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Sample records for 5-diphenyl-tetrazolium bromide mtt

  1. Ammonia-containing dimethyl sulfoxide: an improved solvent for the dissolution of formazan crystals in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay.

    PubMed

    Wang, Hengwei; Wang, Fengqing; Tao, Xinyi; Cheng, Hairong

    2012-02-01

    To reduce interference with the dissolution of formazan crystals in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, we conducted a systematic investigation to study the effects of various buffers, HCl, NaOH, and ammonia. As a result, we identified an improved solvent, alkaline dimethyl sulfoxide (DMSO) containing 8 to 800 mM ammonia, which could dissolve formazan crystals in approximately 10 min so as to give a stable spectrum by eliminating buffering effects of the residual medium.

  2. Effects of the pH dependence of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide-formazan absorption on chemosensitivity determined by a novel tetrazolium-based assay.

    PubMed

    Plumb, J A; Milroy, R; Kaye, S B

    1989-08-15

    The tetrazolium dye, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), is reduced by live but not dead cells, and this reaction is used as the end point in a rapid drug-screening assay. It can also be used for accurate determinations of drug sensitivity but only if a quantitative relationship is established between cell number and MTT-formazan production. We have shown that reduction of MTT to MTT-formazan by cells is dependent on the amount of MTT in the incubation medium. The concentration required to give maximal MTT-formazan production differs widely between cell lines. The absorption spectrum of MTT-formazan varies with cell number and with pH. At a low cell density or a high pH, the absorption maximum is at a wavelength of 560 to 570 nm. However, at a high cell density or a low pH, there are two absorption maxima; one at 510 nm and a second at about 570 nm. Measurements of absorbance at 570 nm underestimate MTT-formazan production and, hence, cell number at high cell densities. This error can result in a 10-fold underestimation of chemosensitivity. Addition of a buffer at pH 10.5 to the solubilized MTT-formazan product can overcome the effects of both cell density and culture medium on the absorption spectrum. Provided that sufficient MTT is used and the pH of the MTT-formazan product is controlled, dye reduction can be used to estimate cell numbers in a simple chemosensitivity assay the results of which agree well with a commonly used clonogenic assay.

  3. Cytotoxic amyloid peptides inhibit cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction by enhancing MTT formazan exocytosis.

    PubMed

    Liu, Y; Schubert, D

    1997-12-01

    Amyloid beta peptide (A beta) neurotoxicity is believed to play a central role in the pathogenesis of Alzheimer's disease. An early indicator of A beta toxicity is the inhibition of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction to MTT formazan, a widely used assay for measuring cell viability. In this report we show that A beta and other cytotoxic amyloid peptides such as human amylin dramatically enhance MTT formazan exocytosis, resulting in the inhibition of cellular MTT reduction. Only the amyloid peptides that are known to be cytotoxic enhanced MTT formazan exocytosis. Basal MTT formazan exocytosis and amyloid peptide-enhanced MTT formazan exocytosis are blocked by several drugs with diverse known effects. These and other data suggest that MTT formazan exocytosis is a multistep process and that cytotoxic amyloid peptides enhance MTT formazan exocytosis through an intracellular signal transduction pathway.

  4. Spectrophotometric quantitation of DNA on blots after ethanol-solubilization of the MTT-formazan from anti-digoxigenin-based detection of nucleic acids.

    PubMed

    Colgan, D J

    1993-01-01

    The tetrazolium salt, 3-(4,5-dimethyl-thiazolyl-2)-2,5-diphenyl tetrazolium bromide (MTT) has recently been established as a substitute for Nitro-blue tetrazolium (NBT) in stain mixtures using antibody-conjugated alkaline phosphatase for the location of proteins on Western blots (Heegaard, 1990). Experiments reported here show that MTT is as sensitive as NBT in digoxigenin-labeled probe localization (on nucleic acid blots) utilizing alkaline-phosphatase-labelled, anti-digoxigenin antibodies. Moreover, as the formazan from MTT is soluble in ethanol, it is shown that spectrophotometric quantitation can be used to estimate the amount of target DNA on dot and Southern blots. For dot blotting, pBR328 was used as the probe and pBR322 as target. For Southern blots, human rDNA was used as the probe and total genomic calf DNA as the target. Staining response was linear over at least six twofold DNA dilutions in both types of blot.

  5. Steroid hormones block amyloid fibril-induced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) formazan exocytosis: relationship to neurotoxicity.

    PubMed

    Liu, Y; Schubert, D

    1998-12-01

    Perhaps the most reproducible early event induced by the interaction of amyloid beta peptide (A beta) with the cell is the inhibition of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. We recently demonstrated that cytotoxic amyloid peptides such as A beta and human amylin inhibit cellular MTT reduction by dramatically enhancing MTT formazan exocytosis. We now show the following: (a) Insulin and glucagon, when converted to fibrils with beta-pleated sheet structure, induce MTT formazan exocytosis that is indistinguishable from that induced by A beta. NAC35, an amyloidogenic fragment of alpha-synuclein (or NACP), also induces MTT formazan exocytosis. (b) All protein fibrils with the beta-pleated sheet structure examined are toxic to rat hippocampal neurons. (c) Many sterol sex hormones (e.g., estradiol and progesterone) block amyloid fibril-enhanced MTT formazan exocytosis as well as MTT formazan exocytosis in control cells by acting at a common late step in the exocytic pathway. Steroids fail, however, to protect hippocampal neurons from acute amyloid fibril toxicity. These findings suggest that the ability to enhance MTT formazan exocytosis and to induce neurotoxicity are common biological activities of protein fibrils with beta-pleated sheet structure but that enhanced MTT formazan exocytosis is not sufficient for acute A beta neurotoxicity.

  6. Liposomes and MTT cell viability assay: an incompatible affair.

    PubMed

    Angius, Fabrizio; Floris, Alice

    2015-03-01

    The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay is commonly used to evaluate the cytotoxicity potential of drugs vehicled by liposomes. However, liposome delivering drugs could produce inconsistent values of MTT absorbance. On the basis of previous experiments demonstrating the MTT affinity for lipid droplets, this paper aims to show that empty-liposomes interfere, per se, on MTT assay due to its lipidic nature. This brings into question the use of MTT testing cytotoxicity when liposomes are involved in delivering drugs.

  7. Exocytosis of MTT formazan could exacerbate cell injury.

    PubMed

    Lü, Lanhai; Zhang, Lihong; Wai, Maria Sen Mun; Yew, David Tai Wai; Xu, Jie

    2012-06-01

    MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] method is one of the most widely used methods to analyze cell proliferation and viability. It is taken up through endocytosis and is reduced by mitochondrial enzymes as well as endosomal/lysosomal compartments, then is transported to cell surfaces to form needle-like MTT formazans; however the effect of MTT itself still remains elusive. Our objective was to investigate the direct effects of MTT on in vitro SH-SY5Y cells. Results showed that the endocytosis of MTT did not cause obvious lesion and induce cell death, but the metabolism and exocytosis of MTT could dramatically damage cells. Our results also indicated that MTT could activate apoptosis related factors such as caspase-8, caspase-3 or accelerate the leakage of cell contents after the appearance of MTT formazan crystals. The present data suggest MTT method should be carefully chosen; otherwise the cell viability would be underestimated and incomparable.

  8. Antiproliferative Effect of Solanum nigrum on Human Leukemic Cell Lines

    PubMed Central

    Gabrani, Reema; Jain, Ramya; Sharma, Anjali; Sarethy, Indira P.; Dang, Shweta; Gupta, S.

    2012-01-01

    Solanum nigrum is used in various traditional medical systems for antiproliferative, antiinflammatory, antiseizure and hepatoprotective activities. We have evaluated organic solvent and aqueous extracts obtained from berries of Solanum nigrum for antiproliferative activity on leukemic cell lines, Jurkat and HL-60 (Human promyelocytic leukemia cells). The cell viability after the treatment with Solanum nigrum extract was measured by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay. Results indicated increased cytotoxicity with increasing extract concentrations. Comparative analysis indicated that 50% inhibitory concentration value of methanol extract is the lowest on both cell lines. PMID:23716874

  9. Amyloid beta protein inhibits cellular MTT reduction not by suppression of mitochondrial succinate dehydrogenase but by acceleration of MTT formazan exocytosis in cultured rat cortical astrocytes.

    PubMed

    Abe, K; Saito, H

    1998-08-01

    Alzheimer's disease amyloid beta protein (Abeta) inhibits cellular reduction of the dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Kaneko et al. have previously hypothesized that Abeta works by suppressing mitochondrial succinate dehydrogenase (SDH), but Liu and Schubert have recently demonstrated that Abeta decreases cellular MTT reduction by accelerating the exocytosis of MTT formazan in neuronal cells. To ask which is the case in astrocytes, we compared the effects of Abeta and 3-nitropropionic acid (3-NP), a specific SDH inhibitor, on MTT reduction in cultured rat cortical astrocytes. Treatment with 3-NP (10 mM) decreased cellular activity of MTT reduction, regardless of the time of incubation with MTT. On the other hand. Abeta-induced inhibition of cellular MTT reduction was dependent on the time of incubation with MTT. The cells treated with Abeta (0.1-1000 nM) exhibited normal capacity for MTT reduction at an early stage of incubation ( < 30 min), but ceased to reduce MTT at the late stage (> 1 h). Microscopic examination revealed that Abeta treatment accelerated the appearance of needle-like MTT formazan crystals at the cell surface. These observations support that Abeta accelerates the exocytosis of MTT formazan in astrocytes. In addition to inhibition of MTT reduction, Abeta is known to induce morphological changes in astrocytes. Following addition of Abeta (20 microM), polygonal astrocytes changed into process-bearing stellate cells. To explore a possible linkage between these two effects of Abeta, we tested if astrocyte stellation is induced by agents that mimic the effect of Abeta on MTT reduction. Cholesterol (5 5000 nM) and lysophosphatidic acid (0.2-20 microg/ml) were found to accelerate the exocytosis of MTT formazan in a similar manner to Abeta, but failed to induce astrocyte stellation. Therefore, Abeta-induced inhibition of MTT reduction is unlikely to be directly linked to its effect on astrocyte morphology.

  10. Screen anticancer drug in vitro using resonance light scattering technique.

    PubMed

    Chen, Zhanguang; Liu, Guoliang; Chen, Meizhen; Xu, Benjie; Peng, Yurui; Chen, Maohuai; Wu, Mingyao

    2009-02-15

    An in vitro screening model using resonance light scattering (RLS) technique with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reagent as the reactive probe to target cancer cell was firstly developed. In this model, MTT was reduced by viable cancer cells to produce a purple formazan. Cell viability was proportional to the number of formazan induced strong light scattering signal. The inhibition rate of anticancer drug was found to vary inversely with the H(22)-MTT system RLS intensity. So it was intuitive to see the sequence of the tumor suppressive activity of six anticancer drugs without data processing by RLS/MTT screening spectra. Compared with the traditional MTT method, this method has high sensitivity, low detection limit and quite intuitive screening results which were identical to those obtained from the MTT colorimetric assay.

  11. Reduction of MTT to Purple Formazan by Vitamin E Isomers in the Absence of Cells.

    PubMed

    Lim, Su-Wen; Loh, Hwei-San; Ting, Kang-Nee; Bradshaw, Tracey Dawn; Allaudin, Zeenathul Nazariah

    2015-04-01

    The yellow tetrazolium salt 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) is widely used to determine cell viability in cell proliferation and cytotoxic assays. MTT is reduced by metabolically active cells to form an insoluble purple formazan product that is quantifiable by spectrophotometry. It is the most common and direct assay for cell viability. However, in this present study, we demonstrated that the vitamin E isomers α-β-γ-δ-tocotrienols and α-tocopherol were able to reduce MTT into a formazan product, despite the absence of living cells. For comparison, a second method for determining cell viability, which is the neutral red uptake assay, was used in parallel with the MTT assay. The results showed that neutral red did not interact with the vitamin E isomers. Our findings suggest that the MTT assay is not suitable for studying the proliferative effects of vitamin E isomers on cell growth.

  12. Reduction of MTT to Purple Formazan by Vitamin E Isomers in the Absence of Cells

    PubMed Central

    Lim, Su-Wen; Loh, Hwei-San; Ting, Kang-Nee; Bradshaw, Tracey Dawn; Allaudin, Zeenathul Nazariah

    2015-01-01

    The yellow tetrazolium salt 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) is widely used to determine cell viability in cell proliferation and cytotoxic assays. MTT is reduced by metabolically active cells to form an insoluble purple formazan product that is quantifiable by spectrophotometry. It is the most common and direct assay for cell viability. However, in this present study, we demonstrated that the vitamin E isomers α-β-γ-δ-tocotrienols and α-tocopherol were able to reduce MTT into a formazan product, despite the absence of living cells. For comparison, a second method for determining cell viability, which is the neutral red uptake assay, was used in parallel with the MTT assay. The results showed that neutral red did not interact with the vitamin E isomers. Our findings suggest that the MTT assay is not suitable for studying the proliferative effects of vitamin E isomers on cell growth. PMID:26868595

  13. Sensitivity of two in vitro assays for evaluating plant activity against the infective stage of Haemonchus contortus strains.

    PubMed

    Al-Rofaai, A; Rahman, W A; Abdulghani, Mahfoudh

    2013-02-01

    The sensitivity of larval paralysis assay (LPA) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide-formazan (MTT-formazan) assay was compared to evaluate the anthelmintic activity of plant extracts. In this study, the methanolic extract of Azadirachta indica (neem) was evaluated for its activity against the infective-stage larvae (L(3)) of susceptible and resistant Haemonchus contortus strains using the two aforementioned assays. In both in vitro assays, the same serial concentrations of the extract were used, and the median lethal concentrations were determined to compare the sensitivity of both assays. The results revealed a significant difference (P < 0.05) in the sensitivity of the LPA and the MTT-formazan assay. The MTT-formazan assay is more feasible for practical applications because it measured the L(3) mortality more accurately than LPA. This study may help find a suitable assay for investigating the anthelmintic activity of plant extracts against trichostrongylid nematodes.

  14. Sensitive detection of PDT-induced cell damages with luminescent oxygen nanosensors

    NASA Astrophysics Data System (ADS)

    Ma, Hong-Ru; Peng, Hong-shang; You, Fang-tian; Ping, Jian-tao; Zhou, Chao; Guo, Lan-ying

    2016-09-01

    In this work luminescent nanosensors specifically created for intracellular oxygen (ic-O2) were utilized to assess photodynamic therapy (PDT) -induced cell damages. Firstly, ic-O2 was demonstrated to be consumed much faster than extracellular O2 with respective O2 nanosensors. Using the ic-O2 nanosensors, PDT-treated cells with different degree of impairment were then resolved according to the oxygen consumption rate (OCR). The evolving trend of cytotoxicity derived from OCRs was in agreement with cell viability obtained from 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Moreover, the direct damage of PDT on cell mitochondria was successfully detected by monitoring respiration instantly after PDT treatment, which is actually beyond the scope of MTT assay. These results suggest that fluorescence sensing of ic-O2-associated cell respiration is promising and even may become a standardized method, complementary to MTT assay, to evaluate PDT-induced cytotoxicity.

  15. Vitamin A as an enzyme that catalyzes the reduction of MTT to formazan by vitamin C.

    PubMed

    Chakrabarti, R; Kundu, S; Kumar, S; Chakrabarti, R

    2000-09-18

    The tetrazolium salt 3(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) is reduced to formazan by the succinate dehydrogenase system of active mitochondria, and hence, specifically used to assay for the viable cells, such as measurement of cell proliferation, cytotoxicity, and cell number. However, in the present study we have shown that some component specifically present in M199 but not in RPMI 1640 media can reduce MTT to formazan in the absence of a living system. Further study revealed that ascorbic acid reduced MTT to formazan, which was profoundly increased by a very small amount of retinol, whereas retinol alone had no effect. Oxidation of ascorbic acid by H(2)O(2) destroyed its ability to reduce MTT. The rate of MTT reduction was directly proportional to the concentration of MTT in the absence of retinol, but approached a zero-order state beyond a certain concentration of MTT in the presence of retinol. Furthermore, retinol remained unchanged after the completion of the reaction. Taken together, these results showed that retinol acts as a reductase that catalyzes the reduction of MTT to formazan using ascorbic acid as the cosubstrate (electron donor).

  16. Cyanogen bromide

    Integrated Risk Information System (IRIS)

    Cyanogen bromide ; CASRN 506 - 68 - 3 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic

  17. Vinyl bromide

    Integrated Risk Information System (IRIS)

    Vinyl bromide ; CASRN 593 - 60 - 2 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic Eff

  18. An efficient and economical MTT assay for determining the antioxidant activity of plant natural product extracts and pure compounds.

    PubMed

    Liu, Yunbao; Nair, Muraleedharan G

    2010-07-23

    Antioxidants scavenge free radicals, singlet oxygen, and electrons in cellular redox reactions. The yellow MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide] is reduced to a purple formazan by mitochondrial enzymes. NADPH is the basis of established in vitro cell viability assays. An antioxidant assay has been developed utilizing the redox reaction between MTT and selected natural product extracts and purified compounds. This simple, fast, and inexpensive MTT antioxidant assay is comparable with the lipid peroxidation inhibitory assay and can be mechanized to achieve high throughput.

  19. Inhibitory and Cytotoxic Activities of Chrysin on Human Breast Adenocarcinoma Cells by Induction of Apoptosis

    PubMed Central

    Samarghandian, Saeed; Azimi-Nezhad, Mohsen; Borji, Abasalt; Hasanzadeh, Malihe; Jabbari, Farahzad; Farkhondeh, Tahereh; Samini, Mohammad

    2016-01-01

    Objectives: Chrysin, an active natural bioflavonoid found in honey and many plant extracts, was first known for its antioxidant and anti-inflammatory effects. The fact that antioxidants have several inhibitory effects against different diseases, such as cancer, led to search for food rich in antioxidants. In this study, we investigated the antiproliferative and apoptotic effects of chrysin on the cultured human breast cancer cells (MCF-7). Materials and Methods: Cells were cultured in Roswell Park Memorial Institute medium and treated with different chrysin concentrations for three consecutive days. Cell viability was quantitated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The percentage of apoptotic cells was determined by flow cytometry using Annexin V-fluorescein isothiocyanate. Results: The MTT assay showed that chrysin had an antiproliferative effect on MCF-7 cells in a dose- and time-dependent manner. The 50% cell growth inhibition values for chrysin against MCF-7 cells were 19.5 and 9.2 μM after 48 and 72 h, respectively. Chrysin induced apoptosis in MCF-7 cells as determined by flow cytometry. Chrysin inhibits the growth of the breast cancer cells by inducing cancer cell apoptosis which may, in part, explain its anticancer activity. Conclusion: This study shows that chrysin could also be considered as a promising chemotherapeutic agent and anticancer activity in treatment of the breast cancer cells in future. SUMMARY Chrysin had an antiproliferative effect on human breast cancer cells (MCF-7) cells in a dose- and time-dependent mannerChrysin induced apoptosis in MCF-7 cells, as determined by flow cytometryChrysin inhibits the growth of the breast cancer cells by inducing cancer cell apoptosisChrysin may have anticancer activity. Abbreviations used: Human breast cancer cells (MCF-7), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), phosphate-buffered saline (PBS), normal fibroblast mouse (L929). PMID

  20. In vitro cytotoxicity assays of solid lipid nanoparticles in epithelial and dermal cells

    NASA Astrophysics Data System (ADS)

    Ridolfi, D. M.; Marcato, P. D.; Machado, D.; Silva, R. A.; Justo, G. Z.; Durán, N.

    2011-07-01

    In recent years, the interest in nanostructured systems to drug delivery has increased because they offer several advantages over conventional dosage forms. Solid Lipid Nanoparticles (SLN) have been highlighted among these systems because they have advantages such as high physical stability, protection against drug degradation and ease of scale-up and manufacturing, without using organic solvent. The aim of this work was to evaluate the potential of SLN, by in vitro cytotoxicity assays, for dermal drug delivery. SLN of three different lipids were prepared by hot high pressure homogenization and the cytotoxicity was assessed by 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test in mouse 3T3 fibroblasts and human HaCaT keratinocytes. SLN showed no cytotoxic potential suggesting a great potential for dermal application.

  1. DEVD-Based Hydrogelator Minimizes Cellular Apoptosis Induction

    PubMed Central

    Tang, An-Ming; Wang, Wei-Juan; Mei, Bin; Hu, Wang-Lai; Wu, Mian; Liang, Gao-Lin

    2013-01-01

    Herein, we report the rational design of a DEVD-based heptapeptide hydrogelator 1 which is susceptible to caspase-3 (CASP3), and its isomeric control hydrogelator 2 with a DEDV-based heptapeptide sequence. Self-assembly of 1 in water results in flexuous, long nanofibers to form supramolecular hydrogel I with higher mechanical strength than that of hydrogel II which is composed of rigid, short nanofibers of 2. In vitro enzymatic analysis indicated that 1 is susceptive to CASP3 while 2 is not. 3-(4,5-dimethylthiazol-2-yl) 2,5 diphenyl tetrazolium bromide (MTT) and Western blot analyses indicated that DEDV-based hydrogelator 2 induces cell death via apoptotic pathway while the DEVD-based hydrogelator 1 minimizes cellular apoptosis induction. PMID:23673405

  2. Studies on Cytotoxic Activity against HepG-2 Cells of Naphthoquinones from Green Walnut Husks of Juglans mandshurica Maxim.

    PubMed

    Zhou, Yuanyuan; Yang, Bingyou; Jiang, Yanqiu; Liu, Zhaoxi; Liu, Yuxin; Wang, Xiaoli; Kuang, Haixue

    2015-08-26

    Twenty-seven naphthoquinones and their derivatives, including four new naphthalenyl glucosides and twenty-three known compounds, were isolated from green walnut husks, which came from Juglans mandshurica Maxim. The structures of four new naphthalenyl glucosides were elucidated based on extensive spectroscopic analyses. All of these compounds were evaluated for their cytotoxic activities against the growth of human cancer cells lines HepG-2 by MTT [3-(4,5-dimethylthiazo l-2-yl)-2,5 diphenyl tetrazolium bromide] assay. The results were shown that most naphthoquinones in an aglycone form exhibited better cytotoxicity in vitro than naphthalenyl glucosides with IC50 values in the range of 7.33-88.23 μM. Meanwhile, preliminary structure-activity relationships for these compounds were discussed.

  3. Molecular docking study, synthesis and biological evaluation of Schiff bases as Hsp90 inhibitors.

    PubMed

    Dutta Gupta, Sayan; Snigdha, D; Mazaira, Gisela I; Galigniana, Mario D; Subrahmanyam, C V S; Gowrishankar, N L; Raghavendra, N M

    2014-04-01

    Heat shock protein 90 (Hsp90) is an emerging attractive target for the discovery of novel cancer therapeutic agents. Docking methods are powerful in silico tools for lead generation and optimization. In our mission to rationally develop novel effective small molecules against Hsp90, we predicted the potency of our designed compounds by Sybyl surflex Geom X docking method. The results of the above studies revealed that Schiff bases derived from 2,4-dihydroxy benzaldehyde/5-chloro-2,4-dihydroxy benzaldehyde demonstrated effective binding with the protein. Subsequently, a few of them were synthesized (1-10) and characterized by IR, (1)HNMR and mass spectral analysis. The synthesized molecules were evaluated for their potential to suppress Hsp90 ATPase activity by Malachite green assay. The anticancer studies were performed by 3-(4,5-dimethythiazol- 2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay method. The software generated results was in satisfactory agreement with the evaluated biological activity.

  4. Synthesis and characterization of Bodipy functionalized magnetic iron oxide nanoparticles for potential bioimaging applications.

    PubMed

    Topel, Seda Demirel; Topel, Önder; Bostancıoğlu, R Beklem; Koparal, A Tansu

    2015-04-01

    Multifunctional magnetic nanoparticles were synthesized for potential bio-imaging applications. Uniform PEI coated magnetic Fe3O4 (PEI-Fe3O4) nanoparticles were prepared by a modified co-precipitation method and then covalently conjugated with a fluorophore molecule, Bodipy-5 by the DCC/DMAP coupling reaction. The covalent binding of Bodipy-5 to the PEI coated magnetic Fe3O4 nanoparticles were confirmed by means of FTIR and XPS measurements. The imaging ability of the Bodipy coated magnetic nanoparticles was determined on two human cancer cells, A549 (human lung adenocarcinoma epithelial) and Ishikawa (endometrial adenocarcinoma), for the first time. Cytotoxicity of BOD-MNPs was evaluated in both cancer cells and healthy human umbilical vein endothelial cell line (HUVEC) by standard MTT (3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. In vitro activities of the nanoparticles were also investigated.

  5. Synthesis of biocompatible glycodendrimer based on fluorescent perylene bisimides and its bioimaging.

    PubMed

    Wang, Ke-Rang; An, Hong-Wei; Rong, Rui-Xue; Cao, Zhi-Ran; Li, Xiao-Liu

    2014-04-01

    A novel water-soluble fluorescent glycodendrimer based on perylene bisimides is synthesized, which exhibits high fluorescence quantum yield of 54%. While the binding interactions of PBI-Man with Concanavalin A (Con A) are studied by fluorescence spectra and CD spectra, which show strong binding affinity for Con A with the binding constant of 3.8 × 10(7) m(-1) for monomeric mannose, nearly four orders of magnitude higher affinity than the monovalent mannose ligand. Furthermore, the fluorescence imaging of macrophage cell with PBI-Man is investigated, and shows selectively binding interaction with the mannose receptor-medicated cell entry. Moreover, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) activities of PBI-Man show that PBI-Man as a biocompatible agent is noncytotoxic to living cells.

  6. Antitumor and antioxidant activity of protocatechualdehyde produced from Streptomyces lincolnensis M-20.

    PubMed

    Kim, Kyoung-Ja; Kim, Mi-Ae; Jung, Jee-Hyung

    2008-12-01

    We characterized the biological functions of protocatechualdehyde (PA) isolated from the butanol extract of culture supernatant from Streptomyces lincolnensis M-20. Following butanol extraction, it was purified by silica gel and Sephadex LH-20 column chromatography. PA was analyzed by Furier Transform Infrared Spectroscopy (FT-IR), Gas chromatograph-Mass Spectrometer (GC-MS), and Nuclear Magnetic Resonance (NMR). PA had potent antioxidant activity, as measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. Antitumor activity against MCF-7 human breast cancer cells was evaluated by the 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium-bromide (MTT) assay. PA treatment (0 approximately 150 muM) dose-dependently blocked apoptosis, as shown by improved cell viability and inter-nucleosomal DNA fragmentation. Our findings suggest that Streptomyces lincolnensis M-20, a lincomycin producer, also produces protocatechualdehyde.

  7. Exploring the dark side of MTT viability assay of cells cultured onto electrospun PLGA-based composite nanofibrous scaffolding materials.

    PubMed

    Qi, Ruiling; Shen, Mingwu; Cao, Xueyan; Guo, Rui; Tian, Xuejiao; Yu, Jianyong; Shi, Xiangyang

    2011-07-21

    One major method used to evaluate the biocompatibility of porous tissue engineering scaffolding materials is MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The MTT cell viability assay is based on the absorbance of the dissolved MTT formazan crystals formed in living cells, which is proportional to the number of viable cells. Due to the strong dye sorption capability of porous scaffolding materials, we propose that the cell viability determined from the MTT assay is likely to give a false negative result. In this study, we aim to explore the effect of the adsorption of MTT formazan on the accuracy of the viability assay of cells cultured onto porous electrospun poly(lactic-co-glycolic acid) (PLGA) nanofibers, HNTs (halloysite nanotubes)/PLGA, and CNTs (multiwalled carbon nanotubes)/PLGA composite nanofibrous mats. The morphology of electrospun nanofibers and L929 mouse fibroblasts cultured onto the nanofibrous scaffolds were observed using scanning electron microscopy. The viability of cells proliferated for 3 days was evaluated through the MTT assay. In the meantime, the adsorption of MTT formazan onto the same electrospun nanofibers was evaluated and the standard concentration-absorbance curve was obtained in order to quantify the contribution of the adsorbed MTT formazan during the MTT cell viability assay. We show that the PLGA, and the HNTs- or CNTs-doped PLGA nanofibers display appreciable MTT formazan dye sorption, corresponding to 35.6-50.2% deviation from the real cell viability assay data. The better dye sorption capability of the nanofibers leads to further deviation from the real cell viability. Our study gives a general insight into accurate MTT cytotoxicity assessment of various porous tissue engineering scaffolding materials, and may be applicable to other colorimetric assays for analyzing the biological properties of porous scaffolding materials.

  8. Evaluation of Direct Colorimetric MTT Assay for Rapid Detection of Rifampicin and Isoniazid Resistance in Mycobacterium tuberculosis

    PubMed Central

    Woldemeskel, Dawit; Gessesse, Amare

    2016-01-01

    With the spread of multidrug-resistant tuberculosis (MDR-TB) strains there is an increasing need for new accurate and cost-effective methods for a rapid diagnostic and drug susceptibility testing (DST), particularly in low-income countries where tuberculosis is hyperendemic. A colorimetric assay using 3-(4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide (MTT) has been suggested as a promising method for DST, especially to rifampicin. In this study, we standardized and evaluated the MTT assay for a rapid direct detection of rifampicin and isoniazid resistant Mycobacterium tuberculosis strains from sputum specimens using Lowenstein-Jensen (LJ) culture medium as a gold standard. The MTT assay sensitivity, specificity, positive and negative predictive values for rifampicin were 100%, 86%, 100%, 99%, respectively. For isoniazid, the MTT assay had a 100% sensitivity, specificity, positive and negative predictive values. Interestingly, the MTT assay gave interpretable results within two weeks for 94% of the samples compared to 7–14 weeks for LJ media. Overall, an excellent agreement was observed between MTT assay and LJ proportion method (Kappa, 0.91 for rifampicin and 1.00 for isoniazid). In conclusion, the direct colorimetric MTT assay simultaneously detects susceptible and resistant strains of M. tuberculosis within three weeks. It significantly shortens the time required to obtain a DST result and could be a reliable alternative method for rapid detection of drug-resistant TB strains in high-TB-burden resource-limited settings. PMID:28030634

  9. Ipratropium bromide HFA.

    PubMed

    Wellington, Keri

    2005-01-01

    Ipratropium bromide is a nonselective antagonist of the muscarinic receptors located on airway smooth muscle, and is delivered via a metered-dose inhaler (MDI). Because of the requirement to phase out chlorofluorocarbon (CFC)-propelled MDIs, the ipratropium bromide inhalation aerosol MDI has been redesigned with a hydrofluoroalkane as the propellant (ipratropium bromide HFA). Ipratropium bromide HFA has recently been approved in the US for the maintenance treatment of bronchospasm associated with COPD. Ipratropium bromide HFA 42 microg four times daily (one dose [42 microg] is delivered via two puffs of the inhaler) demonstrated comparable efficacy to that of ipratropium bromide CFC 42 microg four times daily, as measured by spirometric testing, in a large, randomized, double-blind, placebo-controlled, 12-week trial in patients with stable COPD. Similarly, four-times-daily ipratropium bromide HFA 42 microg and ipratropium bromide CFC 42 microg provided a comparable degree of bronchodilation in patients with stable COPD during a 1-year, open-label study primarily designed to assess safety. In both studies, the tolerability profiles of ipratropium bromide HFA and ipratropium bromide CFC were comparable. The most common adverse events were related to respiratory system disorders. During the 1-year study, dry mouth was reported by 1.3% and 0.7% of patients in the ipratropium bromide HFA or ipratropium bromide CFC groups.

  10. Relationship of MTT reduction to stimulants of muscle metabolism.

    PubMed

    Newman, J M; DiMaria, C A; Rattigan, S; Steen, J T; Miller, K A; Eldershaw, T P; Clark, M G

    2000-10-16

    MTT, a positively charged tetrazolium salt, is widely used as an indicator of cell viability and metabolism and has potential for histochemical identification of tissue regions of hypermetabolism. In the present study, MTT was infused in the constant-flow perfused rat hindlimb to assess the effect of various agents and particularly vasoconstrictors that increase muscle metabolism. Reduction of MTT to the insoluble formazan in muscles assessed at the end of experiments was linear over a 30 min period and production rates were greater in red fibre types than white fibre types. The vasoconstrictors, norepinephrine (100 nM) and angiotensin (10 nM) decreased MTT formazan production in all muscles but increased hindlimb oxygen uptake and lactate efflux. Veratridine, a Na(+) channel opener that increases hindlimb oxygen uptake and lactate efflux without increases in perfusion pressure, also decreased MTT formazan production. Membrane stabilizing doses (100 microM) of (+/-)-propranolol reversed the inhibitory effects of angiotensin and veratridine on MTT formazan production. Muscle contractions elicited by stimulation of the sciatic nerve, reversed the norepinephrine-mediated inhibitory effects on MTT formazan production, even though oxygen consumption and lactate efflux were further stimulated. Stimulation of hindlimb muscle oxygen uptake by pentachlorophenol, a mitochondrial uncoupler, was not associated with alterations in MTT formazan production. It is concluded that apart from muscle contractions MTT formazan production does not increase with increased muscle metabolism. Since the vasoconstrictors angiotensin and norepinephrine as well as veratridine activate Na(+) channels and the Na(+)/K(+) pump, energy required for Na(+) pumping may be required for MTT reduction. It is unlikely that vasoconstrictors that stimulate oxygen uptake do so by uncoupling respiration.

  11. The noncellular reduction of MTT tetrazolium salt by TiO₂ nanoparticles and its implications for cytotoxicity assays.

    PubMed

    Lupu, A R; Popescu, T

    2013-08-01

    We report results of noncellular tests, revealing the occurrence of photocatalytic interactions between titanium dioxide (TiO2, titania) nanoparticles and the MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide] cytotoxicity indicator. These interactions induce the reduction of MTT and formation of purple formazan under biologically relevant conditions. Classical MTT assays have been performed to evaluate the production of formazan in DMEM-F12 and RPMI-1640 cell culture media (containing 10% fetal bovine serum-FBS) treated with Degussa-P25 TiO2 nanoparticles, in the absence of cells. The colorimetric determinations revealed the noncellular MTT to formazan transformation induced by TiO2 nanoparticles, under conditions commonly used for in vitro cytotoxicity testing of nanomaterials. The formazan precipitation was found to be proportional to the TiO2 concentration, being enhanced under laboratory daylight exposure. The photocatalytic nature of the studied effect was assessed under UV irradiation at 365nm. The biological significance of the reported reaction was established with respect to cellular reference experiments performed on V79-4, HeLa and B16 cell lines. The results show false viability increases with up to 14% (for TiO2 concentrations generally higher than 50μg/ml), induced by the TiO2-MTT reaction. This type of artifacts may lead to underestimated toxicity or false proliferation results.

  12. Backscattered light confocal imaging of intracellular MTT-formazan crystals.

    PubMed

    Bernas, Tytus; Dobrucki, Jurek W

    2004-06-01

    Metabolically active animal and plant cells reduce MTT tetrazolium salt to a corresponding nonfluorescent formazan. Reduction of MTT by viable cells is exploited in a number of tests widely used in biological research. The aim of this study was to optimize a microscopy method of detecting small crystals of MTT-formazan formed in intact cells maintained in in vitro cultures. We examined scattering properties of small intracellular crystals of MTT formazan and found that the efficiency of light scattering was dependent on wavelength. Small (<3 microm) crystals of MTT-formazan, formed in viable cells, scattered red, but not blue, light. Large crystals, which are formed later at a stage when cells begin to lose viability, scattered both red and blue light. We conclude that optimal detection of early stages of crystallization of MTT-formazan in living cells is possible using confocal microscopy of red, but not blue, scattered light. High contrast and resolution of images can be achieved by filtering out interference effects in the frequency domain.

  13. Serum albumin leads to false-positive results in the XTT and the MTT assay.

    PubMed

    Funk, Dorothee; Schrenk, Hans-Hermann; Frei, Eva

    2007-08-01

    Tetrazolium salts like 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) or sodium 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) that form formazans after reduction are widely used to investigate cell viability. Besides cellular enzymes, some constituents of cell media and other substances reduce tetrazolium salts, thereby interfering with these assays. We describe here that different preparations of serum albumin from bovine or human origin can lead to a concentration-dependent increase in the signals of the XTT assay; therefore leading to an overestimation of cell numbers and to an underestimation of potential cytotoxic effects of compounds to be tested. The same effect was seen in the MTT assay with human serum albumin (HSA). We demonstrate that this reductive activity cannot be inactivated by proteolytic digestion, but that it is due to the free cysteine residue in albumin, and is also observed when cysteine or glutathione (GSH) are used. Binding of N-ethylmaleimide (NEM) to the free cysteine residue leads to a decrease of the albumin interference in the XTT assay.

  14. Colorimetric method for rapid detection of Oxacillin resistance in Staphylococcus aureus and its comparison with PCR for mec A gene

    PubMed Central

    Ghanwate, Niraj; Thakare, Prashant; Bhise, P. R.; Gawande, Sonali

    2016-01-01

    Rapid and accurate detection of Methicillin Resistant Staphylococcus aureus (MRSA) is an important role of clinical microbiology laboratories to avoid treatment failure. The detection of MRSA is based on phenotypic assays which require at least 24 h to perform. Detection of the mecA gene or of PBP 2a is the “gold standard”, but not always available. The aim of this study was to evaluate a rapid method for detection of MRSA by using 3 (4, 5 dimethyl thiazole -2-yl) -2, 5 diphenyl tetrazolium bromide (MTT). Total 126 isolates of MRSA were collected from tertiary healthcare center and were confirmed by oxacillin screening agar test as per CLSI guidelines. Amplification of mecA gene was performed by using PCR. MTT assay was carried out for all the isolates in 96 well Microtitre plate and compared with standard methods of CLSI. Out of 126 isolates, 98 were found to be mecA positive. MTT method was found to be 98.98% sensitive and 96.43% specific. The MTT based colorimetric method is rapid and simple test for screening of oxacillin resistance in Staphylococcus aureus. It significantly shortens the time to just 7 h required to obtained a drug susceptibility test and could be useful to screen MRSA. PMID:26960268

  15. Localization of MTT formazan in lipid droplets. An alternative hypothesis about the nature of formazan granules and aggregates.

    PubMed

    Diaz, Giacomo; Melis, M; Musin, A; Piludu, M; Piras, M; Falchi, A M

    2007-01-01

    MTT (3-(4, 5-dimethyl-2-thiazolyl)-2, 5-dihphenyltetrazolium bromide) assay is a widely used method to assess cell viability and proliferation. MTT is readily taken up by cells and enzymatically reduced to formazan, a dark compound which accumulates in cytoplasmic granules. Formazan is later eliminated by the cell by a mechanisms often indicated as exocytosis, that produces characteristic needle-like aggregates on the cell surface. The shape of formazan aggregates and the rate of exocytosis change in the presence of bioactive amyloid beta peptides (Abeta) and cholesterol. Though the cellular mechanisms involved in MTT reduction have been extensively investigated, the exact nature of formazan granules and the process of exocytosis are still obscure. Using Nile Red, which stains differentially neutral and polar lipids, and a fluorescent analog of cholesterol (NBD-cholesterol), we found that formazan localized in lipid droplets, consistent with the lipophilic nature of formazan. However, formazan granules and aggregates were also found to form after killing cells with paraformaldehyde fixation. Moreover, formazan aggregates were also obtained in cell-free media, using ascorbic acid to reduce MTT. The density and shape of formazan aggregates obtained in cell-free media was sensitive to cholesterol and Abeta. In cells, electron microscopy failed to detect the presence of secretory vesicles, but revealed unusual fibers of 50 nm of diameter extending throughout the cytoplasm. Taken together, these findings suggest that formazan efflux is driven by physico-chemical interactions at molecular level without involving higher cytological mechanisms.

  16. Evidence of ATP assay as an appropriate alternative of MTT assay for cytotoxicity of secondary effluents from WWTPs.

    PubMed

    Yang, Yang; Lu, Yun; Wu, Qian-Yuan; Hu, Hong-Ying; Chen, Ying-Hua; Liu, Wan-Li

    2015-12-01

    Biological tests are effective and comprehensive methods to assess toxicity of environmental pollutants to ensure the safety of reclaimed water. In this study, the canonical MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was performed to evaluate the cytotoxicity of dissolved organic matters (DOMs) of secondary effluents from wastewater treatment plants (WWTPs). It was surprising that most concentrated DOMs treated HepG2 cells yielded much higher signal compared with vehicle control regardless of difference of treatment technologies and seasons. However, there was actually no obvious enhancement of the cell proliferation by microscopy. In order to find out potential reason for the discrepancy, another three assays were performed. The results of ATP assay and flow cytometry showed expected toxicity, which was consistent with microscopy and previous studies, while DNA assay did not exhibit apparent change in treated cells. The possible mechanisms of abnormal MTT signal could be that some materials in secondary effluents isolated by solid extraction with HLB resin directly reacted with MTT and/or enhanced the activity of mitochondrial dehydrogenase. Therefore, the MTT assay is not suitable to assess cytotoxicity of complex mixtures such as secondary effluents, while ATP assay is an optional sensitive method. This study also suggests the importance of choosing both suitable extraction methods and detection assays for toxicity evaluation of component-unknown environmental samples.

  17. Microscopic analysis of MTT stained boar sperm cells.

    PubMed

    van den Berg, B M

    2015-01-01

    The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few min in a diffuse staining of the midpiece of the sperm cells. Upon further incubation the staining of the midpiece became more intense, and gradually the formation of packed formazan granules became more visible. At the same time, a small formazan stained granule appeared medially on the sperm head, which increased in size during further incubation. After incubation for about 1 h the midpiece granules were intensely stained and more clearly distinct as granules, while aggregation of sperm cells occurred. Around 90% of the sperm cells showed these staining events. At the end of the staining the formazan granules have disappeared from both the sperm cells and medium, whereas formazan crystals appeared as thin crystal threads, that became heavily aggregated in the incubation medium. It was concluded that formazan is taken up by lipid droplets in the cytoplasm. Further, the use of the MTT assay to test for sperm viability should be regarded as a qualitative assay, whereas its practical use at artificial insemination (AI) Stations is limited.

  18. Inhibition of Pro-inflammatory Mediators and Cytokines by Chlorella Vulgaris Extracts

    PubMed Central

    Sibi, G.; Rabina, Santa

    2016-01-01

    Objective: The aim of this study was to determine the in vitro anti-inflammatory activities of solvent fractions from Chlorella vulgaris by inhibiting the production of pro-inflammatory mediators and cytokines. Methods: Methanolic extracts (80%) of C. vulgaris were prepared and partitioned with solvents of increasing polarity viz., n-hexane, chloroform, ethanol, and water. Various concentrations of the fractions were tested for cytotoxicity in RAW 264.7 cells using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and the concentrations inducing cell growth inhibition by about 50% (IC50) were chosen for further studies. Lipopolysaccharide (LPS) stimulated RAW 264.7 cells were treated with varying concentrations of C. vulgaris fractions and examined for its effects on nitric oxide (NO) production by Griess assay. The release of prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin 6 (IL-6) were quantified using enzyme-linked immunosorbent assay using Celecoxib and polymyxin B as positive controls. Results: MTT assay revealed all the solvent fractions that inhibited cell growth in a dose-dependent manner. Of all the extracts, 80% methanolic extract exhibited the strongest anti-inflammatory activity by inhibiting NO production (P < 0.01), PGE2 (P < 0.05), TNF-α, and IL-6 (P < 0.001) release in LPS induced RAW 264.7 cells. Both hexane and chloroform fractions recorded a significant (P < 0.05) and dose-dependent inhibition of LPS induced inflammatory mediators and cytokines in vitro. The anti-inflammatory effect of ethanol and aqueous extracts was not significant in the study. Conclusion: The significant inhibition of inflammatory mediators and cytokines by fractions from C. vulgaris suggests that this microalga would be a potential source of developing anti-inflammatory agents and a good alternate for conventional steroidal and nonsteroidal anti-inflammatory drugs. SUMMARY C. vulgaris extracts have potential anti

  19. A rapid colorimetric assay for the quantitation of the viability of free-living larvae of nematodes in vitro.

    PubMed

    James, Catherine E; Davey, Mary W

    2007-09-01

    With increasing drug resistance in gastrointestinal parasites, identification of new anthelmintics is essential. The non-parasitic nematode Caenorhabditis elegans is used extensively as a model to identify drug targets and potential novel anthelmintics because it can be readily cultured in vitro. Traditionally, the assessment of worm viability has relied on labour-intensive developmental and behavioral assays. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide-formazan (MTT-formazan) colorimetric assay uses metabolic activity as a marker of viability in mammalian cell culture systems and has been applied for use with filarial nematodes. In the present study, this assay has been optimized and validated to rapidly assess the viability of C. elegans after drug treatment. Living, but not dead, C. elegans take up MTT and reduce it to the blue formazan, providing visual, qualitative, and quantitative assessment of viability. MTT at a concentration of 5 mg/ml with 3 h incubation was optimal for detecting changes in viability with drug treatment. We have applied this assay to quantitate the effects of ivermectin and short-chain alcohols on the viability of C. elegans. This assay is also applicable to first-stage larvae of the parasitic nematode Haemonchus contortus. The advantage of this assay is the rapid quantitation in screening drugs to identify potential anthelmintics.

  20. Mercury Bromide Laser Research.

    DTIC Science & Technology

    1981-05-04

    Discharge", Optics Lett., 2(3), (March 1978). 7. R. Burnham, "Discharge Pumped Mercuric Halide Dissociation Lasers", Appl. Phys. Lett., 33: 15 (July 1978...laser and fluorescence signals. Neutral density filters served to prevent saturation of the detector during the laser measurements. F. Laser Output as a...REFERENCES . E. J. Schimitschek and J. E. Celto, " Mercuric Bromide Dissociation Laser in an Electric Discharge," Optics Lett. 2(3), March 1978. This

  1. Electrochemical sensors, MTT and immunofluorescence assays for monitoring the proliferation effects of cissus populnea extracts on Sertoli cells

    PubMed Central

    2011-01-01

    Background We describe the development of an electrochemical sensor array for monitoring the proliferation effects of cissus populnea plant extracts on TM4 Sertoli cells. Methods The proliferation activities of the extracts on Sertoli cells were studied using a high-throughput electrochemical sensor array (DOX-96) and the analytical sensor characteristics were compared with conventional colorimetric MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and fluorescence spectroscopy. Results This work shows that there is a definite positive trend in the proliferation effect of the extract of Cissus populnea on the TM4 Sertoli cells. All of the three techniques confirmed that the most effective concentration for the proliferation is 10 ppm. At this concentration, the proliferation effect was established around 120% for both DOX-96 and MTT techniques, whereas fluorescence assays showed a higher level (120-150%). DOX-96 showed a lower limit of detection (1.25 × 10(4) cells/ml); whereas the LOD recorded for both MTT and fluorescence techniques was 2.5 × 10(4) cells/ml. Visual examination of the cells by means of confocal fluorescence microscopy confirmed the proliferation of Sertoli cells as was determined using the MTT assay. This investigation provides a confident interpretation of the results and proved that the most effective concentration for the proliferation using Cissus populnea plant extract is 10 ppm. Conclusions Overall, the DOX results compared well with the conventional methods of checking proliferation of cells. The fascinating feature of the sensor array is the ability to provide continuous proliferation experiments with no additional reagents including 96 simultaneous electrochemical experiments. The use of the DOX-96 could reduce a typical bioassay time by 20-fold. Thus the DOX-96 can be used as both a research tool and for practical cell culture monitoring. PMID:21575213

  2. Antiamnesic effects of ethyl acetate fraction from chestnut (Castanea crenata var. dulcis) inner skin on Aβ(25-35)-induced cognitive deficits in mice.

    PubMed

    Jeong, Hee-Rok; Jo, Yu Na; Jeong, Ji Hee; Jin, Dong Eun; Song, Byung Gi; Choi, Soo Jung; Shin, Dong-Hoon; Heo, Ho Jin

    2012-12-01

    To investigate neuronal cell protective effects of an ethyl acetate fraction from chestnut inner skin, in vitro assays, including 2',7'-dichlorofluorescein diacetate, 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT), and lactate dehydrogenase (LDH), were performed. Intracellular accumulation of reactive oxygen species resulting from hydrogen peroxide (H(2)O(2)) treatment of PC12 cells was significantly reduced when ethyl acetate fractions were present in the medium compared to PC12 cells treated with H(2)O(2) only. In a cell viability assay using MTT, the ethyl acetate fraction protected against H(2)O(2)-induced neurotoxicity, and inhibited LDH release into the medium. In addition, the ethyl acetate fraction improved in vivo cognitive ability against amyloid β-peptide (Aβ)-induced neuronal deficit. High-performance liquid chromatography analyses showed that gallic acid, catechin, and epicatechin were predominant phenolics in the ethyl acetate fraction. Consequently, the results suggest that chestnut inner skin, including above phenolics, could ameliorate Aβ-induced learning and memory deficiency, and be utilized as effective substances for neurodegenerative disorders, notably Alzheimer's disease.

  3. Multinucleated Giant Cancer Cells Produced in Response to Ionizing Radiation Retain Viability and Replicate Their Genome

    PubMed Central

    Mirzayans, Razmik; Andrais, Bonnie; Scott, April; Wang, Ying W.; Kumar, Piyush; Murray, David

    2017-01-01

    Loss of wild-type p53 function is widely accepted to be permissive for the development of multinucleated giant cells. However, whether therapy-induced multinucleation is associated with cancer cell death or survival remains controversial. Herein, we demonstrate that exposure of p53-deficient or p21WAF1 (p21)-deficient solid tumor-derived cell lines to ionizing radiation (between 2 and 8 Gy) results in the development of multinucleated giant cells that remain adherent to the culture dish for long times post-irradiation. Somewhat surprisingly, single-cell observations revealed that virtually all multinucleated giant cells that remain adherent for the duration of the experiments (up to three weeks post-irradiation) retain viability and metabolize 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), and the majority (>60%) exhibit DNA synthesis. We further report that treatment of multinucleated giant cells with pharmacological activators of apoptosis (e.g., sodium salicylate) triggers their demise. Our observations reinforce the notion that radiation-induced multinucleation may reflect a survival mechanism for p53/p21-deficient cancer cells. With respect to evaluating radiosensitivity, our observations underscore the importance of single-cell experimental approaches (e.g., single-cell MTT) as the creation of viable multinucleated giant cells complicates the interpretation of the experimental data obtained by commonly-used multi-well plate colorimetric assays. PMID:28208747

  4. Differential protection of pre-, co- and post-treatment of curcumin against hydrogen peroxide in PC12 cells.

    PubMed

    Siddiqui, M A; Kashyap, M P; Kumar, V; Tripathi, V K; Khanna, V K; Yadav, S; Pant, A B

    2011-03-01

    Pharmacological potential of curcumin was assessed in PC12 cells against hydrogen peroxide (H(2) O(2)) exposure. In MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and lactate dehydrogenase (LDH) assays, 24-hour exposure of H(2)O(2) (0.5 mM and above) was found to be cytotoxic. A significant (p < 0.001) increase in percentage cell viability was recorded in PC12 cells pretreated with curcumin (25, 50 and 100 µg/mL) for 24 hours prior to H(2)O(2) (0.5 and 1 mM) exposure for 24 hours. Co-exposure to H(2)O(2) and curcumin was also found effective. However, a therapeutic treatment of curcumin for 24 hours after H(2)O(2) exposure to the cells was found ineffective. Differential response of PC12-H(2)O(2) model to curcumin in MTT and LDH assays suggests the utility of these endpoints to sort the drug candidates to study their antioxidant potential.

  5. Decrease in toxic potential of mixed tensides maintained below the critical micelle concentration: an in vitro study.

    PubMed

    Benassi, L; Bertazzoni, G; Magnoni, C; Rinaldi, M; Fontanesi, C; Seidenari, S

    2003-01-01

    Sodium lauryl sulphate (SLS) is an anionic tenside widely utilized in commercial topical preparations that may cause skin irritation. It has been shown that the barrier damage caused by SLS in vivo is lower when SLS is used in combination with other tensides which are able to reduce the critical micelle concentration (CMC). The aim of our study was to evaluate if the cytotoxic effect of SLS is reduced by the association with different tensides also at concentrations well below the CMC. Normal human keratinocytes from plastic surgery were grown in serum-free medium. At subconfluency, the cells were treated with SLS at a dose of 0.0025% in combination with cocamidopropyl betaine, Tween 20 and Tween 80 at the minimum toxic dose. Following tenside treatment, the culture medium was changed, and after 24 h the cells were collected for (3)H-thymidine incorporation, the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay and neutral red (NR) uptake. The cytotoxic effect on normal human keratinocytes, as evaluated by (3)H-thymidine incorporation, MTT assay and NR uptake, was significantly decreased by the combination with all the tested tensides. The correlation between cytotoxicity and physical properties was also studied by a conductimetric assay to investigate the mechanism involved in this toxicity reduction.

  6. Detrimental effect of fast neutrons on cultured immature rat hippocampal cells: relative biological effectiveness of in vitro cell death indices.

    PubMed

    Yang, M; Kim, J S; Son, Y; Kim, J; Kim, J Y; Kim, S H; Kim, J C; Shin, T; Moon, C

    2011-09-01

    This in vitro study compared the detrimental effect and relative biological effectiveness (RBE) of high-linear energy transfer (LET) fast neutrons on rat immature hippocampal cultured cells with those of low-LET γ rays. Immature hippocampal cells were exposed to fast neutrons or γ rays. Cytotoxicity and cell viability were analyzed using a lactate dehydrogenase (LDH)-release assay and a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay, respectively. The cytotoxicity and cell viability with fast neutrons or γ rays varied in a dose-dependent pattern. In the LDH release and MTT assay indices, the RBEs of fast neutrons were approximately 2.35 and 2.42, respectively. Fast neutrons markedly induced apoptotic changes in immature hippocampal cells with increased expression of active caspase-3 and cleaved poly(ADP-ribose) polymerase. Increased cytotoxicity and decreased cell viability in immature hippocampal cells were seen in a dose-dependent pattern after fast-neutron and γ irradiation. Fast neutrons have a higher RBE for cell death indices than γ rays.

  7. Allium sativum L. extract prevents methyl mercury-induced cytotoxicity in peripheral blood leukocytes (LS).

    PubMed

    Abdalla, F H; Bellé, L P; De Bona, K S; Bitencourt, P E R; Pigatto, A S; Moretto, M B

    2010-01-01

    Adenosine deaminase (ADA) is involved in purine metabolism and plays a significant role in the immune system. The focus of this investigation was to examine the effects of low concentrations of organic mercury on ADA activity in human leukocytes and to investigate the relationship between these effects and cell death. We have examined the protective potential effects of Allium sativum extract (GaE) against Methylmercury (MeHg)-induced cytotoxic effects on human leucocytes under in vitro conditions. MeHg (0.05-10 microM) significantly decreased leukocyte viability (58.97% for MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) and 51.67% for Alamar Blue (AB) and this decrease was positively correlated to the MeHg-induced inhibition of ADA activity. N-acetylcysteine (NAC) and GaE prevented both the MeHg-induced cytotoxic effects on leukocytes according to MTT and AB assays and the effects on the ADA activity. The present results suggest that the protective effects of GaE against MeHg-induced leukocyte damage is related to the removal of oxidant species generated in the presence of MeHg due to the antioxidant efficacy of garlic constituents. It is important to point out that the intense presence of ADA in Leukocyte suspension (LS) highlights the relevant effects in the immune system and in vitro cytotoxicity of MeHg exposure.

  8. A p-menth-1-ene-4,7-diol (EC-1) from Eucalyptus camaldulensis Dhnh. triggers apoptosis and cell cycle changes in Ehrlich ascites carcinoma cells.

    PubMed

    Islam, Farhadul; Khanam, Jahan Ara; Khatun, Mahbuba; Zuberi, Natasha; Khatun, Laboni; Kabir, Syed Rashel; Reza, Md Abu; Ali, M M; Rabbi, M A; Gopalan, Vinod; Lam, Alfred King-Yin

    2015-04-01

    Anticancer activities of p-menth-1-ene-4,7-diol (EC-1) isolated from Eucalyptus camaldulensis Dhnh. were studied on Ehrlich ascites carcinoma (EAC) cells by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. Anticancer activities also analyzed in EAC-bearing mice by assessment of cancer growth inhibition, changes in cancer volume, changes in life span, and hematological parameters. Apoptosis was analyzed by fluorescence microscope, DNA fragmentation assay, and flow cytometry. The expression of apoptosis-related genes, Bcl-2, Bcl-X, PARP-1, p53, and Bax, were analyzed using polymerase chain reaction (PCR). EC-1 significantly inhibited proliferation of EAC cells in vivo and restored the altered hematological parameters of EAC-bearing mice. Cytological observation by fluorescence microscope showed apoptosis of EAC cells upon treatment with EC-1. Also, DNA fragmentation assay revealed EAC cells' apoptosis following EC-1 treatment. Increased mRNA expressions of p53 and Bax genes and negative expressions of Bcl-2 and Bcl-X were observed in cells treated with EC-1. These findings confirmed the induction of apoptosis by EC-1. In addition, MTT assay showed dose-dependent anticancer activity of EC-1 against EAC cell. Cell cycle analysis revealed that EC-1 treatment caused suppression of EAC cells at S phase. To conclude, EC-1 is a novel anticancer compound and showed antiproliferative and apoptotic activities in cellular and mice models.

  9. Managing Nematodes without Methyl Bromide

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methyl bromide is an effective pre-plant soil fumigant used to control nematodes in many high-input, high-value production systems including vegetables, nurseries, ornamentals, tree fruits, strawberries, and grapes. Because methyl bromide has provided a reliable return on investment for nematode c...

  10. Iron bromide vapor laser

    NASA Astrophysics Data System (ADS)

    Sukhanov, V. B.; Shiyanov, D. V.; Trigub, M. V.; Dimaki, V. A.; Evtushenko, G. S.

    2016-03-01

    We have studied the characteristics of a pulsed gas-discharge laser on iron bromide vapor generating radiation with a wavelength of 452.9 nm at a pulse repetition frequency (PRF) of 5-30 kHz. The maximum output power amounted to 10 mW at a PRF within 5-15 kHz for a voltage of 20-25 kV applied to electrodes of the discharge tube. Addition of HBr to the medium produced leveling of the radial profile of emission. Initial weak lasing at a wavelength of 868.9 nm was observed for the first time, which ceased with buildup of the main 452.9-nm line.

  11. Anticancer Activity of Chloroform Extract and Sub-fractions of Nepeta deflersiana on Human Breast and Lung Cancer Cells: An In vitro Cytotoxicity Assessment

    PubMed Central

    Al-Oqail, Mai M.; Al-Sheddi, Ebtesam S.; Siddiqui, Maqsood A.; Musarrat, Javed; Al-Khedhairy, Abdulaziz A.; Farshori, Nida N.

    2015-01-01

    relatively higher cytotoxicity. The A-549 cells were found to be more sensitive as compared to the MCF-7 cells. Abbreviations used: MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; NRU: Neutral red uptake; DMEM: Dulbecco's modified eagle medium; FBS: Fetal bovine serum; PBS: Phosphate buffer saline; DMSO: Dimethyl sulfoxide PMID:27013801

  12. The primary role of iron-mediated lipid peroxidation in the differential cytotoxicity caused by two varieties of talc nanoparticles on A549 cells and lipid peroxidation inhibitory effect exerted by ascorbic acid.

    PubMed

    Akhtar, Mohd Javed; Kumar, Sudhir; Murthy, Ramesh Chandra; Ashquin, Mohd; Khan, Mohd Imran; Patil, Govil; Ahmad, Iqbal

    2010-06-01

    Talc particles, the basic ingredient in different kinds of talc-based cosmetic and pharmaceutical products, pose a health risk to pulmonary and ovarian systems due to domestic and occupational exposures. Two types of talc nanoparticles depending on the source of geographical origin - indigenous- and commercial talc nanoparticles were assessed for their potential in vitro toxicity on A(549) cells; along with indigenous conventionally used microtalc particles. Cell viability, determined through live/dead staining and 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, decreased as a function of concentration, origin and size of particles. Both varieties of talc nanoparticles differentially induced lipid peroxidation (LPO), which was correlated with the pattern of lactate dehydrogenase (LDH) leakage, reactive oxygen species (ROS) generation, and glutathione (GSH) depletion. Relatively higher cytotoxicity of indigenous nanotalc could be attributed to its higher content of iron as compared to commercial nanotalc. The known scavenger of ROS, l-ascorbic acid significantly inhibited LPO induction due to talc particles. Data suggest that nanotalc toxicity on A(549) cells was mediated through oxidative stress, wherein role of iron-mediated LPO was much pronounced in differential cytotoxicity.

  13. [Effects of acidification and alkalinization agents on statins-induced muscle toxicity].

    PubMed

    Kobayashi, Masaki; Hidaka, Kazuhiro; Chisaki, Ikumi; Takahashi, Natsuko; Ogura, Jiro; Itagaki, Shirou; Hirano, Takeshi; Yamaguchi, Hiroaki; Iseki, Ken

    2012-01-01

    The aim of this study was to determine the effects of α-cyano-4-hydroxycinnamic acid (CHC), a lactate efflux inhibitor, and citrate, an alkaline reagent, on statin-induced muscle injury using a human prototypic embryonal rhabdomyosarcoma cell line (RD) as a model of in vitro skeletal muscle and on statin-induced muscle damage in an in vivo study. Statin-induced reduction of cell viability and apoptosis was measured by the 3-(4,5-dimethylthiazol-2-thiazolyl)-2,5-diphenyl tetrazolium bromide (MTT) assay and caspase assay. In an in vivo study, plasma creatine phosphokinase (CPK) level was examined in cerivastatin-treated rats. CHC increased growth inhibition of RD cells induced by cerivastatin, a lipophilic statin, but not these induced by pravastatin, a hydrophilic statin. On the other hand, citrate suppressed cerivastatin-, simvastatin- and atorvastatin-induced reduction of cell viability and caspase activation in RD cells. Moreover, citrate prevented cerivastatin-induced increase in CPK concentration in a concentration-dependent manner. This is first study to evaluate CHC or citrate-induced exacerbation or improvement of statin-induced muscle damage.

  14. Isolation and characterization of new lactam compounds that inhibit lung and colon cancer cells from adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) bran.

    PubMed

    Lee, Ming-Yi; Lin, Huan-You; Cheng, Faiwen; Chiang, Wenchang; Kuo, Yueh-Hsiung

    2008-06-01

    Five active compounds that inhibit cancer cells were isolated from adlay bran (Coix lachryma-jobi L. var. ma-yuen Stapf), and their structures and activities in vitro were characterized. The ethyl acetate-soluble fraction of methanol extracts of adlay bran (ABM-EtOAc) exhibited a stronger anti-proliferative effect on human lung cancer cell A549, human colorectal carcinoma cell HT-29, and COLO 205 than other fractions by MTT (3-4,5-dimethylthiazol-2-yl-2,5-diphenyl-tetrazolium bromide) assay. Assay-guided isolation gave five lactams including three that were previously undocumented; coixspirolactam A (1), coixspirolactam B (2), and coixspirolactam C (3); one isolated from the natural plant for the first time, coixlactam (4); and one known compound, methyl dioxindole-3-acetate (5). Pure active compounds were identified by spectral analysis including IR, 1H and 13C NMR, UV-vis, MS and 2D NMR techniques. All the compounds were tested for their anti-proliferative effect on A549, HT-29 and COLO 205 cells. These compounds showed anti-cancer activities with IC50 values between 28.6 and 72.6microg/mL.

  15. Group 11 complexes with amino acid derivatives: Synthesis and antitumoral studies.

    PubMed

    Ortego, Lourdes; Meireles, Margarida; Kasper, Cornelia; Laguna, Antonio; Villacampa, M Dolores; Gimeno, M Concepción

    2016-03-01

    Gold(I), gold(III), silver(I) and copper(I) complexes with modified amino acid esters and phosphine ligands have been prepared in order to test their cytotoxic activity. Two different phosphine fragments, PPh3 and PPh2py (py=pyridine), have been used. The amino acid esters have been modified by introducing an aromatic amine as pyridine that coordinates metal fragments through the nitrogen atom, giving complexes of the type [M(L)(PR3)](+) or [AuCl3(L)] (L=l-valine-N-(4-pyridylcarbonyl) methyl ester (L1), l-alanine-N-(4-pyridylcarbonyl) methyl ester (L2), l-phenylalanine-N-(4-pyridylcarbonyl) methyl-ester) (L3); M=Au(I), Ag(I), Cu(I), PR3=PPh3, PPh2py). The in vitro cytotoxic activity of metal complexes was tested against four tumor human cell lines and one tumor mouse cell line. A metabolic activity test (3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide, MTT) was used and IC50 values were compared with those obtained for cisplatin. Several complexes displayed significant cytotoxic activities. In order to determine whether antiproliferation and cell death are associated with apoptosis, NIH-3T3 cells were exposed to five selected complexes (Annexin V+ FITC, PI) and analyzed by flow cytometry. These experiments showed that the mechanism by which the complexes inhibit cell proliferation inducing cell death in NIH-3T3 cells is mainly apoptotic.

  16. Antitumor Activities of Ethyl Acetate Extracts from Selaginella doederleinii Hieron In Vitro and In Vivo and Its Possible Mechanism

    PubMed Central

    Li, Juan; Zhao, Ping; Ma, Wen-tao; Feng, Xie-he; Chen, Ke-li

    2015-01-01

    The antitumor activities of ethyl acetate extracts from Selaginella doederleinii Hieron (SD extracts) in vitro and in vivo and its possible mechanism were investigated. HPLC method was developed for chemical analysis. SD extracts were submitted to 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay on different cells, flow cytometry, and RT-PCR analysis using HepG2 cell and antitumor activity in vivo using H-22 xenograft tumor mice. Six biflavonoids from SD extracts were submitted to molecular docking assay. The results showed that SD extracts had considerable antitumor activity in vitro and in vivo without obvious toxicity on normal cells and could induce cell apoptosis. The mechanisms of tumorigenesis and cell apoptosis induced by SD extracts may be associated with decreasing the ratio of bcl-2 and bax mRNA level, activating caspase-3, suppressing survivin, and decreasing the gene expression of COX-2, 5-LOX, FLAP, and 12-LOX mRNA. The main active component in SD extracts is biflavonoids and some exhibited strong interactions with COX-2, 5-LOX, 12-LOX, and 15-LOX. These results offering evidence of possible mechanisms of SD extracts suppress cell proliferation and promote apoptosis and provide the molecular theoretical basis of clinical application of S. doederleinii for cancer therapy. PMID:25866543

  17. Antiviral Activity of Total Flavonoid Extracts from Selaginella moellendorffii Hieron against Coxsackie Virus B3 In Vitro and In Vivo

    PubMed Central

    Yin, Dan; Li, Juan; Lei, Xiang; Liu, Yimei; Yang, Zhanqiu; Chen, Keli

    2014-01-01

    The antiviral activity of total flavonoid extracts from Selaginella moellendorffii Hieron and its main constituents amentoflavone were investigated against coxsackie virus B3 (CVB3). When added during or after viral infection, the extracts and amentoflavone prevented the cytopathic effect (CPE) of CVB3, as demonstrated in a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay, with a 50% inhibitory concentration (IC50) from 19 ± 1.6 to 41 ± 1.2 μg/mL and 25 ± 1.2 to 52 ± 0.8 μg/mL, respectively. KM mice were used as animal models to test the extracts' activity in vivo. Oral administration of the total flavonoid extracts at 300 mg/kg/day significantly reduced mean viral titers in the heart and kidneys as well as mortality after infection for 15 days. The experimental results demonstrate that in vitro and in vivo the model mice infected with CVB3 can be effectively treated by the total flavonoid extracts from Selaginella moellendorffii Hieron. PMID:24963331

  18. Time-Dependent Effect of Encapsulating Alginate Hydrogel on Neurogenic Potential

    PubMed Central

    Razavi, Shahnaz; Khosravizadeh, Zahra; Bahramian, Hamid; Kazemi, Mohammad

    2015-01-01

    Objective Due to the restricted potential of neural stem cells for regeneration of central nervous system (CNS) after injury, providing an alternative source for neural stem cells is essential. Adipose derived stem cells (ADSCs) are multipotent cells with properties suitable for tissue engineering. In addition, alginate hydrogel is a biocompatible polysaccharide polymer that has been used to encapsulate many types of cells. The aim of this study was to assess the proliferation rate and level of expression of neural markers; NESTIN, glial fibrillary acidic protein (GFAP) and microtubule-associated protein 2 (MAP2) in encapsulated human ADSCs (hADSCs) 10 and14 days after neural induction. Materials and Methods In this experimental study, ADSCs isolated from human were cultured in neural induction media and seeded into alginate hydrogel. The rate of proliferation and differentiation of encapsulated cells were evaluated by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay, immunocytoflourescent and realtime reverse transcriptase polymerase chain reaction (RT-PCR) analyzes 10 and 14 days after induction. Results The rate of proliferation of encapsulated cells was not significantly changed with time passage. The expression of NESTIN and GFAP significantly decreased on day 14 relative to day 10 (P<0.001) but MAP2 expression was increased. Conclusion Alginate hydrogel can promote the neural differentiation of encapsulated hADSCs with time passage. PMID:26199909

  19. Structural Characterization and Antioxidative Activity of Low-Molecular-Weights Beta-1,3-Glucan from the Residue of Extracted Ganoderma lucidum Fruiting Bodies

    PubMed Central

    Kao, Pai-Feng; Wang, Shwu-Huey; Hung, Wei-Ting; Liao, Yu-Han; Lin, Chun-Mao; Yang, Wen-Bin

    2012-01-01

    The major cell wall constituent of Ganoderma lucidum (G. lucidum) is β-1,3-glucan. This study examined the polysaccharide from the residues of alkaline-extracted fruiting bodies using high-performance anion-exchange chromatography (HPAEC), and it employed nuclear magnetic resonance (NMR) and mass spectrometry (MS) to confirm the structures. We have successfully isolated low-molecular-weight β-1,3-glucan (LMG), in high yields, from the waste residue of extracted fruiting bodies of G. lucidum. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay evaluated the capability of LMG to suppress H2O2-induced cell death in RAW264.7 cells, identifying that LMG protected cells from H2O2-induced damage. LMG treatment decreased H2O2-induced intracellular reactive oxygen species (ROS) production. LMG also influenced sphingomyelinase (SMase) activity, stimulated by cell death to induce ceramide formation, and then increase cell ROS production. Estimation of the activities of neutral and acid SMases in vitro showed that LMG suppressed the activities of both neutral and acid SMases in a concentration-dependent manner. These results suggest that LMG, a water-soluble β-1,3-glucan recycled from extracted residue of G. lucidum, possesses antioxidant capability against H2O2-induced cell death by attenuating intracellular ROS and inhibiting SMase activity. PMID:22187536

  20. Synthesis, characterization and in vitro anti-cancer evaluation of hesperetin-loaded nanoparticles in human oral carcinoma (KB) cells

    NASA Astrophysics Data System (ADS)

    Gurushankar, K.; Gohulkumar, M.; Rajendra Prasad, N.; Krishnakumar, N.

    2014-03-01

    Hesperetin (HET), a naturally occurring plant bioflavonoid present in citrus fruits, possesses potential anti-inflammatory and anti-carcinogenic activities but poor aqueous solubility limits its applications. To improve its applicability in cancer therapy, hesperetin was encapsulated in Eudragit® E (EE) 100 nanoparticles in the presence of polyvinyl alcohol (PVA) as a stabilizer and its anticancer efficacy in oral carcinoma (KB) cells was studied. Hesperetin-loaded nanoparticles (HETNPs) were prepared by nanoprecipitation method and characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), Fourier transform infrared (FT-IR) spectroscopy, differential scanning calorimetry (DSC), and x-ray diffraction (XRD). The results thus displayed that the prepared nanoparticles showed a particle size in the range from 55 to 180 nm. The encapsulation efficiency of hesperetin was 83.4% obtained by UV spectroscopy. The in vitro release kinetics of hesperetin under physiological condition show initial rapid release followed by slow and sustained release. 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay revealed higher cytotoxic efficacy of HETNPs than native hesperetin in KB cells. Further, it has been found that reactive oxygen species (ROS) generation, DNA damage and apoptotic indices in HETNPs treated cells are greater than those in native hesperetin treatment. Hence these findings demonstrate that HETNPs could be a potentially useful drug delivery system to produce better hesperetin therapeutics of cancers.

  1. Acacia honey and chrysin reduce proliferation of melanoma cells through alterations in cell cycle progression.

    PubMed

    Pichichero, Elena; Cicconi, Rosella; Mattei, Maurizio; Muzi, Marco Gallinella; Canini, Antonella

    2010-10-01

    Honey has long been used in medicine for different purposes. Only recently, however, its antioxidant property and preventive effects against different diseases, such as cancer, have been highlighted. Chrysin (5,7-dihydroxyflavone) is a natural flavone commonly found in acacia honey. It has previously been shown to be an anti-tumor agent. In this study, we investigated the antiproliferative role of honey or chrysin on human (A375) and murine (B16-F1) melanoma cell lines. The results of the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and the trypan blue exclusion test showed that both the tested compounds were able to induce an antiproliferative effect on melanoma cells in a dose- and time-dependent manner. Flow cytometry analysis indicated that cytotoxicity induced by honey or chrysin was mediated by G(0)/G(1) cell cycle arrest and induction of hyperploid progression. Our results suggest that the anti-proliferative effects of honey are due mainly to the presence of chrysin. Chrysin may therefore be considered a potential candidate for both cancer prevention and treatment. Further investigation is needed to validate the contribution of chrysin in tumor therapy in vivo.

  2. Synthesis, characterization and biological evaluation of ruthenium flavanol complexes against breast cancer.

    PubMed

    Singh, Ashok Kumar; Saxena, Gunjan; Sahabjada; Arshad, M

    2017-03-01

    Four Ru(II) DMSO complexes (M1R-M4R) having substituted flavones viz. 3-Hydroxy-2-(4-methoxyphenyl)-4H-chromen-4-one (HL1), 3-Hydroxy-2-(4-nitrophenyl)-4H-chromen-4-one (HL2), 3-Hydroxy-2-(4-dimethylaminophenyl)-4H-chromen-4-one (HL3) and 3-Hydroxy-2-(4-chlorophenyl)-4H-chromen-4-one (HL4) were synthesized and characterized by elemental analysis, IR, UV-Vis, (1)H NMR spectroscopies and ESI-MS. The molecular structures of the complexes were investigated by integrated spectroscopic and computational techniques (DFT). Both ligands as well as their complexes were screened for anticancer activities against breast cancer cell lines MCF-7. Cytotoxicity was assayed by MTT [3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide] assay. All ligands and their complexes exhibited significant cytotoxic potential of 5-40μM concentration at incubation period of 24h. The cell cytotoxicity increased significantly in a concentration-dependent manner. In this series of compounds, HL2 (IC50 17.2μM) and its complex M2R (IC50 16μM) induced the highest cytotoxicity.

  3. In vitro cellular uptake and cytotoxic effect of functionalized nickel nanoparticles on leukemia cancer cells.

    PubMed

    Guo, Dadong; Wu, Chunhui; Li, Xiaomao; Jiang, Hui; Wang, Xuemei; Chen, Baoan

    2008-05-01

    Nickel nanoparticles (Ni NPs) have been applied in a wide range of areas because of their unique structure and properties such as catalysts, high-density magnetic recording media and others. However, little effort has been paid to their biological application and the concrete effect of Ni NPs on biological systems is still unknown. In this study, the possibility of the utilization of the magnetic Ni NPs in cancer cell studies was explored and the effects of the Ni NPs capped with positively charged tetraheptylammonium on leukemia K562 cells in vitro were investigated. Our observations of optical microscopy, atomic force microscopy (AFM) and scanning electron microscopy (SEM) studies indicate that the morphological changes of cancer cells induced by Ni NPs could be apparently observed. The results of 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide (MTT) assay, DNA fragmentation and flow cytometry studies demonstrate that the Ni NPs could exert cytotoxicity to leukemia K562 cells at high concentration, and subsequently induce both apoptosis and necrosis of target cancer cells, whilst it had little impact on target cells when at low concentration. Meanwhile, functionalized Ni NPs with positively charged groups could enhance the permeability of cell membrane and facilitate the cellular uptake of outer target molecules into cancer cells. These findings reveal the potential mechanism of Ni NPs to target cancer cells which could induce the cytotoxicity to leukemia cancer cells and suggest the possibility for applications of the Ni NPs in related clinical and biomedical areas.

  4. In vitro biocompatibility of chitosan porous skin regenerating templates (PSRTs) using primary human skin keratinocytes.

    PubMed

    Lim, C K; Yaacob, N S; Ismail, Z; Halim, A S

    2010-04-01

    Biopolymer chitosan (beta-1,4-d-glucosamine) comprises the copolymer mixture of N-acetylglucosamine and glucosamine. The natural biocompatibility and biodegradability of chitosan have recently highlighted its potential use for applications in wound management. Chemical and physical modifications of chitosan influence its biocompatibility and biodegradability, but it is unknown as to what degree. Hence, the biocompatibility of the chitosan porous skin regenerating templates (PSRT 82, 87 and 108) was determined using an in vitro toxicology model at the cellular and molecular level on primary normal human epidermal keratinocytes (pNHEK). Cytocompatibility was accessed by using a 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl tetrazolium bromide (MTT) assay from 24 to 72h. To assess the genotoxicity of the PSRTs, DNA damage to the pNHEK was evaluated by using the Comet assay following direct contact with the various PSRTs. Furthermore, the skin pro-inflammatory cytokines TNF-alpha and IL-8 were examined to evaluate the tendency of the PSRTs to provoke inflammatory responses. All PSRTs were found to be cytocompatible, but only PSRT 108 was capable of stimulating cell proliferation. While all of the PSRTs showed some DNA damage, PSRT 108 showed the least DNA damage followed by PSRT 87 and 82. PSRT 87 and 82 induced a higher secretion of TNF-alpha and IL-8 in the pNHEK cultures than did PSRT 108. Hence, based on our experiments, PSRT 108 is the most biocompatible wound dressing of the three tested.

  5. Kale extract increases glutathione levels in V79 cells, but does not protect them against acute toxicity induced by hydrogen peroxide.

    PubMed

    Fernandes, Fátima; Sousa, Carla; Ferreres, Federico; Valentão, Patrícia; Remião, Fernando; Pereira, José A; Andrade, Paula B

    2012-05-07

    This study aims to evaluate the antioxidant potential of extracts of Brassica oleracea L. var. acephala DC. (kale) and several materials of Pieris brassicae L., a common pest of Brassica cultures using a cellular model with hamster lung fibroblast (V79 cells) under quiescent conditions and subjected to H₂O₂ induced oxidative stress. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and glutathione was determined by the 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB)-oxidized glutathione (GSSG) reductase recycling assay. The phenolic composition of the extracts was also established by HPLC-DAD. They presented acylated and non acylated flavonoid glycosides, some of them sulfated, and hydroxycinnamic acyl gentiobiosides. All extracts were cytotoxic by themselves at high concentrations and failed to protect V79 cells against H₂O₂ acute toxicity. No relationship between phenolic composition and cytotoxicity of the extracts was found. Rather, a significant increase in glutathione was observed in cells exposed to kale extract, which contained the highest amount and variety of flavonoids. It can be concluded that although flavonoids-rich extracts have the ability to increase cellular antioxidant defenses, the use of extracts of kale and P. brassicae materials by pharmaceutical or food industries, may constitute an insult to health, especially to debilitated individuals, if high doses are consumed.

  6. Rhizoma Paridis Saponins Induces Cell Cycle Arrest and Apoptosis in Non-Small Cell Lung Carcinoma A549 Cells

    PubMed Central

    Zhang, Jue; Yang, Yixi; Lei, Lei; Tian, Mengliang

    2015-01-01

    Background As a traditional Chinese medicine herb, Chonglou (Paris polyphylla var. chinensis) has been used as anticancer medicine in China in recent decades, as it can induce cell cycle arrest and apoptosis in numerous cancer cells. The saponins extract from the rhizoma of Chonglou [Rhizoma Paridis saponins (RPS)] is known as the main active component for anticancer treatment. However, the molecular mechanism of the anticancer effect of RPS is unknown. Material/Methods The present study evaluated the effect of RPS in non-small-cell lung cancer (NSCLC) A549 cells using the 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (MTT) assay and flow cytometry. Subsequently, the expression of several genes associated with cell cycle and apoptosis were detected by reverse transcription-quantitative polymerase chain reaction (qRT-PCR) and Western blotting. Results RPS was revealed to inhibit cell growth, causing a number of cells to accumulate in the G 1 phase of the cell cycle, leading to apoptosis. In addition, the effect was dose-dependent. Moreover, the results of qRT-PCR and Western blotting showed that p53 and cyclin-dependent kinase 2 (CDK2) were significantly downregulated, and that BCL2, BAX, and p21 were upregulated, by RPS treatment. Conclusions We speculated that the RPS could act on a pathway, including p53, p21, BCL2, BAX, and CDK2, and results in G1 cell cycle arrest and apoptosis in NSCLC cells. PMID:26311066

  7. Potential of a soluble tetrazolium/formazan assay for the evaluation of filarial viability.

    PubMed

    Comley, J C; Turner, C H

    1990-04-01

    Using female Acanthocheilonema viteae we have investigated the bioreduction of the tetrazolium reagent XTT (2,3-bis(2-methoxy-4-nitro-sulphonyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide). Unlike the formazan formed by other tetrazolium salts, that derived from XTT readily diffuses out of A. viteae in vitro. Formazan formation can therefore be quantified by direct absorbance reading of the incubation medium, eliminating the need for a DMSO solubilization step. Optimum assay conditions involved a 4 h incubation, in the presence of the electron coupling agent phenazine methosulphate (PMS). Repeat 4 h incubations with XTT-PMS were well tolerated by worms for 5 consecutive days. This confirmed the low toxicity of XTT formazan and its usefulness in the semi-continuous assessment of filarial viability. In comparison to our previously reported MTT (3-(4, 5 dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide)-reduction assay XTT-PMS reduction showed comparable drug sensitivity and accuracy, however XTT-PMS appears to be at least 10-15 times less efficiently reduced by A. viteae females. A possible application of the XTT assay using female Onchocerca volvulus is discussed.

  8. Effects of naproxen on cell proliferation and genotoxicity in MG-63 osteosarcoma cell line.

    PubMed

    Correia, Isabel; Arantes-Rodrigues, Regina; Pinto-Leite, Rosário; Gaivão, Isabel

    2014-01-01

    The purpose of this study was to determine the efficacy of naproxen, a nonsteroidal anti-inflammatory drug, on the MG-63 human osteosarcoma cell line. MG-63 cells were exposed to naproxen in a wide range of concentrations of 0.03, 0.05, 0.1, 0.42, 0.83, and 1.67 mg/ml for 72 h. The activity of naproxen was assessed by the following assays: cell morphology; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method; terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay; comet assay; and acridine orange and monodansylcadaverine (MDC) staining. Naproxen exerted a significant inhibitory effect on MG-63 cell proliferation, in a concentration-dependent manner, in all treatment groups compared with untreated cells. An increase in frequency of DNA damage, apoptotic bodies, apoptotic cells, and autophagic vacuoles was observed in MG-63-treated cells. Although future studies are needed, these findings suggest that naproxen may lead to improvements in treatment of patients with osteosarcoma.

  9. Citotoxicity evaluation of three dental adhesives on vero cells in vitro

    PubMed Central

    Catunda, Raisa-Queiroz; Vieira, Jeymesson-Raphael-Cardoso; de Oliveira, Erwelly-Barros; da Silva, Eliete-Cavalcanti; Brasil, Veruska-Lima-Moura

    2017-01-01

    Background To evaluate, in vitro, the potential cytotoxicity of three different dental adhesives systems (Adper Single Bond 2 -SB, Silorane System Adhesive Bond -SSAB and Single Bond Universal -SBU) on cultivated Vero cells after different contact times. Material and Methods The cells were cultured in a concentration of 2 x 105 cells/mL for 24h and grown to sub-confluent monolayers. VERO cells were exposed to 25µl of conditioned extracts obtained from 24h, 48h and 72h immersion of adhesive samples in culture medium (DMEM), immediately after polymerization. Fresh DMEM was used as negative control. Cell metabolism was evaluated by the MTT assay (3-(4,5-dimethylthiazol-2-yl)-2, 5diphenyl-tetrazolium bromide). The data were analyzed statistically by ANOVA, considering a significance of 5%. Results The values of cell viability ranged from 94.2% at 72h (SBU) to 109.6% at 48h (SB). The mean percentage of viability after exposure to the extracts of SB, SSAB and SBU were 103.2%, 100.63% and 97.43%, respectively. There was no statistically significant difference (p= 0.342) between the experimental and negative control groups. Conclusions At all exposure times, all adhesives tested in this study presented no cytotoxicity to Vero cells in vitro. Key words:Biocompatibility, cytotoxicity, dental adhesives, Vero cells. PMID:28149465

  10. Flavonoid C-glucosides Derived from Flax Straw Extracts Reduce Human Breast Cancer Cell Growth In vitro and Induce Apoptosis.

    PubMed

    Czemplik, Magdalena; Mierziak, Justyna; Szopa, Jan; Kulma, Anna

    2016-01-01

    Flax straw of flax varieties that are grown for oil production is a by product which represents a considerable biomass source. Therefore, its potential application for human use is of high interest. Our research has revealed that flax straw is rich in flavonoid C-glucosides, including vitexin, orientin, and isoorientin. The objective of this study was to evaluate the cytotoxicity and possible proapoptotic effect of flax straw derived C-glucosides of flavonoids in the human breast adenocarcinoma cell line (MCF-7). The effects of flax straw derived flavonoid C-glucosides on cell proliferation of MCF-7 cells were evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) and sulforhodamine B assays. The expression of apoptosis-related genes was assessed by real-time PCR. Our data revealed that flax C-glucosides as well as pure compounds are cytotoxic toward MCF-7 cells and inhibit their proliferation. Moreover, the induction of apoptosis was correlated with the changes in the mRNA level of pro-apoptotic genes. Increased expression of bax and caspase-7, -8, and -9 and decreased mRNA expression of bcl-2 was observed, whereas the mRNA levels of p53 and mdm2 were not altered. These results clearly demonstrated that flax straw metabolites effectively induced growth inhibition and apoptosis in human breast adenocarcinoma cells.

  11. Quercetin protects hamster spermatogenic cells from oxidative damage induced by diethylstilboestrol.

    PubMed

    Li, G; Ma, Aituan; Shi, W; Zhong, Xiuhui

    2010-10-01

    Quercetin has been reported to be an efficient antioxidant which protects chicken spermatogonial cells from oxidative damage through increasing intracellular antioxidants and decreasing lipid peroxidation. Exposure to diethylstilboestrol (DES) could cause reproductive damage in males, which is associated with oxidative stress. This study was conducted to investigate the protective effects of quercetin on DES-induced oxidative damage in cultured hamster spermatogenic cells. The cells were treated with different concentrations of DES, and their growth status was observed under inverted microscope. The viability of spermatogenic cells was detected by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT). The contents of superoxide dismutase (SOD) in supernatants and glutathione peroxidase (GSH-Px) in cells were detected with spectrophotography. The results showed that quercetin significantly inhibited the DES-induced damage on spermatogenic cells, with the exception of the low-dose group in which no significant difference was observed. The cell survival rate increased significantly in the middle- and high-dose groups. The contents of SOD and GSH-Px were significantly elevated after medication with quercetin (P < 0.01). It can be concluded that quercetin protects spermatogenic cells against DES-induced oxidative damage through increasing intracellular antioxidants and decreasing lipid peroxidation. Quercetin plays a very important role in ameliorating reproductive toxicity induced by environmental oestrogens.

  12. Flavonoid C-glucosides Derived from Flax Straw Extracts Reduce Human Breast Cancer Cell Growth In vitro and Induce Apoptosis

    PubMed Central

    Czemplik, Magdalena; Mierziak, Justyna; Szopa, Jan; Kulma, Anna

    2016-01-01

    Flax straw of flax varieties that are grown for oil production is a by product which represents a considerable biomass source. Therefore, its potential application for human use is of high interest. Our research has revealed that flax straw is rich in flavonoid C-glucosides, including vitexin, orientin, and isoorientin. The objective of this study was to evaluate the cytotoxicity and possible proapoptotic effect of flax straw derived C-glucosides of flavonoids in the human breast adenocarcinoma cell line (MCF-7). The effects of flax straw derived flavonoid C-glucosides on cell proliferation of MCF-7 cells were evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) and sulforhodamine B assays. The expression of apoptosis-related genes was assessed by real-time PCR. Our data revealed that flax C-glucosides as well as pure compounds are cytotoxic toward MCF-7 cells and inhibit their proliferation. Moreover, the induction of apoptosis was correlated with the changes in the mRNA level of pro-apoptotic genes. Increased expression of bax and caspase-7, -8, and -9 and decreased mRNA expression of bcl-2 was observed, whereas the mRNA levels of p53 and mdm2 were not altered. These results clearly demonstrated that flax straw metabolites effectively induced growth inhibition and apoptosis in human breast adenocarcinoma cells. PMID:27630565

  13. Mitochondria Related Pathway Is Essential for Polysaccharides Purified from Sparassis crispa Mediated Neuro-Protection against Glutamate-Induced Toxicity in Differentiated PC12 Cells

    PubMed Central

    Hu, Shuang; Wang, Di; Zhang, Junrong; Du, Mengyan; Cheng, Yingkun; Liu, Yan; Zhang, Ning; Wang, Di; Wu, Yi

    2016-01-01

    The present study aims to explore the neuro-protective effects of purified Sparassis crispa polysaccharides against l-glutamic acid (l-Glu)-induced differentiated PC12 (DPC12) cell damages and its underlying mechanisms. The Sparassis crispa water extract was purified by a DEAE-52 cellulose anion exchange column and a Sepharose G-100 column. A fraction with a molecular weight of 75 kDa and a diameter of 88.9 nm, entitled SCWEA, was obtained. SCWEA was identified with a triple helix with (1→3)-linked Rha in the backbone, and (1→2) linkages and (1→6) linkages in the side bone. Our results indicated that the pre-treatment of DPC12 cells with SCWEA prior to l-Glu exposure effectively reversed the reduction on cell viability (by 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay) and reduced l-Glu-induced apoptosis (by Hoechst staining). SCWEA decreased the accumulation of intracellular reactive oxygen species, blocked Ca2+ influx and prevented depolarization of the mitochondrial membrane potential in DPC12 cells. Furthermore, SCWEA normalized expression of anti-apoptotic proteins in l-Glu-explored DPC12 cells. These results suggested that SCWEA protects against l-Glu-induced neuronal apoptosis in DPC12 cells and may be a promising candidate for treatment against neurodegenerative disease. PMID:26821016

  14. Inhibition of bactericidal activity by pentachlorophenol in two phagocyte populations from Fundulus heteroclitus

    SciTech Connect

    Roszell, L.E.; Anderson, R.S.

    1994-12-31

    The effects of pentachlorophenol (PCP) on the bactericidal activity of pronephritic phagocytes were studied in an estuarine fish, Fundulus heteroclitus. Following in vitro exposure to sublethal doses of PCP, macrophages and eosinophils were challenged with Listonella anguillarum, the bacterium responsible for vibriosis in marine and freshwater fish. Quantification of surviving bacteria was based on the reduction of MTT (3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide). Bacteridical activity was inhibited at PCP concentrations greater than 5 ppm in both leukocyte populations; at 20 ppm bactericidal activity was essentially eliminated. The primary cellular mechanisms of bactericidal activity in these cells are phagocytosis and the phagocytically induced production of reactive oxygen intermediates (ROIs) including superoxide (O{sub 2}{sup {minus}}) and hydrogen peroxide (H{sub 2}O{sub 2}). Previous experiments have shown that these activities are inhibited at similar concentrations of PCP. These results indicate that the suppression of phagocytosis and the subsequent oxidative burst is responsible for the reduced killing seen in the current experiments. Nonspecific immune activities of phagocytic cells such as macrophages and eosinophils act as a first line of defense against invading pathogens; the suppression of these functions could ultimately lead to decreased resistance to infectious disease.

  15. Effects of salinomycin on human bone marrow-derived mesenchymal stem cells in vitro.

    PubMed

    Scherzed, A; Hackenberg, S; Froelich, K; Rak, K; Technau, A; Radeloff, A; Nöth, U; Koehler, C; Hagen, R; Kleinsasser, N

    2013-04-26

    Various hypotheses on the origin of cancer stem cells (CSCs) exist, including that CSCs develop from transformed human bone marrow mesenchymal stem cells (hBMSC). Since the polyether antibiotic salinomycin selectively kills CSCs, the present study aims to elucidate the effects of salinomycin on normal hBMSC. The immunophenotype of hBMSC after salinomycin exposure was observed by flow cytometry. The multi-differentiation capacity of hBMSC was evaluated by Oil Red O and van Kossa staining. Cytotoxic effects of salinomycin were monitored by the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] (MTT) assay. Furthermore, spheroid formation and migration capacity were assessed. There were no differences in the immunophenotype and multi-differentiation capacity of hBMSC induced by salinomycin treatment. Cytotoxic effects were observed at concentrations of 30 μM and above. Neither the migration capability nor the ability to form spheroids was affected. Essential functional properties of hBMSC were unaffected by salinomycin. However, dose-dependent cytotoxicity effects could be observed. Overall, low dose salinomycin showed no negative effects on hBMSC. Since mesenchymal stem cells from various sources respond differently, further in vitro studies are needed to clarify the effect of salinomycin on tissue-specific stem cells.

  16. The MLN4924 inhibitor exerts a neuroprotective effect against oxidative stress injury via Nrf2 protein accumulation.

    PubMed

    Andérica-Romero, Ana Cristina; Hernández-Damián, Jacqueline; Vázquez-Cervantes, Gustavo Ignacio; Torres, Ismael; Pedraza-Chaverri, José

    2016-08-01

    It was explored the cytoprotective and antioxidant effect of MLN4924, a specific inhibitor of Nedd8-activating enzyme (NAE), against hydrogen peroxide (H2O2)-induced damage in cerebellar granule neurons (CGNs). Primary cultures of CGNs were exposed to H2O2 after preincubation with MLN4924. The compounds were removed, and CGNs were incubated in culture medium for 24h in order to determine cell viability by 3-[4,5-dimethylthiazol-2-yl)]-2,5-diphenyl-tetrazolium bromide (MTT) and fluorescein diacetate (FDA) assays. It was demonstrated that MLN4924 remarkably attenuated H2O2-induced cell damage. Meanwhile reactive oxygen species (ROS) production was evaluated with the fluorescent probe dihydroethidium (DHE). Interestingly H2O2-induced ROS production was inhibited by pretreatment with MLN4924. MLN4924 treatment in CGNs resulted in nuclear factor E2-related factor 2 (Nrf2) protein accumulation. Intriguingly this effect was observed in the cytosolic and nuclear compartments of the CGNs. The cytoprotective effect of MLN4924 was associated with its ability to diminish ROS production induced by H2O2 and the accumulation of Nrf2 protein levels in the cytoplasm and nucleus of the CGNs.

  17. Inhibition of AGS Cancer Cell Proliferation following siRNA-Mediated Downregulation of VEGFR2

    PubMed Central

    Zarei Mahmudabadi, Ali; Masoomi Karimi, Masoomeh; Bahabadi, Majid; Bagheri Hoseinabadi, Zahra; JafariSani, Moslem; Ahmadi, Reza

    2016-01-01

    Objective Vascular endothelial growth factor (VEGF) and VEGF receptors (VEGFRs) play important roles in angiogenesis of different developmental mechanisms such as wound healing, embryogenesis and diseases, including different types of cancer. VEGFR2 is associated with cell proliferation, migration, and vascular permeability of endothelial cells. Blocking VEGF and its receptors is suggested as a therapeutic approach to prevent tumor growth. In this study, we aim to block VEGF signaling via small interfering RNA (siRNA) inhibition of VEGFR2. Materials and Methods In this experimental study, we used the RNA interference (RNAi) mechanism to suppress expression of the VEGFR2 gene. We conducted the 3-(4,5-di- methylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, real-time polymerase chain reaction (PCR), Western blot, and flow cytometry analyses of VEGFR2 expression. Results Real-time PCR and Western blot results showed that VEGFR2 expression significantly downregulated. This suppression was followed by inhibition of cell prolifera- tion, reduction of viability, and induction of apoptosis in the cancer cells. Conclusion These findings suggest that VEGFR2 has a role in cell proliferation and tumor growth. Accordingly, it is suggested that VEGFR2 can be a therapeutic target for controlling tumor growth and proliferation. PMID:27602320

  18. Inhibition activities of polysaccharide (RG4-1) from Gentiana rigescens against RSV.

    PubMed

    Li, Xin; Zhou, Jing-Yu; Yang, Qing-Wei; Chen, Yang; Piao, Ying-Ai; Li, Hong-Yuan

    2011-06-01

    Respiratory syncytial virus (RSV) is the most important cause of lower respiratory tract infection in infants and young children. With the emergence of drug-resistant strains of RSV, new antiviral agents are needed urgently. Gentiana rigescens is a kind of Chinese herb, belonging to Gentianaceae, which has long been used as a folk medicine for curing inflammation, bacterial infection, viral infection, and so on. In this research, polysaccharide designated RG4-1 was isolated from G. rigescens by hot water extraction, ethanol precipitation, and macroreticular adsorbing resin column chromatography, and its antiviral activity, cytotoxicity, and possible antiviral mechanisms were assayed by cytopathogenic effect inhibition assay, 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and plaque reduction assay. RG4-1 was a fructose-binding lectin. In host cell cultures, RG4-1 was found to be an effective antiviral component against RSV. It showed good inhibitory effect against RSV when it was added 2 h after virus infection with 50% effective concentration of 12.86 μg/ml. RG4-1 also displayed its direct inactivation, attachment inhibition effect, and penetration inhibition effect against RSV. A time-dependent experiment was set up to confirm that RG4-1 blocked RSV infection at early stages of the infection. But RG4-1 seemed to be ineffective against intracellular virus and viral biosynthesis.

  19. Integration of Rabbit Adipose Derived Mesenchymal Stem Cells to Hydroxyapatite Burr Hole Button Device for Bone Interface Regeneration

    PubMed Central

    Gayathri, Viswanathan; Harikrishnan, Varma; Mohanan, Parayanthala Valappil

    2016-01-01

    Adipose Derived Mesenchymal Stem Cells, multipotent stem cells isolated from adipose tissue, present close resemblance to the natural in vivo milieu and microenvironment of bone tissue and hence widely used for in bone tissue engineering applications. The present study evaluates the compatibility of tissue engineered hydroxyapatite burr hole button device (HAP-BHB) seeded with Rabbit Adipose Derived Mesenchymal Stem Cells (ADMSCs). Cytotoxicity, oxidative stress response, apoptotic behavior, attachment, and adherence of adipose MSC seeded on the device were evaluated by scanning electron and confocal microscopy. The results of the MTT (3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide) assay indicated that powdered device material was noncytotoxic up to 0.5 g/mL on cultured cells. It was also observed that oxidative stress related reactive oxygen species production and apoptosis on cell seeded device were similar to those of control (cells alone) except in 3-day period which showed increased reactive oxygen species generation. Further scanning electron and confocal microscopy indicated a uniform attachment of cells and viability up to 200 μm deep inside the device, respectively. Based on the results, it can be concluded that the in-house developed HAP-BHB device seeded with ADMSCs is nontoxic/safe compatible device for biomedical application and an attractive tissue engineered device for calvarial defect regeneration. PMID:26880922

  20. Petiveria alliacea extracts uses multiple mechanisms to inhibit growth of human and mouse tumoral cells

    PubMed Central

    Urueña, Claudia; Cifuentes, Claudia; Castañeda, Diana; Arango, Amparo; Kaur, Punit; Asea, Alexzander; Fiorentino, Susana

    2008-01-01

    Background There is ethnopharmacological evidence that Petiveria alliacea can have antitumor activity; however, the mechanism of its cytotoxic activity is not well understood. We assessed multiple in vitro biological activities of an ethyl acetate soluble plant fraction over several tumor cell lines. Methods Tumor cell lines were evaluated using the following tests: trypan blue exclusion test, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide], flow cytometry, cytoskeleton organization analysis, cell cycle, mitochondria membrane depolarization, clonogenicity test, DNA fragmentation test and differential protein expression by HPLC-Chip/MS analysis. F4 fraction characterization was made by HPLC-MS. Results Petiveria alliacea fraction characterized by de-replication was found to alter actin cytoskeleton organization, induce G2 cell cycle arrest and cause apoptotic cell death in a mitochondria independent way. In addition, we found down regulation of cytoskeleton, chaperone, signal transduction proteins, and proteins involved in metabolic pathways. Finally up regulation of proteins involved in translation and intracellular degradation was also observed. Conclusion The results of this study indicate that Petiveria alliacea exerts multiple biological activities in vitro consistent with cytotoxicity. Further studies in animal models are needed but Petiveria alliacea appears to be a good candidate to be used as an antitumor agent. PMID:19017389

  1. Antiangiogenic and antiproliferative effects of black pomegranate peel extract on melanoma cell line

    PubMed Central

    Dana, N.; Javanmard, Sh. Haghjooy; Rafiee, L.

    2015-01-01

    In the present study possible effects of black pomegranate peel extract (PPE) on the B16F10 melanoma cells proliferation and Human Umbilical Vein Endothelial Cells (HUVECs) angiogenesis were investigated. PPE was added into the cell lines (B16F10 and HUVECs) media with different concentrations (10–450 μg/ml). After 48 h, the cell survival was measured by 3-(Dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Angiogenesis was investigated by matrigel assay (PPE (200, 300, 400 μg/ml)); HUVECs, vascular endothelial growth factor (VEGF) mRNA expression was detected by quantitative reverse transcriptase–polymerase chain reaction (QRT-PCR) assay. VEGF concentration in culture medium of HUVECs was determined by enzyme-linked immunosorbent assay (ELISA). PPE had positive anti proliferative effect on melanoma cells in a dose-dependent manner, but not on HUVECs. The matrigel assay results indicated that PPE significantly inhibited length, size and junction of the tube like structures (P<0.05). VEGF mRNA expression and concentration levels in culture medium of PPE treated HUVECs reduced significantly in a concentration-dependent manner (P<0.05). Simultaneous inhibition of melanoma cell proliferation and angiogenesis proposed that, PPE can be a good candidate against melanoma development. Based on the results, PPE could effectively suppress angiogenesis potentially through a VEGF dependent mechanism. Further studies are needed to confirm these results. PMID:26487888

  2. Rab14 Act as Oncogene and Induce Proliferation of Gastric Cancer Cells via AKT Signaling Pathway

    PubMed Central

    Zhao, Zhenghao; Li, Qian; Zhou, Kaiyue; Zhao, Lingyu; Wang, Lumin; Yang, Juan; Huang, Chen

    2017-01-01

    Rab14 is a member of RAS oncogene family, and its dysfunction has been reported to be involved in various types of human cancer. However, its expression and function were still unclear in gastric cancer. The aim of this study was to investigate the function and mechanism of Rab14 in gastric cancer cell lines. Quantitative real-time PCR (qRT-PCR) was performed in 17 gastric adenocarcinoma tissues and 4 cell lines to detect the expression of Rab14. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT), colony formation and flow cytometry assays were employed to determine the proliferative ability, cell cycle transition and apoptosis in vitro in BGC-823 or SGC-7901 cells. Western blot was performed to investigate the pathways and mechanism of Rab14 regulation. In this study, we show that Rab14 presents a significant up-regulated expression among the paired tissue samples and cell lines in gastric cancer. When we overexpressed Rab14 in SGC-7901 cells or silenced Rab14 in BGC-823 cells, we found that Rab14 could modify cell growth, cell cycle or apoptosis, which accompanied with an obvious regulation of CCND1, CDK2 and BAX involving in AKT signaling pathway. In conclusion, this study provides a new evidence on that Rab14 functions as a novel tumor oncogene and could be a potential therapeutic target in gastric cancer. PMID:28107526

  3. Protective effect of deferricoprogen isolated from Monascus purpureus NTU 568 on citrinin-induced apoptosis in HEK-293 cells.

    PubMed

    Hsu, Li-Chuan; Hsu, Ya-Wen; Liang, Yu-Han; Lin, Zhi-Hu; Kuo, Yao-Haur; Pan, Tzu-Ming

    2012-08-15

    Monascus species have traditionally been used in Asian food, with rice as their fermentation substrate. Red mold rice (RMR) contains citrinin, a nephrotoxic agent capable of exerting oxidative stress and cellular apoptosis. We investigated the components in RMR that could minimize the adverse effects of citrinin. Combining chemical separations and bioactivity assays, we identified an antioxidative component called deferricoprogen (DFC) in the fermented rice of Monascus purpureus NTU 568. The DFC structure was confirmed by nuclear magnetic resonance (NMR) and mass spectra analysis. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical-scavenging activity of DFC was similar to that of vitamin E. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and flow cytometric analysis showed the effect of DFC and citrinin on cell viability and cell cycle. DFC was found to be protective against the cytotoxicity and cell death induced by citrinin on human embryonic kidney (HEK-293) cells. DFC also demonstrated anti-apoptotic property in preventing citrinin-induced apoptosis.

  4. Anti-proliferative and Apoptotic Effects of Dendrosomal Farnesiferol C on Gastric Cancer Cells.

    PubMed

    Aas, Zohreh; Babaei, Esmaeil; Hosseinpour Feizi, Mohammad Ali; Dehghan, Gholamreza

    2015-01-01

    Farnesiferol C is a natural compound with various anti-cancer properties that belongs to the class of sesquiterpene coumarins. However, the low bioavailability of farnesiferol C limits its therapeutic potential. Here, we overcame this problem utilizing dendrosome nano-particles and evaluated the anti-cancer effect of dendrosomal farnesiferol C (DFC) on the AGS gastric cancer cell line. The 3-(4,5-dimethyl-thiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and reverse transcriptase-polymerase chain reaction (RT-PCR) were respectively used to detect the anti-proliferative properties of DFC and expression ratio of Bax/Bcl-2 as a hallmark of apoptosis. Compared to the void farnesiferol C (FC), our data showed that DFC significantly suppresses the proliferation of AGS cells in a time- and dose-dependent manner (P<0.01). Also, DFC meaningfully increased the expression ratio of Bax/Bcl-2 in AGS cells (P<0.01). The findings demonstrate that our nano-based formulation of farnesiferol C could be considered as a potential therapeutic agent in cancer targeting.

  5. In vitro evaluation of acute cytotoxicity of human chemically treated allografts.

    PubMed

    Dufrane, D; Cornu, O; Verraes, T; Schecroun, N; Banse, X; Schneider, Y J; Delloye, C

    2001-01-10

    In order to minimize the risk of contamination associated with tissue transplantation, tissue banks commonly chemically treat the tissues whenever possible. As viral inactivation uses agents lethal to microorganisms, it is imperative to assure that chemically inactivated tissue remains biocompatible. In vitro assays can be an effective means to assess the acute cytotoxicity of chemically treated human allografts. We have used different types of cells cultured in the presence of treated tissue extract. A standard cell line, a human fibroblast (WI38), which was the same for all the samples, was chosen. In addition, as the banked tissues (bone and fascia lata) were prepared to be used in bone or as a dura mater substitute, two other cell types were also used: an osteoblastic cell line (SaOS-2) and a neuronal cell line (Neuro 2A). Cytotoxic assessment was performed by qualitative evaluation of cell morphology based on confluence, granulation, vacuolization and swelling analysis. In addition, quantitative methods based on the use of neutral red (NR) and 3- (4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) were assayed. Qualitative and quantitative evaluation of fascia lata and bone extracts did not show deleterious effects on cell cultures. These results show that in vitro methods can be appropriate to select a non-toxic procedure before it is used in the human body and that several strong chemical treatments can result in a tissue suitable for human.

  6. The effect of antioxidants on glycated albumin-induced cytotoxicity in bovine retinal pericytes.

    PubMed

    Kim, Jaetaek; Kim, Kwang-seok; Shinn, Jong-wook; Oh, Yeon-sahng; Kim, Heung-tae; Jo, Inho; Shinn, Soon-hyun

    2002-04-12

    Loss of retinal pericytes is the initial deficit in the early stage of diabetic retinopathy. Glycated albumin (GA) forms under hyperglycemic conditions and exists in the retinal blood vessels of diabetic patients with retinopathy. In this study, using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) reduction test, we investigated whether GA induces cytotoxicity in cultured bovine retinal pericytes, and whether the antioxidants, l-ascorbic acid, Trolox, and probucol, provide any protection from GA-mediated cytotoxicity. GA induced pericyte death in a dose-dependent manner. With increasing time, GA-induced cytotoxicity also increased despite no strict time dependence. Furthermore, this cell death was found to be mediated both by apoptosis, which was confirmed by apoptosis-specific fluorescent staining of nuclei and cell membranes, and mitochondrial damage, as elucidated by electron microscopy. All three antioxidants used in this study partially protected against GA-induced pericyte death, suggesting that oxidative stress plays a role in GA-induced pericyte death. The results indicate that GA induces cell death in cultured bovine retinal pericytes, and that certain antioxidants may reduce this cytotoxicity.

  7. Antiviral activity of the Lippia graveolens (Mexican oregano) essential oil and its main compound carvacrol against human and animal viruses

    PubMed Central

    Pilau, Marciele Ribas; Alves, Sydney Hartz; Weiblen, Rudi; Arenhart, Sandra; Cueto, Ana Paula; Lovato, Luciane Teresinha

    2011-01-01

    Mexican oregano (Lippia graveolens) is a plant found in Mexico and Central America that is traditionally used as a medicinal herb. In the present study, we investigated the antiviral activity of the essential oil of Mexican oregano and its major component, carvacrol, against different human and animal viruses. The MTT test (3–4,5-dimethythiazol-2yl)-2,5-diphenyl tetrazolium bromide) was conducted to determine the selectivity index (SI) of the essential oil, which was equal to 13.1, 7.4, 10.8, 9.7, and 7.2 for acyclovir-resistant herpes simplex virus type 1 (ACVR-HHV-1), acyclovir-sensitive HHV-1, human respiratory syncytial virus (HRSV), bovine herpesvirus type 2 (BoHV-2), and bovine viral diarrhoea virus (BVDV), respectively. The human rotavirus (RV) and BoHV-1 and 5 were not inhibited by the essential oil. Carvacrol alone exhibited high antiviral activity against RV with a SI of 33, but it was less efficient than the oil for the other viruses. Thus, Mexican oregano oil and its main component, carvacrol, are able to inhibit different human and animal viruses in vitro. Specifically, the antiviral effects of Mexican oregano oil on ACVR-HHV-1 and HRSV and of carvacrol on RV justify more detailed studies. PMID:24031796

  8. In vitro study of RRS HA injectable mesotherapy/biorevitalization product on human skin fibroblasts and its clinical utilization

    PubMed Central

    Deglesne, Pierre-Antoine; Arroyo, Rodrigo; Ranneva, Evgeniya; Deprez, Philippe

    2016-01-01

    Mesotherapy/biorevitalization with hyaluronic acid (HA) is a treatment approach currently used for skin rejuvenation. Various products with a wide range of polycomponent formulations are available on the market. Most of these formulations contain noncross-linked HA in combination with a biorevitalization cocktail, formed by various amounts of vitamins, minerals, amino acids, nucleotides, coenzymes, and antioxidants. Although ingredients are very similar among the different products, in vitro and clinical effects may vary substantially. There is a real need for better characterization of these products in terms of their action on human skin or in vitro skin models. In this study, we analyzed the effect of the RRS® (Repairs, Refills, Stimulates) HA injectable medical device on human skin fibroblasts in vitro. Skin fibroblast viability and its capacity to induce the production of key extracellular matrix were evaluated in the presence of different concentrations of RRS HA injectable. Viability was evaluated through colorimetric MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay, and key extracellular matrix genes, type I collagen and elastin, were quantified by quantitative polymerase chain reaction. Results demonstrated that RRS HA injectable could promote human skin fibroblast viability (+15%) and increase fibroblast gene expression of type I collagen and elastin by 9.7-fold and 14-fold in vitro, respectively. These results demonstrate that mesotherapy/biorevitalization products can, at least in vitro, effectively modulate human skin fibroblasts. PMID:26966384

  9. Effect of 940 nm low-level laser therapy on osteogenesis in vitro

    NASA Astrophysics Data System (ADS)

    Jawad, Mohammed Mahmood; Husein, Adam; Azlina, Ahmad; Alam, Mohammad Khursheed; Hassan, Rozita; Shaari, Rumaizi

    2013-12-01

    Bone regeneration is essential in medical treatment, such as in surgical bone healing and orthodontics. The aim of this study is to examine the effect of different powers of 940 nm diode low-level laser treatment (LLLT) on osteoblast cells during their proliferation and differentiation stages. A human fetal osteoblast cell line was cultured and treated with LLLT. The cells were divided into experimental groups according to the power delivered and periods of exposure per day for each laser power. The (3-(4,5-dimethylthiazol-2yl)-2,5 diphenyl tetrazolium bromide) (MTT) assay was used to determine cell proliferation. Both alkaline phosphatase and osteocalcin activity assays were assessed for cell differentiation. All treatment groups showed a significant increase in cell proliferation and differentiation compared to the control group. Regarding the exposure time, the subgroups treated with the LLLT for 6 min showed higher proliferation and differentiation rates for the powers delivered, the 300-mW LLLT group significantly increased the amount of cell proliferation. By contrast, the 100 and 200 mW groups showed significantly greater amounts of cell differentiation. These results suggest that the use of LLLT may play an important role in stimulating osteoblast cells for improved bone formation.

  10. The effects of six root-end filling materials and their leachable components on cell viability.

    PubMed

    Al-Sa'eed, Oula R; Al-Hiyasat, Ahmad S; Darmani, Homa

    2008-11-01

    This study investigated the effect of six root-end filling materials (Retroplast [Retroplast Trading, Dybesøvej, Denmark], Geristore, [DEN-MAT Corporation, Santa Maria, CA], Ketac Fil [3M ESPE, Seefeld, Germany], IRM [Caulk-Dentsply, Milford, DE], Super EBA [Bosworth Company, Skokie, IL], and MTA [Dentsply-Tulsa Dental, Johnson City, TN]) on the viability of Balb/C 3T3 fibroblasts using the [3-4, 5-dimethyl thiazol-2-yl]-2,5-diphenyl tetrazolium bromide succinate (MTT) assay. Ten discs (5 mm x 2 mm) of each material were eluted in tissue culture medium for 24 hours at 37 degrees C for 3 successive days and the elutes used for cell viability testing and for determination of leached components. The results showed that Retroplast, Geristore, and Ketac Fil increased cell proliferation, whereas Super-EBA decreased cell viability. The proliferative effect of Retroplast and Geristore increased with the eluting time (24 hours, 48 hours, and 72 hours), whereas the effects of the other materials did not significantly change. IRM and MTA did not affect cell viability. High-performance liquid chromatography analysis and atomic absorption spectroscopy showed that there was variation in the amount of leached components from the materials. Our results indicate that the reaction of cells to root-end filling materials varies considerably between materials.

  11. In vitro antiproliferativeactivity of Annona reticulata roots on human cancer cell lines

    PubMed Central

    Suresh, H. M.; Shivakumar, B.; Hemalatha, K.; Heroor, S. S.; Hugar, D. S.; Rao, K. R. S. Sambasiva

    2011-01-01

    Background: The phytochemical and pharmacological activities of Annona reticulata components suggest a wide range of clinical application in lieu of cancer chemotherapy. Materials and Methods: Ethanol and aqueous extracts of roots of Annona reticulata Linn were studied for their in vitro antiproliferative activity on A-549 (human lung carcinoma), K-562 (human chronic myelogenous leukemia bone marrow), HeLa (human cervix) and MDA-MB (human adenocarcinoma mammary gland) cancer cell lines by MTT [3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide] colorimetric assay. Results: The ethanol extract exhibited a prominent inhibitory effect against A-549, K-562, HeLa and MDA-MB human cancer cell lines at a concentration range between 10 and 40 μg/ml, whereas the aqueous extract showed a lower activity at the same concentration. Simultaneously, the effect of the ethanol extract toward the inhibition of Vero cell line proliferation was lower in comparison with the cancer cell lines. Conclusion: The significant antiproliferative activity of the ethanol extract of Annona reticulata roots against A-549, K-562, HeLa and MDA-MB human cancer cell lines may be attributed toward the collective presence of acetogenins, alkaloids and lower inhibitory effect on Vero cell line, which suggests Annona reticulata be used as a chemopreventive agent in cancer therapy. PMID:21731389

  12. Black Soybean Extract Protects Against TMT-Induced Cognitive Defects in Mice

    PubMed Central

    Jeong, Ji Hee; Jo, Yu Na; Kim, Hyeon Ju; Jin, Dong Eun; Kim, Dae-Ok

    2014-01-01

    Abstract To find a neuroactive compound with a potent inhibitory effect on acetylcholinesterase (AChE) and in vivo anti-amnesic activity from natural resources, we evaluated anthocyanins and nonanthocyanins from black soybean extract. Nonanthocyanins from black soybean extract were the most potent and dose-dependent AChE inhibitors. Intracellular reactive oxygen species accumulation resulting from H2O2 treatment was significantly decreased compared with cells treated with H2O2 only. Nonanthocyanins were also neuroprotective against H2O2 treated neurotoxicity by 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assay. Finally, nonanthocyanins from black soybean in the preadministration group attenuated trimethyltin (TMT)-induced memory injury in both in vivo tests. AChE, prepared from mice brain tissues, was inhibited by nonanthocyanins from black soybean in a dose-dependent manner. Malondialdehyde generation in the brain homogenates of mice treated with nonanthocyanins from black soybean was decreased. We concluded that nonanthocyanins from black soybean had an efficacious in vitro AChE inhibitory activity, and protected against H2O2-induced neurotoxicity. In addition, our findings suggest that nonanthocyanins from black soybean may improve the TMT-induced learning and memory deficit because of AChE inhibition of mice brain tissue. Consequently, these results demonstrate that the nonanthocyanins from black soybean could possess a wide range of beneficial activities for neurodegenerative disorders. PMID:24456358

  13. Phagocytosis of PLGA Microparticles in Rat Peritoneal Exudate Cells: A Time-Dependent Study

    NASA Astrophysics Data System (ADS)

    Gomes, Anderson De Jesus; Nain Lunardi, Claure; Henrique Caetano, Flávio; Orive Lunardi, Laurelúcia; da Hora Machado, Antonio Eduardo

    2006-07-01

    With the purpose of enhancing the efficacy of microparticle-encapsulated therapeutic agents, in this study we evaluated the phagocytic ability of rat peritoneal exudate cells and the preferential location of poly(D,L-lactide-co-glycolic acid) (PLGA) microparticles inside these cells. The microparticles used were produced by a solvent evaporation method and were characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). Size distribution analysis using DLS and SEM showed that the particles were spherical, with diameters falling between 0.5 and 1.5 [mu]m. Results from cell adhesion by SEM assay, indicated that the PLGA microparticles are not toxic to cells and do not cause any distinct damage to them as confirmed by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. Among the large variety of cell populations found in the peritoneal exudates (neutrophils, eosinophils, monocytes, and macrophages), TEM showed that only the latter phagocytosed PLGA microparticles, in a time-dependent manner. The results obtained indicate that the microparticles studied show merits as possible carriers of drugs for intracellular delivery.

  14. Noscapine protects OLN-93 oligodendrocytes from ischemia-reperfusion damage: Calcium and nitric oxide involvement.

    PubMed

    Nadjafi, S; Ebrahimi, S-A; Rahbar-Roshandel, N

    2015-12-01

    This study was carried out to evaluate the effects of noscapine, a benzylisoquinoline alkaloid from opium poppy, on oligodendrocyte during ischemia/reperfusion-induced excitotoxic injury. Changes in intracellular calcium levels due to chemical ischemia and nitric oxide (NO) production during ischemia/reperfusion were evaluated as the hallmarks of ischemia-derived excitotoxic event. OLN-93 cell line (a permanent immature rat oligodendrocyte) was used as a model of oligodendrocyte. 30- or 60-minute-oxygen-glucose deprivation/24 hours reperfusion were used to induce excitotoxicity. MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay was used to evaluate cell viability. Ratiometric fluorescence microscopy using Ca(2+)-sensitive indicator Fura-2/AM was utilized to assess intracellular calcium levels. NO production was evaluated by Griess method. Noscapine (4 μM) significantly attenuated intracellular Ca(2+) elevation (P < 0.001). Also, noscapine significantly decreased NO production during a 30-minute oxygen-glucose deprivation/reperfusion (P < 0.01). The inhibitory effect of noscapine (4 μM) on intracellular Ca(2+) was greater than ionotropic glutamate receptors antagonists. Noscapine is protective against ischemia/reperfusion-induced excitotoxic injury in OLN-93 oligodendrocyte. This protective effect seems to be related to attenuation of intracellular Ca(2+) overload and NO production.

  15. Antioxidant and Anticancer Activities of Walnut (Juglans regia L.) Protein Hydrolysates Using Different Proteases.

    PubMed

    Jahanbani, Raheleh; Ghaffari, S Mahmood; Salami, Maryam; Vahdati, Kourosh; Sepehri, Houri; Sarvestani, Nazanin Namazi; Sheibani, Nader; Moosavi-Movahedi, Ali Akbar

    2016-12-01

    Walnut (Juglans regia L.) contains approximately 20-25 % protein with abundant essential amino acids. The enzymatic hydrolysate of Persian walnut (Chandler) seed proteins was prepared by incubation with three different proteases, including pancreatic chymotrypsin and trypsin, and a microbial enzyme proteinase K. The hydrolysates were found to possess excellent antioxidant capacities. The peptide fractions scavenged the 2, 2'-anizo-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) free radicals and inhibited the activity of reactive oxygen species. Walnut protein hydrolysates were also tested, for the first time, against the viability of human breast (MDA-MB231) and colon (HT-29) cancer cell lines. MTT, [3-(4, 5dimethylthiazolyl)-2,5-diphenyl-tetrazolium bromide], assay was used to assess in vitro cancer cell viability upon treatment with the peptide fractions. The peptide fractions showed cell growth inhibition of 63 ± 1.73 % for breast cancer and 51 ± 1.45 % for colon cancer cells. Thus, a direct correlation between antioxidant and anticancer activities of walnut peptide fractions exists and supports their potential therapeutic benefit.

  16. Titania-polymeric powder coatings with nano-topography support enhanced human mesenchymal cell responses.

    PubMed

    Mozumder, Mohammad Sayem; Zhu, Jesse; Perinpanayagam, Hiran

    2012-10-01

    Titanium implant osseointegration is dependent on the cellular response to surface modifications and coatings. Titania-enriched nanocomposite polymeric resin coatings were prepared through the application of advanced ultrafine powder coating technology. Their surfaces were readily modified to create nano-rough (<100 nm) surface nano-topographies that supported human embryonic palatal mesenchymal cell responses. Energy dispersive x-ray spectroscopy confirmed continuous and homogenous coatings with a similar composition and even distribution of titanium. Scanning electron microscopy (SEM) showed complex micro-topographies, and atomic force microscopy revealed intricate nanofeatures and surface roughness. Cell counts, mitochondrial enzyme activity reduction of yellow 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to dark purple, SEM, and inverted fluorescence microscopy showed a marked increase in cell attachment, spreading, proliferation, and metabolic activity on the nanostructured surfaces. Reverse Transcription- Polymerase Chain Reaction (RT-PCR) analysis showed that type I collagen and Runx2 expression were induced, and Alizarin red staining showed that mineral deposits were abundant in the cell cultures grown on nanosurfaces. This enhancement in human mesenchymal cell attachment, growth, and osteogenesis were attributed to the nanosized surface topographies, roughness, and moderate wetting characteristics of the coatings. Their dimensional similarity to naturally occurring matrix proteins and crystals, coupled with their increased surface area for protein adsorption, may have facilitated the response. Therefore, this application of ultrafine powder coating technology affords highly biocompatible surfaces that can be readily modified to accentuate the cellular response.

  17. Preparation, characterization, and anticancer efficacy of evodiamine-loaded PLGA nanoparticles.

    PubMed

    Zou, Lidi; Chen, Fengqian; Bao, Jiaolin; Wang, Shengpeng; Wang, Lu; Chen, Meiwan; He, Chengwei; Wang, Yitao

    2016-01-01

    Evodiamine (EVO) is a plant-derived indolequinazoline alkaloid with potential anticancer activity. However, low bioavailability caused by its poor water solubility limits it anticancer efficacy in clinic. To enhance the solubility and improve the bioavailability of EVO, a delivery system based on poly (lactic-co-glycolic acid) (PLGA) nanoparticles loaded with EVO (EVO-PLGA NPs) for treating breast cancer was prepared in this study. The physicochemical characterization and in vitro antitumor evaluation of EVO-PLGA NPs were determined. EVO-PLGA NPs could persistently control the release of EVO for 180 h. 3-[4,5-Dimethyl-2-thiazolyl]-2,5-diphenyl tetrazolium bromide (MTT) assessment and colony formation assay showed that EVO-PLGA NPs could enhance the toxicity and the proliferation inhibition effect of EVO on MCF-7 breast cancer cells. EVO-PLGA NPs did not strengthen G2/M arrest effect of EVO-treated cells after 24h incubation. Meanwhile, EVO-PLGA NPs could increase the expression of cyclin B1 and decrease the expression of β-actin. Taken together, these results suggested that -PLGA NPs is promising for improving anticancer efficacy of EVO in breast cancer therapy.

  18. Silica nanowire conjugated with loop-shaped oligonucleotides: A new structure to silence cysteine proteinase gene in Leishmania tropica.

    PubMed

    Bafghi, Ali Fatahi; Jebali, Ali; Daliri, Karim

    2015-12-01

    The main aim of this study was to evaluate the capability of silica nanowire conjugated with loop-shaped oligonucleotides (SNWCLSOs) to silence cysteine proteinase b (Cpb) gene in Leishmania (L) tropica. On the other hand, its toxicity on amastigotes and mouse peritoneal macrophages was evaluated by 5-diphenyl-tetrazolium bromide (MTT) assay. For control, two loop-shaped oligonucleotides (LSO) were considered. LSO1 and LSO2 were 5'-NH2-cccccaaaaaaaaaaaaaaaaaaaaaaaaaggggg-COOH-3' and LSO2: 5'-NH2-cccccttttttttttttttttttttttttttttttttttttttggggg-COOH-3', respectively. After 72 h incubation at 37 °C, AMSNW, LSO1, and LSO2 had no remarkable toxicity on L. tropica amastigote (2 × 10(5)/mL) and mouse peritoneal macrophages (2 × 10(5)/mL). In case of SNWCLSOs, they had high toxicity on L. tropica amastigote, but they had no effect on mouse peritoneal macrophages. At concentrations of 1, 10, and 25 μg/mL, AMSNW, LSO1 and LSO2 had no effect on the gene expression. But, at concentration of 50 and 100 μg/mL, decrease of gene expression was observed. In case of SNWCLSOs, they could dramatically decrease the gene expression. It could be concluded that since SNWCLSOs could silence Cpb gene with no remarkable toxicity, they are good choice for treat cutaneous leishmaniasis in future. As a new agent, it must be checked in vivo.

  19. Diethylentriaminepenta acetic acid glucose conjugates as a cell permeable iron chelator

    PubMed Central

    Mosayebnia, Mona; Shafiee-Ardestani, Mehdi; Pasalar, Parvin; Mashayekhi, Mojgan; Amanlou, Massoud

    2014-01-01

    Objective: To find out whether DTPA-DG complex can enhance clearance of intracellular free iron. Materials and Methods: Diethylenetriaminepentaacetic acid-D-deoxy-glucosamine (DTPA-DG) was synthesized and examined for its activity as a cell-permeable iron chelator in human hepatocellular carcinoma (HEPG2) cell line exposed to high concentration of iron sulfate and compared with deferoxamine (DFO), a prototype iron chelator. The effect of DTPA-DG on cell viability was monitored using the 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide MTT assay as well. Results: There was a significant increase of iron level after iron overload induction in HEPG2 cell culture. DTPA-DG presented a remarkable capacity to iron burden reducing with estimated 50% inhibitory concentration value of 65.77 nM. In fact, glycosyl moiety was gained access of DTPA to intracellular iron deposits through glucose transporter systems. Conclusion: DTPA-DG, more potent than DFO to sequester deposits of free iron with no profound toxic effect. The results suggest the potential of DTPA-DG in chelating iron and permitting its excretion from primary organ storage. PMID:24554907

  20. Potential protection of green tea polyphenols against 1800 MHz electromagnetic radiation-induced injury on rat cortical neurons.

    PubMed

    Liu, Mei-Li; Wen, Jian-Qiang; Fan, Yu-Bo

    2011-10-01

    Radiofrequency electromagnetic fields (EMF) are harmful to public health, but the certain anti-irradiation mechanism is not clear yet. The present study was performed to investigate the possible protective effects of green tea polyphenols against electromagnetic radiation-induced injury in the cultured rat cortical neurons. In this study, green tea polyphenols were used in the cultured cortical neurons exposed to 1800 MHz EMFs by the mobile phone. We found that the mobile phone irradiation for 24 h induced marked neuronal cell death in the MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazolium bromide) and TUNEL (TdT mediated biotin-dUTP nicked-end labeling) assay, and protective effects of green tea polyphenols on the injured cortical neurons were demonstrated by testing the content of Bcl-2 Assaciated X protein (Bax) in the immunoprecipitation assay and Western blot assay. In our study results, the mobile phone irradiation-induced increases in the content of active Bax were inhibited significantly by green tea polyphenols, while the contents of total Bax had no marked changes after the treatment of green tea polyphenols. Our results suggested a neuroprotective effect of green tea polyphenols against the mobile phone irradiation-induced injury on the cultured rat cortical neurons.

  1. Cytotoxic effect of microbial biosurfactants against human embryonic kidney cancerous cell: HEK-293 and their possible role in apoptosis.

    PubMed

    Pradhan, Arun Kumar; Pradhan, Nilotpala; Mohapatra, Purusottam; Kundu, Chanakya Nath; Panda, Prasanna Kumar; Mishra, Barada Kanta

    2014-11-01

    Two different microbial biosurfactants S9BS and CHBS were isolated from Lysinibacillus fusiformis S9 and Bacillus tequilensis CH. Cytotoxicity effect of these biosurfactants on human embryonic kidney cancerous cell (HEK-293) were studied with the help of 3-(4,5-dimethylthiazol-2yl-)-2, 5-diphenyl tetrazolium bromide (MTT) assay and morphological changes were observed under inverted microscope. The biosurfactants exhibited positive cytotoxic effect on HEK-293 cell line. It was found that LC50 of S9BS and CHBS were 75 and 100 μg ml(-1), respectively. Further cell cycle and apoptosis analysis of biosurfactant-treated HEK-293 cell line were done by FACS. In this study, cytotoxic effect of glycolipid biosurfactant against HEK-293 cell lines is reported for the first time. Mechanism towards increased membrane permeability of biosurfactant-treated cancer cell may be the incorporation of its lipid moiety into the plasma membrane leading to formation of pores and membrane disruption. Hence, these microbial biosurfactants can prove to be significant biomolecule for cancer treatment.

  2. Rutin inhibits proliferation, attenuates superoxide production and decreases adhesion and migration of human cancerous cells.

    PubMed

    Ben Sghaier, Mohamed; Pagano, Alessandra; Mousslim, Mohamed; Ammari, Youssef; Kovacic, Hervé; Luis, José

    2016-12-01

    Lung and colorectal cancer are the principal causes of death in the world. Rutin, an active flavonoid compound, is known for possessing a wide range of biological activities. In this study, we examined the effect of rutin on the viability, superoxide anion production, adhesion and migration of human lung (A549) and colon (HT29 and Caco-2) cancer cell lines. In order to control the harmlessness of the tested concentrations of rutin, the viability of cancer cell lines was assessed using a 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. ROS generation was measured by lucigenin chemiluminescence detecting superoxide ions. To investigate the effect of rutin on the behavior of human lung and colon cancer cell lines, we performed adhesion assays, using various purified extracellular matrix (ECM) proteins. Finally, in vitro cell migration assays were explored using modified Boyden chambers. The viability of cancerous cells was inhibited by rutin. It also significantly attenuated the superoxide production in HT29 cells. In addition, rutin affected adhesion and migration of A549 and HT29 cell. These findings indicate that rutin, a natural molecule, might have potential as anticancer agent against lung and colorectal carcinogenesis.

  3. Preparation, characterization and biocompatibility studies on risperidone-loaded solid lipid nanoparticles (SLN): high pressure homogenization versus ultrasound.

    PubMed

    Silva, A C; González-Mira, E; García, M L; Egea, M A; Fonseca, J; Silva, R; Santos, D; Souto, E B; Ferreira, D

    2011-08-01

    The suitability of solid lipid nanoparticles (SLN) for the encapsulation of risperidone (RISP), an antipsychotic lipophilic drug, was assessed for oral administration. The hot high pressure homogenization (HPH) and the ultrasound (US) technique were used as production methods for SLN. All the studies on the SLN formulations were done in parallel, in order to compare the results and conclude about the advantages and limitations of both techniques. The particle sizes were in the nanometer range for all prepared SLN formulations and the zeta potential absolute values were high, predicting good long-term stability. Optical analyses demonstrated the achievement of stable colloidal dispersions. Physicochemical characterization of dispersions and bulk lipids, performed by differential scanning calorimetry (DSC) and X-ray assays, support prediction of occurrence of drug incorporation in the SLN and good long term stability of the systems. The toxicity of SLN with Caco-2 cells and the existence of contaminations derived from the production equipments were assessed by the (4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide (MTT) assay. The results showed 90% of cell viability after SLN exposure, with no significant differences within all prepared formulations (p > 0.05). From this study, we conclude that SLN can be considered as efficient carriers for RISP encapsulation. Moreover, HPH and US revealed to be both effective methods for SLN production.

  4. Deposition of superparamagnetic nanohydroxyapatite on iron-fibrin substrates: preparation, characterization, cytocompatibility and bioactivity studies.

    PubMed

    Vedakumari, Weslen S; Priya, Vishnu M; Sastry, Thotapalli P

    2014-08-01

    In the present study, nanosized hydroxyapatite (nHAp) was formed on iron-fibrin substrates and its physico-chemical properties were characterized. The prepared iron-fibrin-nanohydroxyapatite (IF-nHAp) composite was needle shaped with an average width of about 30nm and length of 80nm. The vibrating sample magnetometer (VSM) was used to evaluate the superparamagnetic behavior of the nanocomposite, IF-nHAp. Hemolysis and ELISA (enzyme linked immunosorbent assay) were performed to evaluate the its bio/immunocompatibility and MTT (3-(4,5-dimethylazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assay using osteoblast cells was performed to scrutinize its proliferative potential. Alkaline phosphatase activity (ALP) and calcium deposition were studied to investigate the osteogenic property of the nanocomposite. RT-PCR (real time-polymerase chain reaction) was used to quantify the mRNA levels of ALP, OC (osteocalcin), and OPN (osteopontin) genes involved in the osteogenic differentiation and matrix mineralization. Further, the bone bonding ability of IF-nHAp was observed by the deposition of apatite layers on the composite incubated in simulated body fluid (SBF).

  5. Overcoming of P-glycoprotein-mediated multidrug resistance in K562/A02 cells using riccardin F and pakyonol, bisbibenzyl derivatives from liverworts.

    PubMed

    Ji, Mei; Shi, Yanquiu; Lou, Hongxiang

    2011-01-01

    Riccardin F and pakyonol, macrocyclic bisbibenzyls from Plagiochasm intermedium, have been confirmed to possess antifungic activities against Candida albicans. Herein, we evaluated their anti-tumor activity in vitro by employing K562 and K562/A02 cells, the well-known adriamycin (ADR)-induced multidrug resistance (MDR) tumor cell lines over-expressing P-glycoprotein (P-gp). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assays showed that riccardin F and pakyonol ranging from 0 to 6 μg/mL exhibited no inhibitory effects on the growth of the two cell lines. However, in the presence of 3 μg/mL riccardin F or pakyonol (non-cytotoxic concentration), the IC50 of ADR against K562/A02 cells decreased by 2.51- and 4.78-fold, respectively. Flow cytometry showed that riccardin F and pakyonol significantly enhanced the accumulation of ADR in K562/A02 cells. Furthermore, fluorescence intensity detection revealed that the two natural products remarkably increased the retention of rhodamine-123 in K562/A02 cells rather than in K562 cells, indicating that the major cause for riccardin F and pakyonol to reverse P-gp-mediated MDR in K562/A02 cells is probably due to the constrained transport activity of P-gp. This study explores the potential application of bisbibenzyl type compounds as modulators of P-gp-mediated MDR in tumor cells.

  6. Cytotoxicity evaluation of three pairs of hexabromocyclododecane (HBCD) enantiomers on Hep G2 cell.

    PubMed

    Zhang, Xiaoling; Yang, Fangxing; Xu, Chao; Liu, Weiping; Wen, Sheng; Xu, Ying

    2008-09-01

    Hexabromocyclododecanes (HBCDs) are additive brominated flame retardants mainly used in plastics and textiles. At the present time, these compounds are found in almost all environmental and human samples. In order to evaluate the environmental safety and health risk of HBCDs, the enantiomerically pure alpha-, beta-, and gamma-HBCD were prepared using high performance liquid chromatography (HPLC) on a PM-beta-CD column and the cytotoxicities of their enantiomers were evaluated in Hep G2 cells. Results from the 3-(4,5-dimethylthioazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), resazurin reduction and lactate dehydrogenase (LDH) release assays showed a good agreement that the order of cytotoxicity was gamma-HBCDbeta-HBCD>alpha-HBCD, and that significantly lower cell viability and higher LDH release were observed in all (+)-enantiomers ((+) alpha-, (+) beta- and (+) gamma-HBCD) than the corresponding (-)-forms ((-) alpha-, (-) beta- and (-) gamma-HBCD). Additionally, the formation of reactive oxygen species (ROS) induced by these HBCD enantiomers were detected. The positive correlation between the LDH release and ROS formation demonstrated that the toxic mechanism might be mediated by oxidative damage. These results suggest that environmental and human health risks of HBCDs must be evaluated at the level of individual enantiomers.

  7. γ-terpineol inhibits cell growth and induces apoptosis in human liver cancer BEL-7402 cells in vitro.

    PubMed

    Wu, Zi-Li; Yin, Zhong-Qiong; Du, Yong-Hua; Feng, Rui-Zhang; Ye, Kui-Chuan; Wei, Qin; Hu, Yong; He, Lin; Liao, Lin; Wang, Yu

    2014-01-01

    To investigate the effect of γ-terpineol on cell proliferation and apoptosis of human hepatoma BEL-7402 cells to elucidate its molecular mechanism. Here, BEL-7402 cells were treated with various concentrations (40, 80, 160, 320 and 640 μg/ml) of γ-terpineol for 48 h, cell proliferation was determined by 3-(4,5-dimethyl-thiazolyl-2)-2,5-diphenyl tetrazolium bromides (MTT) assay. Cell colony inhibition was determined by soft agar assay. Apoptosis and possible molecular mechanisms were evaluated by morphological observation, flow cytometry analysis, and DNA fragmentation assay. The γ-terpineol significantly suppressed BEL-7402 cell proliferation in a dose-dependent manner. Characteristic morphological and biochemical changes associated with apoptosis such as cells shrinkage, deformation and vacuolization of mitochondria, nuclear chromatin condensation and fragmentation, formation of apoptotic bodies were observed after BEL-7402 cells treated with γ-terpineol for 24 h and 48 h. Cell cycle were displayed by flow cytometry analysis, the γ-terpineol treatment resulted in accumulation of cells at G₁ or S phase and a blockade of cell proliferation compared to control group. Treating BEL-7402 cells with 320 μg/ml of γ-terpineol for 36 h and 48 h, a typical apoptotic "DNA ladder" was observed using DNA fragmentation assay. The present study demonstrated that possible anti-cancer mechanism of γ-terpineol on human hematomas cells is through inducing cell apoptosis to suppress tumor cell growth.

  8. Exploring Bioactive Properties of Marine Cyanobacteria Isolated from the Portuguese Coast: High Potential as a Source of Anticancer Compounds

    PubMed Central

    Costa, Margarida; Garcia, Mónica; Costa-Rodrigues, João; Costa, Maria Sofia; Ribeiro, Maria João; Fernandes, Maria Helena; Barros, Piedade; Barreiro, Aldo; Vasconcelos, Vitor; Martins, Rosário

    2013-01-01

    The oceans remain a major source of natural compounds with potential in pharmacology. In particular, during the last few decades, marine cyanobacteria have been in focus as producers of interesting bioactive compounds, especially for the treatment of cancer. In this study, the anticancer potential of extracts from twenty eight marine cyanobacteria strains, belonging to the underexplored picoplanktonic genera, Cyanobium, Synechocystis and Synechococcus, and the filamentous genera, Nodosilinea, Leptolyngbya, Pseudanabaena and Romeria, were assessed in eight human tumor cell lines. First, a crude extract was obtained by dichloromethane:methanol extraction, and from it, three fractions were separated in a Si column chromatography. The crude extract and fractions were tested in eight human cancer cell lines for cell viability/toxicity, accessed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and lactic dehydrogenase release (LDH) assays. Eight point nine percent of the strains revealed strong cytotoxicity; 17.8% showed moderate cytotoxicity, and 14.3% assays showed low toxicity. The results obtained revealed that the studied genera of marine cyanobacteria are a promising source of novel compounds with potential anticancer activity and highlight the interest in also exploring the smaller filamentous and picoplanktonic genera of cyanobacteria. PMID:24384871

  9. Antibacterial nanosilver coated orthodontic bands with potential implications in dentistry

    PubMed Central

    Prabha, Rahul Damodaran; Kandasamy, Rajasigamani; Sivaraman, U. Sajeev; Nandkumar, Maya A.; Nair, Prabha D.

    2016-01-01

    Background & objectives: Fixed orthodontic treatment, an indispensable procedure in orthodontics, necessitates insertion of dental bands. Insertion of band material could also introduce a site of plaque retention. It was hypothesized that band materials with slow-release antimicrobial properties could help in sustained infection control, prevention of dental plaque formation and further associated health risks. Considering the known antimicrobial proprieties of silver, a coating of silver nanoparticle (SNP) onto the stainless steel bands was done and characterized for its beneficial properties in the prevention of plaque accumulation. Methods: Coatings of SNPs on conventional stainless steel dental bands were prepared using thermal evaporation technology. The coated dental bands were characterized for their physicochemical properties and evaluated for antimicrobial activity and biocompatibility. The physiochemical characterization of band material both coated and uncoated was carried out using scanning electron microscope, energy dispersive spectroscopy, atomic force microscopyand contact angle test. Biocompatibility tests for coated band material were carried using L929 mouse fibroblast cell culture and MTT [3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide] assay. Antimicrobial activity of coated band material against Gram-positive bacteria was tested. Results: A stable and uniform coating of SNPs was obtained. The coated band materials were biocompatible as well as possessed distinct antimicrobial activity. Interpretation & conclusions: The SNP coated dental bands could be potential antimicrobial dental bands for future clinical use. Further studies need to be done to validate the efficiency of coated band materials in oral environments. PMID:28256467

  10. MTT assay for cell viability: Intracellular localization of the formazan product is in lipid droplets.

    PubMed

    Stockert, Juan C; Blázquez-Castro, Alfonso; Cañete, Magdalena; Horobin, Richard W; Villanueva, Angeles

    2012-12-01

    Although MTT is widely used to assess cytotoxicity and cell viability, the precise localization of its reduced formazan product is still unclear. In the present study the localization of MTT formazan was studied by direct microscopic observation of living HeLa cells and by colocalization analysis with organelle-selective fluorescent probes. MTT formazan granules did not colocalize with mitochondria as revealed by rhodamine 123 labeling or autofluorescence. Likewise, no colocalization was observed between MTT formazan granules and lysosomes labeled by neutral red. Taking into account the lipophilic character and lipid solubility of MTT formazan, an evaluation of the MTT reaction was performed after treatment of cells with sunflower oil emulsions to induce a massive occurrence of lipid droplets. Under this condition, lipid droplets revealed a large amount of MTT formazan deposits. Kinetic studies on the viability of MTT-treated cells showed no harmful effects at short times. Quantitative structure-activity relations (QSAR) models were used to predict and explain the localization of both the MTT tetrazolium salt and its formazan product. These predictions were in agreement with experimental observations on the accumulation of MTT formazan product in lipid droplets.

  11. Mitomycin C-induced cell death in mouse lens epithelial cells.

    PubMed

    Park, Hyun-Kyung; Lee, Kwang-Won; Choi, Jun-Sub; Joo, Choun-Ki

    2002-01-01

    The mode of action of mitomycin C (MMC)-induced cell death in lens epithelia was investigated using cell culture system. The cytotoxicity of MMC on alpha-TN4 mouse lens epithelial cells (LECs) was measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT)-based assay. To determine the type of cell death induced by MMC, we stained LECs with acridine orange and ethidium bromide, and performed TUNEL assay and electron microscopic examination. The activation of MMC was also investigated with N-acetly-L-cystein (NAC), a free radical scavenger, and dicumarol, an inhibitor of DT-diaphorase, to analyze one- and two-electron reduction pathways involved in the activation of MMC. MTT assay showed that 400 microg/ml of MMC decreased the cell viability of the control cells to 45%, and the cytotoxicity of MMC on alpha-TN4 cells was increased in a dose-dependent manner. Based on the results obtained from acridine orange and ethidium bromide staining, the TUNEL assay, and transmission electron microscopic examination, we confirmed that MMC-induced cell death, at 400 microg/ml of MMC, occurs with necrosis as well as apoptotis. The treatment of cells with 2 mM NAC for 1 h or 20 microM dicumarol for 30 min, prior to 5 min of MMC (400 microg/ml) treatment, restored the growth suppression to 76 and 80% of the control level, respectively. In addition, the cotreatment of cells with NAC and dicumarol restored cell viability to 90% of the control level. The cellular death in MMC-treated LECs showed the involvement of oxidative stress in this apoptosis accounting for 22% of the observed cell death at 400 microg/ml of MMC.

  12. [Determination of Escherichia coli Shiga-like toxins by means of the MTT bioassay].

    PubMed

    Hörmansdorfer, S; Gareis, M; Bauer, J; Mayr, A

    1995-09-01

    Tissue culture cells' metabolism and viability are measured by the mitochondrial reduction rate of a yellow tetrazolium salt (MTT) to blue formazan crystals in the MTT-bioassay. Thus the MTT-bioassay is a standardizable and reproducible bioassay for measuring cytotoxicity or cytostimulation. It is shown that the MTT-bioassay is also very suitable for determining bacterial cytotoxins using Escherichia coli's Shiga-like toxins as example. 177 strains of E. coli, isolated from carcasses and organs of cattle, are classified biochemically and tested for cytotoxin production by means of the MTT-bioassay. One of these strains is recognized as producer of Shiga-like toxin 2. 4 Enterohemolysin-producing strains of E. coli are cultivated from a feces sample of a diarrhoeic nubian ibex and identified as Shiga-like toxin 1 producers by help of the MTT-bioassay.

  13. Cholesterol does not affect the toxicity of amyloid beta fragment but mimics its effect on MTT formazan exocytosis in cultured rat hippocampal neurons.

    PubMed

    Abe, K; Saito, H

    1999-12-01

    It has recently been reported that methyl-beta-cyclodextrin-solubilized cholesterol protects PC12 cells from amyloid beta protein (Abeta) toxicity. To ask if this is the case in brain neurons, we investigated its effect in primary cultured rat hippocampal neurons. In basal culture conditions with no addition of Abeta, methyl-beta-cyclodextrin-solubilized cholesterol at concentrations of 30-100 microM was toxic to neurons, but at concentrations of 1-10 microM promoted neuronal survival. Methyl-beta-cyclodextrin-solubilized cholesterol at 1-10 microM was also effective in protecting neurons from toxicity of 20 microM Abeta. However, these effects were all mimicked by methyl-beta-cyclodextrin alone, but not by cholesterol solubilized by dimethylsulfoxide or ethanol. The effects of methyl-beta-cyclodextrin-solubilized cholesterol on neuronal survival and Abeta toxicity are probably attributed to the action of methyl-beta-cyclodextrin, but not cholesterol. Alternatively, we found that methyl-beta-cyclodextrin-solubilized cholesterol at lower concentrations ( > 10 nM) inhibited cellular reduction of 3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) by promoting the exocytosis of MTT formazan. This effect was shared by dimethylsulfoxide- or ethanol-solubilized cholesterol, but not by methyl-beta-cyclodextrin, supporting that it is attributed to the action of cholesterol. These results suggest that cholesterol does not protect neurons from Abeta toxicity, or rather inhibits cellular MTT reduction in a similar manner to Abeta.

  14. Biocompatibility evaluation of cigarette and carbon papers used in repair of traumatic tympanic membrane perforations: experimental study.

    PubMed

    Altuntaş, Emine Elif; Sümer, Zeynep

    2013-01-01

    The purposes of this study were to investigate the biocompatibility of two different paper patches (carbon and cigarette papers) and compare the adhesion and proliferation features of L929 fibroblast cells by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT Test) test and scanning electron microscopy (SEM). In this study, time-dependent cytotoxic effects of cigarette and carbon papers used in repairing small traumatic TM perforations were investigated in vitro by using MTT test. And also adhesion and spreading of cells over disk surface were observed by SEM. Cytotoxicity test carried out by MTT analysis on leakage products collected from two types of paper patches at the end of 24 and 48 h revealed no cytotoxicity (P > 0.05). In SEM studies, it was observed that cells started to proliferate over disk surface as a result of 48-h incubation, and SEM revealed that the cell proliferation over cigarette paper was more compared to the one over carbon paper. We believe that this is the first study where biocompatibility and adhesion features of carbon and cigarette paper have been studied by using L929 fibroblast cell culture. As a result, biocompatibility of cigarette paper and also whether cigarette paper was superior to carbon paper in cell attachment and biocompatibility were studied. It was found, by MTT test and SEM test, that cigarette paper had a higher biocompatibility and cell attachment, and thus cigarette paper should be the patch to be preferred in cases where TM perforations are repaired by paper-patch method.

  15. Protective effect of zinc chloride against cobalt chloride-induced cytotoxicity on vero cells: preliminary results.

    PubMed

    Gürbay, Aylin

    2012-07-01

    The aim of this study was to investigate the possible time- and dose-dependent cytotoxic effects of cobalt chloride on Vero cells. The cultured cells were incubated with different concentrations of cobalt chloride ranging from 0.5 to 1,000 μM, and cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) and resazurin assays. Possible protective effects of vitamin E, coenzyme Q(10), and zinc chloride were also tested in this system. A gradual decrease in cell proliferation was observed at concentrations ~≥ 200 μM in incubation periods of 24, 48, 72, and 96 h with MTT assay. Exposure of cells to 500 and 1,000 μM cobalt chloride caused significant decrease in cell survival. A biphasic survival profile of cells was observed at 1-25 μM concentration range following 96 h of incubation. With resazurin assay, cytotoxicity profile of CoCl(2) was found comparable to the results of MTT assay, particularly at high concentrations and long incubation periods. Dose-dependent cytotoxicity was noted following exposure of cells to ≥ 250 μM of CoCl(2) for 24 h and ≥ 100 μM concentrations of CoCl(2) for 48-96 h. Pretreatment of cells with ZnCl(2) for 4 or 24 h provided significant protection against cobalt chloride-induced cytotoxicity when measured with MTT assay. However, vitamin E or coenzyme Q(10) was not protective. CoCl(2) had dose- and time-dependent cytotoxic effects in Vero cells. Preventive effect of ZnCl(2) against CoCl(2)-induced cytotoxicity should be considered in detail to define exact mechanism of toxicity in Vero cells.

  16. The effects of biocides (antiseptics and disinfectants) on the endospores of Rhinosporidium seeberi.

    PubMed

    Arseculeratne, S N; Atapattu, D N; Balasooriya, P; Fernando, R

    2006-04-01

    No data exists on the activity of biocides (antiseptics and disinfectants) on Rhinosporidium seeberi that causes rhinosporidiosis in humans and animals. On account of the inability to culture R. seeberi, in vitro, dyes were used to assess the morphological integrity and viability of biocide-treated endospores that are considered to be the infective stage of this pathogen. Evan's Blue (EvB) identifies the morphological integrity of the endospores while MTT (3-[4, 5-dimethylthiazol-2yl]-2, 5-diphenyl tetrazolium bromide) identifies metabolic activity through its reduction by cellular dehydrogenases to microscopically visible deposits of insoluble formazan. MTT-negativity has earlier been shown to correlate with absence of growth of yeast and mycelial fungi in culture and could thus indicate the loss of viability of MTT-negative rhinosporidial endospores. Hydrogen peroxide, glutaraldehyde, chloroxylenol, chlorhexidine, cetrimide, thimerosal, 70% ethanol, iodine in 70% ethanol, 10% formalin, povidone-iodine, sodium azide and silver nitrate were tested on freshly-harvested endospores and all biocides caused metabolic inactivation with or without altered structural integrity as shown by absence of MTT-staining after 3, 24 or 36 hour after exposure, while EvB stained only the endospores treated with sodium azide, ethanol, thimerosal, chloroxylenol, glutaraldehyde and hydrogen peroxide. With clinically useful biocides - chlorhexidine, cetrimide-chlorhexidine, 70% ethanol, povidone-iodine and silver nitrate, a total period of exposure of endospores to the biocide, for seven minutes, produced metabolic inactivation of the endospores. Anti-rhinosporidial antiseptics that could be used in surgery on rhinosporidial patients include povidone-iodine in nasal packs for nasal and naso-pharyngeal surgery, chlorhexidine and cetrimide-chlorhexidine on the skin, while povidone-iodine and silver nitrate could have application in ocular rhinosporidiosis.

  17. Bromide Adsorption by Reference Minerals and Soils

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bromide, Br-, adsorption behavior was investigated on amorphous Al and Fe oxide, montmorillonite, kaolinite, and temperate and tropical soils. Bromide adsorption decreased with increasing solution pH with minimal adsorption occurring above pH 7. Bromide adsorption was higher for amorphous oxides t...

  18. Nickel nanowires induced and reactive oxygen species mediated apoptosis in human pancreatic adenocarcinoma cells

    PubMed Central

    Hossain, Md. Zakir; Kleve, Maurice G

    2011-01-01

    Background The ability to evade apoptosis is one of the key properties of cancer. The apoptogenic effect of nickel nanowires (Ni NWs) on cancer cell lines has never been adequately addressed. Due to the unique physicochemical characteristics of Ni NWs, we envision the development of a novel anticancer therapeutics specifically for pancreatic cancer. Thus, we investigated whether Ni NWs induce ROS-mediated apoptosis in human pancreatic adenocarcinoma (Panc-1) cells. Methods In this study Ni NWs were fabricated using the electrodeposition method. Synthesized Ni NWs were physically characterized by energy dispersive X-ray analysis, UV-Vis spectroscopy of NanoDrop 2000 (UV-Vis), magnetization study, scanning electron microscopy, and transmission electron microscopy. Assessment of morphological apoptotic characteristics by phase contrast microscopy (PCM), Ni-NWs-induced apoptosis staining with ethidium bromide (EB) and acridine orange (AO) followed by fluorescence microscopy (FM) was performed. For molecular biological and biochemical characterization, Panc-1 cell culture and cytotoxic effect of Ni NWs were determined by using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Quantitative apoptosis was analyzed by flow cytometry staining with propidium iodide through cell cycle arrest and generation of ROS using 2′, 7′-dichlorofluorescein diacetate fluorescence intensity. In all experiments, Panc-1 cancer cells without any treatment were used as the negative controls. Results The intracellular uptake of Ni NWs through endocytosis by Panc-1 cells was observed by PCM. EB and AO staining of FM and MTT assay qualitatively and quantitatively confirmed the extent of apoptosis. Flow cytometric cell cycle arrest and ROS generation indicated Ni NWs as inducers of apoptotic cell death. Conclusion We investigated the role of Ni NWs as inducers of ROS-mediated apoptosis in Panc-1 cells. These results suggested that Ni NWs could be an effective

  19. Trastuzumab and docetaxel in a preclinical organotypic breast cancer model using tissue slices from mammary fat pad: Translational relevance.

    PubMed

    Vesci, Loredana; Carollo, Valeria; Roscilli, Giuseppe; Aurisicchio, Luigi; Ferrara, Fabiana Fosca; Spagnoli, Luigi; De Santis, Rita

    2015-09-01

    With the ever-increasing number of drugs approved to treat cancers, selection of the optimal treatment regimen for an individual patient is challenging. Breast cancer complexity requires novel predictive methods and tools. In the present study, we set up experimental conditions to obtain an 'ex vivo' organotypic culture from xenotransplanted mice aiming at recapitulating the human clinical condition. The effect of trastuzumab (large biological molecule) and docetaxel (small chemical entity) was subsequently investigated on this organotypic model and compared with in vivo and in vitro activity on tumor cells. Tissue slices of 200 µm were obtained from mammary fat pad of SCID mice xenotransplanted with human MCF-7 breast cancer cells. Viability and proliferation were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) colorimetric assay and Ki-67 immunohistochemistry,and apoptosis by cleaved caspase-3 immunohistochemistry. In vivo antitumor activity of trastuzumab and docetaxel was determined by caliper measurement of tumor volume and Ki-67 expression on explanted masses by immunohistochemistry. A Teflon support and normoxia were necessary experimental conditions to obtain high viability of excised breast cancer infiltrated mammary fat pad slices upon 48 h cultivation, as shown by MTT proliferation assay, and Ki-67 expression. Breast cancer tissue slices treated for 48 h with trastuzumab or docetaxel showed a significant dose‑dependent reduction of viability by MTT assay. Consistently, both drugs down-modulated Ki-67 and increased cleaved caspase-3. Tumor masses collected from docetaxel- or trastuzumab‑treated mice showed a similar reduction of proliferation markers. By contrast, MCF-7 cell cultures were significantly inhibited by docetaxel but not by trastuzumab. Tumor tissue slices represent a more predictive experimental cancer model compared to cell cultures for both small and large molecule antitumor efficacy. This

  20. Kinetics of MTT-formazan exocytosis in phagocytic and non-phagocytic cells.

    PubMed

    Molinari, Beatriz L; Tasat, Deborah R; Palmieri, Mónica A; Cabrini, Rómulo L

    2005-01-01

    MTT is taken up by cells by endocytosis and reduced to formazan in the endosomal/lysosomal compartment. Formazan is deposited intracellularly as blue granules and is later exocytosed as needle-like formazan crystals. The present study involves an analysis of the pattern of exocytosis of MTT in different cell types showing clearcut differences in the response that can be associated to their ability to phagocytose. To further assess the characteristics of the exocytic mechanism of MTT/formazan, different experimental conditions were assayed. When culture medium with decreasing serum concentration was used as a metabolic modulator no variations were observed in the proportion of cells with formazan crystals. Conversely, the markedly sensitivity of phagocytic cells to increasing concentrations of genistein constituted a remarkable difference with non-phagocytic cells. These results must be considered when the modulation of MTT exocytosis is used as a signal of the progress of human diseases.

  1. Mesoporous silica nanoparticles enhance MTT formazan exocytosis in HeLa cells and astrocytes.

    PubMed

    Fisichella, Matthieu; Dabboue, Hinda; Bhattacharyya, Sanjib; Saboungi, Marie-Louise; Salvetat, Jean-Paul; Hevor, Tobias; Guerin, Martine

    2009-06-01

    We report on the observation that mesoporous silica nanoparticles (MSNs), after being endocytosed, interfere with the MTT test in HeLa cells and astrocytes by accelerating the exocytosis of formazan crystals. The stimulation of MTT formazan exocytosis is probably related to perturbation of intracellular vesicle trafficking by MSN uptake as revealed by experiments in presence of chloroquine and genistein. Similar effect has been previously observed with a number of chemicals, especially with neurotoxic beta amyloid peptides, but not with nanoparticles. We showed also that MTT reduction test gives an overestimation of the cytotoxicity of mesoporous silica nanoparticles compared to other tests such as LDH activity, WST-1 test and flow cytometry. These findings show that MTT assay should not be used for the study of MSN toxicity, and that perturbation of intracellular trafficking has to be taken into account in evaluating biocompatibility of MSNs.

  2. The Unreliability of MTT Assay in the Cytotoxic Test of Primary Cultured Glioblastoma Cells.

    PubMed

    Jo, Hwa Yeon; Kim, Yona; Park, Hyung Woo; Moon, Hyo Eun; Bae, Seongtae; Kim, JinWook; Kim, Dong Gyu; Paek, Sun Ha

    2015-09-01

    MTT assay is commonly used to assess the cellular cytotoxicity caused by anticancer drugs in glioblastomas. However, there have been some reports insisting that MTT assay exhibited non-specific intracellular reduction of tetrazolium which led to underestimated results of cytotoxicity. Here, we examine whether or not MTT assay can lead to incorrect information regarding alcohol-induced cytotoxicity on immortalized and primary glioblastoma cells. MTT assay was applied to assess the ethanol-induced cytotoxicity at various ethanol concentrations. The cellular cytotoxicity induced by different doses of ethanol was analyzed and compared through several cytotoxic assays. Ethanol-induced cytotoxicity observed through MTT assay on both cell types was shown to be ethanol dose-dependent below a 3% concentration. However, the cytotoxicity was shown to be markedly underestimated only in primary cells at a 5% concentration. RT-PCR and Western Blot showed increased expressions of pro-apoptotic proteins and decreased expressions of anti-apoptotic proteins in an ethanol dose-dependent manner in both cell types. Furthermore, we present a possible mechanism for the unreliable result of MTT assay. A high concentration of ethanol induces more severe membrane damage and increased intracellular concentration of NADH in primary cells which enhances the nonspecific reduction of tetrazolium salt. Together, our findings demonstrate that the cytotoxicity on primary cells could inaccurately be assessed when detected through MTT assay. Therefore, a careful interpretation is needed when one would analyze the cytotoxic results of MTT assay, and it is suggested that other assays must be accompanied to produce more reliable and accurate cytotoxic results on primary glioblastoma cells.

  3. Lanthanum Bromide Detectors for Safeguards Measurements

    SciTech Connect

    Wright, J.

    2011-05-25

    Lanthanum bromide has advantages over other popular inorganic scintillator detectors. Lanthanum bromide offers superior resolution, and good efficiency when compared to sodium iodide and lanthanum chloride. It is a good alternative to high purity germanium detectors for some safeguards applications. This paper offers an initial look at lanthanum bromide detectors. Resolution of lanthanum bromide will be compared lanthanum chloride and sodium-iodide detectors through check source measurements. Relative efficiency and angular dependence will be looked at. Nuclear material spectra, to include plutonium and highly enriched uranium, will be compared between detector types.

  4. Surface and volume properties of dodecylethyldimethylammonium bromide and benzyldimethyldodecylammonium bromide I. Surface properties of dodecylethyldimethylammonium bromide and benzyldimethyldodecylammonium bromide.

    PubMed

    Harkot, Joanna; Jańczuk, Bronisław

    2009-03-15

    Surface tension measurements were carried out for aqueous solutions of two cationic surfactants: dodecylethyldimethylammonium bromide (C(12)(EDMAB)) and benzyldimethyldodecylammonium bromide (BDDAB). Isotherms and thermodynamic adsorption parameters were determined from the surface tension data. Firstly, the surface excess concentration in the adsorbed monolayer and the total concentration of the surfactants were determined, then the standard free energy of adsorption was calculated by different methods. In the calculations, different orientations of the surfactants at the adsorbed monolayer were also taken into account. From the experimental and calculated data it results that the difference in the structure of the two cationic surfactants by changing the methyl group for aryl one in their heads causes an increase of the efficiency and a decrease of the effectiveness of adsorption at water-air interface, and that the standard free energy of adsorption can be predicted from the surface tension of the surfactants assuming the aryl group to be equivalent to 3.5 methylene groups. The experimentally obtained difference between the standard free energy of adsorption of the C(12)(EDMAB) and BDDAB was in good agreement with that theoretically accounted, corresponding to the standard free energy of adsorption of the aryl group. However, the best correlation between the values was obtained when a parallel orientation of the surfactant molecules at the adsorbed monolayer was taken into account.

  5. Minocycline recovers MTT-formazan exocytosis impaired by amyloid beta peptide.

    PubMed

    Kreutzmann, Peter; Wolf, Gerald; Kupsch, Kathleen

    2010-10-01

    Minocycline, a tetracycline antibiotic, has been reported to exert beneficial effects in models of Alzheimer's disease (AD). To characterize the mechanisms underlying the putative minocycline-related neuroprotection, we studied its effect in an in vitro model of AD. Primary hippocampal cultures were treated with β-amyloid peptide (Aβ) and cell viability was assessed by standard MTT-assay. Incubation with 10 μM Aβ for 24 h significantly inhibits cellular MTT-reduction without inducing morphological signs of enhanced cell death or increase in release of lactate dehydrogenase. This indicates that cell viability was not affected. The inhibition of MTT-reduction by Aβ was due to an acceleration of MTT-formazan exocytosis. Intriguingly, the Aβ-triggered increase in MTT-formazan exocytosis was abolished by co-treatment with minocycline. In vehicle-treated cells minocycline had no effect on formazan exocytosis. This hitherto unrecognized property of minocycline has to be noticed in the elucidation of the underlying mechanism of this promising neuroprotectant.

  6. 49 CFR 173.193 - Bromoacetone, methyl bromide, chloropicrin and methyl bromide or methyl chloride mixtures, etc.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... methyl bromide or methyl chloride mixtures, etc. 173.193 Section 173.193 Transportation Other Regulations... bromide, chloropicrin and methyl bromide or methyl chloride mixtures, etc. (a) Bromoacetone must be...) Bromoacetone, methyl bromide, chloropicrin and methyl bromide mixtures, chloropicrin and methyl...

  7. 49 CFR 173.193 - Bromoacetone, methyl bromide, chloropicrin and methyl bromide or methyl chloride mixtures, etc.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... methyl bromide or methyl chloride mixtures, etc. 173.193 Section 173.193 Transportation Other Regulations... bromide, chloropicrin and methyl bromide or methyl chloride mixtures, etc. (a) Bromoacetone must be... Group I performance level. (b) Bromoacetone, methyl bromide, chloropicrin and methyl bromide...

  8. 49 CFR 173.193 - Bromoacetone, methyl bromide, chloropicrin and methyl bromide or methyl chloride mixtures, etc.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... methyl bromide or methyl chloride mixtures, etc. 173.193 Section 173.193 Transportation Other Regulations... bromide, chloropicrin and methyl bromide or methyl chloride mixtures, etc. (a) Bromoacetone must be...) Bromoacetone, methyl bromide, chloropicrin and methyl bromide mixtures, chloropicrin and methyl...

  9. 49 CFR 173.193 - Bromoacetone, methyl bromide, chloropicrin and methyl bromide or methyl chloride mixtures, etc.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... methyl bromide or methyl chloride mixtures, etc. 173.193 Section 173.193 Transportation Other Regulations... bromide, chloropicrin and methyl bromide or methyl chloride mixtures, etc. (a) Bromoacetone must be... Group I performance level. (b) Bromoacetone, methyl bromide, chloropicrin and methyl bromide...

  10. Using Yeast to Easily Determine Mitochondrial Functionality with 1-(4,5-Dimethylthiazol-2-yl)-3,5-Diphenyltetrazolium Bromide (MTT) Assay

    ERIC Educational Resources Information Center

    Sanchez, Norma Silvia; Konigsberg, Mina

    2006-01-01

    Mitochondrial respiration is an important topic in biochemical courses, but at the same time it is a very arduous and challenging subject for the students to comprehend. In teaching laboratories, experiments are also difficult to perform because mitochondrial isolation as well as oxygen consumption determinations require expensive equipment,…

  11. The further application of MTT-formazan colorimetry to studies on filarial worm viability.

    PubMed

    Comley, J C; Townson, S; Rees, M J; Dobinson, A

    1989-09-01

    Experiments have confirmed that MTT-formazan colorimetry in its simplest form (incubation of intact worms with MTT and direct visualisation of any formazan formed) can be readily applied to several species of filariae including Onchocerca volvulus. Data is presented which will assist the development of quantitative MTT reduction viability tests for a selection of the smaller filarial species. Assays of pieces of Onchocerca gutturosa and O. volvulus females have led us to tentatively conclude that the tips of filariae, particularly the anterior ends, may well be metabolically the most active part of the worm. Selective sampling of these regions for Onchocerca might therefore be a useful indicator for the viability of the parasite. An example of how MTT-formazan colorimetry has been applied to yield additional data to support motility observations on the in vitro survival of male O. gutturosa is also given. The in vitro timecourse of worm death caused by 10 microM CGP 20376 on Acanthocheilonema viteae females has been examined by MTT reduction and compared with 6 other non-subjective parameters. The results suggests that the parameters examined could be divided into two groups according to the time taken for CGP 20376 to cause 50% inhibition (t50) of the parameter. Fast response parameters had t50's between 1 and 6 h (motility indices, 14CO2 evolution, adenine uptake and leucine uptake), they are more sensitive measures of viability and indicate possible worm damage which may or may not be reversible. Slow response parameters had t50's between 34 and 48.5 h (lactate output, MTT reduction and adenine leakage), and are probably linked with severe degenerative changes and are indicative of worm death.(ABSTRACT TRUNCATED AT 250 WORDS)

  12. In vitro evaluation of inorganic and methyl mercury mediated cytotoxic effect on neural cells derived from different animal species.

    PubMed

    Tong, Jing; Wang, Youwei; Lu, Yuanan

    2016-03-01

    To extend the current understanding of the mercury-mediated cytotoxic effect, five neural cell lines established from different animal species were comparatively analyzed using three different endpoint bioassays: thiazolyl blue tetrazolium bromide, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay (MTT), neutral red uptake assay (NRU), and Coomassie blue assay (CB). Following a 24-hr exposure to selected concentrations of mercury chloride (HgCl2) and methylmercury (II) chloride (MeHgCl), the cytotoxic effect on test cells was characterized by comparing their 50% inhibition concentration (IC50) values. Experimental results indicated that both these forms of mercury were toxic to all the neural cells, but at very different degrees. The IC50 values of MeHgCl among these cell lines ranged from 1.15±0.22 to 10.31±0.70μmol/L while the IC50 values for HgCl2 were much higher, ranging from 6.44±0.36 to 160.97±19.63μmol/L, indicating the more toxic nature of MeHgCl. The IC50 ratio between HgCl2 and MeHgCl ranged from 1.75 to 96.0, which confirms that organic mercury is much more toxic to these neural cells than inorganic mercury. Among these cell lines, HGST-BR and TriG44 derived from marine sea turtles showed a significantly high tolerance to HgCl2 as compared to the three mammalian neural cells. Among these neural cells, SK-N-SH represented the most sensitive cells to both chemical forms of mercury.

  13. Cytotoxicity and antiangiogenic effects of Rhus coriaria, Pistacia vera and Pistacia khinjuk oleoresin methanol extracts

    PubMed Central

    Mirian, M.; Behrooeian, M.; Ghanadian, M.; Dana, N.; Sadeghi-Aliabadi, H.

    2015-01-01

    Angiogenesis, formation of new blood vessels, play an important role in some diseases such as cancer and its metastasis. Using angiogenesis inhibitors, therefore, is one of the ways for cancer treatment and prevention of metastasis. Medicinal plants have been shown to play a major role in the treatment of a variety of cancers. In this direction, cytotoxic and angiogenic effects of oleo gum resin extracts of Rhus coriaria, Pistacia vera and Pistacia khinjuk from Anacardiaceae family were studied. For IC50 values, cytotoxic effects of the plant extracts were evaluated at different concentrations (1, 10, 20, 40, 80,100 μg/ml) against human umbilical vein endothelial normal cell (HUVEC) and Y79 cell lines using 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. In vitro tube formation on matrigel base was used to evaluate angiogenic effects in the presence of increasing concentrations (50, 100, 250 μg/ml) of the extracts. Vascular endothelium growth factor was used as angiogenesis stimulator. Gas chromatography results showed that α-pinene and β-pinene were the major essential oils constituents of all plant extracts. According to the MTT assay results, the R. coriaria resin extract was more cytotoxic than those of P. vera and P. khinjuk extracts (IC50, 9.1 ± 1.6 vs 9.8 ± 2.1 and 12.0 ± 1.9, respectively; P<0.05). Cytotoxic effects of all extracts against Y79 cell line was significantly higher than those of HUVEC used as a normal cell line (P<0.05). Tube formation assay also showed that extract of R. coriaria resin inhibited angiogenesis more significantly than other tested extracts (P<0.05). It could be concluded that R. coriaria resin extract possess cytotoxic effect and antiangiogenesis against cancer cells and as an anticancer natural product has a good potential for future studies. PMID:26600850

  14. Nymphaea nouchali Burm. f. hydroalcoholic seed extract increases glucose consumption in 3T3-L1 adipocytes through activation of peroxisome proliferator-activated receptor gamma and insulin sensitization

    PubMed Central

    Parimala, Mabel; Debjani, M.; Vasanthi, Hannah Rachel; Shoba, Francis Gricilda

    2015-01-01

    Nymphaea nouchali Burm. f. (Family – Nymphaeaceae) is a well-known medicinal plant used in the Indian ayurvedic system of medicine for treating diabetes. The seeds especially have been prescribed for diabetes. The hydroalcoholic extract of N. nouchali seeds has been demonstrated to possess anti-hyperglycemic effects in diabetic rats, but the functional mechanism remains unknown. The nuclear receptor, peroxisome proliferator-activated receptor gamma (PPARγ) is noted to play an important role in glucose and lipid homeostasis. This study was hence focused in evaluating the effect of the extract on PPARγ activation, adipocyte differentiation, and glucose consumption in 3T3-L1 cells. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), followed by adipogenesis assay using Oil Red O technique. Glucose consumption of preadipocytes and adipocytes in the presence of the extract was also determined. Real-time polymerase chain reaction was performed to identify the expression of genes involved in glucose consumption in the adipocytes. MTT assay confirmed the extract to be nontoxic, and Oil Red O staining confirmed enhanced adipocyte differentiation of 3T3-L1 cells in a dose-dependent manner. The extract also increased the expression of PPARγ target gene, which in turn enhanced the expression of GLUT-4. The data, therefore, suggests that N. nouchali seed extract promotes adipocyte differentiation and glucose consumption by inducing PPARγ activation, which in turn increases mRNA GLUT-4 expression and subsequently enhances insulin-responsiveness in insulin target tissues. PMID:26605160

  15. Nymphaea nouchali Burm. f. hydroalcoholic seed extract increases glucose consumption in 3T3-L1 adipocytes through activation of peroxisome proliferator-activated receptor gamma and insulin sensitization.

    PubMed

    Parimala, Mabel; Debjani, M; Vasanthi, Hannah Rachel; Shoba, Francis Gricilda

    2015-01-01

    Nymphaea nouchali Burm. f. (Family - Nymphaeaceae) is a well-known medicinal plant used in the Indian ayurvedic system of medicine for treating diabetes. The seeds especially have been prescribed for diabetes. The hydroalcoholic extract of N. nouchali seeds has been demonstrated to possess anti-hyperglycemic effects in diabetic rats, but the functional mechanism remains unknown. The nuclear receptor, peroxisome proliferator-activated receptor gamma (PPARγ) is noted to play an important role in glucose and lipid homeostasis. This study was hence focused in evaluating the effect of the extract on PPARγ activation, adipocyte differentiation, and glucose consumption in 3T3-L1 cells. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), followed by adipogenesis assay using Oil Red O technique. Glucose consumption of preadipocytes and adipocytes in the presence of the extract was also determined. Real-time polymerase chain reaction was performed to identify the expression of genes involved in glucose consumption in the adipocytes. MTT assay confirmed the extract to be nontoxic, and Oil Red O staining confirmed enhanced adipocyte differentiation of 3T3-L1 cells in a dose-dependent manner. The extract also increased the expression of PPARγ target gene, which in turn enhanced the expression of GLUT-4. The data, therefore, suggests that N. nouchali seed extract promotes adipocyte differentiation and glucose consumption by inducing PPARγ activation, which in turn increases mRNA GLUT-4 expression and subsequently enhances insulin-responsiveness in insulin target tissues.

  16. Paroxetine Can Enhance Neurogenesis during Neurogenic Differentiation of Human Adipose-derived Stem Cells

    PubMed Central

    Jahromi, Maliheh; Razavi, Shahnaz; Amirpour, Nushin; Khosravizadeh, Zahra

    2016-01-01

    Background: Some antidepressant drugs can promote neuronal cell proliferation in vitro as well as hippocampal neurogenesis in human and animal models. Furthermore, adipose tissue is an available source of adult stem cells with the ability to differentiate in to multiple lineages. Therefore, human Adipose-Derived Stem Cells (hAD-SCs) may be a suitable source for regenerative medical applications. Since there is no evidence for the effect of Paroxetine as the most commonly prescribed antidepressant drug for neurogenic potential of hADSCs, an attempt was made to determine the effect of Paroxetine on proliferation and neural differentiation of hADSCs. Methods: ADSCs were isolated from human abdominal fat. These cells differentiated to neuron-like cells and were treated with Paroxetine. 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay and immunofluorescence technique were used for assessment of cell proliferation and neurogenic differentiation potential of induced cells, respectively. Results: MTT assay analysis showed that Paroxetine significantly increased the proliferation rate of induced hADSCs (p<0.05), while immunofluorescent staining indicated that Paroxetine treatment during neurogenic differentiation could enhance the mean percentage of Nestin and MAP2 (Microtubule-associated protein-2) positive cells but the mean percentage of GFAP (Glial acidic fibrillary protein) positive cells significantly decreased relative to control group (p<0.05). Conclusion: Our results provide evidence that Paroxetine can promote proliferation and differentiation rate during neurogenic differentiation of hADSCs. Moreover, Paroxetine can reduce gliogenesis of induced hADSCs during neurogenic differentiation. PMID:27920882

  17. In vitro cytotoxicity and phototoxicity study of cosmetics colorants.

    PubMed

    Tomankova, K; Kejlova, K; Binder, S; Daskova, A; Zapletalova, J; Bendova, H; Kolarova, H; Jirova, D

    2011-09-01

    The aim of the work was early identification of preventable risk factors connected with the consumers usage of products of everyday use, such as cosmetics, toys and children products, and other materials intended for contact with human skin. The risk factor is represented by substances with irritation potential and subsequent possible sensitisation, resulting in negative impact on human physical and psychical health with social and societal consequences. The legislation for cosmetics, chemical substances and other products requires for hazard identification the application of alternative toxicological methods in vitro without the use of animals. For this reason we used a battery of alternative assays in vitro, based on cell cultures. Progressive methods of molecular biology, based on fluorimetry and fluorescence, were employed for identification of early morphological and functional changes on cellular level. Four colorants frequently used in cosmetics (P-WS Caramel, Chlorophyllin, Unicert Red K 7054-J and Unicert Red K 7008-J) were tested on cell line NIH3T3 (mouse fibroblast cell) and 3T3 Balb/c with/without UV irradiation (dose 5 J cm(-2)). Fluorescence methods for the study of cell damage using fluorescence probes offer results for the evaluation of cytotoxicity and cell viability of adherent cells. We detected intracellular production of ROS investigated by molecular probe CM-H(2)DCFDA, which is primarily sensitive to the increased production of hydrogen peroxide or its downstream products. Toxic effects on the cellular level were identified by viability tests using Neutral Red uptake and MTT assay, where the live cells reduce yellow soluble 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to insoluble formazan crystals. The reaction was investigated on mitochondrial membrane of living cells and the type of cell death was determined using Apoptosis detection kit. Cytotoxicity tests revealed health risks of using Chlorophyllin and Unicert Red

  18. Tellurium tetrachloride and diphenyl ditelluride cause cytotoxicity in rat hippocampal astrocytes.

    PubMed

    Roy, Shalini; Hardej, Diane

    2011-10-01

    Tellurium tetrachloride (TeCl(4)) and diphenyl ditelluride (DPDT) cytotoxicity, was investigated in rat astrocytes. Concentrations of 0.24-250μM (24h) were tested for viability using MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) and trypan blue exclusion. MTT showed significant decreases at all concentrations tested for both compounds. Significant decreases in viability were seen in 1.95-250μM of DPDT and 0.97-250μM of TeCl(4) with trypan blue exclusion. The LC(50) for both compounds was 62.5μM. Light and scanning microscopy confirm toxicity observed at higher concentrations. Thiobarbituric acid reactive substances (TBARs) assay, TUNEL, cytochrome c and caspase release were carried out. No significant increase in TBARS with either agent was observed (15.625-62.5μM). TUNEL and cytochrome c assays demonstrated apoptosis in TeCl(4) treated cells (31.25-125μM). Non-apoptotic cells were observed in DPDT treated cells. Studies of caspase 3/7 and caspase 9 indicated increased activity in TeCl(4) but not in DPDT treated cells. Optical Emission Spectroscopy of DPDT and TeCl(4) treated cells demonstrated significant accumulation of elemental tellurium in all treatment groups (31.25-125μM). We conclude that DPDT and TeCl(4) are cytotoxic to astrocytes. TeCl(4) treated cells die via the intrinsic apoptotic pathway. Accumulation of tellurium occurs with both compounds, but results in different mechanisms of cell death.

  19. Molybdenum nanoparticles-induced cytotoxicity, oxidative stress, G2/M arrest, and DNA damage in mouse skin fibroblast cells (L929).

    PubMed

    Siddiqui, Maqsood A; Saquib, Quaiser; Ahamed, Maqusood; Farshori, Nida N; Ahmad, Javed; Wahab, Rizwan; Khan, Shams T; Alhadlaq, Hisham A; Musarrat, Javed; Al-Khedhairy, Abdulaziz A; Pant, Aditya B

    2015-01-01

    The present investigation was aimed to study the cytotoxicity, oxidative stress, and genotoxicity induced by molybdenum nanoparticles (Mo-NPs) in mouse skin fibroblast cells (L929). Cells were exposed to different concentrations (1-100 μg/ml) of Mo-NPs (size 40 nm) for 24 and 48 h. After the exposure, different cytotoxicity assays (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide, MTT; neutral red uptake, NRU; and cellular morphology) and oxidative stress markers (lipid peroxidation, LPO; glutathione, GSH; and catalase) were studied. Further, Mo-NPs-induced intracellular reactive oxygen species (ROS) generation, mitochondrial membrane potential (MMP), cell cycle arrest, and DNA damage were also studied. L929 cells treated with Mo-NPs showed a concentration- and time-dependent decrease in cell viability and a loss of the normal cell morphology. The percentage cell viability was recorded as 25%, 42%, and 58% by MTT assay and 24%, 46%, and 56% by NRU assay at 25, 50, and 100 μg/ml of Mo-NPs, respectively after 48 h exposure. Furthermore, the cells showed a significant induction of oxidative stress. This was confirmed by the increase in LPO and ROS generation, as well as the decrease in the GSH and catalase levels. The decrease in MMP also confirms the impaired mitochondrial membrane. The cell cycle analysis and comet assay data revealed that Mo-NPs induced G2/M arrest and DNA damage in a concentration-dependent manner. Our results demonstrated, for the first time, Mo-NPs induced cytotoxicity, oxidative stress and genotoxicity in L929 cells. Thus, data suggest the potential hazardous nature of Mo-NPs.

  20. In vitro studies on protective effect of Glycyrrhiza glabra root extracts against cadmium-induced genetic and oxidative damage in human lymphocytes.

    PubMed

    Dirican, Ebubekir; Turkez, Hasan

    2014-01-01

    Cadmium is a modern environmental contaminant that is toxic and carcinogenic. Glycyrrhiza glabra is a traditional medicinal herb which grows in the various parts of the World. Recent studies demonstrated that G. glabra has antifungal, antimicrobial, antioxidant, and powerful antiinflammatory features. The purpose of this study was to investigate the genetic safety of extracts from G. glabra and its effects on cadmium (as CdCl2) induced genotoxicity. Therefore we evaluated the capability of G. glabra extract to inhibit the rate of micronucleus (MN), sister chromatid exchange (SCE) formations induced by CdCl2. Moreover, to assess the effects of G. glabra on cell viability and oxidative status, we performed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and total antioxidant capacity (TAC) assays. Our results showed that there were significant increases (P < 0.05) in both SCE and MN frequencies of cultures treated with CdCl2 (5 ppm) as compared to controls. However, co-application of G. glabra extract (5, 10 and 20 ppm) and CdCl2 resulted in decreases of MN and SCE rates as compared to the group treated with CdCl2 alone. Again, the results of MTT and TAC assays clearly indicated dose dependent ameliorative effects of G. glabra extracts against CdCl2 toxicity. In conclusion, this study demonstrated for the first time that G. glabra extracts provided increased resistance of DNA against CdCl2 induced genetic and oxidative damage in human lymphocytes. So, the risk on target tissues of CdCl2 could be reduced and ensured early recovery from its toxicity.

  1. From the Cover: Vulnerability of C6 Astrocytoma Cells After Single-Compound and Joint Exposure to Type I and Type II Pyrethroid Insecticides.

    PubMed

    Romero, Delfina M; Berardino, Bruno G; Wolansky, Marcelo J; Kotler, Mónica L

    2017-01-01

    A primary mode-of-action of all pyrethroid insecticides (PYRs) is the disruption of the voltage-gated sodium channel electrophysiology in neurons of target pests and nontarget species. The neurological actions of PYRs on non-neuronal cells of the nervous system remain poorly investigated. In the present work, we used C6 astrocytoma cells to study PYR actions (0.1-50 μM) under the hypothesis that glial cells may be targeted by and vulnerable to PYRs. To this end, we characterized the effects of bifenthrin (BF), tefluthrin (TF), α-cypermethrin (α-CYP), and deltamethrin (DM) on the integrity of nuclear, mitochondrial, and lysosomal compartments. In general, 24- to 48-h exposures produced concentration-related impairment of cell viability. In single-compound, 24-h exposure experiments, effective concentration (EC)15s 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT assay) were computed as follows (in μM): BF, 16.1; TF, 37.3; α-CYP, 7.8; DM, 5.0. We found concentration-related damage in several C6-cell subcellular compartments (mitochondria, nuclei, and lysosomes) at ≥ 10(-1) μM levels. Last, we examined a mixture of all PYRs (ie, Σ individual EC15) using MTT assays and subcellular analyses. Our findings indicate that C6 cells are responsive to nM levels of PYRs, suggesting that astroglial susceptibility may contribute to the low-dose neurological effects caused by these insecticides. This research further suggests that C6 cells may provide relevant information as a screening platform for pesticide mixtures targeting nervous system cells by expected and unexpected toxicogenic pathways potentially contributing to clinical neurotoxicity.

  2. Bioactivity and biocompatibility of hydroxyapatite-based bioceramic coatings on zirconium by plasma electrolytic oxidation.

    PubMed

    Aktuğ, Salim Levent; Durdu, Salih; Yalçın, Emine; Çavuşoğlu, Kültigin; Usta, Metin

    2017-02-01

    In the present work, hydroxyapatite (HAP)-based plasma electrolytic oxide (PEO) coatings were produced on zirconium at different current densities in a solution containing calcium acetate and β-calcium glycerophosphate by a single step. The phase structure, surface morphology, functional groups, thickness and roughness of the coatings were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), eddy current method and surface profilometer, respectively. The phases of cubic-zirconia, calcium zirconate and HAP were detected by XRD. The amount of HAP and calcium zirconate increased with increasing current density. The surface of the coatings was very porous and rough. Moreover, bioactivity and biocompatibility of the coatings were analyzed in vitro immersion simulated body fluid (SBF) and MTT (3-(4,5-dimethyl thiazol-2yl)-2,5-diphenyl tetrazolium bromide) assay, hemolysis assay and bacterial formation. The apatite-forming ability of the coatings was evaluated after immersion in SBF up to 28days. After immersion, the bioactivity of HAP-based coatings on zirconium was greater than the ones of uncoated zirconium and zirconium oxide-based surface. The bioactivity of PEO surface on zirconium was significantly improved under SBF conditions. The bacterial adhesion of the coatings decreased with increasing current density. The bacterial adhesion of the coating produced at 0.370A/cm(2) was minimum compared to uncoated zirconium coated at 0.260 and 0.292A/cm(2). The hemocompatibility of HAP-based surfaces was improved by PEO. The cell attachment and proliferation of the PEO coatings were better than the one of uncoated zirconium according to MTT assay results.

  3. Effects of cyclooxygenase-2 on human esophageal squamous cell carcinoma

    PubMed Central

    Zhang, Li; Wu, Yong-Dong; Li, Peng; Tu, Jun; Niu, Ying-Lin; Xu, Cai-Min; Zhang, Shu-Tian

    2011-01-01

    AIM: To study the relationship between the cyclooxygenase (COX)-2 gene and the proliferation and apoptosis of esophageal squamous carcinoma EC109 cells. METHODS: The techniques of RNA interference (RNAi) and cell transfection, as well as the levels of oncogenicity in nude mice, were used to study the role of COX-2 in the esophageal squamous carcinoma cell (ESCC) line EC109. Following RNAi and transfection, Western blotting analysis was used to determine the expression of the COX-2 protein. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) reduction assay was used to evaluate cell growth, and flow cytometry was used to detect cell apoptosis. RESULTS: Western blotting analysis demonstrated that COX-2 expression was significantly reduced in EC109 cells treated with COX-2-specific short interfering RNA (siRNA) but was increased in EC109 cells transfected with COX-2. Furthermore, COX-2 siRNA treatment inhibited cell proliferation (P < 0.01) and induced apoptosis in EC109 cells, as determined by an MTT assay and by flow cytometry, respectively. In contrast, transfected COX-2 led to increased cell proliferation (P < 0.05) and decreased apoptosis in EC109 cells. In addition, combination treatment of cells with COX-2 siRNA and aspirin had a synergistic effect (P < 0.01). For experiments measuring tumorigenicity, xenograft tumors of a greater volume and weight were found in the COX-2 group compared with other groups (P < 0.05). A large dose of aspirin inhibited tumor growth in nude mice effectively (P < 0.05), and the rate of tumor suppression was 51.8% in the high-dose aspirin group. CONCLUSION: COX-2 plays a very critical role in ESCC carcinogenesis, and COX-2 siRNA combined with aspirin has the potential to be an anticancer therapy for the treatment of ESCC. PMID:22147962

  4. Dysregulation of mRNA profile in cisplatin-resistant gastric cancer cell line SGC7901

    PubMed Central

    Xie, Xiao-Que; Zhao, Qi-Hong; Wang, Hua; Gu, Kang-Sheng

    2017-01-01

    AIM To explore novel therapeutic target of cisplatin resistance in human gastric cancer. METHODS The sensitivity of SGC7901 cells and cisplatin-resistant SGC7901 cells (SGC7901/DDP) for cisplatin were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. High-quality total RNA which isolated from SGC7901/DDP cells and SGC7901 cells were used for mRNA microarray analysis. Results were analyzed bioinformatically to predict their roles in the development of cisplatin resistance and the expression of 13 dysregulated mRNAs we selected were validated by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS SGC7901/DDP cells highly resistant to cisplatin demonstrated by MTT assay. A total of 1308 mRNAs (578 upregulated and 730 downregulated) were differentially expressed (fold change ≥ 2 and P-value < 0.05) in the SGC7901/DDP cells compared with SGC7901 cells. The expression of mRNAs detected by qRT-PCR were consistent with the microarray results. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes pathway and protein-protein interaction analysis demonstrated that the differentially expressed mRNAs were enriched in PI3K-Akt, Notch, MAPK, ErbB, Jak-STAT, NF-kappaB signaling pathways which may be involved in cisplatin resistance. Several genes such as PDE3B, VEGFC, IGFBP3, TLR4, HIPK2 and EGF may associated with drug resistance of gastric cancer cells to cisplatin. CONCLUSION Exploration of those altered mRNAs may provide more promising strategy in diagnosis and therapy for gastric cancer with cisplatin resistance. PMID:28275299

  5. Safety assessment of nanoparamagnetic contrast agents with different coatings for molecular MRI

    NASA Astrophysics Data System (ADS)

    Azizian, Gholamreza; Riyahi-Alam, Nader; Haghgoo, Soheila; Saffari, Mojtaba; Zohdiaghdam, Reza; Gorji, Ensieh

    2013-04-01

    Despite the wide application of gadolinium as a contrast agent for magnetic resonance imaging (MRI), there is a serious lack of information on its toxicity. Gadolinium and gadolinium oxide (Gd-oxide) are used as contrast agents for magnetic resonance imaging (MRI). There are methods for reducing toxicity of these materials, such as core nanoparticles coating or conjugating. Therefore, for toxicity evaluation, we compared the viability of commercial contrast agents in MRI (Gd-DTPA) and three nanoparticles with the same core Gd2O3 and small particulate gadolinium oxide or SPGO (< 40 nm) but different coatings of diethyleneglycol (DEG) as Gd2O3-DEG and methoxy polyethylene glycol-silane (mPEG-silane: 550 and 2000 Dalton) as SPGO-mPEG-silane550 and SPGO-mPEG-silane2000, respectively, in the SK-MEL3 cell line, by light microscopy, MTT assay using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide, and the LDH assay detecting lactate dehydrogenase activity. The viability values were not statistically different between the three nanoparticles and Gd-DTPA. The MTT and LDH assay results showed that Gd2O3-DEG nanoparticles were more toxic than Gd-DTPA and other nanoparticles. Also, SPGO-mPEG-silane2000 was more biocompatible than other nanoparticles. The obtained results did not show any significant increase in cytotoxicity of the nanoparticles and Gd-DTPA, neither dose-dependent nor time-dependent. Therefore, DEG and PEG, due to their considerable properties and irregular sizes (different molecular weights), were selected as the useful surface covering materials of nanomagnetic particles that could reveal noticeable relaxivity and biocompatibility characteristics.

  6. Neurological manifestation of methyl bromide intoxication.

    PubMed

    Suwanlaong, Kanokrat; Phanthumchinda, Kammant

    2008-03-01

    Methyl bromide is a highly toxic gas with poor olfactory warning properties. It is widely used as insecticidal fumigant for dry foodstuffs and can be toxic to central and peripheral nervous systems. Most neurological manifestations of methyl bromide intoxication occur from inhalation. Acute toxicity characterized by headache, dizziness, abdominal pain, nausea, vomiting and visual disturbances. Tremor, convulsion, unconsciousness and permanent brain damage may occur in severe poisoning. Chronic exposure can cause neuropathy, pyramidal and cerebellar dysfunction, as well as neuropsychiatric disturbances. The first case of methyl bromide intoxication in Thailand has been described. The patient was a 24-year-old man who worked in a warehouse of imported vegetables fumigated with methyl bromide. He presented with unstable gait, vertigo and paresthesia of both feet, for two weeks. He had a history of chronic exposure to methyl bromide for three years. His fourteen co-workers also developed the same symptoms but less in severity. Neurological examination revealed ataxic gait, decreased pain and vibratory sense on both feet, impaired cerebellar signs and hyperactive reflex in all extremities. The serum concentration of methyl bromide was 8.18 mg/dl. Electrophysilogical study was normal. Magnetic resonance imaging of the brain (MRI) revealed bilateral symmetrical lesion of abnormal hypersignal intensity on T2 and fluid-attenuation inversion recovery (FLAIR) sequences at bilateral dentate nuclei of cerebellum and periventricular area of the fourth ventricle. This incident stresses the need for improvement of worker education and safety precautions during all stages of methyl bromide fumigation.

  7. Antioxidant and Cytotoxic Effect of Barringtonia racemosa and Hibiscus sabdariffa Fruit Extracts in MCF-7 Human Breast Cancer Cell Line

    PubMed Central

    Amran, Norliyana; Rani, Anis Najwa Abdul; Mahmud, Roziahanim; Yin, Khoo Boon

    2016-01-01

    Background: The fruits of Barringtonia racemosa and Hibiscus sabdariffa have been used in the treatment of abscess, ulcer, cough, asthma, and diarrhea as traditional remedy. Objective: This study aims to evaluate cytotoxic effect of B. racemosa and H. sabdariffa methanol fruit extracts toward human breast cancer cell lines (MCF-7) and its antioxidant activities. Materials and Methods: Total antioxidant activities of extracts were assayed using 2,2′-diphenyl-1-picrylhydrazyl radical (DPPH) and β-carotene bleaching assay. Content of phytochemicals, total flavonoid content (TFC), and total phenolic content (TPC) were determined using aluminum chloride colorimetric method and Folin–Ciocalteu's reagent, respectively. Cytotoxic activity in vitro was investigated through 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Results: B. racemosa extract exhibited high antioxidant activities compared to H. sabdariffa methanol fruit extracts in DPPH radical scavenging assay (inhibitory concentration [IC50] 15.26 ± 1.25 μg/mL) and ί-carotene bleaching assay (I% 98.13 ± 1.83%). B. racemosa also showed higher TPC (14.70 ± 1.05 mg gallic acid equivalents [GAE]/g) and TFC (130 ± 1.18 mg quercetin equivalents [QE]/g) compared to H. sabdariffa (3.80 ± 2.13 mg GAE/g and 40.75 ± 1.15 mg QE/g, respectively). In MTT assay, B. racemosa extract also showed a higher cytotoxic activity (IC50 57.61 ± 2.24 μg/mL) compared to H. sabdariffa. Conclusion: The present study indicated that phenolic and flavonoid compounds known for oxidizing activities indicated an important role among the contents of these plants extract. B. racemosa methanol extract have shown potent cytotoxic activity toward MCF-7. Following these promising results, further fractionation of the plant extract is underway to identify important phytochemical bioactives for the development of potential nutraceutical and pharmaceutical use. SUMMARY The phenolic and flavonoid compounds were

  8. Surface and volume properties of dodecylethyldimethylammonium bromide and benzyldimethyldodecylammonium bromide: II. Volumetric properties of dodecylethyldimethylammonium bromide and benzyldimethyldodecylammonium bromide.

    PubMed

    Harkot, Joanna; Jańczuk, Bronisław

    2009-02-15

    Density measurements were carried out for aqueous solutions of two cationic surfactants: dodecylethyldimethylammonium bromide (C(12)(EDMAB)) and benzyldimethyldodecylammonium bromide (BDDAB). On the basis of the obtained results of the measurements the CMC and partial molar volumes of the surfactants studied were also determined. The obtained CMC values were also analyzed with those accounted on the basis of the surface tension data from the previous paper [J. Harkot, B. Jańczuk, J. Colloid Interface Sci. (2008), submitted for publication]. The values of CMC determined from the surface tension and density measurements for C(12)(EDMAB) are equal to 9.9x10(-3) and 1.5x10(-2) M and for BDDAB to 5.25x10(-3) and 5.3x10(-3) M, respectively. These obtained values are very similar. However, in the literature it is difficult to find the CMC values for C(12)(EDMAB) and BDDAB determined by these two methods used by us-especially from the density measurements for BDDAB and surface tension measurements for C(12)(EDMAB). In the case of the apparent molar volumes of C(12)(EDMAB) there is a good agreement between the values obtained by us and those found in the literature. The CMC values for C(12)(EDMAB) and BDDAB were also determined on the basis of their surface tension and free energy of electrostatic interactions between the polar heads of these surfactants and compared with those obtained from the surface tension and density measurements. It was found that the theoretically obtained CMC values were close to those determined from the density and surface tension data for the C(12)(EDMAB) and that the ratios of the CMC values of the surfactants to their concentration at which the water surface tension decreased by about 20 mN/m proved that the presence of the aryl group in the BDDAB head instead of the methyl group caused that its micellization process was more inhibited in relation to its adsorption at air-water interface than that of C(12)(EDMAB).

  9. Particle-induced artifacts in the MTT and LDH viability assays.

    PubMed

    Holder, Amara L; Goth-Goldstein, Regine; Lucas, Donald; Koshland, Catherine P

    2012-09-17

    In vitro testing is a common first step in assessing combustion-generated and engineered nanoparticle-related health hazards. Commercially available viability assays are frequently used to compare the toxicity of different particle types and to generate dose-response data. Nanoparticles, well-known for having large surface areas and chemically active surfaces, may interfere with viability assays, producing a false assessment of toxicity and making it difficult to compare toxicity data. The objective of this study is to measure the extent of particle interference in two common viability assays, the MTT reduction and the lactate dehydrogenase (LDH) release assays. Diesel particles, activated carbon, flame soot, oxidized flame soot, and titanium dioxide particles are assessed for interactions with the MTT and LDH assay under cell-free conditions. Diesel particles, at concentrations as low as 0.05 μg/mL, reduce MTT. Other particle types reduce MTT only at a concentration of 50 μg/mL and higher. The activated carbon, soot, and oxidized soot particles bind LDH to varying extents, reducing the concentration measured in the LDH assay. The interfering effects of the particles explain in part the different toxicities measured in human bronchial epithelial cells (16HBE14o). We conclude that valid particle toxicity assessments can only be assured after first performing controls to verify that the particles under investigation do not interfere with a specific assay at the expected concentrations.

  10. Particle-induced artifacts in the MTT and LDH viability assays

    PubMed Central

    Holder, Amara L.; Goth-Goldstein, Regine; Lucas, Donald; Koshland, Catherine P.

    2012-01-01

    In vitro testing is a common first step in assessing combustion generated and engineered nanoparticle related health hazards. Commercially available viability assays are frequently used to compare the toxicity of different particle types and to generate dose response data. Nanoparticles, well known for having large surface areas and chemically active surfaces, may interfere with viability assays, producing a false assessment of toxicity and making it difficult to compare toxicity data. The objective of this study is to measure the extent of particle interference in two common viability assays, the MTT reduction and the lactate dehydrogenase (LDH) release assays. Diesel particles, activated carbon, flame soot, oxidized flame soot, and titanium dioxide particles are assessed for interactions with the MTT and LDH assay under cell-free conditions. Diesel particles, at concentrations as low as 0.05 μg/ml, reduce MTT. Other particle types reduce MTT only at a concentration of 50 μg/ml and higher. The activated carbon, soot, and oxidized soot particles bind LDH to varying extents, reducing the concentration measured in the LDH assay. The interfering effects of the particles explain in part the different toxicities measured in human bronchial epithelial cells (16HBE14o). We conclude that valid particle toxicity assessments can only be assured after first performing controls to verify that the particles under investigation do not interfere with a specific assay at the expected concentrations. PMID:22799765

  11. Comparison of the MTT1- and MAL31-like maltose transporter genes in lager yeast strains.

    PubMed

    Dietvorst, Judith; Walsh, Mike C; van Heusden, G Paul H; Steensma, H Yde

    2010-09-01

    Maltose transporter genes were isolated from four lager yeast strains and sequenced. All four strains contain at least two different types of maltose transporter genes, MTT1 and MAL31. In addition, 'long' 2.7 kb, and 'short' 2.4 kb, versions of each type exist. The size difference is caused by the insertion of two repeats of 147 bp into the promoter regions of the long versions of the genes. As a consequence of the insertion, two Mal63-binding sites move 294 bp away from the transcription initiation site. The 2.4- and 2.7-kb versions are further highly similar. Only the 2.4-kb versions and not the 2.7-kb versions of MTT1 could restore the rapid growth of lager yeast strain A15 on maltotriose in the presence of antimycin A. These results suggest that insertion of the two repeats into the promoter region of the 'long versions' of MTT1 genes led to a diminished expression of these genes. None of the tested long and short versions of the MAL31 genes were able to restore this growth. As the promoter regions of the MTT1 and MAL31 genes are identical, small differences in the protein sequence may be responsible for the different properties of these genes.

  12. Growth and characterization of lead bromide crystals

    NASA Technical Reports Server (NTRS)

    Singh, N. B.; Gottlieb, M.; Henningsen, T.; Hopkins, R. H.; Mazelsky, R.; Glicksman, M. E.; Coriell, S. R.; Santoro, G. J.; Duval, W. M. B.

    1992-01-01

    Lead(II) bromide was purified by a combination of directional freezing and zone-refining methods. Differential thermal analysis of the lead bromide showed that a destructive phase transformation occurs below the melting temperature. This transformation causes extensive cracking, making it very difficult to grow a large single crystal. Energy of phase transformation for pure lead bromide was determined to be 24.67 cal/g. To circumvent this limitation, crystals were doped by silver bromide which decreased the energy of phase transformation. The addition of silver helped in achieving the size, but enhanced the inhomogeneity in the crystal. The acoustic attenuation constant was almost identical for the pure and doped (below 3000 ppm) crystals.

  13. Emission of methyl bromide from biomass burning

    SciTech Connect

    Manoe, S.; Andreae, M.O. )

    1994-03-04

    Bromine is, per atom, far more efficient than chlorine in destroying stratospheric ozone, and methyl bromide is the single largest source of stratospheric bromine. The two main previously known sources of this compound are emissions from the ocean and from the compound's use as an agricultural pesticide. Laboratory biomass combustion experiments showed that methyl bromide was emitted in the smoke from various fuels tested. Methyl bromide was also found in smoke plumes from wildfires in savannas, chaparral, and boreal forest. Global emissions of methyl bromide from biomass burning are estimated to be in the range of 10 to 50 gigagrams per year, which is comparable to the amount produced by ocean emission and pesticide use and represents a major contribution ([approximately]30 percent) to the stratospheric bromine budget.

  14. A porphyrin-based metal-organic framework as a pH-responsive drug carrier

    NASA Astrophysics Data System (ADS)

    Lin, Wenxin; Hu, Quan; Jiang, Ke; Yang, Yanyu; Yang, Yu; Cui, Yuanjing; Qian, Guodong

    2016-05-01

    A low cytotoxic porphyrin-based metal-organic framework (MOF) PCN-221, which exhibited high PC12 cell viability via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium (MTT) assay, was selected as an oral drug carrier. Methotrexate (MTX) was chosen as the model drug molecule which was absorbed into inner pores and channels of MOFs by diffusion. PCN-221 showed high drug loading and sustained release behavior under physiological environment without "burst effect". The controlled pH-responsive release of drugs by PCN-221 revealed its promising application in oral drug delivery.

  15. Differential hypoxic cytotoxicity of bioreductive agents determined in vitro by the MTT assay

    SciTech Connect

    Stratford, I.J.; Stephens, M.A.

    1989-04-01

    We obtained good agreement between the MTT assay and conventional clonogenic assays regarding the concentration and contact time required to produce a given level of killing of Chinese hamster V79 cells treated in either air of N/sub 2/ with a range of bioreductive cytotoxic drugs. All agents chosen for these experiments represented classes of compounds known to be more toxic towards hypoxic cells than they are to aerobic cells. Namely a quinone, mitomycin C; a di-N-oxide, SR4233; and a number of nitro-heterocyclics including misonidazole. The MTT assay is carried out with V79 cells attached to the bottom of 1 cm glass wells within a 24 well plate. All procedures, that is drug exposure, cell growth, metabolism of MTT are then carried out in situ. To measure optical density we used an ELIZA plate reader modified to take 24-well plates. We propose that this method provides a simple, rapid procedure for evaluating the cytotoxicity of bioreductive drugs.

  16. Cytotoxicity evaluation of Clinacanthus nutans through dimethylthiazol diphenyltetrazolium bromide and neutral red uptake assays

    PubMed Central

    Vajrabhaya, La-Ongthong; Korsuwannawong, Suwanna

    2016-01-01

    Objectives: The aim of this study was to compare the results of dimethylthiazol diphenyltetrazolium bromide (MTT) and neutral red uptake (NRU) assays of Clinacanthus nutans cytotoxicity. Materials and Methods: Mouse fibroblast (L929) cells were exposed to 0.01%, 0.1%, 0.25%, and 0.5% (W/V) C. nutans in a 96-cluster-well-culture plate for 24 h. The cell viability after exposure to C. nutans was determined by MTT and NRU assays in separate tissue culture plates. The two assays were compared through an intra-class correlation coefficient (ICC) analysis. Results: No significant differences in cytotoxicity were noted between the two assays (P > 0.05). The ICC values for agreement between two assays for the negative and positive control groups and C. nutans concentrations of 0.01%, 0.1%, 0.25%, and 0.5% were 0.84, 0.83, 0.77, 0.68, 0.74, and 0.71, respectively. Conclusion: In general, the MTT and NRU assays performed similarly, exhibiting moderate to good correlation for the evaluation of the cytotoxicity of C. nutans. PMID:27011752

  17. Improving the expression of recombinant proteins in E. coli BL21 (DE3) under acetate stress: an alkaline pH shift approach.

    PubMed

    Wang, Hengwei; Wang, Fengqing; Wang, Wei; Yao, Xueling; Wei, Dongzhi; Cheng, Hairong; Deng, Zixin

    2014-01-01

    Excess acetate has long been an issue for the production of recombinant proteins in E. coli cells. Recently, improvements in acetate tolerance have been achieved through the use of genetic strategies and medium supplementation with certain amino acids and pyrimidines. The aim of our study was to evaluate an alternative to improve the acetate tolerance of E. coli BL21 (DE3), a popular strain used to express recombinant proteins. In this work we reported the cultivation of BL21 (DE3) in complex media containing acetate at high concentrations. In the presence of 300 mM acetate, compared with pH 6.5, pH 7.5 improved cell growth by approximately 71%, reduced intracellular acetate by approximately 50%, and restored the expression of glutathione S-transferase (GST), green fluorescent protein (GFP) and cytochrome P450 monooxygenase (CYP). Further experiments showed that alkaline pHs up to 8.5 had little inhibition in the expression of GST, GFP and CYP. In addition, the detrimental effect of acetate on the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) by the cell membrane, an index of cellular metabolic capacity, was substantially alleviated by a shift to alkaline pH values of 7.5-8.0. Thus, we suggest an approach of cultivating E. coli BL21 (DE3) at pH 8.0 ± 0.5 to minimize the effects caused by acetate stress. The proposed strategy of an alkaline pH shift is a simple approach to solving similar bioprocessing problems in the production of biofuels and biochemicals from sugars.

  18. Inhibitory effect of fulvic acid extracted from Canadian sphagnum peat on chemical mediator release by RBL-2H3 and KU812 cells.

    PubMed

    Yamada, Parida; Isoda, Hiroko; Han, Jun Kyu; Talorete, Terence P N; Abe, Yukuo

    2007-05-01

    Fulvic acid (FA) was extracted and purified from Canadian Sphagnum peat (CP-FA) and characterized by using an element analysis meter, Fourier transform infrared (FT-IR) spectroscopy, electron spin resonance (ESR) spectroscopy, and (13)C-nuclear magnetic resonance ((13)C-NMR) spectroscopy. To investigate the antiallergic effect of CP-FA, we incubated rat basophilic leukemia (RBL-2H3) cells with 0.001-10.0 microg/ml of CP-FA and determined the beta-hexosaminidase release inhibition at different response stages. The intracellular calcium [Ca(2+)](i) level was also determined by using Fluo 3-AM, a calcium-specific fluorescent probe, and the cytotoxicity of CP-FA was determined by the 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. The results revealed that RBL-2H3 cells incubated for 48 h with 0.001-10.0 microg/ml of CP-FA did not show any decreased viability. CP-FA inhibited the beta-hexosaminidase release by IgE-sensitized, antigen-stimulated RBL-2H3 cells at the antigen-antibody binding stage and the antibody-receptor binding stage. CP-FA also inhibited histamine release from A23187 plus PMA- or compound 48/80-stimulated KU812 cells. Furthermore, there was a decrease in the intracellular [Ca(2+)](i) level in IgE-sensitized cells incubated with CP-FA and stimulated with antigen. Our results show that CP-FA may be useful for the treatment or prevention of allergic diseases.

  19. NCAM-140 Translocation into Lipid Rafts Mediates the Neuroprotective Effects of GDNF.

    PubMed

    Li, Li; Chen, Huizhen; Wang, Meng; Chen, Fangfang; Gao, Jin; Sun, Shen; Li, Yunqing; Gao, Dianshuai

    2016-03-22

    Glial cell line-derived neurotrophic factor (GDNF) is a potent neurotrophic factor for substantia nigra dopaminergic (DA) neuronal cells. Recent studies have demonstrated that neural cell adhesion molecule functions as a signal transduction receptor for GDNF. The purpose of this study is to reveal whether neural cell adhesion molecule (NCAM) mediates the protective effects of GDNF on DA neuronal cells and further explore the mechanisms involved. We utilized SH-SY5Y cell line to establish a model of 6-hydroxydopamine (6-OHDA)-injured DA neuronal cells. Lentiviral vectors were constructed to knockdown or overexpress NCAM-140, and a density gradient centrifugation method was employed to separate membrane lipid rafts. 3-(4,5-Dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), flow cytometric analysis, and western blotting were used to evaluate the protective effects of GDNF. The results showed that GDNF could protect 6-OHDA-injured SH-SY5Y cells via improving cell viability and decreasing the cell death rate and cleaved caspase-3 expression. NCAM-140 knockdown decreased cell viability and increased the cell death rate and cleaved caspase-3 expression, while its overexpression had the opposite effects. Notably, the amount of NCAM-140 located in lipid rafts increased after GDNF treatment. Pretreatment with 2-bromopalmitate, a specific inhibitor of protein palmitoylation, suppressed NCAM-140 translocation to lipid rafts and reduced the NCAM-mediated protective effects of GDNF on injured DA neuronal cells. Our results suggest that GDNF have the protective effects on injured DA cells by influencing NCAM-140 translocation into lipid rafts.

  20. Cell viability score (CVS) as a good indicator of critical concentration of benzalkonium chloride for toxicity in cultured ocular surface cell lines.

    PubMed

    Iwasawa, Atsuo; Ayaki, Masahiko; Niwano, Yoshimi

    2013-07-01

    Cytotoxicity of benzalkonium chloride (BAK) is a major factor affecting drug cytotoxicity. This study aimed to determine the critical concentration of BAK for cultured ocular cells, using SIRC (rabbit corneal epithelium), BCE C/D-1b (bovine corneal epithelial cells), RC-1 (rabbit corneal epithelium), and Chang (human conjunctival cells). Cell viability was determined following the exposure of cells to 11 concentrations of BAK for 10, 30, or 60 min using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and neutral red assays, and the cell viability score (CVS) was used to evaluate comprehensively the toxicity of BAK. The CVS system consists of two values. The CVS50 was determined by the number of measurements for viability ≥50% of control. The CVS40/80 was calculated as follows: CVS40/80=(number of measurements for viability values >80%)-(number of measurements for viability values <40%). Both %CVS50 and %CVS40/80 decreased with concentrations of BAK. When BAK concentrations were 0.01% or higher, %CVS50 and %CVS40/80 became 0 and less than -90, respectively. Meanwhile, when BAK concentrations were 0.001% or lower, %CVS50 became 100. In the case of %CVS40/80, when the BAK concentrations were 0.002% or lower, the values reached 75 or more, and when 0.0005% or lower, the %CVS40/80 value reached 100. Accordingly, BAK induced very low cytotoxicity in the cultured ocular cell lines at concentrations of 0.002% or lower. The concentration-dependency confirmed that the CVS score is useful for expressing drug cytotoxicity in a simple and comprehensive manner.

  1. Biological evaluation of bismuth non-steroidal anti-inflammatory drugs (BiNSAIDs): stability, toxicity and uptake in HCT-8 colon cancer cells.

    PubMed

    Hawksworth, Emma L; Andrews, Philip C; Lie, Wilford; Lai, Barry; Dillon, Carolyn T

    2014-06-01

    Recent studies showed that the metal-coordinated non-steroidal anti-inflammatory drug (NSAID), copper indomethacin, reduced aberrant crypt formation in the rodent colon cancer model, while also exhibiting gastrointestinal sparing properties. In the present study, the stability and biological activity of three BiNSAIDs of the general formula [Bi(L)3]n, where L=diflunisal (difl), mefenamate (mef) or tolfenamate (tolf) were examined. NMR spectroscopy of high concentrations of BiNSAIDs (24h in cell medium, 37°C) indicated that their structural stability and interactions with cell medium components were NSAID specific. Assessment of cell viability using the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium]bromide (MTT) assay showed that the toxicity ranking of the BiNSAIDs paralleled those of the respective free NSAIDs: diflH

  2. In vitro cytotoxicity of Selol-loaded magnetic nanocapsules against neoplastic cell lines under AC magnetic field activation

    NASA Astrophysics Data System (ADS)

    Falqueiro, A. M.; Siqueira-Moura, M. P.; Jardim, D. R.; Primo, F. L.; Morais, P. C.; Mosiniewicz-Szablewska, E.; Suchocki, P.; Tedesco, A. C.

    2012-04-01

    The goals of this study are to evaluate invitro compatibility of magnetic nanomaterials and their therapeutic potential against cancer cells. Highly stable ionic magnetic fluid sample (maghemite, γ-Fe2O3) and Selol were incorporated into polymeric nanocapsules by nanoprecipitation method. The cytotoxic effect of Selol-loaded magnetic nanocapsules was assessed on murine melanoma (B16-F10) and oral squamous cell carcinoma (OSCC) cell lines following AC magnetic field application. The influence of different nanocapsules on cell viability was investigated by colorimetric MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. In the absence of AC magnetic field Selol-loaded magnetic nanocapsules, containing 100 µg/mL Selol plus 5 × 1012 particle/mL, showed antitumoral activity of about 50% on B16-F10 melanoma cells while OSCC carcinoma cells demonstrated drug resistance at all concentrations of Selol and magnetic fluid (range of 100-500 µg/mL Selol and 5 × 1012-2.5 × 1013 particle/mL). On the other hand, under AC applied fields (1 MHz and 40 Oe amplitude) B16-F10 cell viability was reduced down to 40.5% (±3.33) at the highest concentration of nanoencapsulated Selol. The major effect, however, was observed on OSCC cells since the cell viability drops down to about 33.3% (±0.38) under application of AC magnetic field. These findings clearly indicate that the Selol-loaded magnetic nanocapsules present different toxic effects on neoplastic cell lines. Further, the cytotoxic effect was maximized under AC magnetic field application on OSCC, which emphasizes the effectiveness of the magnetohyperthermia approach.

  3. Growth Inhibition and Apoptosis Induction by (+)-Cyanidan-3-ol in Hepatocellular Carcinoma

    PubMed Central

    Monga, Jitender; Pandit, Saurabh; Chauhan, Rajinder Singh; Chauhan, Chetan Singh; Chauhan, Shailender Singh; Sharma, Manu

    2013-01-01

    The objective of this study was to evaluate the cytotoxicity of (+)-cyanidan-3-ol (CD-3) in human hepatocellular carcinoma cell line (HepG2) and chemopreventive potential against hepatocellular carcinoma (HCC) in Balb/c mice. The HepG2 cell line was treated with CD-3 at various concentrations and the proliferation of the HepG2 cells was measure by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT), sulforhodamine B (SRB) and lactate dehydrogenase (LDH) assays. Cell apoptosis was detected by Hoechst 33258 (HO), Acridine orange/ethylene dibromide (AO/EB) staining, DNA fragmentation analysis and the apoptosis rate was detected by flow cytometry. The HCC tumor model was established in mice by injecting N-nitrosodiethylamine/carbon tetrachloride (NDEA/CCl4) and the effect of CD-3 on tumor growth in-vivo was studied. The levels of liver injury markers, tumor markers, and oxidative stress were measured. The expression levels of apoptosis-related genes in in-vitro and in vivo models were determined by RT-PCR and ELISA. The CD-3 induced cell death was considered to be apoptotic by observing the typical apoptotic morphological changes under fluorescent microscopy and DNA fragmentation analysis. Annexin V/PI assay demonstrated that apoptosis increased with increase in the concentration of CD-3. The expression levels of apoptosis-related genes that belong to bcl-2 and caspase family were increased and AP-1 and NF-κB activities were significantly suppressed by CD-3. Immunohistochemistry data revealed less localization of p53, p65 and c-jun in CD-3 treated tumors as compared to localization in NDEA/CCl4 treated tumors. Taken together, our data demonstrated that CD-3 could significantly inhibit the proliferation of HepG2 cells in-vitro and suppress HCC tumor growth in-vivo by apoptosis induction. PMID:23894334

  4. Highly efficient synthetic iron-dependent nucleases activate both intrinsic and extrinsic apoptotic death pathways in leukemia cancer cells.

    PubMed

    Horn, Adolfo; Fernandes, Christiane; Parrilha, Gabrieli L; Kanashiro, Milton M; Borges, Franz V; de Melo, Edésio J T; Schenk, Gerhard; Terenzi, Hernán; Pich, Claus T

    2013-11-01

    The nuclease activity and the cytotoxicity toward human leukemia cancer cells of iron complexes, [Fe(HPClNOL)Cl2]NO3 (1), [Cl(HPClNOL)Fe(μ-O)Fe(HPClNOL)Cl]Cl2·2H2O (2), and [(SO4)(HPClNOL)Fe(μ-O)Fe(HPClNOL)(SO4)]·6H2O (3) (HPClNOL=1-(bis-pyridin-2-ylmethyl-amino)-3-chloropropan-2-ol), were investigated. Each complex was able to promote plasmid DNA cleavage and change the supercoiled form of the plasmid to circular and linear ones. Kinetic data revealed that (1), (2) and (3) increase the rate of DNA hydrolysis about 278, 192 and 339 million-fold, respectively. The activity of the complexes was inhibited by distamycin, indicating that they interact with the minor groove of the DNA. The cytotoxic activity of the complexes toward U937, HL-60, Jukart and THP-1 leukemia cancer cells was studied employing 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), fluorescence and electronic transmission microscopies, flow cytometry and a cytochrome C release assay. Compound (2) has the highest activity toward cancer cells and is the least toxic for normal ones (i.e. peripheral blood mononuclear cells (PBMCs)). In contrast, compound (1) is the least active toward cancer cells but displays the highest toxicity toward normal cells. Transmission electronic microscopy indicates that cell death shows features typical of apoptotic cells, which was confirmed using the annexin V-FITC/PI (fluorescein isothiocyanate/propidium iodide) assay. Furthermore, our data demonstrate that at an early stage during the treatment with complex (2) mitochondria lose their transmembrane potential, resulting in cytochrome C release. A quantification of caspases 3, 9 (intrinsic apoptosis pathway) and caspase 8 (extrinsic apoptosis pathway) indicated that both the intrinsic (via mitochondria) and extrinsic (via death receptors) pathways are involved in the apoptotic stimuli.

  5. A Novel Hsp90 Inhibitor Activates Compensatory Heat Shock Protein Responses and Autophagy and Alleviates Mutant A53T α-Synuclein Toxicity.

    PubMed

    Xiong, Rui; Zhou, Wenbo; Siegel, David; Kitson, Russell R A; Freed, Curt R; Moody, Christopher J; Ross, David

    2015-12-01

    A potential cause of neurodegenerative diseases, including Parkinson's disease (PD), is protein misfolding and aggregation that in turn leads to neurotoxicity. Targeting Hsp90 is an attractive strategy to halt neurodegenerative diseases, and benzoquinone ansamycin (BQA) Hsp90 inhibitors such as geldanamycin (GA) and 17-(allylamino)-17-demethoxygeldanamycin have been shown to be beneficial in mutant A53T α-synuclein PD models. However, current BQA inhibitors result in off-target toxicities via redox cycling and/or arylation of nucleophiles at the C19 position. We developed novel 19-substituted BQA (19BQA) as a means to prevent arylation. In this study, our data demonstrated that 19-phenyl-GA, a lead 19BQA in the GA series, was redox stable and exhibited little toxicity relative to its parent quinone GA in human dopaminergic SH-SY5Y cells as examined by oxygen consumption, trypan blue, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), and apoptosis assays. Meanwhile, 19-phenyl-GA retained the ability to induce autophagy and potentially protective heat shock proteins (HSPs) such as Hsp70 and Hsp27. We found that transduction of A53T, but not wild type (WT) α-synuclein, induced toxicity in SH-SY5Y cells. 19-Phenyl-GA decreased oligomer formation and toxicity of A53T α-synuclein in transduced cells. Mechanistic studies indicated that mammalian target of rapamycin (mTOR)/p70 ribosomal S6 kinase signaling was activated by A53T but not WT α-synuclein, and 19-phenyl-GA decreased mTOR activation that may be associated with A53T α-synuclein toxicity. In summary, our results indicate that 19BQAs such as 19-phenyl-GA may provide a means to modulate protein-handling systems including HSPs and autophagy, thereby reducing the aggregation and toxicity of proteins such as mutant A53T α-synuclein.

  6. Binary and tertiary combination of alternariol, 3-acetyl-deoxynivalenol and 15-acetyl-deoxynivalenol on HepG2 cells: Toxic effects and evaluation of degradation products.

    PubMed

    Juan-García, Ana; Juan, Cristina; Manyes, Lara; Ruiz, María-José

    2016-08-01

    Fungi producers of mycotoxins are able to synthesize more than one toxin. Alternariol (AOH) is one of the mycotoxins produced by several Alternaria species, the most common one being Alternaria alternata. The toxins 3-Acetyl-deoxynivalenol (3-ADON) and 15-Acetyl-deoxynivalenol (15-ADON) are acetylated forms of deoxynivalenol (DON) produced by Fusarium graminearum. In the present work it is determined and evaluated the toxic effects of binary and tertiary combination treatment of HepG2 cells with AOH, 3-ADON and 15-ADON, by using the MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide), to subsequently apply the isobologram method and elucidate if the mixtures of these mycotoxins produced synergism, antagonism or additive effect; and lastly, to analyze mycotoxins conversion into metabolites produced and released by HepG2 cells after applying the treatment conditions by liquid chromatography tandem mass spectrometry (LC-MS/MS) equipment and extracted from culture media. HepG2 cells were treated at different concentrations over 24, 48 and 72h. IC50 values detected at all times assayed, ranged from 0.8 to >25μM in binary combinations; while in tertiary it ranged from 7.5 to 12μM. Synergistic, antagonism or additive effect detected in the mixtures of these mycotoxins was different depending on low or high concentration. Among all four mycotoxins combinations assayed, 15-ADON+3-ADON presented the highest toxic potential. At all assayed times, recoveries values oscillated depending on the time and combination studied.

  7. Quaternized chitosan oligomers as novel gene delivery vectors in epithelial cell lines.

    PubMed

    Thanou, M; Florea, B I; Geldof, M; Junginger, H E; Borchard, G

    2002-01-01

    Quaternized modifications of chitosan present characteristics that might be useful in DNA condensing and efficient gene delivery. Trimethylated chitosan (TMO) was synthesized from oligomeric chitosan (<20 monomer units). TMOs spontaneously formed complexes (chitoplexes) with RSV-alpha3 luciferase plasmid DNA. These complexes were characterized by photon correlation spectroscopy and were investigated for their ability to transfect COS-1 and Caco-2 cell lines in the presence and absence of fetal calf serum and compared with DOTAP (N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium sulphate) lipoplexes. Additionally, their effect on the viability of the respective cell cultures was investigated using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. Results showed that quaternized chitosan oligomers were able to condense DNA and form complexes with a size ranging from 200 to 500 nm. Chitoplexes proved to transfect COS-1 cells, however, to a lesser extent than DOTAP-DNA lipoplexes. The quaternized oligomer derivatives appeared to be superior to oligomeric chitosan. The presence of fetal calf serum (FCS) did not affect the transfection efficiency of the chitoplexes, whereas the transfection efficiency of DOTAP DNA complexes was decreased. Cells remained 100% viable in the presence of chitosan oligomers whereas viability of DOTAP treated cells decreased to approximately 50% in both cell lines. Both DOTAP-DNA lipoplexes and chitoplexes resulted in less transfection efficiency in Caco-2 cell cultures than in COS-1 cells; however quaternized chitosan oligomers proved to be superior to DOTAP. Effects on the viability of Caco-2 cells were similar to the effects observed in COS-1 cells. We conclude that trimethylated chitosan-DNA complexes present suitable characteristics and the potential to be used as gene delivery vectors.

  8. Synthesis and molecular docking against dihydrofolate reductase of novel pyridin-N-ethyl-N-methylbenzenesulfonamides as efficient anticancer and antimicrobial agents

    NASA Astrophysics Data System (ADS)

    Debbabi, Khaled F.; Bashandy, Mahmoud S.; Al-Harbi, Sami A.; Aljuhani, Enas H.; Al-Saidi, Hamed M.

    2017-03-01

    This article describes the synthesis of some novel sulfonamides having biologically active pyridine 21-28. Starting with 4-(1-(2-(2-cyanoacetyl)hydrazono)ethyl)-N-ethyl-N-methylbenzenesulfonamide (2), which was prepared from condensation of acetophenone derivative 1 with 2-cyanoacetohydrazide. Interaction of compound 2 with different aldehydes namely 4-fluorobenzaldehyde, 4-hydroxybenzaldehyde and 4-N,N-dimethylbenzaldehyde afforded the corresponding hydrazono-ethyl-N-ethyl-N-methylbenzene sulfonamides 18-20 respectively, which when reacted with malononitrile and ethyl cyanoacetate afforded compounds 21-26 respectively. These compounds 21-26 can be prepared by another reaction route by interaction of compounds 2 with arylidine malononitrile and arylidine ethyl cyanoacetate in refluxing dioxane in the presence of trimethylamine as catalyst. Interaction of compound 2 with malononitrile and ethyl cyanoacetate afforded oxopyridine derivatives 27 and 28 respectively. All the new prepared compounds were evaluated for their antitumor activities against the cell lines MCF-7 in comparison with the reference drug Doxorubicin using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay. Compounds 25, 21, 23 with SI values of 9.72, 9.71, 8.81 respectively, exhibited better activity than doxorubicin (Dox) as a reference drug with SI value of 8.49. In addition, compounds 25, 27 and 22 exhibited anti-bacterial activity against gram-negative bacteria (Klebsiella pneumoniae) with inhibition zones 22.6, 20.3 and 19.3 mm respectively, which were more active than gentamicin as a reference drug with inhibition zone 17.3 mm. Molecular Operating Environment (MOE) performed virtual screening using molecular docking studies of the synthesized compounds. The results indicated that some synthesized compounds suitable inhibitor against dihydrofolate reductase (DHFR) enzyme (PDB SD: 4DFR) with further modification.

  9. Areca nut extract inhibits the growth, attachment, and matrix protein synthesis of cultured human gingival fibroblasts.

    PubMed

    Chang, M C; Kuo, M Y; Hahn, L J; Hsieh, C C; Lin, S K; Jeng, J H

    1998-10-01

    Betel quid chewing is a popular oral habit in India, South Africa, and many Southeast Asian countries. The effects of areca nut (AN) extract on the growth, attachment, and protein synthesis of healthy human gingival fibroblasts (GF) were investigated to determine why betel quid (BQ) chewers have higher prevalence of periodontal disease than non-chewers. Twenty-four hour exposure of human GF to AN extract (> 200 microg/ml) in culture led to the formation of numerous intracellular vacuoles. As analyzed by modified MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] assay, AN extract significantly suppressed the growth of GF over 5 days of incubation in a dose-dependent manner. At concentrations of 50 and 300 microg/ml, AN extract suppressed the growth of GF with 30% and 57% (P < 0.05), respectively. AN extract also significantly suppressed the synthesis of [3H]proline incorporation into trichloroacetic acid (TCA) precipitated proteins. At concentrations of 200, 400, and 600 microg/ml, AN extract suppressed the protein synthesis with 33%, 58%, and 63% of inhibition (P < 0.05), respectively. Preincubation of cells in a medium containing AN extract for 2 hours inhibits the subsequent attachment of cultured GF to type I collagen at the 50% inhibitory concentration (IC50) which is about 720 to 798 microg/ml. Considering the frequent consumption of BQ throughout the day, impairment of sequential fibroblast functions by BQ ingredients is a potential mechanism through which BQ chewing exert a deleterious effect to the gingival tissues.

  10. Effect of surface roughness of ground titanium on initial cell adhesion.

    PubMed

    Huang, Her-Hsiung; Ho, Chun-Te; Lee, Tzu-Hsin; Lee, Tien-Ling; Liao, Ko-Kaung; Chen, Fang-Lung

    2004-11-01

    The effect of surface roughness of ground Ti on the initial adhesion of osteoblast-like U-2 OS cells was investigated in this study. Different numbers (#120, #600, and #1500) of SiC sandpaper and two Al2O3 polishing powder (0.3 and 1 microm) were used to prepare the metal specimens with varying degrees of surface roughness. Surface roughness (Ra) was measured by profilometry. Surface topography was observed using an atomic force microscope. MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay was used to measure the optical density (OD) of specimens after 2 h of cell incubation. The OD value was analyzed by one-way ANOVA for analyzing the factor of surface roughness. Crystal violet staining technique was used to characterize the cell spreading. Results showed that the specimen of #1500 Ti (Ra: 0.15 microm) had the highest OD value. The specimens polished with 0.3 and 1 microm Al2O3 powder (Ra: 0.05 and 0.07 microm) exhibited the worst cell adhesion behavior. Contact guidance of cells could be observed on the rougher #600 and #120 specimens (Ra: 0.33 and 1.20 microm). This study concludes that the surface roughness (Ra: 0.05-1.20 microm) of ground Ti has a highly significant influence on the initial adhesion of osteoblast-like U-2 OS cells. The ground Ti with an Ra of 0.15 microm shows the optimal cell adhesion behavior with respect to either the rougher or smoother specimens.

  11. Cytotoxic effects of Pinus eldarica essential oil and extracts on HeLa and MCF-7 cell lines

    PubMed Central

    Sarvmeili, Najmeh; Jafarian-Dehkordi, Abbas; Zolfaghari, Behzad

    2016-01-01

    Several attempts have so far been made in the search of new anticancer agents of plant origin. Some studies have reported that different species of Pine genus possess cytotoxic activities against various cancer cell lines. In the present study, we evaluated the cytotoxic effects of Pinus eldarica bark and leaf extracts or leaf essential oil on HeLa and MCF-7 tumor cell lines. Hydroalcoholic and phenolic extracts and the essential oil of plant were prepared. Total phenolic contents of the extracts were measured using Folin-Ciocalteu reagent. Essential oil components were determined by gas chromatography-mass spectroscopy (GC-MS). Cytotoxic activity of the extracts and essential oil against HeLa and MCF-7 tumor cell lines were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The polyphenolic content of hydroalcoholic and phenolic extracts of the bark and hydroalcoholic extract of the leaf were 48.31%, 47.2%, and 8.47%, respectively. According to the GC-MS analysis, the major components of the leaf oil of P. eldarica were: β -caryophyllene (14.8%), germacrene D (12.9%), α–terpinenyl acetate (8.15%), α –pinene (5.7%), and –α humulene (5.9%). Bark extracts and leaf essential oil of P. eldarica significantly reduced the viability of both HeLa and MCF-7 cells in a concentration dependent manner. However, leaf extract showed less inhibitory effects against both cell lines. The essential oil of P. eldarica was more cytotoxic than its hydroalcoholic and phenolic extracts. The terpenes and phenolic compounds were probably responsible for cytotoxicity of P. eldarica. Therefore, P. eldarica might have a good potential for active anticancer agents. PMID:28003841

  12. Study the cytotoxicity of different kinds of water-soluble nanoparticles in human osteoblast-like MG-63 cells

    SciTech Connect

    Niu, Lu; Li, Yang; Li, Xiaojie; Gao, Xue; Su, Xingguang

    2012-11-15

    Highlights: ► Preparation of three kinds of water-soluble QDs: CdTe, CdTe@SiO{sub 2}, Mn:ZnSe. ► Evaluated the cytotoxicity qualitatively and quantitatively. ► Fluorescent staining. ► Detected the total intracellular cadmium in cells. -- Abstract: Quantum nanoparticles have been applied extensively in biological and medical fields, the cytotoxicity of nanoparticles becomes the key point we should concern. In this paper, the cytotoxicity of three kinds of water-soluble nanoparticles: CdTe, CdTe@SiO{sub 2} and Mn:ZnSe was studied. We evaluated the nanoparticles toxicity qualitatively by observing the morphological changes of human osteoblast-like MG-63 cells at different incubation times and colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays were carried out to detect the cell viability quantitatively. The results showed that CdTe nanoparticles with high concentrations caused cells to die largely while CdTe@SiO{sub 2} and Mn:ZnSe nanoparticles had no obvious effect. For further study, we studied the relation between the cell viability and the total cadmium concentration in cells and found that the viability of cells treated with CdTe@SiO{sub 2} nanoparticles was higher than that treated with CdTe nanoparticles. We also discovered that the death rate of cells co-incubated with CdTe nanoparticles was proportional to the total intracellular cadmium concentrations.

  13. Lanthanide-Connecting and Lone-Electron-Pair Active Trigonal-Pyramidal-AsO3 Inducing Nanosized Poly(polyoxotungstate) Aggregates and Their Anticancer Activities

    NASA Astrophysics Data System (ADS)

    Zhao, Jun-Wei; Li, Hai-Lou; Ma, Xing; Xie, Zhigang; Chen, Li-Juan; Zhu, Yongsheng

    2016-05-01

    By virtue of the stereochemical effect of the lone-electron pair located on the trigonal-pyramidal-AsO3 groups and the one-pot self-assembly strategy in the conventional aqueous solution, a series of novel lanthanide-bridging and lone-electron-pair active trigonal-pyramidal-AsO3 inducing nanosized poly(polyoxotungstate) aggregates [H2N(CH3)2]6 Na24H16{[Ln10W16(H2O)30O50](B-α-AsW9O33)8}·97H2O [Ln = EuIII (1), SmIII (2), GdIII (3), TbIII (4), DyIII (5), HoIII (6), ErIII (7), TmIII (8)] were prepared and further characterized by elemental analyses, IR spectra, UV spectra, thermogravimetric (TG) analyses and single-crystal X-ray diffraction. The most remarkable structural feature is that the polyanionic skeleton of {[Ln10W16(H2O)30O50](B-α-AsW9O33)8}46‑ is constructed from eight trivacant Keggin [B-α-AsW9O33]9‑ fragments through ten Ln centers and sixteen bridging W atoms in the participation of fifty extraneous oxygen atoms. Notably, 4 and 8 can be stable in the aqueous solution not only for eight days but also in the range of pH = 3.9–7.5. Moreover, the cytotoxicity tests of 4 and 8 toward human cervical cancer (HeLa) cells, human breast cancer (MCF–7) cells and mouse fibroblast (L929) cells were performed by the 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and the cell apoptosis processes were characterized by calcein AM/PI staining experiments, annexin V-FITC/PI staining experiments and morphological changes.

  14. Active targeting behaviors of biotinylated pluronic/poly(lactic acid) nanoparticles in vitro through three-step biotin-avidin interaction.

    PubMed

    Xiong, Xiang Yuan; Gong, Yan Chun; Li, Zi Ling; Li, Yu Ping; Guo, Liang

    2011-01-01

    In order to prepare targeted drug carriers, previously a biotin group has been attached by our group to the end of Pluronic F87/poly(lactic acid) and Pluronic P85/poly(lactic acid) block co-polymers to obtain B-F87-PLA and B-P85-PLA, respectively. In this paper, the active targeting properties of B-F87-PLA and B-P85-PLA nanoparticles in vitro were investigated through a three-step biotin-avidin interaction by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) tests and fluorescence microscopy (FM). Two kinds of human ovarian cancer cells (OVCAR-3 and SKOV-3) and paclitaxel were chosen for the cytotoxicity tests. CA-125 antigen is over-expressed on OVCAR-3 cells but not on SKOV-3 cells. The loading and release behavior of paclitaxel loaded in B-Pluronic-PLA nanoparticles were also studied. Paclitaxel loaded in both B-F87-PLA and B-P85-PLA nanoparticles shows an initial rapid release followed by a slow release period. Compared with SKOV-3 cells, the cytotoxicity results implied that paclitaxel-loaded B-Pluronic-PLA nanoparticles were delivered more effectively to OVCAR-3 cells due to the specific interaction between the biotin groups on the surface of B-Pluronic-PLA nanoparticles and the avidin/biotinylated MAb X306/CA-125 antigen complexes on the surface of OVCAR-3 cells. The active targeting properties of B-F87-PLA nanoparticles were further confirmed by FM.

  15. Chitosan-mediated siRNA delivery in vitro: effect of polymer molecular weight, concentration and salt forms.

    PubMed

    Techaarpornkul, Sunee; Wongkupasert, Sirirat; Opanasopit, Praneet; Apirakaramwong, Auayporn; Nunthanid, Jurairat; Ruktanonchai, Uracha

    2010-03-01

    The aim of this study was to investigate chitosan/siRNA complexes formulated with various chitosan salts (CS) including chitosan aspartate (CS-Asp), chitosan glutamate (CS-Glu), chitosan acetate (CS-Ac), and chitosan hydrochloride (CS-HCl) for in vitro siRNA delivery into stable and constitutive enhanced green fluorescent protein (EGFP)-expressing HeLa cells. The CS/siRNA complexes were characterized by 2% agarose gel electrophoresis and investigated for their transfection efficiency in stable and constitutive EGFP-expressing HeLa cells. The cytotoxicity of the complexes was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The formation of complexes CS/siRNA is mainly dependent on the weight ratio, whereas salt form and molecular weight has less effect. The particle sizes of the complete complexes were in the range of 270-373 nm except the complete complex of CS-Ac, with a slightly positive charge of less than 2 mV. The ability of CS to transfer functionally active siRNA into cell culture is mainly dependent on the weight ratio and molecular weight of CS whereas salt form of CS has less effect. The high gene-silencing efficiency was observed with low MW of CS (20 kDa) and high weight ratio of 32. Over 80% average cell viabilities were observed for CS/siRNA complexes in all weight ratios comparison to untreated cells. This study suggests CS salts have the potential to be used as safe siRNA delivery vectors.

  16. Moxidectin causes adult worm mortality of human lymphatic filarial parasite Brugia malayi in rodent models.

    PubMed

    Verma, Meenakshi; Pathak, Manisha; Shahab, Mohd; Singh, Kavita; Mitra, Kalyan; Misra-Bhattacharya, Shailja

    2014-12-01

    Moxidectin is a macrocyclic lactone belonging to milbemycin family closely related to ivermectin and is currently progressing towards Phase III clinical trial against human infection with the filaria Onchocerca volvulus (Leuckart, 1894). There is a single report on the microfilaricidal and embryostatic activity of moxidectin in case of the human lymphatic filarial parasite Brugia malayi (Brug, 1927) in Mastomys coucha (Smith) but without any adulticidal action. In the present study, the in vitro and in vivo antifilarial efficacy of moxidectin was evaluated on, B. malayi. In vitro moxidectin showed 100% reduction in adult female worm motility at 0.6 μM concentration within 7 days with 68% inhibition in the reduction of MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide dye) (which is used to detect viability of worms). A 50% inhibitory concentration (IC50) of moxidectin for adult female parasite was 0.242 μM, for male worm 0.186 μM and for microfilaria IC50 was 0.813 μM. In adult B. malayi-transplanted primary screening model (Meriones unguiculatus Milne-Edwards), moxidectin at a single optimal dose of 20 mg/kg by oral and subcutaneous route was found effective on both adult parasites and microfilariae. In secondary screening (M coucha, subcutaneously inoculated with infective larvae), moxidectin at the same dose by subcutaneous route brought about death of 49% of adult worms besides causing sterilisation in 54% of the recovered live female worms. The treated animals exhibited a continuous and sustained reduction in peripheral blood microfilaraemia throughout the observation period of 90 days. The mechanism of action of moxidectin is suggested to be similar to avermectins. The in silico studies were also designed to explore the interaction of moxidectin with glutamate-gated chloride channels of B. malayi. The docking results revealed a close interaction of moxidectin with various GluCl ligand sites of B. malayi.

  17. Antitumour and anti-inflammatory effects of palladium(II) complexes on Ehrlich tumour.

    PubMed

    Quilles, Marcela B; Carli, Camila B A; Ananias, Sandra R; Ferreira, Lucas S; Ribeiro, Livia C A; Maia, Danielle C G; Resende, Flávia A; Moro, Antônio C; Varanda, Eliana A; Placeres, Marisa Campos Polesi; Mauro, Antonio E; Carlos, Iracilda Z

    2013-01-01

    Palladium(II) complexes are an important class of cyclopalladated compounds that play a pivotal role in various pharmaceutical applications. Here, we investigated the antitumour, anti-inflammatory, and mutagenic effects of two complexes: [Pd(dmba)(Cl)tu] (1) and [Pd(dmba)(N3)tu] (2) (dmba = N,N-dimethylbenzylamine and tu = thiourea), on Ehrlich ascites tumour (EAT) cells and peritoneal exudate cells (PECs) from mice bearing solid Ehrlich tumour. The cytotoxic effects of the complexes on EAT cells and PECs were assessed using the 3-(4,5-dimethylthiazol-3-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. The effects of the complexes on the immune system were assessed based on the production of nitric oxide (NO) (Griess assay) and tumour necrosis factor-alpha (TNF-alpha), interleukin-12 (IL-12), and interleukin-10 (IL-10) (ELISA). Finally the mutagenic activity was assessed by the Ames test using the Salmonella typhimurium strain TA 98. Cisplatin was used as a standard. The IC50 ranges for the growth inhibition of EAT cells and PECs were found to be (72.8 +/- 3.23) microM and (137.65 +/- 0.22) microM for 1 and (39.7 +/- 0.30) microM and (146.51 +/- 2.67) microM for 2, respectively. The production of NO, IL-12, and TNF-alpha, but not IL-10, was induced by both complexes and cisplatin. The complexes showed no mutagenicity in vitro, unlike cisplatin, which was mutagenic in the strain. These results indicate that the complexes are not mutagenic and have potential immunological and antitumour activities. These properties make them promising alternatives to cisplatin.

  18. In vitro antioxidant and cytotoxic potential of different parts of Tephrosia purpurea

    PubMed Central

    Padmapriya, Ramamoorthy; Ashwini, Sankar; Raveendran, Ramasamy

    2017-01-01

    The antioxidant and cytotoxic properties of four major parts of methanolic extracts of Tephrosia purpurea including leaves, root, stem and seed were investigated and compared. In vitro antioxidant activity of T. purpurea extracts was evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), reducing power assay and antihemolytic assay. In vitro cytotoxic effect of T. purpurea extracts on SW620 colorectal cancer cell line was studied using 3-(4, 5-dimethylthiazolyl -2,5-diphenyl-tetrazolium bromide (MTT) assay. Folin-ciocalteu and aluminium chloride methods were used to determine the total phenolic and flavonoid contents respectively. Among the four extracts studied, leaves extract showed the highest antioxidant activity, DPPH: 186.3 ± 14.0 μg/mL, FRAP: 754.2 ± 50.9 μmol Fe(II)/mg and reducing power activity: 65.7 ± 4.2 μg/mg of quercetin equivalent (QE/mg) and there was no significant difference observed in antihemolytic activity. Leaves extract showed effective cytotoxicity on colorectal cancer cells (IC50: 95.73 ± 9.6 μg/mL) and also had the higher total phenolic (90.5 ± 6.7 μg/mg of gallic acid equivalent (GAE/mg) and flavonoid content (21.8 ± 5.4 μg QE/mg). These results suggest higher antioxidant and cytotoxic activities of leaves extract in comparison with other extracts and these activities could be due to the presence of rich phenolic and flavonoid content. PMID:28255311

  19. Antiproliferative effects of ZnO, ZnO-MTCP, and ZnO-CuMTCP nanoparticles with safe intensity UV and X-ray irradiation

    PubMed Central

    Sadjadpour, Susan; Safarian, Shahrokh; Zargar, Seyed Jalal; Sheibani, Nader

    2016-01-01

    In photodynamic therapy (PDT) of cancer both the light and the photosensitizing agent are normally harmless, but in combination they could result in selective tumor killing. Zinc oxide nanoparticles were synthesized and coated with the amino acid cysteine to provide an adequate arm for conjugation with porphyrin photosensitizers (meso-tetra (4-carboxyphenyl) porphyrin [MTCP] and CuMTCP). Porphyrin-conjugated nanoparticles were characterized by TEM, FTIR, and UV–vis, and fluorescence spectrophotometry. The 3-[4, 5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay was used to measure cell viability in the presence or absence of porphyrin conjugates following UV and X-ray irradiation. The uptake of the porphyrin-conjugated ZnO nanoparticles by cells was detected using fluorescence microscopy. Our results indicated that the survival of T-47D cells was significantly compromised in the presence of ZnO-MTCP-conjugated nanostructures with UV light exposure. Exhibition of cytotoxic activity of ZnO-MTCP for human prostate cancer (Du145) cells occurred at a higher concentration, indicating the more resistant nature of these tumor cells. ZnO-CuMTCP showed milder cytotoxic effects in human breast cancer (T-47D) and no cytotoxic effects in Du145 with UV light exposure, consistent with its lower cytotoxic potency as well as cellular uptake. Surprisingly, none of the ZnO-porphyrin conjugates exhibited cytotoxic effects with X-ray irradiation, whereas ZnO alone exerted cytotoxicity. Thus, ZnO and ZnO-porphyrin nanoparticles with UV or X-ray irradiation may provide a suitable treatment option for various cancers. PMID:25581219

  20. Ascorbic acid protects against colistin sulfate-induced neurotoxicity in PC12 cells.

    PubMed

    Liu, Yang; Dai, Chongshan; Gao, Ruixia; Li, Jichang

    2013-10-01

    This study aimed to examine the protective effect of ascorbic acid against colistin-induced neurotoxicity mediated by oxidative stress, a potential mechanism. An in vitro neurotoxicity model was established with PC12 cells exposed to 125 µg/mL colistin sulfate for 24 h. PC12 cells were treated with colistin (125 µg/mL) in the absence and presence of ascorbic acid (0.1, 1.0 and 10 µM/mL) for 24 h. Both 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) assay were carried out to evaluate cell viability. The levels of intracellular reactive oxygen species (ROS), superoxide dismutase (SOD) and glutathione (GSH) levels were assessed. Moreover, we tested the level of DNA fragmentation, the release of cytochrome-c and the expressions of caspase-9 and -3 mRNA. The results showed that 1 and 10 µM/mL ascorbic acid significantly increased the cell viability and the levels of SOD and GSH (both p<0.05), while 0.1, 1 and 10 µM/mL ascorbic acid significantly decreased the generation of ROS, the release of cytochrome-c, formation of DNA fragmentation and the expressions of caspase-9 and -3 mRNA in colistin-treated PC12 cells, compared with the colistin model group. These results suggest that ascorbic acid could reduce colistin sulfate-induced neurotoxicity through the resistance of oxidative stress and the prevention of apoptosis mediated via mitochondria pathway. They also highlight the potential of coadministering ascorbic acid to widen the therapeutic dose of colistin.

  1. Changes in the functional characteristics of tumor and normal cells after treatment with extracts of white dead-nettle.

    PubMed

    Veleva, Ralitsa; Petkova, Bela; Moskova-Doumanova, Veselina; Doumanov, Jordan; Dimitrova, Milena; Koleva, Petya; Mladenova, Kirilka; Petrova, Svetla; Yordanova, Zhenya; Kapchina-Toteva, Veneta; Topouzova-Hristova, Tanya

    2015-01-02

    Lamium album L. is a perennial herb widely used in folk medicine. It possesses a wide spectrum of therapeutic activities (anti-inflammatory, astringent, antiseptic, antibiotic, antispasmodic, antioxidant and anti-proliferative). Preservation of medicinal plant could be done by in vitro propagation to avoid depletion from their natural habitat. It is important to know whether extracts from L. album plants grown in vitro possess similar properties as extracts from plants grown in vivo. For these reasons, it is important to examine changes in the composition of secondary metabolites during in vitro cultivation of the plant and how they affect the biological activity. We used A549 human cancer cell line and normal kidney epithelial cells MDCKII (Madin-Darby canine kidney cells II) as controls in assessing the anti-cancer effect of plant extracts. To elucidate changes in some key functional characteristics, adhesion test, MTT (3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyl tetrazolium bromide), transepithelial resistance (TER), immunofluorescence staining and trypan blue exclusion test were performed. Methanol and chloroform extracts of in vivo and in vitro propagated plants affected differently cancerous and non-cancerous cells. The most pronounced differences were observed in the morphological analysis and in the cell adhesive properties. We also detected suppressed epithelial transmembrane electrical resistance of MDCK II cells, by treatment with plant extracts, compared to non-treated MDCK II cells. A549 cells did not polarize under the same conditions. Altered organization of actin filaments in both cell types were noticed suggesting that extracts from L. album L. change TER and actin filaments, and somehow may block cell mechanisms, leading to the polarization of MDCK II cells.

  2. Cytotoxic effects of Pinus eldarica essential oil and extracts on HeLa and MCF-7 cell lines.

    PubMed

    Sarvmeili, Najmeh; Jafarian-Dehkordi, Abbas; Zolfaghari, Behzad

    2016-12-01

    Several attempts have so far been made in the search of new anticancer agents of plant origin. Some studies have reported that different species of Pine genus possess cytotoxic activities against various cancer cell lines. In the present study, we evaluated the cytotoxic effects of Pinus eldarica bark and leaf extracts or leaf essential oil on HeLa and MCF-7 tumor cell lines. Hydroalcoholic and phenolic extracts and the essential oil of plant were prepared. Total phenolic contents of the extracts were measured using Folin-Ciocalteu reagent. Essential oil components were determined by gas chromatography-mass spectroscopy (GC-MS). Cytotoxic activity of the extracts and essential oil against HeLa and MCF-7 tumor cell lines were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The polyphenolic content of hydroalcoholic and phenolic extracts of the bark and hydroalcoholic extract of the leaf were 48.31%, 47.2%, and 8.47%, respectively. According to the GC-MS analysis, the major components of the leaf oil of P. eldarica were: β -caryophyllene (14.8%), germacrene D (12.9%), α-terpinenyl acetate (8.15%), α -pinene (5.7%), and -α humulene (5.9%). Bark extracts and leaf essential oil of P. eldarica significantly reduced the viability of both HeLa and MCF-7 cells in a concentration dependent manner. However, leaf extract showed less inhibitory effects against both cell lines. The essential oil of P. eldarica was more cytotoxic than its hydroalcoholic and phenolic extracts. The terpenes and phenolic compounds were probably responsible for cytotoxicity of P. eldarica. Therefore, P. eldarica might have a good potential for active anticancer agents.

  3. Inosine strongly enhances proliferation of human C32 melanoma cells through PLC-PKC-MEK1/2-ERK1/2 and PI3K pathways.

    PubMed

    Soares, Ana Sofia; Costa, Vera Marisa; Diniz, Carmen; Fresco, Paula

    2015-01-01

    Malignant melanoma is the most deadly type of skin cancer. The lack of effective pharmacological approaches for this tumour can be related to the incomplete understanding of the pathophysiological mechanisms involved in melanoma cell proliferation. Adenosine has growth-promoting and growth inhibitory effects on tumour cells. We aimed to investigate effects of adenosine and its metabolic product, inosine, on human C32 melanoma cells and the signalling pathways involved. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction and bromodeoxyuridine (BrdU) proliferation assays were used to evaluate adenosine, adenosine deaminase and inosine effects, in the absence or presence of adenosine receptor (AR), A3 AR and P2Y1 R antagonists and PLC, PKC, MEK1/2 and PI3K inhibitors. ERK1/2 levels were determined using an ELISA kit. Adenosine and inosine levels were quantified using an enzyme-coupled assay. Adenosine caused cell proliferation through AR activation. Adenosine deaminase increased inosine levels (nanomolar concentrations) on the extracellular space, in a time-dependent manner, inducing proliferation through A3 AR activation. Micromolar concentrations of inosine enhanced proliferation through A3 AR activation, causing an increase in ERK1/2 levels, and P2Y1 R activation via ENT-dependent mechanisms. We propose the simultaneous activation of PLC-PKC-MEK1/2-ERK1/2 and PI3K pathways as the main mechanism responsible for the proliferative effect elicited by inosine and its significant role in melanoma cancer progression.

  4. Neuroprotective effects of dimerumic acid and deferricoprogen from Monascus purpureus NTU 568-fermented rice against 6-hydroxydopamine-induced oxidative stress and apoptosis in differentiated pheochromocytoma PC-12 cells.

    PubMed

    Tseng, Wei-Ting; Hsu, Ya-Wen; Pan, Tzu-Ming

    2016-08-01

    Context Oxidative stress plays a key role in neurodegenerative disorders, including Parkinson's disease (PD). Rice fermented with Monascus purpureus Went (Monascaceae) NTU 568 (red mould rice) was found to contain antioxidants, including dimerumic acid (DMA) and deferricoprogen (DFC). Objective The effects of DMA and DFC on 6-hydroxydopamine (6-OHDA)-induced cytotoxicity and potential protective mechanisms in differentiated PC-12 pheochromocytoma cells were investigated. Materials and methods DMA (0-60 μM) or DFC (0-10 μM) was co-treated with 6-OHDA (200 μM, 24 h exposure) in differentiated PC-12 cells. Cell viability and intercellular reactive oxygen species (ROS) were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assays, respectively. Cell apoptosis was determined by DNA fragmentation analysis and propidium iodide staining by flow cytometry. Western blot analysis was used to measure the levels of cell protein expression. Results DMA and DFC significantly increased cell viability to 72% and 81% in 6-OHDA-induced differentiated PC-12 cell cultures, respectively. Furthermore, DMA and DFC reduced 6-OHDA-induced formation of extracellular and intercellular ROS by 25% and 20%, respectively, and decreased NADPH oxidase-2 expression in differentiated PC-12 cells. DMA and DFC inhibited 6-OHDA-induced apoptosis and decreased activation of caspase-3 via regulation of Bcl-2-associated X protein (Bax) and Bcl-2 protein expression in differentiated PC-12 cells. Conclusion DMA and DFC may protect against 6-OHDA toxicity by inhibiting ROS formation and apoptosis. These results showed that the metabolites from M. purpureus NTU 568 fermentation were potential therapeutic agents for PD induced by oxidative damage and should be encouraged for further research.

  5. Comparative proteomic analysis of drug sodium iron chlorophyllin addition to Hep 3B cell line.

    PubMed

    Zhang, Jun; Wang, Wenhai; Yang, Fengying; Zhou, Xinwen; Jin, Hong; Yang, Peng-yuan

    2012-09-21

    The human hepatoma 3B cell line was chosen as an experimental model for in vitro test of drug screening. The drugs included chlorophyllin and its derivatives such as fluo-chlorophyllin, sodium copper chlorophyllin, and sodium iron chlorophyllin. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) method was used in this study to obtain the primary screening results. The results showed that sodium iron chlorophyllin had the best LC(50) value. Proteomic analysis was then performed for further investigation of the effect of sodium iron chlorophyllin addition to the Hep 3B cell line. The proteins identified from a total protein extract of Hep 3B before and after the drug addition were compared by two-dimensional-gel-electrophoresis. Then 32 three-fold differentially expressed proteins were successfully identified by MALDI-TOF-TOF-MS. There are 29 unique proteins among those identified proteins. These proteins include proliferating cell nuclear antigen (PCNA), T-complex protein, heterogeneous nuclear protein, nucleophosmin, heat shock protein A5 (HspA5) and peroxiredoxin. HspA5 is one of the proteins which are involved in protecting cancer cells against stress-induced apoptosis in cultured cells, protecting them against apoptosis through various mechanisms. Peroxiredoxin has anti-oxidant function and is related to cell proliferation, and signal transduction. It can protect the oxidation of other proteins. Peroxiredoxin has a close relationship with cancer and can eventually become a disease biomarker. This might help to develop a novel treatment method for carcinoma cancer.

  6. Effect of Verbascum thapsus ethanol extract on induction of apoptosis in Trichomonas vaginalis in vitro.

    PubMed

    Kashan, Zohreh F; Arbabi, Mohsen; Delavari, Mahdi; Hooshyar, Hossein; Taghizadeh, Mohsen; Joneydy, Zeynab

    2015-01-01

    Trichomoniasis is a sexually transmitted disease (STD) caused by a tiny parasite called Trichomonas vaginalis. Metronidazole is used as routine treatment of disease. Some reports have confirmed the potential carcinogenic and teratogenic effects of this drug on fetus and indication of drug resistance. Verbascum thapsus belongs to the family of Scorphulariaceae. Its antiinflammatory properties, disinfectant and skin healing effects are well known. This plant has been used to treat diarrhea and genitourinary infection in traditional medicine. Effects of different concentrations of the Verbascum thapsus extract were tested on the growth and motility of T. vaginalis trophozoites. To evaluate the toxicity of extract, their effects on mice macrophages were measured by MTT([3-(4,5-dimethyl thiazolyl-2)- 2,5-diphenyle tetrazolium bromide ])assay. In this experimental study the effect of Verbascum thapsus ethanol extract on induction apoptosis in T. vaginalis was determined by Flow Cytometry. Results were analyzed by Flow Jo software and the degree of apoptosis was determined. Toxicity percentage of 25-800 μg/ml concentrations of Verbascum thapsus alcoholic extract for mice macrophages was observed between 0.17-0.25 after 12 hours and they were between 0.25-0.42 and0.45-0.95 after 24 and 48h respectively. IC50 (inhibitory concentration, 50%) of Verbascum thapsus ethanol extract and metronidazole after 24h was 39.17 and 0.0326 μg/ml respectively. Flow cytometry results showed the percent of apoptosis following treatment of trophozoites with different concentrations of Verbascum thapsus ethanol extract (25, 50,100,200,400 μg/ml), were 20.7, 37.04, 47.5, 62.72 and 86.35 respectively, while in control group was 2.9. According to this study, Verbascum thapsus extract induces programmed death in T. vaginalis. It is recommended that Verbascum thapsus extract can be considered as a suitable choice for Medical Studies.

  7. Biological impact of superparamagnetic iron oxide nanoparticles for magnetic particle imaging of head and neck cancer cells

    PubMed Central

    Lindemann, Antje; Lüdtke-Buzug, Kerstin; Fräderich, Bianca M; Gräfe, Ksenija; Pries, Ralph; Wollenberg, Barbara

    2014-01-01

    Background As a tomographic imaging technology, magnetic particle imaging (MPI) allows high spatial resolution and sensitivity, and the possibility to create real-time images by determining the spatial distribution of magnetic particles. To ensure a prospective biosafe application of UL-D (University of Luebeck-Dextran coated superparamagnetic nanoparticles), we evaluated the biocompatibility of superparamagnetic iron oxide nanoparticles (SPIONs), their impact on biological properties, and their cellular uptake using head and neck squamous cancer cells (HNSCCs). Methods SPIONs that met specific MPI requirements were synthesized as tracers. Labeling and uptake efficiency were analyzed by hematoxylin and eosin staining and magnetic particle spectrometry. Flow cytometry, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays, and real-time cell analyzer assays were used to investigate apoptosis, proliferation, and the cytokine response of SPION-labeled cells. The production of reactive oxygen species (ROS) was determined using a fluorescent dye. Experimental results were compared to the contrast agent Resovist®, a standard agent used in MPI. Results UL-D nanoparticles and Resovist particles were taken up in vitro by HNSCCs via unspecific phagocytosis followed by cytosolic accumulation. To evaluate toxicity, flow cytometry analysis was performed; results showed that dose- and time-dependent administration of Resovist induced apoptosis whereas cell viability of UL-D-labeled cells was not altered. We observed decreased cell proliferation in response to increased SPION concentrations. An intracellular production of ROS could not be detected, suggesting that the particles did not cause oxidative stress. Tumor necrosis factor alpha (TNF-α) and interleukins IL-6, IL-8, and IL-1β were measured to distinguish inflammatory responses. Only the primary tumor cell line labeled with >0.5 mM Resovist showed a significant increase in IL-1β secretion

  8. In vitro antioxidant and cytotoxic potential of different parts of Tephrosia purpurea.

    PubMed

    Padmapriya, Ramamoorthy; Ashwini, Sankar; Raveendran, Ramasamy

    2017-02-01

    The antioxidant and cytotoxic properties of four major parts of methanolic extracts of Tephrosia purpurea including leaves, root, stem and seed were investigated and compared. In vitro antioxidant activity of T. purpurea extracts was evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), reducing power assay and antihemolytic assay. In vitro cytotoxic effect of T. purpurea extracts on SW620 colorectal cancer cell line was studied using 3-(4, 5-dimethylthiazolyl -2,5-diphenyl-tetrazolium bromide (MTT) assay. Folin-ciocalteu and aluminium chloride methods were used to determine the total phenolic and flavonoid contents respectively. Among the four extracts studied, leaves extract showed the highest antioxidant activity, DPPH: 186.3 ± 14.0 μg/mL, FRAP: 754.2 ± 50.9 μmol Fe(II)/mg and reducing power activity: 65.7 ± 4.2 μg/mg of quercetin equivalent (QE/mg) and there was no significant difference observed in antihemolytic activity. Leaves extract showed effective cytotoxicity on colorectal cancer cells (IC50: 95.73 ± 9.6 μg/mL) and also had the higher total phenolic (90.5 ± 6.7 μg/mg of gallic acid equivalent (GAE/mg) and flavonoid content (21.8 ± 5.4 μg QE/mg). These results suggest higher antioxidant and cytotoxic activities of leaves extract in comparison with other extracts and these activities could be due to the presence of rich phenolic and flavonoid content.

  9. Early effects of low dose 12C6+ ion or X-ray irradiation on human peripheral blood lymphocytes

    NASA Astrophysics Data System (ADS)

    Chen, Yingtai; Li, Yumin; Zhang, Hong; Xie, Yi; Chen, Xuezhong; Ren, Jinyu; Zhang, Xiaowei; Zhu, Zijiang; Liu, Hongliang; Zhang, Yawei

    2010-04-01

    The aim of this study was to estimate the acute effects of low dose 12C6+ ions or X-ray radiation on human immune function. The human peripheral blood lymphocytes (HPBL) of seven healthy donors were exposed to 0.05 Gy 12C6+ ions or X-ray radiation and cell responses were measured at 24 h after exposure. The cytotoxic activities of HPBL were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT); the percentages of T and NK cells subsets were detected by flow cytometry; mRNA expression of interleukin (IL)-2, tumor necrosis factor (TNF)-α and interferon (IFN)-γ were examined by real time quantitative RT-PCR (qRT-PCR); and these cytokines protein levels in supernatant of cultured cells were assayed by enzyme-linked immunosorbent assays (ELISA). The results showed that the cytotoxic activity of HPBL, mRNA expression of IL-2, IFN-γ and TNF-α in HPBL and their protein levels in supernatant were significantly increased at 24 h after exposure to 0.05 Gy 12C6+ ions radiation and the effects were stronger than observed for X-ray exposure. However, there was no significant change in the percentage of T and NK cells subsets of HPBL. These results suggested that 0.05 Gy high linear energy transfer (LET) 12C6+ radiation was a more effective approach to host immune enhancement than that of low LET X-ray. We conclude that cytokines production might be used as sensitive indicators of acute response to LDI.

  10. Cytotoxic activity of Justicia spicigera is inhibited by bcl-2 proto-oncogene and induces apoptosis in a cell cycle dependent fashion.

    PubMed

    Cáceres-Cortés, J R; Cantú-Garza, F A; Mendoza-Mata, M T; Chavez-González, M A; Ramos-Mandujano, G; Zambrano-Ramírez, I R

    2001-12-01

    Identification of organic compounds from plants is of clinical significance because of the effect that they might have in patients with haematopoietic disorders. We studied the effect of the plant extract Justicia spicigera (Acanthaceae) in different haematopoietic cells: human leukaemic cell lines, umbilical cord blood cells, and mouse bone marrow cells. By examining colony formation and performing the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay it was shown that the plant extract of Justicia spicigera contains cytotoxic factors for leukaemic cells and has no proliferative activity on normal haematopoietic progenitor cells. Our results show that this plant extract induces apoptosis in the human leukaemia cell line TF-1, but not in the bcl-2 transfectant cell line TB-1. Similar results were obtained using a haemopoietic cell line 32D and 32DBcl2. The cultures of umbilical cord blood cells and mouse bone marrow that contain granulocyte-macrophage colony-stimulating factor (GM-CSF) do not proliferate or become terminally differentiated in the presence of the infusion of Justicia spicigera. GM-CSF that acts by abrogating programmed cell death is not sufficient to inhibit the apoptotic stimulus in TF-1 and 32D cells. Moreover mouse fibroblasts (3T3) and two cervical carcinoma cell lines CALO and INBL, undergo apoptosis in the presence of different concentrations of an infusion from the plant. Our data show that there is a strong correlation between the cytotoxic effect and cell proliferation. Together, these results indicate that the plant infusion of Justicia spicigera does not contain any haematopoietic activity, induces apoptosis inhibited by bcl-2 and is linked to cell proliferation.

  11. Synergistic effects of tea polyphenols and ascorbic acid on human lung adenocarcinoma SPC-A-1 cells.

    PubMed

    Li, Wei; Wu, Jian-xiang; Tu, You-ying

    2010-06-01

    Tea polyphenols have been shown to have anticancer activity in many studies. In the present study, we investigated effects of theaflavin-3-3'-digallate (TF(3)), one of the major theaflavin monomers in black tea, in combination with ascorbic acid (AA), a reducing agent, and (-)-epigallocatechin-3-gallate (EGCG), the main polyphenol presented in green tea, in combination with AA on cellular viability and cell cycles of the human lung adenocarcinoma SPC-A-1 cells. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay showed that the 50% inhibition concentrations (IC(50)) of TF(3), EGCG, and AA on SPC-A-1 cells were 4.78, 4.90, and 30.62 micromol/L, respectively. The inhibitory rates of TF(3) combined with AA (TF(3)+AA) and EGCG combined with AA (EGCG+AA) at a molar ratio of 1:6 on SPC-A-1 cells were 54.4% and 45.5%, respectively. Flow cytometry analysis showed that TF(3)+AA and EGCG+AA obviously increased the cell population in the G(0)/G(1) phase of the SPC-A-1 cell cycle from 53.9% to 62.8% and 60.0%, respectively. TF(3)-treated cells exhibited 65.3% of the G(0)/G(1) phase at the concentration of its IC(50). Therefore, TF(3)+AA and EGCG+AA had synergistic inhibition effects on the proliferation of SPC-A-1 cells, and significantly held SPC-A-1 cells in G(0)/G(1) phase. The results suggest that the combination of TF(3) with AA or EGCG with AA enhances their anticancer activity.

  12. Topiramate protects against glutamate excitotoxicity via activating BDNF/TrkB-dependent ERK pathway in rodent hippocampal neurons.

    PubMed

    Mao, Xiao-Yuan; Cao, Yong-Gang; Ji, Zhong; Zhou, Hong-Hao; Liu, Zhao-Qian; Sun, Hong-Li

    2015-07-03

    Topiramate (TPM) was previously found to have neuroprotection against neuronal injury in epileptic and ischemic models. However, whether TPM protects against glutamate-induced excitotoxicity in hippocampal neurons is elusive. Our present work aimed to evaluate the protective effect of TPM against glutamate toxicity in hippocampal neurons and further figure out the potential molecular mechanisms. The in vitro glutamate excitotoxic model was prepared with 125μM glutamate for 20min. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) analysis and Hoechst 33342 staining were conducted to detect neuronal survival. The protein expressions of brain-derived neurotrophic factor (BDNF), TrkB, mitogen-activated protein kinase (MAPK) cascade (including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 MAPK), cyclic AMP response element binding protein (CREB), Bcl-2, Bax and β-actin were detected via Western blot assay. Our results demonstrated that TPM protected hippocampal neurons from glutamate toxicity. Meanwhile, the pretreatment of TPM for 10min significantly prevented the down-regulation of BDNF and the phosphorylation of TrkB. Furthermore, the elevation of phosphorylated EKR expression was significantly inhibited after blockade of TrkB by TrkB IgG, while no alterations of phosphorylated JNK and p38 MAPK were found in the cultured hippocampal neurons. Besides, it was also found that the enhanced phosphorylation of CREB was evidently reversed under excitotoxic conditions after treating with U0126 (the selective inhibitor of ERK). The protein level of Bcl-2 was also observed to be remarkably increased after TPM treatment. In conclusion, these findings implicate that TPM exerts neuroprotective effects against glutamate excitotoxicity in hippocampal neurons and its protection may be modulated through BDNF/TrkB-dependent ERK pathway.

  13. Changes in the functional characteristics of tumor and normal cells after treatment with extracts of white dead-nettle

    PubMed Central

    Veleva, Ralitsa; Petkova, Bela; Moskova-Doumanova, Veselina; Doumanov, Jordan; Dimitrova, Milena; Koleva, Petya; Mladenova, Kirilka; Petrova, Svetla; Yordanova, Zhenya; Kapchina-Toteva, Veneta; Topouzova-Hristova, Tanya

    2015-01-01

    Lamium album L. is a perennial herb widely used in folk medicine. It possesses a wide spectrum of therapeutic activities (anti-inflammatory, astringent, antiseptic, antibiotic, antispasmodic, antioxidant and anti-proliferative). Preservation of medicinal plant could be done by in vitro propagation to avoid depletion from their natural habitat. It is important to know whether extracts from L. album plants grown in vitro possess similar properties as extracts from plants grown in vivo. For these reasons, it is important to examine changes in the composition of secondary metabolites during in vitro cultivation of the plant and how they affect the biological activity. We used A549 human cancer cell line and normal kidney epithelial cells MDCKII (Madin–Darby canine kidney cells II) as controls in assessing the anti-cancer effect of plant extracts. To elucidate changes in some key functional characteristics, adhesion test, MTT (3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyl tetrazolium bromide), transepithelial resistance (TER), immunofluorescence staining and trypan blue exclusion test were performed. Methanol and chloroform extracts of in vivo and in vitro propagated plants affected differently cancerous and non-cancerous cells. The most pronounced differences were observed in the morphological analysis and in the cell adhesive properties. We also detected suppressed epithelial transmembrane electrical resistance of MDCK II cells, by treatment with plant extracts, compared to non-treated MDCK II cells. A549 cells did not polarize under the same conditions. Altered organization of actin filaments in both cell types were noticed suggesting that extracts from L. album L. change TER and actin filaments, and somehow may block cell mechanisms, leading to the polarization of MDCK II cells. PMID:26019631

  14. Effect of bovine lactoferrin on functions of activated feline peripheral blood mononuclear cells during chronic feline immunodeficiency virus infection.

    PubMed

    Kobayashi, Saori; Sato, Reeko; Aoki, Takako; Omoe, Katsuhiko; Inanami, Osamu; Hankanga, Careen; Yamada, Yuichi; Tomizawa, Nobuyuki; Yasuda, Jun; Sasaki, Juso

    2008-05-01

    Feline immunodeficiency virus (FIV) infection is characterized by chronic overactivation of immune and inflammatory system, resulting in anergic state and dysfunction of immune cells. Lactoferrin (LF), a glycoprotein present in exocrine secretions and neutrophils, plays an important role in host defense system. Our previous study showed that oral administration of bovine LF (bLF) suppressed oral inflammation, improved the clinical symptoms and decreased serum gamma-globulin as a marker of inflammation in FIV-infected cats with intractable stomatitis. The anti-inflammatory effect was partly involved in regulation of neutrophil function by bLF. In this study, to clarify the relationship between anti-inflammatory effects of bLF and peripheral blood mononuclear cells (PBMC), we examined the effect of bLF on proliferation, cell cycle progression and cytokine expression in mitogen-activated PBMC. MTT [3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl tetrazolium bromide] assay showed that bLF inhibited the concanavalin A (ConA)-induced cell proliferation in FIV-infected cats with the asymptomatic carrier and AIDS-related complex (ARC) phase. Bovine LF restored ConA-induced cell cycle progression and resulted in suppression of the induced apoptosis in feline PBMC. Real-time RT-PCR showed that bLF suppressed ConA-induced expression of interferon-gamma and interleukin-2 in cells of the ARC group regardless of the time of its addition to the medium. These results suggest the hypothesis that therapy with bLF may have the potential to improve and protect functions of overactivated lymphocytes by modulating the cell proliferation, cell cycle and cytokines expression in cats in terminal stage of FIV infection.

  15. Hydrostatin-TL1, an Anti-Inflammatory Active Peptide from the Venom Gland of Hydrophis cyanocinctus in the South China Sea

    PubMed Central

    Wang, Ningyuan; Huang, Yan; Li, An; Jiang, Hailong; Wang, Jie; Li, Jianzhong; Qiu, Lei; Li, Ka; Lu, Yiming

    2016-01-01

    Tumor necrosis factor (TNF)-α is a pleiotropic cytokine with intense pro-inflammatory and immunomodulatory properties, and anti-TNF-α biologics are effective therapies for various inflammatory diseases such as inflammatory bowel disease (IBD) and sepsis. Snake venom, as a traditional Chinese medicine, has been used in the treatment of inflammatory diseases in China for centuries. In this research, we constructed a venom gland T7 phage display library of the sea snake Hydrophis cyanocinctus to screen bioactive compounds that antagonize TNF-α and identified a novel nine-amino-acid peptide, termed hydrostatin-TL1 (H-TL1). In enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) analyses, H-TL1 inhibited the interaction between TNF-α and TNF receptor 1 (TNFR1). Further, H-TL1 attenuated the cytotoxicity of TNF-α in L929 cells as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. H-TL1 also decreased the mRNA expression of TNF-α/TNFR1 downstream targets and suppressed the phosphorylation of well-characterized proteins of downstream signal transduction pathways in HEK-293 cells. In vivo data demonstrated that H-TL1 protects animals against dextran sodium sulfate (DSS)-induced acute colitis and lipopolysaccharide (LPS)-induced acute shock. Given its significant anti-inflammatory activity in vitro and in vivo, H-TL1 is a potential peptide for the development of new agents to treat TNF-α-associated inflammatory diseases. PMID:27879679

  16. Protective effect of oat bran extracts on human dermal fibroblast injury induced by hydrogen peroxide.

    PubMed

    Feng, Bing; Ma, Lai-ji; Yao, Jin-jing; Fang, Yun; Mei, Yan-ai; Wei, Shao-min

    2013-02-01

    Oat contains different components that possess antioxidant properties; no study to date has addressed the antioxidant effect of the extract of oat bran on the cellular level. Therefore, the present study focuses on the investigation of the protective effect of oat bran extract by enzymatic hydrolysates on human dermal fibroblast injury induced by hydrogen peroxide (H(2)O(2)). Kjeldahl determination, phenol-sulfuric acid method, and high-performance liquid chromatography (HPLC) analysis indicated that the enzymatic products of oat bran contain a protein amount of 71.93%, of which 97.43% are peptides with a molecular range from 438.56 to 1301.01 Da. Assays for 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity indicate that oat peptide-rich extract has a direct and concentration-dependent antioxidant activity. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay and the TdT-mediated digoxigenin-dUTP nick-end labeling (TUNEL) assay for apoptosis showed that administration of H(2)O(2) in human dermal fibroblasts caused cell damage and apoptosis. Pre-incubation of human dermal fibroblasts with the Oatp for 24 h markedly inhibited human dermal fibroblast injury induced by H(2)O(2), but application oat peptides with H(2)O(2) at same time did not. Pre-treatment of human dermal fibroblasts with Oatp significantly reversed the H(2)O(2)-induced decrease of superoxide dismutase (SOD) and the inhibition of malondialdehyde (MDA). The results demonstrate that oat peptides possess antioxidant activity and are effective against H(2)O(2)-induced human dermal fibroblast injury by the enhanced activity of SOD and decrease in MDA level. Our results suggest that oat bran will have the potential to be further explored as an antioxidant functional food in the prevention of aging-related skin injury.

  17. The biocompatibility of titanium in a buffer solution: compared effects of a thin film of TiO2 deposited by MOCVD and of collagen deposited from a gel.

    PubMed

    Popescu, Simona; Demetrescu, Ioana; Sarantopoulos, Christos; Gleizes, Alain N; Iordachescu, Dana

    2007-10-01

    This study aims at evaluating the biocompatibility of titanium surfaces modified according two different ways: (i) deposition of a bio-inert, thin film of rutile TiO(2) by chemical vapour deposition (MOCVD), and (ii) biochemical treatment with collagen gel, in order to obtain a bio-interactive coating. Behind the comparison is the idea that either the bio-inert or the bio-active coating has specific advantages when applied to implant treatment, such as the low price of the collagen treatment for instance. The stability in buffer solution was evaluated by open circuit potential (OCP) for medium time and cyclic voltametry. The OCP stabilized after 5.10(4) min for all the specimens except the collagen treated sample which presented a stable OCP from the first minutes. MOCVD treated samples stabilized to more electropositive values. Numeric results were statistically analysed to obtain the regression equations for long time predictable evolution. The corrosion parameters determined from cyclic curves revealed that the MOCVD treatment is an efficient way to improve corrosion resistance. Human dermal fibroblasts were selected for cell culture tests, taking into account that these cells are present in all bio-interfaces, being the main cellular type of connective tissue. The cells grew on either type of surface without phenotype modification. From the reduction of yellow, water-soluble 3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT cytotoxicity test), MOCVD treated samples offer better viability than mechanically polished Ti and collagen treated samples as well. Cell spreading, as evaluated from microscope images processed by the program Sigma Scan, showed also enhancement upon surface modification. Depending on the experimental conditions, MOCVD deposited TiO(2) exhibits different nanostructures that may influence biological behaviour. The results demonstrate the capacity of integration in simulated physiologic liquids for an implant pretreated by

  18. Lanthanide-Connecting and Lone-Electron-Pair Active Trigonal-Pyramidal-AsO3 Inducing Nanosized Poly(polyoxotungstate) Aggregates and Their Anticancer Activities

    PubMed Central

    Zhao, Jun-Wei; Li, Hai-Lou; Ma, Xing; Xie, Zhigang; Chen, Li-Juan; Zhu, Yongsheng

    2016-01-01

    By virtue of the stereochemical effect of the lone-electron pair located on the trigonal-pyramidal-AsO3 groups and the one-pot self-assembly strategy in the conventional aqueous solution, a series of novel lanthanide-bridging and lone-electron-pair active trigonal-pyramidal-AsO3 inducing nanosized poly(polyoxotungstate) aggregates [H2N(CH3)2]6 Na24H16{[Ln10W16(H2O)30O50](B-α-AsW9O33)8}·97H2O [Ln = EuIII (1), SmIII (2), GdIII (3), TbIII (4), DyIII (5), HoIII (6), ErIII (7), TmIII (8)] were prepared and further characterized by elemental analyses, IR spectra, UV spectra, thermogravimetric (TG) analyses and single-crystal X-ray diffraction. The most remarkable structural feature is that the polyanionic skeleton of {[Ln10W16(H2O)30O50](B-α-AsW9O33)8}46− is constructed from eight trivacant Keggin [B-α-AsW9O33]9− fragments through ten Ln centers and sixteen bridging W atoms in the participation of fifty extraneous oxygen atoms. Notably, 4 and 8 can be stable in the aqueous solution not only for eight days but also in the range of pH = 3.9–7.5. Moreover, the cytotoxicity tests of 4 and 8 toward human cervical cancer (HeLa) cells, human breast cancer (MCF–7) cells and mouse fibroblast (L929) cells were performed by the 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and the cell apoptosis processes were characterized by calcein AM/PI staining experiments, annexin V-FITC/PI staining experiments and morphological changes. PMID:27193961

  19. Cytotoxicity of Various Endodontic Materials on Stem Cells of Human Apical Papilla

    PubMed Central

    Saberi, Eshagh Ali; Karkehabadi, Hamed; Mollashahi, Narges Farhad

    2016-01-01

    Introduction: This in vitro study assessed and compared the cytotoxicity of mineral trioxide aggregate (MTA), calcium-enriched mixture (CEM) cement, Biodentine (BD) and octacalcium phosphate (OCP) on stem cells of the human apical papilla (SCAP). Methods and Materials: SCAPs were isolated from two semi-impacted third molars. The cells were cultured in wells of an insert 24-well plate and were then incubated. The plates were then removed from the incubator and randomly divided into four experimental groups that were exposed to 1-mm discs of set MTA, CEM, BD or OCP, and one untreated control group. After 24, 48 and 168 h, the plates were removed from the incubator and 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) solution was added to each well. Data were analyzed at different time points using the repeated measures ANOVA followed by Bonferroni test and the level of significance was set at 0.05. Results: Cytotoxicity of the four materials was not significantly different from that of the control group at 24, 48 and 168 h (P>0.05). Two-by-two comparison revealed that cytotoxicity of MTA and CEM cement was significantly different from each other at 168 h (P<0.05) although the cytotoxicity of CEM was less than MTA. Cytotoxicity of OCP and MTA was also significantly different from each other at 48 h and OCP had more favorable biocompatibility than MTA (P<0.05). Conclusion: CEM, OCP, BD and MTA showed acceptable biocompatibility when exposed to SCAP. Over time, CEM showed the least cytotoxicity among the materials under study. PMID:26843872

  20. Expression, characterization, and evaluation of a RANK-binding single chain fraction variable: an osteoclast targeting drug delivery strategy.

    PubMed

    Newa, Madhuri; Lam, Michael; Bhandari, Krishna Hari; Xu, Biwen; Doschak, Michael R

    2014-01-06

    A single chain Fraction variable (scFv) employs antibody-like target recognition specificity. Osteoclasts, responsible for bone resorption, express Receptor Activator of Nuclear factor Kappa B (RANK) receptors. This study aimed to express, characterize, and evaluate scFv against RANK receptors that may serve as a platform to target osteoclasts. Using phage display technology, scFv against RANK receptor was expressed and characterized by DNA sequencing, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), matrix-assisted laser desorption-ionization time-of-flight (MALDI TOF), enzyme-linked immunosorbent assay (ELISA), Western blot, and immunocytochemistry. The potential for cytotoxicity was evaluated using an MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay, and its cross reactivity was evaluated using ELISA. Osteoclast-like cells were generated from RAW 264.7 cells, and the osteoclast targeting ability of scFv was evaluated using immunocytochemistry. ScFv's antiresorptive efficacy was studied using a tartrate-resistant acid phosphatase (TRAP) assay and resorption assay. Anti-RANK scFv was successfully expressed and characterized. No cross reactivity with other tumor necrosis factor receptor (TNFR) members and no cytotoxic effect on a non-RANK bearing cell line were observed. It showed specificity toward a RANK receptor and an inhibitory effect on osteoclast activity. With the increase in development trends for biologics as therapeutics and growing knowledge on the importance of osteoclast targeted therapy, this study may provide a drug delivery strategy to target osteoclasts, thereby leading to a promising therapy for resorptive bone diseases.

  1. Surfactant, but not the size of solid lipid nanoparticles (SLN) influences viability and cytokine production of macrophages.

    PubMed

    Schöler, N; Olbrich, C; Tabatt, K; Müller, R H; Hahn, H; Liesenfeld, O

    2001-06-19

    After intravenous (i.v.) injection, solid lipid nanoparticles (SLN) interact with mononuclear cells. Murine peritoneal macrophages were incubated with SLN formulations consisting of Dynasan 114 coated with different surfactants. The present study was performed to examine the impact of surfactants, which are important surface defining components of SLN, on viability and cytokine production by macrophages. Cytotoxicity, as assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) test, was strongly influenced by the surfactant used being marked with cetylpyridinium chloride- (CPC-) coated SLN at a concentration of 0.001% and further increased at SLN concentrations of 0.01 and 0.1%. All other SLN formulations -- containing Poloxamine 908 (P908), Poloxamer 407 (P407), Poloxamer 188 (P188), Solutol HS15 (HS15), Tween 80 (T80), Lipoid S75 (S75), sodium cholate (SC), or sodium dodecylsulfate (SDS) -- when used at the same concentrations reduced cell viability only slightly. None of the SLN formulations tested induced cytokine production but a concentration-dependent decrease of IL-6 production was observed, which appeared to be associated with cytotoxic effects. IL-12 and TNF-alpha were detected neither in supernatants of macrophages treated with SLN at any concentration nor in those of untreated cells. In contrast to the type of surfactant, the size of SLN was found neither to affect cytotoxicity of SLN nor to result in induction or digression of cytokine production by macrophages. In conclusion, testing the effects of surfactants on SLN on activity of macrophages is a prerequisite prior to in vivo use of SLN.

  2. Effect of cinnamon oil on icaA expression and biofilm formation by Staphylococcus epidermidis.

    PubMed

    Nuryastuti, Titik; van der Mei, Henny C; Busscher, Henk J; Iravati, Susi; Aman, Abu T; Krom, Bastiaan P

    2009-11-01

    Staphylococcus epidermidis is notorious for its biofilm formation on medical devices, and novel approaches to prevent and kill S. epidermidis biofilms are desired. In this study, the effect of cinnamon oil on planktonic and biofilm cultures of clinical S. epidermidis isolates was evaluated. Initially, susceptibility to cinnamon oil in planktonic cultures was compared to the commonly used antimicrobial agents chlorhexidine, triclosan, and gentamicin. The MIC of cinnamon oil, defined as the lowest concentration able to inhibit visible microbial growth, and the minimal bactericidal concentration, the lowest concentration required to kill 99.9% of the bacteria, were determined using the broth microdilution method and plating on agar. A checkerboard assay was used to evaluate the possible synergy between cinnamon oil and the other antimicrobial agents. The effect of cinnamon oil on biofilm growth was studied in 96-well plates and with confocal laser-scanning microscopy (CLSM). Biofilm susceptibility was determined using a metabolic 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Real-time PCR analysis was performed to determine the effect of sub-MIC concentrations of cinnamon oil on expression of the biofilm-related gene, icaA. Cinnamon oil showed antimicrobial activity against both planktonic and biofilm cultures of clinical S. epidermidis strains. There was only a small difference between planktonic and biofilm MICs, ranging from 0.5 to 1% and 1 to 2%, respectively. CLSM images indicated that cinnamon oil is able to detach and kill existing biofilms. Thus, cinnamon oil is an effective antimicrobial agent to combat S. epidermidis biofilms.

  3. Ameliorating effects of aged garlic extracts against Aβ-induced neurotoxicity and cognitive impairment

    PubMed Central

    2013-01-01

    Background In vitro antioxidant activities and neuron-like PC12 cell protective effects of solvent fractions from aged garlic extracts were investigated to evaluate their anti-amnesic functions. Ethyl acetate fractions of aged garlic had higher total phenolics than other fractions. Methods Antioxidant activities of ethyl acetate fractions from aged garlic were examined using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) and malondialdehyde (MDA) inhibitory effect using mouse whole brain homogenates. Levels of cellular oxidative stress as reactive oxygen species (ROS) accumulation were measured using 2',7'-dichlorofluorescein diacetate (DCF-DA). PC12 cell viability was investigated by 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and lactate dehydtrogenase (LDH) assay. The learning and memory impairment in institute of cancer research (ICR) mice was induced by neurotoxic amyloid beta protein (Aβ) to investigate in vivo anti-amnesic effects of aged garlic extracts by using Y-maze and passive avoidance tests. Results We discovered that ethyl acetate fractions showed the highest ABTS radical scavenging activity and MDA inhibitory effect. Intracellular ROS accumulation resulting from Aβ treatment in PC12 cells was significantly reduced when ethyl acetate fractions were presented in the medium compare to PC12 cells which was only treated with Aβ only. Ethyl acetate fractions from aged garlic extracts showed protection against Aβ-induced neurotoxicity. Pre-administration with aged garlic extracts attenuated Aβ-induced learning and memory deficits in both in vivo tests. Conclusions Our findings suggest that aged garlic extracts with antioxidant activities may improve cognitive impairment against Aβ-induced neuronal deficit, and possess a wide range of beneficial activities for neurodegenerative disorders, notably Alzheimer's disease (AD). PMID:24134394

  4. Assessing cytotoxicity of boron nitride nanotubes: Interference with the MTT assay

    SciTech Connect

    Ciofani, Gianni; Danti, Serena; D'Alessandro, Delfo; Moscato, Stefania; Menciassi, Arianna

    2010-04-02

    Thanks to a non-covalent wrapping with glycol-chitosan, highly biocompatible and highly concentrated dispersions of boron nitride nanotubes were obtained and tested on human neuroblastoma cells. A systematic investigation of the cytotoxicity of these nanovectors with several complementary qualitative and quantitative assays allowed a strong interference with the MTT metabolic assay to be highlighted, similar to a phenomenon already observed for carbon nanotubes, that would wrongly suggest toxicity of boron nitride nanotubes. These results confirm the high complexity of these new nanomaterials, and the needing of extensive investigations on their exciting potential applications in the biomedical field.

  5. The Multidisciplinary Translational Team (MTT) Model for Training and Development of Translational Research Investigators

    PubMed Central

    Ameredes, Bill T.; Hellmich, Mark R.; Cestone, Christina M.; Wooten, Kevin C.; Ottenbacher, Kenneth J.; Chonmaitree, Tasnee; Anderson, Karl E.; Brasier, Allan R.

    2015-01-01

    Multi-institutional research collaborations now form the most rapid and productive project execution structures in the health sciences. Effective adoption of a multidisciplinary team research approach is widely accepted as one mechanism enabling rapid translation of new discoveries into interventions in human health. Although the impact of successful team-based approaches facilitating innovation has been well-documented, its utility for training a new generation of scientists has not been thoroughly investigated. We describe the characteristics of how multidisciplinary translational teams (MTTs) promote career development of translational research scholars through competency building, inter-professional integration, and team-based mentoring approaches. Exploratory longitudinal and outcome assessments from our experience show that MTT membership had a positive effect on the development of translational research competencies, as determined by a self-report survey of 32 scholars. We also observed that all trainees produced a large number of collaborative publications that appeared to be associated with their CTSA association and participation with MTTs. We conclude that the MTT model provides a unique training environment for translational and team-based learning activities, for investigators at early stages of career development. PMID:26010046

  6. The Multidisciplinary Translational Team (MTT) Model for Training and Development of Translational Research Investigators.

    PubMed

    Ameredes, Bill T; Hellmich, Mark R; Cestone, Christina M; Wooten, Kevin C; Ottenbacher, Kenneth J; Chonmaitree, Tasnee; Anderson, Karl E; Brasier, Allan R

    2015-10-01

    Multiinstitutional research collaborations now form the most rapid and productive project execution structures in the health sciences. Effective adoption of a multidisciplinary team research approach is widely accepted as one mechanism enabling rapid translation of new discoveries into interventions in human health. Although the impact of successful team-based approaches facilitating innovation has been well-documented, its utility for training a new generation of scientists has not been thoroughly investigated. We describe the characteristics of how multidisciplinary translational teams (MTTs) promote career development of translational research scholars through competency building, interprofessional integration, and team-based mentoring approaches. Exploratory longitudinal and outcome assessments from our experience show that MTT membership had a positive effect on the development of translational research competencies, as determined by a self-report survey of 32 scholars. We also observed that all trainees produced a large number of collaborative publications that appeared to be associated with their CTSA association and participation with MTTs. We conclude that the MTT model provides a unique training environment for translational and team-based learning activities, for investigators at early stages of career development.

  7. METHYL BROMIDE ALTERNATIVES FOR CALIFORNIA STRAWBERRY NURSERIES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects of methyl bromide (MB) alternative fumigants on soil pests, plant productivity in nursery and fruiting fields, as well as production costs, were evaluated in California strawberry nurseries by an interdisciplinary team. Our trials followed nursery stock through low and high elevation ph...

  8. Methyl bromide emissions from tarped fields

    SciTech Connect

    Cicerone, R.J.; Williams, J.; Wang, N.Y.

    1995-12-31

    Once in the stratosphere, bromine atoms can destroy ozone effectively. Because of this potential effect, certain organobromine compounds including methyl bromide (MeBr) are being controlled or eliminated by national and international regulations. It would be valuable to determine the fraction of MeBr used in soil fumigations that subsequently enters the atmosphere to better assess the need for, and value of, strong regulations. We have designed and conducted several experiments accompanying field fumigations with MeBr/chloropicrin mixtures. In each of three field-fumigation experiments new Irvine, CA in which the fumigated field was covered immediately with plastic tarping, we have deployed static flux chambers on top of the tarping and measured escape fluxes of MeBr. After tarp removal, the same chambers were replaced on the bare soil to continue the measurements. We have also measured soil bromide contents before and after the fumigation. One experiment yielded an escape fraction of 80 to 87% (with 19% remaining as bromide) while the other two experiments yielded escape fractions of 30 to 35% (with about 70% remaining as bromide). This paper will summarize stratospheric bromine chemistry, describe the field experiments and discuss factors that influence emissions, including soil pH, moisture and organic content and injection technique. We acknowledge TriCal, Inc. for many helpful discussions and for professional field applications of MeBr.

  9. Cherry angiomas associated with exposure to bromides.

    PubMed

    Cohen, A D; Cagnano, E; Vardy, D A

    2001-01-01

    Cherry angiomas are the most common vascular proliferation; however, little is known about the pathogenesis and etiology of these lesions. We present two laboratory technicians who were exposed to brominated compounds for prolonged periods and who developed multiple cherry angiomas on the trunk and extremities. We suggest that the association between exposure to bromides and cherry angiomas should be investigated by a controlled study.

  10. Methyl Bromide Investigation Expands to Puerto Rico

    EPA Pesticide Factsheets

    (New York, N.Y.) The U.S. Environmental Protection Agency, working with the Virgin Islands government, is investigating the March 2015 incident involving the use of a pesticide containing methyl bromide to fumigate a residence at the Sirenusa apartments on

  11. Flow injection determination of bromide ion in a developer using bromide ion-selective electrode detector.

    PubMed

    Masadome, T; Asano, Y; Nakamura, T

    1999-10-01

    A potentiometric flow injection determination method for bromide ion in a developer was proposed, by utilizing a flow-through type bromide ion-selective electrode detector. The sensing membrane of the electrode was Ag(2)S-AgBr membrane. The response of the electrode detector as a peak-shape signal was obtained for injected bromide ion in a developer. A linear relationship was found to exist between peak height and the concentration of the bromide ion in a developer in a concentration range from 1.0x10(-3) to 1.0x10(-2) mol l(-1). The relative standard deviation for 10 injections of a 6x10(-3) mol l(-1) bromide ion in a developer was 1.3% and the sampling rate was ca 17-20 samples h(-1). The present method was free from the interference of an organic reducing reagent, an organic substance in a developer sample solution for the determination of bromide ion in a developer.

  12. Effects of several salt marsh plants on mouse spleen and thymus cell proliferation using mtt assay

    NASA Astrophysics Data System (ADS)

    Seo, Youngwan; Lee, Hee-Jung; Kim, You Ah; Youn, Hyun Joo; Lee, Burm-Jong

    2005-12-01

    In the present study, we have tested the effects of 21 salt marsh plants on cell proliferation of mouse immune cells (spleen and thymus) using MTT assay in culture. The methanolic extracts of six salt marsh plants ( Rosa rugosa, Ixeris tamagawaensis, Artemisia capillaris, Tetragonia tetragonoides, Erigeron annus, and Glehnia littoralis) showed very powerful suppressive effects of mouse immune cell death and significant activities of cell proliferation in vitro. Especially, the methanolic extract of Rosa rugosa was found to have fifteen times compared to the control treatment, demonstrating that Rosa rugosa may have a potent stimulation effect on immune cell proliferation. These results suggest that several salt marsh plants including Rosa rugosa could be useful for further study as an immunomodulating agent.

  13. Measurements of atmospheric methyl bromide and bromoform

    SciTech Connect

    Cicerone, R.J.; Heidt, L.E.; Pollock, W.H.

    1988-04-20

    We have measured gaseous methyl bromide (CH/sub 3/Br) and bromoform (CHBr/sub 3/) in air samples that were gathered approximately weekly from five ground-level sites: Point Barrow, Alaska; Mauna Loa Observatory and Cape Kumukahi, Hawaii; Matatula, Samoa; and Kaitorete Spit, New Zealand. Approximately 750 samples have been analyzed for CH/sub 3/Br between January 1985 and October 1987 and 990 samples have been analyzed for CHBr/sub 3/ between early 1984 and September 1987, all by gas chromatography/mass spectroscopy. Methyl bromide concentrations are typically 10--11 parts per trillion (ppt) by volume; there are no clear indications of temporal increases. Bromoform concentrations are typically 2--3 ppt, but large seasonal variations are seen at Point Barrow. copyright American Geophysical Union 1988

  14. Thallium bromide iodide crystal acoustic anisotropy examination.

    PubMed

    Mantsevich, S N

    2017-03-01

    Thallium bromide iodide crystal also known as KRS-5 is the well known material used in far infrared radiation applications for optical windows and lenses fabrication. The main advantage of this material is the transparency in wide band of wavelengths from 0.53 to 50μm. Despite such advantages as transparency and large acousto-optic figure of merit values, KRS-5 is rarely used in acousto-optics. Nevertheless this material seems to be promising for far infrared acousto-optic applications. The acoustic and acousto-optic properties of KRS-5 needed for the full use in optoelectronics are not well understood to date. In this paper the detailed examination of thallium bromide iodide crystal acoustic properties is presented.

  15. Data for methyl bromide decon testing

    EPA Pesticide Factsheets

    Spreadsheets containing data for recovery of spores from different materials. Data on the fumigation parameters are also included.This dataset is associated with the following publication:Wood , J., M. Wendling, W. Richter, A. Lastivka, and L. Mickelsen. Evaluation of the Efficacy of Methyl Bromide in the Decontamination of Building and Interior Materials Contaminated with Bacillus anthracis Spores. APPLIED AND ENVIRONMENTAL MICROBIOLOGY. American Society for Microbiology, Washington, DC, USA, 1-28, (2016).

  16. Ipratropium bromide in children with asthma.

    PubMed Central

    Mann, N P; Hiller, E J

    1982-01-01

    Eighteen children between 6 and 14 years of age with perennial asthma were studied over two four-week treatment periods. Ipratropium bromide, given in addition to their current treatment, was compared with placebo using a double-blind crossover technique. The period of treatment with ipratropium was associated with a significant reduction in symptoms during both day and night and significantly higher morning peak expiratory flow rates. PMID:6461943

  17. Methyl bromide users search for science

    SciTech Connect

    Winegar, E.D.

    1995-01-01

    Workers, neighbors and the ozone are protected by regulation from this chemical, but those needing it complain that a solid foundation is lacking for the rules. Although not yet featured on {open_quotes}60 Minutes,{close_quotes} the pesticide methyl bromide is gaining widespread attention because of its central position in debates about worker health and safety, environmental toxics exposure and global ozone depletion.

  18. [Research advances in methyl bromide in the ocean].

    PubMed

    Du, Hui-na; Xie, Wen-xia; Cui, Yu-qian; Chen, Jian-lei; Ye, Si-yuan

    2014-12-01

    Methyl bromide is an important atmospheric trace gas, which plays significant roles in the global warming and atmospheric chemistry. The ocean plays important and complex roles in the global biogeochemical cycles of methyl bromide, not only the source of atmospheric methyl bromide, but also the sink. Therefore, developing the chemical research of the soluble methyl bromide in the ocean, will not only have a certain guiding significance to the atmospheric ozone layer protection, but also provide a theoretical basis for estimating methyl bromide's contribution to the global environmental change on global scale. This paper reviewed the research advances on methyl bromide in the ocean, from the aspects of the biogeochemical cycle of methyl bromide in the ocean, the analysis and determination method, the concentration distribution, the sea-to-air flux and its sources and sinks in the atmosphere. Some deficiencies in the current studies were put forward, and the directions of the future studies were prospected.

  19. Methyl bromide volatility measurements from treated fields

    SciTech Connect

    Majewski, M.S.; Woodrow, J.E.; Seiber, J.N. |

    1995-12-31

    Methyl bromide is used as an agricultural soil fumigant and concern is growing over the role it may play in the depletion of stratospheric ozone. Methyl bromide is applied using various techniques and little is known about how much of the applied fumigant volatilizes into the atmosphere after application. The post-application volatilization losses of methyl bromide from two fields using different application practices were measured using an aerodynamic-gradient technique. One field was covered with a high-barrier plastic film tarp during application and the other was left uncovered, but the furrows made by the injection shanks were bedded over. The cumulative volatilization losses from the tarped field were 22% of the nominal application within the first 5 days of the experiment and about 32% of the nominal application within 9 days including the one day after the tarp was removed on day 8. The nontarped field lost 89%of the nominal application by volatilization in 5 days. The error associated, with each flux measurement, as well as variations in daily flux losses with differing sampling period lengths show the degree of variability inherent in this type of study.

  20. Methyl bromide fate in fumigated soils

    SciTech Connect

    Anderson, T.A.; Rice, P.J.; Cink, J.H.

    1995-12-31

    Although widespread use of methyl bromide (MeBr) as a sail and structure fumigant has previously been recognized as a potential significant source of atmospheric MeBr, losses have not been well quantified. Our research indicates that, in laboratory studies, MeBr is volatilized rapidly from fumigated soils and that volatility increases with temperature (35{degrees}C > 25{degrees}C and 15{degrees}C) and moisture (0.03 bar and 0.3 bar > 1 bar > 3 bar). Degradation of MeBr in soil, as indicated by production of bromide ion, was also directly related to temperature and moisture. Most of the soil degradation of MeBr in these studies appears to be abiotic based on the observation of toxicity (reduced microbial respiration) in fumigated soils. We also determined the transformation and movement of MeBr in undisturbed soil columns. These studies also indicated that MeBT volatilizes rapidly (> 50% in 48 h) from soil. In addition, MeBr was not detected in the leachate from the soil columns, however, bromide ion was detected at levels above background 48 h after fumigation and peaked at 5 weeks.

  1. The neurological effects of methyl bromide intoxication.

    PubMed

    de Souza, Aaron; Narvencar, Kedareshwar P S; Sindhoora, K V

    2013-12-15

    Used primarily as a fumigant or as a substrate in chemical processes, methyl bromide is a highly toxic gas. The gas is usually absorbed by inhalation and effects on the lungs, gastrointestinal tract, skin, and brain are seen. Numerous instances of acute and chronic neurologic injury have been reported: acute poisoning results in seizures, myoclonus, ataxia or cerebral oedema beginning as early as 30 min after exposure while subacute or chronic intoxication presents with diverse slowly progressive neurological and neurobehavioral symptoms. Serum bromide levels may be elevated, but often return rapidly to normal. Electroencephalography may show frontally-predominant slow waves or polyspikes with following slow wave, and MRI reveals characteristic involvement in the dentate nucleus of the cerebellum, the brainstem, and the splenium of the corpus callosum. Symmetric and selective lesions in characteristic sites are observed on imaging and on histopathological examination. These are likely produced by methylation of intracellular lipids, protein and glutathione; production of toxic metabolites; defective neurotransmitter function; and abnormal oxidative phosphorylation. This article reviews the toxic effects of this gas, the pathophysiology and symptoms of its effects on the nervous system, and characteristic findings on MRI; and presents an illustrative case of methyl bromide intoxication due to exposure at a factory producing the compound commercially.

  2. Ipratropium bromide-mediated myocardial injury in in vitro models of myocardial ischaemia/reperfusion.

    PubMed

    Harvey, Kate L; Hussain, Afthab; Maddock, Helen L

    2014-04-01

    Ipratropium bromide, a nonselective muscarinic antagonist, is widely prescribed for the treatment of chronic obstructive pulmonary disease (COPD). Analyses of COPD patients, with underlying ischaemic heart disease, receiving anticholinergics, have indicated increased risk of severity and occurrence of cardiovascular events (including myocardial infarction). The present study explored whether ipratropium bromide induces myocardial injury in nonclinical models of simulated myocardial ischaemia/reperfusion injury. Adult Sprague Dawley rat hearts/primary ventricular myocytes were exposed to simulated ischaemia/hypoxia prior to administration of ipratropium at the onset of reperfusion/reoxygenation. Infarct to risk ratio and cell viability was measured via triphenyl tetrazolium chloride staining and thiazolyl blue tetrazolium bromide (MTT) assay. The involvement of apoptosis and necrosis was evaluated by flow cytometry. Mitochondrial-associated responses were detected by tetramethylrhodamine methyl ester fluorescence and myocyte contracture. Ipratropium (1 × 10⁻¹¹ M - 1 × 10⁻⁴ M) significantly increased infarct/risk ratio and decreased cell viability in a dose-dependent manner. Increased levels of necrosis and apoptosis were observed via flow cytometry, accompanied by increased levels of cleaved caspase-3 following ipratropium treatment. Levels of endogenous myocardial acetylcholine were verified via use of an acetylcholine assay. In these experimental models, exogenous acetylcholine (1 × 10⁻⁷ M) showed protective properties, when administered alone, as well as abrogating the exacerbation of myocardial injury during ischaemia/reperfusion following ipratropium coadministration. In parallel experiments, under conditions of myocardial ischaemia/reperfusion, a similar injury was observed following atropine (1 × 10⁻⁷ M) administration. These data demonstrate for the first time in a nonclinical setting that ipratropium exacerbates ischaemia

  3. Ruthenium(II) sulfoxide-maltolato and -nitroimidazole complexes: synthesis and MTT assay.

    PubMed

    Wu, Adam; Kennedy, David C; Patrick, Brian O; James, Brian R

    2003-11-17

    Ru(II) sulfoxide-maltolato complexes, Ru(ma)(2)(L)(2) (L = DMSO (1a) and TMSO (1b) or L(2) = BESE (1c)), were synthesized, as well as the analogous ethylmaltolato derivatives, Ru(etma)(2)(L)(2) (2a-c) (ma = 3-hydroxy-2-methylpyran-4-onate, etma = 2-ethyl-3-hydroxypyran-4-onate, TMSO = tetramethylene sulfoxide, BESE = 1,2-bis(ethylsulfinyl)ethane). A Ru(II) bidentate sulfoxide-metronidazole complex, RuCl(2)(BESE)(metro)(2) (3), was also synthesized (metro = metronidazole = 2-methyl-5-nitroimidazole-1-ethanol). The complexes were characterized generally by (1)H NMR, UV-vis, and IR spectroscopies, as well as MS, elemental analysis, solution conductivity, and cyclic voltammetry. The molecular structures of Ru(ma)(2)(S,R-BESE) (1c) and trans-RuCl(2)(R,R-BESE)(metro)(2) (3) were determined by X-ray crystallography. All sulfoxide ligands are S-bonded. The complexes were tested against human breast cancer cells (MDA-MB-435S) using an in vitro MTT assay, a colorimetric determination of cell viability: 2a,b exhibit the lowest IC(50) values of 190 +/- 10 and 220 +/- 10 microM, respectively. Cisplatin exhibits an IC(50) value of 30 +/- 5 microM.

  4. Europium-doped barium bromide iodide

    SciTech Connect

    Gundiah, Gautam; Hanrahan, Stephen M.; Hollander, Fredrick J.; Bourret-Courchesne, Edith D.

    2009-10-21

    Single crystals of Ba0.96Eu0.04BrI (barium europium bromide iodide) were grown by the Bridgman technique. The title compound adopts the ordered PbCl2 structure [Braekken (1932). Z. Kristallogr. 83, 222-282]. All atoms occupy the fourfold special positions (4c, site symmetry m) of the space group Pnma with a statistical distribution of Ba and Eu. They lie on the mirror planes, perpendicular to the b axis at y = +-0.25. Each cation is coordinated by nine anions in a tricapped trigonal prismatic arrangement.

  5. Advanced hydrogen electrode for hydrogen-bromide battery

    NASA Technical Reports Server (NTRS)

    Kosek, Jack A.; Laconti, Anthony B.

    1987-01-01

    Binary platinum alloys are being developed as hydrogen electrocatalysts for use in a hydrogen bromide battery system. These alloys were varied in terms of alloy component mole ratio and heat treatment temperature. Electrocatalyst evaluation, performed in the absence and presence of bromide ion, includes floating half cell polarization studies, electrochemical surface area measurements, X ray diffraction analysis, scanning electron microscopy analysis and corrosion measurements. Results obtained to date indicate a platinum rich alloy has the best tolerance to bromide ion poisoning.

  6. [Spectrophotometer detection of benzalkonium bromide concentration in seawater].

    PubMed

    Pan, Jianyu; Yin, Pinghe; Zhao, Ling; Qi, Yuzao; Xie, Longchu

    2003-07-01

    Benzalkonium bromide is a high-efficiency algaecide. Its concentration in seawater was measured by the method of spectrophotometer. The results indicated that the deposition appeared if the concentration of benzalkonium bromide in seawater was more than 50 mg.L-1, and affected the detection of benzalkonium bromide. But, there was a good linear relationship between concentration and absorbance when the concentration was lower than 50 mg.L-1 (R2 = 0.9996). Therefore, spectrophotometer could be used to detect benzalkonium bromide in seawater.

  7. Comparison of the 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide tube method with the conventional method and real-time polymerase chain reaction for the detection of rifampicin resistance in Mycobacterium tuberculosis.

    PubMed

    Raut, U; Rantai, S; Narang, P; Chauhan, D S; Chahar, M; Narang, R; Mendiratta, D K

    2012-01-01

    Colorimetric methods are cheap, reproducible, and rapid methods of detecting drug resistance in Mycobacterium tuberculosis. The MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) method is one such technique that has been established in our laboratory to detect rifampicin resistance. The present study compared the results of the MTT method with those of the proportion method and real-time polymerase chain reaction (RTPCR) in order to establish sensitivity and specificity of MTT. The mutations for rifampicin resistance occur in rpoB gene, and the commonest reported are in codons 526 and 531. Therefore, RTPCR was targeted at these two codons. The concordance of MTT with the proportion method and RTPCR was 94 and 72.77%, respectively, and that of RTPCR with the proportion method was 77.77%. While the study confirmed that the MTT method is a good method for detecting rifampicin resistance, it also brought out the fact that RTPCR when targeted for limited mutations is not a good tool. Either the genotypic method used should target the total 81-bp rpoB genome or methods such as DNA sequencing should be used. For resource-constraint laboratories, the MTT method can be considered as a better choice.

  8. An Ill Wind: Methyl Bromide Use Near California Schools, 1998.

    ERIC Educational Resources Information Center

    Ross, Zev; Walker, Bill

    A California study investigates the use of the toxic pesticide methyl bromide near the state's public schools, explains why proposed safety rules have failed to protect children and others from exposure, and examines regions at particular exposure risk. Study results show an increasing exposure to methyl bromide near schools already at risk while…

  9. 40 CFR 721.10393 - Sodium bromide MDA complex (generic).

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 31 2014-07-01 2014-07-01 false Sodium bromide MDA complex (generic... Specific Chemical Substances § 721.10393 Sodium bromide MDA complex (generic). (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified generically as sodium...

  10. Sir Charles Locock and potassium bromide.

    PubMed

    Eadie, M J

    2012-01-01

    On 12 May 1857, Edward Sieveking read a paper on epilepsy to the Royal Medical and Chirurgical Society in London. During the discussion that followed Sir Charles Locock, obstetrician to Queen Victoria, was reported to have commented that during the past 14 months he had used potassium bromide to successfully stop epileptic seizures in all but one of 14 or 15 women with 'hysterical' or catamenial epilepsy. This report of Locock's comment has generally given him credit for introducing the first reasonably effective antiepileptic drug into medical practice. However examination of the original reports raises questions as to how soundly based the accounts of Locock's comments were. Subsequently, others using the drug to treat epilepsy failed to obtain the degree of benefit that the reports of Locock's comments would have led them to expect. The drug might not have come into more widespread use as a result, had not Samuel Wilks provided good, independent evidence for the drug's antiepileptic efficacy in 1861.

  11. Single ion dynamics in molten sodium bromide

    SciTech Connect

    Alcaraz, O.; Trullas, J.; Demmel, F.

    2014-12-28

    We present a study on the single ion dynamics in the molten alkali halide NaBr. Quasielastic neutron scattering was employed to extract the self-diffusion coefficient of the sodium ions at three temperatures. Molecular dynamics simulations using rigid and polarizable ion models have been performed in parallel to extract the sodium and bromide single dynamics and ionic conductivities. Two methods have been employed to derive the ion diffusion, calculating the mean squared displacements and the velocity autocorrelation functions, as well as analysing the increase of the line widths of the self-dynamic structure factors. The sodium diffusion coefficients show a remarkable good agreement between experiment and simulation utilising the polarisable potential.

  12. Degradation of methyl bromide in anaerobic sediments

    USGS Publications Warehouse

    Oremland, R.S.; Miller, L.G.; Strohmaler, F.E.

    1994-01-01

    Methyl bromide (MeBr) was anaerobically degraded in saltmarsh sediments after reaction with sulfide. The product of this nucleophilic substitution reaction was methanethiol, which underwent further chemical and bacterial reactions to form dimethyl sulfide. These two gases appeared transiently during sediment incubations because they were metabolized by methanogenic and sulfate-reducing bacteria. A second, less significant reaction of MeBr was the exchange with chloride, forming methyl chloride, which was also susceptible to attack by sulfide. Incubation of 14C-labeled methyl iodide as an analogue of MeBr resulted in the formation of 14CH4 and 14CO2 and also indicated that sulfate-reducing bacteria as well as methanogens metabolized the methylated sulfur intermediates. These results suggest that exposed sediments with abundant free sulfide, such as coastal salt-marshes, may constitute a sink for atmospheric MeBr.

  13. Indirect spectrophotometric determination of traces of bromide in water

    USGS Publications Warehouse

    Fishman, M. J.; Skougstad, M.W.

    1963-01-01

    A rapid, accurate, and sensitive indirect spectrophotometric method for the determination of bromide in natural waters is based on the catalytic effect of bromide on the oxidation of iodine to iodate by potassium permanganate in sulfuric acid solution. The method is applicable to concentrations ranging from 1 to 100 ??g. of bromide per liter, but may be modified to extend the concentration range. Most ions commonly occurring in water do not interfere. The standard deviation is 2.9 at bromide concentrations of 100 ??g. per liter and less at lower concentrations. The determination of bromide in samples containing known added amounts gave values ranging from 99 to 105% of the concentration calculated to be present.

  14. Selective oxidation of bromide in wastewater brines from hydraulic fracturing.

    PubMed

    Sun, Mei; Lowry, Gregory V; Gregory, Kelvin B

    2013-07-01

    Brines generated from oil and natural gas production, including flowback water and produced water from hydraulic fracturing of shale gas, may contain elevated concentrations of bromide (~1 g/L). Bromide is a broad concern due to the potential for forming brominated disinfection byproducts (DBPs) during drinking water treatment. Conventional treatment processes for bromide removal is costly and not specific. Selective bromide removal is technically challenging due to the presence of other ions in the brine, especially chloride as high as 30-200 g/L. This study evaluates the ability of solid graphite electrodes to selectively oxidize bromide to bromine in flowback water and produced water from a shale gas operation in Southwestern PA. The bromine can then be outgassed from the solution and recovered, as a process well understood in the bromine industry. This study revealed that bromide may be selectively and rapidly removed from oil and gas brines (~10 h(-1) m(-2) for produced water and ~60 h(-1) m(-2) for flowback water). The electrolysis occurs with a current efficiency between 60 and 90%, and the estimated energy cost is ~6 kJ/g Br. These data are similar to those for the chlor-alkali process that is commonly used for chlorine gas and sodium hydroxide production. The results demonstrate that bromide may be selectively removed from oil and gas brines to create an opportunity for environmental protection and resource recovery.

  15. Toxicity of the amphetamine metabolites 4-hydroxyamphetamine and 4-hydroxynorephedrine in human dopaminergic differentiated SH-SY5Y cells.

    PubMed

    Feio-Azevedo, R; Costa, V M; Ferreira, L M; Branco, P S; Pereira, F C; Bastos, M L; Carvalho, F; Capela, J P

    2017-03-05

    Amphetamine (AMPH) is a psychostimulant used worldwide by millions of patients in the clinical treatment of attention deficit hyperactivity disorder, narcolepsy or even obesity, and is also a drug of abuse. 4-Hydroxynorephedrine (4-OHNE) and 4-hydroxyamphetamine (4-OHAMPH) are two major metabolites known to persist in the brain longer than AMPH. The contribution of AMPH metabolites for its neurotoxicity is undetermined. We evaluated the toxicity of AMPH and its metabolites 4-OHNE and 4-OHAMPH, obtained by chemical synthesis, in human dopaminergic differentiated SH-SY5Y neurons. Cells were exposed to AMPH (concentration range 0-5mM) or 4-OHAMPH or 4-OHNE (concentration range 0-10mM) for 24 or 48h, and the viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and lactate dehydrogenase (LDH) leakage assays. Results showed that for both AMPH and the metabolites a concentration-dependent toxicity was observed. The toxic concentration 50% (TC50) for AMPH and 4-OHNE following 24h exposure was circa 3.5mM and 8mM, respectively. For 4-OHAMPH the TC50 was not reached in the tested concentration range. N-acetyl cysteine, cycloheximide, l-carnitine, and methylphenidate were able to reduce cell death induced by AMPH TC50. Acridine orange/ethidium bromide staining showed evident signs of late apoptotic cells and necrotic cells following 24h exposure to AMPH 3.50mM. The 4-OHAMPH metabolite at 8.00mM originated few late apoptotic cells, whereas 4-OHNE at 8.00mM resulted in late apoptotic cells and necrotic cells, in a scenario similar to AMPH. In conclusion, the AMPH metabolite 4-OHNE is more toxic than 4-OHAMPH, nonetheless both are less toxic than the parent compound in vitro. The most toxic metabolite 4-OHNE has longer permanence in the brain, rendering likely its contribution for AMPH neurotoxicity.

  16. In Vitro DNA-Binding, Anti-Oxidant and Anticancer Activity of Indole-2-Carboxylic Acid Dinuclear Copper(II) Complexes.

    PubMed

    Wang, Xiangcong; Yan, Maocai; Wang, Qibao; Wang, Huannan; Wang, Zhengyang; Zhao, Jiayi; Li, Jing; Zhang, Zhen

    2017-01-20

    Indole-2-carboxylic acid copper complex (ICA-Cu) was successfully prepared and characterized through elemental analysis, IR, UV-Vis, ¹H-NMR, TG analysis, and molar conductance, and its molecular formula was [Cu₂(C₉H₆O₂N)₄(H₂O)₂]·2H₂O. The binding ability of ICA-Cu to calf thymus DNA (CT-DNA) was examined by fluorescence spectrometry and the viscosity method. The results indicated that, upon the addition of increasing amounts of CT-DNA, the excitation and emission intensity of ICA-Cu decreased obviously and the excitation spectra shifted towards a long wavelength. ICA-Cu could displace ethidium bromide (EB) from the EB-DNA system, making the fluorescence intensity of the EB-DNA system decrease sharply; the quenching constant KSV value was 3.99 × 10⁴ M(-1). The emission intensity of the ICA-Cu-DNA system was nearly constant, along with the addition of Na⁺ in a series of concentrations. The fluorescence of the complex could be protected after the complex interacted with DNA. A viscosity measurement further supported the result that the ICA-Cu complex may interact with DNA in an intercalative binding mode. The antioxidant activities of ICA-Cu were evaluated by a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, a hydroxyl radical (OH) scavenging assay, and a 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay. The ICA-Cu exhibited the highest inhibitory effects on the ABTS radical (94% inhibition at 60 µM), followed by OH and DPPH radicals (the degrees of inhibition being 71% and 56%, respectively). The in vitro cytotoxicity activity of ICA-Cu against two human breast cancer cell lines, MDA-MB-231 and MCF-7, was investigated by 3-[4,5-dimethyltiazol2-yl]-2.5-diphenyl-tetrazolium bromide (MTT) assay and cellular morphological analysis. The results showed that, upon increasing the concentration of ICA-Cu, an increase was observed in growth-inhibitory activity and the inhibition percentage were greater than 90% at 20 µM in both cell

  17. Effects of Perivitelline Fluid Obtained from Horseshoe Crab on The Proliferation and Genotoxicity of Dental Pulp Stem Cells

    PubMed Central

    Musa, Marahaini; Mohd Ali, Khadijah; Kannan, Thirumulu Ponnuraj; Azlina, Ahmad; Omar, Nor Shamsuria; Chatterji, Anil; Mokhtar, Khairani Idah

    2015-01-01

    Objective Perivitelline fluid (PVF) of the horseshoe crab embryo has been reported to possess an important role during embryogenesis by promoting cell proliferation. This study aims to evaluate the effect of PVF on the proliferation, chromosome aberration (CA) and mutagenicity of the dental pulp stem cells (DPSCs). Materials and Methods This is an in vitro experimental study. PVF samples were collected from horseshoe crabs from beaches in Malaysia and the crude extract was prepared. DPSCs were treated with different concentrations of PVF crude extract in an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay (cytotoxicity test). We choose two inhibitory concentrations (IC50 and IC25) and two PVF concentrations which produced more cell viability compared to a negative control (100%) for further tests. Quantitative analysis of the proliferation activity of PVF was studied using the AlamarBlue®assay for 10 days. Population doubling times (PDTs) of the treatment groups were calculated from this assay. Genotoxicity was evaluated based on the CA and Ames tests. Statistical analysis was carried out using independent t test to calculate significant differences in the PDT and mitotic indices in the CA test between the treatment and negative control groups. Significant differences in the data were P<0.05. Results A total of four PVF concentrations retrieved from the MTT assay were 26.887 mg/ml (IC50), 14.093 mg/ml (IC25), 0.278 mg/ml (102% cell viability) and 0.019 mg/ml (102.5% cell viability). According to the AlamarBlue®assay, these PVF groups produced comparable proliferation activities compared to the negative (untreated) control. PDTs between PVF groups and the negative control were insignificantly different (P>0.05). No significant aberrations in chromosomes were observed in the PVF groups and the Ames test on the PVF showed the absence of significant positive results. Conclusion PVF from horseshoe crabs produced insignificant proliferative

  18. Treatment with Rutin - A Therapeutic Strategy for Neutrophil-Mediated Inflammatory and Autoimmune Diseases

    PubMed Central

    Nikfarjam, Bahareh Abd; Adineh, Mohtaram; Hajiali, Farid

    2017-01-01

    Objectives: Neutrophils represent the front line of human defense against infections. Immediately after stimulation, neutrophilic enzymes are activated and produce toxic mediators such as pro-inflammatory cytokines, nitric oxide (NO) and myeloperoxidase (MPO). These mediators can be toxic not only to infectious agents but also to host tissues. Because flavonoids exhibit antioxidant and anti-inflammatory effects, they are subjects of interest for pharmacological modulation of inflammation. In the present study, the effects of rutin on stimulus-induced NO and tumor necrosis factor (TNF)-α productions and MPO activity in human neutrophils were investigated. Methods: Human peripheral blood neutrophils were isolated using Ficoll-Hypaque density gradient centrifugation coupled with dextran T500 sedimentation. The cell preparations containing > 98% granulocytes were determined by morphological examination through Giemsa staining. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without rutin (25 μM) for 45 minutes, and stimulated with phorbol 12-myristate 13-acetate (PMA). Then, the TNF-α, NO and MPO productions were analyzed using enzyme-linked immunosorbent assay (ELISA), Griess Reagent, and MPO assay kits, respectively. Also, the viability of human neutrophils was assessed using tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and neutrophils were treated with various concentrations of rutin (1 - 100 μM), after which MTT was appended and incubated at 37ºC for 4 hour. Results: Rutin at concentrations up to 100 μM did not affect neutrophil viability during the 4-hour incubation period. Rutin significantly decreased the NO and TNF-α productions in human peripheral blood neutrophils compared to PMA-control cells (P < 0.001). Also, MPO activity was significantly reduced by rutin (P < 0.001). Conclusion: In this in vitro study, rutin had an anti-inflammatory effect due to

  19. Production of transgenic canine embryos using interspecies somatic cell nuclear transfer.

    PubMed

    Hong, So Gun; Oh, Hyun Ju; Park, Jung Eun; Kim, Min Jung; Kim, Geon A; Koo, Ok Jae; Jang, Goo; Lee, Byeong Chun

    2012-02-01

    Somatic cell nuclear transfer (SCNT) has emerged as an important tool for producing transgenic animals and deriving transgenic embryonic stem cells. The process of SCNT involves fusion of in vitro matured oocytes with somatic cells to make embryos that are transgenic when the nuclear donor somatic cells carry 'foreign' DNA and are clones when all the donor cells are genetically identical. However, in canines, it is difficult to obtain enough mature oocytes for successful SCNT due to the very low efficiency of in vitro oocyte maturation in this species that hinders canine transgenic cloning. One solution is to use oocytes from a different species or even a different genus, such as bovine oocytes, that can be matured easily in vitro. Accordingly, the aim of this study was: (1) to establish a canine fetal fibroblast line transfected with the green fluorescent protein (GFP) gene; and (2) to investigate in vitro embryonic development of canine cloned embryos derived from transgenic and non-transgenic cell lines using bovine in vitro matured oocytes. Canine fetal fibroblasts were transfected with constructs containing the GFP and puromycin resistance genes using FuGENE 6®. Viability levels of these cells were determined by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. Interspecies SCNT (iSCNT) embryos from normal or transfected cells were produced and cultured in vitro. The MTT measurement of GFP-transfected fetal fibroblasts (mean OD = 0.25) was not significantly different from non-transfected fetal fibroblasts (mean OD = 0.35). There was no difference between transgenic iSCNT versus non-transgenic iSCNT embryos in terms of fusion rates (73.1% and 75.7%, respectively), cleavage rates (69.7% vs. 73.8%) and development to the 8-16-cell stage (40.1% vs. 42.7%). Embryos derived from the transfected cells completely expressed GFP at the 2-cell, 4-cell, and 8-16-cell stages without mosaicism. In summary, our results demonstrated that

  20. In vitro cytotoxicity assessment of two heavy metal salts in a fish cell line (RF).

    PubMed

    Goswami, Mukunda; Yadav, Kamalendra; Dubey, Akhilesh; Sharma, Bhagwati S; Konwar, Rituraj; Kumar, Ravindra; Nagpure, Naresh S; Lakra, Wazir S

    2014-01-01

    Labeo rohita (rohu) is one of most important of Indian major carps, which is highly ranked among the important freshwater aquaculture species in the world. Heavy metals are major risk factors for aquatic health, which are biomagnified through the food chain. The present study was carried out to investigate the effect of different divalent salts of two heavy metals, such as zinc (ZnCl(2), ZnSO(4) and ZnNO(3)) and cadmium (CdCl(2) and CdSO(4)), in an established fish cell line, RF developed from fin tissue of L. rohita. The RF cell line was used for assessment of heavy metal cytotoxicity through various endpoint assays, including maximum tolerated dose (MTD) determination, 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay, neutral red (NR) uptake assay, and Coomassie brilliant blue (CBB) assay. Results revealed that these heavy metal salts were cytotoxic to the RF cell line at varied concentrations. MTD values were found to be 1.563, 3.125, 6.25, 12.5 and 25 mg/L for CdCl(2), CdSO(4), ZnCl(2), Zn(NO(3))(2) and ZnSO(4), respectively. The half-maximal inhibitory concentration values calculated by MTT, NR and CB assay were 53.83 ± 7.02, 58.03 ± 9.12 and 79.20 ± 15.27 for ZnSO(4), 26.44 ± 7.01, 36.60 ± 7.82 and 155.6 ± 14.75 for Zn(NO(3))(2), 20.26 ± 17.95, 16.94 ± 7.05 and 87.54 ± 7.58 for ZnCl(2), 5.166 ± 0.57, 15 ± 1 and 41.80 ± 8.38 for CdSO(4) and 4.966 ± 0.56, 9.56 ± 1.73 and 20.93 ± 4.47 for CdCl(2). This study establishes the RF cell line as an in vitro tool for assessment and monitoring of heavy metal concentration in the aquatic environment.

  1. Antioxidant and Nephroprotective Activities of the Extract and Fractions of Homonoia riparia Lour

    PubMed Central

    Xavier, Seena kanniparambil; Haneefa, Shoja Muhammed; Anand, Devkar Raviraj; polo, Picheswara Rao; Maheshwari, Rajalekshmi; Shreedhara, Chandrashekara Shastry; Setty, Manganahalli Manjunath

    2017-01-01

    -(4,5-dimethylthiazolyl-2-yl)-2,5- diphenyl tetrazolium bromide, GAE: Gallic acid equivalents, QE: Quercetin equivalents, HEK: Human Embryonic Kidney, HRM: Methanol extract of H. riparia, HRPE: Petroleum ether fraction of H. riparia, HREA: ethyl acetate fraction of H. riparia, HRBU: Butanol fraction of H. riparia, HRAQ: Aqueous fraction of H. riparia, DMEM: Dulbecco's minimum essential medium, FBS: Foetal bovine serum, DMSO: Dimethyl sulfoxide, ANOVA: One way analysis of variance. PMID:28216879

  2. Chloride, bromide and iodide scintillators with europium

    DOEpatents

    Zhuravleva, Mariya; Yang, Kan

    2016-09-27

    A halide scintillator material is disclosed where the halide may comprise chloride, bromide or iodide. The material is single-crystalline and has a composition of the general formula ABX.sub.3 where A is an alkali, B is an alkali earth and X is a halide which general composition was investigated. In particular, crystals of the formula ACa.sub.1-yEu.sub.yI.sub.3 where A=K, Rb and Cs were formed as well as crystals of the formula CsA.sub.1-yEu.sub.yX.sub.3 (where A=Ca, Sr, Ba, or a combination thereof and X=Cl, Br or I or a combination thereof) with divalent Europium doping where 0.ltoreq.y.ltoreq.1, and more particularly Eu doping has been studied at one to ten mol %. The disclosed scintillator materials are suitable for making scintillation detectors used in applications such as medical imaging and homeland security.

  3. Methyl Bromide Commodity Fumigation Buffer Zone Lookup Tables

    EPA Pesticide Factsheets

    Product labels for methyl bromide used in commodity and structural fumigation include requirements for buffer zones around treated areas. The information on this page will allow you to find the appropriate buffer zone for your planned application.

  4. 7 CFR 305.6 - Methyl bromide fumigation treatment schedules.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ..., fumigation with methyl bromide for sapote fruit fly. Regulated citrus fruits originating inside an area quarantined for sapote fruit fly that are to be moved outside the quarantined area may be treated with...

  5. Computational and vibrational spectroscopic studies of ipratropium bromide.

    PubMed

    Ali, H R H; Edwards, H G M; Kendrick, J; Scowen, I J

    2009-02-01

    In this study, ipratropium bromide is investigated using vibrational spectroscopy and quantum chemical calculations. The structure of ipratropium bromide was optimised using density functional theory calculations and the geometry optimisation has been carried out on two conformations with and without intramolecular hydrogen bonding. Infrared and Raman spectra were calculated from the optimised structures. Many modes in the calculated spectra could be matched with the experimental spectra and a description of the modes is given. By analysis of the theoretical vibrational modes, it is shown that ipratropium bromide specimens are likely to be a mixture of the two conformations with and without intramolecular hydrogen bonding. In addition, several spectral features and band intensities in the CH and OH stretching regions are explained. Quantum mechanical calculations allowed improved understanding of ipratropium bromide and its vibrational spectra.

  6. Pd-Catalyzed Nucleophilic Fluorination of Aryl Bromides

    PubMed Central

    2015-01-01

    On the basis of mechanism-driven reaction design, a Pd-catalyzed nucleophilic fluorination of aryl bromides and iodides has been developed. The method exhibits a broad substrate scope, especially with respect to nitrogen-containing heteroaryl bromides, and proceeds with minimal formation of the corresponding reduction products. A facilitated ligand modification process was shown to be critical to the success of the reaction. PMID:24559304

  7. Synthesis, self-aggregation and biological properties of alkylphosphocholine and alkylphosphohomocholine derivatives of cetyltrimethylammonium bromide, cetylpyridinium bromide, benzalkonium bromide (C16) and benzethonium chloride.

    PubMed

    Lukáč, Miloš; Mrva, Martin; Garajová, Mária; Mojžišová, Gabriela; Varinská, Lenka; Mojžiš, Ján; Sabol, Marián; Kubincová, Janka; Haragová, Hana; Ondriska, František; Devínsky, Ferdinand

    2013-08-01

    A series of alkylphosphocholine and alkylphosphohomocholine derivatives of cetyltrimethylammonium bromide, cetylpyridinium bromide, benzalkonium bromide (C16) and benzethonium chloride have been synthesized. Their physicochemical properties were also investigated. The critical micelle concentration (cmc), the surface tension value at the cmc (γcmc), and the surface area at the surface saturation per head group (Acmc) were determined by means of surface tension measurements. The prepared compounds exhibit significant cytotoxic, antifungal and antiprotozoal activities. Alkylphosphocholines and alkylphosphohomocholines possess higher antifungal activity against Candida albicans in comparison with quaternary ammonium compounds in general. However, quaternary ammonium compounds exhibit significantly higher activity against human tumor cells and pathogenic free-living amoebae Acanthamoeba lugdunensis and Acanthamoeba quina compared to alkylphosphocholines. The relationship between structure, physicochemical properties and biological activity of the tested compounds is discussed.

  8. Noncolorimetric measurement of cell activity in three-dimensional histoculture using the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide: the pixel image analysis of formazan crystals.

    PubMed

    Colangelo, D; Guo, H Y; Connors, K M; Silvestro, L; Hoffman, R M

    1992-08-15

    We describe a novel system for measuring the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction in three-dimensional histoculture which is no longer dependent on colorimetric determination of extracted formazan, but rather is based on a pixel image analysis of formazan crystals, and which allows intratumor heterogeneity to be taken into account. The MTT test is based on the enzymatic reduction of the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-dipheniltetrazolium bromide to formazan crystals by living, metabolically active cells, but not in dead cells. The reaction was carried out in situ in six-well plates on gel-supported histocultured human tumors. After a 24-h incubation with different drugs the tumors were incubated with a solution of MTT. Frozen sections of the tumor pieces were made and the slides were then stained with a propidium iodide solution, whose fluorescence is proportional to the number of cells present. We demonstrate here that the formazan crystals, formed by MTT reduction, reflect polarized light and that this can be quantified by using an image analysis system based on bright-pixel quantitation directly on a frozen section of the original tissue. Combined with the use of the fluorescent dye propidium iodide, also measured by pixel analysis, we can express a ratio between the total amount of MTT reduction and the total number of cells present in the specimen that expresses the effect of drugs on the histocultured tumors. Since histology is well maintained in histoculture it is possible to take into account the heterogeneity present in the tumor with regard to drug response.(ABSTRACT TRUNCATED AT 250 WORDS)

  9. Spectral characteristics of the bentonite loaded with benzyldimethyloctadecylammonium chloride, hexadecyltrimethylammonium bromide and dimethyldioctadecylammonium bromide

    NASA Astrophysics Data System (ADS)

    Majdan, Marek; Maryuk, Oksana; Gładysz-Płaska, Agnieszka; Pikus, Stanisław; Kwiatkowski, Ryszard

    2008-02-01

    The spectral characterization, including the FTIR, DRIFT (diffusive reflectance), SWAXS (small and wide angle X-ray scattering) spectra comparison of the sodium bentonite modified by BDMODA-Cl (benzyldimethyloctadecylammonium chloride), HDTMA-Br (hexadecyltrimethylammonium bromide), DDA-Br (dimethyldioctadecylammonium bromide) is presented in the paper. The FTIR spectra show the shift of C-H stretching vibrations: νsym(CH2), νasym(CH2) of surfactants methylene chains toward lower frequencies (from 2855 to 2851 cm -1 for νsym(CH2) and from 2927 to 2918 cm -1 for νansym(CH2) with the surfactant concentration in bentonite phase. The bending vibrations δH-O-H in water molecules change their positions in the direction of higher frequencies (from 1634 to 1647 cm -1) with the surfactant concentration for bentonite-BDMODA and bentonite-DDA contrary to bentonite-HDTMA, where the constant position δH-O-H is explained as the consequence of the lower concentration of the hydrogen bonded water in bentonite-HDTMA phase when compared with the remaining forms of bentonite. The DRIFT spectra reveal dramatic shift of the νSi-O stretching vibration toward higher frequencies upon intercalation of the sodium bentonite with the surfactant cations. The SWAXS spectra and SEM images of the bentonite are the evidence of somewhat different sorption mechanism of DDA-Br when compared with the BDMODA-Cl and HDTMA-Br, including remarkable external surface sorption contribution in the overall sorption.

  10. 49 CFR 173.193 - Bromoacetone, methyl bromide, chloropicrin and methyl bromide or methyl chloride mixtures, etc.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... pounds) water capacity (nominal). This capacity does not apply to shipments of methyl bromide. (c) Methyl... metal cans containing not over one pound each, or inside metal cans with a minimum wall thickness of...

  11. Alkene- and alkyne- substituted methylimidazolium bromides: structural effects and Physical properties (Postprint)

    DTIC Science & Technology

    2007-08-01

    1, 3 - butene - 2 , propargyl- 3 , 2 -butyne- 4, and 2 -pent- yne- 5 side chains and their structural and physical proper- ties have been investigated...pargyl bromide, 2 -butyne bromide, 3 - butene bromide and 2 -pen- tyne bromide), methanol (99.93 % A.C.S. HPLC grade) and diethyl ether (anhydrous, 99...zolium halides (X Cl, Br, I), and 1-( 2 -butyne)- 3 - methyl - imidazolium bromide have been mentioned earlier [20, 21, 2328, 38, 39]. However, detailed

  12. Cytotoxic potential of few Indian fruit peels through 3-(4,5-dimethylthiazol-yl)-2,5-diphenyltetrazolium bromide assay on HepG2 cells

    PubMed Central

    Garg, Munish; Lata, Kusum; Satija, Saurabh

    2016-01-01

    Objective: To investigate in vitro anticancer activity of a few Indian fruit peels through 3-(4,5-dimethylthiazol-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against HepG2 cells. Materials and Methods: Hydroalcoholic extracts were prepared of five fruit peels, i.e., banana, lemon, guava, orange, and papaya by maceration and thereafter subjected for MTT assay to evaluate anticancer potential on HepG2 cells. Plant extract showed best activity was further fractionated with petroleum ether, chloroform, and ethyl acetate successively and screened again. Phytochemical analysis was then carried out to find out responsible components for the observed activity. Results: Out of the 40 samples from five fruit peel extracts with rich folklore usage, papaya extract showed maximum activity with least inhibitory concentration50 (IC50) value of 18.5 μg/ml. Further analysis after fractionation of the papaya peel extract, aqueous fraction showed the maximum inhibitory activity with least IC50 value of 17.3 μg/ml. Phytochemical analysis of the aqueous fraction of papaya peel extract revealed the presence of flavonoids and glycosides. Total flavonoid content found to be 72.25 mg/g. Conclusion: Papaya fruit extract demonstrated the best activity against MTT assay which may be due to the presence of flavonoids. PMID:26997725

  13. Pitfalls of the MTT assay: Direct and off-target effects of inhibitors can result in over/underestimation of cell viability.

    PubMed

    Stepanenko, A A; Dmitrenko, V V

    2015-12-15

    The MTT assay (to a less degree MTS, XTT or WST) is a widely exploited approach for measuring cell viability/drug cytotoxicity. MTT reduction occurs throughout a cell and can be significantly affected by a number of factors, including metabolic and energy perturbations, changes in the activity of oxidoreductases, endo-/exocytosis and intracellular trafficking. Over/underestimation of cell viability by the MTT assay may be due to both adaptive metabolic and mitochondrial reprogramming of cells subjected to drug treatment-mediated stress and inhibitor off-target effects. Previously, imatinib, rottlerin, ursolic acid, verapamil, resveratrol, genistein nanoparticles and some polypeptides were shown to interfere with MTT reduction rate resulting in inconsistent results between the MTT assay and alternative assays. Here, to test the under/overestimation of viability by the MTT assay, we compared results derived from the MTT assay with the trypan blue exclusion assay after treatment of glioblastoma U251, T98G and C6 cells with three widely used inhibitors with the known direct and side effects on energy and metabolic homeostasis - temozolomide (TMZ), a DNA-methylating agent, temsirolimus (TEM), an inhibitor of mTOR kinase, and U0126, an inhibitor of MEK1/2 kinases. Inhibitors were applied shortly as in IC50 evaluating studies or long as in studies focusing on drug resistance acquisition. We showed that over/underestimation of cell viability by the MTT assay and its significance depends on a cell line, a time point of viability measurement and other experimental parameters. Furthermore, we provided a comprehensive survey of factors that should be accounted in the MTT assay. To avoid result misinterpretation, supplementation of the tetrazolium salt-based assays with other non-metabolic assays is recommended.

  14. Zinc Bromide Combustion: Implications for the Consolidated Incinerator Facility

    SciTech Connect

    Oji, L.N.

    1998-12-16

    In the nuclear industry, zinc bromide (ZnBr2) is used for radiation shielding. At Savannah River Site (SRS) zinc bromide solution, in appropriate configurations and housings, was used mainly for shielding in viewing windows in nuclear reactor and separation areas. Waste stream feeds that will be incinerated at the CIF will occasionally include zinc bromide solution/gel matrices.The CIF air pollution systems control uses a water-quench and steam atomizer scrubber that collects salts, ash and trace metals in the liquid phase. Water is re-circulated in the quench unit until a predetermined amount of suspended solids or dissolved salts are present. After reaching the threshold limit, "dirty liquid", also called "blowdown", is pumped to a storage tank in preparation for treatment and disposal. The air pollution control system is coupled to a HEPA pre-filter/filter unit, which removes particulate matter from the flue gas stream (1).The objective of this report is to review existing literature data on the stability of zinc bromide (ZnBr2) at CIF operating temperatures (>870 degrees C (1600 degrees F) and determine what the combustion products are in the presence of excess air. The partitioning of the combustion products among the quencher/scrubber solution, bottom ash and stack will also be evaluated. In this report, side reactions between zinc bromide and its combustion products with fuel oil were not taken into consideration.

  15. Methyl bromide: effective pest management tool and environmental threat.

    PubMed

    Thomas, W B

    1996-12-01

    Methyl bromide is used extensively on a global basis as a pesticide against nematodes, weeds, insects, fungi, bacteria, and rodents. As a soil fumigant, it is used in significant quantities in the production of strawberry and tomato, as well as other agriculture commodities. Grain, fresh fruit, forestry products, and other materials are fumigated with methyl bromide to control pest infestations during transport and storage. Structures also are treated with this chemical to control wood-destroying insects and rodents. However, methyl bromide has been identified as a significant ozone-depleting substance, resulting in regulatory actions being taken by the U.S. Environmental Protection Agency and the United Nations Environment Program (Montreal Protocol). The science linking methyl bromide to ozone depletion is strong and was reinforced by the 1994 UNEP Montreal Protocol Science Assessment on Ozone Depletion, which states, "Methyl bromide continues to be viewed as a significant ozone-depleting compound." Identifying efficacious and viable alternatives in the near term is critical.

  16. Cross-coupling of aromatic bromides with allylic silanolate salts.

    PubMed

    Denmark, Scott E; Werner, Nathan S

    2008-12-03

    The sodium salts of allyldimethylsilanol and 2-butenyldimethylsilanol undergo palladium-catalyzed cross-coupling with a wide variety of aryl bromides to afford allylated and crotylated arenes. The coupling of both silanolates required extensive optimization to deliver the expected products in high yields. The reaction of the allyldimethylsilanolate takes place at 85 degrees C in 1,2-dimethoxyethane with allylpalladium chloride dimer (2.5 mol %) to afford 73-95% yields of the allylation products. Both electron-rich and sterically hindered bromides reacted smoothly, whereas electron-poor bromides cross-coupled in poor yield because of a secondary isomerization to the 1-propenyl isomer (and subsequent polymerization). The 2-butenyldimethylsilanolate (E/Z, 80:20) required additional optimization to maximize the formation of the branched (gamma-substitution) product. A remarkable influence of added alkenes (dibenzylideneacetone and norbornadiene) led to good selectivities for electron-rich and electron-poor bromides in 40-83% yields. However, bromides containing coordinating groups (particularly in the ortho position) gave lower, and in one case even reversed, selectivity. Configurationally homogeneous (E)-silanolates gave slightly higher gamma-selectivity than the pure (Z)-silanolates. A unified mechanistic picture involving initial gamma-transmetalation followed by direct reductive elimination or sigma-pi isomerization can rationalize all of the observed trends.

  17. Methyl Bromide: Effective Pest Management Tool and Environmental Threat

    PubMed Central

    Thomas, W. B.

    1996-01-01

    Methyl bromide is used extensively on a global basis as a pesticide against nematodes, weeds, insects, fungi, bacteria, and rodents. As a soil fumigant, it is used in significant quantities in the production of strawberry and tomato, as well as other agriculture commodities. Grain, fresh fruit, forestry products, and other materials are fumigated with methyl bromide to control pest infestations during transport and storage. Structures also are treated with this chemical to control wood-destroying insects and rodents. However, methyl bromide has been identified as a significant ozone-depleting substance, resulting in regulatory actions being taken by the U.S. Environmental Protection Agency and the United Nations Environment Program (Montreal Protocol). The science linking methyl bromide to ozone depletion is strong and was reinforced by the 1994 UNEP Montreal Protocol Science Assessment on Ozone Depletion, which states, "Methyl bromide continues to be viewed as a significant ozone-depleting compound." Identifying efficacious and viable alternatives in the near term is critical. PMID:19277178

  18. Cetyltrimethyl ammonium bromide assisted hydrothermal growth of hematite hollow cubes

    SciTech Connect

    Wang, Wei-Wei; Yao, Jia-Liang

    2010-11-15

    Hematite hollow cubes have been prepared by forced hydrolysis of ferric chloride solutions under hydrothermal conditions. The effects of reaction time, reaction temperature and cetyltrimethyl ammonium bromide on the transformation process from akageneite to hematite were investigated in detail. The products were characterized by X-ray powder diffraction, scanning electron microscopy and transmission electron microscopy. It is found that cetyltrimethyl ammonium bromide was a critical factor influencing the phase transformation process of akageneite and the final morphology of the as-prepared products. With cetyltrimethyl ammonium bromide, hematite hollow cubes and porous spheres were obtained. Otherwise only dense cubes were observed even prolonging reaction time or increasing reaction temperature. The mechanism was proposed.

  19. [Tiotropium bromide for treating chronic obstructive pulmonary disease].

    PubMed

    Uteshev, D B; Buniatian, N D; Kovaleva, V L

    2010-11-01

    Five different types of muscarine-sensitive receptors were identified until now. In routine practice, the nonselective antagonist of cholinoreceptors are replaced by ipratropium bromide that is selectively blocking M1, M2, and M3 subtypes with the same affinity to each of them. However, the blockage of M2 subtype leads to bronchoconstriction and is accompanied by inhibition of M3 receptors in bronchial smooth muscles. The new drug tiotropium bromide selectively inhibits only the M1 and M3 types of receptors and does not affect the M2 subtype. This drug is administered only once a day, which is very important in clinical practice. Thus tiotropium bromide is the drug of choice for basic therapy of COPD.

  20. Sodium cromoglycate and ipratropium bromide in exercise-induced asthma.

    PubMed Central

    Thomson, N C; Patel, K R; Kerr, J W

    1978-01-01

    In thirteen patients with extrinsic asthma the effects of placebo, sodium cromoglycate, ipratropium bromide, and ipratropium bromide plus sodium cromoglycate were studied in a random double-blind fashion to assess their inhibitory action in exercise-induced asthma (EIA). Exercise testing consisted of steady state running on an inclined treadmill for up to eight minutes. In eight of the 13 patients studied the baseline ratio of expiratory flow at 50% vital capacity (VC) breathing helium-oxygen (V50He) to V50air was over 1.20 and they were called responders; the remaining five patients were called non-responders. There was a significantly lower baseline maximum mid-expiratory flow rate (MMEF) in non-responders (P less than 0.02) as compared to responders but no difference in forced expiratory volume in one second (FEV1) or forced vital capacity (FVC). Sodium cromoglycate (P less than 0.02), ipratropium bromide (P less than 0.01), and ipratropium bromide plus spdium cromoglycate (P less than 0.01) all significantly inhibited the percentage fall in FEV1 after exercise in the responders. Ipratropium bromide had no preventive action on non-responders, unlike sodium cromoglycate (P less than 0.05) and ipratropium bromide plus sodium cromoglycate (P less than 0.02). It is postulated that mediator release is an important factor in development of EIA in most extrinsic asthmatics, whereas cholinergic mechanisms are relevant only in those patients in whom the main site of airflow obstruction is in the large central airways. PMID:154747

  1. Photochemistry of alkyl bromides trapped in water ice films

    NASA Astrophysics Data System (ADS)

    Schrems, O.; Okaikwei, B.; Bluszcz, Th.

    2012-04-01

    Photochemical reactions of atmospheric trace gases taking place at the surface of atmospheric ice particles and in bulk ice are important in stratospheric and tropospheric chemistry but also in polar and alpine snowpack chemistry. Consequently, the understanding of the uptake und incorporation of atmospheric trace gases in water ice as well as their interactions with water molecules is very important for the understanding of processes which occur in ice particles and at the air/ice interface. Reactive atmospheric trace gases trapped in ice are subject of photochemical reactions when irradiated with solar UV radiation. Among such compounds bromine species are highly interesting due to their potential of depleting ozone both in the stratosphere and troposphere. Organic bromine gases can carry bromine to the stratosphere. Methyl bromide (CH3Br) is the largest bromine carrier to the stratosphere. It has both natural and anthropogenic sources. In this contribution we will present the results of our laboratory studies of alkyl bromides (methyl, bromide (CH3Br), dimethyl bromide (CH2Br2), n-propyl bromide (C3H7Br), 1,2-dibromoethane C2H4Br2)), trapped in water ice. We have simulated the UV photochemistry of these brominated alkanes isolated in ice films kept at 16 K and for comparison in solid argon matrices. The photoproducts formed in the ice have been identified by means of FTIR spectroscopy. Reflection absorption infrared spectroscopy (RAIRS) is especially useful to study nascent ice surfaces, kinetics of adsorption/decomposition, and heterogeneous catalysis. Among the observed photoproducts we could identify carbon monoxide and carbon dioxide for each alkyl bromide studied. The photoproduct HBr is dissociated in the bulk ice. Based on the experimental observations possible reaction mechanisms will be discussed.

  2. Antimalarial Drug Artemether Inhibits Neuroinflammation in BV2 Microglia Through Nrf2-Dependent Mechanisms.

    PubMed

    Okorji, Uchechukwu P; Velagapudi, Ravikanth; El-Bakoush, Abdelmeneim; Fiebich, Bernd L; Olajide, Olumayokun A

    2016-11-01

    Artemether, a lipid-soluble derivative of artemisinin has been reported to possess anti-inflammatory properties. In this study, we have investigated the molecular mechanisms involved in the inhibition of neuroinflammation by the drug. The effects of artemether on neuroinflammation-mediated HT22 neuronal toxicity were also investigated in a BV2 microglia/HT22 neuron co-culture. To investigate effects on neuroinflammation, we used LPS-stimulated BV2 microglia treated with artemether (5-40 μM) for 24 h. ELISAs and western blotting were used to detect pro-inflammatory cytokines, nitric oxide, prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and microsomal prostaglandin E synthase-1 (mPGES-1). Beta-site amyloid precursor protein cleaving enzyme 1 (BACE-1) activity and Aβ levels were measured with ELISA kits. Protein levels of targets in nuclear factor kappa B (NF-κB) and p38 mitogen-activated protein kinase (MAPK) signalling, as well as heme oxygenase-1 (HO-1), NQO1 and nuclear factor-erythroid 2-related factor 2 (Nrf2) were also measured with western blot. NF-κB binding to the DNA was investigated using electrophoretic mobility shift assays (EMSA). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), DNA fragmentation and reactive oxygen species (ROS) assays in BV2-HT22 neuronal co-culture were used to evaluate the effects of artemether on neuroinflammation-induced neuronal death. The role of Nrf2 in the anti-inflammatory activity of artemether was investigated in BV2 cells transfected with Nrf2 siRNA. Artemether significantly suppressed pro-inflammatory mediators (NO/iNOS, PGE2/COX-2/mPGES-1, tumour necrosis factor-alpha (TNFα) and interleukin (IL)-6); Aβ and BACE-1 in BV2 cells following LPS stimulation. These effects of artemether were shown to be mediated through inhibition of NF-κB and p38 MAPK signalling. Artemether produced increased levels of HO-1, NQO1 and GSH in BV2 microglia. The drug activated

  3. Toxic and inflammatory effects of filters frequently used for the collection of airborne particulate matter

    NASA Astrophysics Data System (ADS)

    Dick, C. A. J.; Stone, V.; Brown, D. M.; Watt, M.; Cherrie, J. W.; Howarth, S.; Seaton, A.; Donaldson, K.

    Epidemiological studies indicate that exacerbations of airway diseases and cardio-respiratory mortality have been associated with high levels of particulate air pollution. In the search for explanatory mechanisms, there is a need to collect particulate matter (such as PM 10/PM 2.5) onto filters for biological analysis. The removal of airborne particulate matter from a filter into solution involves agitation of the samples to release the particles and this is done by such methods as high-speed vortexing. Since it is possible that components of filters could contaminate the preparation and interfere with biological investigations it is necessary to find an inert filter suitable for both particulate collection and biological analysis. Extracts of the following six filters were assessed for toxicological activity both in vivo and in vitro:-Millipore AP20, Whatman GF/A, Whatman WCN, Gelman TF1000, Gelman DM800 and polycarbonate (PC). Treatment with the extract of either Whatman GF/A, Gelman DM800 or Millipore AP20 filters for 24 h significantly ( p<0.001) decreased the metabolic competence of A549 cells, as measured by MTT (3-(4,5-dimetholthiazol-2,5-diphenyl tetrazolium bromide) reduction. In addition instillation of filter extracts into rat lungs was carried out to investigate their ability to cause inflammation. The cellular components of the bronchoalveolar lavage fluid (BALF) were analysed for the number of neutrophils, as markers of inflammation. After 24 h the filter extract of the Millipore AP20, polycarbonate (PC), Whatman WCN and Whatman GF/A induced a significant ( P<0.001) influx of neutrophils into the rat lung. Finally the ability of each filter to absorb phosphate buffer solution (PBS) was assessed. The Whatman GF/A increased in weight 13 fold and the Millipore AP20 8 fold. In comparison the polycarbonate and the Gelman TF1000 were hydrophobic. These results suggest that of the six filters tested, the Teflon Gelman TF1000 filter was the most suitable for

  4. Silencing Bag-1 gene via magnetic gold nanoparticle-delivered siRNA plasmid for colorectal cancer therapy in vivo and in vitro.

    PubMed

    Huang, Wenbai; Liu, Zhan'ao; Zhou, Guanzhou; Ling, Jianmin; Tian, Ailing; Sun, Nianfeng

    2016-08-01

    Apoptosis disorder is generally regarded as an important mechanism of carcinogenesis. Inducement of tumor cell apoptosis can be an effectual way to treat cancer. Bcl-2-associated athanogene 1 (Bag-1) is a positive regulator of Bcl-2 which is an anti-apoptotic gene. Bag-1 is highly expressed in colorectal cancer, which plays a critical role in promoting metastasis, poor prognosis, especially in anti-apoptotic function, and is perhaps a valuable gene target for colorectal cancer therapy. Recently, we applied a novel non-viral gene carrier, magnetic gold nanoparticle, and mediated plasmid pGPH1/GFP/Neo-Bag-1-homo-825 silencing Bag-1 gene for treating colorectal cancer in vivo and in vitro. By mediating with magnetic gold nanoparticle, siRNA plasmid was successfully transfected into cell. In 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, magnetic gold nanoparticle had no significant cytotoxicity and by which delivered RNA plasmid inhibited cell viability significantly (P < 0.05). Downregulation of Bag-1 promoted cell apoptosis (∼47.0 %) in vitro and significantly decreased tumor growth when the cells were injected into nude mice. Based on the studies in vivo, the relative expression of Bag-1 was 0.165 ± 0.072 at mRNA level and ∼60 % at protein level. In further study, C-myc and β-catenin, mainly molecules of Wnt/β-catenin pathway, were decreased notably when Bag-1 were silenced in nanoparticle plasmid complex-transfected Balb c/nude tumor xenograft. In conclusion, Bag-1 is confirmed an anti-apoptosis gene that functioned in colorectal cancer, and the mechanism of Bag-1 gene causing colorectal cancer may be related to Wnt/β-catenin signaling pathway abnormality and suggested that magnetic gold nanoparticle-delivered siRNA plasmid silencing Bag-1 is an effective gene therapy method for colorectal cancer.

  5. Anti-malarial activity of indole alkaloids isolated from Aspidosperma olivaceum

    PubMed Central

    2014-01-01

    Background Several species of Aspidosperma (Apocynaceae) are used as treatments for human diseases in the tropics. Aspidosperma olivaceum, which is used to treat fevers in some regions of Brazil, contains the monoterpenoid indole alkaloids (MIAs) aspidoscarpine, uleine, apparicine, and N-methyl-tetrahydrolivacine. Using bio-guided fractionation and cytotoxicity testing in a human hepatoma cell line, several plant fractions and compounds purified from the bark and leaves of the plant were characterized for specific therapeutic activity (and selectivity index, SI) in vitro against the blood forms of Plasmodium falciparum. Methods The activity of A. olivaceum extracts, fractions, and isolated compounds was evaluated against chloroquine (CQ)-resistant P. falciparum blood parasites by in vitro testing with radiolabelled [3H]-hypoxanthine and a monoclonal anti-histidine-rich protein (HRPII) antibody. The cytotoxicity of these fractions and compounds was evaluated in a human hepatoma cell line using a 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, and the SI was calculated as the ratio between the toxicity and activity. Two leaf fractions were tested in mice with Plasmodium berghei. Results All six fractions from the bark and leaf extracts were active in vitro at low doses (IC50 < 5.0 μg/mL) using the anti-HRPII test, and only two (the neutral and basic bark fractions) were toxic to a human cell line (HepG2). The most promising fractions were the crude leaf extract and its basic residue, which had SIs above 50. Among the four pure compounds evaluated, aspidoscarpine in the bark and leaf extracts showed the highest SI at 56; this compound, therefore, represents a possible anti-malarial drug that requires further study. The acidic leaf fraction administered by gavage to mice with blood-induced malaria was also active. Conclusion Using a bio-monitoring approach, it was possible to attribute the anti-P. falciparum activity of A. olivaceum to

  6. Inducing the cell cycle arrest and apoptosis of oral KB carcinoma cells by hydroxychavicol: roles of glutathione and reactive oxygen species

    PubMed Central

    Chang, M C; Uang, B J; Wu, H L; Lee, J J; Hahn, L J; Jeng, J H

    2002-01-01

    Hydroxychavicol (HC; 10 – 50 μM), a betel leaf component, was found to suppress the 2% H2O2-induced lucigenin chemiluminescence for 53 – 75%. HC (0.02 – 2 μM) was also able to trap superoxide radicals generated by a xanthine/xanthine oxidase system with 38 – 94% of inhibition. Hydroxyl radicals-induced PUC18 plasmid DNA breaks was prevented by HC (1.6 – 16 μM).A 24-h exposure of KB cells to HC (0.5, 1 mM) resulted in 54 – 74% cell death as analysed by a 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. HC (10, 50 μM) further suppressed the growth of KB cells (15 and 76%, respectively). Long-term colony formation of KB cells was inhibited by 51% with 10 μM HC.Pretreatment of KB cells with 100 μM HC inhibited the attachment of KB cells to type I collagen and fibronectin by 59 and 29%, respectively.Exposure of KB cells to 0.1 mM HC for 24 h resulted in cell cycle arrest at late S and G2/M phase. Increasing the HC concentration to 0.25 and 0.5 mM led to apoptosis as revealed by detection of sub-G0/G1 peaks with a concomitant decrease in the number of cells residing in late S and G2/M phase.Inducing the apoptosis of KB cells by HC was accompanied by marked depletion in reduced form of GSH (>0.2 mM) and the increasing of reactive oxygen species production (>0.1 mM) as analysed by CMF- and DCF-single cell fluorescence flow cytometry.These results indicate that HC exerts antioxidant property at low concentration. HC also inhibits the growth, adhesion and cell cycle progression of KB cells, whereas its induction of KB cell apoptosis (HC>0.1 mM) was accompanied by cellular redox changes. PMID:11834609

  7. L-Ascorbate attenuates methamphetamine neurotoxicity through enhancing the induction of endogenous heme oxygenase-1

    SciTech Connect

    Huang, Ya-Ni; Wang, Jiz-Yuh; Lee, Ching-Tien; Lin, Chih-Hung; Lai, Chien-Cheng; Wang, Jia-Yi

    2012-12-01

    Methamphetamine (METH) is a drug of abuse which causes neurotoxicity and increased risk of developing neurodegenerative diseases. We previously found that METH induces heme oxygenase (HO)-1 expression in neurons and glial cells, and this offers partial protection against METH toxicity. In this study, we investigated the effects of L-ascorbate (vitamin C, Vit. C) on METH toxicity and HO-1 expression in neuronal/glial cocultures. Cell viability and damage were evaluated by 3-(4,5-dimethylthianol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) reduction and lactate dehydrogenase (LDH) release, respectively. Neuronal and glial localization of HO-1 were identified by double immunofluorescence staining. Reactive oxygen species (ROS) production was measured using the fluorochrome 2′,7′-dichlorofluorescin diacetate. HO-1 mRNA and protein expression were examined by RT-qPCR and Western blotting, respectively. Results show that Vit. C induced HO-1 mRNA and protein expressions in time- and concentration-dependent manners. Inhibition of p38 mitogen-activated protein kinase (MAPK) but not extracellular signal-regulated kinase (ERK) significantly blocked induction of HO-1 by Vit. C. HO-1 mRNA and protein expressions were significantly elevated by a combination of Vit. C and METH, compared to either Vit. C or METH alone. Pretreatment with Vit. C enhanced METH-induced HO-1 expression and attenuated METH-induced ROS production and neurotoxicity. Pharmacological inhibition of HO activity abolished suppressive effects of Vit. C on METH-induced ROS production and attenuated neurotoxicity. We conclude that induction of HO-1 expression contributes to the attenuation of METH-induced ROS production and neurotoxicity by Vit. C. We suggest that HO-1 induction by Vit. C may serve as a strategy to alleviate METH neurotoxicity. -- Highlights: ► Besides the anti-oxidant effect, Vit. C also induces HO-1 expression in brain cells. ► Vit. C reduces METH neurotoxicity and ROS production by

  8. Catalytic nanomedicine technology: copper complexes loaded on titania nanomaterials as cytotoxic agents of cancer cell

    PubMed Central

    Lopez, Tessy; Ortiz-Islas, Emma; Guevara, Patricia; Gómez, Esteban

    2013-01-01

    The anticancer properties of pure copper (II) acetate and copper (II) acetylacetonate, alone and loaded on functionalized sol-gel titania (TiO2), were determined in four different cancer cell lines (C6, RG2, B16, and U373), using increasing concentrations of these compounds. The copper complexes were loaded onto the TiO2 network during its preparation by the solgel process. Once copper-TiO2 materials were obtained, these were characterized by several physical-chemical techniques. An in vitro copper complex–release test was developed in an aqueous medium at room temperature and monitored by ultraviolet spectroscopy. The toxic effect of the copper complexes, alone and loaded on TiO2, was determined using a cell viability 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, when cancer cells were treated with increasing concentrations (15.75–1000 mg/mL) of these. Characterization studies revealed that the addition of copper complexes to the TiO2 sol-gel network during its preparation, did not generate changes in the molecular structure of the complexes. The surface area, pore volume, and pore diameter were affected by the copper complex additions and by the crystalline phases obtained. The kinetic profiles of both copper complexes released indicated two different stages of release: The first one was governed by first-order kinetics and the second was governed by zero-order kinetics. The cell viability assay revealed a cytotoxic effect of copper complexes, copper-TiO2, and cisplatin in a dose-dependent response for all the cell lines; however, the copper complexes exhibited a better cytotoxic effect than the cisplatin compound. TiO2 alone presented a minor cytotoxicity for C6 and B16 cells; however, it did not cause any toxic effect on the RG2 and U373 cells, which indicates its high biocompatibility with these cells. PMID:23413123

  9. The viability of mouse spermatogonial germ cells on a novel scaffold, containing human serum albumin and calcium phosphate nanoparticles

    PubMed Central

    Yadegar, Mona; Hekmatimoghaddam, Seyed Hossein; Nezami Saridar, Saeide; Jebali, Ali

    2015-01-01

    Background: In spermatogenesis, spermatogonial cells differentiate to the haploid gametes. It has been shown that spermatogenesis can be done at in vitro condition. In vitro spermatogenesis may provide an open window to treat male infertility. Objective: The aim of this study was to evaluate the effects of a novel scaffold containing human serum albumin (HSA)/tri calcium phosphate nanoparticles (TCP NPs) on the mouse spermatogonial cell line (SCL). Materials and Methods: First, TCP NPs were synthesized by reaction of calcium nitrate and diammonium phosphate at pH 13. Then, serial concentrations of TCP NPs were separately added to 500 mg/mL HSA, and incubated in the 100oC water for 30 min. In the next step, each scaffold was cut (2×2mm), placed into sterile well of microplate, and then incubated for 1, 2, and 3 days at 37oC with mouse SCL. After incubation, the cytotoxicity of the scaffolds was evaluated by different tests including 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) assay, vital staining, and cell counting. On the other hand, the release of TCP NPs and HSA from the scaffolds was measured. Results: Based on microscopic observation, the size of cavities for all scaffolds was near 200-500 µm, and the size of TCP NPs was near 50-100 nm. All toxicity tests showed that the increase of TCP concentration in the scaffold did not affect mouse SCL. It means that the percentage of cell viability, LDH release, vital cells, and cell quantity was 85%, 105%, 90%, and 110%, respectively. But, the increase of incubation time led to increase of LDH release (up to 115%) and cell count (up to 115%). Also, little decrease of cell viability and vital cells was seen when incubation time was increased. Here, no release of TCP NPs and HSA was seen after increase of TCP concentration and incubation time. Conclusion: It can be concluded that the increase of TCP concentration in HSA/ TCP NPs scaffold does not lead to

  10. Potent Anticancer Effects of Bioactive Mushroom Extracts (Phellinus linteus) on a Variety of Human Cancer Cells

    PubMed Central

    Konno, Sensuke; Chu, Kevin; Feuer, Nicholas; Phillips, John; Choudhury, Muhammad

    2015-01-01

    Background Although several therapeutic options are currently available for patients with various cancers, the outcomes are often disappointing and a more effective modality needs to be promptly established. We have been exploring an alternative approach using natural agents and two bioactive mushroom extracts isolated from Phellinus linteus (PL), namely PL-ES and PL-I-ES, were of our interest. As anticancer effects of similar extracts have been reported in several cancers, we investigated whether PL-ES and PL-I-ES might have such anticancer activities on a variety of human cancer cells in vitro. Methods Ten different types of human cancer cell lines, including three metastatic prostate, bladder, kidney, lung, breast, stomach, liver, and brain cancer cells, were employed and tested with PL-ES or PL-I-ES. Cell growth/viability, exertion of oxidative stress, and induction of apoptosis were assessed by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay, lipid peroxidation (LPO) assay, and specific enzymatic assay, respectively. Results PL-ES (100 µg/mL) exhibited potent anticancer activity, resulting in a significant (40-80%) growth reduction in all 10 cancer cells at 72 hours. PL-I-ES (100 µg/mL) was effective on only four cancer cells but its higher concentration at 250 µg/mL led to a significant (25-90%) growth reduction in seven cancer cells. LPO assays indicated that such a significant growth reduction by PL-ES (100 µg/mL) or PL-I-ES (100 or 250 µg/mL) could result from cell death due to a cytotoxic effect of oxidative stress (through free radicals). Moreover, enzymatic assays for caspase-3 (Csp-3) and caspase-9 (Csp-9), the pro-apoptotic regulators, showed that both enzymes were significantly activated by PL-ES or PL-I-ES, indicating that cell death due to oxidative stress was more likely associated with apoptosis. Conclusions The present study shows that both PL-ES and PL-I-ES indeed have anticancer effects on a variety of cancer

  11. Chemical Composition and In Vitro Cytotoxicity of Essential Oils from Leaves and Flowers of Callistemon citrinus from Western Himalayas

    PubMed Central

    Kumar, Dharmesh; Sukapaka, Mahesh; Babu, G. D. Kiran; Padwad, Yogendra

    2015-01-01

    Background Plant-based traditional system of medicine continues to play an important role in healthcare. In order to find new potent source of bioactive molecules, we studied the cytotoxic activity of the essential oils from the flowers and leaves of Callistemon citrinus. This is the first report on anticancer potential of essential oils of C. citrinus. Methods Cytotoxicity of essential oil was evaluated using sulfo-rhodamine B (SRB) assay against human lung carcinoma (A549), rat glioma (C-6), human colon cancer (Colo-205) and human cervical cancer (SiHa) cells. Apoptosis induction was evaluated by caspase-3/7 activity which was further confirmed by western blotting. Percentage cell apoptosis was determined by Annexin V based dead cell assay followed by DNA content as cell cycle analysis against A549 and C-6 cells. While 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to check the toxicity against normal human peripheral blood mononuclear cells (PBMCs), the immunomodulatory activity on mouse splenocytes was evaluated using SRB assay. Results The GC and GC-MS analysis of these essential oils revealed high content of α-pinene (32.3%), limonene (13.1%) and α-terpineol (14.6%) in leaf sample, whereas the flower oil was dominated by 1,8-cineole (36.6%) followed by α-pinene (29.7%). The leaf oil contained higher amount of monoterpene hydrocarbons (52.1%) and sesquiterpenoids (14%) as compared to flower oil (44.6% and 1.2%, respectively). However, the flower oil was predominant in oxygenated monoterpenes (43.5%). Although both leaf and flower oils showed highest cytotoxicity on A549 cells (61.4%±5.0 and 66.7%±2.2, respectively), only 100 μg/mL flower oil was significantly active against C-6 cells (69.1%±3.1). Interestingly, no toxicity was recorded on normal cells. Conclusion Higher concentration of 1,8-cineole and/or synergistic effect of the overall composition were probably responsible for the efficacy of flower and leaf oils

  12. A Novel Hsp90 Inhibitor Activates Compensatory Heat Shock Protein Responses and Autophagy and Alleviates Mutant A53T α-Synuclein Toxicity

    PubMed Central

    Xiong, Rui; Zhou, Wenbo; Siegel, David; Kitson, Russell R. A.; Freed, Curt R.; Moody, Christopher J.

    2015-01-01

    A potential cause of neurodegenerative diseases, including Parkinson’s disease (PD), is protein misfolding and aggregation that in turn leads to neurotoxicity. Targeting Hsp90 is an attractive strategy to halt neurodegenerative diseases, and benzoquinone ansamycin (BQA) Hsp90 inhibitors such as geldanamycin (GA) and 17-(allylamino)-17-demethoxygeldanamycin have been shown to be beneficial in mutant A53T α-synuclein PD models. However, current BQA inhibitors result in off-target toxicities via redox cycling and/or arylation of nucleophiles at the C19 position. We developed novel 19-substituted BQA (19BQA) as a means to prevent arylation. In this study, our data demonstrated that 19-phenyl-GA, a lead 19BQA in the GA series, was redox stable and exhibited little toxicity relative to its parent quinone GA in human dopaminergic SH-SY5Y cells as examined by oxygen consumption, trypan blue, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), and apoptosis assays. Meanwhile, 19-phenyl-GA retained the ability to induce autophagy and potentially protective heat shock proteins (HSPs) such as Hsp70 and Hsp27. We found that transduction of A53T, but not wild type (WT) α-synuclein, induced toxicity in SH-SY5Y cells. 19-Phenyl-GA decreased oligomer formation and toxicity of A53T α-synuclein in transduced cells. Mechanistic studies indicated that mammalian target of rapamycin (mTOR)/p70 ribosomal S6 kinase signaling was activated by A53T but not WT α-synuclein, and 19-phenyl-GA decreased mTOR activation that may be associated with A53T α-synuclein toxicity. In summary, our results indicate that 19BQAs such as 19-phenyl-GA may provide a means to modulate protein-handling systems including HSPs and autophagy, thereby reducing the aggregation and toxicity of proteins such as mutant A53T α-synuclein. PMID:26405178

  13. The Effect of Artemisia fragrans Willd: Essential Oil on Inducible Nitric Oxide Synthase Gene Expression and Nitric Oxide Production in Lipopolysaccharide-stimulated Murine Macrophage Cell Line.

    PubMed

    Farghadan, Maryam; Ghafoori, Hosein; Vakhshiteh, Faezeh; Shahzadeh Fazeli, Seyed Abolhassan; Farzaneh, Parvaneh; Kokhaei, Parviz

    2016-12-01

    The genus Artemisia is estimated to comprise over 800 species with anti-cancer, anti-fungal, anti-oxidant and anti-inflammatory properties. Artemisia fragrans (A. fragrans), a species that belongs to genus Artemisia, is rich in monoterpenes and sesquiterpenes derivatives. Due to anti-inflammatory properties of monoterpenes and sesquiterpenes, we aimed to investigate the effect of A. fragrans essential oil on mRNA expression of inducible nitric oxide synthase (iNOS) gene and nitric oxide (NO) production in Lipopolysaccharide (LPS) -stimulated RAW264.7 cell line. NO, which is synthesized by iNOS, is the main macrophage-derived inflammatory mediator. The oil obtained from the A. fragrans was prepared from aerial parts of the plant. Chemical composition of essential oil was analyzed by gas chromatography-mass spectrometry (GC/MS).The cytotoxicity of various concentrations of essential oil was evaluated by mitochondrial reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test assay. The effect of different doses (1.75-7 mg/mL) of A. fragrans oil on mRNA expression of iNOS gene and NO production in LPS-stimulated RAW 264.7 cells was assessed by real-time PCR method and Griess reagent, respectively. In GC/MS analyses of A. fragrans oil, 32 compounds were identified. The main components of the oil were camphor and 1, 8-cineole. The results demonstrated that the essential oil of A. fragrans (1.75- 7 mg/mL), in a dose-dependent manner, inhibits mRNA expression of iNOS induced by LPS in the RAW264.7 cells without cytotoxic effect even at higher doses. The results of iNOS were consistent with the results of NO production. Our preliminary results suggest the possible anti-inflammatory effect of A. fragrans. Further studies are needed to determine the full pharmacokinetics of A. fragrans activity in vivo.

  14. Endocannabinoids and inflammatory response in periodontal ligament cells.

    PubMed

    Özdemir, Burcu; Shi, Bin; Bantleon, Hans Peter; Moritz, Andreas; Rausch-Fan, Xiaohui; Andrukhov, Oleh

    2014-01-01

    Endocannabinoids are associated with multiple regulatory functions in several tissues. The main endocannabinoids, anandamide (AEA) and 2-arachidonylglycerol (2-AG), have been detected in the gingival crevicular fluid of periodontitis patients, but the association between periodontal disease or human periodontal ligament cells (hPdLCs) and endocannabinoids still remain unclear. The aim of the present study was to examine the effects of AEA and 2-AG on the proliferation/viability and cytokine/chemokine production of hPdLCs in the presence/absence of Porphyromonas gingivalis lipopolysaccharide (P. gingivalis LPS). The proliferation/viability of hPdLCs was measured using 3,4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT)-assay. Interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemotactic protein-1 (MCP-1) levels were examined at gene expression and protein level by real-time PCR and ELISA, respectively. AEA and 2-AG did not reveal any significant effects on proliferation/viability of hPdLCs in the absence of P. gingivalis LPS. However, hPdLCs viability was significantly increased by 10-20 µM AEA in the presence of P. gingivalis LPS (1 µg/ml). In the absence of P. gingivalis LPS, AEA and 2-AG did not exhibit any significant effect on the expression of IL-8 and MCP-1 expression in hPdLCs, whereas IL-6 expression was slightly enhanced by 10 µM 2-AG and not affected by AEA. In P.gingivalis LPS stimulated hPdLCs, 10 µM AEA down-regulated gene-expression and protein production of IL-6, IL-8, and MCP-1. In contrast, 10 µM 2-AG had an opposite effect and induced a significant up-regulation of gene and protein expression of IL-6 and IL-8 (P<0.05) as well as gene-expression of MCP-1 in P. gingivalis LPS stimulated hPdLCs. Our data suggest that AEA appears to have an anti-inflammatory and immune suppressive effect on hPdLCs' host response to P.gingivalis LPS, whereas 2-AG appears to promote detrimental inflammatory processes. In conclusion, AEA and 2

  15. A combinatorial approach of N-terminus blocking and codon optimization strategies to enhance the soluble expression of recombinant hIL-7 in E. coli fed-batch culture.

    PubMed

    Devi, Nirmala; Adivitiya; Khasa, Yogender Pal

    2016-12-01

    Human interleukin-7 (hIL-7) is a therapeutically important cytokine involved in lymphocyte development and survival. In previous reports, a uniformly poor expression of hIL-7 has been shown in Escherichia coli host with the problem of inclusion body formation. In this study, the role of codon optimization and N-terminus blocking using various solubility enhancer fusion tags was explored to improve its soluble expression. The use of codon optimization strategy improved its expression to 80 ± 5 mg/L at shake flask level. The utilization of pelB leader sequence resulted in an unprocessed protein in the form of cytoplasmic inclusion bodies with lower expression yields. The N-terminus fusion of small ubiquitin-like modifier (SUMO), thioredoxin (Trx), and NusA tags increased the expression in the range of 90-140 mg/L, where >90 % of the fusion protein was obtained in soluble form. The fed-batch fermentation of SUMO-tagged hIL-7 protein was optimized at bioreactor level, where a high volumetric product concentration of 2.65 g/L was achieved by controlling the plasmid segregation instability using high antibiotic concentration. The specific product yield (YP/X) and volumetric product concentration were 1.38 and 2.55-fold higher compared to batch results, respectively. A preparative scale affinity chromatography resulted in a high recovery yield of 50.6 mg/L with ∼90 % purity. The conformational property of purified recombinant hIL-7 from CD spectroscopy showed a typical helical structure with 31.5 % α-helix and 26.43 % β-sheet. The biological activity of purified protein was tested using IL-7-dependent murine immature B lymphocyte (2E8) cell line by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide salt (MTT) assay, where it showed a similar biological activity as standard control.

  16. Activation of Type 4 Metabotropic Glutamate Receptor Attenuates Oxidative Stress-Induced Death of Neural Stem Cells with Inhibition of JNK and p38 MAPK Signaling

    PubMed Central

    Zhang, Zhichao; Ma, Wen; Wang, Li; Gong, Hanshi; Tian, Yumei; Zhang, Jianshui; Liu, Jianxin; Lu, Haixia

    2015-01-01

    Promoting both endogenous and exogenous neural stem cells' (NSCs) survival in the hostile host environments is essential to cell replacement therapy for central nervous system (CNS) disorders. Type 4 metabotropic glutamate receptor (mGluR4), one of the members of mGluRs, has been shown to protect neurons from acute and chronic excitotoxic insults in various brain damages. The present study investigated the preventive effects of mGluR4 on NSC injury induced by oxidative stress. Under challenge with H2O2, loss of cell viability was observed in cultured rat NSCs, and treatment with selective mGluR4 agonist VU0155041 conferred protective effects against the loss of cellular viability in a concentration-dependent manner, as shown by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Pretreatment of VU0155041 (30 μM) also inhibited the excessive NSC death induced by H2O2, and group III mGluRs antagonist (RS)-a-methylserine-O-phosphate (MSOP) or gene-targeted knockdown abolished the protective action of mGluR4, indicated by propidium iodide–Hoechst and terminal deoxynucleotidyl transferase-mediated UTP nick end labeling (TUNEL) staining. Western blot assay demonstrated that mGluR4 activation reversed the decreased procaspase-8/9/3and the destructed Bcl-2/Bax expressing balance, and likewise, MSOP and mGluR4 knockdown abrogated the action of mGluR4 activity. Furthermore, inhibition of JNK and p38 mitogen-activated protein kinases (MAPKs) were observed after mGluR4 activation, and as paralleling control, JNK-specific inhibitor SP600125 and p38-specific inhibitor SB203580 significantly rescued the H2O2-mediated NSC apoptosis and cleavage of procaspase-3. We suggest that activation of mGluR4 prevents oxidative stress-induced NSC death and apoptotic-associated protein activities with involvement of inhibiting the JNK and p38 pathways in cell culture. Our findings may help to develop strategies for enhancing the resided and transplanted NSC survival

  17. ZnO nanoparticles induced oxidative stress and apoptosis in HepG2 and MCF-7 cancer cells and their antibacterial activity.

    PubMed

    Wahab, Rizwan; Siddiqui, Maqsood A; Saquib, Quaiser; Dwivedi, Sourabh; Ahmad, Javed; Musarrat, Javed; Al-Khedhairy, Abdulaziz A; Shin, Hyung-Shik

    2014-05-01

    Liver and breast cancer are the most traumatic diseases because they affect the major organs of the body. Nanomedicine recently emerged as a better option for the treatment of these deadly diseases. As a result, many nanoparticles have been used to treat cancer cell lines. Of the various nanoparticles, zinc oxide exhibits biocompatibility. Therefore, the aim of the present study was to investigate the activity of zinc oxide nanoparticles (ZnO-NPs) against HepG2 and MCF-7 cells. The NPs (∼13±2 nm) were prepared via a non-protonated chemical route and were well-characterized through standard techniques. The study showed that treatment with NPs is notably effective against the proliferation of HepG2 and MCF-7 cancer cells in a dose-dependent manner. The MTT (3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide, a tetrazole) assays revealed the concentration-dependent cytotoxic effects of NPs in range of 2.5-100 μg/ml. HepG2 and MCF-7 cells were exposed to ZnO-NPs and exhibited a significant reduction in their cell viability (95% and 96%; p<0.05) in response to a very low concentration (25 μg/ml) of the ZnO-NPs; this finding was confirmed with FACS (fluorescence-activated cell sorting) data. The reduction in cell viability in response to NP treatment induces cytotoxicity in the cultured cells. The quantitative RT-PCR (real-time polymerase chain reaction) results demonstrate that the exposure of HepG2 cells to ZnO-NPs results in significant upregulation of the mRNA expression level of Bax, p53, and caspase-3 and the down regulation of the anti-apoptotic gene Bcl-2. The NPs were also tested against five pathogenic bacteria through the disk diffusion method, and their antibacterial activities were compared with that of ZnO salt.

  18. Investigation of functional and morphological integrity of freshly isolated and cryopreserved human hepatocytes.

    PubMed

    Ostrowska, A; Bode, D C; Pruss, J; Bilir, B; Smith, G D; Zeisloft, S

    2000-01-01

    There is a pressing need for alternative therapeutic methods effective in the treatment of patients with liver insufficiency. Isolated human hepatocytes may be a viable alternative or adjunct to orthotopic liver transplantation in such patients. The purpose of this study was to evaluate the viability and functional integrity of freshly isolated and cryopreserved human hepatocytes, in preparation for a multi-center human hepatocyte transplantation trial. We are currently processing transplant-grade human parenchymal liver cells from nondiseased human livers that are obtained through a network of organ procurement organizations (OPOs). Thus far, sixteen hepatocyte transplants have been performed using hepatocytes processed by our methods. At the time of referral all specimens were deemed unsuitable for transplantation due to anatomical anomalies, high fat content, medical history, etc. Hepatocytes were isolated from encapsulated liver sections by a modified two-step perfusion technique. Isolated cells were cryopreserved and stored in liquid nitrogen for one to twelve months. The total yield of freshly isolated hepatocytes averaged 3.7x10(7) cells per gram of wet tissue. Based on trypan blue exclusion, fresh preparations contained an average of 85% viable hepatocytes vs. 70% in cryopreserved samples. The plating efficiencies of cells seeded immediately after isolation ranged from 87% to 98%, while those of cryopreserved/thawed cells were markedly lower. Flow cytometry analysis of cells labeled with 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) showed that there was no significant difference in viability compared with trypan blue staining. Both freshly isolated hepatocytes and those recovered from cryopreservation showed typical and intact morphology as demonstrated by light and electron microscopy. The product of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reaction was always expressed more intensely in cultures of freshly

  19. Toxicity of nano- and micro-sized silver particles in human hepatocyte cell line L02

    NASA Astrophysics Data System (ADS)

    Liu, Pengpeng; Guan, Rongfa; Ye, Xingqian; Jiang, Jiaxin; Liu, Mingqi; Huang, Guangrong; Chen, Xiaoting

    2011-07-01

    Silver nanoparticles (Ag NPs) previously classified as antimicrobial agents have been widely used in consumers and industrial products, especially food storage material. Ag NPs used as antimicrobial agents may be found in liver. Thus, examination of the ability of Ag NPs to penetrate the liver is warranted. The aim of the study was to determine the optimal viability assay for using with Ag NPs in order to assess their toxicity to liver cells. For toxicity evaluations, cellular morphology, mitochondrial function (3-(4, 5-dimethylazol-2-yl)-2, 5-diphenyl-tetrazolium bromide, MTT assay), membrane leakage of lactate dehydrogenase (lactate dehydrogenase, LDH release assay), Oxidative stress markers (malonaldehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD)), DNA damage (single cell gel eletrophoresis, SCGE assay), and protein damage were assessed under control and exposed conditions (24 h of exposure). The results showed that mitochondrial function decreased significantly in cells exposed to Ag NPs at 25 μg·mL-1. LDH leakage significantly increased in cells exposed to Ag NPs (>= 25 μg mL-1) while micro-sized silver particles tested displayed LDH leakage only at higher doses (100 μg·mL-1). The microscopic studies demonstrated that nanoparticle-exposed cells at higher doses became abnormal in size, displaying cellular shrinkage, and an acquisition of an irregular shape. Due to toxicity of silver, further study conducted with reference to its oxidative stress. The results exhibited significant depletion of GSH level, increase in SOD levels and lead to lipid peroxidation, which suggested that cytotoxicity of Ag NPs in liver cells might be mediated through oxidative stress. The results demonstrates that Ag NPs lead to cellular morphological modifications, LDH leakage, mitochondrial dysfunction, and cause increased generation of ROS, depletion of GSH, lipid peroxidation, oxidative DNA damage and protein damage. Though the exact mechanism behind Ag NPs

  20. Tissue factor induces VEGF expression via activation of the Wnt/β-catenin signaling pathway in ARPE-19 cells

    PubMed Central

    Wang, Ying; Sang, Aimin; Zhu, Manhui; Zhang, Guowei; Guan, Huaijin; Ji, Min

    2016-01-01

    Purpose The purpose of the present study was to investigate the potential signal mechanism of tissue factor (TF) in the regulation of the expression of vascular endothelial growth factor (VEGF) in human retinal pigment epithelial (ARPE-19) cells. Methods An in vitro RPE cell chemical hypoxia model was established by adding cobalt chloride (CoCl2) in the culture medium. The irritative concentration of CoCl2 was determined with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay kit. VEGF production in ARPE-19 cells was measured with enzyme-linked immunosorbent assay (ELISA) and western blotting. The Wnt signaling pathway–associated molecules, including phospho-glycogen synthase kinase 3β (p-GSK3β), GSK3β, p-β-catenin and β-catenin, were detected with western blotting. pEGFP-N3-hTF was constructed and verified with digestion of the restriction enzyme and sequencing analysis. Human TF overexpression and silencing plasmids were transfected into the ARPE-19 cells to clarify the causal relationship between TF and VEGF expression. The Transwell coculture system of ARPE-19 cells and RF/6A rhesus macaque choroid–retinal endothelial cells was performed to evaluate cell invasion and tube formation ability. Results Our anoxic model of ARPE-19 cells showed that TF expression was upregulated in accordance with variations in hypoxia-inducible factor 1-alpha (HIF-1α) and VEGF levels. Silencing and overexpression of TF decreased and increased VEGF expression, respectively. The Wnt/β-catenin signaling pathway played an important role in this effect. Results from the ARPE-19 cell and RF/6A cell coculture system showed that the enhancement of TF expression in the ARPE-19 cells led to significantly faster invasion and stronger tube-forming ability of the RF/6A cells, while siRNA-mediated TF silencing caused the opposite effects. Pharmacological disruption of Wnt signaling IWR-1-endo inhibited the effects compared to the TF-overexpressing group

  1. Glucagon like peptide-2 induces intestinal restitution through VEGF release from subepithelial myofibroblasts.

    PubMed

    Bulut, Kerem; Pennartz, Christian; Felderbauer, Peter; Meier, Juris J; Banasch, Matthias; Bulut, Daniel; Schmitz, Frank; Schmidt, Wolfgang E; Hoffmann, Peter

    2008-01-14

    Glucagon like peptide-2 (GLP-2) exerts intestinotrophic actions, but the underlying mechanisms are still a matter of debate. Recent studies demonstrated the expression of the GLP-2 receptor on fibroblasts located in the subepithelial tissue, where it might induce the release of growth factors such as keratinocyte growth factor (KGF) or vascular endothelial growth factor (VEGF). Therefore, in the present studies we sought to elucidate the downstream mechanisms involved in improved intestinal adaptation by GLP-2. Human colonic fibroblasts (CCD-18Co), human colonic cancer cells (Caco-2 cells) and rat ileum IEC-18 cells were used. GLP-2 receptor mRNA expression was determined using real time RT-PCR. Conditioned media from CCD-18Co cells were obtained following incubation with GLP-2 (50-250 nM) for 24 h. Cell viability was assessed by a 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium bromide (MTT)-assay, and wound healing was determined with an established migration-assay. Transforming Growth Factor beta (TGF-beta), VEGF and KGF mRNA levels were determined by RT-PCR. Protein levels of VEGF and TGF-beta in CCD-18Co cells following GLP-2 stimulation were determined using ELISA. Neutralizing TGF-beta and VEGF-A antibodies were utilized to assess the role of TGF-beta and VEGF-A in the process of wound healing. GLP-2 receptor expression was detected in CCD-18Co cells. Conditioned media from CCD-18Co cells dose-dependently induced proliferation in Caco-2 cells, but not in IEC-18 cells. Conditioned media also enhanced cell migration in IEC-18 cells (P<0.01), while migration was even inhibited in Caco-2 cells (P<0.0012). GLP-2 significantly stimulated mRNA expression of VEGF and TGF-beta, but not of KGF in CCD-18Co. The migratory effects of GLP-2 were completely abolished in the presence of TGF-beta and VEGF-A antibodies. GLP-2 exerts differential effects on the epithelium of the small intestine and the colon. Thus, in small intestinal cells GLP-2 stimulates wound

  2. Inhibition of ERK1/2 Worsens Intestinal Ischemia/Reperfusion Injury

    PubMed Central

    Ban, Kechen; Peng, Zhanglong; Kozar, Rosemary A.

    2013-01-01

    Background The role of extracellular signal-regulated protein kinase (ERK) in intestinal ischemia/reperfusion (I/R) injury has not been well investigated. The aim of the current study was to examine the effect of inhibition of the ERK pathway in an in vitro and in vivo model of intestinal I/R injury. Methods ERK1/2 activity was inhibited using the specific inhibitor, U0126, in intestinal epithelial cells under hypoxia/reoxygenation conditions and in mice subjected to 1 hour of intestinal ischemia followed by 6 hours reperfusion. In vitro, cell proliferation was assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, apoptosis by DNA fragmentation, and migration using an in vitro model of intestinal wound healing. Cells were also transfected with a p70S6K plasmid and the effects of overexpression similarly analyzed. In vivo, the effects of U0126 on intestinal cell proliferation and apoptosis, intestinal permeability, lung and intestinal neutrophil infiltration and injury, and plasma cytokine levels were measured. Survival was also assessed after U0126. Activity of p70S6 kinase (p70S6K) was measured by Western blot. Results In vitro, inhibition of ERK1/2 by U0126 significantly decreased cell proliferation and migration but enhanced cell apoptosis. Overexpression of p70S6K promoted cell proliferation and decreased cell apoptosis. In vivo, U0126 significantly increased cell apoptosis and decreased cell proliferation in the intestine, increased intestinal permeability, intestinal and lung neutrophil infiltration, and injury, as well as systemic pro-inflammatory cytokines, TNF-α, IL-6 and IL-1β. Mortality was also significantly increased by U0126. Inhibition of ERK1/2 by U0126 also abolished activity of p70S6K both in vitro and in vivo models. Conclusion Pharmacologic inhibition of ERK1/2 by U0126 worsens intestinal IR injury. The detrimental effects are mediated, at least in part, by inhibition of p70S6K, the major effector of mammalian

  3. Evaluation of the antibacterial and anticancer activities of some South African medicinal plants

    PubMed Central

    2011-01-01

    Background Several herbs are traditionally used in the treatment of a variety of ailments particularly in the rural areas of South Africa where herbal medicine is mainly the source of health care system. Many of these herbs have not been assessed for safety or toxicity to tissue or organs of the mammalian recipients. Methods This study evaluated the cytotoxicity of some medicinal plants used, inter alia, in the treatment of diarrhoea, and stomach disorders. Six selected medicinal plants were assessed for their antibacterial activities against ampicillin-resistant and kanamycin-resistant strains of Escherichia coli by the broth micro-dilution methods. The cytotoxicities of methanol extracts and fractions of the six selected plants were determined using a modified tetrazolium-based colorimetric assay (3-(4, 5-dimethylthiazol)-2, 5-diphenyl tetrazolium bromide (MTT) assay). Results The average minimum inhibitory concentration (MIC) values of the plants extracts ranged from 0.027 mg/mℓ to 2.5 mg/mℓ after 24 h of incubation. Eucomis autumnalis and Cyathula uncinulata had the most significant biological activity with the least MIC values. The in vitro cytotoxicity assay on human hepatocarcinoma cell line (Huh-7) revealed that the methanol extract of E. autumnalis had the strongest cytotoxicity with IC50 of 7.8 μg/mℓ. Ethyl acetate and butanol fractions of C. uncinulata, Hypoxis latifolia, E. autumnalis and Lantana camara had lower cytotoxic effects on the cancer cell lines tested with IC50 values ranging from 24.8 to 44.1 μg/mℓ; while all the fractions of Aloe arborescens and A. striatula had insignificant or no cytotoxic effects after 72 h of treatment. Conclusions Our results indicate that the methanol fraction of E. autumnalis had a profound cytotoxic effect even though it possessed very significant antibacterial activity. This puts a query on its safety and hence a call for caution in its usage, thus a product being natural is not tantamount to being

  4. Chemical alternatives to methyl bromide for Florida ornamental production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This project is a cooperative effort among USDA, ARS and University of Florida researchers, Florida in-ground ornamental producers, and fumigant industry representatives. Funding is provided through the USDA-ARS Area-wide Pest Management Program for Alternatives to Methyl Bromide. The ornamental i...

  5. Status of Alternatives for Methyl Bromide in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methyl bromide is a fumigant used for disinfestation of soils, commodities and structures. Listed as an ozone-depleting chemical international environmental protocols and the U.S. Clean Air Act require that its use be severely restricted. Although use of this fumigant has fallen considerably, the U....

  6. EFFECT OF BROMIDE ION ON FORMATION OF HAAS DURING CHLORINATION

    EPA Science Inventory

    loacetic acids (HAAs) during chlorination and he effects of independent variables, including pH, reaction time, and chlorine dosage. Almost all of the indpendent loaetic acids (HAAs) during chlorin...designed to statistically evaluate the influence of bromide ion on the formatio...

  7. Depleting methyl bromide residues in soil by reaction with bases

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Despite generally being considered the most effective soil fumigant, methyl bromide (MeBr) use is being phased out because its emissions from soil can lead to stratospheric ozone depletion. However, a large amount is still currently used due to Critical Use Exemptions. As strategies for reducing the...

  8. REVIEW OF CONTROL OPTIONS FOR METHYL BROMIDE IN COMMODITY TREATMENT

    EPA Science Inventory

    The report describes recent developments in the control of methyl bromide (MeBr) and discusses technical considerations and requirements for and economic feasibility of recovery. (NOTE: MeBr, fumigant for agricultural commodities, is an ozone depleting chemical. The U.S. EPA has ...

  9. Evaluation of the Efficacy of Methyl Bromide in the ...

    EPA Pesticide Factsheets

    Journal Article The objective of this article is to determine the required conditions for the effective inactivation of Bacillus anthracis (B.a.) spores on materials using methyl bromide (MeBr) and to obtain comparative efficacy data with three avirulent microorganisms, to assess their potential as surrogates for B.a. Ames.

  10. The Fate of Alternative Soil Funigants to Methyl Bromide

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Soil fumigation is an important agricultural practice for the control of soil-borne pests. Since the phase–out of methyl bromide, due to its role in the depletion of stratospheric ozone, several alternatives such as 1,3-dichloropropene (1,3-D), chloropicrin (CP), and dimethyl disulfide (DMDS) are b...

  11. Ipratropium bromide spray as treatment for sialorrhea in Parkinson's disease.

    PubMed

    Thomsen, Teri R; Galpern, Wendy R; Asante, Abena; Arenovich, Tamara; Fox, Susan H

    2007-11-15

    Sialorrhea is a significant problem in advanced Parkinson's disease (PD). Current treatment options include systemic anticholinergics which frequently cause side effects. We hypothesized that sublingual application of ipratropium bromide spray, an anticholinergic agent that does not cross the blood brain barrier, may reduce drooling without systemic side effects. We performed a randomized, double blind, placebo-controlled, crossover study in 17 subjects with PD and bothersome drooling. Patients were randomized to receive ipratropium bromide or placebo (one to two sprays, maximum of four times per day) for 2 weeks followed by a 1 week washout and crossover for further 2 weeks of treatment. The primary outcome was an objective measure of weight of saliva production. Secondary outcomes were subjective rating of severity and frequency of sialorrhoea using home diaries, United Parkinson's Disease Rating Scale (UPDRS) part II salivation subscore, parkinsonian disability using UPDRS, and adverse events. Ipratropium bromide spray had no significant effect on weight of saliva produced. There was a mild effect of treatment on subjective measures of sialorrhea. There were no significant adverse events. Ipratropium bromide spray was well tolerated in subjects with PD. Although it did not affect objective measures of saliva production, further studies in parkinsonism may be warranted.

  12. Actual hazard of methyl bromide fumigation in soil disinfection.

    PubMed Central

    Van Den Oever, R U; Roosels, D; Lahaye, D

    1982-01-01

    Methyl bromide, a highly toxic and ready penetrating fumigant, is widely used against rodents, insects, mites, and a range of pathogenic organisms in soil, compost, and timber. To disinfect soil in greenhouses, methyl bromide is brought under pressure from outside by a vaporiser and blown on to ground under a polyethylene cover. The gas being three times heavier than air easily penetrates the ground. Depending on the local ventilation, a considerable amount of gas evaporates into the surrounding atmosphere, this emission being especially serious during the fumigation procedure and at the removal of the plastic cover. Previously, mechanical injection of methyl bromide on to the ground within closed areas was prohibited, since this technique exposed at least four disinfection workers at a time, who were provided with only a canister respirator, to gas concentrations of over 1000 ppm CH3Br. The present study established that fumigation with methyl bromide also carries risks for the well-protected worker inside, as well as for the one controlling the vaporiser. The concentration during application varies from 30 to 3000 ppm. Concentration in the air declines with time to 4 ppm CH3Br five days after application. Discarding the plastic sheet involves exposure to peak values as high as 200 ppm for a few seconds. On the ninth day after application, milling the soil can expose workers to up to 15 ppm; on the eleventh day no CH3Br concentration in the air could be found. PMID:7066229

  13. Bromide ion effect on N-nitrosodimethylamine formation by monochloramine.

    PubMed

    Luh, Jeanne; Mariñas, Benito J

    2012-05-01

    N-Nitrosodimethylamine (NDMA) formation experiments conducted in phosphate buffer demonstrated that in waters containing monochloramine, the presence of bromide ion enhanced NDMA formation at the relatively high pH values of 8 and 9 after 24 h of reaction time, which was consistent with literature results. However, at relatively low to neutral pH (6 to 7), the presence of bromide resulted in lower NDMA formation as compared to results obtained in the absence of bromide. The hypothesis that bromamines were the species directly responsible for enhanced NDMA formation at high pH was tested and was shown not to be valid. Additional active bromine species were also tested, including hypobromous acid, hypobromite ion, and tribromide ion, with no species showing an ability to directly enhance NDMA formation. Analysis of the UV spectral data corresponding to the NDMA experiments suggest that the mechanism by which bromide enhances NDMA formation lies in the formation of a haloamine compound, possibly the mixed dihaloamine bromochloramine.

  14. Reactive films for mitigating methyl bromide emissions from fumigated soil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Emissions of methyl bromide (MeBr) from agricultural fumigation can lead to depletion of the stratospheric ozone layer, and so its use is being phased out. However, as MeBr is still widely used under Critical Use Exemptions, strategies are still required to control such emissions. In this work, nove...

  15. 21 CFR 522.275 - N-Butylscopolammonium bromide.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false N-Butylscopolammonium bromide. 522.275 Section 522.275 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS §...

  16. 21 CFR 522.275 - N-Butylscopolammonium bromide.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false N-Butylscopolammonium bromide. 522.275 Section 522.275 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS §...

  17. 21 CFR 522.275 - N-Butylscopolammonium bromide.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false N-Butylscopolammonium bromide. 522.275 Section 522.275 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS §...

  18. 21 CFR 522.275 - N-Butylscopolammonium bromide.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false N-Butylscopolammonium bromide. 522.275 Section 522.275 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS §...

  19. On the existence of ‘L-alanine cadmium bromide'

    NASA Astrophysics Data System (ADS)

    Srinivasan, Bikshandarkoil R.

    2013-12-01

    It is argued that the recently reported nonlinear optical crystal L-alanine cadmium bromide, grown by slow solvent evaporation method at room temperature [P. Ilayabarathi, J. Chandrasekaran, Spectrochim. Acta 96A (2012) 684-689] is the well-known L-alanine crystal. The isolation of L-alanine crystal is explained due to fractional crystallization.

  20. On the existence of 'L-alanine cadmium bromide'.

    PubMed

    Srinivasan, Bikshandarkoil R

    2013-12-01

    It is argued that the recently reported nonlinear optical crystal L-alanine cadmium bromide, grown by slow solvent evaporation method at room temperature [P. Ilayabarathi, J. Chandrasekaran, Spectrochim. Acta 96A (2012) 684-689] is the well-known L-alanine crystal. The isolation of L-alanine crystal is explained due to fractional crystallization.

  1. 75 FR 5582 - Methyl Bromide; Amendments to Terminate Uses

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-03

    ... or on alfalfa hay and cottonseed for these product registrations. These are the last products containing the pesticide methyl bromide registered for use on alfalfa hay and cotton seed in the United... post-harvest alfalfa hay and post-harvest cottonseed uses is prohibited after October 31, 2009,...

  2. Methyl bromide phase out could affect future reforestation efforts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methyl bromide has long been an integral component in producing healthy tree seedlings in forest nurseries of California, Idaho, Montana, Oregon and Washington. The fumigant was supposed to be completely phased out of use in the United States of America by 2005, but many forest nurseries continue to...

  3. Toxicity study of cerium oxide nanoparticles in human neuroblastoma cells.

    PubMed

    Kumari, Monika; Singh, Shailendra Pratap; Chinde, Srinivas; Rahman, Mohammed Fazlur; Mahboob, Mohammed; Grover, Paramjit

    2014-01-01

    The present study consisted of cytotoxic, genotoxic, and oxidative stress responses of human neuroblastoma cell line (IMR32) following exposure to different doses of cerium oxide nanoparticles (CeO2 NPs; nanoceria) and its microparticles (MPs) for 24 hours. Cytotoxicity was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide and lactate dehydrogenase assays whereas genotoxicity was assessed using the cytokinesis-block micronucleus and comet assays. A battery of assays including lipid peroxidation, reactive oxygen species (ROS), hydrogen peroxide, reduced glutathione, nitric oxide, glutathione reductase, glutathione peroxidase, superoxide dismutase, catalase, and glutathione S-transferase were performed to test the hypothesis that ROS was responsible for the toxicity of nanoceria. The results showed that nanosized CeO2 was more toxic than cerium oxide MPs. Hence, further study on safety evaluation of CeO2 NPs on other models is recommended.

  4. Cytotoxicities and genotoxicities of cements based on calcium silicate and of dental formocresol.

    PubMed

    Ko, Hyunjung; Jeong, Youngdan; Kim, Miri

    2017-03-01

    Increasing interest is being paid to the toxicities of dental materials. The purpose of this study was to determine the cytotoxicities and genotoxicities of endodontic compounds to Chinese hamster ovary (CHO-K1) reproductive cells. Cultured CHO-K1 cells were treated with dental formocresol, two types of calcium hydroxide paste, and two types of mineral trioxide aggregate cement for 24h. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay was performed on each culture, and the micronucleus frequency was determined by performing a micronucleus assay. Alkaline comet assay and γ-H2AX immunofluorescence assay were used to detect DNA damage. Out of the five materials tested, only dental formocresol significantly increased DNA damage. The mineral trioxide aggregate cements based on calcium silicate were not found to be potentially genotoxic. The data suggest that dental formocresol should not be recommended for use in vital pulp therapy on young teeth.

  5. Ex vivo uranium decontamination efficiency on wounded skin and in vitro skin toxicity of a calixarene-loaded nanoemulsion.

    PubMed

    Grives, Sophie; Phan, Guillaume; Morat, Guillaume; Suhard, David; Rebiere, Francois; Fattal, Elias

    2015-06-01

    The present work aims at studying the decontamination efficacy of a calixarene-loaded nanoemulsion on two ex vivo wounded skin models mimicking superficial stings or cuts contaminated with uranium, and on a third model using excoriation. The decontaminating formulation was compared with the currently used radio-decontaminating soapy water (Trait rouge®) treatment. Moreover, to assess skin damage potentially induced by the undiluted nanoemulsion, in vitro toxicity studies were conducted on an in vitro reconstructed human epidermis, coupled with three different toxicity tests [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide, lactate dehydrogenase, and interleukin-1-α]. This work demonstrated not only a significant decontamination activity of the calixarene nanoemulsion on wounded skin, ranging from 92% to 94% of the applied uranium solution according to the ex vivo model used, but also the absence of side effects of this promising treatment.

  6. The MTT assay as tool to evaluate and compare excipient toxicity in vitro on respiratory epithelial cells.

    PubMed

    Scherliess, Regina

    2011-06-15

    There are not many excipients already approved in drug products for the use in the respiratory tract. In this study, a rapid in vitro screening procedure to assess and compare acute toxicity of soluble excipient substances on respiratory epithelial cells utilising the Calu-3 cell line is presented. The test substances are either dissolved in HBSS+HEPES buffer or are directly applied to the cellular surface. After 4h incubation, the substances are removed and the cell viability is assessed using an MTT assay. The tested excipients include polysorbate 20 and 80, lactose and povidone 30 as well as glycerol and propylene glycol as examples of excipients already being used in formulations for application in the respiratory tract. These substances are sorted according to their toxic effect and new excipients not yet used in the respiratory tract like HPMC can be classified in this scheme. With this, besides information from systemic toxicity tests, a first valuation of the acute toxic effect of the substance on respiratory epithelial cells is gained. This can aid in the choice of new excipients being necessary for modern respiratory formulations comprising new active compounds as biomolecules or new delivery strategies such as sustained or prolonged delivery.

  7. Combining the rapid MTT formazan exocytosis assay and the MC65 protection assay led to the discovery of carbazole analogs as small molecule inhibitors of Abeta oligomer-induced cytotoxicity.

    PubMed

    Hong, Hyun-Seok; Maezawa, Izumi; Yao, Nianhuan; Xu, Bailing; Diaz-Avalos, Ruben; Rana, Sandeep; Hua, Duy H; Cheng, R Holland; Lam, Kit S; Jin, Lee-Way

    2007-01-26

    The discovery of small molecule inhibitors of cytotoxicity induced by amyloid-beta (Abeta) oligomers, either applied extracellularly or accumulated intraneuronally, is an important goal of drug development for Alzheimer's disease (AD), but has been limited by the lack of efficient screening methods. Here we describe our approach using two cell-based methods. The first method takes advantage of the unique ability of extracellularly applied Abeta oligomers to rapidly induce the exocytosis of formazan formed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). We employed a short protocol to quantify this toxicity, and quickly identified two novel inhibitors, code-named CP2 and A5, from two compound libraries. A second independent screen of the same libraries using our previously published MC65 protection assay, which identifies inhibitors of toxicity related to intracellular Abeta oligomers, also selected the same two leads, suggesting that both assays select for the same anti-Abeta oligomer properties displayed by these compounds. We further demonstrated that A5 attenuated the progressive aggregation of existing Abeta oligomers, reduced the level of intracellular Abeta oligomers, and prevented the Abeta oligomer-induced death of primary cortical neurons, effects similar to those demonstrated by CP2. Our results suggest that, when combined, the two methods would generate fewer false results and give a high likelihood of identifying leads that show promises in ameliorating Abeta oligomer-induced toxicities within both intraneuronal and extracellular sites. Both assays are simple, suitable for rapid screening of a large number of medicinal libraries, and amenable for automation.

  8. 40 CFR 180.123 - Inorganic bromide residues resulting from fumigation with methyl bromide; tolerances for residues.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... fumigated with the antimicrobial agent and insecticide methyl bromide after harvest (with the exception of... None Pumpkin, postharvest 20.0 None Quince, postharvest 5.0 None Radish, postharvest 30.0 None Rice..., postharvest 20.0 None Turnip, roots, postharvest 30.0 None Walnut, postharvest 200.0 None...

  9. 40 CFR 180.123 - Inorganic bromide residues resulting from fumigation with methyl bromide; tolerances for residues.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... fumigated with the antimicrobial agent and insecticide methyl bromide after harvest (with the exception of... None Pumpkin, postharvest 20.0 None Quince, postharvest 5.0 None Radish, postharvest 30.0 None Rice..., postharvest 20.0 None Turnip, roots, postharvest 30.0 None Walnut, postharvest 200.0 None...

  10. 40 CFR 180.123 - Inorganic bromide residues resulting from fumigation with methyl bromide; tolerances for residues.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... fumigated with the antimicrobial agent and insecticide methyl bromide after harvest (with the exception of... None Pumpkin, postharvest 20.0 None Quince, postharvest 5.0 None Radish, postharvest 30.0 None Rice..., postharvest 20.0 None Turnip, roots, postharvest 30.0 None Walnut, postharvest 200.0 None...

  11. 40 CFR 180.123 - Inorganic bromide residues resulting from fumigation with methyl bromide; tolerances for residues.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... fumigated with the antimicrobial agent and insecticide methyl bromide after harvest (with the exception of... None Pumpkin, postharvest 20.0 None Quince, postharvest 5.0 None Radish, postharvest 30.0 None Rice..., postharvest 20.0 None Turnip, roots, postharvest 30.0 None Walnut, postharvest 200.0 None...

  12. 40 CFR 180.123 - Inorganic bromide residues resulting from fumigation with methyl bromide; tolerances for residues.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... fumigated with the antimicrobial agent and insecticide methyl bromide after harvest (with the exception of..., postharvest 5.0 Radish, postharvest 30.0 Rice, grain, postharvest 50.0 Rutabaga, roots, postharvest 30.0..., roots, postharvest 30.0 Walnut, postharvest 200.0 Watermelon, postharvest 20.0 Wheat 50.0 (2)...

  13. Heat capacities of the water + lithium bromide + ethanolamine and water + lithium bromide + 1,3-propanediol systems

    SciTech Connect

    Kim, J.S.; Park, Y.; Lee, H.; Yu, S.I.

    1997-03-01

    Heat capacities of the water + lithium bromide + ethanolamine (LiBr/H{sub 2}N(CH{sub 2}){sub 2}OH mass ratio = 3.5) and water + lithium bromide + 1,3-propanediol (LiBr/HO(CH{sub 2}){sub 3}OH mass ratio = 3.5) systems were measured by using an isoperibol solution calorimeter at four temperatures (283.15, 298.15, 313.15, and 333.15 K) and absorbent (LiBr + H{sub 2}N(CH{sub 2}){sub 2}OH and LiBr + HO(CH{sub 2}){sub 3}OH) concentration ranges of (29.2 to 70.7)% and (30.7 to 68.3)%, respectively. The measured values were fitted with a simple equation by a least-squares method and the average absolute deviations between experimental and calculated values were 0.21% for the water + lithium bromide + ethanolamine system and 0.15% for the water + lithium bromide + 1,3-propanediol system, respectively.

  14. Mtt1 is a Upf1-like helicase that interacts with the translation termination factors and whose overexpression can modulate termination efficiency.

    PubMed Central

    Czaplinski, K; Majlesi, N; Banerjee, T; Peltz, S W

    2000-01-01

    Translation termination is the final step that completes the synthesis of a polypeptide. Premature translation termination by introduction of a nonsense mutation leads to the synthesis of a truncated protein. We report the identification and characterization of the product of the MTT1 gene, a helicase belonging to the Upfl-like family of helicases that is involved in modulating translation termination. MTT1 is homologous to UPF1, a factor previously shown to function in both mRNA turnover and translation termination. Overexpression of MTT1 induced a nonsense suppression phenotype in a wild-type yeast strain. Nonsense suppression is apparently not due to induction of [PSI+], even though cooverexpression of HSP104 alleviated the nonsense suppression phenotype observed in cells overexpressing MTT1, suggesting a more direct role of Hsp104p in the translation termination process. The MTT1 gene product was shown to interact with translation termination factors and is localized to polysomes. Taken together, these results indicate that at least two members of a family of RNA helicases modulate translation termination efficiency in cells. PMID:10836794

  15. In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride.

    PubMed

    Fotakis, George; Timbrell, John A

    2006-01-05

    The aim of this study was to compare four in vitro cytotoxicity assays and determine their ability to detect early cytotoxic events. Two hepatoma cell lines, namely HTC and HepG2 cells, were exposed to cadmium chloride (0-300 microM) for 3, 5 and 8 h. Following exposure to the toxic metal cytotoxicity was determined with the lactate dehydrogenase leakage assay (LDH), a protein assay, the neutral red assay and the methyl tetrazolium (MTT) assay. In HTC cells no toxicity was observed for any incubation period when the LDH leakage, the MTT and the protein assay were employed whereas the neutral red assay revealed early cytotoxicity starting after incubation of HTC cells with CdCl(2) for 3 h. In the case of HepG2 cells the MTT assay reveals cytotoxicity due to CdCl(2) exposure after 3 h whereas no such effect is seen with the other three assays. Following 5 h exposure of HepG2 cells to CdCl(2), toxicity is observed with the MTT assay at lower concentrations compared to the ones required for detection of toxicity with the LDH leakage and the neutral red assay. In conclusion different sensitivity was observed for each assay with the neutral red and the MTT assay being the most sensitive in detecting cytotoxic events compared to the LDH leakage and the protein assay.

  16. 40 CFR 721.4090 - Ethanaminium, N-[bis(diethylamino)-methylene]-N-ethyl-, bromide.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Ethanaminium, N- -N-ethyl-, bromide... Substances § 721.4090 Ethanaminium, N- -N-ethyl-, bromide. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified as ethanaminium, N- -N-ethyl-, bromide (PMN...

  17. 40 CFR 721.4090 - Ethanaminium, N-[bis(diethylamino)-methylene]-N-ethyl-, bromide.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 32 2012-07-01 2012-07-01 false Ethanaminium, N- -N-ethyl-, bromide... Substances § 721.4090 Ethanaminium, N- -N-ethyl-, bromide. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified as ethanaminium, N- -N-ethyl-, bromide (PMN...

  18. 40 CFR 721.4090 - Ethanaminium, N-[bis(diethylamino)-methylene]-N-ethyl-, bromide.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 31 2014-07-01 2014-07-01 false Ethanaminium, N- -N-ethyl-, bromide... Substances § 721.4090 Ethanaminium, N- -N-ethyl-, bromide. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified as ethanaminium, N- -N-ethyl-, bromide (PMN...

  19. 40 CFR 721.4090 - Ethanaminium, N-[bis(diethylamino)-methylene]-N-ethyl-, bromide.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 32 2013-07-01 2013-07-01 false Ethanaminium, N- -N-ethyl-, bromide... Substances § 721.4090 Ethanaminium, N- -N-ethyl-, bromide. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified as ethanaminium, N- -N-ethyl-, bromide (PMN...

  20. 40 CFR 180.519 - Bromide ion and residual bromine; tolerances for residues.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Bromide ion and residual bromine... Tolerances § 180.519 Bromide ion and residual bromine; tolerances for residues. (a) General. The food additives, bromide ion and residual bromine, may be present in water, potable in accordance with...

  1. 40 CFR 180.519 - Bromide ion and residual bromine; tolerances for residues.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Bromide ion and residual bromine... Tolerances § 180.519 Bromide ion and residual bromine; tolerances for residues. (a) General. The food additives, bromide ion and residual bromine, may be present in water, potable in accordance with...

  2. 40 CFR 180.519 - Bromide ion and residual bromine; tolerances for residues.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Bromide ion and residual bromine... Tolerances § 180.519 Bromide ion and residual bromine; tolerances for residues. (a) General. The food additives, bromide ion and residual bromine, may be present in water, potable in accordance with...

  3. 40 CFR 180.519 - Bromide ion and residual bromine; tolerances for residues.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Bromide ion and residual bromine... Tolerances § 180.519 Bromide ion and residual bromine; tolerances for residues. (a) General. The food additives, bromide ion and residual bromine, may be present in water, potable in accordance with...

  4. 40 CFR 180.519 - Bromide ion and residual bromine; tolerances for residues.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Bromide ion and residual bromine... Tolerances § 180.519 Bromide ion and residual bromine; tolerances for residues. (a) General. The food additives, bromide ion and residual bromine, may be present in water, potable in accordance with...

  5. 40 CFR 721.4090 - Ethanaminium, N-[bis(diethylamino)-methylene]-N-ethyl-, bromide.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Ethanaminium, N- -N-ethyl-, bromide... Substances § 721.4090 Ethanaminium, N- -N-ethyl-, bromide. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified as ethanaminium, N- -N-ethyl-, bromide (PMN...

  6. Uptake of ozone to mixed sodium bromide/ citric acid solutions

    NASA Astrophysics Data System (ADS)

    Lee, Ming-Tao; Steimle, Emilie; Bartels-Rausch, Thorsten; Kato, Shunsuke; Lampimäki, Markus; Brown, Matthew; van Bokhoven, Jeroen; Nolting, Frithjof; Kleibert, Armin; Türler, Andreas; Ammann, Markus

    2013-04-01

    Sea-salt solution - air interfaces play an important role in the chemistry of the marine boundary layer. The reaction of ozone (O3) with bromide is of interest in the context of formation of photolabile halogens (Br2, BrCl) in the marine boundary layer. Recent experiments have suggested that the bromide oxidation rate is related to the surface concentration of bromide [1] and inversely related to the gas phase concentration of O3, an indication for a precursor mediated reaction at the surface [2]. So far, the effect of organics (such as those occurring at the ocean surface or in marine aerosols) on the reaction of O3 with bromide aerosols has not been studied yet. In our study we investigate the uptake kinetics of O3 to a mixed solution of sodium bromide (NaBr) and citric acid (CA), which represents highly oxidized organic compounds present in the environment, with a well-established coated wall flow tube technique, which leads to exposure of the film to O3 allowing the heterogeneous reactions to take place and the loss of O3 being measured. The results indicate that the uptake of O3 to the films with the higher bromide concentrations (0.34M and 4M) is independent of the gas phase concentration and roughly consistent with uptake limited by reaction in the bulk. For the lower bromide concentration (84mM), however, we observe a trend of the uptake coefficient to decrease with increasing O3 concentration, indicating an increasing importance of a surface reaction. In an attempt to constrain the kinetic data, we employed X-ray photoelectron spectroscopy (XPS) to get insight into the surface composition of the aqueous solution - air interface. Previous XPS studies have shown that halide ion concentrations are enhanced at the aqueous solution air interface [3-4], which likely promotes the surface reactions of bromide or iodide with O3. A first XPS study of ternary solutions of KI with butanol indicated the importance of specific interactions of the cation with the alcohol

  7. Study of ferroelectric characteristics of diisopropylammonium bromide films

    NASA Astrophysics Data System (ADS)

    Thirmal, C.; Biswas, P. P.; Shin, Y. J.; Noh, T. W.; Giridharan, N. V.; Venimadhav, A.; Murugavel, P.

    2016-09-01

    Organic molecular ferroelectrics are highly desirable due to their numerous advantages. In the present work, a thick film of diisopropylammonium bromide organic molecular ferroelectric is fabricated on the ITO/glass substrate. The grown film shows preferential orientation along the c-axis with a ferroelectric transition at 419 K. The piezoresponse force microscopic measurements are done in a dual ac resonance tracking mode for its switching characteristics. The amplitude and phase images of the oppositely written domain patterns exhibit a clear contrast with 180° phase difference. The dynamical spectroscopic studies reveal a butterfly loop in amplitude and hysteretic character of the phase which are the expected characteristics features of ferroelectrics. In addition, the macroscopic polarization versus electric field hysteresis gives an additional proof for ferroelectric character of the film with the maximum polarization of 3.5 μC/cm2. Overall, we have successfully fabricated diisopropylammonium bromide organic films and demonstrated its room temperature ferroelectric characteristics.

  8. A novel and robust conditioning lesion induced by ethidium bromide

    PubMed Central

    Hollis, Edmund R; Ishiko, Nao; Tolentino, Kristine; Doherty, Ernest; Rodriguez, Maria J.; Calcutt, Nigel A.; Zou, Yimin

    2015-01-01

    Molecular and cellular mechanisms underlying the peripheral conditioning lesion remain unsolved. We show here that injection of a chemical demyelinating agent, ethidium bromide, into the sciatic nerve induces a similar set of regeneration-associated genes and promotes a 2.7-fold greater extent of sensory axon regeneration in the spinal cord than sciatic nerve crush. We found that more severe peripheral demyelination correlates with more severe functional and electrophysiological deficits, but more robust central regeneration. Ethidium bromide injection does not activate macrophages at the demyelinated sciatic nerve site, as observed after nerve crush, but briefly activates macrophages in the dorsal root ganglion. This study provides a new method for investigating the underlying mechanisms of the conditioning response and suggests that loss of the peripheral myelin may be a major signal to change the intrinsic growth state of adult sensory neurons and promote regeneration. PMID:25541322

  9. Thermal stability of octadecyltrimethylammonium bromide modified montmorillonite organoclay.

    PubMed

    Xi, Yunfei; Zhou, Qin; Frost, Ray L; He, Hongping

    2007-07-15

    Organoclays are significant for providing a mechanism for the adsorption of organic molecules from potable water. As such their thermal stability is important. A combination of thermogravimetric analysis and infrared emission spectroscopy was used to determine this stability. Infrared emission spectroscopy (IES) was used to investigate the changes in the structure and surface characteristics of water and surfactant molecules in montmorillonite, octadecyltrimethylammonium bromide and organoclays prepared with the surfactant octadecyltrimethylammonium bromide with different surfactant loadings. These spectra collected at different temperatures give support to the results obtained from the thermal analysis and also provide additional evidence for the dehydration which is difficult to obtain by normal thermoanalytical techniques. The spectra provide information on the conformation of the surfactant molecules in the clay layers and the thermal decomposition of the organoclays. Infrared emission spectroscopy proved to be a useful tool for the study of the thermal stability of the organoclays.

  10. Aluminium Electroplating on Steel from a Fused Bromide Electrolyte

    SciTech Connect

    Prabhat Tripathy; Laura Wurth; Eric Dufek; Toni Y. Gutknecht; Natalie Gese; Paula Hahn; Steven Frank; Guy Fredrickson; J Stephen Herring

    2014-08-01

    A quaternary bromide bath (LiBr-KBr-CsBr-AlBr3) was used to electro-coat aluminium on steel substrates. The electrolyte was prepared by the addition of AlBr3 into the eutectic LiBr-KBr-CsBr melt. A smooth, thick, adherent and shiny aluminium coating could be obtained with 80 wt.% AlBr3 in the ternary melt. The SEM photographs of the coated surfaces suggest the formation of thick and dense coatings with good aluminium coverage. Both salt immersion and open circuit potential measurement suggest that the coatings did display good corrosion-resistance behavior. Annealing of the coated surfaces, prior to corrosion tests, suggested the robustness of the metallic aluminium coating in preventing the corrosion of the steel surfaces. Studies also indicated that the quaternary bromide plating bath can potentially provide a better aluminium coating on both ferrous and non-ferrous metals, including complex surfaces/geometries.

  11. Aluminum electroplating on steel from a fused bromide electrolyte

    SciTech Connect

    Prabhat K. Tripathy; Laura A. Wurth; Eric J. Dufek; Toni Y. Gutknecht; Natalie J. Gese; Paula Hahn; Steven M. Frank; Guy L. Frederickson; J. Stephen Herring

    2014-08-01

    A quaternary bromide bath (LiBr–KBr–CsBr–AlBr3) was used to electro-coat aluminum on steel substrates. The electrolytewas prepared by the addition of AlBr3 into the eutectic LiBr–KBr–CsBr melt. A smooth, thick, adherent and shiny aluminum coating could be obtained with 80 wt.% AlBr3 in the ternary melt. The SEM photographs of the coated surfaces suggest the formation of thick and dense coatings with good aluminum coverage. Both salt immersion and open circuit potential measurement suggested that the coatings did display a good corrosionresistance behavior. Annealing of the coated surfaces, prior to corrosion tests, suggested the robustness of the metallic aluminum coating in preventing the corrosion of the steel surfaces. Studies also indicated that the quaternary bromide plating bath can potentially provide a better aluminumcoating on both ferrous and non-ferrous metals, including complex surfaces/geometries.

  12. [The compatibility between packing material and ipratropium bromide aerosol].

    PubMed

    Yue, Zhi-hua; Shen, Dian-dian; Hu, Chang-qin

    2010-08-01

    With the establishment of HPLC and LC-MS methods to determine the related substances and the content of active pharmaceutical ingredient (API) in ipratropium bromide aerosol products, several packing material-related impurities were identified, including antioxygen BHT and antioxygen 2246. Results showed that these leachable additives from the packing materials may present at a relative high level in the drug solution, and the low content of API in the drug products is usually due to the adsorption of the packing material as well as the leaking of contents. The current available assay methods for the control of ipratropium bromide aerosol products are often lack of specificity and unable to assure the drug quality effectively. To meet the increasing attention on the regulations of drug packing materials, our research would be a pilot study, indicating that the inappropriate packing materials could cause the migration and adsorption of the active ingredients, and the importance to have compatibility studies between packing materials and drugs.

  13. Effect of bromide and nitrite on the degradation of monochloramine

    SciTech Connect

    Valentine, R.L.; Selleck, R.E.

    1981-10-01

    The results indicate that relatively small concentrations of nitrite can greatly accelerate the degradation of monochloramine in the presence of bromide. It does not appear that nitrite is being significantly consumed in a 1:1 stoichiometric oxidation by monochloramine. If the effect of nitrite is catalytic then these results suggest that the presence of nitrite may also accelerate other oxidation-reduction reactions. For example, nitrite may play an important role in oxidant decay in partially nitrified sewage effluents where both monochloramine and nitrite may be present. If not a complex oxidation-reduction possibly involving bromide as a catalyst is indicated. The results also suggest that the presence of other potentially oxidizable species may affect oxidant decay in a manner not attributable to a simple parallel oxidation.

  14. A Lithium Bromide Absorption Chiller with Cold Storage

    DTIC Science & Technology

    2011-01-15

    TO R A G E A LITHIUM BROMIDE ABSORPTION CHILLER WITH COLD STORAGE William Gerstler, et al, General Electric Global Research UNCLASSIFIED UNLIMITED...Research ABSTRACT A LiBr-based absorption chiller can use waste heat or solar energy to produce useful space cooling for small buildings...However, operating this absorption chiller at high ambient tem- peratures may result in performance degradation, crystallization in the absorber, and

  15. Conservative tracer bromide inhibits pesticide mineralisation in soil.

    PubMed

    Bech, Tina B; Rosenbom, Annette E; Sørensen, Sebastian R; Jacobsen, Carsten S

    2017-03-01

    Bromide is a conservative tracer that is often applied with non-conservative solutes such as pesticides to estimate their retardation in the soil. It has been applied in concentrations of up to 250 g Br L(-1), levels at which the growth of single-celled organisms can be inhibited. Bromide applications may therefore affect the biodegradation of non-conservative solutes in soil. The present study investigated the effect of potassium bromide (KBr) on the mineralisation of three pesticides - glyphosate, MCPA and metribuzin - in four agricultural A-horizon soils. KBr was added to soil microcosms at concentrations of 0, 0.5, 2.5 and 5 g Br(-) L(-1) in the soil solution. The study concluded that KBr had a negative effect on pesticide mineralisation. The inhibitory effect varied depending on the KBr concentration, the type of pesticide and the type of soil. Furthermore, 16 S amplicon sequencing revealed that the KBr treatment generally reduced the abundance of bacteroidetes and proteobacteria on both an RNA and DNA level. Therefore, in order to reduce the effect of KBr on the soil bacterial community and consequently also on xenobiotic degradation, it is recommended that KBr be applied in a concentration that does not exceed 0.5 g Br(-) L(-1) in the soil water.

  16. The Thz Absorption of Methyl Bromide (CH_3BR)

    NASA Astrophysics Data System (ADS)

    Ramos, Marlon; Drouin, Brian J.

    2011-06-01

    The possibility of monitoring Methyl Bromide is of interest for both environmental and health concerns. It has an ozone depletion potential of 0.2% and falls under regulations of the Clean Air Act. Neurological effects from long term exposure may result from its major use as a pesticide. Recent improvements in microwave limb sounding at mm & submm wavelengths have resulted in retrievals of Methyl Chloride from atmospheric spectra. It is conceivable that Methyl Bromide would also be measurable by this technique. In an effort to extend and improve the previous work, the THz spectrum of Methyl Bromide has been measured at JPL. We used an isotopically enriched 13CH_3Br (90%) sample and recorded spectra from 750 - 1200 GHz. Our assignment covers the CH_379Br, CH_381Br, 13CH_379Br and 13CH_381Br isotopologues with J< 66 and K< 17 for the ground vibrational state. We plan to assign vibrational satellites and investigate possible perturbations near K =12 in the ground state.

  17. Quality evaluation of Hypericum ascyron extract by two-dimensional high-performance liquid chromatography coupled with the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.

    PubMed

    Li, Xiu-Mei; Luo, Xue-Gang; Zhang, Chao-Zheng; Wang, Nan; Zhang, Tong-Cun

    2015-02-01

    In this paper, a heart-cutting two-dimensional high-performance liquid chromatography coupled with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was established for controlling the quality of different batches of Hypericum ascyron extract for the first time. In comparison with the common one-dimensional fingerprint, the second-dimensional fingerprint compiled additional spectral data and was hence more informative. The quality of H. ascyron extract was further evaluated by similarity measures and the same results were achieved, the correlation coefficients of the similarity of ten batches of H. ascyron extract were >0.99. Furthermore, we also evaluated the quality of the ten batches of H. ascyron extract by antibacterial activity. The result demonstrated that the quality of the ten batches of H. ascyron extract was not significantly different by MTT. Finally, we demonstrated that the second-dimensional fingerprint coupled with the MTT method was a more powerful tool to characterize the quality of samples of batch to batch. Therefore the proposed method could be used to comprehensively conduct the quality control of traditional Chinese medicines.

  18. Tiotropium Bromide in Chronic Obstructive Pulmonary Disease and Bronchial Asthma.

    PubMed

    Alvarado-Gonzalez, Alcibey; Arce, Isabel

    2015-11-01

    Inhaled bronchodilators are the mainstay of pharmacological treatment for stable chronic obstructive pulmonary disease (COPD), including β2-agonists and muscarinic antagonists. Tiotropium bromide, a long-acting antimuscarinic bronchodilator (LAMA), is a treatment choice for moderate-to-severe COPD; its efficacy and safety have been demonstrated in recent trials. Studies also point to a beneficial role of tiotropium in the treatment of difficult-to-control asthma and a potential function in the asthma-COPD overlap syndrome (ACOS). Combination of different bronchodilator molecules and addition of inhaled corticosteroids are viable therapeutic alternatives. A condensation of the latest trials and the rationale behind these therapies will be presented in this article.

  19. Induced Adsorption of Chloride and Bromide by Submonolayer Amounts of Copper Underpotentially Deposited on Pt(111)

    DTIC Science & Technology

    1994-03-14

    S GD Induced Adsorption Of Chloride And Bromide By Submonolayer Amounts Of Copper Underpotentially Deposited On Pt(111). R. G6mez, J.M. Feliu, and...The underpotential deposition of submonolayer amounts of copper induces an enhanced adsorption of chloride and bromide on Pt(111) and is reflected in...nhinoItO _. •- Induced Adsorption of Chloride and Bromide by Submonolayer Amounts of Copper Underpotentially Deposited on Pt(1 11). R. G6mez1 , J.M

  20. Mixed Iodide-Bromide Methylammonium Lead Perovskite-based Diodes for Light Emission and Photovoltaics.

    PubMed

    Gil-Escrig, Lidón; Miquel-Sempere, Araceli; Sessolo, Michele; Bolink, Henk J

    2015-09-17

    Vacuum deposition techniques are used to prepare mixed iodide-bromide methylammonium lead perovskite diodes via an intermediate double layer of the pure iodide and bromide perovskites. The diodes lead to bright electroluminescence, whose emission spectra maxima shift from the infrared toward the visible with increasing bromide content. When illuminated with AM1.5 simulated sunlight the devices function as efficient solar cells with power conversion efficiencies as high as 12.9%.

  1. Purified high molecular weight synthetic Aβ(1-42) and biological Aβ oligomers are equipotent in rapidly inducing MTT formazan exocytosis.

    PubMed

    Weidner, Adam M; Housley, Molly; Murphy, M Paul; Levine, Harry

    2011-06-15

    Synthetic soluble Aβ oligomers are often used as a surrogate for biologic material in a number of model systems. We compared the activity of Aβ oligomers (synthetic and cell culture media derived) on the human SH-SY5Y neuroblastoma and C2C12 mouse myoblast cell lines in a novel, modified MTT assay. Separating oligomers from monomeric peptide by size exclusion chromatography produced effects at peptide concentrations approaching physiologic levels (10-100 nM). Purified oligomers, but not monomers or fibrils, elicited an increase of a detergent-insoluble form of MTT formazan within 2h as opposed to a control toxin (H(2)O(2)). This effect was comparable for biological and synthetic peptide in both cell types. Monomeric Aβ attenuated the effect of soluble oligomers. This study suggests that the activities of biological and synthetic oligomers are indistinguishable during early stages of Aβ oligomer-cell interaction.

  2. Comparison of Heat and Bromide as Tracers of Stream Exchanges With Shallow Ground Water

    NASA Astrophysics Data System (ADS)

    Constantz, J.; Cox, M. H.; Su, G. W.

    2002-12-01

    Heat and bromide were compared as tracers for examining stream/groundwater exchanges along the middle reaches of the Santa Clara River, CA, during a 10-hour surface-water sodium bromide injection. Three cross-sections comprised of 6 shallow (1 m) piezometers were installed at the upper, middle, and lower sections of a 17 km long study reach to monitor temperatures and bromide concentrations in the shallow ground water beneath the stream. The heat and ground-water transport simulation model, VS2DH, and a closely related solute and ground-water transport simulation model, VS2DT, were matched up for comparison of simulated and observed temperatures and bromide concentrations in the streambed. Simulated sediment temperature were fitted to observed temperature results to yield apparent streambed hydraulic conductivities in each cross-section. Saturated hydraulic conductivities ranged from 1.39 x 10-5 m/s in the upper reach to 5.56 x 10-4 m/s in the lower reach. The temperature-based hydraulic conductivities were inserted into VS2DT to predict sediment bromide concentrations during the sodium bromide injection. The predicted bromide concentration curves in the sediments yielded an excellent match to the observed bromide concentrations, without adjustment of any model parameters. This indicates that for the spatial and temporal scales examined on the Santa Clara River, the use of heat and bromide as tracers provide comparable information with respect to apparent hydraulic conductivities and water fluxes in near-stream environments.

  3. Transient neuromyopathy after bromide intoxication in a dog with idiopathic epilepsy

    PubMed Central

    2012-01-01

    A seven-year old Australian Shepherd, suffering from idiopathic epilepsy under treatment with phenobarbitone and potassium bromide, was presented with generalised lower motor neuron signs. Electrophysiology and muscle-nerve biopsies revealed a neuromyopathy. The serum bromide concentration was increased more than two-fold above the upper reference value. Clinical signs disappeared after applying diuretics and reducing the potassium bromide dose rate. This is the first case report describing electrophysiological and histopathological findings associated with bromide induced lower motor neuron dysfunction in a dog. PMID:23216950

  4. Solubilities of bromide salts of aluminum, cobalt, lead, manganese, potassium, and sodium when sparged with hydrogen bromide

    SciTech Connect

    Noble, E.G.; Shanks, D.E.

    1988-01-01

    The effects of HBr concentration and temperature on the solubility and hydration state of AlBr/sub 3/, CoBr/sub 2/, PbBr/sub 2/, MnBr/sub 2/, KBr, and NaBr were investigated by the Bureau of Mines. Saturated aqueous solutions of the Al, Pb, K, and Na bromides were sparged with HBr gas at 20/sup 0/, 40/sup 0/, and 60/sup 0/C, and saturated solutions of Co and Mn bromides were sparged at 20/sup 0/C. Increased HBr concentration caused decreased salt solubility because of the common ion effect for all the investigated bromides except PbBr/sub 2/. Aqueous solubilities at 20/sup 0/C were, in percent, 51.7 for AlBr/sub 3/, 53.2 for CoBr/sub 2/, 1.1 for PbBr/sub 2/, 59.6 for MnBr/sub 2/, 39.5 for KBr, and 47.4 for Na Br. At 60/sup 0/C,the solubilities were, in percent, 53.3 for AlBr/sub 3/, 2.2 for PbBr/sub 2/, 46.0 for KBr, and 53.8 for NaBr. Solubilities in solutions sparged to HBr saturation at 20/sup 0/C were, in percent, 0.5 for AlBr/sub 3/, 42.0 for CoBr/sub 2/, 32.2 for Br/sub 2/, 21.0 for MnBr/sub 2/, 0.6 for KBr, and 0.3 for NaBr. At 60/sup 0/C, the solubilities at HBr saturation were, in percent, 1.8 for AlBr/sub 3/, 41.4 for PbBr/sub 2/, 1.3 for KBr, and 0.4 for NaBr. CoBr/sub 2/ precipitated out to a minimum solubility of 27.2 pct at 28.7-pct-HBr concentration. Further increases in HBr concentration increased CoBr/sub 2/ solubility because of the formation of bromide complexes. PbBr/sub 2/ increased in solubility as HBr concentration increased.

  5. The in vitro and in vivo profile of aclidinium bromide in comparison with glycopyrronium bromide.

    PubMed

    Gavaldà, Amadeu; Ramos, Israel; Carcasona, Carla; Calama, Elena; Otal, Raquel; Montero, José Luis; Sentellas, Sonia; Aparici, Monica; Vilella, Dolors; Alberti, Joan; Beleta, Jorge; Miralpeix, Montserrat

    2014-08-01

    This study characterised the in vitro and in vivo profiles of two novel long-acting muscarinic antagonists, aclidinium bromide and glycopyrronium bromide, using tiotropium bromide and ipratropium bromide as comparators. All four antagonists had high affinity for the five muscarinic receptor sub-types (M1-M5); aclidinium had comparable affinity to tiotropium but higher affinity than glycopyrronium and ipratropium for all receptors. Glycopyrronium dissociated faster from recombinant M3 receptors than aclidinium and tiotropium but more slowly than ipratropium; all four compounds dissociated more rapidly from M2 receptors than from M3 receptors. In vitro, aclidinium, glycopyrronium and tiotropium had a long duration of action at native M3 receptors (>8 h versus 42 min for ipratropium). In vivo, all compounds were equi-potent at reversing acetylcholine-induced bronchoconstriction. Aclidinium, glycopyrronium and ipratropium had a faster onset of bronchodilator action than tiotropium. Aclidinium had a longer duration of action than glycopyronnium (time to 50% recovery of effect [t½ offset] = 29 h and 13 h, respectively); these compare with a t½ offset of 64 h and 8 h for tiotropium and ipratropium, respectively. Aclidinium was less potent than glycopyrronium and tiotropium at inhibiting salivation in conscious rats (dose required to produce half-maximal effect [ED50] = 38, 0.74 and 0.88 μg/kg, respectively) and was more rapidly hydrolysed in rat, guinea pig and human plasma compared with glycopyrronium or tiotropium. These results indicate that while aclidinium and glycopyrronium are both potent antagonists at muscarinic receptors with similar kinetic selectivity for M3 receptors versus M2, aclidinium has a longer dissociation half-life at M3 receptors and a longer duration of bronchodilator action in vivo than glycopyrronium. The rapid plasma hydrolysis of aclidinium, coupled to its kinetic selectivity, may confer a reduced propensity for systemic

  6. A fast Resazurin-based live viability assay is equivalent to the MTT-test in the KeratinoSens assay.

    PubMed

    Emter, Roger; Natsch, Andreas

    2015-06-01

    The KeratinoSens™ assay was the first cell-based in vitro test in the skin sensitisation adverse outcome pathway to be endorsed by an ECVAM statement. It includes a cell viability assessment, which serves two purposes: It forms part of the prediction model to exclude false-positive irritants and cytotoxicity provides some information on sensitizer potency of chemicals, which can feed into a multivariate potency model. In the KeratinoSens™ protocol, Nrf2-dependent luciferase induction and the MTT-viability assay are performed in parallel plates. Resazurin-based viability assays do not require cell lysis and are compatible with luciferase measurements in the same cells. Here, we performed detailed comparison of the tetrazolium-based MTT assay and the PrestoBlue® assay on 35 reference chemicals tested in the full KeratinoSens™ protocol. Log-transformed IC50 and IC30 values measured with both methods correlate with an R(2) of 0.97 and 0.95. A single chemical showed divergent results and analysis by four different viability assays indicated the PrestoBlue® read-out to be correct. The new more rapid and resource efficient approach has clear advantages: Dose-response curves show lower variability and the two endpoints are measured on the same cells. This approach is a valid addition to or replacement of the MTT-readout in the KeratinoSens™ assay and it is recommended as a general tool for luciferase-based reporter assays.

  7. Predictive value of the surface-enhanced resonance Raman scattering-based MTT assay: a rapid and ultrasensitive method for cell viability in situ.

    PubMed

    Mao, Zhu; Liu, Zhuo; Chen, Lei; Yang, Jin; Zhao, Bing; Jung, Young Mee; Wang, Xu; Zhao, Chun

    2013-08-06

    SERRS (surface-enhanced resonance Raman scattering) has been used to develop and optimize a novel and quantitative MTT assay for living cell viability. This highly sensitive method derives from two factors for formazan signal enhancing: the addition of Au nanoparticles and the resonance effect by 632.8 nm of excitation. The results show that the background elements, such as excessive MTT residues, serum, and the drug, did not interfere with the detection of formazan. Moreover, the detection limit of formazan is as low as 1 ng/mL. With the use of this method to quantify metabolically viable cells, dose-response curves of treated and untreated cells with the drug were constructed on the human lung cancer cell A549. The results also show that the Raman signal generated is dependent on the degree of activation of the cells. In comparison to the traditional method, the main advantages of this method are its rapidity (30 min), high-selectivity, high-precision, and cost-effectiveness (0.1 mg/mL MTT) without time-consuming steps and any modifying or labeling procedure. This work reports on an improved research tool that may help researchers apply this method for in situ cell assays.

  8. Flavonoids, flavonoid metabolites, and phenolic acids inhibit oxidative stress in the neuronal cell line HT-22 monitored by ECIS and MTT assay: a comparative study.

    PubMed

    Kling, Beata; Bücherl, Daniel; Palatzky, Peter; Matysik, Frank-Michael; Decker, Michael; Wegener, Joachim; Heilmann, Jörg

    2014-03-28

    A real-time and label-free in vitro assay based on electric cell-substrate impedance sensing (ECIS) was established, validated, and compared to an end-point MTT assay within an experimental trial addressing the cytoprotective effects of 19 different flavonoids, flavonoid metabolites, and phenolic acids and their methyl esters on the HT-22 neuronal cell line, after induction of oxidative stress with tert-butyl hydroperoxide. Among the flavonoids under study, only those with a catechol unit and an additional 4-keto group provided cytoprotection. The presence of a 2,3-double bond was not a structural prerequisite for a neuroprotective effect. In the case of the phenolics, catechol substitution was the only structural requirement for activity. The flavonoids and other phenolics with a ferulic acid substitution or a single hydroxy group showed no activity. Electrochemical characterization of all compounds via square-wave voltammetry provided a rather specific correlation between cytoprotective activity and redox potential for the active flavonoids, but not for the active phenolics with a low molecular weight. Moreover this study was used to compare label-free ECIS recordings with results of the established MTT assay. Whereas the former provides time-resolved and thus entirely unbiased information on changes of cell morphology that are unequivocally associated with cell death, the latter requires predefined exposure times and a strict causality between metabolic activity and cell death. However, MTT assays are based on standard lab equipment and provide a more economic way to higher throughput.

  9. Bromoform formation in ozonated groundwater containing bromide and humic substances

    SciTech Connect

    Cooper, W.J.; Amy, G.L.; Moore, C.A.; Zika, R.G.

    1986-01-01

    The effect of bromide ion, organic carbon concentration (natural aquatic humic substances), pH, and solar irradiation on the formation of bromoform in ozonated groundwater has been studied. The studies were conducted on four unique samples of groundwater taken from different regions of the Biscayne Aquifer in southern Florida. All other conditions being equal, increases in bromide ion concentrations resulted in increases in CHBr/sub 3/ formation. In three of the four samples, CHBr/sub 3/ formation decreased as the pH level increased from 5 to 9. The fourth sample exhibited an opposite trend whereby the CHBr/sub 3/ concentration increased with increasing pH. Bromoform concentration increased with increased O/sub 3/ concentration over an ozone dosage range of 3.4 to 6.7 mg/L. Ozonated samples placed in sunlight immediately after ozone addition showed a decrease in the formation of CHBr/sub 3/ presumably due to the photodecomposition of HOBr/OBr.

  10. A drinking water utility's perspective on bromide, bromate, and ozonation.

    PubMed

    Bonacquisti, Thomas P

    2006-04-17

    Application of ozone in drinking water treatment plants in the US is growing because of ozone's multiple benefits. Ozone functions as a powerful oxidizing agent and disinfecting agent, it improves finished water quality by reducing turbidity, it reduces the formation of many halogenated disinfection by-products, and it is capable of treating chlorine resistant organisms like cryptosporidia. However, when bromide ion is present, e.g. from the geology, runoff, or sea water intrusion, ozone will convert some of the bromide to bromate depending upon the treatment reaction conditions. Bromate can also be introduced into drinking water as a contaminant in the chlorine used for disinfection. The current maximum contaminant level (MCL) in the USA is 0.010 mg/L, and the maximum contaminant level goal (MCLG) is zero, because of the possibility that bromate may function as a genotoxic carcinogen. The level of the MCL, especially if it is lowered, will significantly impact the ability of many water suppliers to utilize ozone in their water treatment processes and also raise the costs of those applications.

  11. Octahedral Rotation Preferences in Perovskite Iodides and Bromides.

    PubMed

    Young, Joshua; Rondinelli, James M

    2016-03-03

    Phase transitions in ABX3 perovskites are often accompanied by rigid rotations of the corner-connected BX6 octahedral network. Although the mechanisms for the preferred rotation patterns of perovskite oxides are fairly well recognized, the same cannot be said of halide variants (i.e., X = Cl, Br, or I), several of which undergo an unusual displacive transition to a tetragonal phase exhibiting in-phase rotations about one axis (a(0)a(0)c(+) in Glazer notation). To discern the chemical factors stabilizing this unique phase, we investigated a series of 12 perovskite bromides and iodides using density functional theory calculations and compared them with similar oxides. We find that in-phase tilting provides a better arrangement of the larger bromide and iodide anions, which minimizes the electrostatic interactions, improves the bond valence of the A-site cations, and enhances the covalency between the A-site metal and Br(-) or I(-) ions. The opposite effect is present in the oxides, with out-of-phase tilting maximizing these factors.

  12. Deswelling kinetics of polyacrylate gels in solutions of cetyltrimethylammonium bromide.

    PubMed

    Nilsson, Peter; Hansson, Per

    2007-08-23

    The deswelling kinetics of single sodium polyacrylate gel beads (radius 40-160 microm) in aqueous solutions of cetyltrimethylammonium bromide under conditions of forced convection are investigated using micromanipulator assisted light microscopy. The purpose of the study is to further evaluate a previously published model (J. Phys. Chem. B 2003, 107, 9203) using a higher homolog surfactant. For gels with expected fast deswelling (small gel size/low surfactant concentration) and/or in low electrolyte concentration, the model is found to correctly predict the deswelling characteristics of the gel beads. However, for some gels with expected slow deswelling, especially in high electrolyte concentration (10 mM NaBr), the model widely underestimates the required deswelling time. The reason for this is argued to be the longer time frame and high bromide concentration allowing the formation of a denser, more ordered structure in the surface phase, which resists the deformation and reorganization of material necessary for deswelling. Unexpectedly long lag times before the start of deswelling are also found for gels in low surfactant concentration, indicating that a relatively high surfactant concentration in the gel, greatly exceeding the critical aggregation concentration, is needed to start formation of a collapsed surface phase. This critical surfactant concentration is found to be dependent on initial gel radius, as small gels require a relatively higher concentration to initiate collapse.

  13. Validation of an automated fluorescein method for determining bromide in water

    USGS Publications Warehouse

    Fishman, M. J.; Schroder, L.J.; Friedman, L.C.

    1985-01-01

    Surface, atmospheric precipitation and deionized water samples were spiked with ??g l-1 concentrations of bromide, and the solutions stored in polyethylene and polytetrafluoroethylene bottles. Bromide was determined periodically for 30 days. Automated fluorescein and ion chromatography methods were used to determine bromide in these prepared samples. Analysis of the data by the paired t-test indicates that the two methods are not significantly different at a probability of 95% for samples containing from 0.015 to 0.5 mg l-1 of bromide. The correlation coefficient for the same sets of paired data is 0.9987. Recovery data, except for the surface water samples to which 0.005 mg l-1 of bromide was added, range from 89 to 112%. There appears to be no loss of bromide from solution in either type of container.Surface, atmospheric precipitation and deionized water samples were spiked with mu g l** minus **1 concentrations of bromide, and the solutions stored in polyethylene and polytetrafluoroethylene bottles. Bromide was determined periodically for 30 days. Automated fluorescein and ion chromatography methods were used to determine bromide in these prepared samples. Analysis of the data by the paired t-test indicates that the two methods are not significantly different at a probability of 95% for samples containing from 0. 015 to 0. 5 mg l** minus **1 of bromide. The correlation coefficient for the same sets of paired data is 0. 9987. Recovery data, except for the surface water samples to which 0. 005 mg l** minus **1 of bromide was added, range from 89 to 112%. Refs.

  14. Optical and Rheological Properties of a Semi-Diluted Equimolar Solution of Cetyltrimethylammonium Bromide and Potassium Bromide

    NASA Astrophysics Data System (ADS)

    Decruppe, J. P.; Cappelaere, E.; Cressely, R.

    1997-02-01

    We report in this paper the results of rheological and optical experiments performed on a semi-diluted viscoelastic solution of cetyltrimethylammonium bromide (CTAB) in water with sodium bromide at a concentration of 0.3 M l^{-1} in surfactant and salt. This concentration in surfactant is much smaller than the concentration which gives a nematic phase at rest. The solution behaves like a Maxwell fluid with a single relaxation time. When submitted to a shear flow, this solution shows a behaviour typical of a system undergoing a phase transition. When observed between crossed polarizer and analyzer, the gap of the Couette cell appears divided in two bands the optical properties of which are different; in rheology, the curves σ(dot{γ}) are also characteristic of these systems where the shear stress presents a plateau extending between two critical values, dot{γ}_{1c}, and dot{γ}_{2c} of the shear rate. We finally compare the results with previous experiments performed on a concentrated solution of CTAB containing no salt.

  15. Budget of Methyl Bromide in the Atmosphere: Isotopic Constraints

    NASA Astrophysics Data System (ADS)

    Bill, M.; Miller, L. G.; Rhew, R. C.; Goldstein, A. H.

    2001-12-01

    Bromine radicals contribute significantly to stratospheric ozone loss through coupled reactions with ClO, HO2, and NO2 radicals. Bromine is approximately 40-100 times more effective, atom for atom, at destroying ozone than chlorine. For instance, Br coupled reactions are responsible for 30 to 50% of the total ozone loss in the polar vortex. The largest source of bromine to the stratosphere is methyl bromide (CH3Br). CH3Br has a concentration in the troposphere of about 10 pptv, a total estimated lifetime of 0.6 to 0.9 years, and has the highest concentration of any long lived organobromine. Unlike chlorofluorocarbons, which are produced entirely by humans, methyl bromide is produced by both anthropogenic and natural processes. We are developing the use of stable isotopes to constrain the budget of CH3Br through quantification of the source signatures and the isotopic fractionations associated with sinks. The largest natural sources appear to be biological production in oceans ( ~35%), biomass burning ( ~13%), and salt marshes ( ~10%). Thus far, the only natural emissions to be isotopically characterized is from salt marsh plants. Carbon isotopic ratios of CH3Br emitted from the salt marsh have a strong diurnal variation from -65\\permil during daytime when emission rates are highest to -12\\permil at night when emissions are ~13% of the daytime rates. The \\delta13C weighted mean of salt marsh emission is -43\\permil CH3Br. Anthropogenically produced CH3Br is used for fumigation of soils, harvested crops and structures, and represents approximately 30% of the total source flux. The fumigation of harvested crops and structures constitutes approximately one third of the anthropogenic source to the atmosphere and, because the release rate of applied CH3Br approaches 100%, it should have a mean isotopic composition equal to industrially manufactured CH3Br, -54.4\\permil. However during soil fumigation, some of the CH3Br is consumed insitu causing isotopic fractionation

  16. The first barium tin(II) bromide fluoride

    NASA Astrophysics Data System (ADS)

    Dénès, Georges; Merazig, Hocine; Muntasar, Abdualhafeed; Porterfield, Robyn

    2014-04-01

    In an effort to prepare barium tin(II) bromide fluorides for the first time, possibly similar to the chloride fluorides obtained earlier in our laboratory, precipitation reactions were carried out by mixing aqueous solutions of SnF2 and of BaBr2.2H2O. In contrast with the chloride fluoride system, a single powdered phase was obtained throughout the SnF2 - BaBr2 system, with the yield being maximum at X ≈ 0.25, where X is the molar fraction of barium bromide in the reaction mixture. Phase identification with the JCPDS database failed to produce a match, confirming that a new phase had been produced. The exact chemical composition of the new compound has not been obtained yet. Based on the X value for the maximum yield, the Sn/Ba ratio is likely to be 3/1 or 2/1. The Mössbauer spectrum at ambient conditions shows that bonding to tin(II) is covalent, therefore with the tin lone pair being stereoactive. The Mössbauer parameters ( δ = 3.68 mm/s, Δ = 0.99 mm/s) are similar to those of SnBrF and of Sn2BrF5, thereby showing that tin is bonded to both fluorine and bromine. The larger isomer shift and lower quadrupole splitting than in tin(II) fluorides show that the stereoactivity of the tin lone pair is lower than in the fluorides. The Mössbauer parameters fit well the linear correlation of the quadrupole splitting versus the isomer shift" that has been shown to be present in other series of tin(II) compounds. The linear decrease on this correlation shows that the contribution of non-spherical orbitals ( p and d) to the lone pair is a much larger contributor to the quadrupole splitting than lattice distortions. The structure is likely made of Ba2+ cations and tin(II) fluoride bromide polyatomic anions, with covalent bonding withinthe anions.

  17. Solid phase microextraction for active or passive sampling of methyl bromide during fumigations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The high diffusivity and volatility of methyl bromide make it an ideal compound for Solid Phase Micro Extraction (SPME)-based sampling of air prior to gas-chromatographic quantifications. SPME fibers can be used as active methyl bromide samplers, with high capacities and an equilibrium time of 1-2 m...

  18. Synergistic Actions of Pyridostigmine Bromide and Insecticides on Muscle and Vascular Nociceptors

    DTIC Science & Technology

    2014-01-01

    TITLE: Synergistic Actions of Pyridostigmine Bromide and Insecticides on Muscle and Vascular Nociceptors PRINCIPAL INVESTIGATOR: Brian...pyridostigmine bromide (PB) and a variety of insecticides /repellants co-varied with the development of this condition (Research Advisory Committee on Gulf War...might occur between insecticides , repellants and nerve agent prophylactics used during the GW could modify or damage the machinery of protein

  19. Destruction of methyl bromide sorbed to activated carbon by thiosulfate and electrolysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methyl bromide is widely used as a fumigant for post-harvest and quarantine uses at port facilities due to the low treatment times required, but it is vented to the atmosphere after its use. Due to the potential contributions of methyl bromide to stratospheric ozone depletion, technologies for the c...

  20. 75 FR 41177 - Protection of Stratospheric Ozone: Request for Methyl Bromide Critical Use Exemption Applications...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-15

    ... 2013 control period? U.S. consumption of methyl bromide in the U.S. has declined significantly over the... Act, 42 U.S.C. 7671c. Under EPA implementing regulations, methyl bromide production and consumption... Montreal Protocol provides that the Parties may exempt ``the level of production or consumption that...

  1. Classifying the Basic Parameters of Ultraviolet Copper Bromide Laser

    NASA Astrophysics Data System (ADS)

    Gocheva-Ilieva, S. G.; Iliev, I. P.; Temelkov, K. A.; Vuchkov, N. K.; Sabotinov, N. V.

    2009-10-01

    The performance of deep ultraviolet copper bromide lasers is of great importance because of their applications in medicine, microbiology, high-precision processing of new materials, high-resolution laser lithography in microelectronics, high-density optical recording of information, laser-induced fluorescence in plasma and wide-gap semiconductors and more. In this paper we present a statistical study on the classification of 12 basic lasing parameters, by using different agglomerative methods of cluster analysis. The results are based on a big amount of experimental data for UV Cu+ Ne-CuBr laser with wavelengths 248.6 nm, 252.9 nm, 260.0 nm and 270.3 nm, obtained in Georgi Nadjakov Institute of Solid State Physics, Bulgarian Academy of Sciences. The relevant influence of parameters on laser generation is also evaluated. The results are applicable in computer modeling and planning the experiments and further laser development with improved output characteristics.

  2. Accommodation coefficient of HOBr on deliquescent sodium bromide aerosol particles

    NASA Astrophysics Data System (ADS)

    Wachsmuth, M.; Gäggeler, H. W.; von Glasow, R.; Ammann, M.

    2002-06-01

    Uptake of HOBr on sea salt aerosol, sea salt brine or ice is believed to be a key process providing a source of photolabile bromine (Br2) and sustaining ozone depletion cycles in the Arctic troposphere. In the present study, uptake of HOBr on sodium bromide (NaBr) aerosol particles was investigated at an extremely low HOBr concentration of 300 cm-3 using the short-lived radioactive isotopes 83-86Br. Under these conditions, at maximum one HOBr molecule was taken up per particle. The rate of uptake was clearly limited by the mass accommodation coefficient, which was calculated to be 0.6 ± 0.2. This value is a factor of 10 larger than estimates used in earlier models. The atmospheric implications are discussed using the box model "MOCCA'', showing that the increase of the accommodation coefficient of HOBr by a factor of 10 only slightly affects net ozone loss, but significantly increases chlorine release.

  3. Theoretical and experimental studies of silver bromide clusters

    NASA Astrophysics Data System (ADS)

    Zhang, Hongguang

    2000-11-01

    This work consists of three parts: Experimental synthesis of silver bromide clusters via the electroporation of unilamellar dioleoylphosphatidylcholine (DOPC) vesicles, theoretical study of the structure and electronic properties of (AgBr)n clusters, and characterization of silver bromide clusters grown and their interaction with the surface of DOPC vesicles. In the first part, we successfully synthesized AgBr quantum dots via the electroporation of vesicles. For the first time, we observed the entire blue-shift followed by red-shift of the absorption band (274nm --> 5h 269nm --> 6h 273nm) that is associated with the growth of the silver bromide clusters. The turn-around point is at 269 nm. In the second part, we have theoretically investigated the structures and UV absorption spectra of (AgBr)n clusters (n = 1-9) in both the gas phase and in a dielectric medium. The structures of clusters were determined at the B3P86/SB level with full geometry optimization. For clusters with n = 1-6, extensive searches of the potential energy surface yielded only one minimum, while larger clusters displayed two or more minima. UV absorption spectra were calculated by the configuration interaction singlet (CIS) method with a much larger basis set. Our computational results parallel the experimental trends, and show that the turn- around point occurs at the trimer according to HOMO-LUMO gap calculation and at the tetramer according to CIS calculation. The molecular origin of the blue/red shift associated with AgBr cluster growth can be readily explained by examining the orbital interactions which dominate the process and by the structure of the clusters. The detailed molecular orbital energy level correlation diagram for the dimerization, trimerization and tetramerization will be presented. In the third part, TEM characterization of the quantum dots show

  4. Error Evaluation of Methyl Bromide Aerodynamic Flux Measurements

    USGS Publications Warehouse

    Majewski, M.S.

    1997-01-01

    Methyl bromide volatilization fluxes were calculated for a tarped and a nontarped field using 2 and 4 hour sampling periods. These field measurements were averaged in 8, 12, and 24 hour increments to simulate longer sampling periods. The daily flux profiles were progressively smoothed and the cumulative volatility losses increased by 20 to 30% with each longer sampling period. Error associated with the original flux measurements was determined from linear regressions of measured wind speed and air concentration as a function of height, and averaged approximately 50%. The high errors resulted from long application times, which resulted in a nonuniform source strength; and variable tarp permeability, which is influenced by temperature, moisture, and thickness. The increase in cumulative volatilization losses that resulted from longer sampling periods were within the experimental error of the flux determination method.

  5. Reactions of buffers in cyanogen bromide-induced ligations.

    PubMed

    Vogel, Heike; Gerlach, Claudia; Richert, Clemens

    2013-01-01

    Rapid, template-directed ligation reactions between a phosphate-terminated oligonucleotide and an unphosphorylated reaction partner may be induced by cyanogen bromide (BrCN). Frequently, however, the reaction is low yielding, and even a large excess of the condensing agent can fail to induce quantitative conversions. In this study, we used BrCN to induce chemical primer extension reactions. Here, we report that buffers containing hydroxyl groups react with short oligodeoxynucleotides in the presence of BrCN. One stable adduct between HEPBS buffer and cytosine was characterized by mass spectrometry and NMR after HPLC purification, indicating that a side reaction occurred at this nucleobase. Further, a first example of a primer extension reaction between an unmodified oligodeoxynucleotide as primer and dGMP is reported. Together, our results shed light on the potency, as well as the drawbacks of BrCN as a highly reactive condensing reagent for the ligation of unmodified nucleic acids.

  6. Location of Bromide Ions in Tetragonal Lysozyme Crystals

    NASA Technical Reports Server (NTRS)

    Lim, Kap; Nadarajah, Arunan; Forsythe, Elizabeth L.; Pusey, Marc L.

    1998-01-01

    Anions have been shown to play a dominant role in the crystallization of chicken egg white lysozyme from salt solutions. Previous studies employing X-ray crystallography had found one chloride ion binding site in the tetragonal crystal form of the protein and four nitrate ion binding sites in the monoclinic form. In this study the anion positions in the tetragonal form were determined from the difference Fourier map obtained from lysozyme crystal grown in bromide and chloride solutions. Five possible anion binding sites were found in this manner. Some of these sites were in pockets containing basic residues while others were near neutral, but polar, residues. The sole chloride ion binding site found in previous studies was confirmed, while four of these sites corresponded to four binding sites found for nitrate ions in monoclinic crystals. The study suggests that most of the anion binding sites in lysozyme remain unchanged, even when different anions and different crystal forms of lysozyme are employed.

  7. Unveiling Residual Molecular Binding in Triply Charged Hydrogen Bromide

    SciTech Connect

    Penent, F.; Lablanquie, P.; Palaudoux, J.; Gamblin, G.; Carniato, S.; Andric, L.; Hikosaka, Y.; Ito, K.

    2011-03-11

    We present an experimental and theoretical study of triply charged hydrogen bromide ions formed by photoionization of the inner 3d shell of Br. The experimental results, obtained by detecting the 3d photoelectron in coincidence with the two subsequent Auger electrons, are analyzed using calculated potential energy curves of HBr{sup 3+}. The competition between the short-range chemical binding potential and the Coulomb repulsion in the dissociative process is shown. Two different mechanisms are observed for double Auger decay: one, a direct process with simultaneous ejection of two Auger electrons to final HBr{sup 3+} ionic states and the other, a cascade process involving double Auger decay characterized by the autoionization of Br*{sup +} ion subsequent to the HBr{sup 2+} fragmentation.

  8. Effects of pyridostigmine bromide on in-flight aircrew performance.

    PubMed

    Gawron, V J; Schiflett, S G; Miller, J C; Slater, T; Ball, J F

    1990-02-01

    The effects of a chemical defense pretreatment drug, pyridostigmine bromide (PB), on in-flight aircrew performance were assessed using the Total In-Flight Simulator (TIFS) aircraft. TIFS was used to supply appropriate control dynamics, handling characteristics, and cockpit instrumentation for a tactical transport airdrop simulation. Twenty-one C-130 pilots flew two familiarization and four data flights. During two data flights PB was given to both members of the aircrew using the dosage regimen of 30 mg/8 h prescribed by the U.S. Air Force surgeon general. The drug was administered using a double-blind technique. The results indicated that (1) aircrews successfully completed their assigned mission, (2) airdrop inaccuracies and navigation errors in time and distance were not specifically related to PB, (3) performance and crew coordination were not affected by PB, (4) PB and pilot/copilot not discriminate beyond chance between PB and placebo conditions.

  9. Tiotropium Bromide in Chronic Obstructive Pulmonary Disease and Bronchial Asthma

    PubMed Central

    Alvarado-Gonzalez, Alcibey; Arce, Isabel

    2015-01-01

    Inhaled bronchodilators are the mainstay of pharmacological treatment for stable chronic obstructive pulmonary disease (COPD), including β2-agonists and muscarinic antagonists. Tiotropium bromide, a long-acting antimuscarinic bronchodilator (LAMA), is a treatment choice for moderate-to-severe COPD; its efficacy and safety have been demonstrated in recent trials. Studies also point to a beneficial role of tiotropium in the treatment of difficult-to-control asthma and a potential function in the asthma-COPD overlap syndrome (ACOS). Combination of different bronchodilator molecules and addition of inhaled corticosteroids are viable therapeutic alternatives. A condensation of the latest trials and the rationale behind these therapies will be presented in this article. PMID:26491494

  10. N-(2-Bromo-benz-yl)cinchoninium bromide.

    PubMed

    Skórska-Stania, Agnieszka; Jezierska-Zięba, Magdalena; Kąkol, Barbara; Fedoryński, Michał; Oleksyn, Barbara J

    2012-09-01

    The title compound {systematic name: 1-(2-bromo-benz-yl)-5-ethenyl-2-[hy-droxy(quinolin-4-yl)meth-yl]-1-aza-bicyclo-[2.2.2]octan-1-ium bromide}, C(26)H(28)BrN(2)O(+)·Br(-), is a chiral quater-nary ammonium salt of one of the Cinchona alkaloids. The planes of the quinoline and of the bromo-benzyl substituent are inclined to one another by 9.11 (9)°. A weak intra-molecular C-H⋯O hydrogen bond occurs. The crystal structure features strong O-H⋯Br hydrogen bonds and weak C-H⋯Br inter-actions.

  11. N-(2-Bromo­benz­yl)cinchoninium bromide

    PubMed Central

    Skórska-Stania, Agnieszka; Jezierska-Zięba, Magdalena; Kąkol, Barbara; Fedoryński, Michał; Oleksyn, Barbara J.

    2012-01-01

    The title compound {systematic name: 1-(2-bromo­benz­yl)-5-ethenyl-2-[hy­droxy(quinolin-4-yl)meth­yl]-1-aza­bicyclo­[2.2.2]octan-1-ium bromide}, C26H28BrN2O+·Br−, is a chiral quater­nary ammonium salt of one of the Cinchona alkaloids. The planes of the quinoline and of the bromo­benzyl substituent are inclined to one another by 9.11 (9)°. A weak intra­molecular C—H⋯O hydrogen bond occurs. The crystal structure features strong O—H⋯Br hydrogen bonds and weak C—H⋯Br inter­actions. PMID:22969676

  12. Electrochemical and spectroscopic study of octadecyltrimethylammonium bromide/DNA surfoplexes.

    PubMed

    Rodríguez-Pulido, Alberto; Aicart, Emilio; Junquera, Elena

    2009-04-21

    The use of cationic micelles consisting of octadecyltrimethylammonium bromide (C18TAB) to compact calf thymus DNA has been investigated in aqueous buffered solution at 310.15 K by means of conductometry, electrophoretic mobility, and several fluorescence spectroscopy methods. The results indicate that C18TAB micelles, consisting of 44 monomers on average, may compact DNA molecule by an electrostatic interaction that takes place at the cationic spherical micelle surface. The surfoplexes thus formed show a surface density charge that goes from negative to positive values at a Lmic/D mass ratio of around 1.0 (where Lmic and D are the masses of micellized cationic surfactant and DNA), called the isoneutrality ratio (Lmic/D)phi. Values of this characteristic parameter, determined in this work not only from the electrochemical experimental data but also from spectroscopic measurements, are in very good agreement with those ones calculated from molecular parameters and some other properties also obtained in this work. The electrostatic character of the DNA-micelle interaction has been confirmed by analyzing the decrease in fluorescence emission of the fluorophore ethidium bromide, EtBr, initially intercalated between DNA base pairs, as long as the surfoplexes are formed. Fluorescence anisotropy experiments have revealed that micelle packing becomes more rigid in the presence of DNA, but once the surfoplex is formed, the fluidity increases with the Lmic/D mass ratio, attaining its maximum when the isoneutrality ratio is exceeded. This fact, together with the net positive charge of the surfoplexes with the Lmic/D mass ratio over the isoneutrality ratio, makes this regimen of lipid and DNA content the optimum for efficiency in the transfection process.

  13. Comparative mobility of sulfonamides and bromide tracer in three soils

    USGS Publications Warehouse

    Kurwadkar, S.T.; Adams, C.D.; Meyer, M.T.; Kolpin, D.W.

    2011-01-01

    In animal agriculture, sulfonamides are one of the routinely used groups of antimicrobials for therapeutic and sub-therapeutic purposes. It is observed that, the animals when administered the antimicrobials, often do not completely metabolize them; and excrete the partially metabolized forms into the environment. Due to the continued use of antimicrobials and disposal of untreated waste, widespread occurrence of partially metabolized antimicrobials in aquatic and terrestrial environments has been reported in various scientific journals. In this research, the mobility of two sulfonamides - sulfamethazine (SMN), sulfathiazole (STZ) and a conservative bromide tracer was investigated in three soils collected from regions in the United States with large number of concentrated animal-feed operations. Results of a series of column studies indicate that the mobility of these two sulfonamides was dependent on pH, soil charge density, and contact time. At low pH and high charge density, substantial retention of sulfonamides was observed in all three soils investigated, due to the increased fraction of cationic and neutral forms of the sulfonamides. Conversely, enhanced mobility was observed at high pH, where the sulfonamides are predominantly in the anionic form. The results indicate that when both SMN and STZ are predominantly in anionic forms, their mobility approximates the mobility of a conservative bromide tracer. This observation is consistent for the mobility of both SMN and STZ individually, and also in the presence of several other antimicrobials in all three soils investigated. Higher contact time indicates lower mobility due to increased interaction with soil material. ?? 2011.

  14. Nebulised ipratropium bromide and sodium cromoglycate in the first two years of life.

    PubMed Central

    Henry, R L; Hiller, E J; Milner, A D; Hodges, I G; Stokes, G M

    1984-01-01

    In a double blind crossover trial, we compared sodium cromoglycate, ipratropium bromide, and water in 23 asthmatic children less than 2 years old (mean age 11.8 months). Each child received nebulised solutions containing 20 mg of sodium cromoglycate, 250 micrograms of ipratropium bromide, or 2 ml water three times a day for three two month periods. Daily symptom scores did not show significant differences between the treatments but parental preferences indicated that both sodium cromoglycate and ipratropium bromide were superior to placebo. Sodium cromoglycate was prophylactic and was more likely to help the older patients. Ipratropium bromide produced an immediate clinical benefit and the response was not age dependent. We were unable to pick responders from non-responders on the basis of lung function tests performed on a routine outpatient basis. Both ipratropium bromide and sodium cromoglycate help some but not all asthmatic children aged less than 2 years. PMID:6230059

  15. X-Ray absorption spectroscopy investigation of 1-alkyl-3-methylimidazolium bromide salts

    SciTech Connect

    D'Angelo, Paola; Zitolo, Andrea; Migliorati, Valentina; Bodo, Enrico; Caminiti, Ruggero; Aquilanti, Giuliana; Hazemann, Jean Louis; Testemale, Denis; Mancini, Giordano

    2011-08-21

    X-ray absorption spectroscopy (XAS) has been used to unveil the bromide ion local coordination structure in 1-alkyl-3-methylimidazolium bromide [C{sub n}mim]Br ionic liquids (ILs) with different alkyl chains. The XAS spectrum of 1-ethyl-3-methylimidazolium bromide has been found to be different from those of the other members of the series, from the butyl to the decyl derivatives, that have all identical XAS spectra. This result indicates that starting from 1-buthyl-3-methylimidazolium bromide the local molecular arrangement around the bromide anion is the same independently from the length of the alkyl chain, and that the imidazolium head groups in the liquid ILs with long alkyl chains assume locally the same orientation as in the [C{sub 4}mim]Br crystal. With this study we show that the XAS technique is an effective direct tool for unveiling the local structural arrangements around selected atoms in ILs.

  16. Bromide's effect on DBP formation, speciation, and control; Part 1: Ozonation

    SciTech Connect

    Shukairy, H.M.; Summers, R.S. ); Miltner, R.J. . Drinking Water Research Div.)

    1994-06-01

    The effect of variable ozone dosage and bromide concentration on the formation of organic disinfection by-products (DBPs) and bromate were evaluated. Low ozone dosages resulted in oxidation of organic precursors, yielding decreases in the formation potential for total trihalomethanes (THMs), six haloacetic acids (HAAs), and total organic halide (TOX). Increasing the ozone dosage oxidized bromide to bromate, decreasing the bromide for incorporation into DBPs. Bromate concentrations were linearly correlated with ozone residuals. Changes in the bromine incorporation factors n and n[prime] reflected differences in the resulting speciation of THMs and HAAs, respectively. Because TOX measurements based on chloride equivalence may underestimate the halogenated DBP yield for high-bromide waters, a procedure is described whereby bromide and bromate concentrations were used to correct the TOX measurement.

  17. In vitro assessment of sperm from bulls of high and low field fertility.

    PubMed

    Al Naib, A; Hanrahan, J P; Lonergan, P; Fair, S

    2011-07-01

    The aim of this study was to investigate the reasons for differences in field fertility of bulls following insemination with frozen-thawed semen. The study was carried out in two separate parts over two years and comparisons were made between 5 high and 4 low fertility Holstein Friesian bulls as determined by their either 90 day non-return rate (Year 1) or calving rate (Year 2). Two high fertility Limousin bulls were included in Year 1 for comparative purposes. The ability of sperm from each bull to penetrate artificial mucus was assessed (Year 1 = 7 replicates; Year 2 = 5 replicates). Glass capillary tubes (2 per bull per replicate) were filled with artificial mucus and incubated with sperm stained in 1% Hoechst 33342 for 30 min at 37 °C. The number of sperm were subsequently counted at 10 mm intervals along the tube between 40 and 80 mm markers. Sperm mitochondrial activity of each bull was assessed by the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assay (4 replicates in each year). Sperm were incubated with MTT for 1 h at 37 °C following which the absorbance of formazan was read using a spectrophotometer. Sperm viability after thawing was assessed for each bull using a live/dead sperm viability kit (Year 1 = 3 replicates; Year 2 = 4 replicates). A minimum of 250 cells were assessed per bull in each replicate and classified as either live or dead. Finally, the ability of sperm to fertilize oocytes in vitro and their ability to develop to blastocyst stage embryos were assessed (5 replicates in each year involving 220 to 306 oocytes per bull). Data transformation to normalize residuals was required for mucus sperm penetration (square root) and IVF (cleavage and blastocyst rate) results (arcsin). The mean number of sperm counted at each 10 mm mark between 40 and 80 mm was higher in the high fertility (56.0; 95% CI 39.5 to 75.3) compared to the low fertility (42.9; 95% CI 29.3 to 59.1) Holstein Friesian bulls but the difference did not

  18. Antidermatophytic and Protease-inhibiting Activities of Zerumbone: A Natural Sesquiterpene from the Rhizome of Zingiber zerumbet (L.) Roscoe ex J.E; Smith

    PubMed Central

    Jyothilakshmi, Madhavankutty; Jyothis, Mathew; Narayanan, Gokulanathan Nair Hari; Latha, Mukalel Sankunni

    2017-01-01

    the development of new antidermatophytic drug.Cytotoxicity of zerumbone against Chang liver cell line was found to be very low compared with the reference drug doxorubicin. SUMMARY Zerumbone isolated from the rhizome of Zingiber zerumbet exhibited antidermatophytic activity against E. floccosum and M. canis (MIC 8 mg/L) and M. gypseum and T. Rubrum (MIC 16 mg/L).Zerumbone exhibited remarkable protease-inhibiting activity.Zerumbone is a novel candidate for the development of new antidermatophytic drug.Cytotoxicity of zerumbone against Chang liver cell line was found to be very low compared with the reference drug doxorubicin. Abbreviations used: CFU: colony forming unit, CLSI: Clinical and Laboratory Standards Institute, COX: cyclooxygenase, DMSO: dimethyl sulphoxide, EDTA: ethylene diamine tetra acetic acid, FT-IR: Fourier transform–infra red spectroscopy, HPLC: high-performance liquid chromatography, LOX: lipoxygenase, IMTECH: Institute of Microbial Technology, LCMS: liquid chromatography mass spectrometry, MTT: 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, MTCC: microbial type culture collection, MFC: minimum fungicidal concentration, MIC: minimum inhibitory concentration, MPO: myeloperoxidase, NMR: nuclear magnetic resonance spectroscopy, PAR: proteinase-activated receptor, PBS: phosphate-buffered saline, TCA: trichloro acetic acid PMID:28216875

  19. Assessment of cosmetic ingredients in the in vitro reconstructed human epidermis test method EpiSkin™ using HPLC/UPLC-spectrophotometry in the MTT-reduction assay.

    PubMed

    Alépée, N; Hibatallah, J; Klaric, M; Mewes, K R; Pfannenbecker, U; McNamee, P

    2016-06-01

    Cosmetics Europe recently established HPLC/UPLC-spectrophotometry as a suitable alternative endpoint detection system for measurement of formazan in the MTT-reduction assay of reconstructed human tissue test methods irrespective of the test system involved. This addressed a known limitation for such test methods that use optical density for measurement of formazan and may be incompatible for evaluation of strong MTT reducer and/or coloured chemicals. To build on the original project, Cosmetics Europe has undertaken a second study that focuses on evaluation of chemicals with functionalities relevant to cosmetic products. Such chemicals were primarily identified from the Scientific Committee on Consumer Safety (SCCS) 2010 memorandum (addendum) on the in vitro test EpiSkin™ for skin irritation testing. Fifty test items were evaluated in which both standard photometry and HPLC/UPLC-spectrophotometry were used for endpoint detection. The results obtained in this study: 1) provide further support for Within Laboratory Reproducibility of HPLC-UPLC-spectrophotometry for measurement of formazan; 2) demonstrate, through use a case study with Basazol C Blue pr. 8056, that HPLC/UPLC-spectrophotometry enables determination of an in vitro classification even when this is not possible using standard photometry and 3) addresses the question raised by SCCS in their 2010 memorandum (addendum) to consider an endpoint detection system not involving optical density quantification in in vitro reconstructed human epidermis skin irritation test methods.

  20. Derivation of an occupational exposure limit (OEL) for n-propyl bromide using an improved methodology.

    PubMed

    Rozman, Karl K; Doull, John

    2002-10-01

    n-Propyl bromide is an industrial solvent with increasing production volume due to its use as a replacement for fluorohydrocarbons. Therefore, the number of occupationally exposed workers is growing accordingly. This manuscript presents a thorough evaluation of available animal and human data to derive an occupational exposure limit (OEL) for n-propyl bromide. In addition, structure activity relationship within the homologous series of methyl, ethyl, and n-propyl bromide and an identical spectrum of effects caused by similar doses of 2-propyl bromide are used to increase the confidence of the analysis. The structure activity relationship was entirely consistent for acute and subchronic (neurologic, reproductive, and hematopoietic) toxicities and for mutagenic potency in that CH3Br was more toxic than CH3CH2Br, which in turn was more toxic than CH3CH2CH2Br in every case in all species studied, including humans. Animals appeared to be similarly susceptible as, or slightly more susceptible than, humans to n-propyl bromide's toxicity. An OEL (60-90 ppm) was derived from a limited human study and supported by an across-the-toxic-spectrum comparison of animal and human data for both n-propyl and 2-propyl bromide. A carcinogenic classification was not deemed necessary at the recommended OEL based on very low mutagenic potency and the consistent structure activity relationship across the homologous series of these alkyl bromides.

  1. Efficacy and safety of ipratropium bromide/albuterol delivered via Respimat inhaler versus MDI.

    PubMed

    Zuwallack, R; De Salvo, M C; Kaelin, T; Bateman, E D; Park, C S; Abrahams, R; Fakih, F; Sachs, P; Pudi, K; Zhao, Y; Wood, C C

    2010-08-01

    We compared the efficacy and safety of ipratropium bromide/albuterol delivered via Respimat inhaler, a novel propellant-free inhaler, versus chlorofluorocarbon (CFC)-metered dose inhaler (MDI) and ipratropium Respimat inhaler in patients with COPD. This was a multinational, randomized, double-blind, double-dummy, 12-week, parallel-group, active-controlled study. Patients with moderate to severe COPD were randomized to ipratropium bromide/albuterol (20/100mcg) Respimat inhaler, ipratropium bromide/albuterol MDI [36mcg/206mcg (Combivent Inhalation Aerosol MDI)], or ipratropium bromide (20mcg) Respimat inhaler. Each medication was administered four times daily. Serial spirometry was performed over 6h (0.15min, then hourly) on 4 test days. The primary efficacy variable was forced expiratory volume in 1s (FEV(1)) change from test day baseline at 12 weeks. A total of 1209 of 1480 randomized, treated patients completed the study; the majority were male (65%) with a mean age of 64 yrs and a mean screening pre-bronchodilator FEV(1) (percent predicted) of 41%. Ipratropium bromide/albuterol Respimat inhaler had comparable efficacy to ipratropium bromide/albuterol MDI for FEV(1) area under the curve at 0-6h (AUC(0-6)), superior efficacy to ipratropium Respimat inhaler for FEV(1) AUC(0-4) and comparable efficacy to ipratropium Respimat inhaler for FEV(1) AUC(4-6). All active treatments were well tolerated. This study demonstrates that ipratropium bromide/albuterol 20/100mcg inhaler administered four times daily for 12 weeks had equivalent bronchodilator efficacy and comparable safety to ipratropium bromide/albuterol 36mcg/206mcg MDI, and significantly improved lung function compared with the mono-component ipratropium bromide 20 mcg Respimat inhaler. [Clinical Trial Identifier Number: NCT00400153].

  2. The role of tiotropium bromide, a long-acting anticholinergic bronchodilator, in the management of COPD.

    PubMed

    Saberi, Farzad; O'Donnell, Denis E

    2005-01-01

    Bronchodilator therapy forms the mainstay of treatment for symptomatic patients with COPD. Long-acting bronchodilators, which maintain sustained airway patency over a 24-hour period, represent an advance in therapy. Tiotropium bromide is a new long-acting inhaled anticholinergic agent with superior pharmacodynamic properties compared with the short-acting anticholinergic, ipratropium bromide. Tiotropium bromide has been consistently shown to have a greater impact than ipratropium bromide on clinically important outcome measures such as health status. The mechanisms of clinical benefit with tiotropium bromide are multifactorial, but improved airway function, which enhances lung emptying and allows sustained deflation of over-inflated lungs, appears to explain improvements in dyspnea and exercise endurance in COPD. Inhaled tiotropium bromide therapy has also been associated with reduction in acute exacerbations of COPD as well as reduced hospitalizations. The safety profile of tiotropium bromide is impressive: dry mouth is the most common adverse event and rarely necessitates termination of the drug. No tachyphylaxis to tiotropium bromide has been demonstrated in clinical trials lasting up to 1 year. There is preliminary information that the combination of long-acting anticholinergics and long-acting beta2-adrenoceptor agonists provides additive physiological and clinical benefits. According to recent international guidelines, long-acting bronchodilators should be considered early in the management of symptomatic patients with COPD in order to achieve effective symptom alleviation and reduction in activity limitation. Tiotropium bromide, because of its once-daily administration and its established efficacy and tolerability profile, has emerged as an attractive therapeutic option for this condition.

  3. Rapid, easy cyanation of aryl bromides and chlorides using nickel salts in conjunction with microwave promotion.

    PubMed

    Arvela, Riina K; Leadbeater, Nicholas E

    2003-11-14

    We report here a fast, easy, and efficient method for the preparation of aryl nitriles from aryl bromides and chlorides. The methodology for aryl bromides involves the use of either Ni(CN)(2) or NaCN and NiBr(2). With aryl chlorides, a mix of NaCN and NiBr(2) is used and the reaction proceeds via the in situ formation of the corresponding aryl bromide. The reaction can be performed in air and is complete within 10 min.

  4. Bronchodilating effect of combined therapy with ipratropium bromide and ondansetron in patients with COPD.

    PubMed

    Pauwels, Romain; Joos, Guy F; Fogarty, Charles; Faiferman, Isidore; Hirschberg, Sandra; Vessey, Rupert; Wouters, Emiel F M

    2008-01-01

    The objectives of this study were to determine the effect of single and repeat dosing with oral ondansetron, a 5-HT3-specific receptor blocker, on the degree and duration of bronchodilation induced by inhaled ipratropium bromide in patients with COPD. Five clinics and university medical centers in four countries participated in the study; 47 patients with COPD were randomized to treatment; 44 completed all treatments. Patients had a baseline (pre-bronchodilator) FEV1>1L and post-bronchodilator (200 mcg salbutamol) FEV1<90% of predicted, with FEV1 reversibility (to 80 mcg inhaled ipratropium bromide and 400 mcg salbutamol) of at least 12% or 200 mL over baseline. The study was divided into two parts. In Part A, each patient received in a random order, four-way crossover manner, single doses of ondansetron placebo (oral) plus ipratropium bromide placebo (inhaled), ondansetron placebo plus ipratropium bromide 40 mcg inhaled via MDI, ondansetron 24 mg oral plus ipratropium bromide placebo and ondansetron 24 mg plus ipratropium bromide 40 mcg. In Part B, each patient received in a random order, two-way crossover manner, ipratropium bromide 40 mcg tid via MDI plus ondansetron 8 mg oral, qid, for 2 days; on day 3 patients received a single dose of ipratropium bromide 40 mcg plus 8 mg oral ondansetron. Alternatively, patients received ipratropium bromide via MDI and oral ondansetron placebo, as described above. Statistically significant differences in weighted mean FEV1 (0-6h), peak FEV1 and FEV1 determined 6h post-dose were noted comparing ipratropium bromide to placebo. Similar positive results were observed for sGaw and FVC. Addition of ondansetron to ipratropium bromide did not significantly modify values obtained with ipratropium alone. Ipratropium bromide induced a marked bronchodilation, compared to placebo. Addition of ondansetron (single or repeated doses) did not significantly increase the degree or duration of bronchodilation induced by ipratropium alone. s

  5. Locations of Bromide Ions in Tetragonal Lysozyme Crystals

    NASA Technical Reports Server (NTRS)

    Lim, Kap; Nadarajah, Arunan; Forsythe, Elizabeth L.; Pusey, Marc L.

    1998-01-01

    Anions have been shown to play a dominant role in the crystallization of chicken egg-white lysozyme from salt solutions. Previous studies employing X-ray crystallography have found one chloride ion binding site in the tetragonal crystal form of the protein and four nitrate ion binding sites in the monoclinic form. In this study the anion positions in the tetragonal form were determined from the difference Fourier map obtained from lysozyme crystals grown in bromide and chloride solutions. Five possible anion-binding sites were found in this manner. Some of these sites were in pockets containing basic residues while others were near neutral, but polar, residues. The sole chloride ion binding site found in previous studies was confirmed, while four further sites were found which corresponded to the four binding sites found for nitrate ions in monoclinic crystals. The study suggests that most of the anion-binding sites in lysozyme remain unchanged even when different anions and different crystal forms of lysozyme are employed.

  6. Structural and electronic properties of Diisopropylammonium bromide molecular ferroelectric crystal

    NASA Astrophysics Data System (ADS)

    Alsaad, A.; Qattan, I. A.; Ahmad, A. A.; Al-Aqtash, N.; Sabirianov, R. F.

    2015-10-01

    We report the results of ab-initio calculations based on Generalized Gradient Approximation (GGA) and hybrid functional (HSE06) of electronic band structure, density of states and partial density of states to get a deep insight into structural and electronic properties of P21 ferroelectric phase of Diisopropylammonium Bromide molecular crystal (DIPAB). We found that the optical band gap of the polar phase of DIPAB is ∼ 5 eV confirming it as a good dielectric. Examination of the density of states and partial density of states reveal that the valence band maximum is mainly composed of bromine 4p orbitals and the conduction band minimum is dominated by carbon 2p, carbon 2s, and nitrogen 2s orbitals. A unique aspect of P21 ferroelectric phase is the permanent dipole within the material. We found that P21 DIPAB has a spontaneous polarization of 22.64 consistent with recent findings which make it good candidate for the creation of ferroelectric tunneling junctions (FTJs) which have the potential to be used as memory devices.

  7. Factors affecting a cyanogen bromide-based assay of thiamin.

    PubMed

    Wyatt, D T; Lee, M; Hillman, R E

    1989-11-01

    We analyzed extensively a modified thiochrome method for thiamin analysis. Acid phosphatase (EC 3.1.3.2) from potato was superior to either alpha-amylase or acid phosphatase from wheat germ as a dephosphorylating agent. Timing of cyanogen bromide exposure was important, but the assay had good precision and accuracy. The standard curve was linear from 10 to 3000 nmol/L. The within-run and between-run coefficients of variation for total thiamin in whole blood were 3.6% and 7.4%, respectively. Analytical recoveries for low, intermediate, and high additions of thiamin to whole blood were 93-109%. Sample yield was increased by 41% (+/- 29% SD) with pre-assay freezing. Samples were stable for two days at room temperature, for seven days when refrigerated, and for two years when frozen. Previously unreported interference was seen with penicillin derivatives, and with several commonly used diuretic and antiepileptic medications. This assay may be suitable for population screening; 200 samples could be analyzed weekly at a cost of +0.20 per sample.

  8. Bioreactors for removing methyl bromide following contained fumigations

    USGS Publications Warehouse

    Miller, L.G.; Baesman, S.M.; Oremland, R.S.

    2003-01-01

    Use of methyl bromide (MeBr) as a quarantine, commodity, or structural fumigant is under scrutiny because its release to the atmosphere contributes to the depletion of stratospheric ozone. A closed-system bioreactor consisting of 0.5 L of a growing culture of a previously described bacterium, strain IMB-1, removed MeBr (> 110 ??mol L-1) from recirculating air. Strain IMB-1 grew slowly to high cell densities in the bioreactor using MeBr as its sole carbon and energy source. Bacterial oxidation of MeBr produced CO2 and hydrobromic acid (HBr), which required continuous neutralization with NaOH for the system to operate effectively. Strain IMB-1 was capable of sustained oxidation of large amounts of MeBr (170 mmol in 46 d). In an open-system bioreactor (10-L fermenter), strain IMB-1 oxidized a continuous supply of MeBr (220 ??mol L-1 in air). Growth was continuous, and 0.5 mol of MeBr was removed from the air supply in 14 d. The specific rate of MeBr oxidation was 7 ?? 10-16 mol cell-1 h-1. Bioreactors such as these can therefore be used to remove large quantities of contaminant MeBr, which opens the possibility of biodegradation as a practical means for its disposal.

  9. Bacterial oxidation of methyl bromide in Mono Lake, California

    USGS Publications Warehouse

    Connell, T.L.; Joye, S.B.; Miller, L.G.; Oremland, R.S.

    1997-01-01

    The oxidation of methyl bromide (MeBr) in the water column of Mono Lake, CA, was studied by measuring the formation of H14CO3 from [14C]MeBr. Potential oxidation was detected throughout the water column, with highest rates occurring in the epilimnion (5-12 m depth). The oxidation of MeBr was eliminated by filter-sterilization, thereby demonstrating the involvement of bacteria. Vertical profiles of MeBr activity differed from those obtained for nitrification and methane oxidation, indicating that MeBr oxidation is not simply a co-oxidation process by either nitrifiers or methanotrophs. Furthermore, specific inhibitors of methane oxidation and/or nitrification (e.g., methyl fluoride, acetylene, allyl sulfide) had no effect upon the rate of MeBr oxidation in live samples. Of a variety of potential electron donors added to Mono Lake water, only trimethylamine resulted in the stimulation of MeBr oxidation. Cumulatively, these results suggest that the oxidation of MeBr in Mono Lake waters is attributable to trimethylamine-degrading methylotrophs. Neither methyl chloride nor methanol inhibited the oxidation of [14C]MeBr in live samples, indicating that these bacteria directly oxidized MeBr rather than the products of MeBr nucleophilic substitution reactions.

  10. High brightness formamidinium lead bromide perovskite nanocrystal light emitting devices.

    PubMed

    Perumal, Ajay; Shendre, Sushant; Li, Mingjie; Tay, Yong Kang Eugene; Sharma, Vijay Kumar; Chen, Shi; Wei, Zhanhua; Liu, Qing; Gao, Yuan; Buenconsejo, Pio John S; Tan, Swee Tiam; Gan, Chee Lip; Xiong, Qihua; Sum, Tze Chien; Demir, Hilmi Volkan

    2016-11-09

    Formamidinium lead halide (FAPbX3) has attracted greater attention and is more prominent recently in photovoltaic devices due to its broad absorption and higher thermal stability in comparison to more popular methylammonium lead halide MAPbX3. Herein, a simple and highly reproducible room temperature synthesis of device grade high quality formamidinium lead bromide CH(NH2)2PbBr3 (FAPbBr3) colloidal nanocrystals (NC) having high photoluminescence quantum efficiency (PLQE) of 55-65% is reported. In addition, we demonstrate high brightness perovskite light emitting device (Pe-LED) with these FAPbBr3 perovskite NC thin film using 2,2',2″-(1,3,5-Benzinetriyl)-tris(1-phenyl-1-H-benzimidazole) commonly known as TPBi and 4,6-Bis(3,5-di(pyridin-3-yl)phenyl)-2-methylpyrimidine (B3PYMPM) as electron transport layers (ETL). The Pe-LED device with B3PYMPM as ETL has bright electroluminescence of up to 2714 cd/m(2), while the Pe-LED device with TPBi as ETL has higher peak luminous efficiency of 6.4 cd/A and peak luminous power efficiency of 5.7 lm/W. To our knowledge this is the first report on high brightness light emitting device based on CH(NH2)2PbBr3 widely known as FAPbBr3 nanocrystals in literature.

  11. Chloride, bromide and iodide scintillators with europium doping

    DOEpatents

    Zhuravleva, Mariya; Yang, Kan

    2014-08-26

    A halide scintillator material is disclosed where the halide may comprise chloride, bromide or iodide. The material is single-crystalline and has a composition of the general formula ABX.sub.3 where A is an alkali, B is an alkali earth and X is a halide which general composition was investigated. In particular, crystals of the formula ACa.sub.1-yEu.sub.yI.sub.3 where A=K, Rb and Cs were formed as well as crystals of the formula CsA.sub.1-yEu.sub.yX.sub.3 (where A=Ca, Sr, Ba, or a combination thereof and X=Cl, Br or I or a combination thereof) with divalent Europium doping where 0.ltoreq.y.ltoreq.1, and more particularly Eu doping has been studied at one to ten mol %. The disclosed scintillator materials are suitable for making scintillation detectors used in applications such as medical imaging and homeland security.

  12. Water-lithium bromide double-effect absorption cooling analysis

    NASA Astrophysics Data System (ADS)

    Vliet, G. C.; Lawson, M. B.; Lithgow, R. A.

    1980-12-01

    A numerical model was developed for the transient simulation of the double-effect, water-lithium bromide absorption cooling machine and was used to determine the effect of the various design and input variables on the absorption unit performance. The performance parameters considered were coefficient of performance and cooling capacity. The variables considered include source hot water, cooling water, and chilled water temperatures; source hot water, cooling water, and chilled water flow rates; solution circulation rate; heat exchanger areas; pressure drop between evaporator and absorber; solution pump characteristics; and refrigerant flow control methods. The performance sensitivity study indicates that the distribution of heat exchanger area among the various (seven) heat exchange components is a very important design consideration. Moreover, it indicated that the method of flow control of the first effect refrigerant vapor through the second effect is a critical design feature when absorption units operate over a significant range of cooling capacity. The model was used to predict the performance of the Trane absorption unit with fairly good accuracy.

  13. [Treatment of cetyltrimethyl ammonium bromide wastewater by potassium ferrate].

    PubMed

    Yang, Wei-hua; Wang, Hong-hui; Zeng, Xiao-xu; Huang, Ting-ting

    2009-08-15

    A novel oxidant potassium ferrate (K2FeO4) was used to remove cetyltrimethyl ammonium bromide (CTAB) at room temperature. The effects of various conditions on the removal ratio, such as reaction time, dosing quantity of K2FeO4 and initial pH, were investigated. The experiments results show that the removal ratio reaches 79.4% when the reaction time is 30 min, the dosing quantity of K2FeO4 to CTAB is 1:1, the initial pH of the solution is 7. In the reaction progress, the oxidation of K2FeO4 and the flocculation of the reduction product have synergistic effect on the removal of CTAB. In addition, infrared spectra of CTAB before and after being treated with K2FeO4 were further studied. The results indicate that the degradation process involves the interruption of chain and the subsequent mineralization to inorganic molecules. Furthermore, the reaction of K2FeO4 and CTAB follows second order kinetics law.

  14. Reactive films for mitigating methyl bromide emissions from fumigated soil.

    PubMed

    Xuan, Richeng; Ashworth, Daniel J; Luo, Lifang; Yates, Scott R

    2011-03-15

    Emissions of methyl bromide (MeBr) from agricultural fumigation can lead to depletion of the stratospheric ozone layer, and so its use is being phased out. However, as MeBr is still widely used under Critical Use Exemptions, strategies are still required to control such emissions. In this work, novel reactive films (RFs) were designed and their efficacy in limiting loss of MeBr from soil was tested. A reactive layer, containing dry ammonium thiosulfate (ATS), was sandwiched between two layers of plastic film, the lower layer being HDPE (high-density polyethylene film, which is permeable to MeBr) and the upper layer HDPE or VIF (virtually impermeable film). MeBr diffusion through, and transformation by, the RFs were tested in a stainless-steel permeability cell. Although ineffective when dry, the RFs substantially depleted MeBr when activated with water to produce ATS solution. MeBr half-life (t(1/2)) was around 9.0 h at 20 °C in the presence of activated RF, and was sensitive to temperature (t(1/2) 15.7 and 2.9 h at 10 and 40 °C, respectively). When the upper film layer was VIF, less than 0.15% of the added MeBr diffused through the film, with the remainder being transformed within the reactive layer. These findings indicate that such films have good potential to reduce MeBr loss from fumigated soils to the atmosphere.

  15. Evaluation of Alkali Bromide Salts for Potential Pyrochemical Applications

    SciTech Connect

    Prabhat K. Tripathy; Steven D. Herrmann; Guy L. Fredrickson; Tedd E. Lister; Toni Y. Gutknecht

    2013-10-01

    Transient techniques were employed to study the electrochemical behavior, reduction mechanism and transport properties of REBr3 (RE - La, Nd and Gd) in pure LiBr, LiBr-KBr (eutectic) and LiBr-KBr-CsBr (eutectic) melts. Gd(III) showed a reversible single step soluble-insoluble exchange phenomenon in LiBr melt at 973K. Although La (III), Nd(III) and Gd(III) ions showed reversible behavior in eutectic LiBr-KBr melts, these ions showed a combination of temperature dependent reversible and pseudo-reversible behavior. While both La(III) and Gd(III) showed one step reduction, the reduction of Nd(III) was observed to be a two step process. La metal could be electrodeposited from the ternary electrolyte at a temperature of 673K. Various electrochemical measurements suggest that both binary and ternary bromide melts can potentially be used to electrodeposit high purity RE metals at comparatively lower operating temperatures.

  16. Associated anisotropy decays of ethidium bromide interacting with DNA

    NASA Astrophysics Data System (ADS)

    Chib, Rahul; Raut, Sangram; Sabnis, Sarika; Singhal, Preeti; Gryczynski, Zygmunt; Gryczynski, Ignacy

    2014-03-01

    Ethidium Bromide (EB) is a commonly used dye in a deoxyribonucleic acid (DNA) study. Upon an intercalation, this dye significantly increases its brightness and fluorescence lifetime. In this report we have studied the time resolved fluorescence properties of EB existing simultaneously in free and DNA-bound forms in the solution. Fluorescence intensity decays were fitted globally to a double exponential model with lifetimes corresponding to free (1.6 ns) and bound (22 ns) forms, and molar fractions were determined for all used solutions. Anisotropy decays displayed characteristic time dependence with an initial rapid decline followed by recovery and slow decay. The short-lived fraction associated with free EB molecules decreases faster than long-lived fraction associated with EB bound to DNA. Consequently, contribution from fast rotation leads to initial rapid decay in anisotropy. On the other hand bound fraction, due to slow rotation helps recover anisotropy in time. This effect of associated anisotropy decays in systems such as EB free/EB-DNA is clearly visible in a wide range of concentrations, and should be taken into account in polarization assays and biomolecule dynamics studies.

  17. High brightness formamidinium lead bromide perovskite nanocrystal light emitting devices

    PubMed Central

    Perumal, Ajay; Shendre, Sushant; Li, Mingjie; Tay, Yong Kang Eugene; Sharma, Vijay Kumar; Chen, Shi; Wei, Zhanhua; Liu, Qing; Gao, Yuan; Buenconsejo, Pio John S.; Tan, Swee Tiam; Gan, Chee Lip; Xiong, Qihua; Sum, Tze Chien; Demir, Hilmi Volkan

    2016-01-01

    Formamidinium lead halide (FAPbX3) has attracted greater attention and is more prominent recently in photovoltaic devices due to its broad absorption and higher thermal stability in comparison to more popular methylammonium lead halide MAPbX3. Herein, a simple and highly reproducible room temperature synthesis of device grade high quality formamidinium lead bromide CH(NH2)2PbBr3 (FAPbBr3) colloidal nanocrystals (NC) having high photoluminescence quantum efficiency (PLQE) of 55–65% is reported. In addition, we demonstrate high brightness perovskite light emitting device (Pe-LED) with these FAPbBr3 perovskite NC thin film using 2,2′,2″-(1,3,5-Benzinetriyl)-tris(1-phenyl-1-H-benzimidazole) commonly known as TPBi and 4,6-Bis(3,5-di(pyridin-3-yl)phenyl)-2-methylpyrimidine (B3PYMPM) as electron transport layers (ETL). The Pe-LED device with B3PYMPM as ETL has bright electroluminescence of up to 2714 cd/m2, while the Pe-LED device with TPBi as ETL has higher peak luminous efficiency of 6.4 cd/A and peak luminous power efficiency of 5.7 lm/W. To our knowledge this is the first report on high brightness light emitting device based on CH(NH2)2PbBr3 widely known as FAPbBr3 nanocrystals in literature. PMID:27827424

  18. High brightness formamidinium lead bromide perovskite nanocrystal light emitting devices

    NASA Astrophysics Data System (ADS)

    Perumal, Ajay; Shendre, Sushant; Li, Mingjie; Tay, Yong Kang Eugene; Sharma, Vijay Kumar; Chen, Shi; Wei, Zhanhua; Liu, Qing; Gao, Yuan; Buenconsejo, Pio John S.; Tan, Swee Tiam; Gan, Chee Lip; Xiong, Qihua; Sum, Tze Chien; Demir, Hilmi Volkan

    2016-11-01

    Formamidinium lead halide (FAPbX3) has attracted greater attention and is more prominent recently in photovoltaic devices due to its broad absorption and higher thermal stability in comparison to more popular methylammonium lead halide MAPbX3. Herein, a simple and highly reproducible room temperature synthesis of device grade high quality formamidinium lead bromide CH(NH2)2PbBr3 (FAPbBr3) colloidal nanocrystals (NC) having high photoluminescence quantum efficiency (PLQE) of 55–65% is reported. In addition, we demonstrate high brightness perovskite light emitting device (Pe-LED) with these FAPbBr3 perovskite NC thin film using 2,2‧,2″-(1,3,5-Benzinetriyl)-tris(1-phenyl-1-H-benzimidazole) commonly known as TPBi and 4,6-Bis(3,5-di(pyridin-3-yl)phenyl)-2-methylpyrimidine (B3PYMPM) as electron transport layers (ETL). The Pe-LED device with B3PYMPM as ETL has bright electroluminescence of up to 2714 cd/m2, while the Pe-LED device with TPBi as ETL has higher peak luminous efficiency of 6.4 cd/A and peak luminous power efficiency of 5.7 lm/W. To our knowledge this is the first report on high brightness light emitting device based on CH(NH2)2PbBr3 widely known as FAPbBr3 nanocrystals in literature.

  19. CHLORPYRIFOS TRANSFORMATION BY AQUEOUS CHLORINE IN THE PRESENCE OF BROMIDE AND NATURAL ORGANIC MATTER

    EPA Science Inventory

    The aqueous chlorination of chlorpyrifos (CP) was investigated in the presence of bromide and natural organic matter (NOM), which were identified as naturally occurring aqueous constituents that could impact CP transformation rates to the toxic product chlorpyrifos oxon (CPO). Br...

  20. Palladium-catalyzed carbonylation reaction of aryl bromides with 2-hydroxyacetophenones to form flavones.

    PubMed

    Wu, Xiao-Feng; Neumann, Helfried; Beller, Matthias

    2012-10-01

    Flavone of the month: a general and efficient method for the palladium-catalyzed carbonylative synthesis of flavones has been developed. Starting from aryl bromides and 2-hydroxyacetophenones, the corresponding flavones have been isolated in good yields.

  1. BROMIDE'S EFFECT ON DBP FORMATION, SPECIATION, AND CONTROL: PART 1, OZONATION

    EPA Science Inventory

    The effect of variable ozone dosage and bromide concentration on the formation of organic disinfection byproducts (DBPs) and bromate were evaluated. Low ozone dosages resulted in oxidation of organic precursors, yielding decreases in the formation potential for total trihalometha...

  2. Stabilized thallium bromide radiation detectors and methods of making the same

    SciTech Connect

    Leao, Cedric Rocha; Lordi, Vincenzo

    2015-11-24

    According to one embodiment, a crystal includes thallium bromide (TlBr), one or more positively charged dopants, and one or more negatively charged dopants. According to another embodiment, a system includes a monolithic crystal including thallium bromide (TlBr), one or more positively charged dopants, and one or more negatively charged dopants; and a detector configured to detect a signal response of the crystal.

  3. Influence of bromide on the performance of the amphipod Hyalella azteca in reconstituted waters

    USGS Publications Warehouse

    Ivey, Chris D.; Ingersoll, Christopher G.

    2016-01-01

    Poor performance of the amphipod Hyalella azteca has been observed in exposures using reconstituted waters. Previous studies have reported success in H. azteca water-only exposures with the addition of relatively high concentrations of bromide. The present study evaluated the influence of lower environmentally representative concentrations of bromide on the response ofH. azteca in 42-d water-only exposures. Improved performance of H. azteca was observed in reconstituted waters with >0.02 mg Br/L.

  4. Small airways ventilation heterogeneity and hyperinflation in COPD: response to tiotropium bromide.

    PubMed

    Verbanck, Sylvia; Schuermans, Daniël; Vincken, Walter

    2007-01-01

    In chronic obstructive pulmonary disease (COPD) patients tiotropium bromide has been shown to improve forced expiratory volume in one second (FEV1) and inspiratory capacity (IC). We investigated whether the mechanism leading to these improvements is related to small airways ventilation heterogeneity, assessed by multiple breath washout tests. Forty stable tiotropium-free COPD patients (FEV1: 27%-78% predicted) were studied before and 90 min after administration of tiotropium bromide on visit0, and following 3 and 6 weeks of tiotropium bromide treatment (visit3wks, visit6wks). After study completion, COPD patients were classified into two subgroups according to degree of hyperinflation at visit0 (Hyp-, Hyp+). The Hyp+ group showed significant increases in trough (ie, pre-dose) FEV1 and IC after 3 and 6 weeks of tiotropium bromide, and the 90 min tiotropium bromide responses of FEV1 and IC were significant at visit0 (p < or = 0.001 for both) but not during subsequent visits. The Hyp- group showed significant FEV1 increases 90 min after tiotropium bromide on all three visits (all p < 0.005) but no sustained increase in trough values. In both COPD subgroups, the grossly abnormal ventilation heterogeneity barely showed any significant improvements with tiotropium bromide in the conductive airways (without changes in trough value) and no changes at all in the acinar airways. We conclude that the sustained improvements in trough IC and FEV1 with tiotropium bromide predominantly observed in COPD patients with considerable hyperinflation, are unrelated to small airways ventilation heterogeneity.

  5. Flow Boiling Heat Transfer to Lithium Bromide Aqueous Solution in Subcooled Region

    NASA Astrophysics Data System (ADS)

    Furukawa, Masahiro; Kaji, Masao; Nishizumi, Takeharu; Ozaki, Shinji; Sekoguchi, Kotohiko

    To improve the thermal performance of high temperature generator of absorption chiller/heater, heat transfer characteristics of flow boiling of lithium bromide aqueous solution in the subcooled region were experimentally investigated. Experiments were made for water and lithium bromide aqueous solution flowing in a rectangular channel (5 mm × 20 mm cross section) with one side wall heated. Boiling onset quality of lithium bromide aqueous solution is greater than that of water. The heat transfer coefficient of lithium bromide aqueous solution is about a half of that of water under the same experimental conditions of inlet velocity and heat flux. The experimental data of heat transfer coefficient for water are compared with the empirical correlation of Thom et al.11) and a fairly good agreement is obtained. The predictive calculations by the method of Sekoguchi et al.12) are compared with the data for water and lithium bromide aqueous solution. Agreement between them is good for water, while the results for lithium bromide aqueous solution are not satisfactory.

  6. Preferential bromide and pesticide movement to tile drains under different cropping practices.

    PubMed

    Fortin, J; Gagnon-Bertrand, E; Vézina, L; Rompré, M

    2002-01-01

    Subsurface drainage systems are useful tools to study chemical leaching in soils. Our objective was to compare the breakthrough behavior of bromide, atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-triazine) and metolachlor [2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl) acetamid] to tile drains under two fall tillage practices (conventional tillage [CT] with a moldboard plow, and reduced tillage [RT] with a chisel plow) in field plots cultivated with corn (Zea mays L.). Leachate volume were greater in RT than in CT, with no statistical differences. Soil analysis showed that bromide migrated deeper in the soil profile than both herbicides, with little tillage effect. All chemicals were detected in drainage water at the same time and followed an event-driven behavior. Tillage had no effect on atrazine and metolachlor found in drainage water, while bromide concentration peaks were higher in RT than in CT in 1999. Concentration peaks were recorded earlier for atrazine and metolachlor than for bromide. Plots of cumulative relative chemical mass (cumulative mass divided by total mass measured in drainage) as a function of cumulative drainage were mostly linear for bromide, while they were S-shaped for both herbicides. Drainage that corresponded to 50% of relative cumulative mass ranged from 40 to 55% for bromide and from 5 to 28% for both herbicides. Rapid chemical movement to tile drains suggested that preferential flow was important in both CT and RT, and that these tillage practices had little influence on this phenomena.

  7. Dicationic alkylammonium bromide gemini surfactants. Membrane perturbation and skin irritation.

    PubMed

    Almeida, João A S; Faneca, Henrique; Carvalho, Rui A; Marques, Eduardo F; Pais, Alberto A C C

    2011-01-01

    Dicationic alkylammonium bromide gemini surfactants represent a class of amphiphiles potentially effective as skin permeation enhancers. However, only a limited number of studies has been dedicated to the evaluation of the respective cytotoxicity, and none directed to skin irritation endpoints. Supported on a cell viability study, the cytotoxicity of gemini surfactants of variable tail and spacer length was assessed. For this purpose, keratinocyte cells from human skin (NCTC 2544 cell line), frequently used as a model for skin irritation, were employed. The impact of the different gemini surfactants on the permeability and morphology of model vesicles was additionally investigated by measuring the leakage of calcein fluorescent dye and analyzing the NMR spectra of ³¹P, respectively. Detail on the interaction of gemini molecules with model membranes was also provided by a systematic differential scanning calorimetry (DSC) and molecular dynamics (MD) simulation. An irreversible impact on the viability of the NCTC 2544 cell line was observed for gemini concentrations higher than 25 mM, while no cytotoxicity was found for any of the surfactants in a concentration range up to 10 mM. A higher cytotoxicity was also found for gemini surfactants presenting longer spacer and shorter tails. The same trend was obtained in the calorimetric and permeability studies, with the gemini of longest spacer promoting the highest degree of membrane destabilization. Additional structural and dynamical characterization of the various systems, obtained by ³¹P NMR and MD, provide some insight on the relationship between the architecture of gemini surfactants and the respective perturbation mechanism.

  8. Silica nanoparticles separation from water: aggregation by cetyltrimethylammonium bromide (CTAB).

    PubMed

    Liu, Y; Tourbin, M; Lachaize, S; Guiraud, P

    2013-07-01

    Nanoparticles will inevitably be found in industrial and domestic wastes in the near future and as a consequence soon in water resources. Due to their ultra-small size, nanoparticles may not only have new hazards for environment and human health, but also cause low separation efficiency by classical water treatments processes. Thus, it would be an important challenge to develop a specific treatment with suitable additives for recovery of nanoparticles from waters. For this propose, this paper presents aggregation of silica nanoparticles (Klebosol 30R50 (75nm) and 30R25 (30nm)) by cationic surfactant cetyltrimethylammonium bromide (CTAB). Different mechanisms such as charge neutralization, "depletion flocculation" or "volume-restriction", and "hydrophobic effect" between hydrocarbon tails of CTAB have been proposed to explicate aggregation results. One important finding is that for different volume concentrations between 0.05% and 0.51% of 30R50 suspensions, the same critical coagulation concentration was observed at CTAB=0.1mM, suggesting the optimized quantity of CTAB during the separation process for nanoparticles of about 75nm. Furthermore, very small quantities of CTAB (0.01mM) can make 30R25 nanosilica aggregated due to the "hydrophobic effect". It is then possible to minimize the sludge and allow the separation process as "greener" as possible by studying this case. It has also shown that aggregation mechanisms can be different for very small particles so that a special attention has to be paid to the treatment of nanoparticles contained in water and wastewaters.

  9. Dicationic Alkylammonium Bromide Gemini Surfactants. Membrane Perturbation and Skin Irritation

    PubMed Central

    Almeida, João A. S.; Faneca, Henrique; Carvalho, Rui A.; Marques, Eduardo F.; Pais, Alberto A. C. C.

    2011-01-01

    Dicationic alkylammonium bromide gemini surfactants represent a class of amphiphiles potentially effective as skin permeation enhancers. However, only a limited number of studies has been dedicated to the evaluation of the respective cytotoxicity, and none directed to skin irritation endpoints. Supported on a cell viability study, the cytotoxicity of gemini surfactants of variable tail and spacer length was assessed. For this purpose, keratinocyte cells from human skin (NCTC 2544 cell line), frequently used as a model for skin irritation, were employed. The impact of the different gemini surfactants on the permeability and morphology of model vesicles was additionally investigated by measuring the leakage of calcein fluorescent dye and analyzing the NMR spectra of 31P, respectively. Detail on the interaction of gemini molecules with model membranes was also provided by a systematic differential scanning calorimetry (DSC) and molecular dynamics (MD) simulation. An irreversible impact on the viability of the NCTC 2544 cell line was observed for gemini concentrations higher than 25 mM, while no cytotoxicity was found for any of the surfactants in a concentration range up to 10 mM. A higher cytotoxicity was also found for gemini surfactants presenting longer spacer and shorter tails. The same trend was obtained in the calorimetric and permeability studies, with the gemini of longest spacer promoting the highest degree of membrane destabilization. Additional structural and dynamical characterization of the various systems, obtained by 31P NMR and MD, provide some insight on the relationship between the architecture of gemini surfactants and the respective perturbation mechanism. PMID:22102870

  10. The fate of alternative soil fumigants to methyl bromide

    NASA Astrophysics Data System (ADS)

    Qin, R.; Gao, S.

    2011-12-01

    Soil fumigation is an important agricultural practice for the control of soil-borne pests. Since the phase-out of methyl bromide, due to its role in the depletion of stratospheric ozone, several alternatives such as 1,3-dichloropropene (1,3-D), chloropicrin (CP), and dimethyl disulfide (DMDS) are being increasingly used. The major processes and factors affecting the fate of these chemicals are evaluated. The high volatility of fumigants leads to high emission loss when no proper containment is used. Recent tarping technology using low permeability films can significantly reduce emissions. Fumigant degradation rate becomes critical to the determination of fumigation rate that affects efficacy and residence time in soil. A series of laboratory incubation experiments were carried out to determine degradation rate of 1,3-D isomers, CP and DMDS in five different soils collected from California and Florida. Fumigant degradation rates depend highly on the amounts of fumigants in soil, chemical characteristics, and soil conditions. Fumigant degradation rate were found to increase for all fumigants as the fumigant amounts in soil decreased. The changes were smaller for 1,3-D isomers compared to CP and DMDS. In soils with the lowest application rate, the degradation rate of fumigants is in the order of CP > DMDS > cis-1,3-D > trans-1,3-D. Soil and environmental factors also affect fumigant degradation rate. These findings suggest that a proper application rate should be determined for specific fumigants in a soil when using low permeability tarp in order to achieve sufficient fumigation efficacy during a certain period of time while minimizing potential surge of emissions after tarp removal and/or long residence time in soil that may cause phytotoxicity or leaching.

  11. Predicting bromide incorporation in a chlorinated indoor swimming pool.

    PubMed

    Chowdhury, Shakhawat; Mazumder, Abu Jafar; Husain, Tahir

    2016-06-01

    The water in and air above swimming pools often contain high levels of disinfection byproducts (DBPs) due to chemical reactions between chlorine- or bromine-based disinfectants and organic/inorganic matter in the source water and released from swimmers. Exposure to these DBPs, though inevitable, can pose health threats to humans. In this study, DBPs in tap water (S1), and water from a chlorinated indoor swimming pool before (S2) and after swimming (S3) were measured. The brominated species constituted the majority of DBPs formed in S1, S2, and S3. Trihalomethanes (THMs) in S3 was 6.9 (range 2.9-11.1) and 1.4 (range 0.52-2.9) times those in S1 and S2, respectively; and the haloacetic acids (HAAs) in S3 was 4.2 (range 2.5-7.5) and 1.2 (range 0.6-2.6) times those in S1 and S2, respectively. The mean THMs in air above the swimming pool before (S2-A) and after swimming (S3-A) were 72.2 and 93.0 μg/m(3), respectively, and their ranges were 36.3-105.8 and 44.1-133.6 μg/m(3), respectively. The average percentages of bromide incorporation (BI) into THMs in S1, S2, and S3 were 3.0, 9.3, and 10.6 %, respectively; and the BI into HAAs in S1, S2, and S3 were 6.6, 12.0, and 12.2 %, respectively. Several models were trained for predicting the BI into THMs and HAAs. The results indicate that additional information is required to develop predictive models for BI in swimming pools.

  12. Bacterial oxidation of methyl bromide in fumigated agricultural soils

    USGS Publications Warehouse

    Miller, L.G.; Connell, T.L.; Guidetti, J.R.; Oremland, R.S.

    1997-01-01

    The oxidation of [14C]methyl bromide ([14C]MeBr) to 14CO2 was measured in field experiments with soils collected from two strawberry plots fumigated with mixtures of MeBr and chloropicrin (CCI3NO2). Although these fumigants are considered potent biocides, we found that the highest rates of MeBr oxidation occurred 1 to 2 days after injection when the fields were tarped, rather than before or several days after injection. No oxidation of MeBr occurred in heat-killed soils, indicating that microbes were the causative agents of the oxidation. Degradation of MeBr by chemical and/or biological processes accounted for 20 to 50% of the loss of MeBr during fumigation, with evasion to the atmosphere inferred to comprise the remainder. In laboratory incubations, complete removal of [14C]MeBr occurred within a few days, with 47 to 67% of the added MeBr oxidized to 14CO2 and the remainder of counts associated with the solid phase. Chloropicrin inhibited the oxidation of MeBr, implying that use of this substance constrains the extent of microbial degradation of MeBr during fumigation. Oxidation was by direct bacterial attack of MeBr and not of methanol, a product of the chemical hydrolysis of MeBr. Neither nitrifying nor methane-oxidizing bacteria were sufficiently active in these soils to account for the observed oxidation of MeBr, nor could the microbial degradation of MeBr be linked to cooxidation with exogenously supplied electron donors. However, repeated addition of MeBr to live soils resulted in higher rates of its removal, suggesting that soil bacteria used MeBr as an electron donor for growth. To support this interpretation, we isolated a gram-negative, aerobic bacterium from these soils which grew with MeBr as a sole source of carbon and energy.

  13. Eudragit RS PO nanoparticles for sustained release of pyridostigmine bromide

    NASA Astrophysics Data System (ADS)

    Hoobakht, Fatemeh; Ganji, Fariba; Vasheghani-Farahani, Ebrahim; Mousavi, Seyyed Mohammad

    2013-09-01

    Pyridostigmine bromide (PB) is an inhibitor of cholinesterase, which is used in the treatment of myasthenia gravis and administered for protection against exposure to toxic nerve agents. Tests were done to investigate prolonging the half-life of PB and improving its release behavior. PB was loaded in nanoparticles (NPs) of Eudragit RS PO (Eu-RS) prepared using the technique of quasi emulsion solvent diffusion. Variables of output power of the sonicator, bath temperature and mixing time, were chosen as the optimization factors to obtain the minimum sized NPs. In addition, emulsions were tested at different ratios of drug-to-polymer by dynamic light scattering to determine size and zeta potential of NPs. UV-spectroscopy was used to determine PB content of the NPs. Drug-loaded NPs were characterized by scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectra. Results determined that mixing time had a significant impact on the size of Eu-RS NPs, but power output of sonicator and bath temperature had no significant effect. The particle size obtained at the optimum condition (power output of 70 W, bath temperature of 33 °C, and mixing time of 7 min) was less than 200 nm (optimum sizes were 138.9 and 179.5 nm for Eu-RS and PB-loaded Eu-RS NPs, respectively). The optimum PB-loaded Eu-RS NPs at the PB to Eu-RS weight ratio of 1-4 and 20 % of loaded PB released from the nanocarriers within 100 h.

  14. Clinical potential of aclidinium bromide in chronic obstructive pulmonary disease

    PubMed Central

    Jones, Paul W

    2015-01-01

    Three long-acting muscarinic antagonists (LAMAs) are now available in Europe, providing clinicians and patients with a choice of interventions, which is important in COPD, which is clinically a heterogeneous disease. The first LAMA, tiotropium, has been widely used over the last decade as a once-daily maintenance therapy in stable COPD to improve patients’ health-related quality of life and to reduce the risk of exacerbations. Administered via the HandiHaler® device, it is safe and well tolerated. Another new once-daily LAMA, glycopyrronium, has also been shown to improve health status and reduce exacerbations, and is well tolerated. The subject of this review is a third LAMA, aclidinium bromide, which was approved as a twice-daily maintenance bronchodilator treatment. In the pivotal Phase III clinical trials, patients receiving aclidinium achieved significantly greater improvements in lung function, reductions in breathlessness, and improvements in health status compared with placebo, for up to 24 weeks. In continuation studies, these improvements were sustained for up to 52 weeks. Pooled data showed exacerbation frequency was significantly reduced with aclidinium versus placebo. Preclinical and pharmacological studies demonstrating low systemic bioavailability and a low propensity to induce cardiac arrhythmias were translated into a favorable tolerability profile in the clinical trial program – the adverse event profile of aclidinium was similar to placebo, with a low incidence of anticholinergic and cardiac adverse events. While additional studies are needed to evaluate its full clinical potential, aclidinium is an important part of this recent expansion of LAMA therapeutic options, providing clinicians and patients with an effective and well-tolerated COPD treatment. PMID:25848244

  15. Carbon isotope fractionation of methyl bromide during agricultural soil fumigations

    USGS Publications Warehouse

    Bill, M.; Miller, L.G.; Goldstein, Allen H.

    2002-01-01

    The isotopic composition of methyl bromide (CH3Br) has been suggested to be a potentially useful tracer for constraining the global CH3Br budget. In order to determine the carbon isotopic composition of CH3Br emitted from the most significant anthropogenic application (pre-plant fumigation) we directly measured the ??13C of CH3Br released during commercial fumigation. We also measured the isotopic fractionation associated with degradation in agricultural soil under typical field fumigation conditions. The isotopic composition of CH3Br collected in soil several hours after injection of the fumigant was -44.5??? and this value increased to -20.7??? over the following three days. The mean kinetic isotope effect (KIE) associated with degradation of CH3Br in agricultural soil (12???) was smaller than the reported value for methylotrophic bacterial strain IMB-1, isolated from previously fumigated agricultural soil, but was similar to methylotrophic bacterial strain CC495, isolated from a pristine forest litter zone. Using this fractionation associated with the degradation of CH3Br in agricultural soil and the mean ??13C of the industrially manufactured CH3Br (-54.4???), we calculate that the agricultural soil fumigation source has a carbon isotope signature that ranges from -52.8??? to -42.0???. Roughly 65% of industrially manufactured CH3Br is used for field fumigations. The remaining 35% is used for structural and post-harvest fumigations with a minor amount used during industrial chemical manufacturing. Assuming that the structural and post-harvest fumigation sources of CH3Br are emitted without substantial fractionation, we calculate that the ??13C of anthropogenically emitted CH3Br ranges from -53.2??? to -47.5???.

  16. Inhalable Ipratropium Bromide Particle Engineering with Multicriteria Optimization.

    PubMed

    Vinjamuri, Bhavani Prasad; Haware, Rahul V; Stagner, William C

    2016-11-21

    Spray-dried ipratropium bromide (IPB) microspheres for oral inhalation were engineered using Quality by Design. The interrogation of material properties, process parameters, and critical product quality attributes interplay enabled rational product design. A 2(7-3) screening design exhibited the Maillard reaction between L-leucine (LL) and lactose at studied outlet temperatures (OT) >130°C. A response surface custom design was used in conjunction with multicriteria optimization to determine the operating design space to achieve inhalable microparticles. Statistically significant predictive models were developed for volume median diameter (p = 0.0001, adjusted R (2)  = 0.9938), span (p = 0.0278, adjusted R (2)  = 0.7912), yield (p = 0.0020, adjusted R (2)  = 0.9320), and OT (p = 0.0082, adjusted R (2)  = 0.8768). An independent verification batch confirmed the model's predictive capability. The prediction and actual values were in good agreement. Particle size and span were 3.32 ± 0.09 μm and 1.71 ± 0.18, which were 4.7 and 5.3% higher than the predicted values. The process yield was 50.3%, compared to the predicted value of 65.3%. The OT was 100°C versus the predicted value of 105°C. The label strength of IPB microparticles was 99.0 to 105.9% w/w suggesting that enrichment occurred during the spray-drying process. The present study can be utilized to initiate the design of the first commercial IPB dry powder inhaler.

  17. Depleting methyl bromide residues in soil by reaction with bases.

    PubMed

    Xuan, Richeng; Ashworth, Daniel J; Luo, Lifang; Wang, Haizhen; Yates, Scott R

    2010-12-01

    Despite generally being considered the most effective soil fumigant, methyl bromide (MeBr) use is being phased out because its emissions from soil can lead to stratospheric ozone depletion. However, a large amount is still currently used due to Critical Use Exemptions. As strategies for reducing the postfumigation emissions of MeBr from soil, Ca(OH)(2), K(2)CO(3), and NH(3) were assessed as means of promoting MeBr degradation. Ammonia aqueous solution (NH(4)OH) was the most effective, because MeBr can be degraded by both hydrolysis and ammonolysis. At 20 °C, the half-lives (t(1/2)) of MeBr were 18.0, 2.5, and 1.3 h in 0.1, 1.0, and 2.0 M NH(4)OH, respectively. In 1.0 M NH(4)OH, increasing the solution temperature to 40 °C reduced the half-life of MeBr to 0.23 h. Ammonia amendment to moist soil also promoted MeBr transformation, and the MeBr degradation rate increased with increasing soil temperature. NH(4)OH (30%, 16 M) very effectively reacted with MeBr that was contained under plastic film. Under Hytibar (a virtually impermeable film, VIF), over 99.5% of the MeBr could be destroyed by 30% NH(4)OH in 8 h at 20 °C. On the basis of these results, good management practices (i.e., VIF plus NH(4)OH) could be developed for continued use of MeBr as a soil fumigant under Critical Use Exemptions, without significant emissions.

  18. Naringenin-loaded solid lipid nanoparticles: preparation, controlled delivery, cellular uptake, and pulmonary pharmacokinetics.

    PubMed

    Ji, Peng; Yu, Tong; Liu, Ying; Jiang, Jie; Xu, Jie; Zhao, Ying; Hao, Yanna; Qiu, Yang; Zhao, Wenming; Wu, Chao

    2016-01-01

    Naringenin (NRG), a flavonoid compound, had been reported to exhibit extensive pharmacological effects, but its water solubility and oral bioavailability are only~46±6 µg/mL and 5.8%, respectively. The purpose of this study is to design and develop NRG-loaded solid lipid nanoparticles (NRG-SLNs) to provide prolonged and sustained drug release, with improved stability, involving nontoxic nanocarriers, and increase the bioavailability by means of pulmonary administration. Initially, a group contribution method was used to screen the best solid lipid matrix for the preparation of SLNs. NRG-SLNs were prepared by an emulsification and low-temperature solidification method and optimized using an orthogonal experiment approach. The morphology was examined by transmission electron microscopy, and the particle size and zeta potential were determined by photon correlation spectroscopy. The total drug content of NRG-SLNs was measured by high-performance liquid chromatography, and the encapsulation efficiency (EE) was determined by Sephadex gel-50 chromatography and high-performance liquid chromatography. The in vitro NRG release studies were carried out using a dialysis bag. The best cryoprotectant to prepare NRG-SLN lyophilized powder for future structural characterization was selected using differential scanning calorimetry, powder X-ray diffraction, and Fourier transform infrared spectroscopy. The short-term stability, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, cellular uptake, and pharmacokinetics in rats were studied after pulmonary administration of NRG-SLN lyophilized powder. Glycerol monostearate was selected to prepare SLNs, and the optimal formulation of NRG-SLNs was spherical in shape, with a particle size of 98 nm, a polydispersity index of 0.258, a zeta potential of -31.4 mV, a total drug content of 9.76 mg, an EE of 79.11%, and a cumulative drug release of 80% in 48 hours with a sustained profile. In addition, 5% mannitol (w

  19. Naringenin-loaded solid lipid nanoparticles: preparation, controlled delivery, cellular uptake, and pulmonary pharmacokinetics

    PubMed Central

    Ji, Peng; Yu, Tong; Liu, Ying; Jiang, Jie; Xu, Jie; Zhao, Ying; Hao, Yanna; Qiu, Yang; Zhao, Wenming; Wu, Chao

    2016-01-01

    Naringenin (NRG), a flavonoid compound, had been reported to exhibit extensive pharmacological effects, but its water solubility and oral bioavailability are only~46±6 µg/mL and 5.8%, respectively. The purpose of this study is to design and develop NRG-loaded solid lipid nanoparticles (NRG-SLNs) to provide prolonged and sustained drug release, with improved stability, involving nontoxic nanocarriers, and increase the bioavailability by means of pulmonary administration. Initially, a group contribution method was used to screen the best solid lipid matrix for the preparation of SLNs. NRG-SLNs were prepared by an emulsification and low-temperature solidification method and optimized using an orthogonal experiment approach. The morphology was examined by transmission electron microscopy, and the particle size and zeta potential were determined by photon correlation spectroscopy. The total drug content of NRG-SLNs was measured by high-performance liquid chromatography, and the encapsulation efficiency (EE) was determined by Sephadex gel-50 chromatography and high-performance liquid chromatography. The in vitro NRG release studies were carried out using a dialysis bag. The best cryoprotectant to prepare NRG-SLN lyophilized powder for future structural characterization was selected using differential scanning calorimetry, powder X-ray diffraction, and Fourier transform infrared spectroscopy. The short-term stability, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, cellular uptake, and pharmacokinetics in rats were studied after pulmonary administration of NRG-SLN lyophilized powder. Glycerol monostearate was selected to prepare SLNs, and the optimal formulation of NRG-SLNs was spherical in shape, with a particle size of 98 nm, a polydispersity index of 0.258, a zeta potential of −31.4 mV, a total drug content of 9.76 mg, an EE of 79.11%, and a cumulative drug release of 80% in 48 hours with a sustained profile. In addition, 5% mannitol (w

  20. Tiotropium bromide inhalation powder: a review of its use in the management of chronic obstructive pulmonary disease.

    PubMed

    Keating, Gillian M

    2012-01-22

    The anticholinergic agent tiotropium bromide (Spiriva®) is a long-acting bronchodilator that is indicated for the treatment of chronic obstructive pulmonary disease (COPD). This article reviews the clinical efficacy and tolerability of tiotropium bromide inhalation powder, administered using the HandiHaler® device, in patients with COPD, as well as reviewing its pharmacological properties and the results of pharmacoeconomic analyses. Shorter-term placebo-controlled trials in patients with COPD demonstrated significantly higher trough forced expiratory volume in 1 second (FEV(1)) responses with tiotropium bromide than with placebo, confirming it has a duration of action of ≥24 hours and is suitable for once-daily administration. Lung function improved to a greater extent with tiotropium bromide than with ipratropium bromide or, in most instances, salmeterol. Indacaterol was shown to be noninferior to tiotropium bromide in terms of the trough FEV(1) response. The large, 4-year UPLIFT® trial did not show a significant reduction in the annual rate of decline in FEV(1) with tiotropium bromide versus placebo in patients with COPD, although subgroup analyses demonstrated a significantly lower rate of decline with tiotropium bromide than with placebo in some patient groups (e.g. patients with moderate COPD, patients aged ≥50 years, patients not receiving maintenance therapy at baseline). Tiotropium bromide prevented exacerbations in patients with COPD, with a significantly lower exacerbation rate and a significantly longer time to first exacerbation seen with tiotropium bromide than with placebo or salmeterol. Exacerbation rates did not significantly differ between patients receiving tiotropium bromide and those receiving salmeterol/fluticasone propionate. Tiotropium bromide also had beneficial effects on health-related quality of life (HR-QOL) and other endpoints, such as dyspnoea and rescue medication use. Combination therapy with tiotropium bromide plus

  1. Chlorine-36, bromide, and the origin of spring water

    USGS Publications Warehouse

    Davis, S.N.; Cecil, L.D.; Zreda, M.; Moysey, S.

    2001-01-01

    Natural ratios of chlorine-36 (36Cl) to stable chlorine (i.e., 36Cl/Cl ?? 10-15) vary in shallow groundwater of the United States from about 50 in coastal areas to about 1400 in the northern Rocky Mountains. Ratios lower than these indicate the presence of chloride (Cl-) that has been isolated from the atmosphere for hundreds of thousands of years, if not longer. Higher ratios, which can exceed 5000, usually originate from fallout from testing thermonuclear devices in the western Pacific in the 1950s. Natural mass ratios of chloride to bromide (Cl-/Br-) in precipitation vary in the United States from about 250 in coastal areas to about 50 in the north-central states. Lower ratios may suggest contamination from human sources. Higher ratios, which may exceed 2000, commonly reflect the dissolution of halite. Seawater has a Cl-/Br- ratio of 290. Both 36Cl and Cl-/Br- ratios have been measured in 21 samples of spring water collected from springs in 10 different states. Brackish water from Saratoga Springs area in New York has low values for both 36Cl and Cl-/Br- ratios. This indicates that a large component of the water has a very deep origin. Brackish water from Alexander Springs in Florida has a low 36Cl ratio but a high Cl-/Br- ratio similar to seawater. This suggests the addition of ancient seawater that may be trapped in the aquifer. Big Spring in Iowa discharges water with a very high Cl-/Br- ratio but a moderate 36Cl ratio. The high ratio of Cl-/Br- may be produced by dissolution of road salt or agricultural chemicals. Of the 21 springs sampled, only 10 appeared to have potable water not significantly affected by human activity. Chlorine-36 from testing of nuclear devices is still being flushed out of four of the spring systems that were sampled. Thus, more than 45 years have passed since 36Cl was introduced into the aquifers feeding the springs and the systems, as yet, have not been purged. Published by Elsevier Science B.V.

  2. Bromine and bromide content in soils: Analytical approach from total reflection X-ray fluorescence spectrometry.

    PubMed

    Gallardo, Helena; Queralt, Ignasi; Tapias, Josefina; Candela, Lucila; Margui, Eva

    2016-08-01

    Monitoring total bromine and bromide concentrations in soils is significant in many environmental studies. Thus fast analytical methodologies that entail simple sample preparation and low-cost analyses are desired. In the present work, the possibilities and drawbacks of low-power total reflection X-ray fluorescence spectrometry (TXRF) for the determination of total bromine and bromide contents in soils were evaluated. The direct analysis of a solid suspension using 20 mg of fine ground soil (<63 μm) gave a 3.7 mg kg(-1) limit of detection for bromine which, in most cases, was suitable for monitoring total bromine content in soils (Br content range in soils = 5-40 mg kg(-1)). Information about bromide determination in soils is also possible by analyzing the Br content in water soil extracts. In this case, the TXRF analysis can be directly performed by depositing 10 μL of the internal standardized soil extract sample on a quartz glass reflector in a measuring time of 1500 s. The bromide limit of detection by this approach was 10 μg L(-1). Good agreement was obtained between the TXRF results for the total bromine and bromide determinations in soils and those obtained by other popular analytical techniques, e.g. energy dispersive X-ray fluorescence spectrometry (total bromine) and ionic chromatography (bromide). As a study case, the TXRF method was applied to study bromine accumulation in two agricultural soils fumigated with a methyl bromide pesticide and irrigated with regenerated waste water.

  3. Characterization of soluble bromide measurements and a case study of BrO observations during ARCTAS

    NASA Astrophysics Data System (ADS)

    Liao, J.; Huey, L. G.; Scheuer, E.; Dibb, J. E.; Stickel, R. E.; Tanner, D. J.; Neuman, J. A.; Nowak, J. B.; Choi, S.; Wang, Y.; Salawitch, R. J.; Canty, T.; Chance, K.; Kurosu, T.; Suleiman, R.; Weinheimer, A. J.; Shetter, R. E.; Fried, A.; Brune, W.; Anderson, B.; Zhang, X.; Chen, G.; Crawford, J.; Hecobian, A.; Ingall, E. D.

    2012-02-01

    A focus of the Arctic Research of the Composition of the Troposphere from Aircraft and Satellites (ARCTAS) mission was examination of bromine photochemistry in the spring time high latitude troposphere based on aircraft and satellite measurements of bromine oxide (BrO) and related species. The NASA DC-8 aircraft utilized a chemical ionization mass spectrometer (CIMS) to measure BrO and a mist chamber (MC) to measure soluble bromide. We have determined that the MC detection efficiency to molecular bromine (Br2), hypobromous acid (HOBr), bromine oxide (BrO), and hydrogen bromide (HBr) as soluble bromide (Br-) was 0.9±0.1, 1.06+0.30/-0.35, 0.4±0.1, and 0.95±0.1, respectively. These efficiency factors were used to estimate soluble bromide levels along the DC-8 flight track of 17 April 2008 from photochemical calculations constrained to in situ BrO measured by CIMS. During this flight, the highest levels of soluble bromide and BrO were observed and atmospheric conditions were ideal for the space-borne observation of BrO. The good agreement (R2 = 0.76; slope = 0.95; intercept = -3.4 pmol mol-1) between modeled and observed soluble bromide, when BrO was above detection limit (>2 pmol mol-1) under unpolluted conditions (NO<10 pmol mol-1), indicates that the CIMS BrO measurements were consistent with the MC soluble bromide and that a well characterized MC can be used to derive mixing ratios of some reactive bromine compounds. Tropospheric BrO vertical column densities (BrOVCD) derived from CIMS BrO observations compare well with BrOTROPVCD from OMI on 17 April 2008.

  4. Characterization of soluble bromide measurements and a case study of BrO observations during ARCTAS

    NASA Astrophysics Data System (ADS)

    Liao, J.; Huey, L. G.; Scheuer, E.; Dibb, J. E.; Stickel, R. E.; Tanner, D. J.; Neuman, J. A.; Nowak, J. B.; Choi, S.; Wang, Y.; Salawitch, R. J.; Canty, T.; Chance, K.; Kurosu, T.; Suleiman, R.; Weinheimer, A. J.; Shetter, R. E.; Fried, A.; Brune, W.; Anderson, B.; Zhang, X.; Chen, G.; Crawford, J.; Hecobian, A.; Ingall, E. D.

    2011-09-01

    A focus of the Arctic Research of the Composition of the Troposphere from Aircraft and Satellites (ARCTAS) mission was examination of bromine photochemistry in the spring time, high latitude troposphere based on aircraft and satellite measurements of BrO and related species. The NASA DC-8 aircraft utilized a chemical ionization mass spectrometry (CIMS) instrument to measure BrO and a mist chamber (MC) to measure soluble bromide. We have determined that the MC detection efficiency to molecular bromine (Br2), hypobromous acid (HOBr), bromine oxide (BrO), and hydrogen bromide (HBr) as soluble bromide (Br-) was 0.9±0.1, 1.06±0.30, 0.4±0.1, and 0.95±0.1, respectively. These efficiency factors were used to estimate soluble bromide levels along the DC-8 flight track of 17 April 2008 from photochemical calculations constrained to in situ BrO measured by CIMS. During this flight, the highest levels of soluble bromide and BrO were observed and atmospheric conditions were ideal for the space-borne observation of BrO. The good agreement (R2 = 0.76; slope = 0.98; intercept = -3.5 pptv) between modeled and observed soluble bromide, when BrO was above detection limit (>2 pptv) under unpolluted conditions (NO < 100 pptv), indicates that the CIMS BrO measurements were consistent with the MC soluble bromide. Tropospheric BrO vertical column densities (BROTROPVCD) derived from CIMS BrO observations compare well with BROTROPVCD from OMI on 17 April 2008.

  5. Comparative evaluation of the antiproliferative effect of cyclosporin A and gamma-interferon on normal and HPV-transformed keratinocytes by cell counting, MTT assay and tritiated thymidine incorporation.

    PubMed

    Marionnet, A V; Lizard, G; Chardonnet, Y; Schmitt, D

    1997-02-01

    We compared three techniques, the MTT tetrazolium assay, cell counting, and tritiated thymidine ([3H]TdR) incorporation assay to measure the antiproliferative effect of cyclosporin A (CsA) and interferon-gamma (IFN-gamma) on normal human skin keratinocyte cultures (NHK) used at the second passage and human papilomavirus type 16- and 18-transformed cell lines (EK16 and EK18) exposed continuously to the drugs for 3 days. The three techniques showed that under CsA (0.5 and 8 micrograms/ml) and IFN-gamma (5 and 160 U/ml) treatments the cells remained viable and that the growth of keratinocytes was inhibited. For IFN-gamma, the MTT colorimetric assay consistently underestimated its growth inhibitory activity as compared to cell counting or [3H]TdR incorporation, whatever the cells used. For high doses of CsA, MTT and cell counting gave similar percentages, of inhibitory activity whatever the cells; MTT underestimated this activity as compared to [3H]TdR incorporation only in NHK and EK18 cells, whereas similar results were obtained with EK16 cells. In conclusion, this investigations shows that MTT sensitivity differed with the drug and also according to the keratinocyte cultures. The MTT test is clearly not appropriate for study of IFN-gamma treatment whatever the keratinocytes used. Such discrepancies indicate that the MTT test should be done with care on cultures to measure the effects of drugs on cell growth; the growth inhibition should be carefully considered and it would be best if two different methods were used.

  6. Increased cytotoxicity of food-borne mycotoxins toward human cell lines in vitro via enhanced cytochrome p450 expression using the MTT bioassay.

    PubMed

    Lewis, C W; Smith, J E; Anderson, J G; Freshney, R I

    1999-11-01

    Eight food-borne mycotoxins epidemiologically implicated in human disease were tested for their cytotoxic effects on human cells previously immortalised and transfected to introduce human cytochrome p450 (CYP 450) genes. Such cells retain many characteristics of normal cell growth and differentiation while simultaneously having the potential of either increasing or decreasing the metabolic activity (cytotoxicity) of the challenging mycotoxins. The MTT assay provided an indication of cytotoxicity. Of the nine CYP450s introduced CYP1A2 was most effective, rendering the cells 540 times more sensitive than the control cells to aflatoxin B1, 28 times more sensitive to aflatoxin G1 and 8-fold more sensitive to ochratoxin A. CYP3A4 resulted in the cells being 211 times more toxic to aflatoxin B1 and 8-fold more toxic to aflatoxin G1 while CYP 2A6, CYP 3A5 and CYP 2E1 also produced observable effects. No increase in metabolic activity was found using cyclopiazonic acid, deoxynivalenol, fumonisin B1, patulin or T-2 toxin. CD5Os were calculated for the mycotoxins against the non-CYP-introduced control cells. There was almost a five order of magnitude difference between the most toxic, T-2 toxin (CD50 0.0057 microgram/ml) and the least toxic, fumonisin B1 (CD50 476.2 micrograms/ml). In vitro biological assays thus provide an excellent system for quantifying the often low CD50s expressed by mycotoxins in foods.

  7. Expanding the scope of PNA-encoded synthesis (PES): Mtt-protected PNA fully orthogonal to fmoc chemistry and a broad array of robust diversity-generating reactions.

    PubMed

    Chouikhi, Dalila; Ciobanu, Mihai; Zambaldo, Claudio; Duplan, Vincent; Barluenga, Sofia; Winssinger, Nicolas

    2012-10-01

    Nucleic acid-encoded libraries are emerging as an attractive and highly miniaturized format for the rapid identification of protein ligands. An important criterion in the synthesis of nucleic acid encoded libraries is the scope of reactions that can be used to introduce molecular diversity and devise divergent pathways for diversity-oriented synthesis (DOS). To date, the protecting group strategies that have been used in peptide nucleic acid (PNA) encoded synthesis (PES) have limited the choice of reactions used in the library synthesis to just a few prototypes. Herein, we describe the preparation of PNA monomers with a protecting group combination (Mtt/Boc) that is orthogonal to Fmoc-based synthesis and compatible with a large palette of reactions that have been productively used in DOS (palladium cross-couplings, metathesis, reductive amination, amidation, heterocycle formation, nucleophilic addition, conjugate additions, Pictet-Spengler cyclization). We incorporate γ-modifications in the PNA backbone that are known to enhance hybridization and solubility. We demonstrate the robustness of this strategy with a library synthesis that is characterized by MALDI MS analysis at every step.

  8. A novel explicit approach to model bromide and pesticide transport in soils containing macropores

    NASA Astrophysics Data System (ADS)

    Klaus, J.; Zehe, E.

    2011-01-01

    The present study tests whether an explicit treatment of worm burrows is feasible for simulating water flow, bromide and pesticide transport in structured heterogeneous soils. The essence is to represent worm burrows as morphologically connected paths of low flow resistance in the spatially highly resolved model domain. A recent Monte Carlo study (Klaus and Zehe, 2010) revealed that this approach allowed successful reproduction of tile drain event discharge recorded during an irrigation experiment at a tile drained field site. However, several "hillslope architectures" that were all consistent with the available extensive data base allowed a good reproduction of tile drain flow response. Our second objective was thus to find out whether this "equifinality" in spatial model setups may be reduced when including bromide tracer data in the model falsification process. We thus simulated transport of bromide and Isoproturon (IPU) for the 13 spatial model setups, which performed best with respect to reproduce tile drain event discharge, without any further calibration. All model setups allowed a very good prediction of the temporal dynamics of cumulated bromide leaching into the tile drain, while only four of them matched the accumulated water balance and accumulated bromide loss into the tile drain. The number of behavioural model architectures could thus be reduced to four. One of those setups was used for simulating transport of IPU, using different parameter combinations to characterise adsorption according to the Footprint data base. Simulations could, however, only reproduce the observed leaching behaviour, when we allowed for retardation coefficients that were very close to one.

  9. Utility of bromide and heat tracers for aquifer characterization affected by highly transient flow conditions

    NASA Astrophysics Data System (ADS)

    Ma, Rui; Zheng, Chunmiao; Zachara, John M.; Tonkin, Matthew

    2012-08-01

    A tracer test using both bromide and heat tracers conducted at the Integrated Field Research Challenge site in Hanford 300 Area (300A), Washington, provided an instrument for evaluating the utility of bromide and heat tracers for aquifer characterization. The bromide tracer data were critical to improving the calibration of the flow model complicated by the highly dynamic nature of the flow field. However, most bromide concentrations were obtained from fully screened observation wells, lacking depth-specific resolution for vertical characterization. On the other hand, depth-specific temperature data were relatively simple and inexpensive to acquire. However, temperature-driven fluid density effects influenced heat plume movement. Moreover, the temperature data contained "noise" caused by heating during fluid injection and sampling events. Using the hydraulic conductivity distribution obtained from the calibration of the bromide transport model, the temperature depth profiles and arrival times of temperature peaks simulated by the heat transport model were in reasonable agreement with observations. This suggested that heat can be used as a cost-effective proxy for solute tracers for calibration of the hydraulic conductivity distribution, especially in the vertical direction. However, a heat tracer test must be carefully designed and executed to minimize fluid density effects and sources of noise in temperature data. A sensitivity analysis also revealed that heat transport was most sensitive to hydraulic conductivity and porosity, less sensitive to thermal distribution factor, and least sensitive to thermal dispersion and heat conduction. This indicated that the hydraulic conductivity remains the primary calibration parameter for heat transport.

  10. Mepenzolate bromide promotes diabetic wound healing by modulating inflammation and oxidative stress

    PubMed Central

    Zheng, Yongjun; Wang, Xingtong; Ji, Shizhao; Tian, Song; Wu, Haibin; Luo, Pengfei; Fang, He; Wang, Li; Wu, Guosheng; Xiao, Shichu; Xia, Zhaofan

    2016-01-01

    Diabetic wounds are characterized by persistent inflammation and the excessive production of reactive oxygen species, thus resulting in impaired wound healing. Mepenzolate bromide, which was originally used to treat gastrointestinal disorders in clinical settings, has recently been shown to display beneficial effects in chronic obstructive pulmonary disease and pulmonary fibrosis of a mouse model by inhibiting inflammatory responses and reducing oxidative stress. However,the role of mepenzolate bromide in diabetic wound healing is still unclear. In this study, full-thickness excisional skin wounds were created on the backs of db/db mice, and mepenzolate bromide was topically applied to the wound bed. We found that mepenzolate bromide significantly promoted diabetic wound healing by measuring wound closure rate and histomorphometric analyses. Further studies showed that inflammation was inhibited by assessing the number of macrophages and levels of pro-inflammatory cytokines and pro-healing cytokines in the wounds. Furthermore, oxidative stress was reduced by monitoring the levels of MDA and H2O2 and the activities of glutathione peroxidase and catalase in the wounds. These results demonstrated the potential application of mepenzolate bromide for treating diabetic ulcers and other chronic wounds in clinics. PMID:27398156

  11. [Successful lobectomy in 3 lung cancer cases with severe COPD after treatment with tiotropium bromide].

    PubMed

    Matsuyama, Wataru; Suetsugu, Takayuki; Kawabata, Takashi; Kubota, Shingo; Iwakawa, Jun; Higashimoto, Ikkou; Osame, Mitsuhiro; Arimura, Kimiyoshi

    2007-02-01

    It is well known that lung cancer patients with severe chronic obstructive pulmonary disease (COPD) have a higher risk of postoperative complications than patients without COPD. However, the information regarding preoperative treatment to improve pulmonary function of the lung cancer patients with severe COPD is limited. Here, we report 3 lung cancer cases with severe COPD. Although all patients received medication without tiotropium bromide in combination with pulmonary rehabilitation for 1 or 2 months, their pulmonary function did not improve and the predicted postoperative FEV1/predicted FEV1 was below 40% in all cases. After the approval in Japan for use of tiotropium bromide in the treatment of COPD, all patients were treated with tiotropium bromide. The pulmonary function in all patients improved 2-4 weeks after the start of tiotropium bromide, and we performed lobectomy safely. Currently all patients maintain good pulmonary function without recurrence of lung cancer. We propose that treatment of tiotropium bromide might be one of the effective preoperative methods to improve pulmonary function of lung cancer patients with severe COPD.

  12. Rocuronium Bromide Inhibits Inflammation and Pain by Suppressing Nitric Oxide Production and Enhancing Prostaglandin E2 Synthesis in Endothelial Cells

    PubMed Central

    2016-01-01

    Purpose Rocuronium bromide is a nondepolarizing neuromuscular blocking drug and has been used as an adjunct for relaxation or paralysis of the skeletal muscles, facilitation of endotracheal intubation, and improving surgical conditions during general anesthesia. However, intravenous injection of rocuronium bromide induces injection pain or withdrawal movement. The exact mechanism of rocuronium bromide-induced injection pain or withdrawal movement is not yet understood. We investigated whether rocuronium bromide treatment is involved in the induction of inflammation and pain in vascular endothelial cells. Methods For this study, calf pulmonary artery endothelial (CPAE) cells were used, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Western blot, nitric oxide detection, and prostaglandin E2 immunoassay were conducted. Results Rocuronium bromide treatment inhibited endothelial nitric oxide synthase and suppressed nitric oxide production in CPAE cells. Rocuronium bromide activated cyclooxygenase-2, inducible nitric oxide synthase and increased prostaglandin E2 synthesis in CPAE cells. Conclusions Rocuronium bromide induced inflammation and pain in CPAE cells. Suppressing nitric oxide production and enhancing prostaglandin E2 synthesis might be associated with rocuronium bromide-induced injection pain or withdrawal movement. PMID:28043117

  13. Methyl bromide: ocean sources, ocean sinks, and climate sensitivity.

    PubMed

    Anbar, A D; Yung, Y L; Chavez, F P

    1996-03-01

    The oceans play an important role in the geochemical cycle of methyl bromide (CH3Br), the major carrier of O3-destroying bromine to the stratosphere. The quantity of CH3Br produced annually in seawater is comparable to the amount entering the atmosphere each year from natural and anthropogenic sources. The production mechanism is unknown but may be biological. Most of this CH3Br is consumed in situ by hydrolysis or reaction with chloride. The size of the fraction which escapes to the atmosphere is poorly constrained; measurements in seawater and the atmosphere have been used to justify both a large oceanic CH3Br flux to the atmosphere and a small net ocean sink. Since the consumption reactions are extremely temperature-sensitive, small temperature variations have large effects on the CH3Br concentration in seawater, and therefore on the exchange between the atmosphere and the ocean. The net CH3Br flux is also sensitive to variations in the rate of CH3Br production. We have quantified these effects using a simple steady state mass balance model. When CH3Br production rates are linearly scaled with seawater chlorophyll content, this model reproduces the latitudinal variations in marine CH3Br concentrations observed in the east Pacific Ocean by Singh et al. [1983] and by Lobert et al. [1995]. The apparent correlation of CH3Br production with primary production explains the discrepancies between the two observational studies, strengthening recent suggestions that the open ocean is a small net sink for atmospheric CH3Br, rather than a large net source. The Southern Ocean is implicated as a possible large net source of CH3Br to the atmosphere. Since our model indicates that both the direction and magnitude of CH3Br exchange between the atmosphere and ocean are extremely sensitive to temperature and marine productivity, and since the rate of CH3Br production in the oceans is comparable to the rate at which this compound is introduced to the atmosphere, even small

  14. Methyl bromide: ocean sources, ocean sinks, and climate sensitivity

    NASA Technical Reports Server (NTRS)

    Anbar, A. D.; Yung, Y. L.; Chavez, F. P.

    1996-01-01

    The oceans play an important role in the geochemical cycle of methyl bromide (CH3Br), the major carrier of O3-destroying bromine to the stratosphere. The quantity of CH3Br produced annually in seawater is comparable to the amount entering the atmosphere each year from natural and anthropogenic sources. The production mechanism is unknown but may be biological. Most of this CH3Br is consumed in situ by hydrolysis or reaction with chloride. The size of the fraction which escapes to the atmosphere is poorly constrained; measurements in seawater and the atmosphere have been used to justify both a large oceanic CH3Br flux to the atmosphere and a small net ocean sink. Since the consumption reactions are extremely temperature-sensitive, small temperature variations have large effects on the CH3Br concentration in seawater, and therefore on the exchange between the atmosphere and the ocean. The net CH3Br flux is also sensitive to variations in the rate of CH3Br production. We have quantified these effects using a simple steady state mass balance model. When CH3Br production rates are linearly scaled with seawater chlorophyll content, this model reproduces the latitudinal variations in marine CH3Br concentrations observed in the east Pacific Ocean by Singh et al. [1983] and by Lobert et al. [1995]. The apparent correlation of CH3Br production with primary production explains the discrepancies between the two observational studies, strengthening recent suggestions that the open ocean is a small net sink for atmospheric CH3Br, rather than a large net source. The Southern Ocean is implicated as a possible large net source of CH3Br to the atmosphere. Since our model indicates that both the direction and magnitude of CH3Br exchange between the atmosphere and ocean are extremely sensitive to temperature and marine productivity, and since the rate of CH3Br production in the oceans is comparable to the rate at which this compound is introduced to the atmosphere, even small

  15. Current and Potential Future Bromide Loads from Coal-Fired Power Plants in the Allegheny River Basin and Their Effects on Downstream Concentrations.

    PubMed

    Good, Kelly D; VanBriesen, Jeanne M

    2016-09-06

    The presence of bromide in rivers does not affect ecosystems or present a human health risk; however, elevated concentrations of bromide in drinking water sources can lead to difficulty meeting drinking water disinfection byproduct (DBP) regulations. Recent attention has focused on oil and gas wastewater and coal-fired power plant wet flue gas desulfurization (FGD) wastewater bromide discharges. Bromide can be added to coal to enhance mercury removal, and increased use of bromide at some power plants is expected. Evaluation of potential increases in bromide concentrations from bromide addition for mercury control is lacking. The present work utilizes bromide monitoring data in the Allegheny River and a mass-balance approach to elucidate bromide contributions from anthropogenic and natural sources under current and future scenarios. For the Allegheny River, the current bromide is associated approximately 49% with oil- and gas-produced water discharges and 33% with coal-fired power plants operating wet FGD, with 18% derived from natural sources during mean flow conditions in August. Median wet FGD bromide loads could increase 3-fold from 610 to 1900 kg/day if all plants implement bromide addition for mercury control. Median bromide concentrations in the lower Allegheny River in August would rise to 410, 200, and 180 μg/L under low-, mean-, and high-flow conditions, respectively, for the bromide-addition scenario.

  16. Kinetics adsorption study of the ethidium bromide by graphene oxide as adsorbent from aqueous matrices

    NASA Astrophysics Data System (ADS)

    Rajabi, M.; Moradi, O.; Zare, K.

    2017-01-01

    In this study of ethidium bromide, adsorption from aqueous matrices by graphene oxide as adsorbent was investigated. Influencing parameters in the adsorption study included contact time, temperature, and pH. The optimum time was selected 17 min, and the best value of pH was determined at 8. All adsorption experiments were performed at 298 K temperature. The maximum wavelength of ethidium bromide was 475 nm. The Elovich, four types of the pseudo-second-order, the pseudo-first-order, and intra-particle diffusion kinetic adsorption models were used for kinetic study, and the results show that adsorption of ethidium bromide on graphene oxide surface best complied with type (I) of the pseudo-second-order kinetic model.

  17. A comparison between the gastric and salivary concentration of iodide, pertechnetate, and bromide in man

    PubMed Central

    Harden, R. McG.; Alexander, W. D.; Shimmins, J.; Chisholm, D.

    1969-01-01

    The concentration of iodide (I−) and pertechnetate (TcO4−) and bromide (Br−) has been measured simultaneously in gastric juice and parotid saliva. The combined gastric and salivary clearance for iodide and pertechnetate is more than twice the clearance of these ions by the thyroid gland. The concentration of the ions was in the order I−>TcO4−>Br− in both gastric juice and saliva. Differences exist between the secretion of iodide, pertechnetate, and bromide. Bromide, in contrast to iodide and pertechnetate, was found to be more concentrated in gastric juice than in saliva. The ratio of the iodide to pertechnetate clearance was greater in gastric juice than in saliva. PMID:5358585

  18. Characterization of micelle formation of dodecyldimethyl-N-2-phenoxyethylammonium bromide in aqueous solution.

    PubMed

    Khatua, Dibyendu; Gupta, Abhisek; Dey, Joykrishna

    2006-06-01

    Aggregation behavior of dodecyldimethyl-N-2-phenoxyethylammonium bromide commonly called domiphen bromide (DB) was studied in aqueous solution. The Krafft temperature of the surfactant was measured. The surfactant has been shown to form micellar structures in a wide concentration range. The critical micelle concentration was determined by surface tension, conductivity, and fluorescence methods. The conductivity data were also employed to determine the degree of surfactant counterion dissociation. The changes in Gibb's free energy, enthalpy, and entropy of the micellization process were determined at different temperature. The steady-state fluorescence quenching measurements with pyrene and N-phenyl-1-naphthylamine as fluorescence probes were performed to obtain micellar aggregation number. The results were compared with those of dodecyltrimethylammonium bromide (DTAB) surfactant. The micelle formation is energetically more favored in DB compared to that in DTAB. The 1H-NMR spectra were used to show that the 2-phenoxyethyl group, which folds back onto the micellar surface facilitates aggregate formation in DB.

  19. Seeded Growth of Monodisperse Gold Nanorods Using Bromide-Free Surfactant Mixtures

    SciTech Connect

    Ye, XC; Gao, YZ; Chen, J; Reifsnyder, DC; Zheng, C; Murray, CB

    2013-05-01

    We demonstrate for the first time that monodisperse gold nanorods (NRs) with broadly tunable dimensions and longitudinal surface plasmon resonances can be synthesized using a bromide-free surfactant mixture composed of alkyltrimethylammonium chloride and sodium oleate. It is found that uniform gold NRs can be obtained even with an iodide concentration approaching 100 mu M in the growth solution. In contrast to conventional wisdom, our results provide conclusive evidence that neither bromide as the surfactant counterion nor a high concentration of bromide ions in the growth solution is essential for gold NR formation. Correlated electron microscopy study of three-dimensional structures of gold NRs reveals a previously unprecedented octagonal prismatic structure enclosed predominantly by high index {310} crystal planes. These findings should have profound implications for a comprehensive mechanistic understanding of seeded growth of anisotropic metal nanocrystals.

  20. [Extraction-spectrophotometric determination of carbethopendecinium-bromide in drug preparations].

    PubMed

    Lasková, M; Blesová, M

    2003-09-01

    The paper elaborated the conditions for the determination of carbethopendecinium bromide with bromthymol blue and methyl orange (concentration of the drug and the colouring agent, pH of the aqueous phase, period of extraction with chloroform). Within a carbethopendecinium bromide concentration range of 1.10(-5) to 6.10(-5) mol/l, the linear dependence of absorbance of the ion-associate with the colouring agent on concentration was confirmed. At pH 3 and pH 10, the above-described procedure was used to determine the content of carbethopendecinium bromide in medicinal preparations (eye and nasal drops) using the calibration curve and comparison with one standard. The results were reproducible and the found deviations from the declared content ranged within a tolerance of +/- 10%.

  1. Salty glycerol versus salty water surface organization: bromide and iodide surface propensities.

    PubMed

    Huang, Zishuai; Hua, Wei; Verreault, Dominique; Allen, Heather C

    2013-07-25

    Salty NaBr and NaI glycerol solution interfaces are examined in the OH stretching region using broadband vibrational sum frequency generation (VSFG) spectroscopy. Raman and infrared (IR) spectroscopy are used to further understand the VSFG spectroscopic signature. The VSFG spectra of salty glycerol solutions reveal that bromide and iodide anions perturb the interfacial glycerol organization in a manner similar as that found in aqueous halide salt solutions, thus confirming the presence of bromide and iodide anions at the glycerol surface. Surface tension measurements are consistent with the surface propensity suggested by the VSFG data and also show that the surface excess increases with increasing salt concentration, similar to that of water. In addition, iodide is shown to have more surface prevalence than bromide, as has also been determined from aqueous solutions. These results suggest that glycerol behaves similarly to water with respect to surface activity and solvation of halide anions at its air/liquid interface.

  2. Cytotoxicity of Agaricus sylvaticus in non-tumor cells (NIH/3T3) and tumor (OSCC-3) using tetrazolium (MTT) assay.

    PubMed

    Orsine, Joice Vinhal Costa; Marques Brito, Luíssa; Silva, Renata Carvalho; Santos Almeida, Maria de Fátima Menezes; Novaes, Maria Rita Carvalho Garbi

    2013-01-01

    The purpose of this study was to assess the cytotoxic effect of the non-fractionated aqueous extract of A. sylvaticus mushroom in cultures of non-tumor cells (NIH3T3) and tumor cells (OSCC-3). The cells were maintained in DMEN cell culture medium added of 10% of fetal bovine serum and 1% antibiotic. For the cytotoxicity test we prepared the aqueous mushroom extract at concentrations of 0.01 mg.ml⁻¹, 0.02 mg.ml⁻¹, 0.04 mg.ml⁻¹, 0.08 mg.ml⁻¹, 0.16 mg.ml⁻¹, and 0.32 mg.ml⁻¹. For the culture, 2 x 10⁵ cells/ml was deposited in 96-well microplates during 24 hour incubation with subsequent exchange of medium by another containing the mushroom concentrations. After 24 hour incubation the medium was discarded and 100 ml of tetrazolium blue (MTT) was added at a concentration of 5 mg.ml⁻¹. The microplates were incubated for 2 h at 37° C. Spectrophotometric analysis was performed using 570 nm wavelength. From the values of the optical densities we determined the drug concentration capable of reducing cell viability by 50%. Therefore, the mushroom A. sylvaticus, at all concentrations tested, did not show cytotoxic effects, once the inhibitory concentration (IC₅₀) obtained for tumor cells OSCC-3 was 0.06194 mg.ml⁻¹, and the IC₅₀ checked for non-tumor cells NIH3T3 was 0,06468 mg.ml⁻¹. This test made it possible to determine that A. sylvaticus mushroom has no cytotoxic effects, suggesting its use safe for human consumption.

  3. In-Depth Assessment of the Palladium-Catalyzed Fluorination of Five-Membered Heteroaryl Bromides.

    PubMed

    Milner, Phillip J; Yang, Yang; Buchwald, Stephen L

    2015-10-12

    A thorough investigation of the challenging Pd-catalyzed fluorination of five-membered heteroaryl bromides is presented. Crystallographic studies and density functional theory (DFT) calculations suggest that the challenging step of this transformation is C-F reductive elimination of five-membered heteroaryl fluorides from Pd(II) complexes. On the basis of these studies, we have found that various heteroaryl bromides bearing phenyl groups in the ortho position can be effectively fluorinated under catalytic conditions. Highly activated 2-bromoazoles, such as 8-bromocaffeine, are also viable substrates for this reaction.

  4. Lithium bromide-induced structural changes in a nickel bis-alkoxide complex.

    PubMed

    Ichiokai, Hiromi; Vicic, David A

    2013-01-01

    The bis-alkoxide [(DEAMP)2Ni] (1, DEAMP = 1-(diethylamino)-2-methylpropan-2-olate) was found to react with trace amounts of lithium bromide to afford the bis-LiBr adduct 2, in which the oxygens of the DEAMP ligand coordinate to lithium to form a chiral-at-metal complex. This new complex is five-coordinate at nickel, and contains nickel and oxygen atoms which are all chiral. One diastereomer precipitates from pentane solution. The two lithium ions rigidify the new structure in the solid state by coordinating to the oxygen and bromide atoms.

  5. One-pot synthesis of 1-iodoalkynes and trisubstituted alkenes from benzylic and allylic bromides.

    PubMed

    Pelletier, Guillaume; Lie, Sharon; Mousseau, James J; Charette, André B

    2012-11-02

    1-Iodoalkynes are formed in moderate to high yields from readily accessible benzylic and allylic alkyl bromides by a one-pot homologation/double elimination procedure with iodoform (CHI(3)). The developed conditions include facile purification and avoid the use of an excess of triphenylphosphine (PPh(3)), as described in classical Corey-Fuchs iodoalkynylation conditions. Replacing CHI(3) with CHI(2)Cl allows the isolation of the corresponding gem-(Z)-chloro-(E)-iodoalkene in good yield and stereoselectivity. Moreover, the use of benzhydryl bromides as nucleophiles enables the synthesis of trisubstituted alkenes under similar reaction conditions.

  6. Improved light olefin yield from methyl bromide coupling over modified SAPO-34 molecular sieves.

    PubMed

    Zhang, Aihua; Sun, Shouli; Komon, Zachary J A; Osterwalder, Neil; Gadewar, Sagar; Stoimenov, Peter; Auerbach, Daniel J; Stucky, Galen D; McFarland, Eric W

    2011-02-21

    As an alternative to the partial oxidation of methane to synthesis gas followed by methanol synthesis and the subsequent generation of olefins, we have studied the production of light olefins (ethylene and propylene) from the reaction of methyl bromide over various modified microporous silico-aluminophosphate molecular-sieve catalysts with an emphasis on SAPO-34. Some comparisons of methyl halides and methanol as reaction intermediates in their conversion to olefins are presented. Increasing the ratio of Si/Al and incorporation of Co into the catalyst framework improved the methyl bromide yield of light olefins over that obtained using standard SAPO-34.

  7. Bromidotetra-kis-(2-ethyl-1H-imidazole-κN (3))copper(II) bromide.

    PubMed

    Godlewska, Sylwia; Kelm, Harald; Krüger, Hans-Jörg; Dołęga, Anna

    2012-12-01

    The Cu(II) ion in the title mol-ecular salt, [CuBr(C5H8N2)4]Br, is coordinated in a square-pyramidal geometry by four N atoms of imidazole ligands and one bromide anion in the apical position. In the crystal, the ions are linked by N-H⋯Br hydrogen bonds involving both the coordinating and the free bromide species as acceptors. A C-H⋯Br inter-action is also observed. Overall, a three-dimensional network results.

  8. Breakdown of ionic character of molecular alkali bromides in inner-valence photoionization

    SciTech Connect

    Karpenko, A. Iablonskyi, D.; Kettunen, J. A.; Cao, W.; Huttula, M.; Aksela, H.; Urpelainen, S.

    2014-05-28

    The inner-valence region of alkali bromide XBr (X=Li, Na, K, Rb) vapours has been studied experimentally by means of synchrotron radiation excited photoelectron spectroscopy. Experimental spectra were analyzed by comparing them with available theoretical results and previous experiments. Ionic character of alkali bromides is seen to change in the inner-valence region with increasing atomic number of the alkali atom. A mechanism involving mixing between Br 4s and Rb 4p orbitals has been suggested to account for the fine structure observed in inner-valence ionization region of RbBr.

  9. Bromide, Chloride, and Sulfate Concentrations, and Specific Conductance, Lake Texoma, Texas and Oklahoma, 2007-08

    USGS Publications Warehouse

    Baldys, Stanley

    2009-01-01

    The U.S. Geological Survey, in cooperation with the City of Dallas Water Utilities Division, collected water-quality data from 11 sites on Lake Texoma, a reservoir on the Texas-Oklahoma border, during April 2007-September 2008. At 10 of the sites, physical properties (depth, specific conductance, pH, temperature, dissolved oxygen, and alkalinity) were measured and samples were collected for analysis of selected dissolved constituents (bromide, calcium, magnesium, potassium, sodium, carbonate, bicarbonate, chloride, and sulfate); at one site, only physical properties were measured. The primary constituent of interest was bromide. Bromate can form when ozone is used to disinfect raw water containing bromide, and bromate is a suspected human carcinogen. Chloride and sulfate were of secondary interest. Only the analytical results for bromide, chloride, sulfate, and measured specific conductance are discussed in this report. Median dissolved bromide concentrations ranged from 0.28 to 0.60 milligrams per liter. The largest median dissolved bromide concentration (0.60 milligram per liter at site 11) was from the Red River arm of Lake Texoma. Dissolved bromide concentrations generally were larger in the Red River arm of Lake Texoma than in the Washita arm of the lake. Median dissolved chloride concentrations were largest in the Red River arm of Lake Texoma at site 11 (431 milligrams per liter) and smallest at site 8 (122 milligrams per liter) in the Washita arm. At site 11 in the Red River arm, the mean and median chloride concentrations exceeded the secondary maximum contaminant level of 300 milligrams per liter for chloride established by the 'Texas Surface Water Quality Standards' for surface-water bodies designated for the public water supply use. Median dissolved sulfate concentrations ranged from 182 milligrams per liter at site 4 in the Big Mineral arm to 246 milligrams per liter at site 11 in the Red River arm. None of the mean or median sulfate concentrations

  10. In-Depth Assessment of the Palladium-Catalyzed Fluorination of Five-Membered Heteroaryl Bromides

    PubMed Central

    2015-01-01

    A thorough investigation of the challenging Pd-catalyzed fluorination of five-membered heteroaryl bromides is presented. Crystallographic studies and density functional theory (DFT) calculations suggest that the challenging step of this transformation is C–F reductive elimination of five-membered heteroaryl fluorides from Pd(II) complexes. On the basis of these studies, we have found that various heteroaryl bromides bearing phenyl groups in the ortho position can be effectively fluorinated under catalytic conditions. Highly activated 2-bromoazoles, such as 8-bromocaffeine, are also viable substrates for this reaction. PMID:27056379

  11. Improved synthesis of aryltriethoxysilanes via palladium(0)-catalyzed silylation of aryl iodides and bromides with triethoxysilane.

    PubMed

    Manoso, A S; DeShong, P

    2001-11-02

    The scope of the palladium-catalyzed silylation of aryl halides with triethoxysilane has been expanded to include aryl bromides. A more general Pd(0) catalyst/ligand system has been developed that activates bromides and iodides: palladium(0) dibenzylideneacetone (Pd(dba)(2)) is activated with 2-(di-tert-butylphosphino)biphenyl (Buchwald's ligand) (1:2 mol ratio of Pd/phosphine). Electron-rich para- and meta-substituted aryl halides (including unprotected aniline and phenol derivatives) undergo silylation to form the corresponding aryltriethoxysilane in fair to excellent yield; however, ortho-substituted aryl halides failed to be silylated.

  12. Structure and dynamics of cetyltrimethylammonium bromide water-in-oil microemulsions

    SciTech Connect

    Lang, J.; Zana, R. ); Mascolo, G.; Luisi, P.L. )

    1990-04-05

    The state of water, the droplet size, and interdroplet exchange of reactants between colliding droplets have been investigated by NMR, time-resolved fluorescence quenching, and electrical conductivity in water-in-oil microemulsions made of cetyltrimethylammonium bromide (CTAB) in chloroform/isooctane (2/1, v/v). The effect of the two additives cetyl bromide (CB) and trimethylamine (TMA) on the properties of the droplets has also been investigated. No interdroplet exchange of reactants has been detected in the absence of additives.

  13. Active Control of Interface Shape During the Crystal Growth of Lead Bromide

    NASA Technical Reports Server (NTRS)

    Duval, W. M. B.; Batur, C.; Singh, N. B.

    2003-01-01

    A thermal model for predicting and designing the furnace temperature profile was developed and used for the crystal growth of lead bromide. The model gives the ampoule temperature as a function of the furnace temperature, thermal conductivity, heat transfer coefficients, and ampoule dimensions as variable parameters. Crystal interface curvature was derived from the model and it was compared with the predicted curvature for a particular furnace temperature and growth parameters. Large crystals of lead bromide were grown and it was observed that interface shape was in agreement with the shape predicted by this model.

  14. 40 CFR 180.123a - Inorganic bromide residues in peanut hay and peanut hulls; statement of policy.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Inorganic bromide residues in peanut hay and peanut hulls; statement of policy. 180.123a Section 180.123a Protection of Environment... RESIDUES IN FOOD Specific Tolerances § 180.123a Inorganic bromide residues in peanut hay and peanut...

  15. 40 CFR 180.123a - Inorganic bromide residues in peanut hay and peanut hulls; statement of policy.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 24 2014-07-01 2014-07-01 false Inorganic bromide residues in peanut hay and peanut hulls; statement of policy. 180.123a Section 180.123a Protection of Environment... RESIDUES IN FOOD Specific Tolerances § 180.123a Inorganic bromide residues in peanut hay and peanut...

  16. 40 CFR 180.123a - Inorganic bromide residues in peanut hay and peanut hulls; statement of policy.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 25 2013-07-01 2013-07-01 false Inorganic bromide residues in peanut hay and peanut hulls; statement of policy. 180.123a Section 180.123a Protection of Environment... RESIDUES IN FOOD Specific Tolerances § 180.123a Inorganic bromide residues in peanut hay and peanut...

  17. 40 CFR 180.123a - Inorganic bromide residues in peanut hay and peanut hulls; statement of policy.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 24 2011-07-01 2011-07-01 false Inorganic bromide residues in peanut hay and peanut hulls; statement of policy. 180.123a Section 180.123a Protection of Environment... RESIDUES IN FOOD Specific Tolerances § 180.123a Inorganic bromide residues in peanut hay and peanut...

  18. 40 CFR 180.123a - Inorganic bromide residues in peanut hay and peanut hulls; statement of policy.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 25 2012-07-01 2012-07-01 false Inorganic bromide residues in peanut hay and peanut hulls; statement of policy. 180.123a Section 180.123a Protection of Environment... RESIDUES IN FOOD Specific Tolerances § 180.123a Inorganic bromide residues in peanut hay and peanut...

  19. PKR promotes choroidal neovascularization via upregulating the PI3K/Akt signaling pathway in VEGF expression

    PubMed Central

    Zhu, Manhui; Liu, Xiaojuan; Wang, Shengcun; Miao, Jin; Wu, Liucheng; Yang, Xiaowei; Wang, Ying; Kang, Lihua; Li, Wendie; Cui, Chen; Sang, Aimin

    2016-01-01

    Purpose The aim of this study was to investigate the functions of dsRNA-activated protein kinase (PKR) in choroidal neovascularization (CNV) and related signaling pathways in the production of vascular endothelial growth factor (VEGF). Methods A chemical hypoxia model of in vitro RF/6A cells, a rhesus choroid-retinal endothelial cell line, was established by adding cobalt chloride (CoCl2) to the culture medium. PKR, phosphophosphatidylinositol 3-kinase (p-PI3K), phosphoprotein kinase B (p-Akt), and VEGF protein levels in RF/6A cells were detected with western blotting. PKR siRNA and the PI3K inhibitor LY294002 were used to evaluate the roles of the PKR and PI3K signaling pathways in VEGF expression with western blotting. In an ARPE-19 (RPE cell line) and RF/6A cell coculture system, proliferation, migration, and tube formation of RF/6A cells under hypoxic conditions were measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), Transwell, and Matrigel Transwell assays, respectively. In vivo CNV lesions were induced in C57BL/6J mice using laser photocoagulation. The mice were euthanized in a timely manner, and the eyecups were dissected from enucleated eyes. PKR, p-PI3K, p-Akt, and VEGF protein levels in tissues were detected with western blotting. To evaluate the leakage area, fundus fluorescein angiography and choroidal flat mount were performed on day 7 after intravitreal injection of an anti-PKR monoclonal antibody. Results The in vitro RF/6A cell chemical hypoxia model showed that PKR expression was upregulated in parallel with p-PI3K, p-Akt, and VEGF expression, peaking at 12 h. PKR siRNA downregulated PKR, p-PI3K, p-Akt, and VEGF expression. In addition, the PI3K inhibitor LY294002 greatly decreased the p-PI3K, p-Akt, and VEGF protein levels, but PKR expression was unaffected, indicating that Akt was a downstream molecule of PKR that upregulated VEGF expression. In the ARPE-19 (RPE cell line) and RF/6A cell coculture system, PKR si

  20. EPA Takes Action Against Illegal Methyl Bromide Use in Puerto Rico

    EPA Pesticide Factsheets

    (New York, N.Y.) As part of its ongoing work to address the illegal use of toxic pesticides containing methyl bromide in the Caribbean, the U.S. Environmental Protection Agency has issued legal complaints against two individuals and a pest control company

  1. Hydrology and Water Quality near Bromide Pavilion in Chickasaw National Recreation Area, Murray County, Oklahoma, 2000

    USGS Publications Warehouse

    Andrews, William J.; Burrough, Steven P.

    2002-01-01

    The Bromide Pavilion in Chickasaw National Recreation Area drew many thousands of people annually to drink the mineral-rich waters piped from nearby Bromide and Medicine Springs. Periodic detection of fecal coliform bacteria in water piped to the pavilion from the springs, low yields of the springs, or flooding by adjacent Rock Creek prompted National Park Service officials to discontinue piping of the springs to the pavilion in the 1970s. Park officials would like to resume piping mineralized spring water to the pavilion to restore it as a visitor attraction, but they are concerned about the ability of the springs to provide sufficient quantities of potable water. Pumping and sampling of Bromide and Medicine Springs and Rock Creek six times during 2000 indicate that these springs may not provide sufficient water for Bromide Pavilion to supply large numbers of visitors. A potential problem with piping water from Medicine Spring is the presence of an undercut, overhanging cliff composed of conglomerate, which may collapse. Evidence of intermittent inundation of the springs by Rock Creek and seepage of surface water into the spring vaults from the adjoining creek pose a threat of contamination of the springs. Escherichia coli, fecal coliform, and fecal streptococcal bacteria were detected in some samples from the springs, indicating possible fecal contamination. Cysts of Giardia lamblia and oocysts of Cryptosporidium parvum protozoa were not detected in the creek or the springs. Total culturable enteric viruses were detected in only one water sample taken from Rock Creek.

  2. 78 FR 36507 - Notice of Availability of a Treatment Evaluation Document; Methyl Bromide Fumigation of Blueberries

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-18

    ... schedule for methyl bromide fumigation of blueberries for Mediterranean fruit fly and South American fruit... explains why we have determined that it is effective at neutralizing these fruit flies. We are making the... risk from two fruit fly species, Ceratitis capitata (Mediterranean fruit fly, or Medfly) and...

  3. Methyl Bromide Alternatives Area-Wide Pest Management Project - South Atlantic Progress Report

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Protocols and Standard Operational Procedures (SOPs) were developed for collecting environmental and soil edaphic information during and after application of methyl bromide alternatives. Parameters measured included soil moisture, soil bulk density, percent moisture at field capacity (-0.3 bars wat...

  4. IDENTIFICATION OF NEW DRINKING WATER DISINFECTION BY-PRODUCTS FORMED IN THE PRESENCE OF BROMIDE

    EPA Science Inventory

    Using a combination of mass spectrometry and infrared spectroscopy, disinfection by-products (DBPs) were identified in ozonated drinking water containing elevated bromide levels, and in ozonated water treated with secondary chlorine or chloramine. Only one brominated by-product-d...

  5. Removal of methyl bromide in coastal seawater: Chemical and biological rates

    NASA Astrophysics Data System (ADS)

    King, Daniel B.; Saltzman, Eric S.

    1997-08-01

    A stable isotope tracer technique was used to investigate the loss rate of methyl bromide in surface ocean waters. Unfiltered and 0.2 μm-filtered or autoclaved aliquants of Biscayne Bay seawater samples were spiked with 13CH3Br at roughly 10-100 times ambient concentrations (50-800 pM) and incubated for 10-30 hours. The concentration of 13CH3Br was monitored using gas chromatography with isotope dilution mass spectrometry, with CD3Br as the isotope spike. Removal rates in unfiltered aliquants were significantly faster than in the 0.2 μm-filtered or autoclaved aliquants, indicating that some of the loss of methyl bromide was associated with particulate matter. Filtration experiments indicate that the particulate material responsible for methyl bromide loss is between 0.2 and 1.2 μm in diameter, suggesting that bacteria are likely to be responsible. The particulate-related removal of methyl bromide was inhibited by autoclaving, supporting a biological mechanism.

  6. Charging of silver bromide aqueous interface: evaluation of interfacial equilibrium constants from surface potential data.

    PubMed

    Preočanin, Tajana; Supljika, Filip; Kallay, Nikola

    2010-06-01

    A single crystal silver bromide electrode (SCr-AgBr) was used to measure the inner surface potential (Ψ(0)) at the silver bromide aqueous electrolyte interface as a function of the activities of Br(-) and Ag(+). Absolute values of the surface potential were calculated from electrode potentials of SCr-AgBr using the value of point of zero charge (pBr(pzc)=6.9 [H.A. Hoyen, R.M. Cole, J. Colloid Interface Sci. 41 (1972) 93.]) as the value of point of zero potential. Measurements were performed in potassium nitrate aqueous solutions. The Ψ(0)(pBr) function was linear and slightly dependent on the ionic strength. The reduction values of the slope with respect to the Nernst equation, expressed by the α coefficient, were 0.880,0.935, and 0.950 at ionic strengths of 10(-4), 10(-3), and 10(-2) mol dm(-3), respectively. The results were successfully interpreted by employing the surface complexation model, developed originally for metal oxides and adapted for silver halides. The thermodynamic ("intrinsic") equilibrium constants for binding of bromide (K(n)(∘)) and silver (K(p)(∘)) ions on the corresponding sites at the silver bromide surface were evaluated as lgK(n)(∘)=3.98; lgK(p)(∘)=2.48. Symmetrical counterion surface association was assumed and equilibrium constants were obtained as lgK(NO(3)(-))(∘)=lgK(K(+))(∘)=4.30.

  7. Chlorine utilization during trihalomethane formation in the presence of ammonia and bromide

    SciTech Connect

    Amy, G.L.; Chadik, P.A.; King, P.H.; Cooper, W.J.

    1984-10-01

    The chlorination of humic substances in the presence of ammonia and bromide and the resultant formation of trihalomethanes (THM) are described. The presence of ammonia substantially reduced but did not eliminate THM production during the chlorination of waters containing humic substances. The presence of bromide ion in waters containing humic substances leads to increased THM formation, both in the presence and in the absence of ammonia. In waters containing ammonia, bromide, and humic substances, brominated haloforms can form upon chlorination. Brominated species comprise a larger portion of the total THMs at higher chlorine doses within the free chlorine region of the breakpoint curve than at lower dose in the subbreakpoint region. Although the THM formation reaction is slow and may not be complete even after 1 week of reaction time, the initial rate of formation is competitive with the relatively rapid reactions of bromide oxidation and chloramine formation. After free chlorine is converted to chloramines, THM production should theoretically cease. THMs will form in parallel with chloramine formation. This behavior has important implications for water utilities that use a raw water source containing low but significant levels of ammonia such as the Biscayne Aquifer, or are considering chloramine disinfection as a THM control strategy.

  8. The use of agricultural by-products to capture methyl bromide following post-harvest fumigation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Activated carbons were prepared from plum and peach stone as well as almond and walnut shell and comparatively evaluated as sorbents to minimize the atmospheric emission of methyl bromide following postharvest fumigations. A variety of methods were used to make the activated carbons and each is desc...

  9. Experimental study of negative temperatures in lithium-bromide absorption refrigerating machines

    NASA Astrophysics Data System (ADS)

    Stepanov, K. I.; Mukhin, D. G.; Alekseenko, S. V.; Volkova, O. V.

    2015-07-01

    The authors have developed a method and presented experimental data for obtaining negative temperatures of evaporation in lithium-bromide absorption chillers driven by heat recovery. It has been found that the attainable temperature of the refrigerated medium is the value of -5 °C.

  10. Study of the antibacterial and antifungal activities of synthetic benzyl bromides, ketones, and corresponding chalcone derivatives

    PubMed Central

    Shakhatreh, Muhamad Ali K; Al-Smadi, Mousa L; Khabour, Omar F; Shuaibu, Fatima A; Hussein, Emad I; Alzoubi, Karem H

    2016-01-01

    Several applications of chalcones and their derivatives encouraged researchers to increase their synthesis as an alternative for the treatment of pathogenic bacterial and fungal infections. In the present study, chalcone derivatives were synthesized through cross aldol condensation reaction between 4-(N,N-dimethylamino)benzaldehyde and multiarm aromatic ketones. The multiarm aromatic ketones were synthesized through nucleophilic substitution reaction between 4-hydroxy acetophenone and benzyl bromides. The benzyl bromides, multiarm aromatic ketones, and corresponding chalcone derivatives were evaluated for their activities against eleven clinical pathogenic Gram-positive, Gram-negative bacteria, and three pathogenic fungi by the disk diffusion method. The minimum inhibitory concentration was determined by the microbroth dilution technique. The results of the present study demonstrated that benzyl bromide derivatives have strong antibacterial and antifungal properties as compared to synthetic chalcone derivatives and ketones. Benzyl bromides (1a and 1c) showed high ester activity against Gram-positive bacteria and fungi but moderate activity against Gram-negative bacteria. Therefore, these compounds may be considered as good antibacterial and antifungal drug discovery. However, substituted ketones (2a–b) as well as chalcone derivatives (3a–c) showed no activity against all the tested strains except for ketone (2c), which showed moderate activity against Candida albicans. PMID:27877017

  11. SYBR safe(TM) efficiently replaces ethidium bromide in Aspergillus fumigatus gene disruption.

    PubMed

    Canela, H M S; Takami, L A; Ferreira, M E S

    2017-02-08

    Invasive aspergillosis is a disease responsible for high mortality rates, caused mainly by Aspergillus fumigatus. The available drugs are limited and this disease continues to occur at an unacceptable frequency. Gene disruption is essential in the search for new drug targets. An efficient protocol for A. fumigatus gene disruption was described but it requires ethidium bromide, a genotoxic agent, for DNA staining. Therefore, the present study tested SYBR safe(TM), a non-genotoxic DNA stain, in A. fumigatus gene disruption protocol. The chosen gene was cipC, which has already been disrupted successfully in our laboratory. A deletion cassette was constructed in Saccharomyces cerevisiae and used in A. fumigatus transformation. There was no statistical difference between the tested DNA stains. The success rate of S. cerevisiae transformation was 63.3% for ethidium bromide and 70% for SYBR safe(TM). For A. fumigatus gene disruption, the success rate for ethidium bromide was 100 and 97% for SYBR safe(TM). In conclusion, SYBR safe(TM) efficiently replaced ethidium bromide, making this dye a safe and efficient alternative for DNA staining in A. fumigatus gene disruption.

  12. Steam disinfestation as a methyl bromide alternative in California cut flower nurseries

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Steam may be an effective alternative to methyl bromide in cut flower production in California. Advantages of steam include broad spectrum pest control and a zero hour re-entry interval. The principle disadvantage of sheet steaming is cost effectiveness due to current energy prices and application...

  13. Kinetics and mechanism of the oxidation of bromide by periodate in aqueous acidic solution.

    PubMed

    Szél, Viktor; Csekő, György; Horváth, Attila K

    2014-11-13

    The periodate–bromide reaction has been studied spectrophotometrically mainly in excess of bromide ion, monitoring the formation of the total amount of bromine at 450 nm at acidic buffered conditions and at a constant ionic strength in the presence of a phosphoric acid/dihydrogen phosphate buffer. The stoichiometry of the reaction was established to be strictly IO4(–) + 2Br(–) + 2H(+) → Br2 + IO3(–) + H2O. The formal kinetic order of the reactants was found to be perfectly one and two in the cases of periodate and bromide, respectively, but that of the hydrogen ion lies between one and two. We have also provided experimental evidence that dihydrogen phosphate accelerates the formation of bromine, suggesting the appearance of strong buffer assistance. On the basis of the experiments, a simple two-step kinetic model is proposed involving BrIO3 as a key intermediate that perfectly explains all of the experimental findings. Furthermore, we have also shown that in huge excess of bromide, the apparent rate coefficient obtained from the individual curve fitting method of the absorbance–time series is necessarily independent of the initial periodate concentration that may falsely be interpreted as the rate of bromine formation is also independent of the concentration of periodate.

  14. Ninety-Day Subchronic Oral Toxicity Study of Pyridostigmine Bromide in Rats. Volume 1

    DTIC Science & Technology

    1990-05-01

    myasthenia gravis because of its relative lack of untoward effects in comparison with other anticholinesterases (2). This relative lack of clinical...treatment of myasthenia gravis . Objecrive of Study The objective of this study was to determine the 90-day subchronic toxicity of pyridostigmine bromide in

  15. 77 FR 31564 - Notice of Availability of a Treatment Evaluation Document; Methyl Bromide Fumigation of Cottonseed

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-29

    ... Animal and Plant Health Inspection Service Notice of Availability of a Treatment Evaluation Document... add to the Plant Protection and Quarantine Treatment Manual a treatment schedule for methyl bromide... prepared a treatment evaluation document that describes the new treatment schedule and explains why we...

  16. 76 FR 34700 - Protection of Stratospheric Ozone: Request for Methyl Bromide Critical Use Exemption Applications...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-14

    ... AGENCY Protection of Stratospheric Ozone: Request for Methyl Bromide Critical Use Exemption Applications... FURTHER INFORMATION CONTACT: General Information: U.S. EPA Stratospheric Ozone Information Hotline, 1-800-296-1996; also http://www.epa.gov/ozone/mbr . Technical Information: Bill Chism, U.S....

  17. Methyl bromide release from activated carbon and the soil/water/carbon interface

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methyl Bromide (MB) is a major source of stratospheric bromine radical, a known depletor of ozone. The use of ozone-depleting chemicals, including MB, is regulated by the Montreal Protocol. Critical uses of MB are permitted, such as when postharvest fumigation is mandated by an importing country. Fo...

  18. EVALUATION OF CONTAINMENT AND CONTROL OPTIONS FOR METHYL BROMIDE IN COMMODITY TREATMENT

    EPA Science Inventory

    The report gives results of an investigation of means for methyl bromide (MeBr) recovery, reuse, and destruction to prevent atmospheric emissions if its limited use were still allowed. (NOTE: MeBr is an ozone-depleting chemical scheduled to be phased out by the Clean Air Act by t...

  19. Reduced rates and alternatives to methyl bromide for snapdragon production in Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A field trial was conducted to evaluate soil solarization, Midas™ (iodomethane:chloropicrin 50:50, Arysta LifeScience Corp., Cary, NC) and different rates and formulations of methyl bromide under standard and metalized films for the production of snapdragon (Antirrhinum majus) in Martin County, Flor...

  20. Highly efficient synthesis of phenols by copper-catalyzed hydroxylation of aryl iodides, bromides, and chlorides.

    PubMed

    Yang, Kai; Li, Zheng; Wang, Zhaoyang; Yao, Zhiyi; Jiang, Sheng

    2011-08-19

    8-Hydroxyquinolin-N-oxide was found to be a very efficient ligand for the copper-catalyzed hydroxylation of aryl iodides, aryl bromides, or aryl chlorides under mild reaction conditions. This methodology provides a direct transformation of aryl halides to phenols and to alkyl aryl ethers. The inexpensive catalytic system showed great functional group tolerance and excellent selectivity.

  1. Practical Cross-Coupling between O-Based Electrophiles and Aryl Bromides via Ni Catalysis.

    PubMed

    Cao, Zhi-Chao; Luo, Qin-Yu; Shi, Zhang-Jie

    2016-12-02

    Cross-coupling of various O-based electrophiles with aryl bromides was developed through Ni-catalyzed C-O activation in the presence of magnesium. Beside carboxylates, carbamates, and ethers, phenols exhibited excellent reactivity under modified conditions. This chemistry was featured as a simple and environmentally benign process with low catalyst loading and easy manipulations. The method exhibited broad substrate scopes.

  2. Efficient palladium-catalyzed coupling reactions of aryl bromides and chlorides with phenols.

    PubMed

    Hu, Tongjie; Schulz, Thomas; Torborg, Christian; Chen, Xiaorong; Wang, Jun; Beller, Matthias; Huang, Jun

    2009-12-21

    A convenient and general palladium-catalyzed coupling reaction of aryl bromides and chlorides with phenols was developed. Various functional groups such as nitriles, aldehydes, ketones and esters are well tolerated and the corresponding products are obtained in good to excellent yield.

  3. Synthesis of Vinyl Trifluoromethyl Thioethers via Copper-Mediated Trifluoromethylthiolation of Vinyl Bromides.

    PubMed

    Huang, Yangjie; Ding, Jianping; Wu, Chuyi; Zheng, Huidong; Weng, Zhiqiang

    2015-03-06

    A copper-mediated trifluoromethylthiolation of vinyl bromides has been developed. This method provides ready access to vinyl trifluoromethyl thioethers in good to high yields from simple, inexpensive starting materials. A broad substrate scope is achieved, and the reaction is compatible with various functional groups, including cyano, nitro, trifluoromethyl, alkoxy, amino, halide, and heterocyclic groups.

  4. Effect of increasing bromide concentration on toxicity in treated drinking water.

    PubMed

    Sawade, Emma; Fabris, Rolando; Humpage, Andrew; Drikas, Mary

    2016-04-01

    Research is increasingly indicating the potential chronic health effects of brominated disinfection by-products (DBPs). This is likely to increase with elevated bromide concentrations resulting from the impacts of climate change, projected to include extended periods of drought and the sudden onset of water quality changes. This will demand more rigorous monitoring throughout distribution systems and improved water quality management at water treatment plants (WTPs). In this work the impact of increased bromide concentration on formation of DBPs following conventional treatment and chlorination was assessed for two water sources. Bioanalytical tests were utilised to determine cytotoxicity of the water post disinfection. Coagulation was shown to significantly reduce the cytotoxicity of the water, indicating that removal of natural organic matter DBP precursors continues to be an important factor in drinking water treatment. Most toxic species appear to form within the first half hour following disinfectant addition. Increasing bromide concentration across the two waters was shown to increase the formation of trihalomethanes and shifted the haloacetic acid species distribution from chlorinated to those with greater bromine substitution. This correlated with increasing cytotoxicity. This work demonstrates the challenges faced by WTPs and the possible effects increasing levels of bromide in source waters could have on public health.

  5. Methods to facilitate the adoption of alternatives to methyl bromide soil fumigation by California strawberry growers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The goal of this project is to facilitate the adoption of strawberry production systems that do not use methyl bromide (MB). The five year project initially focused on fumigant alternatives to MB. The project has resulted in increased use of barrier films that reduce fumigant emission. The focus s...

  6. TRIBROMOPYRROLE, BROMINATED ACIDS, AND OTHER DISINFECTION BYPRODUCTS PRODUCED BY DISINFECTION OF DRINKING WATER RICH IN BROMIDE

    EPA Science Inventory

    Using gas chromatography/mass spectrometry (GC/MS), we investigated the formation of disinfection byproducts (DBPs) from high bromide waters (2 mg/L) treated with chlorine or chlorine dioxide used in combination with chlorine and chloramines. This study represents the first comp...

  7. Neuropsychological Functioning in Gulf War Veterans Exposed to Pesticides and Pyridostigmine Bromide

    DTIC Science & Technology

    2008-08-01

    this sweeping pesticide review, the EPA also suggested that some OP pesticides may have endocrine disrupting properties at doses much lower than would...EPA because of its persistence in the environment, ability to bioaccumulate, potential as a carcinogen and evidence as an endocrine disruptor (http...War Veterans Exposed to Pesticides and Pyridostigmine Bromide PRINCIPAL INVESTIGATOR: Maxine Krengel, Ph.D. CONTRACTING

  8. 78 FR 32646 - Protection of Stratospheric Ozone: Request for Methyl Bromide Critical Use Exemption Applications...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-31

    ... on Substances that Deplete the Ozone Layer for industrialized countries and to allow EPA to provide a... AGENCY Protection of Stratospheric Ozone: Request for Methyl Bromide Critical Use Exemption Applications... FURTHER INFORMATION CONTACT: General Information: U.S. EPA Stratospheric Ozone Information Hotline,...

  9. 77 FR 29341 - Protection of Stratospheric Ozone: Request for Methyl Bromide Critical Use Exemption Applications...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-17

    ... to the provisions of the Montreal Protocol on Substances that Deplete the Ozone Layer for... AGENCY Protection of Stratospheric Ozone: Request for Methyl Bromide Critical Use Exemption Applications... FURTHER INFORMATION CONTACT: General Information: U.S. EPA Stratospheric Ozone Information Hotline,...

  10. Headspace gas chromatographic method for determination of methyl bromide in food ingredients

    SciTech Connect

    DeVries, J.W.; Broge, J.M.; Schroeder, J.P.; Bowers, R.H.; Larson, P.A.; Burns, N.M.

    1985-11-01

    A headspace gas chromatographic (GC) method, which can be automated, has been developed for determination of methyl bromide. This method has been applied to wheat, flour, cocoa, and peanuts. Samples to be analyzed are placed in headspace sample vials, water is added, and the vials are sealed with Teflon-lined septa. After an appropriate equilibration time at 32 degrees C, the samples are analyzed within 10 h. A sample of the headspace is withdrawn and analyzed on a gas chromatograph equipped with an electron capture detector (ECD). Methyl bromide levels were quantitated by comparison of peak area with a standard. The standard was generated by adding a known amount of methyl bromide to a portion of the matrix being analyzed and which was known to be methyl bromide free. The detection limit of the method was 0.4 ppb. The coefficient of variation (CV) was 6.5% for wheat, 8.3% for flour, 3.3% for cocoa, and 11.6% for peanuts.

  11. OZONATION BYPRODUCTS: IDENTIFICATION OF BROMOHYDRINS FROM THE OZONATION OF NATURAL WATERS WITH ENHANCED BROMIDE LEVELS

    EPA Science Inventory

    When ozone is used in the treatment of drinking water, it reacts with both inorganic and organic compounds to form byproducts. f bromide is present, it may be oxidized to hydrobromous acid, which may than react with natural organic matter to form brominated organic compounds. he ...

  12. A novel explicit approach to model bromide and pesticide transport in connected soil structures

    NASA Astrophysics Data System (ADS)

    Klaus, J.; Zehe, E.

    2011-07-01

    The present study tests whether an explicit treatment of worm burrows and tile drains as connected structures is feasible for simulating water flow, bromide and pesticide transport in structured heterogeneous soils at hillslope scale. The essence is to represent worm burrows as morphologically connected paths of low flow resistance in a hillslope model. A recent Monte Carlo study (Klaus and Zehe, 2010, Hydrological Processes, 24, p. 1595-1609) revealed that this approach allowed successful reproduction of tile drain event discharge recorded during an irrigation experiment at a tile drained field site. However, several "hillslope architectures" that were all consistent with the available extensive data base allowed a good reproduction of tile drain flow response. Our second objective was thus to find out whether this "equifinality" in spatial model setups may be reduced when including bromide tracer data in the model falsification process. We thus simulated transport of bromide for the 13 spatial model setups that performed best with respect to reproduce tile drain event discharge, without any further calibration. All model setups allowed a very good prediction of the temporal dynamics of cumulated bromide leaching into the tile drain, while only four of them matched the accumulated water balance and accumulated bromide loss into the tile drain. The number of behavioural model architectures could thus be reduced to four. One of those setups was used for simulating transport of Isoproturon, using different parameter combinations to characterise adsorption according to the Footprint data base. Simulations could, however, only reproduce the observed leaching behaviour, when we allowed for retardation coefficients that were very close to one.

  13. Kinetics of Chlorination of Benzophenone-3 in the Presence of Bromide and Ammonia.

    PubMed

    Abdallah, Pamela; Deborde, Marie; Dossier Berne, Florence; Karpel Vel Leitner, Nathalie

    2015-12-15

    The aim of this study was to assess the impact of chlorination on the degradation of one of the most commonly used UV filters (benzophenone-3 (BP-3)) and the effects of bromide and ammonia on the kinetics of BP-3 elimination. Bromide and ammonia are rapidly converted to bromine and chloramines during chlorination. At first, the rate constants of chlorine, bromine and monochloramine with BP-3 were determined at various pH levels. BP-3 was found to react rapidly with chlorine and bromine, with values of apparent second order rate constants equal to 1.25(±0.14) × 10(3) M(-1)·s(-1) and 4.04(±0.54) × 10(6) M(-1)·s(-1) at pH 8.5 for kChlorine/BP-3 and kBromine/BP-3, respectively, whereas low monochloramine reactivity was observed (kNH2Cl/BP-3 = 0.112 M(-1)·s(-1)). To assess the impact of the inorganic content of water on BP-3 degradation, chlorination experiments with different added concentrations of bromide and/or ammonia were conducted. Under these conditions, BP-3 degradation was found to be enhanced in the presence of bromide due to the formation of bromine, whereas it was inhibited in the presence of ammonia. However, the results obtained were pH dependent. Finally, a kinetic model considering 18 reactions was developed using Copasi to estimate BP-3 degradation during chlorination in the presence of bromide and ammonia.

  14. Utility of Bromide and Heat Tracers for Aquifer Characterization Affected by Highly Transient Flow Conditions

    SciTech Connect

    Ma, Rui; Zheng, Chunmiao; Zachara, John M.; Tonkin, Matthew J.

    2012-08-29

    A tracer test using both bromide and heat tracers conducted at the Integrated Field Research Challenge site in Hanford 300 Area (300A), Washington, provided an instrument for evaluating the utility of bromide and heat tracers for aquifer characterization. The bromide tracer data were critical to improving the calibration of the flow model complicated by the highly dynamic nature of the flow field. However, most bromide concentrations were obtained from fully screened observation wells, lacking depth-specific resolution for vertical characterization. On the other hand, depth-specific temperature data were relatively simple and inexpensive to acquire. However, temperature-driven fluid density effects influenced heat plume movement. Moreover, the temperature data contained “noise” caused by heating during fluid injection and sampling events. Using the hydraulic conductivity distribution obtained from the calibration of the bromide transport model, the temperature depth profiles and arrival times of temperature peaks simulated by the heat transport model were in reasonable agreement with observations. This suggested that heat can be used as a cost-effective proxy for solute tracers for calibration of the hydraulic conductivity distribution, especially in the vertical direction. However, a heat tracer test must be carefully designed and executed to minimize fluid density effects and sources of noise in temperature data. A sensitivity analysis also revealed that heat transport was most sensitive to hydraulic conductivity and porosity, less sensitive to thermal distribution factor, and least sensitive to thermal dispersion and heat conduction. This indicated that the hydraulic conductivity remains the primary calibration parameter for heat transport.

  15. Comparison of salbutamol and ipratropium bromide versus salbutamol alone in the treatment of acute severe asthma.

    PubMed

    Hossain, A S; Barua, U K; Roy, G C; Sutradhar, S R; Rahman, I; Rahman, G

    2013-04-01

    The use of nebulized Ipratropium bromide, quaternary anticholinergic bronchodilators in combination with beta-agonist for the treatment of acute asthma in adults is controversial. In a view of different recommendation the present study is undertaken in Bangladeshi patients. Combination of inhaled Ipratropium bromide and Salbutamol provides greater bronchodilatation than mono therapy with Salbutamol alone in acute severe asthma. Patients of severe asthma (PEFR <50% of predicted) were enrolled into control group (Salbutamol only) and case (Salbutamol + Ipratropium bromide) group. After measurement of peak expiratory flow, patient received 3 doses of 2.5 mg Salbutamol (n=40) only or 3 doses of both 2.5mg Salbutamol and 500mcg Ipratropium bromide at an interval of 20 minutes (n=40) through a jet nebulizer. Peak flow was reassessed 30 & 60 minutes after treatment. Peak flow at baseline was similar in two groups. Then at 30 minutes after nebulization, the mean±SD percentage increase in peak flow was greater in combination group (60.01±35.01%) than Salbutamol group (44.47±25.03%) with difference of 16% (p=0.025). At 60 minutes the percentage increase in peak flow was about 32% greater in combination group than Salbutamol group (94.44±33.70% vs. 62.57±29.26%, p=0.000) and combination group reached percentage predicted peak flow more than 60% while Sabutamol group did not. Ipratropium Bromide and Salbutamol nebulized combinedly have better bronchodilating effect than Salbultamol alone in acute severe asthma.

  16. Improvement of Quality of Nonanesthetic Colonoscopy by Preoperative Administration of Pinaverium Bromide

    PubMed Central

    Wang, Xiao-Lin; Zhou, Jian-Ning; Ren, Li; Pan, Xiao-Li; Ren, Hong-Yu; Liu, Jun

    2017-01-01

    Background: Nonanesthetic colonoscopy is popular in clinical practice in China. However, intestinal spasms often result in a prolonged examination time, increased operating difficulties, decreased polyp detection rate, and failure to complete the procedure clinically. Therefore, exploring alternative approaches that can reduce the pain in patients during colonoscopy is of utmost importance, and finding the optimal preoperative administration to improve the quality of nonanesthetic colonoscopy is also necessary. This study aimed to investigate the effects of the prophylactic administration of pinaverium bromide before colonoscopy and the effects of pinaverium bromide alone at different time points or combined with scopolamine butylbromide. Methods: A randomized controlled trial was performed on a cohort of 1000 patients who underwent colonoscopy in outpatient clinic of Wuhan Union Hospital. The patients were randomly assigned to the following groups: Group A, given oral pinaverium bromide (100 mg, three times a day) one day before examination combined with intramuscular injection of scopolamine butylbromide (20 mg) 10 min before colonoscopy; Group B0, given pinaverium bromide alone on the day of colonoscopy (100 mg, three times a day); Group B1, given pinaverium bromide alone (100 mg, three times a day) one day before colonoscopy; Group B2, given pinaverium bromide alone (100 mg, three times a day) two days before colonoscopy; and Group C, given scopolamine butylbromide alone (20 mg) before colonoscopy. The successful rate of colonoscopy, procedure time, degree of abdominal pain, and polyp detection rate were recorded and compared among all groups. Results: The successful rate of colonoscopy in Group B1 (82.0%) and Group B2 (83.0%) was significantly higher than that in Group B0 (62.0%, all P < 0.01). The time to reach the ileocecal region in Group B1 and Group B2 were lower than those in Group B0 (all P < 0.05). However, no significant differences were observed in

  17. Severe Illness from Methyl Bromide Exposure at a Condominium Resort--U.S. Virgin Islands, March 2015.

    PubMed

    Kulkarni, Prathit A; Duncan, Mary Anne; Watters, Michelle T; Graziano, Leah T; Vaouli, Elena; Cseh, Larry F; Risher, John F; Orr, Maureen F; Hunte-Ceasar, Tai C; Ellis, Esther M

    2015-07-24

    On March 22, 2015, the Agency for Toxic Substances and Disease Registry (ATSDR) was notified by the U.S. Environmental Protection Agency (EPA) of four cases of suspected acute methyl bromide toxicity among family members vacationing at a condominium resort in the U.S. Virgin Islands. Methyl bromide is a pesticide that has been banned in the United States for use in homes and other residential settings. An investigation conducted by the U.S. Virgin Islands Department of Health (VIDOH), the U.S. Virgin Islands Department of Planning and Natural Resources (DPNR), and EPA confirmed that methyl bromide had been used as a fumigant on March 18 in the building where the family had been residing, 2 days before they were transported to the hospital; three family members had life-threatening illness. On March 25, 2015, a stop-use order for methyl bromide was issued by DPNR to the pest control company that had performed the fumigation. Subsequent investigation revealed that previous fumigation with methyl bromide had occurred on October 20, 2014, at the same condominium resort. In addition to the four ill family members, 37 persons who might have been exposed to methyl bromide as a result of the October 2014 or March 2015 fumigations were identified by VIDOH and ATSDR. Standardized health questionnaires were administered to 16 of the 20 persons for whom contact information was available; six of 16 had symptoms consistent with methyl bromide exposure, including headache and fatigue. Pest control companies should be aware that use of methyl bromide is banned in homes and other residential settings, and clinicians should be aware of the toxicologic syndrome that exposure to methyl bromide can cause.

  18. Bromate formation in bromide-containing water through the cobalt-mediated activation of peroxymonosulfate.

    PubMed

    Li, Zhaobing; Chen, Zhi; Xiang, Yingying; Ling, Li; Fang, Jingyun; Shang, Chii; Dionysiou, Dionysios D

    2015-10-15

    Bromate formation in bromide-containing water through the cobalt (Co)-mediated activation of peroxymonosulfate (PMS) was investigated. Increasing the PMS dosage and the cobalt dosage increased the formation of bromate and bromate yields of up to 100% were recorded under the test conditions. The bromate yield increased to a maximum as the pH rose from 2.7 to 6 before decreasing by over 90% as the pH rose further from 6 to above 9. The bromate formation is a two-step process involving free bromine as a key intermediate and bromate as the final product. In the first step, apart from the known oxidation of bromide to free bromine and of free bromine to bromate by sulfate radicals (SO4(-)), Co(III) produced from the oxidation of Co(II) by PMS and SO4(-) also oxidizes bromide to free bromine. The contribution of Co(III) to the bromate formation was verified with the addition of methanol and EDTA, a radical scavenger and a Co(III) ligand, respectively. In the presence of methanol, free bromine formation increased with increasing Co(II) dosage but no bromate was detected, indicating that Co(III) oxidized bromide to form free bromine but not bromate. In the presence of both EDTA and methanol, no free bromine or bromate was detected, as Co(III) was stabilized by EDTA to form the Co(III)EDTA(-) complex, which could not oxidize bromide. Mathematical simulation further suggested that Co(III) outweighed SO4(-) to oxidize bromide to free bromine. On the other hand, SO4(-) is essential for the oxidation of free bromine to bromate in the second step. In real water, the presence of NOM significantly decreased the bromate formation but caused the brominated organic DBP formation with high quantity. This is the first study to demonstrate the significant bromate formation in the Co/PMS system and the substantial contribution of Co(III) to the formation.

  19. Atmospheric volatilization of methyl bromide, 1,3-dichloropropene, and propargyl bromide through two plastic films: transfer coefficient and temperature effect

    NASA Astrophysics Data System (ADS)

    Wang, D.; Yates, S. R.; Gan, J.; Knuteson, J. A.

    Atmospheric emission of methyl bromide (MeBr) and its potential alternative chemicals such as 1,3-dichloropropene (1,3-D) and propargyl bromide (PrBr) can contribute to air pollution and ozone depletion (for MeBr). One of the main sources of these chemicals is from agricultural soil fumigation. To understand the volatilization dynamics, emission of MeBr, 1,3-D, and PrBr through a polyethylene-based high-barrier film (HBF) and a virtually impermeable film (VIF) was measured using an air flow and sampling system that produced >90% mass balance. The experiment was conducted outdoors and was subjected to ambient daily temperature variations. The HBF film was found to be very permeable to 1,3-D and PrBr, but somewhat less permeable to MeBr. The VIF film was very impermeable to 1,3-D, PrBr, or MeBr. Measured volatilization flux, in general, exhibited strong diurnal variations which were controlled by film temperature. Unlike the HBF film, a time lag (˜12 h) was observed between high-temperatures and high-emission flux values for the VIF film. An impermeable film may be used as an effective means of controlling the atmospheric emission of MeBr and its alternative chemicals.

  20. Underpotential deposition of Cu on Pt(001): Interface structure and the influence of adsorbed bromide

    NASA Astrophysics Data System (ADS)

    Lucas, C. A.; Markovic, N. M.; Ross, P. N.

    1998-05-01

    Using in situ x-ray diffraction, we studied the underpotential deposition (UPD) of copper onto a Pt(001) electrode both in pure perchloric acid and in the presence of bromide anions. In pure perchloric acid, the Cu is deposited in pseudomorphic p(1×1) islands. In the presence of bromide anions, the strong Pt-Br interaction significantly broadens the potential range of Cu UPD. We propose that Br remains in the interface region throughout the UPD process, at first in a disordered Cu-Br phase and then, at more negative potential, forming a c(2×2) closed-packed monolayer on top of the completed p(1×1) Cu monolayer. The structures are compared to those found during Cu UPD onto Pt(111), and explained in terms of the metal-halide interactions and the Pt surface atomic geometry.