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In continuation to our studies on radioresistance in meningioma, here we show that radiation treatment (7 Gy) induces G2/M cell cycle arrest in meningioma cells. Phosphorylation of Chk2, Cdc25c and Cdc2 were found to be key events since interference with Chk2 activation and cyclin B1/Cdc2 interaction led to permanent arrest followed by apoptosis. Irradiated cells showed recovery and formed aggressive intracranial tumors with rapid spread and morbidity. Nevertheless, knock down of uPAR with or without radiation induced permanent arrest in G2/M phase and subsequent apoptosis in vitro and in vivo . In conclusion, our data suggest that combination treatment with radiation and uPAR knock down or other inhibitors resulting in non-reversible G2/M arrest may be beneficial in the management of meningiomas.

We present results from the OP3 campaign in Sabah during 2008 that allow us to study the impact of local emission changes over Borneo on atmospheric composition at the regional and wider scale. OP3 constituent data provide an important constraint on model performance. Treatment of boundary layer processes is highlighted as an important area of model uncertainty. Model studies of land-use change confirm earlier work, indicating that further changes to intensive oil palm agriculture in South East Asia, and the tropics in general, could have important impacts on air quality, with the biggest factor being the concomitant changes in NO x emissions. With the model scenarios used here, local increases in ozone of around 50 per cent could occur. We also report measurements of short-lived brominated compounds around Sabah suggesting that oceanic (and, especially, coastal) emission sources dominate locally. The concentration of bromine in short-lived halocarbons measured at the surface during OP3 amounted to about 7 ppt, setting an upper limit on the amount of these species that can reach the lower stratosphere.

Estrogen regulates numerous developmental and physiological processes. Most effects are mediated by estrogen receptors (ERs), which function as ligand-regulated transcription factors. Estrogen also regulates the activity of GPR30, a membrane-associated G protein-coupled receptor. Many different types of environmental contaminants can activate ERs; some can bind GPR30 as well. There is growing concern that exposure to some of these compounds, termed xenoestrogens, is interfering with the behavior and reproductive potential of numerous wildlife species, as well as affecting human health. Here, we investigated how two common, environmentally pervasive chemicals affect the in vivo expression of a known estrogen target gene in the brain of developing zebrafish embryos, aromatase AroB, which converts androgens to estrogens. We confirm that, like estrogen, the well-studied xenoestrogen bisphenol A (BPA, a plastics monomer), induces strong brain-specific overexpression of aromatase. Experiments using ER- and GPR30-selective modulators argue that this induction is largely through nuclear ERs. BPA induces dramatic overexpression of AroB RNA in the same subregions of the developing brain as estrogen. The antibacterial triclocarban (TCC) by itself stimulates AroB expression only slightly, but TCC strongly enhances the overexpression of AroB that is induced by exogenous estrogen. Thus, both BPA and TCC have the potential to elevate levels of aromatase and, thereby, levels of endogenous estrogens in the developing brain. In contrast to estrogen, BPA-induced AroB overexpression was suppressed by TCC. These results indicate that exposures to combinations of certain hormonally active pollutants can have outcomes that are not easily predicted from their individual effects.

The antimicrobial triclocarban (TCC) is frequently found in personal care products and commonly observed in surface waters and sediments. Due to its long environmental persistence TCC accumulates in sewage sludge. It also shows a high unintended biological activity as a potent inhibitor of the soluble epoxide hydrolase (sEH) and may be an endocrine disruptor. In this study, we investigated bioconcentration, metabolism and elimination of TCC in fish using Medaka (Oryzias latipes) as a model. Medaka larvae (7±1 days post hatching) were exposed to 63 nM (20 µg/L) TCC water for 24 hours. The LC-MS/MS analysis of water and tissues provided bioconcentration of TCC and its metabolites in fish body and rapid excretion into culture water. Results from tissue samples showed a tissue concentration of 34 µmol/kg and a log bioconcentration factor (BCF) of 2.86. These results are slightly lower than previous findings in snails and algae. A significant portion of the absorbed TCC was oxidatively metabolized by the fish to hydroxylated products. These metabolites underwent extensive phase II metabolism to yield sulfate and glucuronic acid conjugates. The most abundant metabolite in fish tissue was the glucuronide of 2’-OH-TCC. Elimination of TCC after transferring the fish to fresh water was rapid, with a half-life of 1 hour. This study shows that larval medaka metabolize TCC similarly to mammals. The rapid rate of metabolism results in a lower bioconcentration than calculated from the n-octanol/water partition coefficient of TCC.

article The increasing use of the antimicrobial triclocarban (TCC) in personal care products (PCPs) has resulted in concern regarding environmental pollution. TCC is a potent inhibitor of soluble epoxide hydrolase (sEH). Inhibitors of sEH (sEHIs) are anti-inflammatory, anti-hypertensive and cardio-protective in multiple animal models. However, the in vivo effects anticipated from a sEHI have not been reported for TCC. Here we demonstrated the anti-inflammatory effects in vivo of TCC in a murine model. TCC was employed in a lipopolysaccharide (LPS)-challenged murine model. Systolic blood pressure, plasma levels of several inflammatory cytokines and chemokine, and metabolomic profile of plasma oxylipins were determined. TCC significantly reversed LPS-induced morbid hypotension in a time-dependent manner. TCC significantly repressed the increased release of inflammatory cytokines and chemokine caused by LPS. Furthermore, TCC significantly shifted the oxylipin profile in vivo in a time-dependent manner towards resolution of inflammation as expected from a sEHI. These results demonstrated that at the doses used TCC is anti- inflammatory in the murine model. This study suggests that TCC may provide some benefits in humans in addition to its antimicrobial activities due to its potent inhibition of sEH. It may be a promising starting point for developing new low volume high value applications of TCC. However these biological effects also caution against the general over use of TCC in PCPs.

This study investigated the occurrence of 43 emerging contaminants including 9 endocrine-disrupting chemicals and 34 pharmaceuticals in three sites in Hebei Province, north China. Each site has a wastewater irrigated plot and a separate groundwater irrigated plot for comparison purpose. The results showed that the concentrations of the target compounds in the wastewater irrigated soils were in most cases higher than those in the groundwater irrigated soils. Among the 43 target compounds, nine compounds bisphenol-A, triclocarban, triclosan, 4-nonylphenol, salicylic acid, oxytetracycline, tetracycline, trimethoprim and primidone were detected at least once in the soils. Preliminary environmental risk assessment showed that triclocarban might pose high risks to terrestrial organisms while the other detected compounds posed minimal risks. Irrigation with wastewater could lead to presence or accumulation of some emerging contaminants to some extent in irrigated soils.

Triclosan (TCS) and triclocarban (TCC) are widely used broad spectrum bactericides that are common pollutants of waterways and soils. Methyl triclosan (mTCS) is the predominant bacterial TCS metabolite. Previous studies have shown that TCS disrupts thyroid hormone (TH) action; however, the effects of mTCS or TCC are not known. The present study uses the cultured frog tadpole tail fin biopsy (C-fin) assay and the TH-responsive rat pituitary GH3 cell line to assess the effects of these three chemicals (1-1000 nM) on TH signaling and cellular stress within 48 h. mRNA abundance of TH receptor β, Rana larval keratin type I (TH-response), heat shock protein 30, and catalase (stress-response) was measured using quantitative real-time polymerase chain reaction in the C-fin assay. The TH-responsive gene transcripts encoding growth hormone, deiodinase I, and prolactin were measured in GH3 cells with the heat shock protein 70 transcript acting as a cellular stress indicator. We found alteration of stress indicators at a wide range of concentrations of TCS, mTCS, and TCC in both test systems. mTCS and TCC affected TH-responsive gene transcripts at the highest concentration in mammalian cells, whereas a modest effect included lower concentrations in the C-fin assay. In contrast, TCS did not affect TH-responsive transcripts. These results identify nontarget biological effects of these bacteriocides on amphibian and mammalian cells and suggest the TH-disrupting effects observed for TCS could be mediated through its metabolite.

Triclocarban (TCC; 3,4,4′-trichlorocarbanilide) is an antimicrobial agent used widely in various personal hygiene products including soaps. Recently, TCC has been shown to enhance testosterone-induced effects in vitro and to enlarge accessory sex organs in castrated male rats. This study was designed to evaluate the effects of TCC on intact age-matched male rats and on human prostate LNCaP and C4–2B cells. Seven-week-old male Sprague-Dawley rats received either a normal diet or a diet supplemented with TCC (0.25% in diet) for 10 days. Triclocarban induced hyperplasia of accessory sex organs in the absence of significant qualitative histological changes. Serum luteinizing hormone (LH) and testosterone were not significantly altered by TCC treatment. In prostate cancer-derived LNCaP and C4–2B cells, TCC potentiated androgen actions via androgen receptor-dependent actions. In conclusion, TCC significantly affects intact male reproductive organs and potentiates androgen effects in prostate cancer cells.

Triclocarban (TCC) toxicity and bioaccumulation data are primarily limited to direct human and animal dermal exposures, animal ingestion exposures to neat and feed-spiked TCC, and/or aquatic organism exposures. Three non-human, terrestrial organism groups anticipated to be the most highly exposed to land-applied, biosolids-borne TCC are soil microbes, earthworms, and plants. The three ecological receptors are expected to be at particular risk due to unique modes of exposure (e.g. constant, direct contact with soil; uptake of amended soil and pore water), inherently greater sensitivity to environmental contaminants (e.g. increased body burdens, permeable membranes), and susceptibility to minute changes in the soil environment. The toxicities of biosolids-borne TCC to Eisenia fetida earthworms and soil microbial communities were characterized using adaptations of the USEPA Office of Prevention, Pesticides, and Toxic Substances (OPPTS) Guidelines 850.6200 (Earthworm Subchronic Toxicity Test) and 850.5100 (Soil Microbial Community Toxicity Test), respectively. The resultant calculated TCC LC50 value for E. fetida was 40 mg TCC kg amended fine sand � 1 . Biosolids-borne TCC in an amended fine sand had no significant effect on soil microbial community respiration, ammonification, or nitrification. Bioaccumulation of biosolids-borne TCC by E. fetida and Paspulum notatum was measured to characterize potential biosolidsborne TCC movement through the food chain. Dry-weight TCC bioaccumulation factor (BAF) values in E. fetida and P. notatum ranged from 5.2 – 18 and 0.00041 – 0.007 (gsoil g � 1 tissue ), respectively.

Triclocarban (TCC) is an antimicrobial additive of personal care products that is only partially degraded during wastewater treatment. Bacteria responsible for its transformation are unknown. We obtained wastewater bacteria capable of using as the sole carbon source TCC or its non-chlorinated analog, carbanilide (NCC). Enrichments established using activated sludge amended with TCC and NCC, respectively, were maintained for one year through successive transfers. Enrichments displayed exponential growth after two weeks, reaching stationary phase after one month. The NCC enrichment was shown to accumulate aniline. Denaturing gradient gel electrophoresis of amplified 16S rRNA genes indicated markedly reduced community richness compared to the inoculum and a single, prominent taxonomic unit emerged in both chlorinated and non-chlorinated carbanilide enrichment cultures. Cloned 16S rRNA genes showed both enrichments were dominated by a single genotype related to uncharacterized organisms within the Alcaligenaceae. Of ~30 sequences from each enrichment, no other organisms were detected in the TCC enrichment while a small, flanking community of Alpha Proteobacteria was detected in the NCC enrichment. Study results demonstrate that growth of wastewater bacteria on TCC and its lower chlorinated analog can be linked to bacteria within the family Alcaligenaceae. These organisms are promising agents for the bioremediation of hazardous phenylurea pollutants.

Triclocarban (3,4,4′-trichlorocarbanilide, TCC) is a widely used antibacterial agent in personal care products and is frequently detected as an environmental pollutant in waste waters and surface waters. In this study, we report novel reactive metabolites potentially formed during biotransformation of TCC. The oxidative metabolism of TCC has been predicted using an electrochemical cell coupled online to liquid chromatography and electrospray ionization mass spectrometry. The electrochemical oxidation unveils the fact that hydroxylated metabolites of TCC may form reactive quinone imines. Moreover, a so-far unknown dechlorinated and hydroxylated TCC metabolite has been identified. The results were confirmed by in vitro studies with human and rat liver microsomes. The reactivity of the newly discovered quinone imines was demonstrated by their covalent binding to glutathione and macromolecules, using β-lactoglobulin A as a model protein. The results regarding the capability of the electrochemical cell to mimic the oxidative metabolism of TCC are discussed. Moreover, the occurrence of reactive metabolites is compared with findings from earlier in vivo studies and their relevance in vivo is argued.

This study evaluated the leaching potential of the antimicrobials triclocarban (TCC) and triclosan (TCS) in three agricultural soils using a simple model based on biodegradation and adsorption. The antimicrobials were added to the soils at two moisture levels (10% or 15% w/w) to achieve initial concentrations of 0.05, 0.2, or 2 mg kg(-1). The low concentrations (0.05, 0.2 mg kg(-1)) are more representative of field concentrations, important because previous studies have typically focused on higher initial concentrations. After 100 d, significant residuals of both TCC and TCS occurred under all conditions and first-order degradation half-lives indicated TCC was more resistant to biodegradation. The estimated K(d) and K(oc) values were 193-296 L kg(-1) and 18175-33991 L kg(-1) for TCC and 33-55 L kg(-1) and 3968-6310 L kg(-1) for TCS. The resulting leaching models indicated these chemicals have a very low leaching potential and are thus unlikely to contaminate groundwater.

We investigated two commonly used antimicrobial agents triclosan (TCS) and triclocarban (TCC) in the Pearl River system in China (i.e., Liuxi, Zhujiang and Shijing Rivers) and four sewage effluents during dry and wet seasons. The median values for TCS and TCC were the highest in the surface water and sediments of the Shijing River, followed by the Zhujiang River and Liuxi River. Screening level risk assessment using the risk quotient (RQ) method showed that TCS and TCC in surface water posed median risks in the Zhujiang and Liuxi Rivers (RQs: 0.28-0.62 for TCS, and 0.15-0.80 for TCC) and high risks in the Shijing River (RQs: 5.15-9.55 for TCS, and 3.32-5.83 for TCC). Higher risks (RQs: 3.63-28.47 for TCS, and 3.13-24.54 for TCC) were found in the sediments than in surface water of the Pearl River system. The four sewage effluents and Shijing River as well as other urban streams in Guangzhou metropolitan area were identified as the sources of the two compounds in the main river Zhujiang River. The mass inventories of TCS and TCC in the Pearl River system indicate that the sediments are not only an important sink but also a potential source for the two compounds in surface water.

Propellants containing nitrate esters need stabilizers to avoid early decomposition or even explosion during storage. Newly prepared malonanilides M1-M5 were tested in stabilizing double-base propellants (DBPs). Their stabilization was compared with the effect of classical stabilizer N,N'-diethyldiphenyl urea (C1) using both practical thermal stability tests (qualitative and quantitative tests) and theoretical molecular orbital calculations. This research shows that the new stabilizers (malonanilides) have good stabilizing effects. Some of malonanilides e.g. (M5) and (M2) have higher stability effects. Different mechanisms were suggested to explain the role of different stabilizers. Molecular orbital calculations using the semi-empirical program AM1 are performed on the new and classical stabilizers. Correlation was made between the volume (ml) of NO(x), weight loss (wt%), other thermal analyses data, calculated thermodynamic parameters like activation energy (E(a), kJ mol(-1)) of the decomposed propellant samples containing different stabilizers and some of their calculated properties such as HOMO, LUMO energies, the charge distribution and the pi-bond order. The stabilization effect decreases with the increase in HOMO energy. The correlation between the net charge and parameters measured for the stabilization effect shows good accordance. Comparison of the results obtained show that the high electron charge on N atom of the stabilizers and on its benzene ring is the most important factor, but not the only factor governing the stabilization effect of the stabilizers.

In this work, a new approach for the isolation of nitrocellulose from smokeless gunpowders has been developed. A multistep solvent extraction method was needed to purify nitrocellulose contained in gunpowders. For single-base or double-base gunpowders six consecutive solvent extractions were selected: three extractions with methanol (to remove nitroglycerin, 2,4-dinitrotoluene, ethyl-centralite, diphenylamine, and diphenylamine derivatives); one extraction with dichloromethane (to remove colorants and plasticizers of organic nature); one extraction with methanol (to facilitate a final polar extraction); and one extraction with water (to remove ionic components) were necessary at 35 degrees C. For the triple-base gunpowder studied, eight solvent extractions were needed due to a high concentration of the water-soluble nitroguanidine was present. In addition to the same five initial phases used for the single-base and double-base gunpowders, three water extraction phases at a higher temperature (75 degrees C instead of 35 degrees C) were also needed. A final step to solubilize nitrocellulose in methyl ethyl ketone was used to remove inert components (mainly graphite). Nitrocellulose isolated from these propellants was characterized by Fourier-Transformed Infrared Spectroscopy (FTIR spectroscopy). The same FTIR spectra were observed for nitrocelluloses isolated from different types of gunpowders. A comparison of FTIR spectra of nitrocellulose samples of different nitration degree evidenced that the bands regions most affected by this factor were: 3600-3400cm(-1), corresponding to the stretching vibrations of residual hydroxyl groups; 1200-1000cm(-1), attributed to the valence vibrations nuCO of the glucopyranose cycle; and 750-690cm(-1), assigned to vibrations of the nitrate group. In both cases, the bands appearing in these regions were more pronounced in the spectra of nitrocellulose samples of low nitration degree.

Triclocarban (TCC) is an active ingredient in antibacterial bar soaps, a common constituent of domestic wastewater, and the subject of recent criticism by consumer advocate groups and academic researchers alike. Activated sludge treatment readily removes TCC from the liquid waste stream and concentrates the antimicrobial in the solid fraction, which is often processed to produce biosolids intended for land application. Greater than half of the biosolids generated in the US are land-applied, resulting in a systematic release of biosolids-borne TCC into the terrestrial and, potentially, the aquatic environment. Multiple data gaps in the TCC literature (including basic physicochemical properties and biosolids concentrations) prevent an accurate, quantitative risk assessment of biosolids-borne TCC. We utilized the USEPA Office of Prevention, Pesticides, and Toxic Substances (OPPTS) harmonized test guidelines to measure TCC solubility and log K ow values as 0.045 mg L − 1 and 3.5, respectively. The measured physicochemical 2 properties differed from computer model predictions. The mean concentration of TCC in 23 biosolids representative of multiple sludge processing methods was 19 ± 11 mg kg − 1 .

Triclocarban (TCC) is an antibacterial compound commonly detected in biosolids at parts-per-million concentrations. Approximately half of the biosolids produced in the United States are land-applied, resulting in a systematic release of TCC into the soil environment. The extent of biosolids-borne TCC environmental transport and potential human/ecological exposures will be greatly affected by its bioavailability and the rate of degradation in amended soils. To investigate these factors, radiolabeled TCC ((14)C-TCC) was incorporated into anaerobically digested biosolids, amended to two soils, and incubated under aerobic conditions. The evolution of (14)CO2 (biodegradation) and changes in chemical extractability (bioavailability) was measured over time. Water extractable TCC over the study period was low and significantly decreased over the first 3 weeks of the study (from 14% to 4% in a fine sand soil and from 3 to

A novel dispersive liquid-phase microextraction method without dispersive solvents has been developed for the enrichment and sensitive determination of triclosan and triclocarban in environmental water samples prior to HPLC-ESI-MS/MS. This method used only green solvent 1-hexyl-3-methylimidazolium hexafluorophosphate as extraction solvent and overcame the demerits of the use of toxic solvents and the instability of the suspending drop in single drop liquid-phase microextraction. Important factors that may influence the enrichment efficiencies, such as volume of ionic liquid, pH of solutions, extraction time, centrifuging time and temperature, were systematically investigated and optimized. Under optimum conditions, linearity of the method was observed in the range of 0.1-20 microg/L for triclocarban and 0.5-100 microg/L for triclosan, respectively, with adequate correlation coefficients (R>0.9990). The proposed method has been found to have excellent detection sensitivity with LODs of 0.04 and 0.3 microg/L, and precisions of 4.7 and 6.0% (RSDs, n=5) for triclocarban and triclosan, respectively. This method has been successfully applied to analyze real water samples and satisfactory results were achieved.

The interaction of drugs and non-therapeutic xenobiotics constitutes a central role in human health risk assessment. Still, available data are rare. Two different models have been established to predict mixture toxicity from single dose data, namely, the concentration addition (CA) and independent action (IA) model. However, chemicals can also act synergistic or antagonistic or in dose level deviation, or in a dose ratio dependent deviation. In the present study we used the MIXTOX model (EU project ENV4-CT97-0507), which incorporates these algorithms, to assess effects of the binary mixtures in the human hepatoma cell line HepG2. These cells possess a liver-like enzyme pattern and a variety of xenobiotic-metabolizing enzymes (phases I and II). We tested binary mixtures of the metal nickel, the anti-inflammatory drug diclofenac, and the antibiotic agent irgasan and compared the experimental data to the mathematical models. Cell viability was determined by three different methods the MTT-, AlamarBlue(R) and NRU assay. The compounds were tested separately and in combinations. We could show that the metal nickel is the dominant component in the mixture, affecting an antagonism at low-dose levels and a synergism at high-dose levels in combination with diclofenac or irgasan, when using the NRU and the AlamarBlue assay. The dose-response surface of irgasan and diclofenac indicated a concentration addition. The experimental data could be described by the algorithms with a regression of up to 90%, revealing the HepG2 cell line and the MIXTOX model as valuable tool for risk assessment of binary mixtures for cytotoxic endpoints. However the model failed to predict a specific mode of action, the CYP1A1 enzyme activity.

Recent research has indicated that the antimicrobial chemical triclocarban (TCC) represents a new type of endocrine disruptor, amplifying the transcriptional activity of steroid hormones and their receptors while itself exhibiting little affinity for these receptors. The effects of TCC were studied in the freshwater mudsnail Potamopyrgus antipodarum. Specimens were exposed to concentrations ranging from 0.05 to 10.5 microg/L dissolved TCC and were removed and dissected, and embryos contained within the brood pouch were counted and classified as shelled or unshelled after two and four weeks of exposure. After four weeks, environmentally relevant TCC concentrations of 1.6 to 10.5 microg/L resulted in statistically significant increases in the number of unshelled embryos, whereas 0.2, 1.6, and 10.5 microg/L exposures significantly increased numbers of shelled embryos. The lowest observed effect concentration (LOEC) was 0.2 microg/L, the no observed effect concentration (NOEC) was 0.05 microg/L; the 10% effective concentration (EC10) and the median effective concentration (EC50) for unshelled effects were 0.5 microg/L and 2.5 microg/L, respectively. Given the widespread occurrence of TCC in the environment and the effects shown at environmentally relevant concentrations, these results indicate that TCC may be causing reproductive effects in the environment. Furthermore, the present study indicates that environmental risk from a new class of endocrine-disrupting chemicals (EDCs) is both qualitatively and quantitatively similar to risk from existing classes of EDCs.

Triclocarban is an antimicrobial and antibacterial agent found in personal care products and subsequently is a prevalent wastewater contaminant. A quantitative method was developed for the analysis of triclocarban in wastewater effluents using stir bar sorptive extraction–liquid desorption (SBSE–LD) followed by liquid chromatography–tandem mass spectrometry (LC–MS/MS) by means of an electrospray interface. A stir bar coated with polydimethylsiloxane (PDMS) is placed within a vial containing wastewater effluent and is stirred for an hour at room temperature. The PDMS stir bar is then placed in a LC vial containing methanol and is desorbed in a sonicator bath. The methanol is evaporated to dryness and reconstituted in 75% methanol. Spike and recovery experiments in groundwater that did not contain native concentrations of triclocarban were performed at 0.5 μg/L and were 93 ± 8%. Recoveries in wastewater effluent that were corrected for the background levels of triclocarban were 92 ± 2% and 96 ± 5%, respectively, when spiked with 0.5 and 5 μg/L of triclocarban. The precision of the method as indicated by the relative standard error was 2%. The limit of quantitation was 10 ng/L. The SBSE–LD–LC/MS/MS method was applied to wastewater effluent samples collected from northeast Ohio. Triclocarban was quantitated in all five effluent samples, and its concentration ranged from 50 to 330 ng/L. The described method demonstrates a simple, green, low-sample volume, yet, sensitive method to measure triclocarban in aqueous matrices.

Microglial cells play an important role during neuroinflammation in the central nervous system. Among other factors, activated microglia produce nitric oxide (NO), which is toxic to neurons and excessive microglial activation and NO production contribute to the pathology of neurodegenerative diseases. Chloride channels have previously been shown to participate in microglial activation. Here we investigate the effects of established chloride channel blockers with different chemical structures on interferon-gamma (IFNgamma)-induced activation of the murine microglial cell line, BV2. IFNgamma-induced NO production was effectively reduced by NPPB, IAA-94, tamoxifen, NS3728 and NS1652, with NS1652 being the most potent. In contrast, DIDS reduced NO production only at cytotoxic concentrations. Furthermore, NS1652 reduced the protein and mRNA levels of inducible nitric oxide synthase (iNOS), without altering STAT1 phosphorylation. These observations suggest a microglial chloride conductance as a critical permissive factor downstream in the IFNgamma-induced iNOS cascade. The nature of the underlying channel is unknown, but the pharmacological profile appears incompatible with the involvement of the volume activated anion conductance (VRAC).

Triclosan and triclocarban are antibacterial agents that are widely used in numerous personal care products. Limited information is available on their environmental behavior in soils and soils land applied with wastewaters and biosolids. In this study, laboratory experiments were performed to investigate their adsorption and degradation in soils. Both antibacterial agents adsorbed strongly to the sandy loam and silty clay soils with and without addition of biosolids, with distribution coefficients (K(d)) ranging from 178 to 264 L kg(-1) for triclosan and from 763 to 1187 L kg(-1) for triclocarban. Sorption of triclosan decreased with increase in soil pH from 4 to 8, whereas triclocarban sorption showed no effect within the tested pH range. Competitive sorption was observed when triclosan and triclocarban coexisted, but the cosolute effect was concentration dependent. Biosolids amendment increased the sorption of triclosan and triclocarban, likely due to the addition of soil organic matter, but displayed no significant effect on degradation.

The occurrence of the antimicrobials triclocarban (TCC) and triclosan (TCS) was investigated in agricultural soils following land application of biosolids using liquid chromatography-tandem mass spectrometry (LC-MS-MS) with negative ion multimode ionization. The method detection limits were 0.58 ng TCC/g soil, 3.08 ng TCC/g biosolids, 0.05 ng TCS/g soil and 0.11 ng TCS/g biosolids and the average recovery from all of the sample matrices was95%. Antimicrobial concentrations in biosolids from three Michigan wastewater treatment plants (WWTPs) ranged from 4890 to 9280 ng/g, and from 90 to 7060 ng/g, for TCC and TCS respectively. Antimicrobial analysis of soil samples, collected over two years, from ten agricultural sites previously amended with biosolids, indicated TCC was present at higher concentrations (1.24-7.01 ng/g and 1.20-65.10 ng/g in 2007 and 2008) compared to TCS (0.16-1.02 ng/g and from the method detection limit,0.05-0.28 ng/g in 2007 and 2008). Soil antimicrobial concentrations could not be correlated to any soil characteristic, or to the time of last biosolids application, which occurred in either 2003, 2004 or 2007. To our knowledge, our data represent the first report of TCC, and the first comparison of TCC and TCS concentrations, in biosolids-amended agricultural soils. Such information is important because approximately 50% of US biosolids are land applied, therefore, any downstream effects of either antimicrobial are likely to be widespread.

An attractive target for therapeutic intervention is constitutively activated, mutant FLT3, which is expressed in a subpopulation of patients with acute myelocyic leukemia (AML) and is generally a poor prognostic indicator in patients under the age of 65 years. PKC412 is one of several mutant FLT3 inhibitors that is undergoing clinical testing, and which is currently in late-stage clinical trials. However, the discovery of drug-resistant leukemic blast cells in PKC412-treated patients with AML has prompted the search for novel, structurally diverse FLT3 inhibitors that could be alternatively used to override drug resistance. Here, we report the potent and selective antiproliferative effects of the novel mutant FLT3 inhibitor NVP-AST487 on primary patient cells and cell lines expressing FLT3-ITD or FLT3 kinase domain point mutants. NVP-AST487, which selectively targets mutant FLT3 protein kinase activity, is also shown to override PKC412 resistance in vitro, and has significant antileukemic activity in an in vivo model of FLT3-ITD+ leukemia. Finally, the combination of NVP-AST487 with standard chemotherapeutic agents leads to enhanced inhibition of proliferation of mutant FLT3-expressing cells. Thus, we present a novel class of FLT3 inhibitors that displays high selectivity and potency toward FLT3 as a molecular target, and which could potentially be used to override drug resistance in AML.

Background Concerns have been raised about the biological and toxicologic effects of the antimicrobials triclocarban (TCC) and triclosan (TCS) in personal care products. Few studies have evaluated their biological activities in mammalian cells to assess their potential for adverse effects. Objectives In this study, we assessed the activity of TCC, its analogs, and TCS in in vitro nuclear-receptor–responsive and calcium signaling bioassays. Materials and methods We determined the biological activities of the compounds in in vitro, cell-based, and nuclear-receptor–responsive bioassays for receptors for aryl hydrocarbon (AhR), estrogen (ER), androgen (AR), and ryanodine (RyR1). Results Some carbanilide compounds, including TCC (1–10 μM), enhanced estradiol (E2)-dependent or testosterone-dependent activation of ER- and AR-responsive gene expression up to 2.5-fold but exhibited little or no agonistic activity alone. Some carbanilides and TCS exhibited weak agonistic and/or antagonistic activity in the AhR-responsive bioassay. TCS exhibited antagonistic activity in both ER- and AR-responsive bioassays. TCS (0.1–10 μM) significantly enhanced the binding of [3H]ryanodine to RyR1 and caused elevation of resting cytosolic [Ca2+] in primary skeletal myotubes, but carbanilides had no effect. Conclusions Carbanilides, including TCC, enhanced hormone-dependent induction of ER- and AR-dependent gene expression but had little agonist activity, suggesting a new mechanism of action of endocrine-disrupting compounds. TCS, structurally similar to noncoplanar ortho-substituted poly-chlorinated biphenyls, exhibited weak AhR activity but interacted with RyR1 and stimulated Ca2+ mobilization. These observations have potential implications for human and animal health. Further investigations are needed into the biological and toxicologic effects of TCC, its analogs, and TCS.

Alkyl nitrates and very short-lived halocarbon species are important atmospheric trace gas species that are present in the low to sub parts per trillion concentration range. This presents an analytical challenge for their detection and quantification that requires instrumentation with high sensitivity and selectivity. In this paper, we present a new in situ gas chromatograph negative ion chemical ionization mass spectrometer (GC/NICI-MS) coupled to a non-cryogen sample pre-concentration system. This instrument, with detection limits of

A simple and easy-to-build high-throughput analysis system was constructed. The system consisted of three major components: (1) a multichannel device with 16 parallel capillaries, (2) a desorption electrospray ionization (DESI) source, and (3) a linear ion trap mass spectrometer. When analyses were performed, the multichannel device was moved horizontally on a translation stage controlled by a step motor. Our design expands the functions of DESI, in which the liquid sample in capillary was driven out by the nebulizing gas, ionized, and then transferred to a mass spectrometer. To assess the high-throughput performance of the system, 5 mg/L 1,3-diethyl-1,3-diphenylurea (DDU) solution and 10 mg/L angiotensin I solution were alternatively loaded into the reservoirs and capillaries in the multichannel device. Results indicated that analyses of the all the samples in 16 capillaries were completed within 1.6 min, which means a throughput of 600 samples/h. Reactive DESI experiment was also successfully performed with this system to show the feasibility of online derivatization. The relative standard deviations for a single capillary and five identical capillaries were 7.6 (n = 16) and 12.3%, respectively. Linear relative abundance response was achieved for DDU (r = 0.9971).

Triclocarban and triclosan are two antimicrobial agents widely used in many personal care products. Their biodegradation behaviour in soil was investigated by laboratory degradation experiments and environmental fate modelling. Quantitative structure-activity relationship (QSAR) analyses showed that triclocarban and triclosan had a tendency to partition into soil or sediment in the environment. Fate modelling suggests that either triclocarban or triclosan "does not degrade fast" with its primary biodegradation half-life of "weeks" and ultimate biodegradation half-life of "months". Laboratory experiments showed that triclocarban and triclosan were degraded in the aerobic soil with half-life of 108 days and 18 days, respectively. No negative effect of these two antimicrobial agents on soil microbial activity was observed in the aerobic soil samples during the experiments. But these two compounds persisted in the anaerobic soil within 70 days of the experimental period.

The expression of the H-ras oncogene increases the migratory activity of many cell types and thereby contributes to the metastatic behavior of tumor cells. Other studies point to an involvement of volume-activated anion channels (VRAC) in (tumor) cell migration. In this paper, we tested whether VRACs are required for the stimulation of cell migration upon expression of the H-ras oncogene. We compared VRAC activation and migration of wild-type and H-ras-transformed NIH3T3 fibroblasts by means of patch-clamp techniques and time-lapse video microscopy. Both cell types achieve the same degree of VRAC activation upon maximal stimulation, induced by reducing extracellular osmolarity from 300 to 190 mOsm/l. However, upon physiologically relevant reductions in extracellular osmolarity (275 mOsm/l), the level of VRAC activation is almost three times higher in H-ras-transformed compared to wild-type fibroblasts. This increase in VRAC sensitivity is accompanied by increased migratory activity of H-ras fibroblasts. Moreover, the high-affinity VRAC blocker NS3728 inhibits migration of H-ras fibroblasts dose-dependently by up to about 60%, whereas migration of wild-type fibroblasts is reduced by only about 35%. Consistent with higher VRAC activity in H-ras than in wild-type fibroblasts, more VRAC blocker is needed to achieve a comparable degree of inhibition of migration. We suggest that H-ras modulates the volume set point of VRAC and thus facilitates transient changes of cell volume required for faster cell migration.

A sensitive and accurate method was developed for the simultaneous determination of triclosan and triclocarban in sludge and treated biosolids from municipal wastewater treatment plants (WWTPs). The methods involved extraction by pressurized liquid extraction (PLE), followed by sample clean-up on a Oasis HLB solid phase extraction (SPE) cartridge and analysis by liquid chromatography with electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS). Accurate quantification was achieved by isotope dilution using stable isotopes of triclosan and triclocarban as internal standards. Matrix effects (ME) in samples of spiked biosolids (n=5) were evaluated by a standard addition method, and these analyses indicated mean ME values of 79.7+/-6.7 and 100.5+/-8.4% for triclosan and triclocarban, respectively; indicating that the sample clean-up method effectively removed interferences. The mean recoveries from the spiked biosolids sample were 97.7+/-6.2 and 98.3+/-5 for triclosan and triclocarban, respectively, and the limits of detection (LOD) were 1.5 and 0.2 ng/g (d.w.) for triclosan and triclocarban, respectively. The method was applied to the analysis of triclosan and triclocarban in samples of activated sludge and treated biosolids collected from three WWTPs in Ontario, Canada. These preliminary results indicate that triclosan and triclocarban co-occur in municipal sludge and treated biosolids at concentrations ranging from 0.62 to 11.55 microg/g dry weight for triclosan, and from 2.17 to 5.97 microg/g dry weight for triclocarban.

accepted in Journal of Chemical Information Modeling; Though different species of the genus Plasmodium may be responsible for malaria, the variant caused by P. falciparum is often very dangerous and even fatal if untreated. Hemoglobin degradation is one of the key metabolic processes for the survival of the Plasmodium parasite in its host. Plasmepsins, a family of aspartic proteases encoded by the Plasmodium genome, play a prominent role in host hemoglobin cleavage. In this paper we demonstrate the use of virtual screening, in particular molecular docking, employed at a very large scale to identify novel inhibitors for plasmepsins II and IV. A large grid infrastructure, the EGEE grid, was used to address the problem of large computation resources required for docking hundreds of thousands of chemical compounds on different plasmepsin targets of P. falciparum. A large compound library of about 1 million chemical compounds was docked on 5 different targets of plasmepsins using two different docking software, namely FlexX and AutoDock. Several strategies were employed to analyze the results of this virtual screening approach including docking scores, ideal binding modes, and interactions to key residues of the protein. Three different classes of structures with thiourea, diphenylurea, and guanidino scaffolds were identified to be promising hits. While the identification of diphenylurea compounds is in accordance with the literature and thus provides a sort of "positive control", the identification of novel compounds with a guanidino scaffold proves that high throughput docking can be effectively used to identify novel potential inhibitors of P. falciparum plasmepsins. Thus, with the work presented here, we do not only demonstrate the relevance of computational grids in drug discovery but also identify several promising small molecules which have the potential to serve as candidate inhibitors for P. falciparum plasmepsins. With the use of the EGEE grid infrastructure for the virtual screening campaign against the malaria causing parasite P. falciparum we have demonstrated that resource sharing on an eScience infrastructure such as EGEE provides a new model for doing collaborative research to fight diseases of the poor

Algae comprise the greatest abundance of plant biomass in aquatic environments and are a logical choice for aquatic toxicological studies, yet have been underutilized in this capacity. The lipid content of many algal species provides a point of entry for trophic transfer of lipophilic organic contaminants. Triclosan (TCS) and triclocarban (TCC), widely used antimicrobial agents found in numerous consumer products, are incompletely removed by wastewater treatment plant (WWTP) processing. Methyl-triclosan (M-TCS) is a metabolite of TCS more lipophilic than the parent compound. The focus of this study was to quantify algal bioaccumulation factors (BAFs) for TCS, M-TCS, and TCC in Pecan Creek, the receiving stream for the City of Denton, Texas WWTP. The complex algal compartment was field identified for collection and verified by laboratory microscopic description as being comprised of mostly filamentous algae (Cladophora spp.) and varying inconsequential levels of epiphytic diatoms and biofilm. Algae and water column samples were collected from the WWTP outfall, an upstream site, and two downstream sites and analysed by isotope dilution gas chromatography/mass spectrometry (GC/MS) for TCS and M-TCS and liquid chromatography/mass spectrometry (LC/MS) for TCC. TCS, M-TCS, and TCC in Pecan Creek water samples taken at and downstream from the WWTP were at low ppt concentrations of 50-200 ng l(-1) and were elevated to low ppb concentrations of 50-400 ng g(-1) fresh weight in algae collected from these stations. The resulting BAFs were approximately three orders of magnitude. TCS, M-TCS and TCC appear to be good candidate marker compounds for evaluation of environmental distribution of trace WWTP contaminants. Residue analysis of filamentous algal species typically occurring in receiving streams below WWTP discharges is a readily obtained indicator of the relative bioaccumulative potential of these trace contaminants.

Background Prior studies suggest associations between fetal exposure to antimicrobial and paraben compounds with adverse reproductive outcomes, mainly in animal models. We have previously reported elevated levels of these compounds for a cohort of mothers and neonates. Objective We examined the relationship between human exposure to parabens and antimicrobial compounds and birth outcomes including birth weight, body length and head size, and gestational age at birth. Methods Maternal third trimester urinary and umbilical cord blood plasma concentrations of methylparaben (MePB), ethylparaben (EtPB), propylparaben (PrPB), butylparaben (BuPB), benzylparaben (BePB), triclosan (2,4,4′-trichloro-2′-hydroxydiphenyl ether or TCS) and triclocarban (1-(4-chlorophenyl)-3-(3,4-dichlorophenyl) urea or TCC), were measured in 185 mothers and 34 paired singleton neonates in New York, 2007–2009. Results In regression models adjusting for confounders, adverse exposure-outcome associations observed included increased odds of PTB (BuPB), decreased gestational age at birth (BuPB and TCC) and birth weight (BuPB), decreased body length (PrPB) and protective effects on PTB (BePB) and LBW (3′-Cl-TCC) ( p Conclusions This study provides the first evidence of associations between antimicrobials and potential adverse birth outcomes in neonates. Findings are consistent with animal data suggesting endocrine-disrupting potential resulting in developmental and reproductive toxicity.

Triclosan, triclocarban, 2,2',4,4'-tetrabromodiphenyl ether (BDE-47), and bisphenol A (BPA) have been reported to disturb thyroid hormone (TH) homeostasis. We have examined the effects of these chemicals on sodium/iodide symporter (NIS)-mediated iodide uptake and the expression of genes involved in TH synthesis in rat thyroid follicular FRTL-5 cells, and on the activity of thyroid peroxidase (TPO) using rat thyroid microsomes. All four chemicals inhibited NIS-mediated iodide uptake in a concentration-dependent manner. A decrease in the iodide uptake was also observed in the absence of sodium iodide. Kinetic studies showed that all four chemicals were non-competitive inhibitors of NIS, with the order of Ki values being triclosantriclocarbanBDE-47BPA. The transcriptional expression of three genes involved in TH synthesis, Slc5a5, Tpo, and Tgo, and three thyroid transcription factor genes, Pax8, Foxe1, and Nkx2-1, was examined using quantitative real-time PCR. No significant changes in the expression of any genes were observed with triclosan or triclocarban. BDE-47 decreased the level of Tpo, while BPA altered the expression of all six genes. Triclosan and triclocarban inhibited the activity of TPO at 166 and300 μM, respectively. Neither BDE-47 nor BPA affected TPO activity. In conclusion, triclosan, triclocarban, BDE-47, and BPA inhibited iodide uptake, but had differential effects on the expression of TH synthesis-related genes and the activity of TPO.

A sensitive, competitive indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of triclocarban (TCC) in waters and sediments. Haptens were synthesized by derivatizing the paraposition of a phenyl moiety of TCC. The synthesized hapten was then coupled to bovine thyroglobulin to be used as an immunogen, based on which, a high affinity monoclonal antibody 4D5 was produced with the hybridoma technique. Under the optimized conditions, using the monoclonal antibody, excellent performances of the assay were obtained: satisfactory sensitivity (IC50 (50% inhibition concentration) value, 0.43 ng/mL; limit of detection, 0.05 ng/mL); good linear range (0.05–10 ng/mL); and satisfactory accuracy (recoveries 70.7–107% in waters; 74.8–98.3% in sediments). Furthermore, TCC was found with the concentration ranging from not detected to 422.12 ng/L in waters and from 6.68 ng/g to 78.67 ng/g in sediments in Yunliang River, Ancient Canal and Hongqiao Port in Zhenjiang City. In conclusion ELISA could be applied for monitoring TCC in aquatic environments.

Triclosan (TCS) and triclocarban (TCC) exposures are highly concerned due to their suspected endocrine-disrupting effects. The present study investigated TCS and TCC exposure levels in the general Chinese population by biomonitoring human urine and nail samples. TCS (69-80 %) and TCC (99-100 %) were frequently detected, which demonstrates that the general Chinese population has extensive exposure to these chemicals. The geometric mean (GM) urinary concentrations were 0.40 μg/g creatinine (creat), 95 % confidence interval (CI) 0.30-0.56, for TCS and 0.40 μg/g creat, 95 % CI 0.29-0.56, for TCC. On the other hand, the GM levels of TCS and TCC were 13.57 (5.67 μg/kg) and 84.66 μg/kg (41.50 μg/kg) in fingernail (toenail) samples, respectively, indicating that the levels in fingernails were approximately twice as high as those in toenails. Pearson's correlation coefficients between the urine and fingernail (toenail) samples were 0.715 (0.614) for TCS and 0.829 (0.812) for TCC. These data suggest that nail samples can be applied to the biomonitoring for TCS and TCC in the general population. We observed that the levels of both chemicals were higher in females than in males for urine and fingernail samples, but no significant differences were found between different genders for either compound in toenails. Nineteen- to 29-year-olds had the highest TCS levels in their nail samples, whereas TCC levels did not differ with regard to age. Region of residence significantly influenced TCS and TCC concentrations in the three biological matrices measured.

An analytical method was developed to simultaneously detect triclosan (TCS) and triclocarban (TCC) in human breast milk using solid-phase extraction (SPE) with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).Samples were extracted by acetonitrile and purified with C -18 SPE cartridge after enzymolysis with β-glucuronidase/arylsulfatase. The chromatographic separation was performed on a Waters ACQUITY UPLC™ HSS T3 column (100 mm x 2. 1 mm, 1. 8 µm) with gradient elution using methanol and water at a flow rate of 0. 3 ml/min. The target analytes were assayed by triple quadrupole mass spectrometer operating in the negative ion mode. Quantification was performed by isotopic internal standard calibration.Satisfactory linearity (r20. 999) was obtained over the range of 0. 2 - 20. 0 µg/L and 0. 02 - 2. 0 µg/L for triclosan and triclocarban, respectively, with the limits of quantifications (LOQs) of 0. 41 and 0. 03 µg/kg. Average recoveries of two target compounds (spiked at three concentration levels) ranged from 100. 2% to 119. 3%, with the relative standard deviations (RSDs) between 5. 91% and 11. 31% (n =6). Twenty-five real samples (n = 25) were detected containing TCS and TCC at concentrations ofLOQ - 0. 77 µg/kg andLOQ - 4. 28 µg/kg, respectively.Due to its high sensitivity and good reproductivity, this method can be applied to analyze TCS and TCC in human breast milk.