológicos/química , Hidrólise , SolubilidadeRESUMO
Abiotic and biotic processes can be used to remediate DDX (DDT, DDD, DDE, and DDNS) contaminated soils; these processes can be fostered using specific carbon-amendments to stimulate particular soil indigenous microbial communities to improve rates or extent of degradation. In this study, toluene and glycerol were evaluated as cosubstrates under aerobic and anoxic conditions to determine the degradation efficiencies of DDX and to elucidate possible degradation mechanisms. Slurry microcosms experiments were performed during 60 days using pretreated soil with zero-valent iron (ZVI). Toluene addition enhanced the percentage of degradation of DDX. DDNS was the main compound degraded (around 86%) under aerobic conditions, suggesting cometabolic degradation of DDX by toluene-degrading soil bacteria. Glycerol addition under anoxic conditions favored the abiotic degradation of DDX mediated by sulfate-reducing bacteria activity, where DDT was the main compound degraded (around 90%). The 16S rDNA metagenomic analyses revealed Rhodococcus ruber and Desulfosporosinus auripigmenti as the predominant bacterial species after 40 days of treatment with toluene and glycerol additions, respectively. This study provides evidence of biotic and abiotic DDX degradation by the addition of toluene and glycerol as cosubstrates in ZVI pretreated DDX-contaminated soil.
Assuntos
Biodegradação Ambiental , DDT/análogos & derivados , Glicerol/química , Ferro/química , Poluentes do Solo/análise , Tolueno/química , Aerobiose , Anaerobiose , DNA Bacteriano/genética , Microbiologia do SoloRESUMO
The fate of hydrophobic organic compounds in aquatic environment are largely determined by their exchange at sediment-water interface, which is highly dynamic and subject to rapidly evolving environmental conditions. In turn, environmental conditions may be governed by both physicochemical parameters and anthropogenic events. To examine the importance of various impact factors, passive sampling devices were deployed at the seafloor of Hailing Bay, an urbanized estuarine bay in Guangdong Province of South China to measure the sediment-water diffusion fluxes of several metabolites of dichlorodiphenyltrichloroethane (DDT), p,p'-DDE, p,p'-DDD and o,p'-DDD. The physicochemical properties of water (temperature, pH, salinity and dissolved oxygen) and surface sediment (sediment organic matter, physical composition, pH, water content, colony forming unit and catalase activity) were also measured. The results showed that the diffusion fluxes of o,p'-DDD, p,p'-DDD and p,p'-DDE at sites A1 and A2 near a fishing boat maintenance facility ranged from 0.42 to 4.73 ng m-2 d-1 (from sediment to overlying water), whereas those at offshore sites varied between -0.03 and -3.02 ng m-2 d-1 (from overlying water to sediment), implicating A1 and A2 as the sources of the target compounds. The distribution patterns of the diffusion fluxes of the target compounds were different from those of water and sediment parameters (water temperature, salinity, sediment texture, pH, colony forming unit and catalase activity) at six sampling sites. This finding suggested that none of these parameters were critical in dictating the sediment-water diffusion fluxes. Besides, decreases in the contents of kerogen and black carbon by 6.7% and 11% would enhance the diffusion fluxes of the target compounds by 11-14% and 12-23%, respectively, at site A1, indicating that kerogen and black carbon were the key factors in mediating the sediment-water diffusion fluxes of DDT-related compounds in field environments.
Assuntos
Baías/química , DDT/análogos & derivados , DDT/análise , Água Doce/química , Sedimentos Geológicos/química , Urbanização , Poluentes Químicos da Água/análise , China , Diclorodifenildicloroetano/análise , Difusão , Monitoramento Ambiental , Hidrocarbonetos Clorados/análiseRESUMO
The temporal-spatial distributions of DDT-related contaminants (DDXs), including DDT (dichlorodiphenyltrichloroethane), DDE (dichlorodiphenyldichloroethylene) and DDD (dichlorodiphenyldichloroethane), in the sediments of Lake Chaohu and their influencing factors were studied. p,p-DDE and p,p-DDD were found to be the two dominant components of DDXs in both surface and core sediments. The parent DDT compounds were still detectable in sediment cores after the late 1930s. Historical usage of technical DDT was identified as the primary source of DDXs in sediments, as indicated by DDT/(DDD + DDE) ratios of less than one. The residual levels of DDXs were higher in the surface and core sediments in the western lake area than in other lake areas, which might be due to the combined inflow effects of municipal sewage, industrial wastewater and agricultural runoff. The DDX residues in the sediment cores reached peak values in the late 1970s or early 1980s. There were significant positive relationships between DDX residues in sediment cores with annual DDT production and with fine particulate sizes (<4.5 µm). The relationship between the DDXs and TOC in sediment was complex, as indicated by the significant differences among the surface and core sediments. The algae-derived organic matter significantly influenced the amount of residue, composition and distribution of DDXs in the sediments. The DDD/DDE ratios responded well to the anaerobic conditions in the sediments that were caused by algal blooms after the late 1970s in the western lake area. This suggests that the algae-derived organic matter was an important factor and served as a biomarker of eutrophication and also affected the DDX residues and lifecycle in the lake ecosystem.
Assuntos
DDT/análogos & derivados , DDT/análise , Sedimentos Geológicos/química , Hidrocarbonetos Clorados/análise , Lagos/química , Poluentes Químicos da Água/análise , China , Monitoramento Ambiental , EutrofizaçãoRESUMO
The incidence of autoimmune Type 1 diabetes (T1D) has been steadily rising in developed countries. Although the exact cause of T1D remains elusive, it is known that both genetics and environmental factors play a role in its immunopathogenesis. Whereas a positive association between p,p'-DDE, a DDT metabolite, and Type 2 diabetes (T2D) has been well established, its role in T1D development in an experimental animal model has never been elucidated. This study seeks to investigate the effects of DDE exposure on the development of T1D in a NOD mouse model. As T1D is a T-cell-mediated disease, the underlying mechanism of DDE action on T-cells was studied in vitro and, in the context of acute and chronic DDE exposure, in vivo by investigating lymphocytes' viability, proliferation, their subsets and cytokine profiles. Chronic high-dose DDE treatment, initiated in pre-diabetic 8-week-old NOD females administered twice weekly intraperitoneally with 50 mg/kg DDE, significantly increased diabetes incidence and augmented disease severity in treated animals. Whereas T-cell proliferation and cell viability in the spleens of treated mice were not affected, diabetogenic action of chronic DDE exposure was associated with a decrease in regulatory T-cells and a suppression of secretion of protective cytokines, such as IL-4 and IL-10. Interestingly, an acute high-dose in vivo treatment of 8-week-old NOD males with 100 mg DDE/kg, administered intraperitoneally every other day over a period of 10 days, increased T-cell proliferation and potentiated pro-inflammatory and TH1-type cytokine secretion, without affecting the splenocytes viability and the T-cell sub-populations. These results confirm that high-dose DDE treatments affect the immune system, in particularly T-cell function. In conclusion, this study shows for the first time that high-dose chronic DDE exposure exhibits a diabetogenic potential, with an underlying immunomodulatory mechanism of action, in the development of T1D in an experimental mouse NOD model.
Assuntos
Doenças Autoimunes/imunologia , DDT/toxicidade , Diabetes Mellitus Tipo 1/imunologia , Diclorodifenil Dicloroetileno/toxicidade , Poluentes Ambientais/toxicidade , Subpopulações de Linfócitos T/efeitos dos fármacos , Linfócitos T Reguladores/efeitos dos fármacos , Animais , Células Cultivadas , Citocinas/metabolismo , DDT/análogos & derivados , Modelos Animais de Doenças , Progressão da Doença , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos NOD , Subpopulações de Linfócitos T/imunologia , Linfócitos T Reguladores/imunologia , Equilíbrio Th1-Th2/efeitos dos fármacosRESUMO
Human exposure to persistent organic pollutants (POPs) is a certainty, even to long banned pesticides like o,p'-dichlorodiphenyltrichloroethane (o,p'-DDT), and its metabolites p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE), and p,p'-dichlorodiphenyldichloroethane (p,p'-DDD). POPs are known to be particularly toxic and have been associated with endocrine-disrupting effects in several mammals, including humans even at very low doses. As environmental estrogens, they could play a critical role in carcinogenesis, such as in breast cancer. With the purpose of evaluating their effect on breast cancer biology, o,p'-DDT, p,p'-DDE, and p,p'-DDD (50-1000 nM) were tested on two human breast adenocarcinoma cell lines: MCF-7 expressing estrogen receptor (ER) α and MDA-MB-231 negative for ERα, regarding cell proliferation and viability in addition to their invasive potential. Cell proliferation and viability were not equally affected by these compounds. In MCF-7 cells, the compounds were able to decrease cell proliferation and viability. On the other hand, no evident response was observed in treated MDA-MB-231 cells. Concerning the invasive potential, the less invasive cell line, MCF-7, had its invasion potential significantly induced, while the more invasive cell line MDA-MB-231, had its invasion potential dramatically reduced in the presence of the tested compounds. Altogether, the results showed that these compounds were able to modulate several cancer-related processes, namely in breast cancer cell lines, and underline the relevance of POP exposure to the risk of cancer development and progression, unraveling distinct pathways of action of these compounds on tumor cell biology.
Assuntos
Neoplasias da Mama/induzido quimicamente , Neoplasias da Mama/patologia , Poluentes Ambientais/toxicidade , Hidrocarbonetos Clorados/toxicidade , Praguicidas/toxicidade , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , DDT/análogos & derivados , DDT/toxicidade , DNA/biossíntese , DNA/genética , Disruptores Endócrinos/toxicidade , Receptor alfa de Estrogênio/efeitos dos fármacos , Feminino , Humanos , Células MCF-7 , Invasividade Neoplásica/patologia , Timidina/análogos & derivados , Timidina/metabolismoRESUMO
DDT has enjoyed the reputation of a successful pesticide in disease control programme and agricultural practices along with the serious opposition and ban later on due to its biomagnification and toxic action against non-target organisms. The present work was carried out to develop QSAR models for analysing DDT analogues for their pesticidal activity and in silico validation of these models. A 2D-QSAR model was generated using stepwise with multiple regression, and the model with a value of r(2) = 0.7324; q(2) = 0.6215; pred r(2) = 0.7038, containing five descriptors, was selected for further study. The 3D QSAR with CoMFA analysis showed that steric contribution of 21% and electrostatic contribution of about 79% were required for larvicidal activity of DDT analogues. A set of 3430 molecules was generated using the basic DDT skeleton as template, and these were evaluated for their mosquito larvicidal activity using the generated QSAR models and DDT as standard. Eleven molecules were selected for in silico validation of these models. For this, a docking study of the selected molecules against the homology-modelled voltage-gated sodium channel of Anopheles funestus was conducted. The study showed that the activities of these analogues as predicted by 2D-QSAR, 3D-QSAR with CoMFA and dock score were observed to be well correlated.
Assuntos
Anopheles/efeitos dos fármacos , DDT/química , Inseticidas/química , Relação Quantitativa Estrutura-Atividade , Canais de Sódio/metabolismo , Animais , Simulação por Computador , DDT/análogos & derivados , DDT/farmacologia , Inseticidas/farmacologia , Larva/efeitos dos fármacos , Modelos Moleculares , Eletricidade EstáticaRESUMO
The purpose of this study was to develop a rapid and cost efficient hair extraction method, using the headspace solid phase microextraction (HSSPME) technique for the simultaneous determination and biomonitoring of 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane) (DDT) and its isomers/metabolites and polychlorinated biphenyls (PCBs) in hair samples. A total of 72 head hair samples were collected from children living in urban and rural regions of the island of Crete. Two hundred milligrams of hair were digested under alkaline conditions and thermostated for 30 min at 90°C while a 65 µm PDMS/DVB fibre was exposed into the headspace of the vial. Analytical parameters of the method (time of incubation, agitation speed, recovery, precision, accuracy, carry over, matrix effect, linearity, and selectivity) were examined. Recoveries of the DDTs in the spiked hair samples were calculated from 42.3% for opDDD to 87.1% for opDDE, while recoveries for PCB congeners were from 52.6% for PCB138 to 96.6 % for PCB28. The method was applied for the analysis of authentic hair samples. Significant differences (p=0.001) of the burden to total DDTs (sumDDTs) as well as of the frequencies of detection of positive samples (p=0.020) were observed between the examined regions. Moreover, significant differences in the detected concentrations of PCB congeners were observed for PCB52 (p<0.001) and PCB28 (p=0.017) as well for their prevalence between urban and rural regions. Application of HSSPME for the biomonitoring of DDTs and PCBs biomarkers in hair was tested and successfully applied to the analysis of spiked and authentic hair samples. HSSPME was found to be substantially simpler and faster procedure than previous reported sample treatment procedures.
Assuntos
DDT/análise , Monitoramento Ambiental/métodos , Cabelo/química , Bifenilos Policlorados/análise , Microextração em Fase Sólida/métodos , Criança , Pré-Escolar , DDT/análogos & derivados , Exposição Ambiental/análise , Feminino , Cromatografia Gasosa-Espectrometria de Massas/métodos , Grécia , Humanos , Masculino , População Rural , Fatores de Tempo , População UrbanaRESUMO
Recent understanding of the pathogenesis of tuberculosis allows the possible application of immunotherapy for the treatment of tuberculosis. Therapies that would upregulate the host antimycobacterial immune response and/or attenuate T-cell suppressive and macrophage-deactivating cytokines may prove to be useful in the treatment of tuberculosis. ESAT6, 6-kDa early secreted antigenic target, is a potent protective antigen and is considered as major target for long-lived memory cells. In the present study the immunotherapeutic potential of ESAT-6 has been evaluated in mouse model of experimental tuberculosis. In the present study the ESAT-6 protein was cloned in Escherichia coli using pET23a(+) plasmid and purified by Ni(2+)-NTA chromatography. Further, the immunotherapeutic potential of the recombinant ESAT-6 (in terms of CFU enumeration in the target organs and histopathological analysis of lungs) was evaluated against experimental tuberculosis. The recombinant ESAT-6 with C-terminal histidine-tag and free N-terminus mimics the natural form of ESAT-6 has been successfully cloned and purified. The recombinant ESAT-6 protein adjuvanted with dimethyl dioctadecylammonium bromide (DDA) moderately reduced the bacterial load in the target organs of infected mice. Further, the formulation (ESAT-6-DDA) was able to act synergistically when given in combination with antituberculosis drugs. This recombinant ESAT-6 showed good immunotherapeutic potential against experimental tuberculosis and can be used as an adjunct to the conventional antituberculosis chemotherapy.
Assuntos
Antígenos de Bactérias/administração & dosagem , Proteínas de Bactérias/administração & dosagem , Escherichia coli/genética , Imunoterapia/métodos , Pulmão/imunologia , Mycobacterium tuberculosis/imunologia , Proteínas Recombinantes/administração & dosagem , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/terapia , Animais , Antígenos de Bactérias/imunologia , Antituberculosos/uso terapêutico , Carga Bacteriana/efeitos dos fármacos , Proteínas de Bactérias/imunologia , Clonagem Molecular , Ensaio de Unidades Formadoras de Colônias , DDT/administração & dosagem , DDT/análogos & derivados , Modelos Animais de Doenças , Sinergismo Farmacológico , Feminino , Humanos , Pulmão/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/imunologiaRESUMO
The presence of endocrine disruptors in source water is of great concern because of their suspected adverse effects on humans, even when present at very low levels. As the main source of potable water supply, rivers in Malaysia are highly susceptible to contamination by various endocrine disruptors originating from anthropogenic activities. In this study, the contamination levels of 1,1,1-trichloro-2,2-bis (4-chlorophenyl) ethane (DDT) and its metabolites and di-(2-ethylhexyl) phthalate (DEHP) in rivers of Selangor were examined using gas chromatography-mass spectrometry. Samples were collected from sites representing source water for 18 drinking water treatment plants in Selangor between July 2008 and July 2009. DDT and its metabolites were detected in only 14% of the 192 samples analysed at levels ranging from 0.6 to 14.6 ng/L. Meanwhile DEHP was detected in 96.8% of the samples at levels ranging from below quantitation level (18 ng/L) to 970 ng/L. The detected levels of DDTs and DEHP were lower than the WHO and Malaysian Guidelines for Drinking Water Quality. Data obtained from this study should also serve as a reference point for future surveillance on these endocrine disruptors.
Assuntos
DDT/análogos & derivados , Dietilexilftalato/química , Disruptores Endócrinos/química , Poluentes Químicos da Água/química , DDT/química , Cromatografia Gasosa-Espectrometria de Massas , Malásia , Rios/químicaRESUMO
Organochlorine pesticides (OCPs) are persistent endocrine disruptors. OCPs cross the placenta; this prenatal exposure has been associated with adverse pregnancy outcomes. We investigated associations between prenatal exposure to OCPs and gestational age and birth weight in 600 infants born between 1960 and 1963. The primary OCP was 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (p,p'-DDT), its primary metabolite, 1,1'-dichloro-2,2'-bis(p-chlorophenyl)ethylene (p,p'-DDE) and the contaminant, 1,1,1-trichloro-2-(p-chlorophenyl)-2-(o-chlorophenyl)-ethane (o,p'-DDT). Regression analysis indicated that for each natural log unit increase in p,p'-DDT, birth weight increased by 274 g (95% CI: 122, 425) when controlling for p,p'-DDE and o,p'-DDT. At a given level of p,p'-DDT exposure, o,p'-DDT and p,p'-DDE were associated with decreased birth weight. p,p'-DDE was negatively associated with length of gestation, controlling for p,p'-DDT and o,p'-DDT. These findings suggest opposing associations between exposure to p,p'-DDT and p,p'-DDE and birth weight. We did not find evidence to support mediation by maternal thyroid hormone status nor that the association differed by sex.
Assuntos
Peso ao Nascer , DDT/sangue , Diclorodifenil Dicloroetileno/sangue , Disruptores Endócrinos/sangue , Exposição Materna/efeitos adversos , Adolescente , Adulto , DDT/análogos & derivados , Feminino , Idade Gestacional , Humanos , Recém-Nascido , Masculino , Gravidez/sangue , Adulto JovemRESUMO
The purpose of this study was to investigate the effect of 1,1,1-trichloro-2,2-bis-(chlorophenyl)ethane (DDT), 1,1-bis-(chlorophenyl)-2,2-dichloroethene (DDE), and 1,1-dichloro-2,2-bis(chlorophenyl)ethane (DDD) isomers on COX-2 expression in a human trophoblast-derived cell line. Cultured HTR-8/SVneo trophoblast cells were exposed to DDT isomers and its metabolites for 24 h, and COX-2 mRNA and protein expression were assessed by RT-PCR, Western blotting, and ELISA. Prostaglandin E2 production was also measured by ELISA. Both COX-2 mRNA and protein were detected under control (unexposed) conditions in the HTR-8/SVneo cell line. COX-2 protein expression and prostaglandin E2 production but not COX-2 mRNA levels increased only after DDE and DDD isomers exposure. It is concluded that DDE and DDD exposure induce the expression of COX-2 protein, leading to increased prostaglandin E2 production. Interestingly, the regulation of COX-2 by these organochlorines pesticides appears to be at the translational level.
Assuntos
Carcinógenos Ambientais/toxicidade , Ciclo-Oxigenase 2/biossíntese , DDT/toxicidade , Diclorodifenil Dicloroetileno/toxicidade , Diclorodifenildicloroetano/toxicidade , Inseticidas/toxicidade , Trofoblastos/efeitos dos fármacos , Carcinógenos Ambientais/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , DDT/análogos & derivados , DDT/metabolismo , Diclorodifenil Dicloroetileno/análogos & derivados , Diclorodifenil Dicloroetileno/metabolismo , Diclorodifenildicloroetano/análogos & derivados , Diclorodifenildicloroetano/metabolismo , Dinoprostona/metabolismo , Indução Enzimática/efeitos dos fármacos , Feminino , Humanos , Inseticidas/química , Inseticidas/metabolismo , Concentração Osmolar , Biossíntese de Proteínas/efeitos dos fármacos , RNA Mensageiro/metabolismo , Estereoisomerismo , Trofoblastos/enzimologia , Trofoblastos/metabolismoRESUMO
Dichlorodiphenyltrichloroethane (DDT) is ubiquitous in the environment, and the exposure to DDT and its related pesticides has long been linked to endocrine disruption. The mechanism of endocrine disruption toward targeted receptors, however, remains unclear. Probing the molecular recognition of DDT analogs by targeted receptors at the atomic level is critical for deciphering this mechanism. Molecular dynamics (MD) simulations were applied to probe the molecular recognition process of DDT and its five analogs, including dichlordiphenyldichloroethylene (DDE), dichlorodiphenyldichloroethane (DDD), methoxychlor (MXC), p,p'-hydroxy-DDT (HPTE), and dicofol by human estrogen receptor (ER) α and human ER-related receptor (ERR) γ. Van der Waals interactions mainly drive the interactions of DDT analogs with ERα ligand-binding domain (LBD) and ERRγ LBD. Minor structural changes of DDT analogs in the number and position of chlorine and phenolic hydroxyl moiety cause differences in binding modes through aromatic stacking and hydrogen bonding and thus affect differently conformational changes of ERα LBD and ERRγ LBD. The binding of DDT analogs affects the helix 12 orientation of ERα LBD but causes no rearrangement of helix 12 of ERRγ LBD. These results extend our understanding of how DDT analogs exert their estrogen-disrupting effects toward different receptors via multiple mechanisms.
Assuntos
DDT/toxicidade , Disruptores Endócrinos/toxicidade , Receptor alfa de Estrogênio/efeitos dos fármacos , Receptores de Estrogênio/efeitos dos fármacos , DDT/análogos & derivados , Diclorodifenil Dicloroetileno/toxicidade , Diclorodifenildicloroetano/toxicidade , Humanos , Ligação de Hidrogênio , Metoxicloro/toxicidade , Conformação Molecular , Simulação de Dinâmica Molecular , Praguicidas , Fenóis/toxicidade , Ligação Proteica , Estrutura Secundária de ProteínaRESUMO
DDT, DDE and some additional lipophilic derivatives are recognized contaminants in sediments of Southern California Bight (SCB). Only about 10% of total DDTs discharged into the SCB are accounted for using available monitoring data (sediment, water, and biota). DDA represented up to 0.03% DDTs (DDT/DDE/DDD) in SCB surface sediments (top 2 cm) in amounts up to 76 µg DDA/kg dry weight. Highest DDA levels were found where DDT and DDD levels were maximal at the primary wastewater outfall indicating a natural precursor-product relationship for DDT and DDA. Still culture of SCB sediment revealed limited DDA formation following DDT fortification. DDA residues have also been found in contaminated Long Island, NY sediments provided by USGS. The formation of DDA and its potential release from sediments may be significant in resolution of uncertainties concerning the natural recovery of sediments in DDT-contaminated environments.
Assuntos
DDT/análogos & derivados , DDT/análise , Sedimentos Geológicos/química , Inseticidas/análise , Poluentes Químicos da Água/análise , California , Monitoramento AmbientalRESUMO
The organochlorine insecticide DDT (1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane) is still used for malaria vector control in certain areas of South Africa. The strict Stockholm Convention on Persistent Organic Pollutants (POPs) allows spraying on the inside of traditional dwellings with DDT. In rural villages contaminated dust presents an additional pathway for exposure to DDT. We present a new method for the determination of DDT in indoor air where separate vapour and particulate samples are collected in a single step with a denuder configuration of a multi-channel open tubular silicone rubber (polydimethylsiloxane (PDMS)) trap combined with a micro quartz fibre filter. The multi-channel PDMS trap section of the denuder concentrates vapour phase insecticide whereas particle associated insecticide is transferred downstream where it is collected on a micro-fibre filter followed by a second multi-channel PDMS trap to capture the blow-off from the filter. The multi-channel PDMS trap and filter combination are designed to fit a commercial thermal desorber for direct introduction of samples into a GC-MS. The technique is solvent-free. Analyte extraction and sample clean-up is not required. Two fractions, vapour phase and particulate phase p,p'-DDT, o,p'-DDT; p,p'-DDD, o,p'-DDD; p,p'-DDE and o,p'-DDE in 4 L contaminated indoor air, were each quantitatively analysed by GC-MS using isotopically labelled ring substituted (13)C(12) -p,p'-DDT as an internal standard. Limits of detection were 0.07-0.35 ng m(-3) for p,p'-DDT, o,p'-DDT, p,p'-DDD, o,p'-DDD, p,p'-DDE and o,p'-DDE. Ratios of airborne p,p'-DDD/p,p'-DDT and of o,p'-DDT/p,p'-DDT are unusual and do not match the ideal certified ingredient composition required of commercial DDT. Results suggest that the DDT products used for indoor residual spraying (IRS) prior to, and during 2007, may have been compromised with regards to insecticidal efficacy, demonstrating the power of this new environmental forensics tool.
Assuntos
Poluição do Ar em Ambientes Fechados/análise , DDT/análise , Inseticidas/análise , Malária/prevenção & controle , Absorção , DDT/análogos & derivados , Diclorodifenil Dicloroetileno/análise , Diclorodifenildicloroetano/análise , Dimetilpolisiloxanos/química , Poeira/análise , Monitoramento Ambiental , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Limite de Detecção , África do Sul , VolatilizaçãoRESUMO
BACKGROUND, AIM, AND SCOPE: 2,2-bis(chlorophenyl)-1,1,1-trichloroethane (DDT) metabolites, other than those routinely measured [i.e., 2,2-bis(chlorophenyl)-1,1-dichloroethylene (DDE) and 2,2-bis(chlorophenyl)-1,1-dichloroethane (DDD)], have recently been detected in elevated concentrations not only in the surface water of Teltow Canal, Berlin, but also in sediment samples from Elbe tributaries (e.g., Mulde and Havel/Spree). This was paralleled by recent reports that multiple other metabolites could emerge from the degradation of parent DDT by naturally occurring organisms or by interaction with some heavy metals. Nevertheless, only very few data on the biological activities of these metabolites are available to date. The objective of this communication is to evaluate, for the first time, the cytotoxicity, dioxin-like activity, and estrogenicity of the least-studied DDT metabolites. METHODS: Four DDT metabolites, p,p'-2,2-bis(chlorophenyl)-1-chloroethylene (DDMU), p,p'-2,2-bis(chlorophenyl)-1-chloroethane (DDMS), p,p'-2,2-bis(4-ch1oropheny1)acetonitrile (DDCN), and p,p'-2,2-bis(chlorophenyl)acetic acid (DDA), were selected based on their presence in environmental samples in Germany such as in sediments from the Mulde River and Teltow Canal. O,p'-DDT was used as reference in all assays. Cytotoxicity was measured by neutral red retention with the permanent cell line RTG-2 of rainbow trout (Oncorhynchus mykiss). Dioxin-like activity was determined using the 7-ethoxyresorufin-O-deetylase assay. The estrogenic potential was tested in a dot blot/RNAse protection-assay with primary hepatocytes from male rainbow trout (O. mykiss) and in a yeast estrogen screen (YES) assay. RESULTS: All DDT metabolites tested revealed a clear dose-response relationship for cytotoxicity in RTG-2 cells, but no dioxin-like activities with RTL-W1 cells. The dot blot/RNAse protection-assay demonstrated that the highest non-toxic concentrations of these DDT metabolites (50 µM) had vitellogenin-induction potentials comparable to the positive control (1 nM 17ß-estradiol). The estrogenic activities could be ranked as o,p'-DDT > p,p'-DDMS > p,p'-DDMU > p,p'-DDCN. In contrast, p,p'-DDA showed a moderate anti-estrogenic effect. In the YES assay, besides the reference o,p'-DDT, p,p'-DDMS and p,p'-DDMU displayed dose-dependent estrogenic potentials, whereas p,p'-DDCN and p,p'-DDA did not show any estrogenic potential. DISCUSSION: The reference toxicant o,p'-DDT displayed a similar spectrum of estrogenic activities similar to 17ß-estradiol, however, with a lower potency. Both p,p'-DDMS and p,p'-DDMU were also shown to have dose-dependent estrogenic potentials, which were much lower than the reference o,p'-DDT, in both the vitellogenin and YES bioassays. Interestingly, p,p'-DDA did not show estrogenic activity but rather displayed a tendency towards anti-estrogenic activity by inhibiting the estrogenic effect of 17ß-estradiol. The results also showed that the p,p'-metabolites DDMU, DDMS, DDCN, and DDA do not show any dioxin-like activities in RTL-W1 cells, thus resembling the major DDT metabolites DDD and DDE. CONCLUSIONS: All the DDT metabolites tested did not exhibit dioxin-like activities in RTL-W1 cells, but show cytotoxic and estrogenic activities. Based on the results of the in vitro assays used in our study and on the reported concentrations of DDT metabolites in contaminated sediments, such substances could, in the future, pose interference with the normal reproductive and endocrine functions in various organisms exposed to these chemicals. Consequently, there is an urgent need to examine more comprehensively the risk of environmental concentrations of the investigated DDT metabolites using in vivo studies. However, this should be paralleled also by periodic evaluation and monitoring of the current levels of the DDT metabolites in environmental matrices. RECOMMENDATIONS AND PERSPECTIVES: Our results clearly point out the need to integrate the potential ecotoxicological risks associated with the "neglected" p,p'-DDT metabolites. For instance, these DDT metabolites should be integrated into sediment risk assessment initiatives in contaminated areas. One major challenge would be the identification of baseline data for such risk assessment. Further studies are also warranted to determine possible additive, synergistic, or antagonistic effects that may interfere with the fundamental cytotoxicity and endocrine activities of these metabolites. For a more conclusive assessment of the spectrum of DDT metabolites, additional bioassays are needed to identify potential anti-estrogenic, androgenic, and/or anti-androgenic effects.