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Sample records for vitro screening cascade

  1. A screening cascade to identify ERβ ligands.

    Science.gov (United States)

    Filgueira, Carly S; Benod, Cindy; Lou, Xiaohua; Gunamalai, Prem S; Villagomez, Rosa A; Strom, Anders; Gustafsson, Jan-Åke; Berkenstam, Anders L; Webb, Paul

    2014-01-01

    The establishment of effective high throughput screening cascades to identify nuclear receptor (NR) ligands that will trigger defined, therapeutically useful sets of NR activities is of considerable importance. Repositioning of existing approved drugs with known side effect profiles can provide advantages because de novo drug design suffers from high developmental failure rates and undesirable side effects which have dramatically increased costs. Ligands that target estrogen receptor β (ERβ) could be useful in a variety of diseases ranging from cancer to neurological to cardiovascular disorders. In this context, it is important to minimize cross-reactivity with ERα, which has been shown to trigger increased rates of several types of cancer. Because of high sequence similarities between the ligand binding domains of ERα and ERβ, preferentially targeting one subtype can prove challenging. Here, we describe a sequential ligand screening approach comprised of complementary in-house assays to identify small molecules that are selective for ERβ. Methods include differential scanning fluorimetry, fluorescence polarization and a GAL4 transactivation assay. We used this strategy to screen several commercially-available chemical libraries, identifying thirty ERβ binders that were examined for their selectivity for ERβ versus ERα, and tested the effects of selected ligands in a prostate cancer cell proliferation assay. We suggest that this approach could be used to rapidly identify candidates for drug repurposing.

  2. Cascade Screening in Families with Inherited Cardiac Diseases Driven by Cardiologists

    DEFF Research Database (Denmark)

    Theilade, Juliane; Kanters, Jørgen; Henriksen, Finn Lund

    2013-01-01

    We assessed the outcome of cascade screening of families with congenital long QT syndrome (LQTS) in Danish heart centers.......We assessed the outcome of cascade screening of families with congenital long QT syndrome (LQTS) in Danish heart centers....

  3. Preliminary phytochemical screening and in vitro antimicrobial ...

    African Journals Online (AJOL)

    The petroleum ether extract of Croton zambesicus Muell Arg. was subjected to preliminary Phytochemical screening and in vitro antimicrobial tests. The Phytochemical tests were conducted using standard methods of analysis and the extract revealed the presence of cardiac glycosides and steroids. Antimicrobial effects of ...

  4. In vitro fertilization with preimplantation genetic screening

    NARCIS (Netherlands)

    Mastenbroek, Sebastiaan; Twisk, Moniek; van Echten-Arends, Jannie; Sikkema-Raddatz, Birgit; Korevaar, Johanna C.; Verhoeve, Harold R.; Vogel, Niels E. A.; Arts, Eus G. J. M.; de Vries, Jan W. A.; Bossuyt, Patrick M.; Buys, Charles H. C. M.; Heineman, Maas Jan; Repping, Sjoerd; van der Veen, Fulco

    2007-01-01

    BACKGROUND: Pregnancy rates in women of advanced maternal age undergoing in vitro fertilization (IVF) are disappointingly low. It has been suggested that the use of preimplantation genetic screening of cleavage-stage embryos for aneuploidies may improve the effectiveness of IVF in these women.

  5. In vitro toxicity screening of colored smokes

    NARCIS (Netherlands)

    Langenberg, J.P.; Alblas, M.J.; Makkus, J.C.; Meer, J.A. van der; Mars-Groenendijk, R.H.; Groeneveld, F.R.; Norbert, M.; Lingen, J.N.J. van

    2009-01-01

    Data on the acute and/or long-term toxicity of colored smokes appear to be scarce and inconsistent. Therefore, the objective of this study is to obtain more insight on this matter. For this purpose, existing platforms for in vitro toxicity screening are evaluated with respect to their applicability

  6. Using the Cascade Model to Improve Antenatal Screening for the Hemoglobin Disorders

    Science.gov (United States)

    Gould, Dinah; Papadopoulos, Irena; Kelly, Daniel

    2012-01-01

    Introduction: The inherited hemoglobin disorders constitute a major public health problem. Facilitators (experienced hemoglobin counselors) were trained to deliver knowledge and skills to "frontline" practitioners to enable them to support parents during antenatal screening via a cascade (train-the-trainer) model. Objectives of…

  7. In vitro screening of selected essential oils from medicinal plants ...

    African Journals Online (AJOL)

    , Cymbopogon citratus, Eucalyptus citriodora, Laurus nobilis, Lippia multiflora, Mentha piperita, Ocimum basilicum, Ocimum gratissimum and Zingiber officinalis on enteric methane production in in vitro batch cultures screening experiments ...

  8. Phytochemical Screening and In vitro Evaluation of Pharmacological ...

    African Journals Online (AJOL)

    Phytochemical Screening and In vitro Evaluation of Pharmacological Activities of Aphanamixis polystachya (Wall) Parker Fruit Extracts. AS Apu, FA Chowdhury, F Khatun, ATM Jamaluddin, AH Pathan, A Pal ...

  9. In vitro antioxidant activity and phytochemical screening of methanol ...

    African Journals Online (AJOL)

    In this study, phytochemical screening and in vitro antioxidant activity of methanol extracts of D. edulis and F. capensis leaves were evaluated. Each plant leaves were extracted in methanol using standard procedures. The phytochemical screening of the resulting extracts showed the presence of cardiac glycosides, ...

  10. Long-Term Cardiovascular Risk in Heterozygous Familial Hypercholesterolemia Relatives Identified by Cascade Screening

    DEFF Research Database (Denmark)

    Kjærgaard, Kasper Aalbæk; Christiansen, Morten Krogh; Schmidt, Morten

    2017-01-01

    with a low-density lipoprotein receptor (LDLR) mutation who were all recommended statin therapy. METHODS AND RESULTS: Participants were identified by cascade screening at Aarhus University Hospital during 1992-1994. A comparison cohort from the Danish general population was matched 10:1 to relatives by birth...... statins during their follow-up period. Despite frequent use of lipid-lowering medication, the adjusted hazard ratio of the primary end point was 1.65 (95% confidence interval, 1.17-2.33) in mutation-carrying relatives compared with the general population cohort. The risk in non-mutation-carrying relatives...... was not different from that of the general population cohort (adjusted hazard ratio: 0.85; 95% confidence interval, 0.56-1.29). Comparing mutation-carrying relatives with non-mutation-carrying relatives, the adjusted hazard ratio was 1.94 (95% confidence interval, 1.14-3.31). Results were driven by nonfatal events...

  11. Low-density lipoprotein receptor gene mutations and cardiovascular risk in a large genetic cascade screening population

    NARCIS (Netherlands)

    Umans-Eckenhausen, Marina A. W.; Sijbrands, Eric J. G.; Kastelein, John J. P.; Defesche, Joep C.

    2002-01-01

    Background-A large cohort of patients with familial hypercholesterolemia (FH), free from selection for cardiovascular disease (CVD), and their unaffected relatives was collected by genetic cascade screening and examined for the influence of different mutations of the LDL receptor gene on lipoprotein

  12. Phytochemical screening and in vitro acetylcholinesterase inhibitory ...

    African Journals Online (AJOL)

    Recent studies have shown the effectiveness of plants as enhancers of memory activity. This study sought to investigate the inhibitory effect of seven plants on acetylcholinesterase and its phytochemical contents. The in vitro acetylcholinesterase inhibitory effect by the seven plants and their phytochemical contents each, ...

  13. In Vitro screening of tomato genotypes for drought resistance using ...

    African Journals Online (AJOL)

    Drought is a major abiotic factor that limits plant growth and productivity. Tomato is an important vegetable crop and area under production is limited by irrigation water scarcity. Effort was made to screen tomato germplasm under in vitro condition using polyethylene glycol (PEG) at four concentrations (0, 20, 40 and 60 g/l) ...

  14. Phytochemical Screening and In Vitro Antitrypanosomal Activity of ...

    African Journals Online (AJOL)

    African trypanosomiasis is a major public health problem having serious economic implications affecting both human and agricultural development. The present work dealt with phytochemical screening and in vitro antitrypanosomal activity testing of the hydromethanolic and dichloromethane (DCM) extracts of the aerial ...

  15. Steroidogenesis in vitro : towards relevant models for endocrine disruptor screening

    NARCIS (Netherlands)

    Roelofs, M.J.E.|info:eu-repo/dai/nl/357301137

    2016-01-01

    Starting our search for in vitro alternative methods to screen for steroidogenesis toxicity, we focused on the effects of (suggested) endocrine disrupting compounds (EDCs) on cytochrome P450 17 (CYP17) enzyme activity. CYP17 is responsible for conversion of progestagens to dehydroepiandrosterone

  16. Phytochemical Screening and In vitro Evaluation of Pharmacological ...

    African Journals Online (AJOL)

    HP

    Purpose: To investigate the crude n-hexane, ethyl acetate and methanol extracts of Aphanamixis polystachya fruit for their cytotoxic, antimicrobial, antioxidant and thrombolytic activities. Methods: The fruit extracts were screened for major phytochemical compounds using in vitro established procedures. Antimicrobial and ...

  17. In vitro screening of selected herbicides on rhizosphere mycoflora ...

    African Journals Online (AJOL)

    In vitro screening of selected herbicides on rhizosphere mycoflora from yellow pepper ( Capsicum annum L var. Nsukka yellow) ... Fr) Lind, Trichoderma harmatum Bain aggr; Aspergillus niger Van Tiegh; Sclerotium rolfsii Sacc; Penicillium sp; Alternaria sp; Fusarium solani (Mart) Sacc and Rhizoctonia sp. The results ...

  18. In Vitro High Throughput Screening, What Next? Lessons from the Screening for Aurora Kinase Inhibitors

    Directory of Open Access Journals (Sweden)

    Thi-My-Nhung Hoang

    2014-02-01

    Full Text Available Based on in vitro assays, we performed a High Throughput Screening (HTS to identify kinase inhibitors among 10,000 small chemical compounds. In this didactic paper, we describe step-by-step the approach to validate the hits as well as the major pitfalls encountered in the development of active molecules. We propose a decision tree that could be adapted to most in vitro HTS.

  19. In vitro high throughput screening, what next? Lessons from the screening for aurora kinase inhibitors.

    Science.gov (United States)

    Hoang, Thi-My-Nhung; Vu, Hong-Lien; Le, Ly-Thuy-Tram; Nguyen, Chi-Hung; Molla, Annie

    2014-02-27

    Based on in vitro assays, we performed a High Throughput Screening (HTS) to identify kinase inhibitors among 10,000 small chemical compounds. In this didactic paper, we describe step-by-step the approach to validate the hits as well as the major pitfalls encountered in the development of active molecules. We propose a decision tree that could be adapted to most in vitro HTS.

  20. Modelling of the Blood Coagulation Cascade in an In Vitro Flow System

    DEFF Research Database (Denmark)

    Andersen, Nina Marianne; Sørensen, Mads Peter; Efendiev, Messoud A.

    2010-01-01

    We derive a mathematical model of a part of the blood coagulation cascade set up in a perfusion experiment. Our purpose is to simulate the influence of blood flow and diffusion on the blood coagulation pathway. The resulting model consists of a system of partial differential equations taking...... and flow equations, which guarantee non negative concentrations at all times. The criteria is applied to the model of the blood coagulation cascade....

  1. Identification of Trypanocidal Activity for Known Clinical Compounds Using a New Trypanosoma cruzi Hit-Discovery Screening Cascade.

    Science.gov (United States)

    De Rycker, Manu; Thomas, John; Riley, Jennifer; Brough, Stephen J; Miles, Tim J; Gray, David W

    2016-04-01

    Chagas disease is a significant health problem in Latin America and the available treatments have significant issues in terms of toxicity and efficacy. There is thus an urgent need to develop new treatments either via a repurposing strategy or through the development of new chemical entities. A key first step is the identification of compounds with anti-Trypanosoma cruzi activity from compound libraries. Here we describe a hit discovery screening cascade designed to specifically identify hits that have the appropriate anti-parasitic properties to warrant further development. The cascade consists of a primary imaging-based assay followed by newly developed and appropriately scaled secondary assays to predict the cidality and rate-of-kill of the compounds. Finally, we incorporated a cytochrome P450 CYP51 biochemical assay to remove compounds that owe their phenotypic response to inhibition of this enzyme. We report the use of the cascade in profiling two small libraries containing clinically tested compounds and identify Clemastine, Azelastine, Ifenprodil, Ziprasidone and Clofibrate as molecules having appropriate profiles. Analysis of clinical derived pharmacokinetic and toxicity data indicates that none of these are appropriate for repurposing but they may represent suitable start points for further optimisation for the treatment of Chagas disease.

  2. Identification of Trypanocidal Activity for Known Clinical Compounds Using a New Trypanosoma cruzi Hit-Discovery Screening Cascade.

    Directory of Open Access Journals (Sweden)

    Manu De Rycker

    2016-04-01

    Full Text Available Chagas disease is a significant health problem in Latin America and the available treatments have significant issues in terms of toxicity and efficacy. There is thus an urgent need to develop new treatments either via a repurposing strategy or through the development of new chemical entities. A key first step is the identification of compounds with anti-Trypanosoma cruzi activity from compound libraries. Here we describe a hit discovery screening cascade designed to specifically identify hits that have the appropriate anti-parasitic properties to warrant further development. The cascade consists of a primary imaging-based assay followed by newly developed and appropriately scaled secondary assays to predict the cidality and rate-of-kill of the compounds. Finally, we incorporated a cytochrome P450 CYP51 biochemical assay to remove compounds that owe their phenotypic response to inhibition of this enzyme. We report the use of the cascade in profiling two small libraries containing clinically tested compounds and identify Clemastine, Azelastine, Ifenprodil, Ziprasidone and Clofibrate as molecules having appropriate profiles. Analysis of clinical derived pharmacokinetic and toxicity data indicates that none of these are appropriate for repurposing but they may represent suitable start points for further optimisation for the treatment of Chagas disease.

  3. In Vitro Screening for Cytotoxic Activity of Herbal Extracts

    Directory of Open Access Journals (Sweden)

    Valter R. M. Lombardi

    2017-01-01

    Full Text Available Experimental studies have shown that a variety of chemopreventive plant components affect tumor initiation, promotion, and progression and the main difference, between botanical medicines and synthetic drugs, resides in the presence of complex metabolite mixtures shown by botanical medicine which in turn exert their action on different levels and via different mechanisms. In the present study, we performed an in vitro screening of ethanol extracts from commercial plants in order to investigate potential antitumor activity against human tumor cell lines. Experimental results obtained through a variety of methods and techniques indicated that extracts of I. verum, G. glabra, R. Frangula, and L. usitatissimum present significant reduction in in vitro tumor cell proliferation, suggesting these extracts as possible chemotherapeutical adjuvants for different cancer treatments.

  4. Development of In Vitro Co-Culture Model in Anti-Cancer Drug Development Cascade.

    Science.gov (United States)

    Xu, Ruiling; Richards, Frances M

    2017-01-01

    Tumour microenvironment is recognized as a major determinant of intrinsic resistance to anticancer therapies. In solid tumour types, such as breast cancer, lung cancer and pancreatic cancer, stromal components provide a fibrotic niche, which promotes stemness, EMT, chemo- and radioresistance of tumour. However, this microenvironment is not recapitulated in the conventional cell monoculture or xenografts, hence these in vitro and in vivo preclinical models are unlikely to be predictive of clinical response; which might attribute to the poor predictively of these preclinical drug-screening models. In this review, we summarized recently developed co-culture platforms in various tumour types that incorporate different stromal cell types and/or extracellular matrix (ECM), in the context of investigating potential mechanisms of stroma-mediated chemoresistance and evaluating novel agents and combinations. Some of these platforms will have great utility in the assessment of novel drug combinations and mechanistic understanding of the tumor-stroma interactions. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  5. Screening of selected pesticides for oestrogen receptor activation in vitro

    DEFF Research Database (Denmark)

    Vinggaard, Anne; Breinholt, Vibeke; Larsen, John Christian

    1999-01-01

    Twenty pesticides were tested for their ability to activate the oestrogen receptor in vitro using an,MCF7 cell proliferation assay and a Yeast Oestrogen Screen. The fungicides fenarimol, triadimefon, and triadimenol were identified as weak oestrogen receptor agonists, which at 10 mu M induces a 2.......0, 2.4, and 1.9-fold increase in proliferation of human MCF7 breast cancer cells (E3 clone). The relative proliferation efficiency (RPE) was 43-69%, indicating partial agonism at the oestrogen receptor. Several pesticides did not have any effect oil the proliferation response after 6 days of exposure......, including. chlorpyrifos, diuron, iprodion, linuron, pentachlorphenol, prochloraz, propioconazol, propyzamine, quintozen, tetrachorvinphos and tetradifon. Some pesticides resulted in a negligible proliferation response, which was nor statistically significant under the present experimental conditions...

  6. In vitro screening of potato genotypes for osmotic stress tolerance

    Directory of Open Access Journals (Sweden)

    Gelmesa Dandena

    2017-02-01

    genotypes. Most of the genotypes collected from Ethiopia were found to be susceptible to osmotic stress, except one farmers’ cultivar (Dadafa and two improved varieties (Zemen and Belete. Field evaluation of the tested materials under drought conditions would confirm the capacity of osmotic stress tolerant genotypes to perform well under drought-prone conditions and the potential interest of in vitro evaluation as a pre-screening component in potato breeding programs.

  7. Quantitative high-throughput screen identifies inhibitors of the Schistosoma mansoni redox cascade.

    Directory of Open Access Journals (Sweden)

    Anton Simeonov

    2008-01-01

    Full Text Available Schistosomiasis is a tropical disease associated with high morbidity and mortality, currently affecting over 200 million people worldwide. Praziquantel is the only drug used to treat the disease, and with its increased use the probability of developing drug resistance has grown significantly. The Schistosoma parasites can survive for up to decades in the human host due in part to a unique set of antioxidant enzymes that continuously degrade the reactive oxygen species produced by the host's innate immune response. Two principal components of this defense system have been recently identified in S. mansoni as thioredoxin/glutathione reductase (TGR and peroxiredoxin (Prx and as such these enzymes present attractive new targets for anti-schistosomiasis drug development. Inhibition of TGR/Prx activity was screened in a dual-enzyme format with reducing equivalents being transferred from NADPH to glutathione via a TGR-catalyzed reaction and then to hydrogen peroxide via a Prx-catalyzed step. A fully automated quantitative high-throughput (qHTS experiment was performed against a collection of 71,028 compounds tested as 7- to 15-point concentration series at 5 microL reaction volume in 1536-well plate format. In order to generate a robust data set and to minimize the effect of compound autofluorescence, apparent reaction rates derived from a kinetic read were utilized instead of end-point measurements. Actives identified from the screen, along with previously untested analogues, were subjected to confirmatory experiments using the screening assay and subsequently against the individual targets in secondary assays. Several novel active series were identified which inhibited TGR at a range of potencies, with IC(50s ranging from micromolar to the assay response limit ( approximately 25 nM. This is, to our knowledge, the first report of a large-scale HTS to identify lead compounds for a helminthic disease, and provides a paradigm that can be used to jump

  8. Agar composition affects in vitro screening of biocontrol activity of antagonistic microorganisms.

    Science.gov (United States)

    Bosmans, L; De Bruijn, I; De Mot, R; Rediers, H; Lievens, B

    2016-08-01

    Agar-based screening assays are the method of choice when evaluating antagonistic potential of bacterial biocontrol-candidates against pathogens. We showed that when using the same medium, but different agar compositions, the activity of a bacterial antagonist against Agrobacterium was strongly affected. Consequently, results from in vitro screenings should be interpreted cautiously. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Trypanothione reductase: a target protein for a combined in vitro and in silico screening approach.

    Science.gov (United States)

    Beig, Mathias; Oellien, Frank; Garoff, Linnéa; Noack, Sandra; Krauth-Siegel, R Luise; Selzer, Paul M

    2015-01-01

    With the goal to identify novel trypanothione reductase (TR) inhibitors, we performed a combination of in vitro and in silico screening approaches. Starting from a highly diverse compound set of 2,816 compounds, 21 novel TR inhibiting compounds could be identified in the initial in vitro screening campaign against T. cruzi TR. All 21 in vitro hits were used in a subsequent similarity search-based in silico screening on a database containing 200,000 physically available compounds. The similarity search resulted in a data set containing 1,204 potential TR inhibitors, which was subjected to a second in vitro screening campaign leading to 61 additional active compounds. This corresponds to an approximately 10-fold enrichment compared to the initial pure in vitro screening. In total, 82 novel TR inhibitors with activities down to the nM range could be identified proving the validity of our combined in vitro/in silico approach. Moreover, the four most active compounds, showing IC50 values of <1 μM, were selected for determining the inhibitor constant. In first on parasites assays, three compounds inhibited the proliferation of bloodstream T. brucei cell line 449 with EC50 values down to 2 μM.

  10. In-Vitro Cytotoxicity Screening of Plant Extracts

    National Research Council Canada - National Science Library

    Barrows, Louis

    2002-01-01

    .... Random screening of West African flora for anticancer activity can provide information on the utility of species not recognized by traditional healers for diseases not regularly usually treated at that level.

  11. Phytochemical and in-vitro antimicrobial screening of Sanseviera ...

    African Journals Online (AJOL)

    MICHAEL HORSFALL

    Pharmacognosy & Traditional Medicine, Awka, Anambra state, Nigeria,. KEYWORDS: Sanseviera liberica, Antimicrobial activity, Phytochemical screening, Extracts, Clinical isolates. ABSTRACT: Sanseviera liberica is used in Nigerian folk medicine for the treatment of asthma, abdominal pain, diarrhea, wounds of the foot, ...

  12. In Vitro Screening of Cytotoxic, Antimicrobial and Antioxidant ...

    African Journals Online (AJOL)

    Purpose: To evaluate the in vitro cytotoxic, antioxidant and antimicrobial activities of Clinacanthus nutans extracts and semi-fractions. Method: The plant was subjected to cold solvent extraction to produce petroleum ether, ethyl acetate and methanol crude extracts, followed by isolation using bioassay-guided fractionation.

  13. Preliminary Phytochemical Screening and In vitro Antioxidant Prop ...

    African Journals Online (AJOL)

    The in vitro antioxidant potential of the extracts were also determined using the reducing power and 2, 2-diphenyl-1-picryl-hydrazyl radical (DPPH) scavenging tests respectively. Total phenol content of ... with oxida-tive stress. Keywords: Phytochemicals, total phenol content, reducing power, DPPH, antioxidant, free radicals.

  14. In vitro antimicrobial screening of Aquilaria agallocha roots | Canh ...

    African Journals Online (AJOL)

    ... agallocha roots should have a medicinal uses especially against E. faecium, L. monocytogenes ATCC 7644, B. subtilis DSMZ 1971, C. albicans DSMZ 1386, S. epidermidis DSMZ 20044 and S. aureus ATCC 25923. Keywords: Aquilaria agallocha, Mesir paste, antimicrobial activity, antimicrobial screening, ethanol extract ...

  15. Occurrence and in vitro bioactivity of estrogen, androgen, and glucocorticoid compounds in a nationwide screen of United States stream waters

    Data.gov (United States)

    U.S. Environmental Protection Agency — In vitro bioactivity concentrations and chemical concentrations of estrogens, androgens, and glucocorticoids from a nationwide screen of United States stream water...

  16. In vitro phytochemical and antimicrobial screening of Thymus linearis

    Directory of Open Access Journals (Sweden)

    Attiqa Naz

    2015-03-01

    Full Text Available Extracts from the whole plant of Thymus linearis were extracted with methanol (crude, chloroform, n-hexane, ethyl acetate and butanol and screened for their phytochemical and antimicrobial potentials. Preliminary phytochemical screening of plant extracts manifests the existence of terpenoids, flavonoids, tannins, alkaloids, glycosides and reducing sugars. Fourier transform infrared spectroscopy studies were carried out on various phytochemicals extracted from the extracts of T. linearis which results in the presence of different compounds like amides, aldehydes, carboxylic acid, ethers, alcohol and ketones. All the extracts of T. linearis showed significant antibacterial and antifungal activities when tested against nine bacterial and four fungal strains. It was concluded from this study that extracts of T. linearis have an array of important phytochemicals and significant activities against some of the multidrug resistant bacterial and medically important fungal strains.

  17. In vitro screening of antifungal activity of marine sponge extracts against five phytopathogenic fungi

    OpenAIRE

    El Amraoui, Belkassem; El Wahidi, Majida; Fassouane, Aziz

    2014-01-01

    The aim of our research is the screening of extracts of marine sponges for their antifungal activity against phytopathogenic fungi. The in vitro screening of hydroalcoholic and organic extracts of ten marine sponges from Atlantic coast of Morocco against five phytopathogenic fungi (Fusarium oxysporum f.sp. melonis, Fusarium oxysporum f.sp. radicis-lycopersici, Fusarium oxysporum f.sp. ciceris, Botrytis cinerea and Penicillium digitatum) showed that only two sponges (Haliclona viscosa and Cyna...

  18. In vitro screening for low temperature tolerance of wheat genotypes

    Directory of Open Access Journals (Sweden)

    Kondić-Šipka Ankica

    2006-01-01

    Full Text Available Low temperature (LT tolerance of 12 wheat (T. aestivum L. genotypes was investigated in an in vitro zygotic embryo culture. Isolated embryos were grown on a modified MS medium for two months. Calluses were prepared by gradual decrease of temperatures (17°C-7 days, 5°C-7 days, -5°C-l day, -10°C-1 day and then exposed to LT treatments (-15°C, 24 h and 48 h. After LT treatments, calluses were acclimated by gradual increase of temperatures and grown for another month. Based on the differences in callus fresh weight between the control group and calluses exposed to the LT treatments, levels of LT tolerance of wheat genotypes were determined. The results have shown that out of the 12 wheat cultivars, 6 had a high level of LT tolerance, 5 had a medium and only one spring cultivar (Venera had a low level of LT tolerance. These results were compared with the results of the standard in situ test in a cold chamber. Only partial disagreement of results was determined in two cultivars with the medium level of LT tolerance (NSR-2 and Balkan. It can be concluded that the in vitro test can be successfully used for the separation of genotypes with high and low levels of LT tolerance, but for more precise separation within the medium level of LT tolerance, the test should be improved.

  19. An in vitro screening assay for dental stain cleaning.

    Science.gov (United States)

    Wang, Changxiang; Lucas, Robert; Smith, Anthony J; Cooper, Paul R

    2017-01-09

    The present study aimed to develop an in vitro model for stain removal from natural enamel for the assessment and comparison of oral hygiene products. Bovine teeth (n = 8 per group) were ground/polished to provide flat enamel specimens and ferric-tannate deposits were precipitated onto the enamel surfaces. The ferric-tannate stained enamel specimens were brushed using an in vitro tooth-brushing simulator with slurries containing commercially available toothpaste products, dental abrasive particles, and sodium tripolyphosphate (STP) solutions of different concentrations. The colour of the enamel surfaces was measured using a spectrophotometer before and after stain application as well as after the brushing treatments. Differences in stain removal efficacy were found between the toothpastes categorised as whitening and non-whitening comprising of different types of dental abrasives (hydrated silica and alumina). A mean value of 27% for stain removal was detected for the three non-whitening toothpastes and 59% of stain removal was detected for the three whitening toothpastes after 1000 strokes. Compared with the slurry with Zeodent 113 abrasive alone, the addition of STP provided better performance for stain removal under the same brushing conditions (mean value of 62% for Zeodent 113 abrasive alone and 72% with the addition of 5% (w/w) STP after 1000 strokes). No difference was evident between the STP concentration of 5% (w/w) and 10% (w/w). The ferric-tannate/bovine enamel model reported here provides good stain retention, is rapidly and easily prepared, and is shown to be progressively and reproducibly sensitive to toothbrushing using different toothpastes and surfactant/chelating agent solutions. Importantly, it provides good discrimination between various oral hygiene products. The stain removal assay reported here has considerable potential to enable comparative assessments of different toothpaste types in terms of their cleaning capabilities.

  20. Neurological Condition of Infants Born After In Vitro Fertilization With Preimplantation Genetic Screening

    NARCIS (Netherlands)

    Middelburg, Karin J.; Heineman, Maas J.; Haadsma, Maaike L.; Bos, Arend F.; Kok, Joke H.; Hadders-Algra, Mijna

    Aim of this study was to evaluate the effect of preimplantation genetic screening (PGS) on neurodevelopmental outcome in children. We conducted a prospective follow-up Study of children born to women randomly assigned to in vitro fertilization with or without PGS. Primary outcome was adverse

  1. Neurological Condition of Infants Born After In Vitro Fertilization With Preimplantation Genetic Screening

    NARCIS (Netherlands)

    Middelburg, Karin J.; Jan Heineman, Maas; Haadsma, Maaike L.; Bos, Arend F.; Kok, Joke H.; Hadders-Algra, Mijna

    2010-01-01

    Aim of this study was to evaluate the effect of preimplantation genetic screening (PGS) on neurodevelopmental outcome in children. We conducted a prospective follow-up Study of children born to women randomly assigned to in vitro fertilization with or without PGS. Primary outcome was adverse

  2. Human Vascular Microphysiological System for in vitro Drug Screening

    Science.gov (United States)

    Fernandez, C. E.; Yen, R. W.; Perez, S. M.; Bedell, H. W.; Povsic, T. J.; Reichert, W. M.; Truskey, G. A.

    2016-01-01

    In vitro human tissue engineered human blood vessels (TEBV) that exhibit vasoactivity can be used to test human toxicity of pharmaceutical drug candidates prior to pre-clinical animal studies. TEBVs with 400–800 μM diameters were made by embedding human neonatal dermal fibroblasts or human bone marrow-derived mesenchymal stem cells in dense collagen gel. TEBVs were mechanically strong enough to allow endothelialization and perfusion at physiological shear stresses within 3 hours after fabrication. After 1 week of perfusion, TEBVs exhibited endothelial release of nitric oxide, phenylephrine-induced vasoconstriction, and acetylcholine-induced vasodilation, all of which were maintained up to 5 weeks in culture. Vasodilation was blocked with the addition of the nitric oxide synthase inhibitor L-NG-Nitroarginine methyl ester (L-NAME). TEBVs elicited reversible activation to acute inflammatory stimulation by TNF-α which had a transient effect upon acetylcholine-induced relaxation, and exhibited dose-dependent vasodilation in response to caffeine and theophylline. Treatment of TEBVs with 1 μM lovastatin for three days prior to addition of Tumor necrosis factor – α (TNF-α) blocked the injury response and maintained vasodilation. These results indicate the potential to develop a rapidly-producible, endothelialized TEBV for microphysiological systems capable of producing physiological responses to both pharmaceutical and immunological stimuli. PMID:26888719

  3. IN VITRO ANTIMICROBIAL SCREENING OF AQUILARIA AGALLOCHA ROOTS.

    Science.gov (United States)

    Canlı, Kerem; Yetgin, Ali; Akata, Ilgaz; Altuner, Ergin Murat

    2016-01-01

    It was previously shown that some parts of Aquilaria agallocha, which is commonly known as oud or oodh, such as roots have been used as a traditional medical herbal in different countries. In Turkey A. agallocha is one of the ingredients while preparing famous Mesir paste, which was invented as a medicinal paste and used from the Ottoman period to now at least for 500 years. The identification the in vitro antimicrobial activity of ethanol extract of A. agallocha roots is main purpose of this analysis. By using 17 bacteria and 1 fungi, which include Bacillus, Candida, Enterobacter, Enterococcus, Escherichia, Klebsiella, Listeria, Pseudomonas, Salmonella and Staphylococcus genera, the activity of A. agallocha root extracts were analysed by the help of the disk diffusion method, that is one of the methods commonly used to determine antimicrobial activities. As a result of the study it was observed that ethanol extracts of A. agallocha roots have a clear antimicrobial activity against nearly all microorganism used in the study, but only two bacteria namely E. coli ATCC 25922 and S. typhimurium SL 1344. According to the disk diffusion test results it may be possible to propose that A. agallocha roots should have a medicinal uses especially against E. faecium, L. monocytogenes ATCC 7644, B. subtilis DSMZ 1971, C. albicans DSMZ 1386, S. epidermidis DSMZ 20044 and S. aureus ATCC 25923.

  4. Identification of selective inhibitors of RET and comparison with current clinical candidates through development and validation of a robust screening cascade [version 2; referees: 4 approved

    Directory of Open Access Journals (Sweden)

    Amanda J. Watson

    2016-08-01

    Full Text Available RET (REarranged during Transfection is a receptor tyrosine kinase, which plays pivotal roles in regulating cell survival, differentiation, proliferation, migration and chemotaxis. Activation of RET is a mechanism of oncogenesis in medullary thyroid carcinomas where both germline and sporadic activating somatic mutations are prevalent.   At present, there are no known specific RET inhibitors in clinical development, although many potent inhibitors of RET have been opportunistically identified through selectivity profiling of compounds initially designed to target other tyrosine kinases. Vandetanib and cabozantinib, both multi-kinase inhibitors with RET activity, are approved for use in medullary thyroid carcinoma, but additional pharmacological activities, most notably inhibition of vascular endothelial growth factor - VEGFR2 (KDR, lead to dose-limiting toxicity. The recent identification of RET fusions present in ~1% of lung adenocarcinoma patients has renewed interest in the identification and development of more selective RET inhibitors lacking the toxicities associated with the current treatments.   In an earlier publication [Newton et al, 2016; 1] we reported the discovery of a series of 2-substituted phenol quinazolines as potent and selective RET kinase inhibitors. Here we describe the development of the robust screening cascade which allowed the identification and advancement of this chemical series.  Furthermore we have profiled a panel of RET-active clinical compounds both to validate the cascade and to confirm that none display a RET-selective target profile.

  5. Identification of selective inhibitors of RET and comparison with current clinical candidates through development and validation of a robust screening cascade [version 1; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Amanda J. Watson

    2016-05-01

    Full Text Available RET (REarranged during Transfection is a receptor tyrosine kinase, which plays pivotal roles in regulating cell survival, differentiation, proliferation, migration and chemotaxis. Activation of RET is a mechanism of oncogenesis in medullary thyroid carcinomas where both germline and sporadic activating somatic mutations are prevalent.   At present, there are no known specific RET inhibitors in clinical development, although many potent inhibitors of RET have been opportunistically identified through selectivity profiling of compounds initially designed to target other tyrosine kinases. Vandetanib and cabozantinib, both multi-kinase inhibitors with RET activity, are approved for use in medullary thyroid carcinoma, but additional pharmacological activities, most notably inhibition of vascular endothelial growth factor - VEGFR2 (KDR, lead to dose-limiting toxicity. The recent identification of RET fusions present in ~1% of lung adenocarcinoma patients has renewed interest in the identification and development of more selective RET inhibitors lacking the toxicities associated with the current treatments.   In an earlier publication [Newton et al, 2016; 1] we reported the discovery of a series of 2-substituted phenol quinazolines as potent and selective RET kinase inhibitors. Here we describe the development of the robust screening cascade which allowed the identification and advancement of this chemical series.  Furthermore we have profiled a panel of RET-active clinical compounds both to validate the cascade and to confirm that none display a RET-selective target profile.

  6. In vitro screening of antifungal activity of marine sponge extracts against five phytopathogenic fungi.

    Science.gov (United States)

    El Amraoui, Belkassem; El Wahidi, Majida; Fassouane, Aziz

    2014-01-01

    The aim of our research is the screening of extracts of marine sponges for their antifungal activity against phytopathogenic fungi. The in vitro screening of hydroalcoholic and organic extracts of ten marine sponges from Atlantic coast of Morocco against five phytopathogenic fungi (Fusarium oxysporum f.sp. melonis, Fusarium oxysporum f.sp. radicis-lycopersici, Fusarium oxysporum f.sp. ciceris, Botrytis cinerea and Penicillium digitatum) showed that only two sponges (Haliclona viscosa and Cynachirella tarentina) are active against all phytopathogenic fungi studied.

  7. Screening of cesticidal compounds on a tapeworm hymenolepis nana in vitro

    Science.gov (United States)

    Sen, A. B.; Hawking, F.

    1960-01-01

    A simple and convenient in vitro technique is described for the screening of compounds for action against Hymenolepis nana and probably many other intestinal worms. The results obtained from this test are in broad agreement with the findings of clinical experience and of a small series of in vivo tests. Among the substances tested, the most active ones were oil of chenopodium, dichlorophen, extract of cashew nut (Anacardium occidentale), antimony potassium tartrate, and BIQ 20 [eicosamethylenebis(isoquinolinium iodide)]. PMID:13750047

  8. Exploring the metal phytoremediation potential of three Populus alba L. clones using an in vitro screening.

    Science.gov (United States)

    Di Lonardo, Sara; Capuana, Maurizio; Arnetoli, Miluscia; Gabbrielli, Roberto; Gonnelli, Cristina

    2011-01-01

    This work was planned for providing a useful screening tool for the selection of Populus alba clones suitable for phytoremediation techniques. To this aim, we investigated variation in arsenic, cadmium, copper, and zinc tolerance, accumulation and translocation in three poplar clones through an in vitro screening. Poplars have been widely proposed for phytoremediation, as they are adaptable to grow on contaminated areas and able to accumulate metals. The investigation of possible differences among poplar clones in metal tolerance and accumulation deserves to be deeply studied and exploited for the selection of the more suitable tool for phytoremediation purposes. In vitro multiplied microshoots of a commercial and two autochthonous P. alba clones were subcultured on hormone-free WPM medium for 1 month and then transferred for 2 weeks onto media containing different concentrations of the metals investigated. At the end of the treatments, plantlets were sampled, weighed, and mineralised by wet ashing. Metal concentrations were determined by ICP-OES. For the metal concentration used in the experiments, our clones of P. alba showed variation in metal tolerance, metal accumulation and content. The fast-growing commercial clone, even if rarely showing the highest plant metal concentration, displayed the highest metal content, suggesting biomass production as the key factor in evaluating the phytoextraction capacity of P. alba clones for the metals studied. Data demonstrated that in vitro screening of cuttings represents a valuable way of assessing the ability of different poplar clones to take up, tolerate and survive metal stress.

  9. Electroluminescent TCC, C3dg and fB/Bb epitope assays for profiling complement cascade activation in vitro using an activated complement serum calibration standard.

    Science.gov (United States)

    van Vuuren, B Jansen; Bergseth, G; Mollnes, T E; Shaw, A M

    2014-01-15

    Electroluminescent assays for epitopes on the complement components C3dg, terminal complement complex (TCC) and factor B/Bb (fB/Bb) have been developed with capture and detection antibodies to produce detection limits C3dg=91±9ng/mL, TCC=3±0.1ng/mL and fB=55.7±0.1ng/mL. The assay performance was assessed against a series of zymosan and heat aggregated IgG (HAIgG) in vitro activations of complement using a calibrated activated complement serum (ACS) as calibration standard. The ACS standard was stable within 20% accuracy over a 6-month period with freeze-thaw cycles as required. Differential activation of the complement cascade was observed for TCC showing a pseudo-first order formation half-life of 3.5h after activation with zymosan. The C3dg activation fragment indicates a 10% total activation for both activation agents. The kinetic-epitope analysis for fB indicates that the capture epitope is on the fB/Bb protein fragment which can then become covered by the formation of C3bBb or C3bBbP complexes during the time course of the cascade. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. In vitro and in vivo screening of azole fungicides for antiandrogenic effects

    DEFF Research Database (Denmark)

    Taxvig, Camilla; Vinggaard, Anne; Hass, Ulla

    and antiandrogenic effects both in vitro and in vivo. Two other azole fungicides, tebuconazole and epoxiconazole, have now been investigated for antiandrogenic effects in vitro and in vivo as well. The fungicides were screened in two well-established cell assays, including testing for agonistic and antagonistic...... effects on AR in transfected CHO cells, using an AR reporter gene assay. The compounds were also analyzed for effects on steroidogenesis in H295R cells, a human adrenocorticocarcinoma cell line, used to detect effects on steroid production. In vitro tebuconazole and epoxiconazole proved to be antagonists...... signs of feminization of the male offspring were investigated. Tebuconazole caused an increase in testicular 17alfa-hydroxyprogesterone and progesterone levels, and a decrease in testosterone levels in male fetuses. Epoxiconazole had no effect on any of the mesured hormonelevels. Furthermore...

  11. In vitro screening of azole fungicides for antiandrogenic effects – comparison with in vivo effects

    DEFF Research Database (Denmark)

    -based assays. In vitro prochloraz proved to be an activator of the aryl hydrocarbon receptor (AhR), to inhibit aromatase activity and to possess antiestrogenic and antiandrogenic effects both in vitro and in vivo. Another two azole fungicides, tebuconazole and epoxiconazole, have now been investigated...... in H295R cells, a cell line, which produces a wide range of steroid hormones in measurable quantities, including testosterone, progesterone and estradiol, a property that makes it suitable as a screening assay to detect effects on steroidogenesis. In the in vitro tebuconazole and epoxiconazole showed...... antiandrogenic effects, and in the H295R cell assay, tebuconazole and epoxiconazole were like prochloraz able to inhibit testosterone and estradiol levels and increase progesterone levels. For the in vivo testing, a study was conducted testing the developmental effects on offspring after prenatal exposure...

  12. Screening of selected pesticides for inhibition of CYP19 aromatase activity in vitro

    DEFF Research Database (Denmark)

    Vinggaard, A.M.; Hnida, C.; Breinholt, V.

    2000-01-01

    Many pesticides are able to block or activate the steroid hormone receptors and/or to affect the levels of sex hormones, thereby potentially affecting the development or expression of the male and female reproductive system or both. This emphasizes the relevance of screening pesticides for a wide......, and triadimenol were identified as weak aromatase inhibitors. In conclusion, seven out of 22 tested pesticides turned out to be weak to moderate aromatase inhibitors in vitro, indicating the relevance of elucidating the endocrine effects in vivo of these compounds....

  13. In vitro advanced antimycobacterial screening of isoniazid-related hydrazones, hydrazides and cyanoboranes: part 14.

    Science.gov (United States)

    Maccari, Rosanna; Ottanà, Rosaria; Vigorita, Maria Gabriella

    2005-05-16

    As a part of an ongoing search for new isoniazid-related isonicotinoylhydrazones (ISNEs), 2'-monosubstituted isonicotinohydrazides and cyanoboranes, some analogues belonging to these three series of compounds were further evaluated in an in vitro advanced antimycobacterial screening. The results here reported allowed us to extend their structure-activity relationships. A general correlation emerged between their lipophilicity and effectiveness against intracellular M. tuberculosis. On the whole, the most interesting result of this research was that some hydrazides and ISNEs proved to be more effective antimycobacterial agents than parental isoniazid in a TB-infected macrophage model.

  14. An in vitro model for screening estrogen activity of environmental samples after metabolism

    Energy Technology Data Exchange (ETDEWEB)

    Chahbane, N.; Schramm, K.W. [GSF - Forschungszentrum fuer Umwelt und Gesundheit Neuherberg GmbH, Oberschleissheim (Germany). Inst. fuer Oekologische Chemie; Kettrup, A. [Technische Univ. Muenchen, Freising (Germany). Lehrstuhl fuer Oekologische Chemie

    2004-09-15

    For a few years, yeast estrogen assay (YES) was accepted as a reliable and economic model for screening of environmental estrogens. Though the chemicals directly act with estrogen receptor (ER) can be filtered out by this model, there are still chemicals act with ER only after metabolism and some chemicals eliminate their estrogen activities after metabolism. That is to say, their metabolites exert or have stronger estrogen activities than themselves, which can be called bio-activation. In this case, for the lack of the metabolism enzyme system as human and other animals, only the assay with recombinant yeast cells is insufficient. So, it is necessary to combine the YES with metabolism procedure to evaluate the estrogen activities of these chemicals. The most common method used currently for in vitro metabolic activation in mutagenicity testing and also be applied to the estrogen screening field is S-9 mixture. Also, there is an attempt to develop a chemical model for cytochrome P450 as a bio-mimetic metabolic activation system. All these methods can be used as in vitro models for metabolism. Compare with these models, using whole H4II E cells for metabolism is an alternative and with superiorities. It has the excellence of short experiment period as all other in vitro models, but is much more close to the real surroundings as in vivo. Furthermore, the activity of 7-ethoxyresorufin-O-deethylase (EROD) can be easily measured during the whole incubation period for us to discuss the metabolic activities in a quantitative foundation, not only in qualitative. Methoxychlor is one of the chemicals with bio-activation ability. When directly used in the YES, it shows weak estrogen activity. But a main metabolite of methoxychlor, 2,2-bis (p-hydroxyphenyl) - 1,1,1-trichloroethane (HPTE) is a known estrogen mimic. For the long time using methoxychlor as a pesticide and its clear background, it is an ideal chemical to establish this in vitro system.

  15. PICKLES: the database of pooled in-vitro CRISPR knockout library essentiality screens.

    Science.gov (United States)

    Lenoir, Walter F; Lim, Tassica L; Hart, Traver

    2018-01-04

    The adaptation of CRISPR/Cas9 systems for pooled library genetic knockout screens in mammalian cells has substantially advanced the state of the art in human functional genomics. Screening panels of cell lines for genes whose knockout imposes a significant fitness defect has dramatically expanded our catalog of high-confidence essential genes, and has already proven useful in identifying tumor-specific essential genes for the development of targeted therapies. However, nonexperts currently lack an easy to use way to access this data and to identify whether their genes of interest are essential across different genetic backgrounds. The volume of screening data is expected to grow massively, making the problem more intractable. Here we describe PICKLES, the database of Pooled In vitro CRISPR Knockout Library Essentiality Screens, where end users can display and download raw or normalized essentiality profiles for more that 18 000 protein-coding genes across more than 50 cell lines. An additional data set with 15,000 genes targeted by pooled library shRNA in over 100 cell lines is also included. Researchers can see at a glance the relative fitness defect and tissue specificity of their genes of interest, generate and save figures locally, and download all raw data. The database is available at http://pickles.hart-lab.org. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  16. In vitro and in vivo experimental models for drug screening and development for Chagas disease

    Directory of Open Access Journals (Sweden)

    Alvaro José Romanha

    2010-03-01

    Full Text Available Chagas disease, a neglected illness, affects nearly 12-14 million people in endemic areas of Latin America. Although the occurrence of acute cases sharply has declined due to Southern Cone Initiative efforts to control vector transmission, there still remain serious challenges, including the maintenance of sustainable public policies for Chagas disease control and the urgent need for better drugs to treat chagasic patients. Since the introduction of benznidazole and nifurtimox approximately 40 years ago, many natural and synthetic compounds have been assayed against Trypanosoma cruzi, yet only a few compounds have advanced to clinical trials. This reflects, at least in part, the lack of consensus regarding appropriate in vitro and in vivo screening protocols as well as the lack of biomarkers for treating parasitaemia. The development of more effective drugs requires (i the identification and validation of parasite targets, (ii compounds to be screened against the targets or the whole parasite and (iii a panel of minimum standardised procedures to advance leading compounds to clinical trials. This third aim was the topic of the workshop entitled Experimental Models in Drug Screening and Development for Chagas Disease, held in Rio de Janeiro, Brazil, on the 25th and 26th of November 2008 by the Fiocruz Program for Research and Technological Development on Chagas Disease and Drugs for Neglected Diseases Initiative. During the meeting, the minimum steps, requirements and decision gates for the determination of the efficacy of novel drugs for T. cruzi control were evaluated by interdisciplinary experts and an in vitro and in vivo flowchart was designed to serve as a general and standardised protocol for screening potential drugs for the treatment of Chagas disease.

  17. Discovery of novel inhibitors disrupting HIF-1α/von Hippel–Lindau interaction through shape-based screening and cascade docking

    Directory of Open Access Journals (Sweden)

    Xin Xue

    2016-12-01

    Full Text Available Major research efforts have been devoted to the discovery and development of new chemical entities that could inhibit the protein–protein interaction between HIF-1α and the von Hippel–Lindau protein (pVHL, which serves as the substrate recognition subunit of an E3 ligase and is regarded as a crucial drug target in cancer, chronic anemia, and ischemia. Currently there is only one class of compounds available to interdict the HIF-1α/pVHL interaction, urging the need to discover chemical inhibitors with more diversified structures. We report here a strategy combining shape-based virtual screening and cascade docking to identify new chemical scaffolds for the designing of novel inhibitors. Based on this strategy, nine active hits have been identified and the most active hit, 9 (ZINC13466751, showed comparable activity to pVHL with an IC50 of 2.0 ± 0.14 µM, showing the great potential of utilizing these compounds for further optimization and serving as drug candidates for the inhibition of HIF-1α/von Hippel–Lindau interaction.

  18. IN VITRO SCREENING OF LOCALLY ISOLATED LACTOBACILLUS SPECIES FOR PROBIOTIC PROPERTIES

    Directory of Open Access Journals (Sweden)

    M. ASHRAF, M. ARSHAD, M. SIDDIQUE AND G. MUHAMMAD1

    2009-10-01

    Full Text Available The present study was conducted to determine the probiotic properties of locally isolated lactobacilli in-vitro conditions. For this purpose, intestinal contents (n=20 were collected from crop, gizzard, ileum and caecum of adult healthy chicks and conventional yogurt samples (n=20 were procured from the local market for the isolation of lactobacilli. These samples were mixed homogeneously in sterilized phosphate buffer saline (PBS separately. Samples from both sources were inoculated on deMan Rogosa and Sharpe (MRS agar. L. acidophilus 3, L. rhamnosus and L. salivarius were isolated from intestinal contents, while L. delbrucekii ssp bulgaricus and L. paracasei ssp paracasei 1 were isolated from yogurt samples. These lactobacilli were identified through standard API-50 CHL system and then screened for resistance against bile salt, acidic pH, gastric transit and ability to inhibit pathogens as well as survival under different storage temperatures. Tolerance level was found variable (P<0.05 among all the tested species of lactobacillus. All the tested species, except L. delbrucekii and L. paracasei, showed good survival (P<0.05. All lactobacilli inhibited the growth of E. coli and Staphylococcus aureus, except L. delbrucekii that showed significantly (P<0.05 low antimicrobial effect. The results showed that L. acidophilus 3, L. rhamnosus and L. salivarius fulfilled the criteria of in-vitro screening for probiotic properties.

  19. In vitro affinity screening of protein and peptide binders by megavalent bead surface display.

    Science.gov (United States)

    Diamante, Letizia; Gatti-Lafranconi, Pietro; Schaerli, Yolanda; Hollfelder, Florian

    2013-10-01

    The advent of protein display systems has provided access to tailor-made protein binders by directed evolution. We introduce a new in vitro display system, bead surface display (BeSD), in which a gene is mounted on a bead via strong non-covalent (streptavidin/biotin) interactions and the corresponding protein is displayed via a covalent thioether bond on the DNA. In contrast to previous monovalent or low-copy bead display systems, multiple copies of the DNA and the protein or peptide of interest are displayed in defined quantities (up to 10(6) of each), so that flow cytometry can be used to obtain a measure of binding affinity. The utility of the BeSD in directed evolution is validated by library selections of randomized peptide sequences for binding to the anti-hemagglutinin (HA) antibody that proceed with enrichments in excess of 10(3) and lead to the isolation of high-affinity HA-tags within one round of flow cytometric screening. On-bead K(d) measurements suggest that the selected tags have affinities in the low nanomolar range. In contrast to other display systems (such as ribosome, mRNA and phage display) that are limited to affinity panning selections, BeSD possesses the ability to screen and rank binders by their affinity in vitro, a feature that hitherto has been exclusive to in vivo multivalent cell display systems (such as yeast display).

  20. Contribution to in vitro screening of Egyptian plants for schistosomicidal activity.

    Science.gov (United States)

    Yousif, Fouad; Wassel, Gamila; Boulos, Loutfy; Labib, Therese; Mahmoud, Khaled; El-Hallouty, Salwa; El Bardicy, Samia; Mahmoud, Soheir; Ramzy, Fatem; Gohar, Lamiaa; El-Manawaty, May; El Gendy, Mohamed A M; Fayad, Walid; El-Menshawi, Bassem

    2012-06-01

    This study is a continuation of our previous work in which a bioassay screening of 346 methanol extracts from 281 Egyptian plant species was carried out for in vitro schistosomicidal activity. Another 309 methanol extracts from 278 plant species were subjected to the bioassay screening using the same technique on viable Schistosoma mansoni Sambon (Schistosomatidae) mature worms in specialized culture medium (Roswell Park Memorial Institute medium 1640) in a trial to discover a source for a schistosomiasis drug from Egyptian flora. The methanol plant extracts were tested in vitro against viable S. mansoni mature worms in culture medium. Viability of worms was examined after exposure to 100 μg/ml of the extract in the medium for 24 h. Negative (dimethyl sulfoxide) and positive (praziquantel) controls were simultaneously used. Extracts showing schistosomicidal activity were further subjected to determination of their (Lethal concentration) LC₅₀ and LC₉₀ values. Confirmed in vitro antischistosomal activity was found in 42 extracts. Of these, 14 plant species possessed considerably high antischistosomal activity (LC₅₀ ≤ 15 µg/ml), viz. Callistemon viminalis (Soland. Ex Gaertn) Cheel, C. rigidus R.Br., C. speciosus (Sims.) DC, C. citrinus Stapf, Eucalyptus citriodora Hook, E. rostrata Dehnh., Eugenia edulis Vell, E. javanica Lam syn. Syzygium samarangense (Blume) Merril, Melaleuca leucadendron (L.) L., M. stypheloides Sm. (all belong to Myrtaceae), Cryptostegia grandiflora R.Br. (Asclepiadaceae), Zilla spinosa (L.) Prantl (Cruciferae), Ficus trijuja L. (Moraceae) and Fagonia mollis Delile (Zygophylacae). These species may represent additional natural sources of bioactive material that deserve further investigation for drug discovery against schistosomiasis.

  1. Cheburator software for automatically calculating drug inhibitory concentrations from in vitro screening assays.

    Directory of Open Access Journals (Sweden)

    Dmitry Nevozhay

    Full Text Available In the process of new cancer drug development, as the first step of their assessment, their activities are usually studied in vitro against a panel of cancer cell lines. The results of these in vitro drug screening assays are commonly expressed as inhibitory concentration 50% (IC50: the concentration of the tested agent that inhibits the proliferation of the cancer cell population to 50% of the theoretically possible effect (absolute IC50 or maximum effect practically achieved by the drug (relative IC50. The currently available software for calculating IC50 values requires manual data entry, is time consuming, and is prone to calculation errors. Thus, we have developed open source, free, easy-to-use software for performing standardized data evaluations and automatically calculating the IC50. This software eliminates the laborious and error-prone manual entry of data, substantially reduces the amount of time spent for data analysis. It has been extensively used in our department as the main tool for in vitro data processing during the past several years and can be useful for other research groups working in the area of anticancer drug discovery, either alone or combined with other software packages. The current version of our program, Cheburator, together with sample data, source code, and documentation, is freely available at the following URL: http://www.cheburator.nevozhay.com (it is free for academic use, but a license is required for commercial use.

  2. An in vitro spinal cord injury model to screen neuroregenerative materials.

    Science.gov (United States)

    Weightman, Alan P; Pickard, Mark R; Yang, Ying; Chari, Divya M

    2014-04-01

    Implantable 'structural bridges' based on nanofabricated polymer scaffolds have great promise to aid spinal cord regeneration. Their development (optimal formulations, surface functionalizations, safety, topographical influences and degradation profiles) is heavily reliant on live animal injury models. These have several disadvantages including invasive surgical procedures, ethical issues, high animal usage, technical complexity and expense. In vitro 3-D organotypic slice arrays could offer a solution to overcome these challenges, but their utility for nanomaterials testing is undetermined. We have developed an in vitro model of spinal cord injury that replicates stereotypical cellular responses to neurological injury in vivo, viz. reactive gliosis, microglial infiltration and limited nerve fibre outgrowth. We describe a facile method to safely incorporate aligned, poly-lactic acid nanofibre meshes (±poly-lysine + laminin coating) within injury sites using a lightweight construct. Patterns of nanotopography induced outgrowth/alignment of astrocytes and neurons in the in vitro model were strikingly similar to that induced by comparable materials in related studies in vivo. This highlights the value of our model in providing biologically-relevant readouts of the regeneration-promoting capacity of synthetic bridges within the complex environment of spinal cord lesions. Our approach can serve as a prototype to develop versatile bio-screening systems to identify materials/combinatorial strategies for regenerative medicine, whilst reducing live animal experimentation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. SCREENING APPROACHES FOR METHANE MITIGATING POTENTIAL OF TANNIN-CONTAINING PLANTS UNDER IN VITRO RUMEN ENVIRONMENT

    Directory of Open Access Journals (Sweden)

    A. Jayanegara

    2014-10-01

    Full Text Available The aim of the present study was to conduct univariate, bivariate and multivariate (principalcomponent analysis, PCA approaches in the screening of tannin-containing plants from variouscollection sites for their CH4 mitigating properties. Plant samples were obtained from various collectionsites in different countries, i.e. Indonesia (n = 27 species, Mongolia (n = 14, Switzerland (n = 16 andGermany (n = 3. The plants were incubated in vitro with buffered-rumen fluid at 39oC for 24 h. Totalgas production was recorded as an indicator of feed quality and emission of CH4 was measured. Resultsshowed that, based on bivariate screening, generally, plants possessed low CH4 production had lowquality or low total gas production except Rhus typhina, i.e. 43 ml/200 mg DM. The loading plot of PCAshowed that all phenolic fractions were in the opposite direction with CH4 and total gas production.Plants clustered together in reverse direction to that of CH4 were Bergenia crassifolia root and leaf,Swietenia mahagoni, Clidemia hirta, Peltiphyllum peltatum, Acacia villosa and R. typhina. It wasconluded that, for tannin-containing plants, screenings based on univariate, bivariate and multivariateapproaches in relation to ruminal CH4 emission led to similar results.

  4. In vitro screening of traditional medicines for anti-HIV activity: memorandum from a WHO meeting.

    Science.gov (United States)

    1989-01-01

    Many plant products are being used by patients with acquired immunodeficiency syndrome (AIDS) in some countries without any scientific proof that they possess anti-HIV (human immunodeficiency virus) activity. Traditional healers are now offering their remedies for scientific evaluation, and a few studies provide information on the inhibitory activity against HIV of plants such as Viola yedoensis, Arctium lappa, Epimedium grandiflorum, Glycyrrhiza uralensis and Castanospermum australe. Natural products can be selected for biological screening based on ethnomedical use, random collection or a chemotaxonomic approach (i.e., screening of species of the same botanical family for similar compounds), but the follow-up and selection of plants based on literature leads would seem to be the most cost-effective way of identifying plants with anti-HIV activity. No single in vitro screening methodology for anti-HIV activity is ideal and confirmatory assays in multiple systems are needed to examine completely the potential use of a compound. To promote further research in traditional medicine and AIDS, appropriate institutions will be identified where the different activities for the scientific evaluation of plants and their extracts for possible treatment of AIDS can be carried out. PMID:2633879

  5. Characterization of Apoptosis Signaling Cascades During the Differentiation Process of Human Neural ReNcell VM Progenitor Cells In Vitro.

    Science.gov (United States)

    Jaeger, Alexandra; Fröhlich, Michael; Klum, Susanne; Lantow, Margareta; Viergutz, Torsten; Weiss, Dieter G; Kriehuber, Ralf

    2015-11-01

    Apoptosis is an essential physiological process accompanying the development of the central nervous system and human neurogenesis. However, the time scale and the underlying molecular mechanisms are yet poorly understood. Due to this fact, we investigated the functionality and general inducibility of apoptosis in the human neural ReNcell VM progenitor cell line during differentiation and also after exposure to staurosporine (STS) and ultraviolet B (UVB) irradiation. Transmission light microscopy, flow cytometry, and Western-/Immunoblot analysis were performed to compare proliferating and differentiating, in addition to STS- and UVB-treated cells. In particular, from 24 to 72 h post-initiation of differentiation, G0/G1 cell cycle arrest, increased loss of apoptotic cells, activation of pro-apoptotic BAX, Caspase-3, and cleavage of its substrate PARP were observed during cell differentiation and, to a higher extent, after treatment with STS and UVB. We conclude that redundant or defective cells are eliminated by apoptosis, while otherwise fully differentiated cells were less responsive to apoptosis induction by STS than proliferating cells, likely as a result of reduced APAF-1 expression, and increased levels of BCL-2. These data provide the evidence that apoptotic mechanisms in the neural ReNcell VM progenitor cell line are not only functional, but also inducible by external stimuli like growth factor withdrawal or treatment with STS and UVB, which marks this cell line as a suitable model to investigate apoptosis signaling pathways in respect to the differentiation processes of human neural progenitor cells in vitro.

  6. In vitro cytotoxicity screening of wild plant extracts from Saudi Arabia on human breast adenocarcinoma cells.

    Science.gov (United States)

    Ali, M A; Abul Farah, M; Al-Hemaid, F M; Abou-Tarboush, F M

    2014-05-23

    This study investigated the in vitro anticancer activities of a total of 14 wild angiosperms collected in Saudi Arabia. The cytotoxic activity of each extract was assessed against human breast adenocarcinoma (MCF-7) cell lines by using the MTT assay. Among the plants screened, the potential cytotoxic activity exhibited by the extract of Lavandula dentata (Lamiaceae) was identified, and we analyzed its anticancer potential by testing antiproliferative and apoptotic activity. Our results clearly show that ethanolic extract of L. dentata exhibits promising cytotoxic activity with an IC50 value of 39 μg/mL. Analysis of cell morphological changes, DNA fragmentation and apoptosis (using an Annexin V assay) also confirmed the apoptotic effect of L. dentata extract, and thus, our data call for further investigations to determine the active chemical constituent(s) and their mechanisms of inducing apoptosis.

  7. In vitro screening of snake venom against multidrug-resistant tuberculosis

    Directory of Open Access Journals (Sweden)

    Sujay Kumar Bhunia

    2015-12-01

    Full Text Available The re-emergence of multidrug-resistant tuberculosis (MDR-TB has brought to light the importance of screening effective novel drugs. In the present study, in vitro activities of different snake (Naja naja, Bungarus fasciatus, Daboia russelli russelli, Naja kaouthia venoms have been investigated against clinical isolate of MDR-TB strains. The treatment with all the venoms inhibited the mycobacterial growth for at least a week in common and two of them (Naja naja and Naja kaouthia showed significantly longer inhibition up to two weeks against the MDR-TB strain with single dose and a repetition of those two venoms exhibited inhibition up to more than four weeks.

  8. In vitro screening of nanomedicines through the blood brain barrier: A critical review.

    Science.gov (United States)

    Aparicio-Blanco, Juan; Martín-Sabroso, Cristina; Torres-Suárez, Ana-Isabel

    2016-10-01

    The blood-brain barrier accounts for the high attrition rate of the treatments of most brain disorders, which therefore remain one of the greatest health-care challenges of the twenty first century. Against this background of hindrance to brain delivery, nanomedicine takes advantage of the assembly at the nanoscale of available biomaterials to provide a delivery platform with potential to raising brain levels of either imaging or therapeutic agents. Nevertheless, to prevent later failure due to ineffective drug levels at the target site, researchers have been endeavoring to develop a battery of in vitro screening procedures that can predict earlier in the drug discovery process the ability of these cutting-edge drug delivery platforms to cross the blood-brain barrier for biomedical purposes. This review provides an in-depth analysis of the currently available in vitro blood-brain barrier models (both cell-based and non-cell-based) with the focus on their suitability for understanding the biological brain distribution of forthcoming nanomedicines. The relationship between experimental factors and underlying physiological assumptions that would ultimately lead to a more predictive capacity of their in vivo performance, and those methods already assayed for the evaluation of the brain distribution of nanomedicines are comprehensively discussed. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. In vitro screening of the endocrine disrupting potency of brominated flame retardants and their metabolites

    Energy Technology Data Exchange (ETDEWEB)

    Hamers, T.; Kamstra, J.H. [Inst. for Environmental Studies (IVM), Amsterdam (Netherlands); Sonneveld, E. [BioDetection Systems (BDS), Amsterdam (Netherlands); Murk, A.J. [Wageningen Univ., Toxicology Group, Wageningen (Netherlands); Zegers, B.N.; Boon, J.P. [Royal Netherlands Inst. for Sea Research (NIOZ), Den Burg (Netherlands); Brouwer, A. [Umea Univ., Umea (Sweden)

    2004-09-15

    Substantial evidence is recently becoming available that brominated flame retardants (BFRs) are potential endocrine disruptors. The toxicological profile of BFRs, however, is too incomplete and insufficient to perform human and ecological risk assessment. To fill these gaps, the EU funded research program FIRE was started in December 2002. This program aims at the identification and toxicological characterization of the most potent and environmentally relevant BFRs and their possible risk for human and wildlife health. As part of a hazard identification approach, twenty seven BFRs have been selected within the framework of FIRE for pre-screening their endocrinedisrupting potencies. Selection of test compounds was based on a maximal variation in physicochemical characteristics of BFRs within the test set, allowing the establishment of quantitative structure-activity relationships (QSARs). In addition, environmental relevance (e.g. high production volumes and persistence) and availability for testing were used as selection criteria. BFRs were tested in seven different in vitro bioassays for their potency to interfere via estrogenic, thyroidal, androgenic, progestagenic, and Ah-receptor mediated pathways. Metabolisation rates of BFRs were determined using phenobarbital-induced rat liver microsomes. Finally, the endocrine disrupting potency of the metabolites was determined in the same in vitro bio-assays and compared to the potency of the parent compounds.

  10. In vitro screening of durum wheat against water-stress mediated through polyethylene glycol

    Directory of Open Access Journals (Sweden)

    Nadia Sandra Kacem

    2017-06-01

    Full Text Available Three durum wheat (Triticum durum Desf. genotypes with three levels of drought tolerance were screened in order to evaluate their response to water stress at callus induction and plant regeneration levels. Significant differences were observed among the genotypes, and polyethylene glycol (PEG levels used, and their interactions were however, significant for all the studied characters. Increase in PEG concentration increased the time required for callus initiation and reduced the number of calli frequency of embryogenic structures and number of plants regenerated, showing the adverse effect of PEG on the somatic embryogenesis developmental., under in vitro conditions tested, and Djenah Khetifa was the most tolerant genotype, followed by Oued Zenati and Waha. This pattern was per their drought tolerance behavior under field conditions. Principal component analysis (PCA showed that 95.56% of the total variation was explained by the first two principal components. Biplot analysis allowed the stress-tolerant genotype to be distinguished from the two less tolerant genotypes. Time required for callus initiation was strongly negatively correlated with all other studied traits. These traits can be recommended as suitable selection criteria for screening drought-tolerant genotypes. The selected cells and plants will provide a tool for determining the mechanisms involved in tolerance to water stress.

  11. In vitro bioactivity and phytochemical screening of Suaeda maritima (Dumort): a mangrove associate from Bhitarkanika, India.

    Science.gov (United States)

    Patra, J K; Dhal, N K; Thatoi, H N

    2011-09-01

    To investigate the in vitro antioxidant and antimicrobial activities along with phytochemical screening of organic and aqueous extracts of leaf and stem of Suaeda maritima (Dumort), a mangrove associate from Bhitarkanika of Odisha, India. Antioxidant activity of the crude extracts was evaluated in terms of total antioxidant capacity, total phenol content, ascorbic acid content, DPPH radical scavenging, metal chelating, nitric oxide scavenging, and reducing power etc. The antimicrobial activity of the plant was determined by agar well diffusion method along with MIC and MBC carried out by microdilution techniques against 10 gram positive and gram negative human pathogenic bacteria. The qualitative and quantative phytochemical screening were carried out by standard biochemical assays. Out of the seven antioxidant bioassays, both the leaf and stem extracts were found to posses strong antioxidant properties of 70 % to 92 % for phenol, total antioxidant capacity, DPPH free radical scavenging activity and fairly good ascorbic acid content, metal chelating (1.33 %-22.55 %), reducing power (0.01-0.12) and nitric oxide scavenging (0.84 %-66.99 %) activities. Out of the four extracts evaluated for antimicrobial activity, two leaf extracts such as acetone and ethanol showed promising activity against four pathogenic bacteria and one stem methanol extracts against one pathogenic bacteria when compared with amoxcycillin as standard. The MIC and MBC values of the antimicrobial extracts ranged between 2.5 to 5.0 mg/mL. Screening of phytochemicals showed presence of carbohydrates, protein, tannins, alkaloids and flavonoids in comparatively higher amount than other phytochemicals tested. The present study reveals the presence of potential antioxidants and antimicrobial properties in the plant extract which could be exploited for pharmaceutical application. Copyright © 2011 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  12. Learning Cascading

    CERN Document Server

    Covert, Michael

    2015-01-01

    This book is intended for software developers, system architects and analysts, big data project managers, and data scientists who wish to deploy big data solutions using the Cascading framework. You must have a basic understanding of the big data paradigm and should be familiar with Java development techniques.

  13. Development and Application of In Vitro Models for Screening Drugs and Environmental Chemicals that Predict Toxicity in Animals and Humans

    Science.gov (United States)

    Development and Application of In Vitro Models for Screening Drugs and Environmental Chemicals that Predict Toxicity in Animals and Humans (Presented by James McKim, Ph.D., DABT, Founder and Chief Science Officer, CeeTox) (5/25/2012)

  14. In vitro and in vivo screening of native lactic acid bacteria toward their selection as a probiotic in broiler chickens.

    Science.gov (United States)

    Blajman, Jesica; Gaziano, Cristian; Zbrun, María Virginia; Soto, Lorena; Astesana, Diego; Berisvil, Ayelén; Scharpen, Analía Romero; Signorini, Marcelo; Frizzo, Laureano

    2015-08-01

    Among 360 isolates from the gastrointestinal tract (GIT) of broilers, eleven isolates which showed in vitro probiotic properties were identified and selected for further tests. After the in vitro screening, three strains were chosen for the in vivo study of persistence of fresh cultures and then one strain was selected for the in vivo study of persistence of lyophilized culture. Lyophilized Lactobacillus salivarius DSPV 001P was capable of persisting in broilers during a complete rearing, even 28 days following cessation of administration. L. salivarius DSPV 001P administered to broilers and recovered from GIT was compared by pulsed-field gel electrophoresis (PFGE) to ensure that the same genotype was persistently identified. A combination of in vitro and in vivo screening of native lactic acid bacteria (LAB) described in this study may offer a method for selecting the most suitable strain for potential application as a broiler probiotic supplement. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Identification by virtual screening and in vitro testing of human DOPA decarboxylase inhibitors.

    Directory of Open Access Journals (Sweden)

    Frederick Daidone

    Full Text Available Dopa decarboxylase (DDC, a pyridoxal 5'-phosphate (PLP enzyme responsible for the biosynthesis of dopamine and serotonin, is involved in Parkinson's disease (PD. PD is a neurodegenerative disease mainly due to a progressive loss of dopamine-producing cells in the midbrain. Co-administration of L-Dopa with peripheral DDC inhibitors (carbidopa or benserazide is the most effective symptomatic treatment for PD. Although carbidopa and trihydroxybenzylhydrazine (the in vivo hydrolysis product of benserazide are both powerful irreversible DDC inhibitors, they are not selective because they irreversibly bind to free PLP and PLP-enzymes, thus inducing diverse side effects. Therefore, the main goals of this study were (a to use virtual screening to identify potential human DDC inhibitors and (b to evaluate the reliability of our virtual-screening (VS protocol by experimentally testing the "in vitro" activity of selected molecules. Starting from the crystal structure of the DDC-carbidopa complex, a new VS protocol, integrating pharmacophore searches and molecular docking, was developed. Analysis of 15 selected compounds, obtained by filtering the public ZINC database, yielded two molecules that bind to the active site of human DDC and behave as competitive inhibitors with K(i values ≥10 µM. By performing in silico similarity search on the latter compounds followed by a substructure search using the core of the most active compound we identified several competitive inhibitors of human DDC with K(i values in the low micromolar range, unable to bind free PLP, and predicted to not cross the blood-brain barrier. The most potent inhibitor with a K(i value of 500 nM represents a new lead compound, targeting human DDC, that may be the basis for lead optimization in the development of new DDC inhibitors. To our knowledge, a similar approach has not been reported yet in the field of DDC inhibitors discovery.

  16. Identification by virtual screening and in vitro testing of human DOPA decarboxylase inhibitors.

    Science.gov (United States)

    Daidone, Frederick; Montioli, Riccardo; Paiardini, Alessandro; Cellini, Barbara; Macchiarulo, Antonio; Giardina, Giorgio; Bossa, Francesco; Borri Voltattorni, Carla

    2012-01-01

    Dopa decarboxylase (DDC), a pyridoxal 5'-phosphate (PLP) enzyme responsible for the biosynthesis of dopamine and serotonin, is involved in Parkinson's disease (PD). PD is a neurodegenerative disease mainly due to a progressive loss of dopamine-producing cells in the midbrain. Co-administration of L-Dopa with peripheral DDC inhibitors (carbidopa or benserazide) is the most effective symptomatic treatment for PD. Although carbidopa and trihydroxybenzylhydrazine (the in vivo hydrolysis product of benserazide) are both powerful irreversible DDC inhibitors, they are not selective because they irreversibly bind to free PLP and PLP-enzymes, thus inducing diverse side effects. Therefore, the main goals of this study were (a) to use virtual screening to identify potential human DDC inhibitors and (b) to evaluate the reliability of our virtual-screening (VS) protocol by experimentally testing the "in vitro" activity of selected molecules. Starting from the crystal structure of the DDC-carbidopa complex, a new VS protocol, integrating pharmacophore searches and molecular docking, was developed. Analysis of 15 selected compounds, obtained by filtering the public ZINC database, yielded two molecules that bind to the active site of human DDC and behave as competitive inhibitors with K(i) values ≥10 µM. By performing in silico similarity search on the latter compounds followed by a substructure search using the core of the most active compound we identified several competitive inhibitors of human DDC with K(i) values in the low micromolar range, unable to bind free PLP, and predicted to not cross the blood-brain barrier. The most potent inhibitor with a K(i) value of 500 nM represents a new lead compound, targeting human DDC, that may be the basis for lead optimization in the development of new DDC inhibitors. To our knowledge, a similar approach has not been reported yet in the field of DDC inhibitors discovery.

  17. In vitro bioactivity and phytochemical screening of selected spices used in Mauritian foods

    Directory of Open Access Journals (Sweden)

    Diksa Devi Tacouri

    2013-08-01

    Full Text Available Objective: To investigate the in vitro antioxidant and antimicrobial activities along with phytochemical screening of organic and aqueous extracts of spices used in Mauritian foods. Methods: Antioxidant activity of the crude extracts was evaluated in terms of total antioxidant capacity, total phenol content and total flavonoid content. The antimicrobial activity of the spices was determined by the agar well diffusion method against a gram positive and a gram negative bacteria. The qualitative and quantitative phytochemical screening were carried out by standard biochemical assays. Results: All six spices were found to possess alkaloids, coumarins, flavonoids, saponins, steroids, tannins and phenols. Total phenolic content of the extracts varied between 177 and 1 890 mg GAE/g DW while the total flavonoid content varied between 2.8 and 37.6 mg QE/g DW. All six spices were found to possess strong antioxidant properties as well. Highest value was obtained for cinnamon [(24.930依0.198 µmol Fe2+/g DW] whilst turmeric showed the lowest antioxidant activity [(5.980依0.313 µmol Fe2+/g DW] (P<0.05. All extracts showed promising activity against Staphylococcus aureus and Escherichia coli. The size of the inhibition zones ranged between (11.20依0.23 mm to (26.10依2.09 mm (P<0.05 with turmeric and cinnamon being the most effective against Staphylococcus aureus while garlic was least effective against both E. coli and S. aureus. Conclusions: The present study reveals the presence of potential antioxidant and antimicrobial properties in the extracts of the spices which could be further exploited.

  18. Expenditure Cascades

    OpenAIRE

    Frank, R; Levine, A.; Dijk, O.

    2014-01-01

    Prevailing economic models of consumer behavior completely ignore the well-documented link between context and evaluation. We propose and test a theory that explicitly incorporates this link. Changes in one group's spending shift the frame of reference that defines consumption standards for others just below them on the income scale, giving rise to expenditure cascades. Our model, a descendant of James Duesenberry's relative income hypothesis, predicts the observed ways in which individual sa...

  19. Towards the development of lipid multilayer microarrays for dose dependent in vitro delivery and screening

    Science.gov (United States)

    Kusi-Appiah, Aubrey Emmanuel

    Screening for effects of small molecules on cells grown in culture is a well-established method for drug discovery and testing, and faster throughput at lower cost is needed especially for lipophilic materials. Small-molecule arrays present a promising approach. However, it has been a challenge to use them to obtain quantitative surface based dose-response curves in vitro, especially for lipophilic compounds. This thesis first introduces a simple novel method of surface-mediated delivery of drugs to cells from a microarray of phospholipid multilayers (layers thicker than a bilayer) encapsulating small molecules. The capability of controlling the dosage of the lipophilic molecules delivered to cells using the lipid multilayer microarray assay is further demonstrated using the nanointaglio printing method. This control enabled the variation of the volumes of surface supported lipid micro- and nanostructure arrays fabricated with nanointaglio. The volumes of the lipophilic drug-containing nanostructures were determined using a fluorescence microscope calibrated by atomic-force microscopy. The surface supported lipid volume information was used to obtain EC-50 values for the response of HeLa cells to treatment with three FDA-approved lipophilic anticancer drugs, docetaxel, imiquimod and triethylenemelamine, which were found to be significantly different from neat lipid controls. Features with sub-cellular lateral dimensions were found to be necessary to obtain normal cell adhesion with HeLa cells. Comparison of the microarray data to dose-response curves for the same drugs delivered liposomally from solution revealed quantitative differences in the efficacy values, which may be explained in terms of cell-adhesion playing a more important role in the surface-based assay. Finally, solution encapsulation was done for a library of hydrophilic silicon nanocrystals in order to set a solution standard for comparison with future surface supported delivery of the library. The

  20. In vitro screening of probiotic lactic acid bacteria and prebiotic glucooligosaccharides to select effective synbiotics.

    Science.gov (United States)

    Grimoud, Julien; Durand, Henri; Courtin, Céline; Monsan, Pierre; Ouarné, Françoise; Theodorou, Vassilia; Roques, Christine

    2010-10-01

    Probiotics and prebiotics have been demonstrated to positively modulate the intestinal microflora and could promote host health. Although some studies have been performed on combinations of probiotics and prebiotics, constituting synbiotics, results on the synergistic effects tend to be discordant in the published works. The first aim of our study was to screen some lactic acid bacteria on the basis of probiotic characteristics (resistance to intestinal conditions, inhibition of pathogenic strains). Bifidobacterium was the most resistant genus whereas Lactobacillus farciminis was strongly inhibited. The inhibitory effect on pathogen growth was strain dependent but lactobacilli were the most effective, especially L. farciminis. The second aim of the work was to select glucooligosaccharides for their ability to support the growth of the probiotics tested. We demonstrated the selective fermentability of oligodextran and oligoalternan by probiotic bacteria, especially the bifidobacteria, for shorter degrees of polymerisation and absence of metabolism by pathogenic bacteria. Thus, the observed characteristics confer potential prebiotic properties on these glucooligosaccharides, to be further confirmed in vivo, and suggest some possible applications in synbiotic combinations with the selected probiotics. Furthermore, the distinctive patterns of the different genera suggest a combination of lactobacilli and bifidobacteria with complementary probiotic effects in addition to the prebiotic ones. These associations should be further evaluated for their synbiotic effects through in vitro and in vivo models. Copyright © 2010 Elsevier Ltd. All rights reserved.

  1. A multipumping flow system for in vitro screening of peroxynitrite scavengers.

    Science.gov (United States)

    Ribeiro, Marta F T; Dias, Ana C B; Santos, João L M; Fernandes, Eduarda; Lima, José L F C; Zagatto, Elias A G

    2007-09-01

    Peroxynitrite anion is a reactive nitrogen species formed in vivo by the rapid, controlled diffusion reaction between nitric oxide and superoxide radicals. By reacting with several biological molecules, peroxynitrite may cause important cellular and tissue deleterious effects, which have been associated with many diseases. In this work, an automated flow-based procedure for the in vitro generation of peroxynitrite and subsequent screening of the scavenging activity of selected compounds is developed. This procedure involves a multipumping flow system (MPFS) and exploits the ability of compounds such as lipoic acid, dihydrolipoic acid, cysteine, reduced glutathione, oxidized glutathione, sulindac, and sulindac sulfone to inhibit the chemiluminescent reaction of luminol with peroxynitrite under physiological simulated conditions. Peroxynitrite was generated in the MPFS by the online reaction of acidified hydrogen peroxide with nitrite, followed by a subsequent stabilization by merging with a sodium hydroxide solution to rapidly quench the developing reaction. The pulsed flow and the timed synchronized insertion of sample and reagent solutions provided by the MPFS ensure the establishment of the reaction zone only inside the flow cell, thus allowing maximum chemiluminescence emission detection. The results obtained for the assayed compounds show that, with the exception of oxidized glutathione, all are highly potent scavengers of peroxynitrite at the studied concentrations.

  2. Synthesis, characterization and in vitro anti-invasive activity screening of polyphenolic and heterocyclic compounds.

    Science.gov (United States)

    Parmar, Virinder S; Sharma, Nawal K; Husain, Mofazzal; Watterson, Arthur C; Kumar, Jayant; Samuelson, Lynne A; Cholli, Ashok L; Prasad, Ashok K; Kumar, Ajay; Malhotra, Sanjay; Kumar, Naresh; Jha, Amitabh; Singh, Amarjit; Singh, Ishwar; Himanshu; Vats, Archana; Shakil, Najam A; Trikha, Smriti; Mukherjee, Shubasish; Sharma, Sunil K; Singh, Sanjay K; Kumar, Ajay; Jha, Hriday N; Olsen, Carl E; Stove, Christophe P; Bracke, Marc E; Mareel, Marc M

    2003-03-20

    Invasion is the hallmark of malignant tumors, and is responsible for the bad prognosis of the untreated cancer patients. The search for anti-invasive treatments led us to screen compounds of different classes for their effect in an assay for invasion. Thirty-nine new compounds synthesized in the present study along with 56 already reported compounds belonging mainly to the classes of lactones, pyrazoles, isoxazoles, coumarins, desoxybenzoins, aromatic ketones, chalcones, chromans, isoflavanones have been tested against organotypic confronting cultures of invasive human MCF-7/6 mammary carcinoma cells with embryonic chick heart fragments in vitro. Three of them (a pyrazole derivative, an isoxazolylcoumarin and a prenylated desoxybenzoin) inhibited invasion at concentrations as low as 1 microM; instead of occupying and replacing the heart tissue within 8 days, the MCF-7/6 cells grew around the heart fragments and left it intact, when treated with these compounds. At the anti-invasive concentration of 1 microM, the three compounds did not affect the growth of the MCF-7/6 cells, as shown in the sulforhodamine B assay. Aggregate formation on agar was not stimulated by any of the three anti-invasive compounds, making an effect on the E-cadherin/catenin complex improbable. This is an invasion suppressor that can be activated in MCF-7/6 cells by a number of other molecules. Our data indicate that some polyphenolic and heterocyclic compounds are anti-invasive without being cytotoxic for the cancer cells.

  3. Characterization and in vitro immunomodulatory screening of fructo-oligosaccharides of Asparagus racemosus Willd.

    Science.gov (United States)

    Thakur, Mayank; Connellan, Paul; Deseo, Myrna A; Morris, Carol; Praznik, Werner; Loeppert, Renate; Dixit, V K

    2012-01-01

    Asparagus racemosus Linn. (Fam. Liliaceae) is an ethno-pharmacologically acclaimed Ayurvedic medicinal plant. In the present study, aqueous extract of A. racemosus (ARC) was fractionated and screened for the polysaccharide fraction (ARP). The characterization was done by enzymatic, Size Exclusion, gas chromatography with flame ionization detector (GC-FID), high pressure anion exchange chromatography (HPAEC) and thin layer chromatographic analyses. Phyto-chemical evaluation confirmed the presence of 26.7% of 2→1 linked fructo-oligosaccharides (FOS). They have a degree of polymerization (DP) of nearly 7-8. Cytotoxicity evaluation on P388 cell lines was consistent with low cytotoxicity of the extracts. In vitro Natural Killer (NK) cell activity was evaluated using human peripheral blood mononuclear cells (PBMC) isolated from whole blood on a ficoll-hypaque density gradient. K562 a myeloid leukemia cell line, were used as target cells. ARC, tested over the range 0.2-50 μg/ml, showed a dose-related stimulation of NK cell activity with a peak increase of 16.9±4.4% at 5.6 μg/ml. However, ARP demonstrated a higher stimulatory activity of 51.8±1.2% at 25 μg/ml. The results indicate that the FOS from A. racemosus potentiates the NK cell activity and this could be an important mechanism underpinning the 'Rasayana' properties of this plant. Copyright © 2011 Elsevier B.V. All rights reserved.

  4. Phytochemical screening and in vitro bioactivities of the extracts of aerial part of Boerhavia diffusa Linn.

    Science.gov (United States)

    Apu, Apurba Sarker; Liza, Mahmuda Sultana; Jamaluddin, A T M; Howlader, Md Amran; Saha, Repon Kumer; Rizwan, Farhana; Nasrin, Nishat

    2012-09-01

    To investigate the bioactivities of crude n-hexane, ethyl acetate and methanol extracts of aerial part of Boerhavia diffusa Linn. (B. diffusa) and its phytochemical analysis. The identification of phytoconstituents and assay of antioxidant, thrombolytic, cytotoxic, antimicrobial activities were conducted using specific standard in vitro procedures. The results showed that the plant extracts were a rich source of phytoconstituents. Methanol extract showed higher antioxidant, thrombolytic activity and less cytotoxic activity than those of n-hexane and ethyl acetate extracts of B. diffusa. Among the bioactivities, antioxidant activity was the most notable compared to the positive control and thus could be a potential rich source of natural antioxidant. In case of antimicrobial screening, crude extracts of the plant showed remarkable antibacterial activity against tested microorganisms. All the extracts showed significant inhibitory activity against Candida albicuns, at a concentration of 1000 µg/disc. The present findings suggest that, the plant widely available in Bangladesh, could be a prominent source of medicinally important natural compounds.

  5. In vitro antioxidant activity and phytochemical screening of Garhwal Himalaya medicinal plants

    Directory of Open Access Journals (Sweden)

    Subhash Chandra

    2016-08-01

    Full Text Available Bistorta macrophylla, B.vaccinifolia and Persicaria polystachya are used for the treatment and prevention of many ailments including tuberculosis, inflammation, pyretic, fever, flue, lungs disorders, diarrhea, vomiting, arthritis, gout, kidney stones or hyperacidity and hypertension. This study was aimed to evaluate the possible in vitro antioxidant activity and phytochemical screening of B. macrophylla, B.vaccinifolia and P. polystachya. The results of antioxidant activity study of B. macrophylla showed maximum activity in the methanolic extracts at different concentration of 20, 40, 60, 80 and 100 µg/ml. The percent inhibition of writhing response by the extract was 36.18%, 44.72%, 59.21%. 67.08% and 83.39% respectively. In the present work a potent anti-oxidant activity of methanolic extract of the whole plants of B. macrophylla were demonstrated, validating the ethno pharmacological claims. These experimental findings would further establish the scientific basis of the traditional uses of the plant in the management of different conditions as well as control of different disease.

  6. Bacterial and fungal keratitis in Upper Egypt: in vitro screening of enzymes, toxins and antifungal activity.

    Science.gov (United States)

    Gharamah, Abdullah A; Moharram, Ahmed M; Ismail, Mady A; Al-Hussaini, Ashraf K

    2014-02-01

    This work was conducted to study the ability of bacterial and fungal isolates from keratitis cases in Upper Egypt to produce enzymes, toxins, and to test the isolated fungal species sensitivity to some therapeutic agents. One hundred and fifteen patients clinically diagnosed to have microbial keratitis were investigated. From these cases, 37 bacterial isolates and 25 fungal isolates were screened for their ability to produce extra-cellular enzymes in solid media. In addition, the ability of fungal isolates to produce mycotoxins and their sensitivity to 4 antifungal agents were tested. Protease, lipase, hemolysins, urease, phosphatase, and catalase were detected respectively in 48.65%, 37.84%, 59.46%, 43.24%, 67.57%, and 100% out of 37 bacterial isolates tested. Out of 25 fungal isolates tested during the present study, 80% were positive for protease, 84% for lipase and urease, 28% for blood hemolysis, and 100% for phosphatase and catalase enzymes. Thirteen fungal isolates were able to produce detectable amounts of 7 mycotoxins in culture medium (aflatoxins (B1, B2, G1, and G2), sterigmatocystin, fumagillin, diacetoxyscirpenol, zearalenone, T-2 toxin, and trichodermin). Among the antifungal agents tested in this study, terbinafine showed the highest effect against most isolates in vitro. In conclusion, the ability of bacterial and fungal isolates to produce extracellular enzymes and toxins may be aid in the invasion and destruction of eye tissues, which, in turn, lead to vision loss.

  7. Bacterial and fungal keratitis in Upper Egypt: In vitro screening of enzymes, toxins and antifungal activity

    Directory of Open Access Journals (Sweden)

    Abdullah A Gharamah

    2014-01-01

    Full Text Available Purpose: This work was conducted to study the ability of bacterial and fungal isolates from keratitis cases in Upper Egypt to produce enzymes, toxins, and to test the isolated fungal species sensitivity to some therapeutic agents. Materials and Methods: One hundred and fifteen patients clinically diagnosed to have microbial keratitis were investigated. From these cases, 37 bacterial isolates and 25 fungal isolates were screened for their ability to produce extra-cellular enzymes in solid media. In addition, the ability of fungal isolates to produce mycotoxins and their sensitivity to 4 antifungal agents were tested. Results: Protease, lipase, hemolysins, urease, phosphatase, and catalase were detected respectively in 48.65%, 37.84%, 59.46%, 43.24%, 67.57%, and 100% out of 37 bacterial isolates tested. Out of 25 fungal isolates tested during the present study, 80% were positive for protease, 84% for lipase and urease, 28% for blood hemolysis, and 100% for phosphatase and catalase enzymes. Thirteen fungal isolates were able to produce detectable amounts of 7 mycotoxins in culture medium (aflatoxins (B1, B2, G1, and G2, sterigmatocystin, fumagillin, diacetoxyscirpenol, zearalenone, T-2 toxin, and trichodermin. Among the antifungal agents tested in this study, terbinafine showed the highest effect against most isolates in vitro. Conclusion: In conclusion, the ability of bacterial and fungal isolates to produce extracellular enzymes and toxins may be aid in the invasion and destruction of eye tissues, which, in turn, lead to vision loss.

  8. Screening, Characterization and In Vitro Evaluation of Probiotic Properties Among Lactic Acid Bacteria Through Comparative Analysis.

    Science.gov (United States)

    Devi, Sundru Manjulata; Archer, Ann Catherine; Halami, Prakash M

    2015-09-01

    The present work aimed to identify probiotic bacteria from healthy human infant faecal and dairy samples. Subsequently, an assay was developed to evaluate the probiotic properties using comparative genetic approach for marker genes involved in adhesion to the intestinal epithelial layer. Several in vitro properties including tolerance to biological barriers (such as acid and bile), antimicrobial spectrum, resistance to simulated digestive fluids and cellular hydrophobicity were assessed. The potential probiotic cultures were rapidly characterized by morphological, physiological and molecular-based methods [such as RFLP, ITS, RAPD and (GTG)5]. Further analysis by 16S rDNA sequencing revealed that the selected isolates belong to Lactobacillus, Pediococcus and Enterococcus species. Two cultures of non-lactic, non-pathogenic Staphylococcus spp. were also isolated. The native isolates were able to survive under acidic, bile and simulated intestinal conditions. In addition, these cultures inhibited the growth of tested bacterial pathogens. Further, no correlation was observed between hydrophobicity and adhesion ability. Sequencing of probiotic marker genes such as bile salt hydrolase (bsh), fibronectin-binding protein (fbp) and mucin-binding protein (mub) for selected isolates revealed nucleotide variation. The probiotic binding domains were detected by several bioinformatic tools. The approach used in the study enabled the identification of potential probiotic domains responsible for adhesion of bacteria to intestinal epithelial layer, which may further assist in screening of novel probiotic bacteria. The rapid detection of binding domains will help in revealing the beneficial properties of the probiotic cultures. Further, studies will be performed to develop a novel probiotic product which will contribute in food and feed industry.

  9. In vitro biological screening of the anticholinesterase and antiproliferative activities of medicinal plants belonging to Annonaceae

    Energy Technology Data Exchange (ETDEWEB)

    Formagio, A.S.N.; Vieira, M.C. [Faculdade de Ciências Agrárias, Universidade Federal da Grande Dourados, Dourados, MS (Brazil); Volobuff, C.R.F.; Silva, M.S. [Faculdade de Ciências Biológicas e Ambientais, Universidade Federal da Grande Dourados, Dourados, MS (Brazil); Matos, A.I. [Faculdade de Ciências, Universidade de Lisboa, Lisboa (Portugal); Cardoso, C.A.L. [Curso de Química, Universidade Estadual do Mato Grosso do Sul, Dourados, MS (Brazil); Foglio, M.A.; Carvalho, J.E. [Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas, Universidade Estadual de Campinas, Campinas, SP (Brazil)

    2015-02-13

    The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI{sub 50}) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI{sub 50} values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs.

  10. In vitro biological screening of the anticholinesterase and antiproliferative activities of medicinal plants belonging to Annonaceae.

    Science.gov (United States)

    Formagio, A S N; Vieira, M C; Volobuff, C R F; Silva, M S; Matos, A I; Cardoso, C A L; Foglio, M A; Carvalho, J E

    2015-04-01

    The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI50) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI50 values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs.

  11. Newborn Congenital Cytomegalovirus Screening Based on Clinical Manifestations and Evaluation of DNA-based Assays for In Vitro Diagnostics.

    Science.gov (United States)

    Fujii, Tomoyuki; Oka, Akira; Morioka, Ichiro; Moriuchi, Hiroyuki; Koyano, Shin; Yamada, Hideto; Saito, Shigeru; Sameshima, Hiroshi; Nagamatsu, Takeshi; Tsuchida, Shinya; Inoue, Naoki

    2017-10-01

    To establish a strategy for congenital cytomegalovirus (cCMV) screening and to establish confirmatory assays approved as in vitro diagnostics by the regulatory authorities, we evaluated the clinical risks and performance of diagnostic assays developed by commercial companies, since cCMV infection has significant clinical consequences. Newborns with clinical manifestations considered to be consequences of cCMV infection (n = 575) were screened for the presence of cytomegalovirus (CMV) DNA in urine specimens collected onto filter paper placed in their diapers using the polymerase chain reaction-based assay reported previously. Liquid urine specimens were obtained from all of 20 CMV-positive newborns and 107 of the CMV-negative newborns identified in the screening. We used these 127 specimens, as well as 12 from cCMV cases identified in a previous study and 41 from healthy newborns, to compare the performance of 2 commercial assays and 1 in-house assay. The risk-based screening allowed the identification of cCMV cases at least 10-fold more efficiently than our previous universal screening, although there appears to be a limit to the identification of asymptomatically infected newborns. Although CMV-specific IgM during pregnancy was found frequently in mothers of cCMV newborns, CMV-IgM alone is not an effective diagnostic marker. The urine-filter-based assay and the 3 diagnostic assays yielded identical results. Although risk-based and universal newborn screening strategies for cCMV infection each have their respective advantages and disadvantages, urine-filter-based assay followed by confirmatory in vitro diagnostics assays is able to identify cCMV cases efficiently.

  12. Building a Tiered Approach to In Vitro Predictive Toxicity Screening: A Focus on Assays with In Vivo Relevance

    Science.gov (United States)

    McKim, James M

    2010-01-01

    One of the greatest challenges facing the pharmaceutical industry today is the failure of promising new drug candidates due to unanticipated adverse effects discovered during preclinical animal safety studies and clinical trials. Late stage attrition increases the time required to bring a new drug to market, inflates development costs, and represents a major source of inefficiency in the drug discovery/development process. It is generally recognized that early evaluation of new drug candidates is necessary to improve the process. Building in vitro data sets that can accurately predict adverse effects in vivo would allow compounds with high risk profiles to be deprioritized, while those that possess the requisite drug attributes and a lower risk profile are brought forward. In vitro cytotoxicity assays have been used for decades as a tool to understand hypotheses driven questions regarding mechanisms of toxicity. However, when used in a prospective manner, they have not been highly predictive of in vivo toxicity. Therefore, the issue may not be how to collect in vitro toxicity data, but rather how to translate in vitro toxicity data into meaningful in vivo effects. This review will focus on the development of an in vitro toxicity screening strategy that is based on a tiered approach to data collection combined with data interpretation. PMID:20053163

  13. IspE inhibitors identified by a combination of in silico and in vitro high-throughput screening.

    Directory of Open Access Journals (Sweden)

    Naomi Tidten-Luksch

    Full Text Available CDP-ME kinase (IspE contributes to the non-mevalonate or deoxy-xylulose phosphate (DOXP pathway for isoprenoid precursor biosynthesis found in many species of bacteria and apicomplexan parasites. IspE has been shown to be essential by genetic methods and since it is absent from humans it constitutes a promising target for antimicrobial drug development. Using in silico screening directed against the substrate binding site and in vitro high-throughput screening directed against both, the substrate and co-factor binding sites, non-substrate-like IspE inhibitors have been discovered and structure-activity relationships were derived. The best inhibitors in each series have high ligand efficiencies and favourable physico-chemical properties rendering them promising starting points for drug discovery. Putative binding modes of the ligands were suggested which are consistent with established structure-activity relationships. The applied screening methods were complementary in discovering hit compounds, and a comparison of both approaches highlights their strengths and weaknesses. It is noteworthy that compounds identified by virtual screening methods provided the controls for the biochemical screens.

  14. Using CRISPR/Cas9-Mediated GLA Gene Knockout as an In Vitro Drug Screening Model for Fabry Disease.

    Science.gov (United States)

    Song, Hui-Yung; Chiang, Huai-Chih; Tseng, Wei-Lien; Wu, Ping; Chien, Chian-Shiu; Leu, Hsin-Bang; Yang, Yi-Ping; Wang, Mong-Lien; Jong, Yuh-Jyh; Chen, Chung-Hsuan; Yu, Wen-Chung; Chiou, Shih-Hwa

    2016-12-13

    The CRISPR/Cas9 Genome-editing system has revealed promising potential for generating gene mutation, deletion, and correction in human cells. Application of this powerful tool in Fabry disease (FD), however, still needs to be explored. Enzyme replacement therapy (ERT), a regular administration of recombinant human α Gal A (rhα-GLA), is a currently available and effective treatment to clear the accumulated Gb3 in FD patients. However, the short half-life of rhα-GLA in human body limits its application. Moreover, lack of an appropriate in vitro disease model restricted the high-throughput screening of drugs for improving ERT efficacy. Therefore, it is worth establishing a large-expanded in vitro FD model for screening potential candidates, which can enhance and prolong ERT potency. Using CRISPR/Cas9-mediated gene knockout of GLA in HEK-293T cells, we generated GLA-null cells to investigate rhα-GLA cellular pharmacokinetics. The half-life of administrated rhα-GLA was around 24 h in GLA-null cells; co-administration of proteasome inhibitor MG132 and rhα-GLA significantly restored the GLA enzyme activity by two-fold compared with rhα-GLA alone. Furthermore, co-treatment of rhα-GLA/MG132 in patient-derived fibroblasts increased Gb3 clearance by 30%, compared with rhα-GLA treatment alone. Collectively, the CRISPR/Cas9-mediated GLA-knockout HEK-293T cells provide an in vitro FD model for evaluating the intracellular pharmacokinetics of the rhα-GLA as well as for screening candidates to prolong rhα-GLA potency. Using this model, we demonstrated that MG132 prolongs rhα-GLA half-life and enhanced Gb3 clearance, shedding light on the direction of enhancing ERT efficacy in FD treatment.

  15. In vitro screening for protein tyrosine phosphatase 1B and dipeptidyl peptidase IV inhibitors from selected Nigerian medicinal plants

    OpenAIRE

    Saidu, Yusuf; Muhammad, Suleiman Alhaji; Abbas, Abdullahi Yahaya; Onu, Andrew; Tsado, Ibrahim Mohammed; Muhammad, Luba

    2016-01-01

    Background/Aim: Protein tyrosine phosphatase 1B (PTP 1B) and dipeptidyl peptidase IV (DPP IV) have been identified as one of the drug targets for the treatment of Type-2 diabetes. This study was designed to screen for PTP 1B and DPP-IV inhibitors from some Nigerian medicinal plants. Materials and Methods: PTP 1B and DPP-IV drug discovery kits from Enzo Life Sciences were used to investigate in vitro inhibitory effect of crude methanolic extract of 10 plants; Mangifera indica, Moringa oleifera...

  16. Development of a movement-based in vitro screening assay for the identification of new anti-cestodal compounds.

    Directory of Open Access Journals (Sweden)

    Dominic Ritler

    2017-05-01

    Full Text Available Intestinal cestodes are infecting millions of people and livestock worldwide, but treatment is mainly based on one drug: praziquantel. The identification of new anti-cestodal compounds is hampered by the lack of suitable screening assays. It is difficult, or even impossible, to evaluate drugs against adult cestodes in vitro due to the fact that these parasites cannot be cultured in microwell plates, and adult and larval stages in most cases represent different organisms in terms of size, morphology, and metabolic requirements. We here present an in vitro-drug screening assay based on Echinococcus multilocularis protoscoleces, which represent precursors of the scolex (hence the anterior part of the adult tapeworm. This movement-based assay can serve as a model for an adult cestode screen. Protoscoleces are produced in large numbers in Mongolian gerbils and mice, their movement is measured and quantified by image analysis, and active compounds are directly assessed in terms of morphological effects. The use of the 384-well format minimizes the amount of parasites and compounds needed and allows rapid screening of a large number of chemicals. Standard drugs showed the expected dose-dependent effect on movement and morphology of the protoscoleces. Interestingly, praziquantel inhibited movement only partially within 12 h of treatment (at concentrations as high as 100 ppm and did thus not act parasiticidal, which was also confirmed by trypan blue staining. Enantiomers of praziquantel showed a clear difference in their minimal inhibitory concentration in the motility assay and (R-(--praziquantel was 185 times more active than (S-(--praziquantel. One compound named MMV665807, which was obtained from the open access MMV (Medicines for Malaria Venture Malaria box, strongly impaired motility and viability of protoscoleces. Corresponding morphological alterations were visualized by scanning electron microscopy, and demonstrated that this compound exhibits a

  17. Generation of peanut drought tolerant plants by pingyangmycin-mediated in vitro mutagenesis and hydroxyproline-resistance screening.

    Science.gov (United States)

    Sui, Jiongming; Wang, Ya; Wang, Peng; Qiao, Lixian; Sun, Shimeng; Hu, Xiaohui; Chen, Jing; Wang, Jingshan

    2015-01-01

    In order to enlarge the potential resources of drought-tolerant peanuts, we conducted in vitro mutagenesis with Pingyangmycin (PYM) as the mutagen as well as directed screening on a medium supplemented with Hydroxyproline (HYP). After being extracted from mature seeds (cv. Huayu 20), the embryonic leaflets were cultured on somatic embryogenesis-induction medium with 4 mg/L PYM and the generated embryos were successively transferred to a germination medium with 4 and then 8 mmol/L HYP to screen HYP-tolerant plantlets. After that, these plantlets were grafted and transplanted to the experimental field. In the next generation, all seeds were sown in the field, and phenotype variation and trait segregation can be observed in most of the offspring (M2 generation). The M3 generation individuals were subjected to drought stress at the seedling stages. The activities of SOD and POD were substantially increased in eight offspring of 11 HYP-tolerant, regenerated plants than in their mutagenic parents. To determine the correlation between mutant phenotypes and genomic modification, we carried out a comparison of the DNA polymorphisms between the mutagenic parents and 13 M3 generation individuals from different HYP-tolerant, regenerated plants with SSR primers. Results showed that most mutants and parent plants had signs of polymorphisms. Under drought stress, some M3 generation individuals of 10 original HYP-tolerant, regenerated plants produced more pods than the mutagenic parent; twenty individuals among them produced >60 g pods/plant. M4-generation seeds were tested for quality characteristics by Near Infrared Spectroscopy (NIS) and nine individuals with higher protein content (>30%) and 21 individuals with higher oil content (>58%) were screened. We concluded that the use of PYM-based in vitro mutagenesis in combination with directed screening with HYP is effective for the creation of potential drought-tolerant mutants of peanut.

  18. Generation of peanut drought tolerant plants by pingyangmycin-mediated in vitro mutagenesis and hydroxyproline-resistance screening.

    Directory of Open Access Journals (Sweden)

    Jiongming Sui

    Full Text Available In order to enlarge the potential resources of drought-tolerant peanuts, we conducted in vitro mutagenesis with Pingyangmycin (PYM as the mutagen as well as directed screening on a medium supplemented with Hydroxyproline (HYP. After being extracted from mature seeds (cv. Huayu 20, the embryonic leaflets were cultured on somatic embryogenesis-induction medium with 4 mg/L PYM and the generated embryos were successively transferred to a germination medium with 4 and then 8 mmol/L HYP to screen HYP-tolerant plantlets. After that, these plantlets were grafted and transplanted to the experimental field. In the next generation, all seeds were sown in the field, and phenotype variation and trait segregation can be observed in most of the offspring (M2 generation. The M3 generation individuals were subjected to drought stress at the seedling stages. The activities of SOD and POD were substantially increased in eight offspring of 11 HYP-tolerant, regenerated plants than in their mutagenic parents. To determine the correlation between mutant phenotypes and genomic modification, we carried out a comparison of the DNA polymorphisms between the mutagenic parents and 13 M3 generation individuals from different HYP-tolerant, regenerated plants with SSR primers. Results showed that most mutants and parent plants had signs of polymorphisms. Under drought stress, some M3 generation individuals of 10 original HYP-tolerant, regenerated plants produced more pods than the mutagenic parent; twenty individuals among them produced >60 g pods/plant. M4-generation seeds were tested for quality characteristics by Near Infrared Spectroscopy (NIS and nine individuals with higher protein content (>30% and 21 individuals with higher oil content (>58% were screened. We concluded that the use of PYM-based in vitro mutagenesis in combination with directed screening with HYP is effective for the creation of potential drought-tolerant mutants of peanut.

  19. PHYTOCHEMICAL SCREENING AND IN VITRO ANTIOXIDANT ACTIVITY OF AQUEOUS AND HYDROALCOHOLIC EXTRACT OF BACOPA MONNIERI LINN.

    OpenAIRE

    Monic Shah et al

    2012-01-01

    The present study was undertaken to investigate in-vitro antioxidant activity of aqueous and hydroalcoholic extract of whole plant of Bacopa monnieri Linn. Family- Scrophularaceae. The total Phenolic content was determined using folin ciocalteau method while the total flavonoid content was determined using aluminium chloride method. In vitro antioxidant activity was evaluated using the Reducing power assay, Hydrogen peroxide scavenging assay, nitric oxide scavenging activity, super...

  20. Screening for Developmental Neurotoxicants using In Vitro "Brain on a Chip" Cultures

    Science.gov (United States)

    Currently there are thousands of chemicals in the environment that have not been screened for their potential to cause developmental neurotoxicity (DNT). The use of microelectrode array (MEA) technology allows for simultaneous extracellular measurement of action potential (spike)...

  1. Accurate and noninvasive embryos screening during in vitro fertilization (IVF) assisted by Raman analysis of embryos culture medium Accurate and noninvasive embryos screening during IVF

    Science.gov (United States)

    Shen, A. G.; Peng, J.; Zhao, Q. H.; Su, L.; Wang, X. H.; Hu, J. M.; Yang, J.

    2012-04-01

    In combination with morphological evaluation tests, we employ Raman spectroscopy to select higher potential reproductive embryos during in vitro fertilization (IVF) based on chemical composition of embryos culture medium. In this study, 57 Raman spectra are acquired from both higher and lower quality embryos culture medium (ECM) from 10 patients which have been preliminarily confirmed by clinical assay. Data are fit by using a linear combination model of least squares method in which 12 basis spectra represent the chemical features of ECM. The final fitting coefficients provide insight into the chemical compositions of culture medium samples and are subsequently used as criterion to evaluate the quality of embryos. The relative fitting coefficients ratios of sodium pyruvate/albumin and phenylalanine/albumin seem act as key roles in the embryo screening, attaining 85.7% accuracy in comparison with clinical pregnancy. The good results demonstrate that Raman spectroscopy therefore is an important candidate for an accurate and noninvasive screening of higher quality embryos, which potentially decrease the time-consuming clinical trials during IVF.

  2. The Identification of Genes Important in Pseudomonas syringae pv. phaseolicola Plant Colonisation Using In Vitro Screening of Transposon Libraries.

    Directory of Open Access Journals (Sweden)

    Bharani Manoharan

    Full Text Available The bacterial plant pathogen Pseudomonas syringae pv. phaseolicola (Pph colonises the surface of common bean plants before moving into the interior of plant tissue, via wounds and stomata. In the intercellular spaces the pathogen proliferates in the apoplastic fluid and forms microcolonies (biofilms around plant cells. If the pathogen can suppress the plant's natural resistance response, it will cause halo blight disease. The process of resistance suppression is fairly well understood, but the mechanisms used by the pathogen in colonisation are less clear. We hypothesised that we could apply in vitro genetic screens to look for changes in motility, colony formation, and adhesion, which are proxies for infection, microcolony formation and cell adhesion. We made transposon (Tn mutant libraries of Pph strains 1448A and 1302A and found 106/1920 mutants exhibited alterations in colony morphology, motility and biofilm formation. Identification of the insertion point of the Tn identified within the genome highlighted, as expected, a number of altered motility mutants bearing mutations in genes encoding various parts of the flagellum. Genes involved in nutrient biosynthesis, membrane associated proteins, and a number of conserved hypothetical protein (CHP genes were also identified. A mutation of one CHP gene caused a positive increase in in planta bacterial growth. This rapid and inexpensive screening method allows the discovery of genes important for in vitro traits that can be correlated to roles in the plant interaction.

  3. Sugar analog synthesis by in vitro biocatalytic cascade: A comparison of alternative enzyme complements for dihydroxyacetone phosphate production as a precursor to rare chiral sugar synthesis.

    Science.gov (United States)

    Hartley, Carol J; French, Nigel G; Scoble, Judith A; Williams, Charlotte C; Churches, Quentin I; Frazer, Andrew R; Taylor, Matthew C; Coia, Greg; Simpson, Gregory; Turner, Nicholas J; Scott, Colin

    2017-01-01

    Carbon-carbon bond formation is one of the most challenging reactions in synthetic organic chemistry, and aldol reactions catalysed by dihydroxyacetone phosphate-dependent aldolases provide a powerful biocatalytic tool for combining C-C bond formation with the generation of two new stereo-centres, with access to all four possible stereoisomers of a compound. Dihydroxyacetone phosphate (DHAP) is unstable so the provision of DHAP for DHAP-dependent aldolases in biocatalytic processes remains complicated. Our research has investigated the efficiency of several different enzymatic cascades for the conversion of glycerol to DHAP, including characterising new candidate enzymes for some of the reaction steps. The most efficient cascade for DHAP production, comprising a one-pot four-enzyme reaction with glycerol kinase, acetate kinase, glycerophosphate oxidase and catalase, was coupled with a DHAP-dependent fructose-1,6-biphosphate aldolase enzyme to demonstrate the production of several rare chiral sugars. The limitation of batch biocatalysis for these reactions and the potential for improvement using kinetic modelling and flow biocatalysis systems is discussed.

  4. In Silico Screening and In Vitro Activity Measurement of Javamide Analogues as Potential p38 MAPK Inhibitors

    Directory of Open Access Journals (Sweden)

    Jae B. Park

    2017-12-01

    Full Text Available p38 Mitogen-activated protein kinase (p38 MAPK is a protein kinase critically involved in the progress of inflammation/stress-associated diseases. Our data suggested that javamide analogues may contain strong anti-inflammation activities, but there is little information about their effects on p38 MAPK. Therefore, in this paper, the effects of thirty javamide analogues on p38 MAPK were investigated using in silico screening and in vitro p38 MAPK assay methods. The javamide analogues were synthesized and their chemical structures were confirmed using nuclear magnetic resonance (NMR spectroscopic methods. Then, the javamide analogues were screened using an in silico modeling program. The screened analogues demonstrated a wide range of binding energy (ΔE; −20 to −39 and several analogues with ΔE; −34 to −39 showed strong binding affinity to p38 MAPK. In vitro p38 MAPK assay, the kinase was significantly inhibited by the analogues with great binding energy (ΔE; −34 to −39 and in silico scores (Avg. score; −27.5 to −29.3. Furthermore, the comparative analysis of both assays showed a positive correlation between the in silico scores and p38 MAPK inhibition. In fact, the javamide analogues with top five in silico scores (Avg. score; −27.5 to −29.3 were found to inhibit p38 MAPK by 27–31% (p < 0.05 better than those with less scores (ΔE < −27.0. Especially, javamide-II-O-ethyl ester with relatively high in silico score (Avg. score; −29.2 inhibited p38 MAPK (IC50 = 9.9 μM a little better than its methyl ester with best in silico score (Avg. score; −29.3. To support the ability to inhibit p38 MAPK, the treatment of javamide-II-ethyl and -methyl esters could suppress the production of IL-8 and MCP-1 protein significantly by 22–73% (p < 0.05 in the differentiated THP-1 cells, and the inhibition was slightly stronger by the ethyl ester than the methyl ester. Altogether, this study suggests that javamide-II-O-ethyl ester may

  5. Formulation, high throughput in vitro screening and in vivo functional characterization of nanoemulsion-based intranasal vaccine adjuvants.

    Directory of Open Access Journals (Sweden)

    Pamela T Wong

    Full Text Available Vaccine adjuvants have been reported to induce both mucosal and systemic immunity when applied to mucosal surfaces and this dual response appears important for protection against certain pathogens. Despite the potential advantages, however, no mucosal adjuvants are currently approved for human use. Evaluating compounds as mucosal adjuvants is a slow and costly process due to the need for lengthy animal immunogenicity studies. We have constructed a library of 112 intranasal adjuvant candidate formulations consisting of oil-in-water nanoemulsions that contain various cationic and nonionic surfactants. To facilitate adjuvant development we first evaluated this library in a series of high-throughput, in vitro assays for activities associated with innate and adaptive immune activation in vivo. These in vitro assays screened for the ability of the adjuvant to bind to mucin, induce cytotoxicity, facilitate antigen uptake in epithelial and dendritic cells, and activate cellular pathways. We then sought to determine how these parameters related to adjuvant activity in vivo. While the in vitro assays alone were not enough to predict the in vivo adjuvant activity completely, several interesting relationships were found with immune responses in mice. Furthermore, by varying the physicochemical properties of the surfactant components (charge, surfactant polar head size and hydrophobicity and the surfactant blend ratio of the formulations, the strength and type of the immune response generated (TH1, TH2, TH17 could be modulated. These findings suggest the possibility of using high-throughput screens to aid in the design of custom adjuvants with unique immunological profiles to match specific mucosal vaccine applications.

  6. Identification of PPARgamma partial agonists of natural origin (I): development of a virtual screening procedure and in vitro validation.

    Science.gov (United States)

    Guasch, Laura; Sala, Esther; Castell-Auví, Anna; Cedó, Lidia; Liedl, Klaus R; Wolber, Gerhard; Muehlbacher, Markus; Mulero, Miquel; Pinent, Montserrat; Ardévol, Anna; Valls, Cristina; Pujadas, Gerard; Garcia-Vallvé, Santiago

    2012-01-01

    Although there are successful examples of the discovery of new PPARγ agonists, it has recently been of great interest to identify new PPARγ partial agonists that do not present the adverse side effects caused by PPARγ full agonists. Consequently, the goal of this work was to design, apply and validate a virtual screening workflow to identify novel PPARγ partial agonists among natural products. We have developed a virtual screening procedure based on structure-based pharmacophore construction, protein-ligand docking and electrostatic/shape similarity to discover novel scaffolds of PPARγ partial agonists. From an initial set of 89,165 natural products and natural product derivatives, 135 compounds were identified as potential PPARγ partial agonists with good ADME properties. Ten compounds that represent ten new chemical scaffolds for PPARγ partial agonists were selected for in vitro biological testing, but two of them were not assayed due to solubility problems. Five out of the remaining eight compounds were confirmed as PPARγ partial agonists: they bind to PPARγ, do not or only moderately stimulate the transactivation activity of PPARγ, do not induce adipogenesis of preadipocyte cells and stimulate the insulin-induced glucose uptake of adipocytes. We have demonstrated that our virtual screening protocol was successful in identifying novel scaffolds for PPARγ partial agonists.

  7. Identification of PPARgamma partial agonists of natural origin (I: development of a virtual screening procedure and in vitro validation.

    Directory of Open Access Journals (Sweden)

    Laura Guasch

    Full Text Available BACKGROUND: Although there are successful examples of the discovery of new PPARγ agonists, it has recently been of great interest to identify new PPARγ partial agonists that do not present the adverse side effects caused by PPARγ full agonists. Consequently, the goal of this work was to design, apply and validate a virtual screening workflow to identify novel PPARγ partial agonists among natural products. METHODOLOGY/PRINCIPAL FINDINGS: We have developed a virtual screening procedure based on structure-based pharmacophore construction, protein-ligand docking and electrostatic/shape similarity to discover novel scaffolds of PPARγ partial agonists. From an initial set of 89,165 natural products and natural product derivatives, 135 compounds were identified as potential PPARγ partial agonists with good ADME properties. Ten compounds that represent ten new chemical scaffolds for PPARγ partial agonists were selected for in vitro biological testing, but two of them were not assayed due to solubility problems. Five out of the remaining eight compounds were confirmed as PPARγ partial agonists: they bind to PPARγ, do not or only moderately stimulate the transactivation activity of PPARγ, do not induce adipogenesis of preadipocyte cells and stimulate the insulin-induced glucose uptake of adipocytes. CONCLUSIONS/SIGNIFICANCE: We have demonstrated that our virtual screening protocol was successful in identifying novel scaffolds for PPARγ partial agonists.

  8. Comparison of in vitro and in vivo screening models for androgenic and estrogenic activities

    NARCIS (Netherlands)

    Sonneveld, E.; Jansen, H.J.; Man, S.; Jonas, A.J.G.; Brouwer, A.; van der Burg, B.

    2006-01-01

    Identification of nuclear receptor-mediated endocrine activities is important in a variety of fields, ranging from pharmacological and clinical screening, to food and feed safety, toxicological monitoring, and risk assessment. Traditionally animal studies such as the Hershberger and Allen-Doisy

  9. High content screening as high quality assay for biological evaluation of photosensitizers in vitro.

    Directory of Open Access Journals (Sweden)

    Gisela M F Vaz

    Full Text Available A novel single step assay approach to screen a library of photdynamic therapy (PDT compounds was developed. Utilizing high content analysis (HCA technologies several robust cellular parameters were identified, which can be used to determine the phototoxic effects of porphyrin compounds which have been developed as potential anticancer agents directed against esophageal carcinoma. To demonstrate the proof of principle of this approach a small detailed study on five porphyrin based compounds was performed utilizing two relevant esophageal cancer cell lines (OE21 and SKGT-4. The measurable outputs from these early studies were then evaluated by performing a pilot screen using a set of 22 compounds. These data were evaluated and validated by performing comparative studies using a traditional colorimetric assay (MTT. The studies demonstrated that the HCS assay offers significant advantages over and above the currently used methods (directly related to the intracellular presence of the compounds by analysis of their integrated intensity and area within the cells. A high correlation was found between the high content screening (HCS and MTT data. However, the HCS approach provides additional information that allows a better understanding of the behavior of these compounds when interacting at the cellular level. This is the first step towards an automated high-throughput screening of photosensitizer drug candidates and the beginnings of an integrated and comprehensive quantitative structure action relationship (QSAR study for photosensitizer libraries.

  10. Agar composition affects in vitro screening of biocontrol activity of antagonistic microorganisms

    NARCIS (Netherlands)

    Bosmans, Lien; De Bruijn, I.; de Mot, Rene; Readers, Hans; Lievens, Bart

    2016-01-01

    Agar-based screening assays are the method of choice when evaluating antagonistic potential of bacterial biocontrol-candidates against pathogens.Weshowed thatwhen using the samemedium, but different agar compositions, the activity of a bacterial antagonist against Agrobacteriumwas strongly affected.

  11. Key learnings from performance of the U.S. EPA Endocrine Disruptor Screening Program (EDSP) Tier 1 in vitro assays.

    Science.gov (United States)

    LeBaron, Matthew J; Coady, Katie K; O'Connor, John C; Nabb, Diane L; Markell, Lauren K; Snajdr, Suzanne; Sue Marty, M

    2014-02-01

    Tier 1 of the U.S. EPA Endocrine Disruptor Screening Program comprises 11 studies: five in vitro assays, four in vivo mammalian assays, and two in vivo nonmammalian assays. The battery is designed to detect compounds with the potential to interact with the estrogen, androgen, or thyroid signaling pathways. This article examines the procedures, results, and data interpretation for the five Tier 1 in vitro assays: estrogen receptor (ER) and androgen receptor binding assays, an ER transactivation assay, an aromatase assay, and a steroidogenesis assay. Data are presented from two laboratories that have evaluated approximately 11 compounds in the Tier 1 in vitro assays. Generally, the ER and androgen receptor binding assays and the aromatase assay showed good specificity and reproducibility. As described in the guideline for the ER transactivation assay, a result is considered positive when the test compound induces a reporter gene signal that reaches 10% of the response seen with 1 nM 17β-estradiol (positive control). In the experience of these laboratories, this cutoff criterion may result in false-positive responses. For the steroidogenesis assay, there is variability in the basal and stimulated production of testosterone and estradiol by the H295R cells. This variability in responsiveness, coupled with potential cell stress at high concentrations of test compound, may make it difficult to discern whether hormone alterations are specific steroidogenesis alterations (i.e., endocrine active). Lastly, both laboratories had difficulty meeting some recommended performance criteria for each Tier 1 in vitro assay. Data with only minor deviations were deemed valid. © 2014 Wiley Periodicals, Inc.

  12. Development of assay platforms for in vitro screening of Treg modulating potential of pharmacological compounds

    DEFF Research Database (Denmark)

    Pedersen, Anders Elm; Holmstrøm, Kim; Jørgensen, Flemming

    2015-01-01

    sorting (FACS) sorted CD4 + CD25(high)CD127(dim/-)CD45RA+ naïve Treg cells followed by in vitro expansion. We report on the use of these cells in a short-term assay based on Treg mediated inhibition of the early effector T cell activation markers CD69 and CD154. Additionally, we investigate the use...

  13. In vitro microtumors provide a physiologically predictive tool for breast cancer therapeutic screening.

    Directory of Open Access Journals (Sweden)

    Gabriel Benton

    Full Text Available Many anti-cancer drugs fail in human trials despite showing efficacy in preclinical models. It is clear that the in vitro assays involving 2D monoculture do not reflect the complex extracellular matrix, chemical, and cellular microenvironment of the tumor tissue, and this may explain the failure of 2D models to predict clinical efficacy. We first optimized an in vitro microtumor model using a tumor-aligned ECM, a tumor-aligned medium, MCF-7 and MDA-MB-231 breast cancer spheroids, human umbilical vein endothelial cells, and human stromal cells to recapitulate the tissue architecture, chemical environment, and cellular organization of a growing and invading tumor. We assayed the microtumor for cell proliferation and invasion in a tumor-aligned extracellular matrix, exhibiting collagen deposition, acidity, glucose deprivation, and hypoxia. We found maximal proliferation and invasion when the multicellular spheroids were cultured in a tumor-aligned medium, having low pH and low glucose, with 10% fetal bovine serum under hypoxic conditions. In a 7-day assay, varying doses of fluorouracil or paclitaxel had differential effects on proliferation for MCF-7 and MDA-MB-231 tumor spheroids in microtumor compared to 2D and 3D monoculture. The microtumors exhibited a tumor morphology and drug response similar to published xenograft data, thus demonstrating a more physiologically predictive in vitro model.

  14. Acanthamoeba castellanii: a new high-throughput method for drug screening in vitro

    OpenAIRE

    Ortega-Rivas, Antonio; Padrón, José M; Valladares, Basilio; Elsheikha, Hany M

    2016-01-01

    Despite significant public health impact, there is no specific antiprotozoal therapy for prevention and treatment of Acanthamoeba castellanii infection. There is a need for new and efficient anti-Acanthamoeba drugs that are less toxic and can reduce treatment duration and frequency of administration. In this context a new, rapid and sensitive assay is required for high-throughput activity testing and screening of new therapeutic compounds. A colorimetric assay based on sulforhodamine B (SRB) ...

  15. In vitro screens for quorum sensing inhibitors and in vivo confirmation of their effect

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; van Gennip, Maria; Jakobsen, Tim H

    2010-01-01

    , with the aim of disabling the expression of virulence and reduction of antibiotic tolerance. Here we present protocols for screening and testing for acyl-homoserine lactone (AHL)-dependent QS inhibition. We also present protocols for the in vivo validation of QSIs as possible drug candidates. The presented...... methods allow the evaluation of QS inhibition by a potential drug candidate within 2-3 weeks....

  16. Assessment of beating parameters in human induced pluripotent stem cells enables quantitative in vitro screening for cardiotoxicity

    Energy Technology Data Exchange (ETDEWEB)

    Sirenko, Oksana, E-mail: oksana.sirenko@moldev.com [Molecular Devices LLC, Sunnyvale, CA 94089 (United States); Cromwell, Evan F., E-mail: evan.cromwell@moldev.com [Molecular Devices LLC, Sunnyvale, CA 94089 (United States); Crittenden, Carole [Molecular Devices LLC, Sunnyvale, CA 94089 (United States); Wignall, Jessica A. [Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill, NC 27599 (United States); Wright, Fred A. [Department of Biostatistics, University of North Carolina, Chapel Hill, NC 27599 (United States); Rusyn, Ivan [Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill, NC 27599 (United States)

    2013-12-15

    Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes show promise for screening during early drug development. Here, we tested a hypothesis that in vitro assessment of multiple cardiomyocyte physiological parameters enables predictive and mechanistically-interpretable evaluation of cardiotoxicity in a high-throughput format. Human iPSC-derived cardiomyocytes were exposed for 30 min or 24 h to 131 drugs, positive (107) and negative (24) for in vivo cardiotoxicity, in up to 6 concentrations (3 nM to 30 uM) in 384-well plates. Fast kinetic imaging was used to monitor changes in cardiomyocyte function using intracellular Ca{sup 2+} flux readouts synchronous with beating, and cell viability. A number of physiological parameters of cardiomyocyte beating, such as beat rate, peak shape (amplitude, width, raise, decay, etc.) and regularity were collected using automated data analysis. Concentration–response profiles were evaluated using logistic modeling to derive a benchmark concentration (BMC) point-of-departure value, based on one standard deviation departure from the estimated baseline in vehicle (0.3% dimethyl sulfoxide)-treated cells. BMC values were used for cardiotoxicity classification and ranking of compounds. Beat rate and several peak shape parameters were found to be good predictors, while cell viability had poor classification accuracy. In addition, we applied the Toxicological Prioritization Index (ToxPi) approach to integrate and display data across many collected parameters, to derive “cardiosafety” ranking of tested compounds. Multi-parameter screening of beating profiles allows for cardiotoxicity risk assessment and identification of specific patterns defining mechanism-specific effects. These data and analysis methods may be used widely for compound screening and early safety evaluation in drug development. - Highlights: • Induced pluripotent stem cell-derived cardiomyocytes are promising in vitro models. • We tested if evaluation

  17. A mix-and-read drop-based in vitro two-hybrid method for screening high-affinity peptide binders

    OpenAIRE

    Naiwen Cui; Huidan Zhang; Nils Schneider; Ye Tao; Haruichi Asahara; Zhiyi Sun; Yamei Cai; Koehler, Stephan A.; de Greef, Tom F. A.; Alireza Abbaspourrad; Weitz, David A; Shaorong Chong

    2016-01-01

    Drop-based microfluidics have recently become a novel tool by providing a stable linkage between phenotype and genotype for high throughput screening. However, use of drop-based microfluidics for screening high-affinity peptide binders has not been demonstrated due to the lack of a sensitive functional assay that can detect single DNA molecules in drops. To address this sensitivity issue, we introduced in vitro two-hybrid system (IVT2H) into microfluidic drops and developed a streamlined mix-...

  18. 3D Printing of Tissue Engineered Constructs for In Vitro Modeling of Disease Progression and Drug Screening.

    Science.gov (United States)

    Vanderburgh, Joseph; Sterling, Julie A; Guelcher, Scott A

    2017-01-01

    2D cell culture and preclinical animal models have traditionally been implemented for investigating the underlying cellular mechanisms of human disease progression. However, the increasing significance of 3D vs. 2D cell culture has initiated a new era in cell culture research in which 3D in vitro models are emerging as a bridge between traditional 2D cell culture and in vivo animal models. Additive manufacturing (AM, also known as 3D printing), defined as the layer-by-layer fabrication of parts directed by digital information from a 3D computer-aided design file, offers the advantages of simultaneous rapid prototyping and biofunctionalization as well as the precise placement of cells and extracellular matrix with high resolution. In this review, we highlight recent advances in 3D printing of tissue engineered constructs that recapitulate the physical and cellular properties of the tissue microenvironment for investigating mechanisms of disease progression and for screening drugs.

  19. Efficient and fast functional screening of microdystrophin constructs in vivo and in vitro for therapy of duchenne muscular dystrophy

    DEFF Research Database (Denmark)

    Jørgensen, Louise Helskov; Larochelle, Nancy; Orlopp, Kristian

    2009-01-01

    Duchenne muscular dystrophy (DMD) is an X-linked, lethal genetic disorder affecting the skeletal muscle compartment, and is caused by mutation(s) in the dystrophin gene. Gene delivery of microdystrophin constructs using adeno-associated virus (AAV) and antisense-mediated exon skipping restoring...... the genetic reading frame are two of the most promising therapeutic strategies for DMD. Both approaches use microdystrophin proteins either directly as a desired construct for gene delivery, using the capacity-limited AAV vectors, or as the therapeutic outcome of gene splicing. Although functionality...... of the resulting artificial dystrophin proteins can be predicted in silico, experimental evidence usually obtained in transgenic mice is required before human trials. However, the enormous number of potential constructs makes screening assays for dystrophin protein function in vitro and in vivo highly desirable...

  20. Phytochemical screening of Nepeta cataria extracts and their in vitro inhibitory effects on free radicals and carbohydrate-metabolising enzymes.

    Science.gov (United States)

    Naguib, Abdel Moneam Mohamed; Ebrahim, Mohamed Elsayed; Aly, Hanan Farouk; Metawaa, Hemaia Mohamed; Mahmoud, Ahlam Hosni; Mahmoud, Ebtissam A; Ebrahim, Faten Mohamed

    2012-01-01

    This research was performed to investigate in vitro the biological activities of successive as well as 70% ethanol extracts of Nepeta cataria on some biochemical parameters including oxidative markers and carbohydrate-hydrolysing enzyme activities (α-amylase, β-galactosidase and α-glucosidase). Powdered N. cataria and its successive extracts were screened for their phytochemical constituents. Tests for tannins, carbohydrates, glycosides and flavonoids were positive in ethanolic extract, but those for steroids and terpenoids were positive in petroleum ether and chloroform extracts. Also, different extracts were chromatographically investigated. The results obtained demonstrated that different successive extracts of N. cataria exhibited an inhibitory effect on oxidative stress indices and carbohydrate-hydrolysing enzymes. It is observed that 70% ethanol, petroleum ether and chloroform extracts showed, respectively, the most potent inhibitory activities, while ethyl acetate and ethanol successive extracts appeared with moderate or low reducing activities.

  1. Novel inhibitors of dengue virus methyltransferase: discovery by in vitro-driven virtual screening on a desktop computer grid.

    Science.gov (United States)

    Podvinec, Michael; Lim, Siew Pheng; Schmidt, Tobias; Scarsi, Marco; Wen, Daying; Sonntag, Louis-Sebastian; Sanschagrin, Paul; Shenkin, Peter S; Schwede, Torsten

    2010-02-25

    Dengue fever is a viral disease that affects 50-100 million people annually and is one of the most important emerging infectious diseases in many areas of the world. Currently, neither specific drugs nor vaccines are available. Here, we report on the discovery of new inhibitors of the viral NS5 RNA methyltransferase, a promising flavivirus drug target. We have used a multistage molecular docking approach to screen a library of more than 5 million commercially available compounds against the two binding sites of this enzyme. In 263 compounds chosen for experimental verification, we found 10 inhibitors with IC(50) values of <100 microM, of which four exhibited IC(50) values of <10 microM in in vitro assays. The initial hit list also contained 25 nonspecific aggregators. We discuss why this likely occurred for this particular target. We also describe our attempts to use aggregation prediction to further guide the study, following this finding.

  2. In vitro phenotypic screening of 7-chloro-4-amino(oxy)quinoline derivatives as putative anti-Trypanosoma cruzi agents.

    Science.gov (United States)

    Fonseca-Berzal, Cristina; Rojas Ruiz, Fernando A; Escario, José A; Kouznetsov, Vladimir V; Gómez-Barrio, Alicia

    2014-02-15

    In this study, a series of 22 pre-synthesized 7-chloro-4-amino(oxy)quinoline derivatives was assayed in vitro as potential antichagasic agents. A primary screening against Trypanosoma cruzi epimastigotes and a non-specific cytotoxicity assay on murine fibroblasts were simultaneously performed, resulting quinolines 3, 7 and 12 with great selectivity (SI) on the extracellular parasite (SI7, SI3, SI12 and SIBZ >9.44). Therefore, the activity of these derivatives was evaluated on intracellular amastigotes, achieving derivative 7 the best SI (SI=12.73). These results, supported by the in silico prediction of a good oral bioavailability and a suitable risk profile, propose the 4-amino-7-chloroquinoline scaffold as a potential template for designing trypanocidal prototypes. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. 3D Printing of Tissue Engineered Constructs for in vitro Modeling of Disease Progression and Drug Screening

    Science.gov (United States)

    Vanderburgh, Joseph; Sterling, Julie A.

    2016-01-01

    2D cell culture and preclinical animal models have traditionally been implemented for investigating the underlying cellular mechanisms of human disease progression. However, the increasing significance of 3D versus 2D cell culture has initiated a new era in cell culture research in which 3D in vitro models are emerging as a bridge between traditional 2D cell culture and in vivo animal models. Additive manufacturing (AM, also known as 3D printing), defined as the layer-by-layer fabrication of parts directed by digital information from a 3D computer-aided design (CAD) file, offers the advantages of simultaneous rapid prototyping and biofunctionalization as well as the precise placement of cells and extracellular matrix with high resolution. In this review, we highlight recent advances in 3D printing of tissue engineered constructs (TECs) that recapitulate the physical and cellular properties of the tissue microenvironment for investigating mechanisms of disease progression and for screening drugs. PMID:27169894

  4. IN VITRO MASS-SCREENING OF LACTIC ACID BACTERIA AS POTENTIAL BIOSORBENTS OF CESIUM AND STRONTIUM IONS

    Directory of Open Access Journals (Sweden)

    Hideki Kinoshita

    2015-04-01

    Full Text Available Many radionuclides were scattered by the explosion at the Fukushima Daiichi Nuclear Power Station. We examined whether lactic acid bacteria (LAB can sorb cesium ions (Cs+ and strontium ions (Sr2+ for radioprotection. Many strains showed biosorption to Cs+ and Sr2+ using an in vitro mass-screening although each strain showed different sorption. We selected MYU 111, MYU 758, and MYU 759 strains that showed especially high biosorption to Cs+ and/or Sr2+. MYU 111 was identified as Lactobacillus plantarum, and MYU 758 and 759 were Pediococcus pentosaceus. The selected strains tended to show higher biosorption when using the buffer method compared to the culture method. Further, they showed high biosorption at a low concentration of 1 ppb Cs+ and Sr2+ (max 28.8% and 97.7% sorption, respectively. This is the first study where lactic acid bacteria are shown to have biosorption of Cs+ and Sr2+.

  5. Virtual screening, identification and in vitro testing of novel inhibitors of O-acetyl-L-serine sulfhydrylase of Entamoeba histolytica.

    Directory of Open Access Journals (Sweden)

    Isha Nagpal

    Full Text Available The explosive epidemicity of amoebiasis caused by the facultative gastrointestinal protozoan parasite Entamoeba histolytica is a major public health problem in developing countries. Multidrug resistance and side effects of various available antiamoebic drugs necessitate the design of novel antiamobeic agents. The cysteine biosynthetic pathway is the critical target for drug design due to its significance in the growth, survival and other cellular activities of E. histolytica. Here, we have screened 0.15 million natural compounds from the ZINC database against the active site of the EhOASS enzyme (PDB ID. 3BM5, 2PQM, whose structure we previously determined to 2.4 Å and 1.86 Å resolution. For this purpose, the incremental construction algorithm of GLIDE and the genetic algorithm of GOLD were used. We analyzed docking results for top ranking compounds using a consensus scoring function of X-Score to calculate the binding affinity and using ligplot to measure protein-ligand interactions. Fifteen compounds that possess good inhibitory activity against EhOASS active site were identified that may act as potential high affinity inhibitors. In vitro screening of a few commercially available compounds established their biological activity. The first ranked compound ZINC08931589 had a binding affinity of ∼8.05 µM and inhibited about 73% activity at 0.1 mM concentration, indicating good correlation between in silico prediction and in vitro inhibition studies. This compound is thus a good starting point for further development of strong inhibitors.

  6. Acanthamoeba castellanii: A new high-throughput method for drug screening in vitro.

    Science.gov (United States)

    Ortega-Rivas, Antonio; Padrón, José M; Valladares, Basilio; Elsheikha, Hany M

    2016-12-01

    Despite significant public health impact, there is no specific antiprotozoal therapy for prevention and treatment of Acanthamoeba castellanii infection. There is a need for new and efficient anti-Acanthamoeba drugs that are less toxic and can reduce treatment duration and frequency of administration. In this context a new, rapid and sensitive assay is required for high-throughput activity testing and screening of new therapeutic compounds. A colorimetric assay based on sulforhodamine B (SRB) staining has been developed for anti-Acanthamoeba drug susceptibility testing and adapted to a 96-well microtiter plate format. Under these conditions chlorhexidine was tested to validate the assay using two clinical strains of A. castellanii (Neff strain, T4 genotype [IC50 4.68±0.6μM] and T3 genotype [IC50 5.69±0.9μM]). These results were in good agreement with those obtained by the conventional Alamar Blue assay, OCR cytotoxicity assay and manual cell counting method. Our new assay offers an inexpensive and reliable method, which complements current assays by enhancing high-throughput anti-Acanthamoeba drug screening capabilities. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. In vitro Antimicrobial Activity ofCitrus aurantifolia and its Phytochemical screening

    Directory of Open Access Journals (Sweden)

    Rafi khan Pathan

    2012-05-01

    Full Text Available Objective: To evaluate the antimicrobial efficacy of Citrus aurantifolia Linn (CA against some microorganisms - bacteria and fungus were Staphylococcus aureus, Escherichia coli, Klebsiella pneumonia, Pseudomonas spp, Aspergillus niger, Aspergillus fumigates, Mucor spp and Pencillium. Methods: 100 毺 l of 10 mg CA were assessed against eight test microorganisms by agar well Diffusion Method. Gentamicin and Ketoconazole 10 mg/ml were used as standards. A different solvent was used to obtain CA leaf extract by using maceration technique. Results: %yield obtained for dried leaf extract of CA with chloroform, ethanol, acetone, petroleum ether and aqueous ethanol was approximately 15%, 18%, 09%, 11% and 24% respectively. Due to its high yield value hydroalcoholic extract of CA was used for estimating the antimicrobial activity and its phytochemical screening. Phytochemical screening of CA plant reveals the presence of Alkaloids, carbohydrates, flavonoids, steroids and tannins. Conclusions: The study demonstrates that the hydroalcoholic extract of CA leaf exhibit antibacterial activity on Klebsiella pneumonia, Pseudomonas sp, Staphylococcus aureus and antifungal activity among Aspergillus niger, Aspergillus fumigates, Mucor species. These recognized a good support to the use of this plant in herbal medicine and as base for the development of new drugs and phytomedicine.

  8. Preliminary phytochemical screening and in vitro antibacterial activity of Anamirta cocculus (Linn. seeds

    Directory of Open Access Journals (Sweden)

    Umer Qadir

    2015-04-01

    Full Text Available The present work is aimed at primary screening of the phytochemical contents of seven solvents (petroleum ether, benzene, chloroform, acetone, methanol, ethanol and a mixture of methanol and ethyl acetate (1:3 extracts of the seeds of Anamirta cocculus (Linn. and to evaluate the antibacterial activity of these extracts against five species of pathogenic bacteria. The powdered seeds of A. cocculus were extracted with seven solvents (polar and non-polar with Soxhlet apparatus and the extracts were subjected to preliminary phytochemical screening by standard biochemical tests. The antibacterial activity of various extracts was evaluated against five bacterial species (Staphylococcus aureus, Proteus vulgaris, Escherichia coli, Salmonella typhi and Klebsiella pneumoniae. The zone of inhibitions and minimum inhibitory concentration (MIC of the various extracts were determined by running the experiments in triplicates. The extracts of the seeds of A. cocculus contained various pharmaceutically active substances viz., aldehydes, alkaloids, phenolic compounds, flavonoids, saponins, carbohydrates, proteins, lipids, glycosides, phytosterols, volatile oils, gums and mucilage and other minor phytochemicals. All the extracts showed significant levels of antibacterial activity. Methanol extract was the most active one with remarkable antibacterial activity on the various species tested. MICs of the extracts revealed methanol extract as the most potent one with the lowest inhibitory concentration of 3 μg/mL on E. coli. The findings of the present study indicated that the seeds of A. cocculus possess various secondary metabolites having the potential for developing pharmaceutical drugs, especially antimicrobial ones.

  9. High throughput screening (HTS) for phototoxicity hazard using the in vitro 3T3 neutral red uptake assay.

    Science.gov (United States)

    Jones, P A; King, A V

    2003-01-01

    Testing for phototoxic hazard is usually carried out for product ingredients intended for use on skin, which may be exposed to sunlight. Unilever currently uses the validated in vitro 3T3 Neutral Red Uptake phototoxicity test (NRU PT). This protocol involves 2-3 experiments, each taking 3 days to perform. One person can test up to seven test materials plus positive control at any one time, requiring approximately 0.5 g test material. Higher throughput is required where libraries of potential actives are being generated and screening for potential phototoxicants is required. A proposed HTS protocol would use the NRU PT, but only one concentration (10 microg/ml) in a single experiment. The validity of the HTS protocol was investigated by a retrospective examination of data from 86 materials previously tested. Phototoxic hazard predictions made using the conventional NRU PT were compared with those obtained if only data at 10 microg/ml were considered. A majority of 73 materials (84.9%) gave agreement in predictions between the two protocols; for 13 materials (15.1%) the assessments did not agree. There were no false positives; however, there were some false negatives, i.e., predicted as phototoxic from the conventional assay, but non-phototoxic at 10 microg/ml. As this protocol is intended for screening purposes only it is considered that this would be acceptable at this stage in material selection. One person could screen 128 test materials in 3 days, requiring selected for further development and inclusion in a formulation may require further confirmatory testing, e.g. using a human skin model assay for phototoxicity.

  10. Screening lactic acid bacteria from swine origins for multistrain probiotics based on in vitro functional properties.

    Science.gov (United States)

    Guo, Xiao-Hua; Kim, Jong-Man; Nam, Hyang-Mi; Park, Shin-Young; Kim, Jae-Myung

    2010-08-01

    Lactic acid bacteria originated from swine feces and intestines were selected for potential probiotics based on their bile-salt resistance, low pH tolerance, potential adhesion to epithelial cells and especially functional properties, including production of antimicrobial substances, bile-salt hydrolase (BSH) and amylolytic activity. Results showed 7 isolates with antimicrobial activity, 5 with BSH activity and 3 with amylolytic activity were preliminarily selected from 485 lactic acid bacteria based on their highest potential with functional properties in vitro. The 15 isolates were further assayed on the essential characteristics as potential probiotics. All isolates were fully tolerant to 0.3% bile salts and 11 of them were able to resist pH 3 for 3 h without loss of viable cells. The eleven isolates were then evaluated on their adhesion capability. Wide variation in the hydrophobic character and specific adhesion efficiency was observed and three isolates G1-1, G22-2 and G8-5, with respective antimicrobial, BSH and amylolytic activities were finally selected. In addition, the three isolates were compatible in the coexistence assay. Isolate G1-1 was identified as Lactobacillus salivarius by API system and a 16S rRNA gene sequence analysis. Both G8-5 and G22-2 showed the closest homology to Lactobacillus reuteri according to their 16S rRNA gene sequences (99%). From the study, the three Lactobacilli strains were shown to share the functional properties necessary for probiotics use in animal additives. Their compatibility with respective in vitro activities was expected to show enhanced in vivo efficacy after combination for multistrain probiotics use. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

  11. In Vitro Screening of 1877 Industrial and Consumer Chemicals, Pesticides and Pharmaceuticals in up to 782 Assays: ToxCast Phase I and II (SOT)

    Science.gov (United States)

    In Phase II of the ToxCast program, the U.S. EPA and Tox21 partners screened 1,877 chemicals, including pesticides; food, cosmetics and personal care ingredients; pharmaceuticals; and industrial chemicals. Testing used a 782 in vitro assays across 7 technologies and multiple bi...

  12. Multivariate toxicity profiles and QSAR modelling of NDL-PCBs – an investigation of in vitro screening data from ultra-pure congeners

    NARCIS (Netherlands)

    Stenberg, M.; Hamers, T.H.M.; Machala, M.; Fonnum, F.; Stenius, U.; Al Anati, L.; van Duursen, M.; Westerink, R.H.S.; Fernandes, E.C.A.; Andersson, P.L.

    2011-01-01

    The non-dioxin-like PCBs (NDL-PCBs) found in food and human samples have a complex spectrum of adverse effects, but lack a detailed risk assessment. The toxicity profiles of 21 carefully selected PCBs (19 NDL-PCBs) were identified by in vitro screening in 17 different assays on specific endpoints

  13. Probe molecules (PrM) approach in adverse outcome pathway (AOP) based high throughput screening (HTS): in vivo discovery for developing in vitro target methods

    Science.gov (United States)

    Efficient and accurate adverse outcome pathway (AOP) based high-throughput screening (HTS) methods use a systems biology based approach to computationally model in vitro cellular and molecular data for rapid chemical prioritization; however, not all HTS assays are grounded by rel...

  14. Stretch and/or oxygen glucose deprivation (OGD in an in vitro traumatic brain injury (TBI model induces calcium alteration and inflammatory cascade

    Directory of Open Access Journals (Sweden)

    Ellaine eSalvador

    2015-08-01

    Full Text Available The blood-brain barrier (BBB, made up of endothelial cells of capillaries in the brain, maintains the microenvironment of the central nervous system. During ischemia and traumatic brain injury (TBI, cellular disruption leading to mechanical insult results to the BBB being compromised. Oxygen glucose deprivation (OGD is the most commonly used in vitro model for ischemia. On the other hand, stretch injury is currently being used to model TBI in vitro. In this paper, the two methods are used alone or in combination, to assess their effects on cerebrovascular endothelial cells cEND in the presence or absence of astrocytic factors. Applying severe stretch and/or OGD to cEND cells in our experiments resulted to cell swelling and distortion. Damage to the cells induced release of lactate dehydrogenase enzyme (LDH and nitric oxide (NO into the cell culture medium. In addition, mRNA expression of inflammatory markers interleukin (IL-6, IL-1α, chemokine (C-C motif ligand 2 (CCL2 and tumor necrosis factor (TNF-α also increased. These events could lead to the opening of calcium ion channels resulting to excitotoxicity. This could be demonstrated by increased calcium level in OGD-subjected cEND cells incubated with astrocyte-conditioned medium. Furthermore, reduction of cell membrane integrity decreased tight junction proteins claudin-5 and occludin expression. In addition, permeability of the endothelial cell monolayer increased. Also, since cell damage requires an increased uptake of glucose, expression of glucose transporter glut1 was found to increase at the mRNA level after OGD. Overall, the effects of OGD on cEND cells appear to be more prominent than that of stretch with regards to TJ proteins, NO, glut1 expression and calcium level. Astrocytes potentiate these effects on calcium level in cEND cells. Combining both methods to model TBI in vitro shows a promising improvement to currently available models.

  15. In vitro screening of radioprotective properties in the novel glucosylated flavonoids.

    Science.gov (United States)

    Yu, Hao; Haskins, Jeremy S; Su, Cathy; Allum, Allison; Haskins, Alexis H; Salinas, Victoria A; Sunada, Shigeaki; Inoue, Taisuke; Aizawa, Yasushi; Uesaka, Mitsuru; Kato, Takamitsu A

    2016-11-01

    Novel glucosyl flavonoids are developed by the addition of glucose to naturally occurring flavonoids. Flavonoids are known antioxidants that possess radioprotective properties. In order to investigate the radioprotective properties of novel glucosyl flavonoids, in vitro DNA double-strand breaks (DSBs) analysis was carried out. In the present study, Quercetin, Naringenin, and Hesperetin groups of flavonoids included in the natural and novel glucosyl 13 flavonoids were investigated. Flavonoids were mixed with Lambda DNA, and subsequently exposed to gamma‑rays. Furthermore, DNA DSB yields were visualized by gel electrophoresis. Quercetin derivatives displayed reduced DNA DSB formation at 10 µM. At a high concentration, the majority of flavonoids displayed radioprotective properties as a reduction of DSB yields. Suppression of DSB formation was confirmed via the molecular combing assay for Quercetin, and three monoglucosyl flavonoids. Glucosylation showed positive effects for radioprotection and monoglucosyl-Rutin showed superior radioprotective properties when compared to monoglucosyl-Naringin and Hesperidin. In addition, Quercetin derivatives had greater total antioxidant capacities and DPPH radical scavenging ability than other flavonoid groups. Since Quercetin, Isoquercetin, and Rutin display poor water solubility, monoglucosyl-Rutin, maltooligosyl-Isoquercetin, and maltooligosyl-Rutin may be better radioprotective agents and easily bioavailable with increased water solubility.

  16. Chemical Composition and in Vitro Antifungal Activity Screening of the Allium ursinum L. (Liliaceae

    Directory of Open Access Journals (Sweden)

    Radu Vasile Bagiu

    2012-01-01

    Full Text Available The objective of the study was to summarize the methods for isolating and identifying natural sulfur compounds from Allium ursinum (ramson and to discuss the active constituents with regard to antifungal action. Using chromatographic techniques, the active constituents were isolated and subsequently identified. Analyses by high-performance liquid chromatography (HPLC suggested that these compounds were sulfur constituents, with a characteristic absorbance at 250 nm. Gas chromatography-mass spectrometry (GC-MS analyses allowed the chemical structures of the isolated constituents to be postulated. We adopted the same methods to identify the health-giving profiling of ramsons and the effects are thought to be primarily derived from the presence and breakdown of the alk(enylcysteine sulphoxide, alliin and its subsequent breakdown to allicin (sulfur-compounds of ramson in connection with antifungal action. The aim of the study was the characterization of the chemical composition of ramsons and the testing of the action of the in vitro extracts, on different strains of Candida albicans. The main goal was to highlight the most efficient extracts of Allium ursinum that can provide long-term antifungal activity without remissions. The extracts from Allium ursinum plants, inhibited growth of Candida spp. cells at concentrations ranging from 0.5 to 4.0 mg/mL, while that of adherent cells at concentrations ranging from 1.0 to > 4.0 mg/mL, depending on the yeast and plant species.

  17. In vitro Biphasic Effect of Honey Bee Venom on Basophils from Screened Healthy Blood Donors.

    Science.gov (United States)

    Chirumbolo, Salvatore; Zanoni, Giovanna; Ortolani, Riccardo; Vella, Antonio

    2011-01-01

    Apis mellifera L. bee venom is the most studied hymenoptera allergen, but many aspects of its action on human basophils remain unclear. Allergologists seek evidence of the effectiveness of bee venom immunotherapy as this approach is the chosen treatment for systemic allergic reactions. The effect of bee venom on human basophils in vitro has not been studied in detail for many reasons, including the paucity of basophils in peripheral blood, inter-individual basophil response variability, and the reliability and predictability of basophil activation tests. We conducted a brief preliminary survey of the effect of Apis bee venom on healthy asymptomatic (non-allergic) subjects. A dose of an aqueous commercial extract of Apis bee venom as high as 10 µg/mL activated resting basophils (CD63=+80-90%, CD203c=+30%), while it inhibited the expression of CD63 (-50%) following basophil stimulation by the soluble agonists formyl-Met-Leu-Phe or anti-IgE. The activation of resting basophils appeared to be dose-related. Only when basophils were activated with an IgE-mediated agonist, did bee venom extract exhibit a possible priming mechanism at the lowest doses used only via CD63, while it was ineffective via CD203c. Autocrine interleukin-3 may play a role in the observed biphasic behavior.

  18. Biological Screening of Eichornia crassipes against Different Pathogenic Microbes: An In Vitro Study

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    Rubina Rehman

    2016-09-01

    Full Text Available The present research is a biological screening of Eichornia crassipes (Pontederiaceae. Dichloromethane and methanol extracts of the whole plant were investigated for their antibacterial, antifungal, phytotoxic, and cytotoxic activities. The antibacterial activity was evaluated using agar well-diffusion method against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Shigella flexneri, Pseudomonas aeruginosa, and Salmonella typhi. The antifungal activity was evaluated using the agar tube–dilution method against Candida albicans, Candida glabrata, Aspergillus flavus, Microsporum canis, and Fusarium solani. The phytotoxicity activity was determined using Lemna bioassay against Lemna minor. Brine shrimp–cytotoxicity assay was determined against brine-shrimp larvae. Dichloromethane extract exhibited significant phytotoxicity (100% growth regulation at 1,000 µg/ml concentration against Lemna minor whereas methanolic extracts showed moderate (75% growth regulation phytotoxicity at the same concentration. Methanolic extract showed cytotoxicity at the highest level of dose whereas dichloromethane extract showed no activity having Etoposide as standard drug. Both of the extracts have nonsignificant antifungal and antibacterial activity.

  19. Preliminary phytochemical screening and in vitro antibacterial activity of Bauhinia variegata Linn. against human pathogens

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    Sonam Pandey

    2015-02-01

    Full Text Available Objective: To investigate the antimicrobial and phytochemical properties of hydromethanolic extracts of Bauhinia variegata Linn. (B. variegata (leaf, stem bark and flower to justify the traditional claim endowed upon this herbal drug as a rasayana in Ayurveda. This study thus can be further utilized to formulate the natural antioxidant which can be used as a dietary supplement to fight against several diseases such as cancer, ageing, arthrosclerosis, etc. Methods: The study showed that the number of different phytoconstituents present in the plant which makes it remarkable for its use by traditional practitioners. On the another set of experiment, the hydromethanolic extract of B. variegata (leaf, stem bark and flower were evaluated against Gram-positive and Gram-negative by using disk diffusion assay. Results: Phytochemical screening of all extracts showed the presence of alkaloids, steroids, phenolic compounds, tannins, saponin, carbohydrates, proteins, amino acids and organic acids. The antibacterial activity of all the extracts (leaf, stem bark and flower of B. variegata was determined by agar well diffusion method at four different concentrations i.e., 1 000 mg/mL, 750 mg/mL, 500 mg/mL and 250 mg/mL using Gram-positive Bacillus subtilius, Staphylococcus aureus and Streptococcus epidermidis and Gram-negative Escherichia coli, Shegilla flexineria, Pseudomonas auriginosa bacteria. Conclusions: These studies show that hydromethanolic extracts of B. variegata (leaf, stem bark and flower inhibited the growth of microorganism ’s in dose dependently. B. variegata leaf, stem bark and flower extracts have several phytochemical constituents who possess the antimicrobial activity. A tiny amount of data is presented, as the preliminary antimicrobial properties of the B. variegata here accessed, under the urgent necessity of new antibiotics in the market and in face of the increased resistance of infectious microorganisms to antimicrobials.

  20. Screening of Nepalese crude drugs traditionally used to treat hyperpigmentation: in vitro tyrosinase inhibition.

    Science.gov (United States)

    Adhikari, A; Devkota, H P; Takano, A; Masuda, K; Nakane, T; Basnet, P; Skalko-Basnet, N

    2008-10-01

    South-East Asian population is daily exposed to strong sunlight. As a result, the majority of population will have darker, ethnic skin. Moreover, many people suffer from dark spots, hyperpigmentation, which is considered to be a skin disorder and causes psychological disturbance. To treat dark spots, most of the population will still rely on traditionally used crude drugs, knowledge about which is transferred from generation to generation. Fifty-two crude drugs were selected based on the survey performed among local healers and beauticians of different ethnic origin. These crude drugs were screened for mushroom tyrosinase inhibitory activity, as tyrosinase inhibitors are becoming increasingly important as cosmetic and medicinal products, primarily to control hyperpigmentation. Among the tested crude drugs, methanolic extracts of Glycyrrhiza glabra, Morus alba, Syzygium aromaticum, Citrus aurantifolia, Cypreae moneta, Punica granatum and Citrus aurantium, at the final concentration of 50 microg mL(-1), showed mushroom tyrosinase inhibitory activity of 78.9%, 71.0%, 69.4%, 59.0%, 56.0%, 53.4 and 51.9%, respectively, with 91.4% inhibitory activity of kojic acid taken as positive control. To our knowledge, this is the first report that extracts of Cypreae moneta shell and Syzygium aromaticum flowering bud have tyrosinase inhibitory activity. These potent extracts were further evaluated at different concentration. The final concentration of the extracts in reaction mixtures was 50, 25 and 5 microg mL(-1) for the initial concentration of 1000, 500 and 100 microg mL(-1), respectively. They showed concentration-dependent inhibition of mushroom tyrosinase. Those extracts expressing relatively weak tyrosinase inhibitory activity may act through different inhibition pathway which is not based on tyrosinase activity. Further evaluation of the most potent tyrosinase inhibitors in in vivo conditions would be recommended.

  1. Synthesis and in Vitro Screening of Phenylbipyridinylpyrazole Derivatives as Potential Antiproliferative Agents

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    Mohammad M. Al-Sanea

    2015-01-01

    Full Text Available A series of phenylbipyridinylpyrazoles was synthesized through the reaction of 2-(4-(2-chloropyridin-4-yl-3-(3-methoxy-5-methylphenyl-1H-pyrazol-1-ylacetonitrile (4 with different 6-substituted pyridine-3-ylboronic acids. The final compounds 5a–j were screened at 10 µM against over 60 tumor cell lines at the U.S. National Cancer Institute (NCI. In light of the NCI results, compounds 5c and 5h showed a broad spectrum of activity against NCI cell lines with mean growth of 53% and 58%, respectively. Compound 5e behaved differently as it showed high degree of selectivity and potency by inhibiting 96% of growth of leukemia SR cell line at 10 µM. Standard COMPARE analyses were performed at the GI50 level and the results exhibit high correlation in the form of pairwise correlation coefficient (PCC of more than 0.6 between three of the current compounds and three standard known anticancer agents. Compound 5e demonstrated high correlation levels with merbarone (NSC S336628 with a PCC value of 0.631. Compound 5h showed a considerably high PCC value of 0.626 with dichloroallyl lawsone, while compound 5i, showed PCC values of 0.601 and 0.604 with both dichloroallyl lawsone and N,N-dibenzyldaunomycin (NSC S268242, respectively. These three standard agents have anticancer activity via two major mechanism of actions, inhibition of topoisomerase II and inhibition of biosynthesis of pyrimidine nucleotides, therefore, compounds 5a–j are promising therapeutic agents for targeting different human malignancies. Prediction of drug-likeness and toxicity of these newly synthesized derivatives were also considered.

  2. Development of an in vitro multicellular tumor spheroid model using microencapsulation and its application in anticancer drug screening and testing.

    Science.gov (United States)

    Zhang, Xulang; Wang, Wei; Yu, Weiting; Xie, Yubing; Zhang, Xiaohui; Zhang, Ying; Ma, Xiaojun

    2005-01-01

    In this study, an in vitro multicellular tumor spheroid model was developed using microencapsulation, and the feasibility of using the microencapsulated multicellular tumor spheroid (MMTS) to test the effect of chemotherapeutic drugs was investigated. Human MCF-7 breast cancer cells were encapsulated in alginate-poly-l-lysine-alginate (APA) microcapsules, and a single multicellular spheroid 150 mum in diameter was formed in the microcapsule after 5 days of cultivation. The cell morphology, proliferation, and viability of the MMTS were characterized using phase contrast microscopy, BrdU-labeling, MTT stain, calcein AM/ED-2 stain, and H&E stain. It demonstrated that the MMTS was viable and that the proliferating cells were mainly localized to the periphery of the cell spheroid and the apoptotic cells were in the core. The MCF-7 MMTS was treated with mitomycin C (MC) at a concentration of 0.1, 1, or 10 times that of peak plasma concentration (ppc) for up to 72 h. The cytotoxicity was demonstrated clearly by the reduction in cell spheroid size and the decrease in cell viability. The MMTS was further used to screen the anticancer effect of chemotherapeutic drugs, treated with MC, adriamycin (ADM) and 5-fluorouracil (5-FU) at concentrations of 0.1, 1, and 10 ppc for 24, 48, and 72 h. MCF-7 monolayer culture was used as control. Similar to monolayer culture, the cell viability of MMTS was reduced after treatment with anticancer drugs. However, the inhibition rate of cell viability in MMTS was much lower than that in monolayer culture. The MMTS was more resistant to anticancer drugs than monolayer culture. The inhibition rates of cell viability were 68.1%, 45.1%, and 46.8% in MMTS and 95.1%, 86.8%, and 91.6% in monolayer culture treated with MC, ADM, and 5-FU at 10 ppc for 72 h, respectively. MC showed the strongest cytotoxicity in both MMTS and monolayer, followed by 5-FU and ADM. It demonstrated that the MMTS has the potential to be a rapid and valid in vitro model to

  3. Repositioning FDA Drugs as Potential Cruzain Inhibitors from Trypanosoma cruzi: Virtual Screening, In Vitro and In Vivo Studies

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    Isidro Palos

    2017-06-01

    Full Text Available Chagas disease (CD is a neglected disease caused by the parasite Trypanosoma cruzi, which affects underdeveloped countries. The current drugs of choice are nifurtimox and benznidazole, but both have severe adverse effects and less effectivity in chronic infections; therefore, the need to discover new drugs is essential. A computer-guided drug repositioning method was applied to identify potential FDA drugs (approved and withdrawn as cruzain (Cz inhibitors and trypanocidal effects were confirmed by in vitro and in vivo studies. 3180 FDA drugs were virtually screened using a structure-based approach. From a first molecular docking analysis, a set of 33 compounds with the best binding energies were selected. Subsequent consensus affinity binding, ligand amino acid contact clustering analysis, and ranked position were used to choose four known pharmacological compounds to be tested in vitro. Mouse blood samples infected with trypomastigotes from INC-5 and NINOA strains were used to test the trypanocidal effect of four selected compounds. Among these drugs, one fibrate antilipemic (etofyllin clofibrate and three β-lactam antibiotics (piperacillin, cefoperazone, and flucloxacillin showed better trypanocidal effects (LC50 range 15.8–26.1 μg/mL in comparison with benznidazole and nifurtimox (LC50 range 33.1–46.7 μg/mL. A short-term in vivo evaluation of these compounds showed a reduction of parasitemia in infected mice (range 90–60% at 6 h, but this was low compared to benznidazole (50%. This work suggests that four known FDA drugs could be used to design and obtain new trypanocidal agents.

  4. Multifaceted role of nitric oxide in an in vitro mouse neuronal injury model: transcriptomic profiling defines the temporal recruitment of death signalling cascades

    Science.gov (United States)

    Peng, Zhao Feng; Chen, Minghui Jessica; Manikandan, Jayapal; Melendez, Alirio J; Shui, Guanghou; Russo-Marie, Françoise; Whiteman, Matthew; Beart, Philip M; Moore, Philip K; Cheung, Nam Sang

    2012-01-01

    Abstract Nitric oxide is implicated in the pathogenesis of various neuropathologies characterized by oxidative stress. Although nitric oxide has been reported to be involved in the exacerbation of oxidative stress observed in several neuropathologies, existent data fail to provide a holistic description of how nitrergic pathobiology elicits neuronal injury. Here we provide a comprehensive description of mechanisms contributing to nitric oxide induced neuronal injury by global transcriptomic profiling. Microarray analyses were undertaken on RNA from murine primary cortical neurons treated with the nitric oxide generator DETA-NONOate (NOC-18, 0.5 mM) for 8–24 hrs. Biological pathway analysis focused upon 3672 gene probes which demonstrated at least a ±1.5-fold expression in a minimum of one out of three time-points and passed statistical analysis (one-way anova, P < 0.05). Numerous enriched processes potentially determining nitric oxide mediated neuronal injury were identified from the transcriptomic profile: cell death, developmental growth and survival, cell cycle, calcium ion homeostasis, endoplasmic reticulum stress, oxidative stress, mitochondrial homeostasis, ubiquitin-mediated proteolysis, and GSH and nitric oxide metabolism. Our detailed time-course study of nitric oxide induced neuronal injury allowed us to provide the first time a holistic description of the temporal sequence of cellular events contributing to nitrergic injury. These data form a foundation for the development of screening platforms and define targets for intervention in nitric oxide neuropathologies where nitric oxide mediated injury is causative. PMID:21352476

  5. Induced Pluripotent Stem Cell Models to Enable In Vitro Models for Screening in the Central Nervous System

    Science.gov (United States)

    Efthymiou, Anastasia G.; Malik, Nasir; Behl, Mamta; Mead, Ivy L.; Zeng, Xianmin; Simeonov, Anton; Rao, Mahendra

    2015-01-01

    There is great need to develop more predictive drug discovery tools to identify new therapies to treat diseases of the central nervous system (CNS). Current nonpluripotent stem cell-based models often utilize non-CNS immortalized cell lines and do not enable the development of personalized models of disease. In this review, we discuss why in vitro models are necessary for translational research and outline the unique advantages of induced pluripotent stem cell (iPSC)-based models over those of current systems. We suggest that iPSC-based models can be patient specific and isogenic lines can be differentiated into many neural cell types for detailed comparisons. iPSC-derived cells can be combined to form small organoids, or large panels of lines can be developed that enable new forms of analysis. iPSC and embryonic stem cell-derived cells can be readily engineered to develop reporters for lineage studies or mechanism of action experiments further extending the utility of iPSC-based systems. We conclude by describing novel technologies that include strategies for the development of diversity panels, novel genomic engineering tools, new three-dimensional organoid systems, and modified high-content screens that may bring toxicology into the 21st century. The strategic integration of these technologies with the advantages of iPSC-derived cell technology, we believe, will be a paradigm shift for toxicology and drug discovery efforts. PMID:25794298

  6. In vitro antimicrobial and antiprotozoal activities, phytochemical screening and heavy metals toxicity of different parts of Ballota nigra.

    Science.gov (United States)

    Ullah, Najeeb; Ahmad, Ijaz; Ayaz, Sultan

    2014-01-01

    The study was done to assess the phytochemicals (flavonoids, terpenoids, saponins, tannin, alkaloids, and phenol) in different parts (root, stem, and leaves) of Ballota nigra and correlated it to inhibition of microbes (bacteria and fungi), protozoan (Leishmania), and heavy metals toxicity evaluation. In root and stem flavonoids, terpenes and phenols were present in ethanol, chloroform, and ethyl acetate soluble fraction; these were found to be the most active inhibiting fractions against all the tested strains of bacteria, fungi, and leishmania. While in leaves flavonoids, terpenes, and phenols were present in ethanol, chloroform, and n-butanol fractions which were the most active fractions against both types of microbes and protozoan (leishmania) in in vitro study. Ethanol and chloroform fractions show maximum inhibition against Escherichia coli (17 mm). The phytochemical and biological screenings were correlated with the presence of heavy metals in selected plant Ballota nigra. Cr was found above permissible value (above 1.5 mg/kg) in all parts of the plant. Ni was above WHO limit in B. nigra root and leaves (3.35 ± 1.20 mg/kg and 5.09 ± 0.47 mg/kg, respectively). Fe was above permissible value in all parts of B. nigra (above 20 mg/kg). Cd was above permissible value in all parts of the plant (above 0.3 mg/kg). Pb was above WHO limit (above 2 mg/kg) in all parts of Ballota nigra.

  7. Evaluation of OASIS QSAR Models Using ToxCast™ in Vitro Estrogen and Androgen Receptor Binding Data and Application in an Integrated Endocrine Screening Approach.

    Science.gov (United States)

    Bhhatarai, Barun; Wilson, Daniel M; Price, Paul S; Marty, Sue; Parks, Amanda K; Carney, Edward

    2016-09-01

    Integrative testing strategies (ITSs) for potential endocrine activity can use tiered in silico and in vitro models. Each component of an ITS should be thoroughly assessed. We used the data from three in vitro ToxCast™ binding assays to assess OASIS, a quantitative structure-activity relationship (QSAR) platform covering both estrogen receptor (ER) and androgen receptor (AR) binding. For stronger binders (described here as AC50 75%) and specificity (> 86%) for ER as well as both high sensitivity (92-100%) and specificity (70-81%) for AR. For compounds within the domains of the ER and AR QSAR models that bound with AC50 approach wherein a) QSAR is used to identify compounds in-domain of the ER or AR binding models and predicted to bind; b) those compounds are screened in vitro to assess binding potency; and c) the stronger binders (AC50 vitro, that require metabolism to manifest activity, or for which in vivo AR testing is in order, need to be assessed differently. Bhhatarai B, Wilson DM, Price PS, Marty S, Parks AK, Carney E. 2016. Evaluation of OASIS QSAR models using ToxCast™ in vitro estrogen and androgen receptor binding data and application in an integrated endocrine screening approach. Environ Health Perspect 124:1453-1461; http://dx.doi.org/10.1289/EHP184.

  8. New Imidazole Inhibitors of Mycobacterial FtsZ: the Way from High-Throughput Molecular Screening in Grid up to in vitro Verification

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    Karpov, P.A.

    2016-05-01

    Full Text Available In the framework of UNG virtual organization CSLabGrid, high-throughput molecular screening was performed for new anti-TB compounds. Using program FlexX installed on IFBG Claster and models of four perspective ligand binding sites on the surface of FtsZ of Mycobacterium tuberculosis, virtual screening was performed for database containing 2886 compounds synthesized in the Institute of Organic Chemistry of NAS of Ukraine. Based on LE and ΔG score, docking scores of CCDC Gold, and results of molecular dynamics, we selected a group of perspective FtsZ inhibitors. In vitro validation have revealed 6 compounds with the highest inhibition of GTPase activity of FtsZ. Also, based on in vitro experiment, we have selected three compounds exhibiting both - strong inhibition of FtsZ polymerization and inhibition of GTPase activity.

  9. In vitro antioxidant activity, phytochemical screening, cytotoxicity and total phenolic content in extracts of Caesalpinia pulcherrima (Caesalpiniaceae) pods.

    Science.gov (United States)

    Kumbhare, M R; Sivakumar, T; Udavant, P B; Dhake, A S; Surana, A R

    2012-04-01

    Caesalpinia pulcherrima L. Swartz (Caesalpiniaceae) is an ornamental plant also used as a common medicinal plant in India, Taiwan and South-East Asian countries. Majority of the diseases/disorders are mainly linked to oxidative stress due to free radicals. The aims of this study were to screen for phytochemical constituents, evaluate cytotoxicity, in vitro antioxidant activity and estimation of total phenolic content of extracts of pods of Caesalpinia pulcherrima. Phytochemical analysis revealed the presence of tannins, flavonoids, steroids and alkaloids. Brine Shrimp Lethality (BSL) bioassay was used to investigate the cytotoxic effects. The LC50(microg mL(-1)) values obtained for extracts as 750 microg mL(-1) for petroleum ether extract, 800 microg mL(-1) for chloroform extract and 900 microg mL(-1) for methanol extract. The total phenolic content of the methanolic extract was 38.04% w/w, equivalent to gallic acid. Petroleum ether, chloroform and methanolic extracts of Caesalpinia pulcherrima and standard ascorbic acid were found to be scavenger of DPPH radical with an IC50 of 124.75, 112.08, 54.34 and 13.86 microg mL(-1), respectively. Methanolic extract was good scavenger of DPPH radical. Petroleum ether, chloroform, ethyl acetate soluble fraction of methanolic extracts of pods of Caesalpinia pulcherrima and ascorbic acid were found to be scavenger of nitric oxide radical with an IC50 of 93.32, 65.12, 54.83 and 12.59 microg mL(-1), respectively. Ethyl acetate soluble fraction was found to be good scavenger of nitric oxide radical. Our conclusion provides support that the crude extracts of C. pulcherrima is a probable source of natural antioxidants and this justified its uses in folkloric medicines.

  10. In Vitro Antimicrobial and Antiprotozoal Activities, Phytochemical Screening and Heavy Metals Toxicity of Different Parts of Ballota nigra

    Directory of Open Access Journals (Sweden)

    Najeeb Ullah

    2014-01-01

    Full Text Available The study was done to assess the phytochemicals (flavonoids, terpenoids, saponins, tannin, alkaloids, and phenol in different parts (root, stem, and leaves of Ballota nigra and correlated it to inhibition of microbes (bacteria and fungi, protozoan (Leishmania, and heavy metals toxicity evaluation. In root and stem flavonoids, terpenes and phenols were present in ethanol, chloroform, and ethyl acetate soluble fraction; these were found to be the most active inhibiting fractions against all the tested strains of bacteria, fungi, and leishmania. While in leaves flavonoids, terpenes, and phenols were present in ethanol, chloroform, and n-butanol fractions which were the most active fractions against both types of microbes and protozoan (leishmania in in vitro study. Ethanol and chloroform fractions show maximum inhibition against Escherichia coli (17 mm. The phytochemical and biological screenings were correlated with the presence of heavy metals in selected plant Ballota nigra. Cr was found above permissible value (above 1.5 mg/kg in all parts of the plant. Ni was above WHO limit in B. nigra root and leaves (3.35 ± 1.20 mg/kg and 5.09 ± 0.47 mg/kg, respectively. Fe was above permissible value in all parts of B. nigra (above 20 mg/kg. Cd was above permissible value in all parts of the plant (above 0.3 mg/kg. Pb was above WHO limit (above 2 mg/kg in all parts of Ballota nigra.

  11. In vitro screening for protein tyrosine phosphatase 1B and dipeptidyl peptidase IV inhibitors from selected Nigerian medicinal plants.

    Science.gov (United States)

    Saidu, Yusuf; Muhammad, Suleiman Alhaji; Abbas, Abdullahi Yahaya; Onu, Andrew; Tsado, Ibrahim Mohammed; Muhammad, Luba

    2017-01-01

    Protein tyrosine phosphatase 1B (PTP 1B) and dipeptidyl peptidase IV (DPP IV) have been identified as one of the drug targets for the treatment of Type-2 diabetes. This study was designed to screen for PTP 1B and DPP-IV inhibitors from some Nigerian medicinal plants. PTP 1B and DPP-IV drug discovery kits from Enzo Life Sciences were used to investigate in vitro inhibitory effect of crude methanolic extract of 10 plants; Mangifera indica, Moringa oleifera, Acacia nilotica, Arachis hypogaea, Senna nigricans, Azadirachta indica, Calotropis procera, Leptadenia hastata, Ziziphus mauritiana, and Solanum incanum. The results indicated PTP IB inhibition by S. nigricans (68.2 ± 2.29%), A. indica (67.4 ± 2.80%), A. hypogaea (57.2 ± 2.50%), A. nilotica (55.1 ± 2.19%), and M. oleifera (41.2 ± 1.87%) were significantly (P 0.05) different from that of sumarin. The DPP-IV inhibition by S. incanum (68.1 ± 2.71%) was significantly higher as compared with a known inhibitor, P32/98. S. nigrican (57.0±1.91%), Z. mauritiana (56.6±2.01%), A. hypogaea (51.0±1.30%), M. indica (44.6 ± 2.40%), C. procera (36.2 ± 2.00%), A. nilotica (35.4 ± 2.10%), and A. indica (33.6 ± 1.50%) show significantly (P < 0.05) lower inhibitions toward DPP-IV. The work demonstrated that these plant materials could serve as sources of lead compounds in the development of anti-diabetic agent(s) targeting PTP 1B and/or DPP-IV.

  12. Computationally derived points of fragility of a human cascade are consistent with current therapeutic strategies.

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    Deyan Luan

    2007-07-01

    Full Text Available The role that mechanistic mathematical modeling and systems biology will play in molecular medicine and clinical development remains uncertain. In this study, mathematical modeling and sensitivity analysis were used to explore the working hypothesis that mechanistic models of human cascades, despite model uncertainty, can be computationally screened for points of fragility, and that these sensitive mechanisms could serve as therapeutic targets. We tested our working hypothesis by screening a model of the well-studied coagulation cascade, developed and validated from literature. The predicted sensitive mechanisms were then compared with the treatment literature. The model, composed of 92 proteins and 148 protein-protein interactions, was validated using 21 published datasets generated from two different quiescent in vitro coagulation models. Simulated platelet activation and thrombin generation profiles in the presence and absence of natural anticoagulants were consistent with measured values, with a mean correlation of 0.87 across all trials. Overall state sensitivity coefficients, which measure the robustness or fragility of a given mechanism, were calculated using a Monte Carlo strategy. In the absence of anticoagulants, fluid and surface phase factor X/activated factor X (fX/FXa activity and thrombin-mediated platelet activation were found to be fragile, while fIX/FIXa and fVIII/FVIIIa activation and activity were robust. Both anti-fX/FXa and direct thrombin inhibitors are important classes of anticoagulants; for example, anti-fX/FXa inhibitors have FDA approval for the prevention of venous thromboembolism following surgical intervention and as an initial treatment for deep venous thrombosis and pulmonary embolism. Both in vitro and in vivo experimental evidence is reviewed supporting the prediction that fIX/FIXa activity is robust. When taken together, these results support our working hypothesis that computationally derived points of

  13. Salicylic and jasmonic acid pathways are necessary for defence against Dickeya solani as revealed by a novel method for Blackleg disease screening of in vitro grown potato.

    Science.gov (United States)

    Burra, D D; Mühlenbock, P; Andreasson, E

    2015-09-01

    Potato is major crop ensuring food security in Europe, and blackleg disease is increasingly causing losses in yield and during storage. Recently, one blackleg pathogen, Dickeya solani has been shown to be spreading in Northern Europe that causes aggressive disease development. Currently, identification of tolerant commercial potato varieties has been unsuccessful; this is confounded by the complicated etiology of the disease and a strong environmental influence on disease development. There is currently a lack of efficient testing systems. Here, we describe a system for quantification of blackleg symptoms on shoots of sterile in vitro potato plants, which saves time and space compared to greenhouse and existing field assays. We found no evidence for differences in infection between the described in vitro-based screening method and existing greenhouse assays. This system facilitates efficient screening of blackleg disease response of potato plants independent of other microorganisms and variable environmental conditions. We therefore used the in vitro screening method to increase understanding of plant mechanisms involved in blackleg disease development by analysing disease response of hormone- related (salicylic and jasmonic acid) transgenic potato plants. We show that both jasmonic (JA) and salicylic (SA) acid pathways regulate tolerance to blackleg disease in potato, a result unlike previous findings in Arabidopsis defence response to necrotrophic bacteria. We confirm this by showing induction of a SA marker, pathogenesis-related protein 1 (StPR1), and a JA marker, lipoxygenase (StLOX), in Dickeya solani infected in vitro potato plants. We also observed that tubers of transgenic potato plants were more susceptible to soft rot compared to wild type, suggesting a role for SA and JA pathways in general tolerance to Dickeya. © 2015 German Botanical Society and The Royal Botanical Society of the Netherlands.

  14. Application of an in vitro drug screening assay based on the release of phosphoglucose isomerase to determine the structure-activity relationship of thiazolides against Echinococcus multilocularis metacestodes.

    Science.gov (United States)

    Stadelmann, Britta; Scholl, Sabrina; Müller, Joachim; Hemphill, Andrew

    2010-03-01

    The disease alveolar echinococcosis (AE), caused by the larval stage of the cestode Echinococcus multilocularis, is fatal if treatment is unsuccessful. Current treatment options are, at best, parasitostatic, and involve taking benzimidazoles (albendazole, mebendazole) for the whole of a patient's life. In conjunction with the recent development of optimized procedures for E. multilocularis metacestode cultivation, we aimed to develop a rapid and reliable drug screening test, which enables efficient screening of a large number of compounds in a relatively short time frame. Metacestodes were treated in vitro with albendazole, the nitro-thiazole nitazoxanide and 29 nitazoxanide derivatives. The resulting leakage of phosphoglucose isomerase (PGI) activity into the medium supernatant was measured and provided an indication of compound efficacy. We show that upon in vitro culture of E. multilocularis metacestodes in the presence of active drugs such as albendazole, the nitro-thiazole nitazoxanide and 30 different nitazoxanide derivatives, the activity of PGI in culture supernatants increased. The increase in PGI activity correlated with the progressive degeneration and destruction of metacestode tissue in a time- and concentration-dependent manner, which allowed us to perform a structure-activity relationship analysis on the thiazolide compounds used in this study. The assay presented here is inexpensive, rapid, can be used in 24- and 96-well formats and will serve as an ideal tool for first-round in vitro tests on the efficacy of large numbers of antiparasitic compounds.

  15. Identification of hepta-histidine as a candidate drug for Huntington’s disease by in silico-in vitro- in vivo-integrated screens of chemical libraries

    Science.gov (United States)

    Imamura, Tomomi; Fujita, Kyota; Tagawa, Kazuhiko; Ikura, Teikichi; Chen, Xigui; Homma, Hidenori; Tamura, Takuya; Mao, Ying; Taniguchi, Juliana Bosso; Motoki, Kazumi; Nakabayashi, Makoto; Ito, Nobutoshi; Yamada, Kazunori; Tomii, Kentaro; Okano, Hideyuki; Kaye, Julia; Finkbeiner, Steven; Okazawa, Hitoshi

    2016-09-01

    We identified drug seeds for treating Huntington’s disease (HD) by combining in vitro single molecule fluorescence spectroscopy, in silico molecular docking simulations, and in vivo fly and mouse HD models to screen for inhibitors of abnormal interactions between mutant Htt and physiological Ku70, an essential DNA damage repair protein in neurons whose function is known to be impaired by mutant Htt. From 19,468 and 3,010,321 chemicals in actual and virtual libraries, fifty-six chemicals were selected from combined in vitro-in silico screens; six of these were further confirmed to have an in vivo effect on lifespan in a fly HD model, and two chemicals exerted an in vivo effect on the lifespan, body weight and motor function in a mouse HD model. Two oligopeptides, hepta-histidine (7H) and Angiotensin III, rescued the morphological abnormalities of primary neurons differentiated from iPS cells of human HD patients. For these selected drug seeds, we proposed a possible common structure. Unexpectedly, the selected chemicals enhanced rather than inhibited Htt aggregation, as indicated by dynamic light scattering analysis. Taken together, these integrated screens revealed a new pathway for the molecular targeted therapy of HD.

  16. Cascade Organic Solar Cells

    KAUST Repository

    Schlenker, Cody W.

    2011-09-27

    We demonstrate planar organic solar cells consisting of a series of complementary donor materials with cascading exciton energies, incorporated in the following structure: glass/indium-tin-oxide/donor cascade/C 60/bathocuproine/Al. Using a tetracene layer grown in a descending energy cascade on 5,6-diphenyl-tetracene and capped with 5,6,11,12-tetraphenyl- tetracene, where the accessibility of the π-system in each material is expected to influence the rate of parasitic carrier leakage and charge recombination at the donor/acceptor interface, we observe an increase in open circuit voltage (Voc) of approximately 40% (corresponding to a change of +200 mV) compared to that of a single tetracene donor. Little change is observed in other parameters such as fill factor and short circuit current density (FF = 0.50 ± 0.02 and Jsc = 2.55 ± 0.23 mA/cm2) compared to those of the control tetracene-C60 solar cells (FF = 0.54 ± 0.02 and Jsc = 2.86 ± 0.23 mA/cm2). We demonstrate that this cascade architecture is effective in reducing losses due to polaron pair recombination at donor-acceptor interfaces, while enhancing spectral coverage, resulting in a substantial increase in the power conversion efficiency for cascade organic photovoltaic cells compared to tetracene and pentacene based devices with a single donor layer. © 2011 American Chemical Society.

  17. Adaption of an in vitro digestion method to screen carotenoid liberation and in vitro accessibility from differently processed spinach preparations

    DEFF Research Database (Denmark)

    Eriksen, Jane Nygaard; Luu, Amy Y; Dragsted, Lars Ove

    2017-01-01

    Dark green leafy vegetables are primary food sources for lutein and β-carotene, however these bioactives have low bioavailability. The effects of mechanical and thermal processing as well as fat addition and fat type on lutein and β-carotene liberation and in vitro accessibility from spinach were...

  18. Digestion of wood-based hemicellulose extracts as screened by in vitro gas production method and verified in vivo using sheep

    Directory of Open Access Journals (Sweden)

    Marketta Rinne

    2016-03-01

    Full Text Available The objective of the present study was to evaluate the potential of pressurized hot water extracted hemicellulose fractions from various wood species as feeds for ruminants. In Experiment 1, the fermentability of several hemicellulose extracts was screened using an in vitro gas production method. The samples were extracted from spruce (Picea abies including mainly galactoglucomannan (GGM, from birch (Betula pendula consisting mainly of xylan and from larch (Larix sibirica consisting mainly of arabinogalactan. The GGM and xylan samples were readily fermented by rumen microbes while arabinogalactan was not. Based on the in vitro study, GGM was chosen for an in vivo digestibility trial using sheep, where it was fed at increasing proportions of diet dry matter (0, 47, 94 and 141 g kg-1 in a Latin Square design. The in vivo organic matter digestibility of GGM was relatively low, 0.58, but PHWE extracted hemicellulose has some potential as a feed for ruminants.

  19. CIRCLE-seq: a highly sensitive in vitro screen for genome-wide CRISPR-Cas9 nuclease off-targets.

    Science.gov (United States)

    Tsai, Shengdar Q; Nguyen, Nhu T; Malagon-Lopez, Jose; Topkar, Ved V; Aryee, Martin J; Joung, J Keith

    2017-06-01

    Sensitive detection of off-target effects is important for translating CRISPR-Cas9 nucleases into human therapeutics. In vitro biochemical methods for finding off-targets offer the potential advantages of greater reproducibility and scalability while avoiding limitations associated with strategies that require the culture and manipulation of living cells. Here we describe circularization for in vitro reporting of cleavage effects by sequencing (CIRCLE-seq), a highly sensitive, sequencing-efficient in vitro screening strategy that outperforms existing cell-based or biochemical approaches for identifying CRISPR-Cas9 genome-wide off-target mutations. In contrast to previously described in vitro methods, we show that CIRCLE-seq can be practiced using widely accessible next-generation sequencing technology and does not require reference genome sequences. Importantly, CIRCLE-seq can be used to identify off-target mutations associated with cell-type-specific single-nucleotide polymorphisms, demonstrating the feasibility and importance of generating personalized specificity profiles. CIRCLE-seq provides an accessible, rapid, and comprehensive method for identifying genome-wide off-target mutations of CRISPR-Cas9.

  20. Elimination of teicoplanin by adsorption to the filter membrane during haemodiafiltration: screening experiments for linezolid, teicoplanin and vancomycin followed by in vitro haemodiafiltration models for teicoplanin.

    Science.gov (United States)

    Shiraishi, Y; Okajima, M; Sai, Y; Miyamoto, K; Inaba, H

    2012-05-01

    Pharmaceutical agents directed against methicillin-resistant Staphylococcus aureus can be eliminated during haemodiafiltration, not only by diffusion and ultrafiltration, but also by adsorption onto haemofilters. The latter may be affected by the binding of agents to serum albumin. The present study therefore investigated the affinity of anti-methicillin-resistant Staphylococcus aureus agents (teicoplanin, linezolid, vancomycin) for haemofilters and the pharmacokinetic properties of teicoplanin during haemodiafiltration. Linezolid, teicoplanin and vancomycin were first screened for their in vitro affinity for three different kinds of filter membranes: polysulfone, polyacrylonitrile and polymethylmethacrylate. Only teicoplanin showed significant filter-binding activity. An in vitro haemodiafiltration circulation model was then developed that incorporated a one-litre beaker containing Krebs-Ringer's bicarbonate solution with/without human albumin (0 or 3 g/dl) as an artificial plasma. Teicoplanin (initial concentration 50 µg/ml, representing the maximum plasma concentration (Cmax) resulting from a typical clinical dosage) was circulated throughout the beaker. Teicoplanin concentrations in the 'plasma' and ultrafiltrate were determined by high performance liquid chromatography. In the screening experiment, teicoplanin was predominantly adsorbed onto polysulfone and polymethylmethacrylate membranes. Furthermore, teicoplanin was primarily eliminated by adsorption onto these filters during in vitro haemodiafiltration. Albumin significantly reduced both haemodiafiltration clearance and the adsorption-dependent elimination, although there were complex but significant interactions between albumin and the filter membrane. Elimination of teicoplanin in an in vitro haemodiafiltration model was largely due to adsorption onto polysulfone and polymethylmethacrylate haemofilters. Future clinical studies should likely be designed to evaluate present recommendations of teicoplanin

  1. Howling about Trophic Cascades

    Science.gov (United States)

    Kowalewski, David

    2012-01-01

    Following evolutionary theory and an agriculture model, ecosystem research has stressed bottom-up dynamics, implying that top wild predators are epiphenomenal effects of more basic causes. As such, they are assumed expendable. A more modern co-evolutionary and wilderness approach--trophic cascades--instead suggests that top predators, whose…

  2. Establishing in vitro in vivo correlations to screen monoclonal antibodies for physicochemical properties related to favorable human pharmacokinetics.

    Science.gov (United States)

    Avery, Lindsay B; Wade, Jason; Wang, Mengmeng; Tam, Amy; King, Amy; Piche-Nicholas, Nicole; Kavosi, Mania S; Penn, Steve; Cirelli, David; Kurz, Jeffrey C; Zhang, Minlei; Cunningham, Orla; Jones, Rhys; Fennell, Brian J; McDonnell, Barry; Sakorafas, Paul; Apgar, James; Finlay, William J; Lin, Laura; Bloom, Laird; O'Hara, Denise M

    2017-12-22

    Implementation of in vitro assays that correlate with in vivo human pharmacokinetics (PK) would provide desirable preclinical tools for the early selection of therapeutic monoclonal antibody (mAb) candidates with minimal non-target-related PK risk. Use of these tools minimizes the likelihood that mAbs with unfavorable PK would be advanced into costly preclinical and clinical development. In total, 42 mAbs varying in isotype and soluble versus membrane targets were tested in in vitro and in vivo studies. MAb physicochemical properties were assessed by measuring non-specific interactions (DNA- and insulin-binding ELISA), self-association (affinity-capture self-interaction nanoparticle spectroscopy) and binding to matrix-immobilized human FcRn (surface plasmon resonance and column chromatography). The range of scores obtained from each in vitro assay trended well with in vivo clearance (CL) using both human FcRn transgenic (Tg32) mouse allometrically projected human CL and observed human CL, where mAbs with high in vitro scores resulted in rapid CL in vivo. Establishing a threshold value for mAb CL in human of 0.32 mL/hr/kg enabled refinement of thresholds for each in vitro assay parameter, and using a combinatorial triage approach enabled the successful differentiation of mAbs at high risk for rapid CL (unfavorable PK) from those with low risk (favorable PK), which allowed mAbs requiring further characterization to be identified. Correlating in vitro parameters with in vivo human CL resulted in a set of in vitro tools for use in early testing that would enable selection of mAbs with the greatest likelihood of success in the clinic, allowing costly late-stage failures related to an inadequate exposure profile, toxicity or lack of efficacy to be avoided.

  3. In Vitro Tumor Models: Advantages, Disadvantages, Variables, and Selecting the Right Platform.

    Science.gov (United States)

    Katt, Moriah E; Placone, Amanda L; Wong, Andrew D; Xu, Zinnia S; Searson, Peter C

    2016-01-01

    In vitro tumor models have provided important tools for cancer research and serve as low-cost screening platforms for drug therapies; however, cancer recurrence remains largely unchecked due to metastasis, which is the cause of the majority of cancer-related deaths. The need for an improved understanding of the progression and treatment of cancer has pushed for increased accuracy and physiological relevance of in vitro tumor models. As a result, in vitro tumor models have concurrently increased in complexity and their output parameters further diversified, since these models have progressed beyond simple proliferation, invasion, and cytotoxicity screens and have begun recapitulating critical steps in the metastatic cascade, such as intravasation, extravasation, angiogenesis, matrix remodeling, and tumor cell dormancy. Advances in tumor cell biology, 3D cell culture, tissue engineering, biomaterials, microfabrication, and microfluidics have enabled rapid development of new in vitro tumor models that often incorporate multiple cell types, extracellular matrix materials, and spatial and temporal introduction of soluble factors. Other innovations include the incorporation of perfusable microvessels to simulate the tumor vasculature and model intravasation and extravasation. The drive toward precision medicine has increased interest in adapting in vitro tumor models for patient-specific therapies, clinical management, and assessment of metastatic potential. Here, we review the wide range of current in vitro tumor models and summarize their advantages, disadvantages, and suitability in modeling specific aspects of the metastatic cascade and drug treatment.

  4. Probe molecule (PrM) approach in adverse outcome pathway (AOP) based high-throughput screening (HTS): in vivo discovery for developing in vitro target methods.

    Science.gov (United States)

    Angrish, Michelle M; Madden, Michael C; Pleil, Joachim D

    2015-04-20

    Efficient and accurate adverse outcome pathway (AOP) based high-throughput screening (HTS) methods use a systems biology based approach to computationally model in vitro cellular and molecular data for rapid chemical prioritization; however, not all HTS assays are grounded by relevant in vivo exposure data. The challenge is to develop HTS assays with unambiguous quantitative links between in vitro responses and corresponding in vivo effects, which is complicated by metabolically insufficient systems, in vitro to in vivo extrapolation (IVIVE), cross-species comparisons, and other inherent issues correlating IVIVE findings. This article introduces the concept of ultrasensitive gas phase probe molecules (PrMs) to help bridge the current HTS assay IVIVE gap. The PrM concept assesses metabolic pathways that have already been well-defined from intact human or mammalian models. Specifically, the idea is to introduce a gas phase probe molecule into a system, observe normal steady state, add chemicals of interest, and quantitatively measure (from headspace gas) effects on PrM metabolism that can be directly linked back to a well-defined and corresponding in vivo effect. As an example, we developed the pharmacokinetic (PK) parameters and differential equations to estimate methyl tertiary butyl ether (MTBE) metabolism to tertiary butyl alcohol (TBA) via cytochrome (CYP) 2A6 in the liver from human empirical data. Because MTBE metabolic pathways are well characterized from in vivo data, we can use it as a PrM to explore direct and indirect chemical effects on CYP pathways. The PrM concept could be easily applied to in vitro and alternative models of disease and phenotype, and even test for volatile chemicals while avoiding liquid handling robotics. Furthermore, a PrM can be designed for any chemical with known empirical human exposure data and used to assess chemicals for which no information exists. Herein, we propose an elegant gas phase probe molecule-based approach to in

  5. High-Throughput and Rapid Screening of Novel ACE Inhibitory Peptides from Sericin Source and Inhibition Mechanism by Using in Silico and in Vitro Prescriptions.

    Science.gov (United States)

    Sun, Huaju; Chang, Qing; Liu, Long; Chai, Kungang; Lin, Guangyan; Huo, Qingling; Zhao, Zhenxia; Zhao, Zhongxing

    2017-11-22

    Several novel peptides with high ACE-I inhibitory activity were successfully screened from sericin hydrolysate (SH) by coupling in silico and in vitro approaches for the first time. Most screening processes for ACE-I inhibitory peptides were achieved through high-throughput in silico simulation followed by in vitro verification. QSAR model based predicted results indicated that the ACE-I inhibitory activity of these SH peptides and six chosen peptides exhibited moderate high ACE-I inhibitory activities (log IC50 values: 1.63-2.34). Moreover, two tripeptides among the chosen six peptides were selected for ACE-I inhibition mechanism analysis which based on Lineweaver-Burk plots indicated that they behave as competitive ACE-I inhibitors. The C-terminal residues of short-chain peptides that contain more H-bond acceptor groups could easily form hydrogen bonds with ACE-I and have higher ACE-I inhibitory activity. Overall, sericin protein as a strong ACE-I inhibition source could be deemed a promising agent for antihypertension applications.

  6. Quantitative RT-PCR assay for high-throughput screening (HTS of drugs against the growth of Cryptosporidium parvum in vitro

    Directory of Open Access Journals (Sweden)

    Haili eZhang

    2015-09-01

    Full Text Available Our laboratory has previously developed a qRT-PCR assay to assess drug efficacy on the growth of Cryptosporidium parvum in vitro by detecting the levels of parasite 18S rRNA. This approach displayed up to four orders of magnitude of linear dynamic range and was much less labor-intensive than the traditional microscopic methods. However, conventional qRT-PCR protocol is not very amendable to high-throughput analysis when total RNA needs to be purified by lengthy, multi-step procedures. Recently, several commercial reagents are available for preparing cell lysates that could be directly used in downstream qRT-PCR analysis (e.g., Ambion Cell-to-cDNA kit and Bio-Rad iScript sample preparation reagent. Using these reagents, we are able to adapt the qRT-PCR assay into high-throughput screening of drugs in vitro (i.e., 96-well and 384-well formats for the cultivation of parasites and qRT-PCR detection, respectively. This qRT-PCR protocol is able to give a >150-fold linear dynamic range using samples isolated from cells infected with various numbers of parasites. The new assay is also validated by the NIH-recommended intra-plate, inter-plate and inter-day uniformity tests. The robustness and effectiveness of the assay are also confirmed by evaluating the anti-cryptosporidial efficacy of paromomycin and by a small scale screening of compounds.

  7. In vitro screening of Chinese medicinal toothpastes: their effects on growth and plaque formation of mutans streptococci.

    Science.gov (United States)

    Wu-Yuan, C D; Green, L; Birch, W X

    1990-01-01

    Many plant extracts or derivatives have been incorporated into commercial toothpastes to treat oral diseases related to caries or periodontal diseases in China. However, no information is available concerning their in vitro effects on oral bacteria. Thirty-one Chinese medicinal toothpastes were selected for this study. Their ability to inhibit growth, in vitro plaque formation and glucosyltransferase (GTF) activity of Streptococcus sobrinus B13 and Streptococcus mutans 3209 were examined. Eighty-seven percent of the tested toothpastes inhibited the growth of the mutans streptococci, with zones ranging from 0.8 to 2.5 cm. At 10 mg/ml, 74% of the toothpastes inhibited in vitro plaque formation by S. mutans. Among these, 60% completely suppressed water-insoluble glucan synthesis from sucrose by GTF. Based on data obtained from our study, the incorporation of natural plant products or their derivatives into dentifrices seems a feasible means of promoting oral health and controlling dental diseases.

  8. Risk-based high-throughput chemical screening and prioritization using exposure models and in vitro bioactivity assays

    DEFF Research Database (Denmark)

    Shin, Hyeong-Moo; Ernstoff, Alexi; Arnot, Jon

    2015-01-01

    relevant use scenarios (e.g., dermal application, indoor emissions). For each chemical and use scenario, exposure models are then used to calculate a chemical intake fraction, or a product intake fraction, accounting for chemical properties and the exposed population. We then combine these intake fractions...... with use scenario-specific estimates of chemical quantity to calculate daily intake rates (iR; mg/kg/day). These intake rates are compared to oral equivalent doses (OED; mg/kg/day), calculated from a suite of ToxCast in vitro bioactivity assays using in vitro-to-in vivo extrapolation and reverse dosimetry...

  9. Phytochemical screening, anti-oxidant activity and in vitro anticancer potential of ethanolic and water leaves extracts of Annona muricata (Graviola).

    Science.gov (United States)

    Gavamukulya, Yahaya; Abou-Elella, Faten; Wamunyokoli, Fred; AEl-Shemy, Hany

    2014-09-01

    To determine the phytochemical composition, antioxidant and anticancer activities of ethanolic and water leaves extracts of Annona muricata (A. muricata) from the Eastern Uganda. Phytochemical screening was conducted using standard qualitative methods and a Chi-square goodness of fit test was used to assign the relative abundance of the different phytochemicals. The antioxidant activity was determined using the 2, 2-diphenyl-2-picrylhydrazyl and reducing power methods whereas the in vitro anticancer activity was determined using three different cell lines. Phytochemical screening of the extracts revealed that they were rich in secondary class metabolite compounds such as alkaloids, saponins, terpenoids, flavonoids, coumarins and lactones, anthraquinones, tannins, cardiac glycosides, phenols and phytosterols. Total phenolics in the water extract were (683.69±0.09) μg/mL gallic acid equivalents (GAE) while it was (372.92±0.15) μg/mL GAE in the ethanolic extract. The reducing power was 216.41 μg/mL in the water extract and 470.51 μg/mL GAE in the ethanolic extract. In vitro antioxidant activity IC50 was 2.0456 mg/mL and 0.9077 mg/mL for ethanolic and water leaves extracts of A. muricata respectively. The ethanolic leaves extract was found to be selectively cytotoxic in vitro to tumor cell lines (EACC, MDA and SKBR3) with IC50 values of 335.85 μg/mL, 248.77 μg/mL, 202.33 μg/mL respectively, while it had no cytotoxic effect on normal spleen cells. The data also showed that water leaves extract of A. muricata had no anticancer effect at all tested concentrations. The results showed that A. muricata was a promising new antioxidant and anticancer agent. Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  10. A Workflow to Investigate Exposure and Pharmacokinetic Influences on High-Throughput in Vitro Chemical Screening Based on Adverse Outcome Pathways.

    Science.gov (United States)

    Phillips, Martin B; Leonard, Jeremy A; Grulke, Christopher M; Chang, Daniel T; Edwards, Stephen W; Brooks, Raina; Goldsmith, Michael-Rock; El-Masri, Hisham; Tan, Yu-Mei

    2016-01-01

    Adverse outcome pathways (AOPs) link adverse effects in individuals or populations to a molecular initiating event (MIE) that can be quantified using in vitro methods. Practical application of AOPs in chemical-specific risk assessment requires incorporation of knowledge on exposure, along with absorption, distribution, metabolism, and excretion (ADME) properties of chemicals. We developed a conceptual workflow to examine exposure and ADME properties in relation to an MIE. The utility of this workflow was evaluated using a previously established AOP, acetylcholinesterase (AChE) inhibition. Thirty chemicals found to inhibit human AChE in the ToxCast™ assay were examined with respect to their exposure, absorption potential, and ability to cross the blood-brain barrier (BBB). Structures of active chemicals were compared against structures of 1,029 inactive chemicals to detect possible parent compounds that might have active metabolites. Application of the workflow screened 10 "low-priority" chemicals of 30 active chemicals. Fifty-two of the 1,029 inactive chemicals exhibited a similarity threshold of ≥ 75% with their nearest active neighbors. Of these 52 compounds, 30 were excluded due to poor absorption or distribution. The remaining 22 compounds may inhibit AChE in vivo either directly or as a result of metabolic activation. The incorporation of exposure and ADME properties into the conceptual workflow eliminated 10 "low-priority" chemicals that may otherwise have undergone additional, resource-consuming analyses. Our workflow also increased confidence in interpretation of in vitro results by identifying possible "false negatives." Phillips MB, Leonard JA, Grulke CM, Chang DT, Edwards SW, Brooks R, Goldsmith MR, El-Masri H, Tan YM. 2016. A workflow to investigate exposure and pharmacokinetic influences on high-throughput in vitro chemical screening based on adverse outcome pathways. Environ Health Perspect 124:53-60; http://dx.doi.org/10.1289/ehp.1409450.

  11. In Silico Screening, Synthesis and In Vitro Evaluation of Some Quinazolinone and Pyridine Derivatives as Dihydrofolate Reductase Inhibitors for Anticancer Activity

    Directory of Open Access Journals (Sweden)

    A. G. Nerkar

    2009-01-01

    Full Text Available Dihydrofolate reductase (DHFR is the important target for anticancer drugs belonging to the class of antimetabolites as the enzyme plays important role in the de novo purine synthesis. We here report the in silico screening to obtain best fit molecules as DHFR inhibitors, synthesis of some ʻbest fitʼ quinazolinone from 2-phenyl-3-(substituted-benzilidine-amino quinazolinones (Quinazolinone Shiff's bases QSB1-5 and pyridine-4-carbohydrazide Shiff's bases (ISB1-5 derivatives and their in vitro anticancer assay. Synthesis of the molecules was performed using microwave assisted synthesis. The structures of these molecules were elucidated by IR and 1H-NMR. These compounds were then subjected for in vitro anticancer evaluation against five human cancer cell-lines for anticancer cyto-toxicity assay. Methotrexate (MTX was used as standard for this evaluation to give a comparable inhibition of the cell proliferation by DHFR inhibition. Placlitaxel, adriamycin and 5-fluoro-uracil were also used as standard to give a comparable activity of these compounds with other mechanism of anticancer activity. ISB3 (4-(N, N-dimethyl-amino-phenyl Schiff''s base derivative of pyridine carbohydrazide showed equipotent activity with the standards used in in vitro anticancer assay as per the NCI (National Cancer Institute guidelines.

  12. iPSC-Based Compound Screening and In Vitro Trials Identify a Synergistic Anti-amyloid β Combination for Alzheimer’s Disease

    Directory of Open Access Journals (Sweden)

    Takayuki Kondo

    2017-11-01

    Full Text Available In the process of drug development, in vitro studies do not always adequately predict human-specific drug responsiveness in clinical trials. Here, we applied the advantage of human iPSC-derived neurons, which offer human-specific drug responsiveness, to screen and evaluate therapeutic candidates for Alzheimer’s disease (AD. Using AD patient neurons with nearly 100% purity from iPSCs, we established a robust and reproducible assay for amyloid β peptide (Aβ, a pathogenic molecule in AD, and screened a pharmaceutical compound library. We acquired 27 Aβ-lowering screen hits, prioritized hits by chemical structure-based clustering, and selected 6 leading compounds. Next, to maximize the anti-Aβ effect, we selected a synergistic combination of bromocriptine, cromolyn, and topiramate as an anti-Aβ cocktail. Finally, using neurons from familial and sporadic AD patients, we found that the cocktail showed a significant and potent anti-Aβ effect on patient cells. This human iPSC-based platform promises to be useful for AD drug development.

  13. Comparison of two in vitro dendritic cell maturation models for screening contact sensitizers using a panel of methacrylates.

    NARCIS (Netherlands)

    Rustemeyer, T.; Preuss, M; Blomberg - van der Flier, von B.M.E.; Das, PK; Scheper, R.J.

    2003-01-01

    Allergen-induced emigration and maturation of dendritic cells (DC) are pivotal steps in sparking off allergic contact dermatitis. In vitro models, reflecting these steps, may provide tools for assessment of sensitizing capacities of putative contact allergens. Here, we evaluated the applicability of

  14. Cascades in helical turbulence

    CERN Document Server

    Ditlevsen, P D

    2001-01-01

    The existence of a second quadratic inviscid invariant, the helicity, in a turbulent flow leads to coexisting cascades of energy and helicity. An equivalent of the four-fifth law for the longitudinal third order structure function, which is derived from energy conservation, is easily derived from helicity conservation cite{Procaccia,russian}. The ratio of dissipation of helicity to dissipation of energy is proportional to the wave-number leading to a different Kolmogorov scale for helicity than for energy. The Kolmogorov scale for helicity is always larger than the Kolmogorov scale for energy so in the high Reynolds number limit the flow will always be helicity free in the small scales, much in the same way as the flow will be isotropic and homogeneous in the small scales. A consequence is that a pure helicity cascade is not possible. The idea is illustrated in a shell model of turbulence.

  15. Evaluation of human embryonic stem cells and their differentiated fibroblastic progenies as cellular models for in vitro genotoxicity screening.

    Science.gov (United States)

    Vinoth, Kumar Jayaseelan; Manikandan, Jayapal; Sethu, Swaminathan; Balakrishnan, Lakshmidevi; Heng, Alexis; Lu, Kai; Hande, Manoor Prakash; Cao, Tong

    2014-08-20

    This study evaluated human embryonic stem cells (hESC) and their differentiated fibroblastic progenies as cellular models for genotoxicity screening. The DNA damage response of hESCs and their differentiated fibroblastic progenies were compared to a fibroblastic cell line (HEPM, CRL1486) and primary cultures of peripheral blood lymphocytes (PBL), upon exposure to Mitomycin C, gamma irradiation and H2O2. It was demonstrated that hESC-derived fibroblastic progenies (H1F) displayed significantly higher chromosomal aberrations, micronuclei formation and double strand break (DSB) formation, as compared to undifferentiated hESC upon exposure to genotoxic stress. Nevertheless, H1F cell types displayed comparable sensitivities to genotoxic challenge as HEPM and PBL, both of which are representative of somatic cell types commonly used for genotoxicity screening. Subsequently, transcriptomic and pathways analysis identified differential expression of critical genes involved in cell death and DNA damage response upon exposure to gamma irradiation. The results thus demonstrate that hESC-derived fibroblastic progenies are as sensitive as commonly-used somatic cell types for genotoxicity screening. Moreover, hESCs have additional advantages, such as their genetic normality compared to immortalized cell lines, as well as their amenability to scale-up for producing large, standardized quantities of cells for genotoxicity screening on an industrial scale, something which can never be achieved with primary cell cultures. Copyright © 2014. Published by Elsevier B.V.

  16. Psi-screen, an in vitro toxicity test system: applications in the bioassay of perfumes and fragrance chemicals.

    Science.gov (United States)

    Griffiths, David E

    2005-10-01

    The effects of 65 perfume formulations (perfume oils, perfumes, eau de parfum, eau de toilette) on mitochondrial membrane potential (Psim) and mitochondrial respiration have been investigated using a mitochondria-based assay for (Psim, termed Psi-Screen. All the perfume formulations tested are highly active in the Psi-Screen assay, and the major site of inhibition in all cases is NADH-ubiquinone reductase (Complex I). This is confirmed in studies on the inhibition of NADH oxidase and NADH-ubiquinone reductase. Some formulations also inhibit succinate oxidation at either Complex II or Complex III. Evidence for the inhibition of mitochondrial ATPase is presented, as well as for the induction of reactive oxygen species production by perfume inhibition of Complex I. Thus, perfume formulations are multiple inhibitor mixtures which inhibit multiple bioenergetic functions at high dilutions. The implications of these findings are discussed with respect to cell toxicity via necrosis and/or apoptosis. Twenty candidate fragrance chemicals were investigated and all inhibited Complex I (5 at <35 microM). Mass screening strategies and high-throughput screening assays are discussed.

  17. Unique Nanoparticle Optical Properties Confound Fluorescent Based Assays Widely Employed in Their In Vitro Toxicity Screening and Ranking

    Science.gov (United States)

    Nanoparticles (NPs) are novel materials having at least one dimension less than 100 nm and display unique physicochemical properties due to their nanoscale size. An emphasis has been placed on developing high throughput screening (HTS) assays to characterize and rank the toxiciti...

  18. Screening the banana biodiversity for drought tolerance: can an in vitro growth model and proteomics be used as a tool to discover tolerant varieties and understand homeostasis

    Directory of Open Access Journals (Sweden)

    Anne-Catherine eVanhove

    2012-08-01

    Full Text Available There is a great need for research aimed at understanding drought tolerance, screening for drought tolerant varieties and breeding cops with an improved water use efficiency. Bananas and plantains are a major staple food and export product with a worldwide production of over 135 million tonnes per year. Water however is the most limiting abiotic factor in banana production. A screening of the Musa biodiversity has not yet been performed. We at KU Leuven host the Musa International Germplasm collection with over 1200 accessions. To screen the Musa biodiversity for drought tolerant varieties, we developed a screening test for in vitro plants. Five varieties representing different genomic constitutions in banana (AAAh, AAA, AAB, AABp and ABB were selected and subjected to a mild osmotic stress. The ABB variety showed the smallest stress induced growth reduction. To get an insight into the acclimation and the accomplishment of homeostasis, the leaf proteome of this variety was characterized via 2D DIGE. After extraction of the leaf proteome of 6 control and 6 stressed plants, 2600 spots could be distinguished. A PCA analysis indicates that control and stressed plants can blindly be classified based on their proteome. One hundred and twelve proteins were significantly more abundant in the stressed plants and eighteen proteins were significantly more abundant in control plants (FDR α 0.05. Twenty four differential proteins could be identified. The proteome analysis clearly shows that there is a new balance in the stressed plants and that the respiration, metabolism of ROS and several dehydrogenases involved in NAD/NADH homeostasis play an important role.

  19. Screening the banana biodiversity for drought tolerance: can an in vitro growth model and proteomics be used as a tool to discover tolerant varieties and understand homeostasis.

    Science.gov (United States)

    Vanhove, Anne-Catherine; Vermaelen, Wesley; Panis, Bart; Swennen, Rony; Carpentier, Sebastien C

    2012-01-01

    There is a great need for research aimed at understanding drought tolerance, screening for drought tolerant varieties and breeding crops with an improved water use efficiency. Bananas and plantains are a major staple food and export product with a worldwide production of over 135 million tonnes per year. Water however is the most limiting abiotic factor in banana production. A screening of the Musa biodiversity has not yet been performed. We at KU Leuven host the Musa International Germplasm collection with over 1200 accessions. To screen the Musa biodiversity for drought tolerant varieties, we developed a screening test for in vitro plants. Five varieties representing different genomic constitutions in banana (AAAh, AAA, AAB, AABp, and ABB) were selected and subjected to a mild osmotic stress. The ABB variety showed the smallest stress induced growth reduction. To get an insight into the acclimation and the accomplishment of homeostasis, the leaf proteome of this variety was characterized via 2D DIGE. After extraction of the leaf proteome of six control and six stressed plants, 2600 spots could be distinguished. A PCA analysis indicates that control and stressed plants can blindly be classified based on their proteome. One hundred and twelve proteins were significantly more abundant in the stressed plants and 18 proteins were significantly more abundant in control plants (FDR α 0.05). Twenty four differential proteins could be identified. The proteome analysis clearly shows that there is a new balance in the stressed plants and that the respiration, metabolism of ROS and several dehydrogenases involved in NAD/NADH homeostasis play an important role.

  20. Screening of traditional antidiabetic medicinal plants of Mauritius for possible alpha-amylase inhibitory effects in vitro.

    Science.gov (United States)

    Kotowaroo, M I; Mahomoodally, M F; Gurib-Fakim, A; Subratty, A H

    2006-03-01

    In this study, seven exotic/indigenous medicinal plants of Mauritius, namely Coix lacryma-jobi (Poaceae), Aegle marmelos (Rutaceae), Artocarpus heterophyllus (Moraceae), Vangueria madagascariensis (Rubiaceae), Azadirachta indica (Meliaceae), Eriobotrya japonica (Rosaceae) and Syzigium cumini (Myrtaceae) were studied for possible effects on starch breakdown by alpha-amylase in vitro. The results showed that only Artocarpus heterophyllus significantly (p Artocarpus heterophyllus on alpha-amylase activity using rat plasma in vitro. It was found that the aqueous leaf extract significantly (p Artocarpus heterophyllus behaved as a competitive inhibitor. Results from the present study tend to indicate that Artocarpus heterophyllus could act as a 'starch blocker' thereby reducing post-prandial glucose peaks. Copyright 2006 John Wiley & Sons, Ltd.

  1. Comparison of Two Static in Vitro Digestion Methods for Screening the Bioaccessibility of Carotenoids in Fruits, Vegetables, and Animal Products.

    Science.gov (United States)

    Rodrigues, Daniele B; Chitchumroonchokchai, Chureeporn; Mariutti, Lilian R B; Mercadante, Adriana Z; Failla, Mark L

    2017-12-27

    In vitro digestion methods are routinely used to assess the bioaccessibility of carotenoids and other dietary lipophilic compounds. Here, we compared the recovery of carotenoids and their efficiency of micellarization in digested fruits, vegetables, egg yolk, and salmon and also in mixed-vegetable salads with and without either egg yolk or salmon using the static INFOGEST method22 and the procedure of Failla et al.16 Carotenoid stability during the simulated digestion was ≥70%. The efficiencies of the partitioning of carotenoids into mixed micelles were similar when individual plant foods and salad meals were digested using the two static methods. Furthermore, the addition of cooked egg or salmon to vegetable salads increased the bioaccessibility of some carotenoids. Our findings showed that the two methods of in vitro digestion generated similar estimates of carotenoid retention and bioaccessibility for diverse foods.

  2. Estimation of naphthaquinones from Arnebia hispidissima (Lehm.) DC. In vivo and in vitro. I. Anti-inflammatory screening.

    Science.gov (United States)

    Singh, B; Sahu, P M; Jain, S C; Singh, S

    2004-02-01

    The hexane extract of Arnebia hispidissima yielded a mixture of naphthaquinones: arnebin-1, arnebin-7, tiglic acid (ester of dihydroxy alkannin), alkannin, arnebinol and cycloarnebin-7. To evaluate the anti-inflammatory activities of hexane extract and isolated naphthaquinones, models with carrageenan-induced acute arthritis and complete Freund's adjuvant (CFA)-induced chronic arthritis in rats were conducted. The observed results indicated that pretreatment with cycloarnebin-7 significantly inhibited the carrageenan-induced acute arthritis. Moreover, arnebin-1, significantly suppressed the development of chronic arthritis induced by CFA. The present study deals with the quantification of naphthaquinones from in vivo and in vitro cell cultures of plant species and isolated compounds from intact plant tested for their swelling inhibitory potency. It has been reported that arnebin-1 was the major naphthaquinone both in vivo (0.62%) and in vitro (0.27%) cell cultures. Copyright 2004 John Wiley & Sons, Ltd.

  3. In vitro acute and developmental neurotoxicity screening: an overview of cellular platforms and high-throughput technical possibilities.

    Science.gov (United States)

    Schmidt, Béla Z; Lehmann, Martin; Gutbier, Simon; Nembo, Erastus; Noel, Sabrina; Smirnova, Lena; Forsby, Anna; Hescheler, Jürgen; Avci, Hasan X; Hartung, Thomas; Leist, Marcel; Kobolák, Julianna; Dinnyés, András

    2017-01-01

    Neurotoxicity and developmental neurotoxicity are important issues of chemical hazard assessment. Since the interpretation of animal data and their extrapolation to man is challenging, and the amount of substances with information gaps exceeds present animal testing capacities, there is a big demand for in vitro tests to provide initial information and to prioritize for further evaluation. During the last decade, many in vitro tests emerged. These are based on animal cells, human tumour cell lines, primary cells, immortalized cell lines, embryonic stem cells, or induced pluripotent stem cells. They differ in their read-outs and range from simple viability assays to complex functional endpoints such as neural crest cell migration. Monitoring of toxicological effects on differentiation often requires multiomics approaches, while the acute disturbance of neuronal functions may be analysed by assessing electrophysiological features. Extrapolation from in vitro data to humans requires a deep understanding of the test system biology, of the endpoints used, and of the applicability domains of the tests. Moreover, it is important that these be combined in the right way to assess toxicity. Therefore, knowledge on the advantages and disadvantages of all cellular platforms, endpoints, and analytical methods is essential when establishing in vitro test systems for different aspects of neurotoxicity. The elements of a test, and their evaluation, are discussed here in the context of comprehensive prediction of potential hazardous effects of a compound. We summarize the main cellular characteristics underlying neurotoxicity, present an overview of cellular platforms and read-out combinations assessing distinct parts of acute and developmental neurotoxicology, and highlight especially the use of stem cell-based test systems to close gaps in the available battery of tests.

  4. Identification of a new class of FtsZ inhibitors by structure-based design and in vitro screening.

    Science.gov (United States)

    Chan, Fung-Yi; Sun, Ning; Neves, Marco A C; Lam, Polo Chun-Hung; Chung, Wai-Hong; Wong, Lai-King; Chow, Ho-Yin; Ma, Dik-Lung; Chan, Pak-Ho; Leung, Yun-Chung; Chan, Tak-Hang; Abagyan, Ruben; Wong, Kwok-Yin

    2013-08-26

    The Filamenting temperature-sensitive mutant Z (FtsZ), an essential GTPase in bacterial cell division, is highly conserved among Gram-positive and Gram-negative bacteria and thus considered an attractive target to treat antibiotic-resistant bacterial infections. In this study, a new class of FtsZ inhibitors bearing the pyrimidine-quinuclidine scaffold was identified from structure-based virtual screening of natural product libraries. Iterative rounds of in silico studies and biological evaluation established the preliminary structure-activity relationships of the new compounds. Potent FtsZ inhibitors with low micromolar IC₅₀ and antibacterial activity against S. aureus and E. coli were found. These findings support the use of virtual screening and structure-based design for the rational development of new antibacterial agents with innovative mechanisms of action.

  5. In vitro screening of inhibition of PPAR-γ activity as a first step in identification of potential breast carcinogens

    DEFF Research Database (Denmark)

    Kopp, Tine Iskov; Lundqvist, J.; Petersen, R. K.

    2015-01-01

    measured in the H295R steroidogenesis assay after incubation with the chemicals. Ethylene glycol, ethyl acetate, and dimethyl sulphoxide inhibited PPAR-γ transactivation in a dose-dependent manner. The inhibitory effect on PPAR-γ was specific for PPAR-γ since the AB domain of PPAR-γ was required...... for the inhibitory effect. In the second step, ethylene glycol significantly increased production of oestradiol by 19% (p ethyl acetate inhibited production of testosterone (p ... followed by a well-established steroidogenesis assay for production of sex hormones in exposed H295 R cells may provide a screening tool for potential breast carcinogens. This initial screening thus identified ethylene glycol and possibly ethyl acetate as potential breast carcinogens....

  6. The establishment of in?vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum

    OpenAIRE

    Suganuma, Keisuke; Yamasaki, Shino; Molefe, Nthatisi Innocentia; Musinguzi, Peter Simon; Davaasuren, Batdorj; Mossaad, Ehab; Narantsatsral, Sandagdorj; Battur, Banzragch; Battsetseg, Badgar; Inoue, Noboru

    2017-01-01

    Dourine is caused by Trypanosoma equiperdum via coitus with an infected horse. Although dourine is distributed in Equidae worldwide and is listed as an internationally important animal disease by the World Organization for Animal Health (OIE), no effective treatment strategies have been established. In addition, there are no reports on drug discovery, because no drug screening system exists for this parasite. A new T.?equiperdum strain was recently isolated from the genital organ of a stallio...

  7. Identification and quantification of estrogenic compounds in recycled and virgin paper for household use as determined by an in vitro yeast estrogen screen and chemical analysis

    DEFF Research Database (Denmark)

    Vinggaard, Anne; Körner, Wolfgang; Lund, Kirsten H.

    2000-01-01

    The use of recycled paper for the manufacture of food contact materials is widespread, but very little is known about the presence of potential contaminants in the paper. The purpose of this study was to assess the worst-case migration of estrogenic active compounds using extracts of paper...... contain bisphenol A and other xenoestrogens may apply to other types of recycled paper used for food packaging and emphasize the importance of identifying this and other contaminants in recycled paper in general. These data indicate that bisphenol A may be useful as a purity indicator for recycled paper....... for household use. Twenty different brands of kitchen rolls, nine of which were made from recycled paper and the remainder from virgin paper, were obtained from retail shops. Paper extracts were subjected to (a) determination of the total estrogenic activity by using an in vitro estrogen screen based on yeast...

  8. Peptides Derived from Soy and Lupin Protein as Dipeptidyl-Peptidase IV Inhibitors: In Vitro Biochemical Screening and in Silico Molecular Modeling Study.

    Science.gov (United States)

    Lammi, Carmen; Zanoni, Chiara; Arnoldi, Anna; Vistoli, Giulio

    2016-12-28

    Dipeptidyl peptidase IV (DPP-IV) is a new molecular target correlated with the development of type 2 diabetes. Literature describes the identification of some inhibitory peptides from the hydrolysis of different food proteins. This article reports a study on six peptides from soybean and lupin proteins, i.e., Soy 1 (IAVPTGVA), Soy 2 (YVVNPDNDEN), Soy 3 (YVVNPDNNEN), Lup 1 (LTFPGSAED), Lup 2 (LILPKHSDAD), and Lup 3 (GQEQSHQDEGVIVR), which were screened for their capacity to inhibit the activity of DPP-IV, using an in vitro bioassay against human recombinant DPP-IV. Two peptides Soy 1 and Lup 1 resulted to be efficient inhibitors with IC50 values equal to 106 and 228 μM, respectively. A molecular docking analysis predicted the key molecular interactions, stabilizing the active peptides within DPP-IV enzyme. Soy and lupin proteins may be sources of DPP-IV inhibitory peptides potentially useful for the prevention of type 2 diabetes.

  9. Quartz-Containing Ceramic Dusts: In vitro screening of the cytotoxic, genotoxic and pro-inflammatory potential of 5 factory samples

    Science.gov (United States)

    Ziemann, C.; Jackson, P.; Brown, R.; Attik, G.; Rihn, B. H.; Creutzenberg, O.

    2009-02-01

    Inhalation of some respirable crystalline silica (MMAD RF>TG>Ti>BR>TC>Al2O3. DNA-damage was maximal for BR and TI followed by DQ12>TG>TC>RF>Al2O3. All dusts induced PGE2-liberation (DQ12>BR>TC>TG>Ti>RF>Al2O3) at 50μg/cm2 (4h), but TNF-a mRNA (10μg/cm2, 24h) was only increased by DQ12, TG (quartz-dependently), and TC. In conclusion, these in vitro tests were an adequate approach to screen the toxic potential of quartz-containing ceramic dusts, but the quartz-content was too low to differentiate the various quartz-varieties.

  10. A novel inhibitor of human La protein with anti-HBV activity discovered by structure-based virtual screening and in vitro evaluation.

    Directory of Open Access Journals (Sweden)

    Jing Tang

    Full Text Available BACKGROUND: Over 350 million people worldwide are infected with hepatitis B virus (HBV, a major cause of liver failure and hepatocellular carcinoma. Current therapeutic agents are highly effective, but are also associated with development of viral resistance. Therefore, strategies for identifying other anti-HBV agents with specific, but distinctive mechanisms of action are needed. The human La (hLa protein, which forms a stabilizing complex with HBV RNA ribonucleoprotein to promote HBV replication, is a promising target of molecular therapy. AIMS: This study aimed to discover novel inhibitors of hLa that could inhibit HBV replication and expression. METHODS: A multistage molecular docking approach was used to screen a Specs database and an in-house library against hLa binding sites. Sequential in vitro evaluations were performed to detect potential compounds with high scores in HepG2.2.15 cells. RESULTS: Of the 26 potential compounds with high scores chosen for experimental verification, 12 had HBV DNA inhibition ratios of less than 50% with P<0.05. Six had significant inhibition of HBV e antigen (HBeAg levels, and 13 had significant inhibition of HBV surface antigen (HBsAg levels by in vitro assays. Compounds HBSC-11, HBSC-15 and HBSC-34 (HBSC is system prefix for active compounds screened by the library were selected for evaluation. HBSC-11 was found to have an obvious inhibitory effect on hLa transcription and expression. CONCLUSIONS: Our findings suggest that anti-HBV activity of HBSC-11 may be mediated by a reduction in hLa levels. In addition, our data suggest the potential clinical use of hLa inhibitors, such as HBSC-11, for treating HBV infection.

  11. A Drug Combination Screen Identifies Drugs Active against Amoxicillin-Induced Round Bodies of In Vitro Borrelia burgdorferi Persisters from an FDA Drug Library.

    Science.gov (United States)

    Feng, Jie; Shi, Wanliang; Zhang, Shuo; Sullivan, David; Auwaerter, Paul G; Zhang, Ying

    2016-01-01

    Although currently recommended antibiotics for Lyme disease such as doxycycline or amoxicillin cure the majority of the patients, about 10-20% of patients treated for Lyme disease may experience lingering symptoms including fatigue, pain, or joint and muscle aches. Under experimental stress conditions such as starvation or antibiotic exposure, Borrelia burgdorferi can develop round body forms, which are a type of persister bacteria that appear resistant in vitro to customary first-line antibiotics for Lyme disease. To identify more effective drugs with activity against the round body form of B. burgdorferi, we established a round body persister model induced by exposure to amoxicillin (50 μg/ml) and then screened the Food and Drug Administration drug library consisting of 1581 drug compounds and also 22 drug combinations using the SYBR Green I/propidium iodide viability assay. We identified 23 drug candidates that have higher activity against the round bodies of B. burgdorferi than either amoxicillin or doxycycline. Eleven individual drugs scored better than metronidazole and tinidazole which have been previously described to be active against round bodies. In this amoxicillin-induced round body model, some drug candidates such as daptomycin and clofazimine also displayed enhanced activity which was similar to a previous screen against stationary phase B. burgdorferi persisters not exposure to amoxicillin. Additional candidate drugs active against round bodies identified include artemisinin, ciprofloxacin, nifuroxime, fosfomycin, chlortetracycline, sulfacetamide, sulfamethoxypyridazine and sulfathiozole. Two triple drug combinations had the highest activity against amoxicillin-induced round bodies and stationary phase B. burgdorferi persisters: artemisinin/cefoperazone/doxycycline and sulfachlorpyridazine/daptomycin/doxycycline. These findings confirm and extend previous findings that certain drug combinations have superior activity against B. burgdorferi

  12. Prevalence, specificity and determinants of lipid-interacting PDZ domains from an in-cell screen and in vitro binding experiments.

    Directory of Open Access Journals (Sweden)

    Ylva Ivarsson

    Full Text Available BACKGROUND: PDZ domains are highly abundant protein-protein interaction modules involved in the wiring of protein networks. Emerging evidence indicates that some PDZ domains also interact with phosphoinositides (PtdInsPs, important regulators of cell polarization and signaling. Yet our knowledge on the prevalence, specificity, affinity, and molecular determinants of PDZ-PtdInsPs interactions and on their impact on PDZ-protein interactions is very limited. METHODOLOGY/PRINCIPAL FINDINGS: We screened the human proteome for PtdInsPs interacting PDZ domains by a combination of in vivo cell-localization studies and in vitro dot blot and Surface Plasmon Resonance (SPR experiments using synthetic lipids and recombinant proteins. We found that PtdInsPs interactions contribute to the cellular distribution of some PDZ domains, intriguingly also in nuclear organelles, and that a significant subgroup of PDZ domains interacts with PtdInsPs with affinities in the low-to-mid micromolar range. In vitro specificity for the head group is low, but with a trend of higher affinities for more phosphorylated PtdInsPs species. Other membrane lipids can assist PtdInsPs-interactions. PtdInsPs-interacting PDZ domains have generally high pI values and contain characteristic clusters of basic residues, hallmarks that may be used to predict additional PtdInsPs interacting PDZ domains. In tripartite binding experiments we established that peptide binding can either compete or cooperate with PtdInsPs binding depending on the combination of ligands. CONCLUSIONS/SIGNIFICANCE: Our screen substantially expands the set of PtdInsPs interacting PDZ domains, and shows that a full understanding of the biology of PDZ proteins will require a comprehensive insight into the intricate relationships between PDZ domains and their peptide and lipid ligands.

  13. LCMS and GCMS for the screening of alkaloids in natural and in vitro extracts of Leucojum aestivum.

    Science.gov (United States)

    Ptak, Agata; El Tahchy, Anna; Dupire, François; Boisbrun, Michel; Henry, Max; Chapleur, Yves; Moś, Maria; Laurain-Mattar, Dominique

    2009-01-01

    HPLC coupled to a mass spectrometer (MS) was used for the analysis of galanthamine and lycorine in natural extracts of Leucojum aestivum and in their in vitro cultures grown with a precursor (ACC), inhibitors (AgNO(3), STS), or an absorber (KMnO(4)) of ethylene. The maximum galanthamine (0.002%) and lycorine (0.02%) concentrations in tissue cultures were obtained in the presence of KMnO(4). GCMS was used to investigate underivatized alkaloid mixtures from L. aestivum. Seven alkaloids were identified in in vivo bulbs. KMnO(4) led to the highest diversity of alkaloids in tissue culture extracts.

  14. The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum.

    Science.gov (United States)

    Suganuma, Keisuke; Yamasaki, Shino; Molefe, Nthatisi Innocentia; Musinguzi, Peter Simon; Davaasuren, Batdorj; Mossaad, Ehab; Narantsatsral, Sandagdorj; Battur, Banzragch; Battsetseg, Badgar; Inoue, Noboru

    2017-08-01

    Dourine is caused by Trypanosoma equiperdum via coitus with an infected horse. Although dourine is distributed in Equidae worldwide and is listed as an internationally important animal disease by the World Organization for Animal Health (OIE), no effective treatment strategies have been established. In addition, there are no reports on drug discovery, because no drug screening system exists for this parasite. A new T. equiperdum strain was recently isolated from the genital organ of a stallion that showed typical symptoms of dourine. In the present study, we adapted T. equiperdum IVM-t1 from soft agarose media to HMI-9 liquid media to develop a drug screening assay for T. equiperdum. An intracellular ATP-based luciferase assay using CellTiter-Glo reagent and an intracellular dehydrogenase activity-based colorimetric assay using WTS-8 tetrazolium salt (CCK-8 reagent) were used in order to examine the trypanocidal effects of each compound. In addition, the IC 50 values of 4 reference trypanocidal compounds (pentamidine, diminazene, suramin and melarsomine) were evaluated and compared using established assays. The IC 50 values of these reference compounds corresponded well to previous studies involving other strains of T. equiperdum. The luciferase assay would be suitable for the mass screening of chemical libraries against T. equiperdum because it allows for the simple and rapid-evaluation of the trypanocidal activities of test compounds, while a simple, inexpensive colorimetric assay will be applicable in developing countries for the evaluation of the drug sensitivity of epidemic trypanosome strains. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  15. The establishment of in vitro culture and drug screening systems for a newly isolated strain of Trypanosoma equiperdum

    Directory of Open Access Journals (Sweden)

    Keisuke Suganuma

    2017-08-01

    Full Text Available Dourine is caused by Trypanosoma equiperdum via coitus with an infected horse. Although dourine is distributed in Equidae worldwide and is listed as an internationally important animal disease by the World Organization for Animal Health (OIE, no effective treatment strategies have been established. In addition, there are no reports on drug discovery, because no drug screening system exists for this parasite. A new T. equiperdum strain was recently isolated from the genital organ of a stallion that showed typical symptoms of dourine. In the present study, we adapted T. equiperdum IVM-t1 from soft agarose media to HMI-9 liquid media to develop a drug screening assay for T. equiperdum. An intracellular ATP-based luciferase assay using CellTiter-Glo reagent and an intracellular dehydrogenase activity-based colorimetric assay using WTS-8 tetrazolium salt (CCK-8 reagent were used in order to examine the trypanocidal effects of each compound. In addition, the IC50 values of 4 reference trypanocidal compounds (pentamidine, diminazene, suramin and melarsomine were evaluated and compared using established assays. The IC50 values of these reference compounds corresponded well to previous studies involving other strains of T. equiperdum. The luciferase assay would be suitable for the mass screening of chemical libraries against T. equiperdum because it allows for the simple and rapid-evaluation of the trypanocidal activities of test compounds, while a simple, inexpensive colorimetric assay will be applicable in developing countries for the evaluation of the drug sensitivity of epidemic trypanosome strains.

  16. Fractal Levy correlation cascades

    Energy Technology Data Exchange (ETDEWEB)

    Eliazar, Iddo [Department of Technology Management, Holon Institute of Technology, Holon 58102 (Israel); Klafter, Joseph [School of Chemistry, Sackler Faculty of Exact Sciences, Tel Aviv University, Tel Aviv 69978 (Israel)

    2007-04-20

    The correlation structure of a wide class of random processes, driven by stable non-Gaussian Levy noise sources, is explored. Since these noises are of infinite variance, correlations cannot be measured via auto-covariance functions. Exploiting the underlying Poissonian structure of Levy noises, we present a cascade of 'Poissonian correlation functions' which characterize the correlation structure and the process distribution of the processes under consideration. The theory developed is applied to various examples including motions, Ornstein-Uhlenbeck and moving-average processes, and fractional motions and noises-all driven by stable non-Gaussian Levy noises. (fast track communication)

  17. Cascading Corruption News

    DEFF Research Database (Denmark)

    Damgaard, Mads

    2018-01-01

    Through a content analysis of 8,800 news items and six months of front pages in three Brazilian newspapers, all dealing with corruption and political transgression, this article documents the remarkable skew of media attention to corruption scandals. The bias is examined as an information...... phenomenon, arising from systemic and commercial factors of Brazil’s news media: An information cascade of news on corruption formed, destabilizing the governing coalition and legitimizing the impeachment process of Dilma Rousseff. As this process gained momentum, questions of accountability were disregarded...

  18. Canine toys and training devices as sources of exposure to phthalates and bisphenol A: quantitation of chemicals in leachate and in vitro screening for endocrine activity.

    Science.gov (United States)

    Wooten, Kimberly J; Smith, Philip N

    2013-11-01

    Chewing and mouthing behaviors exhibited by pet dogs are likely to lead to oral exposures to a variety of environmental chemicals. Products intended for chewing and mouthing uses include toys and training devices that are often made of plastics. The goal of the current study was to determine if a subset of phthalates and bisphenol A (BPA), endocrine disrupting chemicals commonly found in plastics, leach out of dog toys and training devices (bumpers) into synthetic canine saliva. In vitro assays were used to screen leachates for endocrine activity. Bumper leachates were dominated by di-2-ethylhexyl phthalate (DEHP) and BPA, with concentrations reaching low μg mL(-1) following short immersions in synthetic saliva. Simulated chewing of bumpers during immersion in synthetic saliva increased concentrations of phthalates and BPA as compared to new bumpers, while outdoor storage had variable effects on concentrations (increased DEHP; decreased BPA). Toys leached substantially lower concentrations of phthalates and BPA, with the exception of one toy which leached considerable amounts of diethyl phthalate. In vitro assays indicated anti-androgenic activity of bumper leachates, and estrogenic activity of both bumper and toy leachates. These results confirm that toys and training devices are potential sources of exposure to endocrine disrupting chemicals in pet dogs. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Probiotic attributes of indigenous Lactobacillus spp. isolated from traditional fermented foods and beverages of north-western Himalayas using in vitro screening and principal component analysis.

    Science.gov (United States)

    Kumari, Anila; Angmo, Kunzes; Monika; Bhalla, Tek Chand

    2016-05-01

    The present research was designed to explore indigenous probiotic Lactic acid bacteria from traditional fermented foods and beverages of North-western Himalayas for their probiotic potential. It was achieved through a step-by step approach focused on the technological characterization, evaluation of the probiotic traits and adherence ability. Fifty one LAB isolates from traditional fermented foods and beverages were initially screened for their technological properties and among them twenty isolates were selected. These isolates were further characterized and identified using 16S rRNA gene sequencing as Lactobacillus brevis (7 isolates), Lactobacillus casei (5), Lactobacillus paracasei (2), Lactobacillus buchneri (1), Lactobacillus plantarum (1) and Lactobacillus sp. (3). Identified isolates were evaluated by in vitro methods including survival in gastrointestinal tract, antibiotic susceptibility, antimicrobial activity, cell surface characteristics, exopolysacharride production and haemolytic activity. The results of these experiments were used as input data for Principal Component Analysis; thus, to select the most promising probiotic isolates. Three isolates (L. brevis PLA2, L. paracasei PLA8 and L. brevis PLA16) were found to be most technological relevant and promising probiotic candidates in comparison to commercial probiotic strains. L. brevis PLA2 was selected as best isolate with probiotic potential by in vitro adherence to the human intestinal HT-29 cell line.

  20. In vitro screening of selected probiotic properties of Lactobacillus strains isolated from traditional fermented cabbage and cucumber.

    Science.gov (United States)

    Zielińska, Dorota; Rzepkowska, Anna; Radawska, Anna; Zieliński, Konrad

    2015-02-01

    Most important during probiotic selection are gastric acid and bile tolerance, the adhesion to the luminal epithelium to colonize the lower gastrointestinal tract of a human and safety for human consumption. The aim of this study was to evaluate the selected probiotic in vitro properties of Lactobacillus spp. Strains isolated from traditional fermented food. A total 38 strains were isolated from the pickled samples and 14 were identified as Lactobacillus spp. The survival of almost all strains after incubation at pH 2.5 did not change markedly, and remained at above 90 % (10(9) CFU/mL). The strains also exhibited a high survival rate at pH 3.5 (>90 %), whereas pH 1.5 all were died. Just four strains could survive 90 min. at pH 1.5 (Safety evaluation excluded four of tested strains as candidate probiotics, according to antibiotic resistance patterns and certain metabolic activities. On the basis on the results 10 of the selected Lactobacillus strains are safe and can survive under gastrointestinal conditions, which requires them to future in vitro and in vivo probiotic studies.

  1. An in vitro transport model for rapid screening and predicting the permeability of candidate compounds at blood-brain barrier.

    Science.gov (United States)

    Yang, Zhi-Hong; Sun, Xiao; Mei, Chao; Sun, Xiao-Bo; Liu, Xiao-Dong; Chang, Qi

    2011-12-01

    The aim of this study was to design and develop a simple in vitro blood-brain barrier (BBB) permeation model for elementarily and rapidly predicting the permeability of candidate compounds at BBB and further evaluating whether P-glycoprotein (P-gp) affects them across BBB. The model was mainly composed of cultured rat brain microvascular endothelial cells (rBMECs), glass contraption, and micropore membrane. First, we evaluated the model by morphological observation. Second, the restriction effects of paracellular transport were verified by measuring marker probes transport, and monitoring transendothelial electrical resistance (TEER) and leakage. Finally, protein expression and activity of P-gp were confirmed by carrying out Western blot analysis and polarized transport of rhodamine-123 (Rho123) in rBMECs. The rBMECs retained both endothelial cells and BBB features. The rBMECs model reproducibly attained approximately 130 Ω cm² on the steady-state TEER value, and displayed a barrier function to marker probes transport by decreasing the permeability. Protein band of 170 kDa manifested the existence of P-gp in the rBMECs, and the findings of cyclosporin A-sensitive decrease of Rho123 efflux confirmed the presence of P-gp activity. A simple, rapid, and convenient in vitro BBB permeation model was successfully established and applied to evaluate the BBB transport profiles of three natural flavonoids: quercetin, naringenin, and rutin.

  2. Screening of toxic potential of graphene family nanomaterials using in vitro and alternative in vivo toxicity testing systems.

    Science.gov (United States)

    Chatterjee, Nivedita; Yang, Ji Su; Park, Kwangsik; Oh, Seung Min; Park, Jeonggue; Choi, Jinhee

    2015-01-01

    The widely promising applications of graphene nanomaterials raise considerable concerns regarding their environmental and human health risk assessment. The aim of the current study was to evaluate the toxicity profiling of graphene family nananomaterials (GFNs) in alternative in vitro and in vivo toxicity testing models. The GFNs used in this study are graphene nanoplatelets ([GNPs]-pristine, carboxylate [COOH] and amide [NH2]) and graphene oxides (single layer [SLGO] and few layers [FLGO]). The human bronchial epithelial cells (Beas2B cells) as in vitro system and the nematode Caenorhabditis elegans as in vivo system were used to profile the toxicity response of GFNs. Cytotoxicity assays, colony formation assay for cellular toxicity and reproduction potentiality in C. elegans were used as end points to evaluate the GFNs' toxicity. In general, GNPs exhibited higher toxicity than GOs in Beas2B cells, and among the GNPs the order of toxicity was pristine>NH2>COOH. Although the order of toxicity of the GNPs was maintained in C. elegans reproductive toxicity, but GOs were found to be more toxic in the worms than GNPs. In both systems, SLGO exhibited profoundly greater dose dependency than FLGO. The possible reason of their differential toxicity lay in their distinctive physicochemical characteristics and agglomeration behavior in the exposure media. The present study revealed that the toxicity of GFNs is dependent on the graphene nanomaterial's physical forms, surface functionalizations, number of layers, dose, time of exposure and obviously, on the alternative model systems used for toxicity assessment.

  3. Putative histidine kinase inhibitors with antibacterial effect against multi-drug resistant clinical isolates identified by in vitro and in silico screens

    Science.gov (United States)

    Velikova, Nadya; Fulle, Simone; Manso, Ana Sousa; Mechkarska, Milena; Finn, Paul; Conlon, J. Michael; Oggioni, Marco Rinaldo; Wells, Jerry M.; Marina, Alberto

    2016-05-01

    Novel antibacterials are urgently needed to address the growing problem of bacterial resistance to conventional antibiotics. Two-component systems (TCS) are widely used by bacteria to regulate gene expression in response to various environmental stimuli and physiological stress and have been previously proposed as promising antibacterial targets. TCS consist of a sensor histidine kinase (HK) and an effector response regulator. The HK component contains a highly conserved ATP-binding site that is considered to be a promising target for broad-spectrum antibacterial drugs. Here, we describe the identification of putative HK autophosphorylation inhibitors following two independent experimental approaches: in vitro fragment-based screen via differential scanning fluorimetry and in silico structure-based screening, each followed up by the exploration of analogue compounds as identified by ligand-based similarity searches. Nine of the tested compounds showed antibacterial effect against multi-drug resistant clinical isolates of bacterial pathogens and include three novel scaffolds, which have not been explored so far in other antibacterial compounds. Overall, putative HK autophosphorylation inhibitors were found that together provide a promising starting point for further optimization as antibacterials.

  4. Phytochemical and in vitro screening of some Ficus and Morus spp. for hypolipidaemic and antioxidant activities and in vivo assessment of Ficus mysorensis (Roth).

    Science.gov (United States)

    Awad, Nagwa E; Seida, Ahmed A; Hamed, Manal A; Mahmoud, Ahlam H; Elbatanony, Marwa M

    2012-01-01

    Phytochemical screening of air-dried leaves and fruit juice of certain Ficus and Morus spp. have been studied. In an in vitro study, the ethanol and hexane extracts of the investigated plants were evaluated against hyperlipidaemia by estimating the rate limiting enzyme of cholesterol biothenysis; β-hydroxy-β-methylglutaryl coenzyme A reductase (HMG-CoA reductase). The antioxidant activity was evaluated by reduction of DPPH(-) free radical. Extra phytochemical screening of Ficus extracts was undertaken, which recorded potent hypolipidaemic and antioxidant activities. The more pronounced extract, Ficus mysorensis (hexane extract), was evaluated in vivo by estimation of the lipid profile and certain antioxidant parameters in hypercholesterolemic rats. The hexane fraction was chromatographed and six isolated compounds were identified. Furthermore, its saponifiable fraction was identified by a MS/MS technique. In conclusion, F. mysorensis recorded hypolipidaemic and antioxidant effects. Detailed studies of the isolated compounds must be undertaken for an evaluation against hypercholesterolemia and free radical elevation.

  5. Toward the discovery of vaccine adjuvants: coupling in silico screening and in vitro analysis of antagonist binding to human and mouse CCR4 receptors.

    Directory of Open Access Journals (Sweden)

    Matthew N Davies

    2009-11-01

    Full Text Available Adjuvants enhance or modify an immune response that is made to an antigen. An antagonist of the chemokine CCR4 receptor can display adjuvant-like properties by diminishing the ability of CD4+CD25+ regulatory T cells (Tregs to down-regulate immune responses.Here, we have used protein modelling to create a plausible chemokine receptor model with the aim of using virtual screening to identify potential small molecule chemokine antagonists. A combination of homology modelling and molecular docking was used to create a model of the CCR4 receptor in order to investigate potential lead compounds that display antagonistic properties. Three-dimensional structure-based virtual screening of the CCR4 receptor identified 116 small molecules that were calculated to have a high affinity for the receptor; these were tested experimentally for CCR4 antagonism. Fifteen of these small molecules were shown to inhibit specifically CCR4-mediated cell migration, including that of CCR4(+ Tregs.Our CCR4 antagonists act as adjuvants augmenting human T cell proliferation in an in vitro immune response model and compound SP50 increases T cell and antibody responses in vivo when combined with vaccine antigens of Mycobacterium tuberculosis and Plasmodium yoelii in mice.

  6. In vitro thrombolytic, anthelmintic, anti-oxidant and cytotoxic activity with phytochemical screening of methanolic extract of Xanthium indicum leaves

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    Antara Ghosh

    2015-12-01

    Full Text Available Xanthium indicum is an important medicinal plant traditionally used in Bangladesh as a folkloric treatment. The current study was undertaken to evaluate thrombolytic, anthelmintic, anti-oxidant, cytotoxic properties with phytochemical screening of methanolic extract of X. indicum leaves. The analysis of phytochemical screening confirmed the existence of phytosetrols and diterpenes. In thrombolytic assay, a significant clot lysis was observed at four concentrations of plant extract compare to the positive control streptokinase (30,000 IU, 15,000 IU and negative control normal saline. The extract revealed potent anthelmintic activity at different concentrations. In anti-oxidant activity evaluation by two potential experiments namely total phenolic content determination and free radical scavenging assay by 2, 2-diphenylpicrylhydrazyl (DPPH, the leaves extract possess good anti-oxidant property. In the brine shrimp lethality bioassay, the crude extract showed potent (LC50 1.3 μg/mL cytotoxic activity compare to the vincristine sulfate as a positive control (LC50 0.8 μg/mL.

  7. Aneuploidy screening by array comparative genomic hybridization improves success rates of in vitro fertilization: A multicenter Indian study

    Directory of Open Access Journals (Sweden)

    Aditi Kotdawala

    2016-01-01

    Full Text Available Objective: To evaluate the usefulness of preimplantation genetic screening (PGS using array comparative genomic hybridization (aCGH in the Indian population. Materials and Methods: This is a retrospective, multicenter study including 235 PGS cycles following intracytoplasmic sperm injection performed at six different infertility centers from September 2013 to June 2015. Patients were divided as per maternal age in several groups (40 years and as per indication for undergoing PGS. Indications for performing PGS were recurrent miscarriage, repetitive implantation failure, severe male factor, previous trisomic pregnancy, and advanced maternal age (≥35. Day 3 embryo biopsy was performed and analyzed by aCGH followed by day 5 embryo transfer in the same cycle or the following cycle. Outcomes such as pregnancy rates (PRs/transfer, implantation rates, miscarriage rates, percentage of abnormal embryos, and number of embryos with more than one aneuploidy and chaotic patterns were recorded for all the treated subjects based on different age and indication groups. Results: aCGH helped in identifying aneuploid embryos, thus leading to consistent implantation (range: 33.3%-42.9% and PRs per transfer (range: 31.8%-54.9% that were obtained for all the indications in all the age groups, after performing PGS. Conclusion: Aneuploidy is one of the major factors which affect embryo implantation. aCGH can be successfully employed for screening of aneuploid embryos. When euploid embryos are transferred, an increase in PRs can be achieved irrespective of the age or the indication.

  8. Identification of natural compound inhibitors for multidrug efflux pumps of Escherichia coli and Pseudomonas aeruginosa using in silico high-throughput virtual screening and in vitro validation.

    Science.gov (United States)

    Aparna, Vasudevan; Dineshkumar, Kesavan; Mohanalakshmi, Narasumani; Velmurugan, Devadasan; Hopper, Waheeta

    2014-01-01

    Pseudomonas aeruginosa and Escherichia coli are resistant to wide range of antibiotics rendering the treatment of infections very difficult. A main mechanism attributed to the resistance is the function of efflux pumps. MexAB-OprM and AcrAB-TolC are the tripartite efflux pump assemblies, responsible for multidrug resistance in P. aeruginosa and E. coli respectively. Substrates that are more susceptible for efflux are predicted to have a common pharmacophore feature map. In this study, a new criterion of excluding compounds with efflux substrate-like features was used, thereby refining the selection process and enriching the inhibitor identification process. An in-house database of phytochemicals was created and screened using high-throughput virtual screening against AcrB and MexB proteins and filtered by matching with the common pharmacophore models (AADHR, ADHNR, AAHNR, AADHN, AADNR, AAADN, AAADR, AAANR, AAAHN, AAADD and AAADH) generated using known efflux substrates. Phytochemical hits that matched with any one or more of the efflux substrate models were excluded from the study. Hits that do not have features similar to the efflux substrate models were docked using XP docking against the AcrB and MexB proteins. The best hits of the XP docking were validated by checkerboard synergy assay and ethidium bromide accumulation assay for their efflux inhibition potency. Lanatoside C and diadzein were filtered based on the synergistic potential and validated for their efflux inhibition potency using ethidium bromide accumulation study. These compounds exhibited the ability to increase the accumulation of ethidium bromide inside the bacterial cell as evidenced by these increase in fluorescence in the presence of the compounds. With this good correlation between in silico screening and positive efflux inhibitory activity in vitro, the two compounds, lanatoside C and diadzein could be promising efflux pump inhibitors and effective to use in combination therapy against drug

  9. Infrared spectroscopic analysis of human interstitial fluid in vitro and in vivo using FT-IR spectroscopy and pulsed quantum cascade lasers (QCL): Establishing a new approach to non invasive glucose measurement

    Science.gov (United States)

    Pleitez, Miguel; von Lilienfeld-Toal, Hermann; Mäntele, Werner

    2012-01-01

    Interstitial fluid, i.e. the liquid present in the outermost layer of living cells of the skin between the Stratum corneum and the Stratum spinosum, was analyzed by Fourier transform infrared spectroscopy and by infrared spectroscopy using pulsed quantum cascade infrared lasers with photoacoustic detection. IR spectra of simulated interstitial fluid samples and of real samples from volunteers in the 850-1800 cm -1 range revealed that the major components of interstitial fluid are albumin and glucose within the physiological range, with only traces of sodium lactate if at all. The IR absorbance of glucose in interstitial fluid in vivo was probed in healthy volunteers using a setup with quantum cascade lasers and photoacoustic detection previously described [11]. A variation of blood glucose between approx. 80 mg/dl and 250 mg/dl in the volunteers was obtained using the standard oral glucose tolerance test (OGT). At two IR wavelengths, 1054 cm -1 and 1084 cm -1, a reasonable correlation between the photoacoustic signal from the skin and the blood glucose value as determined by conventional glucose test sticks using blood from the finger tip was obtained. The infrared photoacoustic glucose signal (PAGS) may serve as the key for a non-invasive glucose measurement, since the glucose content in interstitial fluid closely follows blood glucose in the time course and in the level (a delay of some minutes and a level of approx. 80-90% of the glucose level in blood). Interstitial fluid is present in skin layers at a depth of only 15-50 μm and is thus within the reach of mid-IR energy in an absorbance measurement. A non-invasive glucose measurement for diabetes patients based on mid-infrared quantum cascade lasers and photoacoustic detection could replace the conventional measurement using enzymatic test stripes and a drop of blood from the finger tip, thus reducing pain and being a cost-efficient alternative for millions of diabetes patients.

  10. Quartz-Containing Ceramic Dusts: In vitro screening of the cytotoxic, genotoxic and pro-inflammatory potential of 5 factory samples

    Energy Technology Data Exchange (ETDEWEB)

    Ziemann, C; Creutzenberg, O [Fraunhofer Institute of Toxicology and Experimental Medicine, Hannover (Germany); Jackson, P [CERAM Research Ltd., Stoke-on-Trent (United Kingdom); Brown, R [TOXSERVICES, Stretton (United Kingdom); Attik, G; Rihn, B H, E-mail: christina.ziemann@item.fraunhofer.d [Nancy-University, Faculte de Pharmacie, Nancy (France)

    2009-02-01

    Inhalation of some respirable crystalline silica (MMAD < approx. 4 mum) leads to inflammatory and malignant diseases. Comprehensive physicochemical/biological data and suitable in vitro/in vivo methods may distinguish between more or less harmful quartz-varieties. Within the European Collective Research Project SILICERAM an in vitro screening battery was established to evaluate cytotoxicity (LDH-release, MTT-assay), genotoxicity (Comet-assay) and pro-inflammatory potential (PGE{sub 2}-liberation, TNF-a mRNA expression) of 5 respirable quartz-containing dusts from ceramic plants: brickwork (BR: 7.8% quartz), tableware granulate/cast (TG/TC: 5.8%/3.1%), tiles (TI: 8.1%), refractory (RF: 3.7%). DQ12 (87% a-quartz) and Al{sub 2}O{sub 3} were used as particulate positive and negative controls, respectively. Primary rat alveolar macrophages and the macrophage cell line NR8383 served as model systems. Aluminium lactate was used as inhibitor of biologically active silica, enabling differentiation of silica- and non-specific toxicity. At 200mug/cm{sup 2} (2h) the dusts did not alter significantly LDH-release (except TC), whereas the MTT-assay demonstrated the mainly quartz-independent rank order: DQ12>RF>TG>Ti>BR>TC>Al{sub 2}O{sub 3}. DNA-damage was maximal for BR and TI followed by DQ12>TG>TC>RF>Al{sub 2}O{sub 3}. All dusts induced PGE{sub 2}-liberation (DQ12>BR>TC>TG>Ti>RF>Al{sub 2}O{sub 3}) at 50mug/cm{sup 2} (4h), but TNF-a mRNA (10mug/cm{sup 2}, 24h) was only increased by DQ12, TG (quartz-dependently), and TC. In conclusion, these in vitro tests were an adequate approach to screen the toxic potential of quartz-containing ceramic dusts, but the quartz-content was too low to differentiate the various quartz-varieties.

  11. In-vitro antimicrobial screening and molecular docking studies of synthesized 2-chloro-N-(4-phenylthiazol-2-ylacetamide derivatives

    Directory of Open Access Journals (Sweden)

    Rahmat Ali

    2015-01-01

    Full Text Available Introduction: Glucosamine-6-phosphate (GlcN6P synthase biosynthetic pathway has been identified as potential targets for the development of new antimicrobial agents. Aim: A series of 2-chloro-N-(42-phenylthiazol-25-ylacetamide derivatives (3a-r was synthesized and evaluated their antimicrobial activity. Materials and Methods: The 2-chloro-N-(Para substituted phenylthiazol-25-yl acetamide (2a-c were synthesized by stirring intermediates (1a-c with 2-chloroacetylchloride in dichloromethane in the presence of K2CO3. The intermediate (2a-c were further reacted with different secondary amine such as pyrrolidine, N-methyl piperazine, N-ethyl piperazine, thiomorpholine, morpholine, piperidine etc in ethanol in presence of TEA Triethylamine (TEA to get desired compounds (3a-r. Compounds were characterized by a spectroscopic technique such as Fourier transform infraredFTIR, 1 H-NMR, 13 C-NMR, and mass spectrometry. The synthesized thiazole derivatives (3a-r were screened for anti-bacterial and anti-fungal activity against Escherichia coli, Staphylococcus aureus NCTC 6571, Pseudomonas aeruginosa NCTC 10662, CandidaC. albicans (MTCC-183, AspergillusA. niger (MTCC 281 NCTC 10418 and AspergillusA. flavus (MTCC 277. Result and Conclusion: The results of anti-bacterial screening revealed that among all the screened compounds, eight compounds viz. 3b, 3c, 3d, 3e, 3i, 3j, 3k, and 3p showed moderate to good anti-bacterial and antifungal activity having minimum inhibitory concentration (MIC between 6.25- and 25 µg/ml. While compound 3d showed the most promising antibacterial activity against E. coli and S. aureus, while the compound 3j showed promising antifungal activity with MIC value 6.25 µg/ml against C. albicans, A. niger and A. flavus. In addition, all these eight potential molecules were also examined for possible binding on enzyme GlcN6Pglucosamine-6-phosphate synthase by molecular docking studies on (PDB ID 1JXA.

  12. Total antioxidant potential of juices, beverages and hot drinks consumed in Egypt screened by DPPH in vitro assay

    Directory of Open Access Journals (Sweden)

    Ramadan-Hassanien, Mohamed Fawzy

    2008-09-01

    Full Text Available Plant foods contain different classes and types of antioxidants and knowledge of their total antioxidant potential (TAP, which is the cumulative capacity of food components to scavenge free radicals, would be useful for epidemiological purposes.To accomplish this, a variety of fruit juices, hot drinks and beverages commonly consumed in Egypt were analyzed using in vitro DPPH assay. The order of effectiveness of fruit juices in inhibiting free radicals was as follows: red grapes juice > mango juice > guava juice > cocktail juice > pineapple juice > orange juice > cherry juice > apple juice. Among beverages and hot drinks, teas followed by coffees had the greatest TAP. These data confirm grape juice, teas and coffees as good dietary sources of antioxidants.Las plantas comestibles contienen diferentes clases y tipos de antioxidantes y el conocimiento de su potencial antioxidante total (TAP, que es la capacidad acumulativa de los componentes de los alimentos para captar radicales libres, debería ser útil en estudios epidemiológicos. De acuerdo a esto, una variedad de zumos de fruta, bebidas calientes y bebidas consumidas habitualmente en Egipto fueron analizadas usando un ensayo in vitro con DPPH. El orden de efectividad de los zumos de frutas en inhibir los radicales libres fue el siguiente: zumo de uva tinta > zumo de mango > zumo de guayaba > zumo de macedonia de frutas > zumo de piña >zumo de naranja > zumo de cereza > zumo de manzana. Entre las bebidas y bebidas calientes, el té seguido por el café son los que tuvieron mayores TAPs. Estos datos confirman que el zumo de uva, el té y el café son buenas fuentes de antioxidantes.

  13. Monolayers of IEC-18 cells as an in vitro model for screening the passive transcellular and paracellular transport across the intestinal barrier: Comparison of active and passive transport with the human colon carcinoma Caco-2 cell line

    NARCIS (Netherlands)

    Versantvoort, C.H.M.; Ondrewater, R.C.A.; Duizer, E.; Sandt, J.J.M. van de; Gilde, A.J.; Groten, J.P.

    2002-01-01

    Purpose: previous studies have shown that the rat small intestinal cell line IEC-18 provides a size-selective barrier for paracellularly transported hydrophilic macromolecules. In order to determine the utility of IEC-18 cells as an in vitro model to screen the passive paracellular and transcellular

  14. Ultra-high-throughput screening of an in vitro-synthesized horseradish peroxidase displayed on microbeads using cell sorter.

    Directory of Open Access Journals (Sweden)

    Bo Zhu

    Full Text Available The C1a isoenzyme of horseradish peroxidase (HRP is an industrially important heme-containing enzyme that utilizes hydrogen peroxide to oxidize a wide variety of inorganic and organic compounds for practical applications, including synthesis of fine chemicals, medical diagnostics, and bioremediation. To develop a ultra-high-throughput screening system for HRP, we successfully produced active HRP in an Escherichia coli cell-free protein synthesis system, by adding disulfide bond isomerase DsbC and optimizing the concentrations of hemin and calcium ions and the temperature. The biosynthesized HRP was fused with a single-chain Cro (scCro DNA-binding tag at its N-terminal and C-terminal sites. The addition of the scCro-tag at both ends increased the solubility of the protein. Next, HRP and its fusion proteins were successfully synthesized in a water droplet emulsion by using hexadecane as the oil phase and SunSoft No. 818SK as the surfactant. HRP fusion proteins were displayed on microbeads attached with double-stranded DNA (containing the scCro binding sequence via scCro-DNA interactions. The activities of the immobilized HRP fusion proteins were detected with a tyramide-based fluorogenic assay using flow cytometry. Moreover, a model microbead library containing wild type hrp (WT and inactive mutant (MUT genes was screened using fluorescence-activated cell-sorting, thus efficiently enriching the WT gene from the 1:100 (WT:MUT library. The technique described here could serve as a novel platform for the ultra-high-throughput discovery of more useful HRP mutants and other heme-containing peroxidases.

  15. Identification of β-hematin inhibitors in a high-throughput screening effort reveals scaffolds with in vitro antimalarial activity

    Directory of Open Access Journals (Sweden)

    Rebecca D. Sandlin

    2014-12-01

    Full Text Available The emergence of drug resistant strains of Plasmodium spp. creates a critical need for the development of novel antimalarials. Formation of hemozoin, a crystalline heme detoxification process vital to parasite survival serves as an important drug target. The quinoline antimalarials including chloroquine and amodiaquine owe their antimalarial activity to inhibition of hemozoin formation. Though in vivo formation of hemozoin occurs within the presence of neutral lipids, the lipophilic detergent NP-40 was previously shown to serve as a surrogate in the β-hematin (synthetic hemozoin formation process. Consequently, an NP-40 mediated β-hematin formation assay was developed for use in high-throughput screening. Here, the assay was utilized to screen 144,330 compounds for the identification of inhibitors of crystallization, resulting in 530 hits. To establish the effectiveness of these target-based β-hematin inhibitors against Plasmodium falciparum, each hit was further tested in cultures of parasitized red blood cells. This effort revealed that 171 of the β-hematin inhibitors are also active against the parasite. Dose–response data identified 73 of these β-hematin inhibitors have IC50 values ⩽5 μM, including 25 compounds with nanomolar activity against P. falciparum. A scaffold-based analysis of this data identified 14 primary scaffolds that represent 46% of the 530 total hits. Representative compounds from each of the classes were further assessed for hemozoin inhibitory activity in P. falciparum infected human erythrocytes. Each of the hit compounds tested were found to be positive inhibitors, while a negative control did not perturb this biological pathway in culture.

  16. Functionalized Carbon Quantum Dots with Dopamine for Tyrosinase Activity Monitoring and Inhibitor Screening: In Vitro and Intracellular Investigation.

    Science.gov (United States)

    Chai, Lujing; Zhou, Jin; Feng, Hui; Tang, Cong; Huang, Yuanyuan; Qian, Zhaosheng

    2015-10-28

    Sensitive assay of tyrosinase (TYR) activity is in urgent demand for both fundamental research and practical application, but the exploration of functional materials with good biocompatibility for its activity evaluation at the intracellular level is still challenging until now. In this work, we develop a convenient and real-time assay with high sensitivity for TYR activity/level monitoring and its inhibitor screening based on biocompatible dopamine functionalized carbon quantum dots (Dopa-CQDs). Dopamine with redox property was functionalized on the surface of carbon quantum dots to construct a Dopa-CQDs conjugate with strong bluish green fluorescence. When the dopamine moiety in Dopa-CQDs conjugate was oxidized to a dopaquinone derivative under specific catalysis of TYR, an intraparticle photoinduced electron transfer (PET) process between CQDs and dopaquinone moiety took place, and then the fluorescence of the conjugate could be quenched simultaneously. Quantitative evaluation of TYR activity was established in terms of the relationship between fluorescence quenching efficiency and TYR activity. The assay covered a broad linear range of up to 800 U/L with a low detection limit of 7.0 U/L. Arbutin, a typical inhibitor of TYR, was chosen as an example to assess its function of inhibitor screening, and positive results were observed that fluorescence quenching extent of the probe was reduced in the presence of arbutin. It is also demonstrated that Dopa-CQD conjugate possesses excellent biocompatibility, and can sensitively monitor intracellular tyrosinase level in melanoma cells and intracellular pH changes in living cells, which provides great potential in application of TYR/pH-associated disease monitoring and medical diagnostics.

  17. An in vitro AChE inhibition assay combined with UF-HPLC-ESI-Q-TOF/MS approach for screening and characterizing of AChE inhibitors from roots of Coptis chinensis Franch.

    Science.gov (United States)

    Zhao, Hengqiang; Zhou, Siduo; Zhang, Minmin; Feng, Jinhong; Wang, Shanshan; Wang, Daijie; Geng, Yanling; Wang, Xiao

    2016-02-20

    In this study, an in vitro acetylcholinesterase (AChE) inhibition assay based on microplate reader combined with ultrafiltration high performance liquid chromatography-electrospray quadrupole time of flight mass (UF-HPLC-ESI-Q-TOF/MS) was developed for the rapid screening and identification of acetylcholinesterase inhibitors (AChEI) from roots of Coptis chinensis Franch. Incubation conditions such as enzyme concentration, incubation time, incubation temperature and co-solvent was optimized so as to get better screening results. Five alkaloids including columbamine, jatrorrhizine, coptisine, palmatine and berberine were found with AChE inhibition activity in the 80% ethanol extract of C. chinensis Franch. The screened compounds were identified by HPLC-DAD-ESI-Q-TOF/MS compared with the reference stands and literatures. The screened results were verified by in vitro AChE inhibition assays, palmatine showed the best AChE inhibitory activities with IC50 values of 36.6μM among the five compounds. Results of the present study indicated that the combinative method using in vitro AChE inhibition assay and UF-HPLC-ESI-Q-TOF/MS could be widely applied for rapid screening and identification of AChEI from complex TCM extract. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Transgenic Plasmodium parasites stably expressing Plasmodium vivax dihydrofolate reductase-thymidylate synthase as in vitro and in vivo models for antifolate screening

    Directory of Open Access Journals (Sweden)

    Yuthavong Yongyuth

    2011-10-01

    Full Text Available Abstract Background Plasmodium vivax is the most prevalent cause of human malaria in tropical regions outside the African continent. The lack of a routine continuous in vitro culture of this parasite makes it difficult to develop specific drugs for this disease. To facilitate the development of anti-P. vivax drugs, bacterial and yeast surrogate models expressing the validated P. vivax target dihydrofolate reductase-thymidylate synthase (DHFR-TS have been generated; however, they can only be used as primary screening models because of significant differences in enzyme expression level and in vivo drug metabolism between the surrogate models and P. vivax parasites. Methods Plasmodium falciparum and Plasmodium berghei parasites were transfected with DNA constructs bearing P. vivax dhfr-ts pyrimethamine sensitive (wild-type and pyrimethamine resistant (mutant alleles. Double crossover homologous recombination was used to replace the endogenous dhfr-ts of P. falciparum and P. berghei parasites with P. vivax homologous genes. The integration of Pvdhfr-ts genes via allelic replacement was verified by Southern analysis and the transgenic parasites lines validated as models by standard drug screening assays. Results Transgenic P. falciparum and P. berghei lines stably expressing PvDHFR-TS replacing the endogenous parasite DHFR-TS were obtained. Anti-malarial drug screening assays showed that transgenic parasites expressing wild-type PvDHFR-TS were pyrimethamine-sensitive, whereas transgenic parasites expressing mutant PvDHFR-TS were pyrimethamine-resistant. The growth and sensitivity to other types of anti-malarial drugs in the transgenic parasites were otherwise indistinguishable from the parental parasites. Conclusion With the permanent integration of Pvdhfr-ts gene in the genome, the transgenic Plasmodium lines expressing PvDHFR-TS are genetically stable and will be useful for screening anti-P. vivax compounds targeting PvDHFR-TS. A similar approach

  19. Cascade Distillation System Development

    Science.gov (United States)

    Callahan, Michael R.; Sargushingh, Miriam; Shull, Sarah

    2014-01-01

    NASA's Advanced Exploration Systems (AES) Life Support System (LSS) Project is chartered with de-veloping advanced life support systems that will ena-ble NASA human exploration beyond low Earth orbit (LEO). The goal of AES is to increase the affordabil-ity of long-duration life support missions, and to re-duce the risk associated with integrating and infusing new enabling technologies required to ensure mission success. Because of the robust nature of distillation systems, the AES LSS Project is pursuing develop-ment of the Cascade Distillation Subsystem (CDS) as part of its technology portfolio. Currently, the system is being developed into a flight forward Generation 2.0 design.

  20. In vitro screening for cestocidal activity of three species of Cassia plants against the tapeworm Raillietina tetragona.

    Science.gov (United States)

    Kundu, S; Lyndem, L M

    2013-06-01

    Different species of Cassia plant are widely available in India and are commonly used either for their laxative, antimicrobial or antibacterial activity. In the present study the effectiveness in vitro of the crude alcoholic extracts of three species, namely Cassia alata, C. occidentalis and C. angustifolia, in the early paralysis and mortality of the fowl tapeworm Raillietina tetragona at concentrations ranging from 5 to 80 mg/ml was investigated. Time of paralysis and death were monitored frequently. Immediately after paralysis the tapeworms were processed for electron microscopic studies. While the untreated or control parasites survived for 81.93 ± 5.85 h, the parasites treated with C. alata took less time (1.68 ± 0.27 h) to be paralysed, followed by those treated with C. angustifolia (2.95 ± 0.29 h). Although C. occidentalis took more time (4.13 ± 0.31 h) to paralyse, in combination with either C. alata or C. angustifolia the time taken to paralyse became shorter. All the plant-treated parasites showed irrevocable changes in the scolex and proglottids as compared with the control, and these observations are comparable with those obtained with praziquantel. These results indicate that the three plants tested can be claimed to have anthelmintic activity in addition to their known properties, both when used individually and in combination. Further investigations will be required to evaluate their mechanism of action.

  1. In vitro method for the screening and monitoring of estrogen-deficiency osteoporosis by targeting peripheral circulating monocytes.

    Science.gov (United States)

    Salamanna, Francesca; Maglio, Melania; Giavaresi, Gianluca; Pagani, Stefania; Giardino, Roberto; Fini, Milena

    2015-08-01

    Bone loss occurs insidiously and initially asymptomatically; therefore, osteoporosis is frequently diagnosed only after the first clinical fracture. The aim of this study was to test the hypothesis is that by simply observing the behavior of cultured peripheral monocytes, it might be possible to diagnose altered bone remodeling and, therefore, limit the complications associated with osteoporosis, especially fractures. Monocytes isolated as mononuclear precursors from healthy and ovariectomized rats were cultured both in basal and differentiation medium for up to 3 weeks. Viability and differentiation capability towards the osteoclastic phenotype was checked by light microscopy at early times, whereas differentiation state and synthetic activity (tartrate-resistant acid phosphatase (TRAP) staining; phalloidin, fluorescin isothiocynate (FITC) staining, cathepsin K, metalloproteinase 7 and 9, MMP-7 and MMP-9) were measured at 1, 2, and 3 weeks. Compared to their controls, monocytes isolated from ovariectomized rats proliferate and lean toward the osteoclastic phenotype in the absence of differentiating factors. In both culture conditions, osteoclasts from ovariectomized rats showed significantly higher productions of cathepsin K, MMP-7, and MMP-9 than those of cells isolated from healthy rats, steadily over time. These results obtained in an animal osteoporotic model, if confirmed by clinical studies, open up the possibility to assess the presence of an alteration in bone remodeling with a simple in vitro diagnostic test requiring a small blood sample and less than 48 h. This might allow to early select patients with a spontaneous viability and differentiation of monocytes to osteoclasts for further diagnostic techniques.

  2. In vitro screening and evaluation of antivenom phytochemicals from Azima tetracantha Lam. leaves against Bungarus caeruleus and Vipera russelli.

    Science.gov (United States)

    Janardhan, Bhavya; Shrikanth, Vineetha M; Mirajkar, Kiran K; More, Sunil S

    2014-04-01

    Snakebites are considered a neglected tropical disease that affects thousands of people worldwide. Although antivenom is the only treatment available, it is associated with several side effects. As an alternative, plants have been extensively studied in order to obtain an alternative treatment. In folk medicine, Azima tetracantha Lam. is usually used to treat snakebites. The present study aims to provide a scientific explanation for the use of this plant against snakebite. The extracts of shade dried leaves of A. tetracantha were tested for in vitro inhibitory activity on toxic venom enzymes like phosphomonoesterase, phosphodiesterase, acetylcholinesterase, hyaluronidase etc. from Bungarus caeruleus and Vipera russelli venoms. The ethylacetate extract rendered a significant inhibitory effect on the phosphomonoesterase, phosphodiesterase, phospholipase A2 and acetylcholinesterase enzymes. The present study suggests that ethylacetate extract of A. tetracantha leaves possesses compounds that inhibit the activity of toxic enzymes from Bungarus caeruleus and Vipera russelli venom. Further pharmacological and in vivo studies would provide evidence that this substance may lead to a potential treatment against these venoms.

  3. Screening and In Vitro Evaluation of Mucoadhesive Thermoresponsive System Containing Methylene Blue for Local Photodynamic Therapy of Colorectal Cancer.

    Science.gov (United States)

    Borghi-Pangoni, Fernanda Belincanta; Junqueira, Mariana Volpato; de Souza Ferreira, Sabrina Barbosa; Silva, Larissa Lachi; Rabello, Bruno Ribeiro; Caetano, Wilker; Diniz, Andrea; Bruschi, Marcos Luciano

    2016-03-01

    Photodynamic therapy (PDT) with methylene blue (MB) constitutes a potentially useful modality for colorectal cancer treatment. The limitations of the formulations containing MB are problems of administration and the inability to get the closeness contact at the site during the appropriate residence time. Present study aimed to develop and characterize mucoadhesive thermoresponsive system containing MB designed as platform for colorectal cancer therapy. Formulations composed of different amounts of poloxamer 407 (Polox), Carbopol 934P (Carb), and MB were developed and characterized as rheological, compressional, mucoadhesive and syringeability properties, toxicity, photodynamic action, in vitro MB release profile, and ex vivo MB intestinal permeation. The different compositions resulted in formulations with distinctive macroscopic characteristics and wide range of gelation temperatures. The compressional flow, mucoadhesive, syringeability, and rheological properties were significantly influenced by temperature and/or composition. The MB release from formulation was governed by anomalous transport. In addition, it was observed that MB permeated the intestinal membrane; the formulation possesses photodynamic activity and low toxicity. The data obtained from the system composed of 20% Polox, 0.15% Carb, and 0.25% MB indicated a potentially functional role in PDT of the colorectal cancer and suggest it is worthy of clinical evaluation.

  4. Studying inhibition of calcium oxalate stone formation: an in vitro approach for screening hydrogen sulfide and its metabolites

    Directory of Open Access Journals (Sweden)

    S. Vaitheeswari

    2015-06-01

    Full Text Available ABSTRACTPurpose:Calcium oxalate urolithiasis is one of the most common urinary tract diseases and is of high prevalence. The present study proposes to evaluate the antilithiatic property of hydrogen sulfide and its metabolites like thiosulfate & sulfate in an in vitro model.Materials and Methods:The antilithiatic activity of sodium hydrogen sulfide (NaSH, sodium thiosulfate (Na2S2O3 and sodium sulfate (Na2SO4 on the kinetics of calcium oxalate crystal formation was investigated both in physiological buffer and in urine from normal and recurrent stone forming volunteers. The stones were characterized by optical and spectroscopic techniques.Results:The stones were characterized to be monoclinic, prismatic and bipyramidal habit which is of calcium monohydrate and dihydrate nature. The FTIR displayed fingerprint corresponding to calcium oxalate in the control while in NaSH treated, S=O vibrations were visible in the spectrum. The order of percentage inhibition was NaSH>Na2S2O3>Na2SO4.Conclusion:Our study indicates that sodium hydrogen sulfide and its metabolite thiosulfate are inhibitors of calcium oxalate stone agglomeration which makes them unstable both in physiological buffer and in urine. This effect is attributed to pH changes and complexing of calcium by S2O32-and SO42- moiety produced by the test compounds.

  5. Unsteady turbulence cascades.

    Science.gov (United States)

    Goto, Susumu; Vassilicos, J C

    2016-11-01

    We have run a total of 311 direct numerical simulations (DNSs) of decaying three-dimensional Navier-Stokes turbulence in a periodic box with values of the Taylor length-based Reynolds number up to about 300 and an energy spectrum with a wide wave-number range of close to -5/3 power-law dependence at the higher Reynolds numbers. On the basis of these runs, we have found a critical time when (i) the rate of change of the square of the integral length scale turns from increasing to decreasing, (ii) the ratio of interscale energy flux to high-pass filtered turbulence dissipation changes from decreasing to very slowly increasing in the inertial range, (iii) the signature of large-scale coherent structures disappears in the energy spectrum, and (iv) the scaling of the turbulence dissipation changes from the one recently discovered in DNSs of forced unsteady turbulence and in wind tunnel experiments of turbulent wakes and grid-generated turbulence to the classical scaling proposed by G. I. Taylor [Proc. R. Soc. London, Ser. A 151, 421 (1935)1364-502110.1098/rspa.1935.0158] and A. N. Kolmogorov [Dokl. Akad. Nauk SSSR 31, 538 (1941)]. Even though the customary theoretical basis for this Taylor-Kolmogorov scaling is a statistically stationary cascade where large-scale energy flux balances dissipation, this is not the case throughout the entire time range of integration in all our DNS runs. The recently discovered dissipation scaling can be reformulated physically as a situation in which the dissipation rates of the small and large scales evolve together. We advance two hypotheses that may form the basis of a theoretical approach to unsteady turbulence cascades in the presence of large-scale coherent structures.

  6. In search for geroprotectors: in silico screening and in vitro validation of signalome-level mimetics of young healthy state.

    Science.gov (United States)

    Aliper, Alexander; Belikov, Aleksey V; Garazha, Andrew; Jellen, Leslie; Artemov, Artem; Suntsova, Maria; Ivanova, Alena; Venkova, Larisa; Borisov, Nicolas; Buzdin, Anton; Mamoshina, Polina; Putin, Evgeny; Swick, Andrew G; Moskalev, Alexey; Zhavoronkov, Alex

    2016-09-24

    Populations in developed nations throughout the world are rapidly aging, and the search for geroprotectors, or anti-aging interventions, has never been more important. Yet while hundreds of geroprotectors have extended lifespan in animal models, none have yet been approved for widespread use in humans. GeroScope is a computational tool that can aid prediction of novel geroprotectors from existing human gene expression data. GeroScope maps expression differences between samples from young and old subjects to aging-related signaling pathways, then profiles pathway activation strength (PAS) for each condition. Known substances are then screened and ranked for those most likely to target differential pathways and mimic the young signalome. Here we used GeroScope and shortlisted ten substances, all of which have lifespan-extending effects in animal models, and tested 6 of them for geroprotective effects in senescent human fibroblast cultures. PD-98059, a highly selective MEK1 inhibitor, showed both life-prolonging and rejuvenating effects. Natural compounds like N-acetyl-L-cysteine, Myricetin and Epigallocatechin gallate also improved several senescence-associated properties and were further investigated with pathway analysis. This work not only highlights several potential geroprotectors for further study, but also serves as a proof-of-concept for GeroScope, Oncofinder and other PAS-based methods in streamlining drug prediction, repurposing and personalized medicine.

  7. Characterization of estrogenic and androgenic activity of phthalates by the XenoScreen YES/YAS in vitro assay.

    Science.gov (United States)

    Czernych, Radosław; Chraniuk, Milena; Zagożdżon, Paweł; Wolska, Lidia

    2017-07-01

    The presented study investigates and compares the estrogenic and androgenic activities of commonly used diesters of phthalic acid (phthalates) using the XenoScreen YES/YAS assay. Phthalates are commonly used plasticizers in polymers dedicated for i.e. food and drug containers. Since phthalates are not chemically bonded to the polymer, they can leach or migrate from the polymer. Therefore, phthalates are identified as contaminants in a variety of consumer products. Investigation of estrogenic and androgenic activities of phthalates (DEP, DBP, BBP, DEHP and DINP) showed no significant effect of tested substances either on hERα or hAR receptors. Phthalates exhibited strong anti-estrogenic (IC50 for BBP=8.66μM, IC50 for DEHP=3.61μM and IC50 for DINP=0.065μM) and anti-androgenic (IC50 for BBP=5.30μM, IC50 for DEHP=2.87μM and IC50 for DINP=0.068μM) activities. Copyright © 2017. Published by Elsevier B.V.

  8. Chemometrics Optimized Extraction Procedures, Phytosynergistic Blending and in vitro Screening of Natural Enzyme Inhibitors Amongst Leaves of Tulsi, Banyan and Jamun.

    Science.gov (United States)

    De, Baishakhi; Bhandari, Koushik; Singla, Rajeev K; Katakam, Prakash; Samanta, Tanmoy; Kushwaha, Dilip Kumar; Gundamaraju, Rohit; Mitra, Analava

    2015-10-01

    Tulsi, Banyan, and Jamun are popular Indian medicinal plants with notable hypoglycemic potentials. Now the work reports chemo-profiling of the three species with in-vitro screening approach for natural enzyme inhibitors (NEIs) against enzymes pathogenic for type 2 diabetes. Further along with the chemometrics optimized extraction process technology, phyto-synergistic studies of the composite polyherbal blends have also been reported. Chemometrically optimized extraction procedures, ratios of polyherbal composites to achieve phyto-synergistic actions, and in-vitro screening of NEIs amongst leaves of Tulsi, Banyan, and Jamun. The extraction process parameters of the leaves of three plant species (Ficus benghalensis, Syzigium cumini and Ocimum sanctum) were optimized by rotatable central composite design of chemometrics so as to get maximal yield of bio-actives. Phyto-blends of three species were prepared so as to achieve synergistic antidiabetic and antioxidant potentials and the ratios were optimized by chemometrics. Next, for in vitro screening of natural enzyme inhibitors the individual leaf extracts as well as composite blends were subjected to assay procedures to see their inhibitory potentials against the enzymes pathogenic in type 2 diabetes. The antioxidant potentials were also estimated by DPPH radical scavenging, ABTS, FRAP and Dot Blot assay. Considering response surface methodology studies and from the solutions obtained using desirability function, it was found that hydro-ethanolic or methanolic solvent ratio of 52.46 ± 1.6 and at a temperature of 20.17 ± 0.6 gave an optimum yield of polyphenols with minimal chlorophyll leaching. The species also showed the presence of glycosides, alkaloids, and saponins. Composites in the ratios of 1:1:1 and 1:1:2 gave synergistic effects in terms of polyphenol yield and anti-oxidant potentials. All composites (1:1:1, 1:2:1, 2:1:1, 1:1:2) showed synergistic anti-oxidant actions. Inhibitory activities against the

  9. The use of in vitro technologies and high-resolution/accurate-mass LC-MS to screen for metabolites of 'designer' steroids in the equine.

    Science.gov (United States)

    Clarke, Adam; Scarth, James; Teale, Phil; Pearce, Clive; Hillyer, Lynn

    2011-01-01

    Detection of androgenic-anabolic steroid abuse in equine sports requires knowledge of the drug's metabolism in order to target appropriate metabolites, especially where urine is the matrix of choice. Studying 'designer' steroid metabolism is problematic since it is difficult to obtain ethical approval for in vivo metabolism studies due to a lack of toxicological data. In this study, the equine in vitro metabolism of eight steroids available for purchase on the Internet is reported; including androsta-1,4,6-triene-3,17-dione, 4-chloro,17α-methyl-androsta-1,4-diene-3,17β-diol, estra-4,9-diene-3,17-dione, 4-hydroxyandrostenedione, 20-hydroxyecdysone, 11-keto-androstenedione, 17α-methyldrostanolone, and tetrahydrogestrinone. In order to allow for retrospective analysis of sample testing data, the use of a high-resolution (HR) accurate-mass Thermo LTQ-Orbitrap liquid chromatography-mass spectrometry (LC-MS) instrument was employed for metabolite identification of underivatized sample extracts. The full scan LC-HRMS Orbitrap data were complimented by LC-HRMS/MS and gas-chromatography-mass spectrometry (GC-MS) experiments in order to provide fragmentation information and to ascertain whether GC-MS was capable of detecting any metabolite not detected by LC-HRMS. With the exception of 20-hydroxyecdysone, all compounds were found to be metabolized by equine liver S9 and/or microsomes. With the exception of 17α-methyldrostanolone, which produced metabolites that could only be detected by GC-MS, the metabolites of all other compounds could be identified using LC-HRMS, thus allowing retrospective analysis of previously acquired full-scan data resulting from routine equine drug testing screens. In summary, while in vitro techniques do not serve as a replacement for more definitive in vivo studies in all situations, their use does offer an alternative in situations where it would not be ethical to administer untested drugs to animals.

  10. In vitro screen of a small molecule inhibitor drug library identifies multiple compounds that synergize with oncolytic myxoma virus against human brain tumor-initiating cells.

    Science.gov (United States)

    McKenzie, Brienne A; Zemp, Franz J; Pisklakova, Alexandra; Narendran, Aru; McFadden, Grant; Lun, Xueqing; Kenchappa, Rajappa S; Kurz, Ebba U; Forsyth, Peter A

    2015-08-01

    Brain tumor-initiating cells (BTICs) are stem-like cells hypothesized to form a disease reservoir that mediates tumor recurrence in high-grade gliomas. Oncolytic virotherapy uses replication-competent viruses to target and kill malignant cells and has been evaluated in clinic for glioma therapy with limited results. Myxoma virus (MyxV) is a safe and highly effective oncolytic virus (OV) in conventional glioma models but, as seen with other OVs, is only modestly effective for patient-derived BTICs. The objective of this study was to determine whether MyxV treatment against human BTICs could be improved by combining chemotherapeutics and virotherapy. A 73-compound library of drug candidates in clinical use or preclinical development was screened to identify compounds that sensitize human BTICs to MyxV treatment in vitro, and synergy was evaluated mathematically in lead compounds using Chou-Talalay analyses. The effects of combination therapy on viral gene expression and viral replication were also assessed. Eleven compounds that enhance MyxV efficacy were identified, and 6 were shown to synergize with the virus using Chou-Talalay analyses. Four of the synergistic compounds were shown to significantly increase viral gene expression, indicating a potential mechanism for synergy. Three highly synergistic compounds (axitinib, a VEGFR inhibitor; rofecoxib, a cyclooxygenase-2 inhibitor; and pemetrexed, a folate anti-metabolite) belong to classes of compounds that have not been previously shown to synergize with oncolytic viruses in vitro. This study has identified multiple novel drug candidates that synergistically improve MyxV efficacy in a preclinical BTIC glioma model. © The Author(s) 2015. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Screening of bacterial direct-fed microbials for their antimethanogenic potential in vitro and assessment of their effect on ruminal fermentation and microbial profiles in sheep.

    Science.gov (United States)

    Jeyanathan, J; Martin, C; Morgavi, D P

    2016-02-01

    Direct-fed microbials (DFM) are used to modulate ruminal function and induce beneficial effects on ruminants. The objectives of this work were to 1) screen bacterial strains for their antimethanogenic potential in vitro and 2) assess the effect of 3 selected DFM on ruminal methane (CH) emissions, fermentation parameters, and microbial profiles in sheep. Forty-five bacterial strains were preselected based on their metabolism and fermentation characteristics. These bacteria were screened for their ability to reduce ruminal methanogenesis using 24-h batch incubations and an inoculum of 10 cfu/mL of medium. The addition of bacterial strains stimulated ruminal fermentation with increases in total gas production for 41 strains ( Methane production was reduced by 13% ( < 0.05) with after 2 wk of DFM administration, and this effect was maintained throughout the treatment and posttreatment periods. In contrast, had no effect on CH production, and increased it by 16% ( < 0.05) after 4 wk of DFM administration. There was no effect on other fermentation parameters or on the bacterial, archaeal, and protozoal numbers monitored by quantitative PCR. However, denaturing gradient gel electrophoresis profiles indicated changes in bacterial and archaeal diversity in the and groups. Although added bacteria were unable to permanently colonize the rumen, had a greater 24-h survival rate than the others, implying that the persistence of DFM may be important for modulating ruminal traits of interest. These results suggest that bacterial DFM used in this trial were able to modify CH emissions, although correlated changes in other ruminal parameters studied were minor.

  12. Tyrosine kinase syk non-enzymatic inhibitors and potential anti-allergic drug-like compounds discovered by virtual and in vitro screening.

    Directory of Open Access Journals (Sweden)

    Bruno O Villoutreix

    Full Text Available In the past decade, the spleen tyrosine kinase (Syk has shown a high potential for the discovery of new treatments for inflammatory and autoimmune disorders. Pharmacological inhibitors of Syk catalytic site bearing therapeutic potential have been developed, with however limited specificity towards Syk. To address this topic, we opted for the design of drug-like compounds that could impede the interaction of Syk with its cellular partners while maintaining an active kinase protein. To achieve this challenging task, we used the powerful potential of intracellular antibodies for the modulation of cellular functions in vivo, combined to structure-based in silico screening. In our previous studies, we reported the anti-allergic properties of the intracellular antibody G4G11. With the aim of finding functional mimics of G4G11, we developed an Antibody Displacement Assay and we isolated the drug-like compound C-13, with promising in vivo anti-allergic activity. The likely binding cavity of this compound is located at the close vicinity of G4G11 epitope, far away from the catalytic site of Syk. Here we report the virtual screen of a collection of 500,000 molecules against this new cavity, which led to the isolation of 1000 compounds subsequently evaluated for their in vitro inhibitory effects using the Antibody Displacement Assay. Eighty five compounds were selected and evaluated for their ability to inhibit the liberation of allergic mediators from mast cells. Among them, 10 compounds inhibited degranulation with IC₅₀ values ≤ 10 µM. The most bioactive compounds combine biological activity, significant inhibition of antibody binding and strong affinity for Syk. Moreover, these molecules show a good potential for oral bioavailability and are not kinase catalytic site inhibitors. These bioactive compounds could be used as starting points for the development of new classes of non-enzymatic inhibitors of Syk and for drug discovery endeavour in the

  13. Screening of multi-targeted natural compounds for receptor tyrosine kinases inhibitors and biological evaluation on cancer cell lines, in silico and in vitro.

    Science.gov (United States)

    Singh, Pushpendra; Bast, Felix

    2015-09-01

    Receptors for growth factors encompass within the superfamily of receptor tyrosine kinases and are known to regulate numerous biological processes including cellular growth, proliferation, metabolism, survival, cell differentiation and apoptosis. These receptors have recently caught the attention of the researchers as an attractive target to combat cancer owing to the evidence suggesting their over-expression in cancer cells. Therefore, we studied receptor-based molecular docking of IR (PDB; 3ETA), IGF1R (PDB; 1K3A), EGFR (PDB; 1M17), VEGFIR (PDB; 3HNG), and VEGFIIR (PDB; 2OH4) against natural compounds. Further, in vitro investigation of the biological effect of lead molecules in an array of cancer cell lines was done. All selected natural compounds were docked with the X-ray crystal structure of selected protein by employing GLIDE (Grid-based Ligand Docking with Energetics) Maestro 9.6. InterBioScreen natural compounds docked with each selected protein molecules by using GLIDE high throughput virtual screening. On the basis of Gscore, we select 20 compounds along with 68 anticancer compounds for GLIDE extra precision molecular docking. It was discovered in this study that compound epigallocatechin gallate (EGCG) yielded magnificent Gscore with IGF1R (PDB; 1K3A) and VEGFIIR (PDB; 2OH4), and protein-ligand interactions are chart out. Effect of EGCG on biological activity such as mRNA expression of selected protein, cell proliferation, oxidative stress, and cell migration was reported after the 48 h treatments in cancer cell lines. The RT-PCR densitometric bands analysis showed that compound EGCG reduced the mRNA expression of IGF1R, VEGFIIR, and mTOR at 80 μM concentration. Moreover, EGCG significantly reduced cell proliferation and ROS generation after 48 h treatments. Our result also indicated a reduction in the potential for cell migration that might show in vivo anti-metastasis activity of EGCG.

  14. Screening of rhizospheric actinomycetes for various in-vitro and in-vivo plant growth promoting (PGP traits and for agroactive compounds

    Directory of Open Access Journals (Sweden)

    Sumaira Anwar

    2016-08-01

    Full Text Available In this study 98 rhizospheric actinomycetes were isolated from different wheat and tomato fields, Punjab, Pakistan. The isolates were characterized morphologically, biochemically and genetically and were subjected to a comprehensive in vitro screening for various plant growth promoting (PGP traits. About 30% of the isolates screened were found to be the promising plant growth promoting rhizobacteria (PGPRs, which exhibited maximum genetic similarity (up to 98-99% with different species of the genus Streptomyces by using16S rRNA gene sequencing. The most active indole acetic acid (IAA producer Streptomyces nobilis WA-3, Streptomyces Kunmingenesis WC-3 and Streptomyces enissocaesilis TA-3 produce 79.5, 79.23 and 69.26 µg/ml IAA respectively at 500µg/ml L-tryptophan. The highest concentration of soluble phosphate was produced by Streptomyces sp. WA-1 (72.13 mg/100ml and S. djakartensis TB-4 (70.36 mg/100ml. All rhizobacterial isolates were positive for siderophore, ammonia and hydrogen cyanide production. Strain S. mutabilis WD-3 showed highest concentration of ACC-deaminase (1.9 mmol /l. For in-vivo screening, seed germination and plant growth experiment were conducted by inoculating wheat (Triticum aestivum seeds with the six selected isolates. Significant increases in shoot length was observed with S. nobilis WA-3 (65 %, increased root length was recorded in case of S. nobilis WA-3 (81 % as compared to water treated control plants. Maximum increases in plant fresh weight were recorded with S. nobilis WA-3 (84 %, increased plant dry weight was recorded in case of S. nobilis WA-3 (85 % as compared to water treated control plants. In case of number of leaves, significant increase was recorded with S. nobilis WA-3 (27 % and significant increase in case of number of roots were recorded in case of strain S. nobilis WA-3 (30 % as compared to control plants. Over all the study revealed that these rhizospheric plant growth promoting (PGP Streptomyces

  15. Screening of Rhizospheric Actinomycetes for Various In-vitro and In-vivo Plant Growth Promoting (PGP) Traits and for Agroactive Compounds.

    Science.gov (United States)

    Anwar, Sumaira; Ali, Basharat; Sajid, Imran

    2016-01-01

    In this study 98 rhizospheric actinomycetes were isolated from different wheat and tomato fields, Punjab, Pakistan. The isolates were characterized morphologically, biochemically, and genetically and were subjected to a comprehensive in vitro screening for various plant growth promoting (PGP) traits. About 30% of the isolates screened were found to be the promising PGP rhizobacteria (PGPRs), which exhibited maximum genetic similarity (up to 98-99%) with different species of the genus Streptomyces by using16S rRNA gene sequencing. The most active indole acetic acid (IAA) producer Streptomyces nobilis WA-3, Streptomyces Kunmingenesis WC-3, and Streptomyces enissocaesilis TA-3 produce 79.5, 79.23, and 69.26 μg/ml IAA respectively at 500 μg/ml L-tryptophan. The highest concentration of soluble phosphate was produced by Streptomyces sp. WA-1 (72.13 mg/100 ml) and S. djakartensis TB-4 (70.36 mg/100 ml). All rhizobacterial isolates were positive for siderophore, ammonia, and hydrogen cyanide production. Strain S. mutabilis WD-3 showed highest concentration of ACC-deaminase (1.9 mmol /l). For in-vivo screening, seed germination, and plant growth experiment were conducted by inoculating wheat (Triticum aestivum) seeds with the six selected isolates. Significant increases in shoot length was observed with S. nobilis WA-3 (65%), increased root length was recorded in case of S. nobilis WA-3 (81%) as compared to water treated control plants. Maximum increases in plant fresh weight were recorded with S. nobilis WA-3 (84%), increased plant dry weight was recorded in case of S. nobilis WA-3 (85%) as compared to water treated control plants. In case of number of leaves, significant increase was recorded with S. nobilis WA-3 (27%) and significant increase in case of number of roots were recorded in case of strain S. nobilis WA-3 (30%) as compared to control plants. Over all the study revealed that these rhizospheric PGP Streptomyces are good candidates to be developed as

  16. Inferring network structure from cascades

    Science.gov (United States)

    Ghonge, Sushrut; Vural, Dervis Can

    2017-07-01

    Many physical, biological, and social phenomena can be described by cascades taking place on a network. Often, the activity can be empirically observed, but not the underlying network of interactions. In this paper we offer three topological methods to infer the structure of any directed network given a set of cascade arrival times. Our formulas hold for a very general class of models where the activation probability of a node is a generic function of its degree and the number of its active neighbors. We report high success rates for synthetic and real networks, for several different cascade models.

  17. Cascade Mountain Range in Oregon

    Science.gov (United States)

    Sherrod, David R.

    2016-01-01

    The Cascade mountain system extends from northern California to central British Columbia. In Oregon, it comprises the Cascade Range, which is 260 miles long and, at greatest breadth, 90 miles wide (fig. 1). Oregon’s Cascade Range covers roughly 17,000 square miles, or about 17 percent of the state, an area larger than each of the smallest nine of the fifty United States. The range is bounded on the east by U.S. Highways 97 and 197. On the west it reaches nearly to Interstate 5, forming the eastern margin of the Willamette Valley and, farther south, abutting the Coast Ranges. 

  18. A well-refined in vitro model derived from human embryonic stem cell for screening phytochemicals with midbrain dopaminergic differentiation-boosting potential for improving Parkinson's disease.

    Science.gov (United States)

    Hsieh, Wen-Ting; Chiang, Been-Huang

    2014-07-09

    Stimulation of endogenous neurogenesis is a potential approach to compensate for loss of dopaminergic neurons of substantia nigra compacta nigra (SNpc) in patients with Parkinson's disease (PD). This objective was to establish an in vitro model by differentiating pluripotent human embryonic stem cells (hESCs) into midbrain dopaminergic (mDA) neurons for screening phytochemicals with mDA neurogenesis-boosting potentials. Consequently, a five-stage differentiation process was developed. The derived cells expressed many mDA markers including tyrosine hydroxylase (TH), β-III tubulin, and dopamine transporter (DAT). The voltage-gated ion channels and dopamine release were also examined for verifying neuron function, and the dopamine receptor agonists bromocriptine and 7-hydroxy-2-(dipropylamino)tetralin (7-OH-DPAT) were used to validate our model. Then, several potential phytochemicals including green tea catechins and ginsenosides were tested using the model. Finally, ginsenoside Rb1 was identified as the most potent phytochemical which is capable of upregulating neurotrophin expression and inducing mDA differentiation.

  19. Comparing antioxidant capacity of purine alkaloids: a new, efficient trio for screening and discovering potential antioxidants in vitro and in vivo.

    Science.gov (United States)

    Tsoi, Bun; Yi, Ruo-Nan; Cao, Ling-Fang; Li, Shan-Bing; Tan, Rui-Rong; Chen, Min; Li, Xiao-Xiao; Wang, Chen; Li, Yi-Fang; Kurihara, Hiroshi; He, Rong-Rong

    2015-06-01

    The most commonly applied strategies for the evaluation of antioxidant capacity are the chemical- or cell-based approaches. However, the results obtained from these methods might not reflect the antioxidant ability of test samples within organisms. In this study, we propose a combination of experiments, including oxygen radical absorbance capacity (ORAC), cellular antioxidant activity assay (CAA), and the chick embryo model, as an efficient trio to evaluate antioxidant capacity of food components. Taking purine alkaloids as example, results demonstrate that chemical and cellular method might misinterpret their true ability on antioxidation. In chick embryo model, caffeine and theacrine can significantly improve vessel density on chorioallantoic membrane and myocardial apoptosis. The mechanism can be involving multiple targets within the organism. We believe that the trio proposed can be widely utilized in screening massive number of antioxidant in a cost-effective way. It will also help discovering new antioxidants that are easily being omitted due to their relatively poor in vitro activities. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Screening of the in vitro antileishmanial activities of compounds and secondary metabolites isolated from Maytenus guianensis Klotzsch ex Reissek (Celastraceae chichuá Amazon

    Directory of Open Access Journals (Sweden)

    Dionatas Ulises de Oliveira Meneguetti

    Full Text Available Abstract INTRODUCTION Maytenus guianensis is a member of the Celastraceae family that is used in traditional medicine, particularly for its anti-parasitic and anti-cancer effects. To explore the ethnopharmacological potential of this plant, the present study was designed to screen the in vitro antileishmanial activities of extracts and compounds isolated from M. guianensis. METHODS Maytenus guianensis stems and leaves were extracted in acetone, followed by the preparation of eluates and isolation of secondary metabolites using chromatography on a glass column with silica gel as the fixed phase. The chemical components were identified using spectroscopic methods, including one- and two-dimensional nuclear magnetic resonance of hydrogen-1 and carbon-13, mass spectroscopy, and infrared spectroscopy. The anti-Leishmania amazonensis activities of these eluates and compounds were evaluated by direct promastigote counting and viability assays. RESULTS It was found that the hexane bark eluate produced the strongest anti-L. amazonensis effect, with 90-100% inhibition of the promastigote form. The isolated metabolite that produced the best result was tingenone B, followed by a compound formed by the union of tingenone and tingenone B (80-90% inhibition. CONCLUSIONS Maytenus guianensis shows anti-parasite activity that warrants further investigation to determine the mechanisms underlying this antileishmanial effect and to evaluate the pharmacological potential of these eluates and isolated secondary metabolites, while minimizing any adverse effects.

  1. Screening of the in vitro antileishmanial activities of compounds and secondary metabolites isolated from Maytenus guianensis Klotzsch ex Reissek (Celastraceae) chichuá Amazon.

    Science.gov (United States)

    Meneguetti, Dionatas Ulises de Oliveira; Lima, Renato Abreu; Hurtado, Fernanda Bay; Passarini, Guilherme Matos; Macedo, Sharon Rose Aragão; Barros, Neuza Biguinati de; Oliveira, Flávio Augusto de Souza; Medeiros, Patrícia Soares de Maria de; Militão, Júlio Sancho Linhares Teixeira; Nicolete, Roberto; Facundo, Valdir Alves

    2016-01-01

    Maytenus guianensis is a member of the Celastraceae family that is used in traditional medicine, particularly for its anti-parasitic and anti-cancer effects. To explore the ethnopharmacological potential of this plant, the present study was designed to screen the in vitro antileishmanial activities of extracts and compounds isolated from M. guianensis. Maytenus guianensis stems and leaves were extracted in acetone, followed by the preparation of eluates and isolation of secondary metabolites using chromatography on a glass column with silica gel as the fixed phase. The chemical components were identified using spectroscopic methods, including one- and two-dimensional nuclear magnetic resonance of hydrogen-1 and carbon-13, mass spectroscopy, and infrared spectroscopy. The anti-Leishmania amazonensis activities of these eluates and compounds were evaluated by direct promastigote counting and viability assays. It was found that the hexane bark eluate produced the strongest anti-L. amazonensis effect, with 90-100% inhibition of the promastigote form. The isolated metabolite that produced the best result was tingenone B, followed by a compound formed by the union of tingenone and tingenone B (80-90% inhibition). Maytenus guianensis shows anti-parasite activity that warrants further investigation to determine the mechanisms underlying this antileishmanial effect and to evaluate the pharmacological potential of these eluates and isolated secondary metabolites, while minimizing any adverse effects.

  2. Cascading effects following intervention.

    Science.gov (United States)

    Patterson, Gerald R; Forgatch, Marion S; Degarmo, David S

    2010-11-01

    Four different sources for cascade effects were examined using 9-year process and outcome data from a randomized controlled trial of a preventive intervention using the Parent Management Training-Oregon Model (PMTO™). The social interaction learning model of child antisocial behavior serves as one basis for predicting change. A second source addresses the issue of comorbid relationships among clinical diagnoses. The third source, collateral changes, describes events in which changes in one family member correlate with changes in another. The fourth component is based on the long-term effects of reducing coercion and increasing positive interpersonal processes within the family. New findings from the 9-year follow-up show that mothers experienced benefits as measured by standard of living (i.e., income, occupation, education, and financial stress) and frequency of police arrests. It is assumed that PMTO reduces the level of coercion, which sets the stage for a massive increase in positive social interaction. In effect, PMTO alters the family environment and thereby opens doors to healthy new social environments.

  3. Injectorless quantum cascade lasers

    Science.gov (United States)

    Katz, Simeon; Vizbaras, Augustinas; Meyer, Ralf; Amann, Markus-Christian

    2011-04-01

    This review focuses on recent progress on injectorless quantum cascade lasers, an increasingly attractive approach in comparison to the "classical" injectorbased concepts. This particularly holds for the wavelength range between 7 and 12 μm, where fundamental vibrational modes of many important molecules exist, so that sensor systems for medical, industrial and military applications highly benefit from these laser sources. The atmospheric transmission window between 8 and 12 μm, with very low damping, also enables free space applications like communication, military countermeasures, and environmental sensors. Injectorless devices operate closer to the original design principle for intersubband lasers as suggested by Suris and Kazarinov [Sov. Phys. Semicond. 5, 707 (1971)]. Therefore, a short description of their features is given in comparison to injectorbased devices. Within recent years, injectorless devices have seen rapid improvement in performance. Best injectorless devices reach threshold current densities of 450 A/cm2 at 300 K, a factor of 1.6 smaller than that for the best injectorbased devices. Their output efficiency has also increased from 2% to more than 7% within the last 2 years, reaching comparable levels and making the injectorless device concept competitive and very attractive for applications.

  4. Cascade dynamics on complex networks.

    OpenAIRE

    Hackett, Adam W.

    2011-01-01

    peer-reviewed The network topologies on which many natural and synthetic systems are built provide ideal settings for the emergence of complex phenomena. One well-studied manifestation of this, called a cascade or avalanche, is observed when interactions between the components of a system allow an initially localized effect to propagate globally. For example, the malfunction of technological systems like email networks or electrical power grids is often attributable to a cascade o...

  5. Interband Cascade Photovoltaic Cells

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Rui Q. [Univ. of Oklahoma, Norman, OK (United States); Santos, Michael B. [Univ. of Oklahoma, Norman, OK (United States); Johnson, Matthew B. [Univ. of Oklahoma, Norman, OK (United States)

    2014-09-24

    In this project, we are performing basic and applied research to systematically investigate our newly proposed interband cascade (IC) photovoltaic (PV) cells [1]. These cells follow from the great success of infrared IC lasers [2-3] that pioneered the use of quantum-engineered IC structures. This quantum-engineered approach will enable PV cells to efficiently convert infrared radiation from the sun or other heat source, to electricity. Such cells will have important applications for more efficient use of solar energy, waste-heat recovery, and power beaming in combination with mid-infrared lasers. The objectives of our investigations are to: achieve extensive understanding of the fundamental aspects of the proposed PV structures, develop the necessary knowledge for making such IC PV cells, and demonstrate prototype working PV cells. This research will focus on IC PV structures and their segments for utilizing infrared radiation with wavelengths from 2 to 5 μm, a range well suited for emission by heat sources (1,000-2,000 K) that are widely available from combustion systems. The long-term goal of this project is to push PV technology to longer wavelengths, allowing for relatively low-temperature thermal sources. Our investigations address material quality, electrical and optical properties, and their interplay for the different regions of an IC PV structure. The tasks involve: design, modeling and optimization of IC PV structures, molecular beam epitaxial growth of PV structures and relevant segments, material characterization, prototype device fabrication and testing. At the end of this program, we expect to generate new cutting-edge knowledge in the design and understanding of quantum-engineered semiconductor structures, and demonstrate the concepts for IC PV devices with high conversion efficiencies.

  6. Action potential-based MEA platform for in vitro screening of drug-induced cardiotoxicity using human iPSCs and rat neonatal myocytes.

    Science.gov (United States)

    Jans, Danny; Callewaert, Geert; Krylychkina, Olga; Hoffman, Luis; Gullo, Francesco; Prodanov, Dimiter; Braeken, Dries

    2017-09-01

    Drug-induced cardiotoxicity poses a negative impact on public health and drug development. Cardiac safety pharmacology issues urged for the preclinical assessment of drug-induced ventricular arrhythmia leading to the design of several in vitro electrophysiological screening assays. In general, patch clamp systems allow for intracellular recordings, while multi-electrode array (MEA) technology detect extracellular activity. Here, we demonstrate a complementary metal oxide semiconductor (CMOS)-based MEA system as a reliable platform for non-invasive, long-term intracellular recording of cardiac action potentials at high resolution. Quinidine (8 concentrations from 10 -7 to 2.10 -5 M) and verapamil (7 concentrations from 10 -11 to 10 -5 M) were tested for dose-dependent responses in a network of cardiomyocytes. Electrophysiological parameters, such as the action potential duration (APD), rates of depolarization and repolarization and beating frequency were assessed. In hiPSC, quinidine prolonged APD with EC 50 of 2.2·10 -6 M. Further analysis indicated a multifactorial action potential prolongation by quinidine: (1) decreasing fast repolarization with IC 50 of 1.1·10 -6 M; (2) reducing maximum upstroke velocity with IC 50 of 2.6·10 -6 M; and (3) suppressing spontaneous activity with EC 50 of 3.8·10 -6 M. In rat neonatal cardiomyocytes, verapamil blocked spontaneous activity with EC 50 of 5.3·10 -8 M and prolonged the APD with EC 50 of 2.5·10 -8 M. Verapamil reduced rates of fast depolarization and repolarization with IC 50 s of 1.8 and 2.2·10 -7 M, respectively. In conclusion, the proposed action potential-based MEA platform offers high quality and stable long-term recordings with high information content allowing to characterize multi-ion channel blocking drugs. We anticipate application of the system as a screening platform to efficiently and cost-effectively test drugs for cardiac safety. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. In vitro field potential monitoring on a multi-microelectrode array for the electrophysiological long-term screening of neural stem cell maturation.

    Science.gov (United States)

    Seidel, Diana; Jahnke, Heinz-Georg; Englich, Beate; Girard, Mathilde; Robitzki, Andrea A

    2017-05-30

    Due to the lack of appropriate cell models as well as automated electrophysiology monitoring technologies, the standardized identification of neurotoxic or protective effects in vitro remains a major problem in today's pharmaceutical ingredient development. Over the past few years, in vivo-like human pluripotent stem cell-derived neuronal networks have turned out to be a promising physiological cell source, if the establishment of robust and time-saving functional maturation strategies based on stable and expandable neural progenitor populations can be achieved. Here, we describe a multi-microelectrode array (MMEA)-based bioelectronics platform that was optimized for long-term electrophysiological activity monitoring of neuronal networks via field potential measurements. Differentiation of small molecule-based neuronal progenitors on MMEAs led to functional neurons within 15 days. More strikingly, these functional neuronal cultures could remain electrophysiologically stable on the MMEAs for more than four weeks. The observed electrophysiological properties correlated with the expression of typical neuron subtype markers and were further validated by specific neurotransmitter applications. With our established monitoring platform, we could show for the first time the long-term stability of the neural stem cell-like progenitor population to differentiate to electrophysiologically active dopaminergic neuronal networks for more than 80 passages. In conclusion, we provide a comprehensive long-term stable field potential monitoring platform based on stem cell-derived human neuronal networks that can be automated and up-scaled for standardized high-content screening applications e.g. in the field of neurotoxic and neuroprotective therapeutics identification.

  8. Mineralogical analyses and in vitro screening tests for the rapid evaluation of the health hazard of volcanic ash at Rabaul volcano, Papua New Guinea

    Science.gov (United States)

    Le Blond, Jennifer S.; Horwell, Claire J.; Baxter, Peter J.; Michnowicz, Sabina A. K.; Tomatis, Maura; Fubini, Bice; Delmelle, Pierre; Dunster, Christina; Patia, Herman

    2010-11-01

    The continuous ash and gas emissions from the Tavurvur cone in Rabaul caldera, Papua New Guinea, during 2007-08, raised concerns regarding how exposure would affect the respiratory health of nearby populations and impact on the environment. As part of a formal evaluation of the effects of volcanic emissions on public health, we investigated the potential health hazard of the ash using a suite of selected mineralogical analyses and in vitro toxicity screening tests. The trachy -andesitic ash comprised 2.1-6.7 vol.% respirable (sub -4 μm diameter) particles. The crystalline silica content was 1.9-5.0 wt.% cristobalite (in the bulk sample) with trace amounts of quartz and/or tridymite. Scanning electron microscopy showed that the ash particles were angular with sparse, fibre -like particles (˜3-60 μm max. diameter) observed in some samples, which we confirmed to be CaSO4 (gypsum, at tested volcanic ash leachates. Ash samples generated potentially -harmful hydroxyl radicals through an iron -mediated catalytic reaction, in the range of 0.15-2.47 μmol m-2 (after 30 min of reaction). However, measurement of particle oxidative capacity (potential oxidative stress reaction using ascorbic acid) and silica -like injury to red blood cells (erythrolysis assay, i.e. measurement of cell death) nevertheless revealed low biological reactivity. The findings suggest that acute exposure to the ash would have a limited potential to exacerbate pre -existing conditions such as asthma or chronic bronchitis, and the potential for chronic exposure leading to silicosis was low.

  9. Cascaded failures in weighted networks

    Science.gov (United States)

    Mirzasoleiman, Baharan; Babaei, Mahmoudreza; Jalili, Mahdi; Safari, Mohammadali

    2011-10-01

    Many technological networks can experience random and/or systematic failures in their components. More destructive situations can happen if the components have limited capacity, where the failure in one of them might lead to a cascade of failures in other components, and consequently break down the structure of the network. In this paper, the tolerance of cascaded failures was investigated in weighted networks. Three weighting strategies were considered including the betweenness centrality of the edges, the product of the degrees of the end nodes, and the product of their betweenness centralities. Then, the effect of the cascaded attack was investigated by considering the local weighted flow redistribution rule. The capacity of the edges was considered to be proportional to their initial weight distribution. The size of the survived part of the attacked network was determined in model networks as well as in a number of real-world networks including the power grid, the internet in the level of autonomous system, the railway network of Europe, and the United States airports network. We found that the networks in which the weight of each edge is the multiplication of the betweenness centrality of the end nodes had the best robustness against cascaded failures. In other words, the case where the load of the links is considered to be the product of the betweenness centrality of the end nodes is favored for the robustness of the network against cascaded failures.

  10. Supersymmetric cascade decays at NLO

    Energy Technology Data Exchange (ETDEWEB)

    Popenda, Eva; Muehlleitner, Margarete; Hangst, Christian [KIT, Institut fuer Theoretische Physik (Germany); Kraemer, Michael [RWTH Aachen University, Institut fuer Theoretische Teilchenphysik und Kosmologie (Germany); Spira, Michael [Paul Scherrer Institut, Theory Group LTP (Switzerland)

    2012-07-01

    The search for supersymmetric particles and determination of their properties is a major task at the LHC and is based on the analysis of the cascade decay chains in which SUSY particles are produced. This project aims at improving predictions for SUSY cascade decays through the inclusion of higher-order corrections in the production and decay processes and by embedding them in a fully flexible Monte Carlo program. In this talk we report on the progress of the implementation of squark pair production followed by the decay into a quark and the lightest neutralino including supersymmetric QCD corrections at next-to-leading order in a completely differential form.

  11. Bosonic cascades of indirect excitons

    Science.gov (United States)

    Nalitov, A. V.; De Liberato, S.; Lagoudakis, P.; Savvidis, P. G.; Kavokin, A. V.

    2017-08-01

    Recently, the concept of the terahertz bosonic cascade laser (BCL) based on a parabolic quantum well (PQW) embedded in a microcavity was proposed. We refine this proposal by suggesting transitions between indirect exciton (IX) states as a source of terahertz emission. We explicitly propose a structure containing a narrow-square QW and a wide-parabolic QW for the realisation of a bosonic cascade. Advantages of this type of structures are in large dipole matrix elements for terahertz transitions and in long exciton radiative lifetimes which are crucial for realisation of threshold and quantum efficiency BCLs.

  12. Cascaded logic gates in nanophotonic plasmon networks

    National Research Council Canada - National Science Library

    Wei, Hong; Wang, Zhuoxian; Tian, Xiaorui; Käll, Mikael; Xu, Hongxing

    2011-01-01

    ... integrated logic units and cascade devices have not been reported. Here we demonstrate that a plasmonic binary NOR gate, a 'universal logic gate', can be realized through cascaded OR and NOT gates in four-terminal plasmonic nanowire networks...

  13. Cascade Support Vector Machines with Dimensionality Reduction

    Directory of Open Access Journals (Sweden)

    Oliver Kramer

    2015-01-01

    Full Text Available Cascade support vector machines have been introduced as extension of classic support vector machines that allow a fast training on large data sets. In this work, we combine cascade support vector machines with dimensionality reduction based preprocessing. The cascade principle allows fast learning based on the division of the training set into subsets and the union of cascade learning results based on support vectors in each cascade level. The combination with dimensionality reduction as preprocessing results in a significant speedup, often without loss of classifier accuracies, while considering the high-dimensional pendants of the low-dimensional support vectors in each new cascade level. We analyze and compare various instantiations of dimensionality reduction preprocessing and cascade SVMs with principal component analysis, locally linear embedding, and isometric mapping. The experimental analysis on various artificial and real-world benchmark problems includes various cascade specific parameters like intermediate training set sizes and dimensionalities.

  14. Activation Cascading in Sign Production

    Science.gov (United States)

    Navarrete, Eduardo; Peressotti, Francesca; Lerose, Luigi; Miozzo, Michele

    2017-01-01

    In this study, we investigated how activation unfolds in sign production by examining whether signs that are not produced have their representations activated by semantics (cascading of activation). Deaf signers were tested with a picture-picture interference task. Participants were presented with pairs of overlapping pictures and named the green…

  15. Dynamics robustness of cascading systems.

    Directory of Open Access Journals (Sweden)

    Jonathan T Young

    2017-03-01

    Full Text Available A most important property of biochemical systems is robustness. Static robustness, e.g., homeostasis, is the insensitivity of a state against perturbations, whereas dynamics robustness, e.g., homeorhesis, is the insensitivity of a dynamic process. In contrast to the extensively studied static robustness, dynamics robustness, i.e., how a system creates an invariant temporal profile against perturbations, is little explored despite transient dynamics being crucial for cellular fates and are reported to be robust experimentally. For example, the duration of a stimulus elicits different phenotypic responses, and signaling networks process and encode temporal information. Hence, robustness in time courses will be necessary for functional biochemical networks. Based on dynamical systems theory, we uncovered a general mechanism to achieve dynamics robustness. Using a three-stage linear signaling cascade as an example, we found that the temporal profiles and response duration post-stimulus is robust to perturbations against certain parameters. Then analyzing the linearized model, we elucidated the criteria of when signaling cascades will display dynamics robustness. We found that changes in the upstream modules are masked in the cascade, and that the response duration is mainly controlled by the rate-limiting module and organization of the cascade's kinetics. Specifically, we found two necessary conditions for dynamics robustness in signaling cascades: 1 Constraint on the rate-limiting process: The phosphatase activity in the perturbed module is not the slowest. 2 Constraints on the initial conditions: The kinase activity needs to be fast enough such that each module is saturated even with fast phosphatase activity and upstream changes are attenuated. We discussed the relevance of such robustness to several biological examples and the validity of the above conditions therein. Given the applicability of dynamics robustness to a variety of systems, it

  16. Cascaded Bragg scattering in fiber optics.

    Science.gov (United States)

    Xu, Y Q; Erkintalo, M; Genty, G; Murdoch, S G

    2013-01-15

    We report on a theoretical and experimental study of cascaded Bragg scattering in fiber optics. We show that the usual energy-momentum conservation of Bragg scattering can be considerably relaxed via cascade-induced phase-matching. Experimentally we demonstrate frequency translation over six- and 11-fold cascades, in excellent agreement with derived phase-matching conditions.

  17. Correlation between in vitro mucus adhesion and the in vivo colonization ability of lactic acid bacteria: screening of new candidate carp probiotics.

    Science.gov (United States)

    Sugimura, Yuya; Hagi, Tatsuro; Hoshino, Takayuki

    2011-01-01

    We measured the adhesion of candidate probiotic lactic acid bacteria (LAB) to carp intestinal mucus. The percentage of adherent bacteria varied among strains. Four strains, two with high adhesion and two with low adhesion in vitro, were tested for in vivo colonization ability. Carp were fed LAB-containing feed for 12 d, and then unsupplemented feed until day 33, and the numbers and compositions of intestinal LAB were analyzed during the entire period. LAB with lower in vitro adhesion disappeared quickly from the intestine after LAB feeding stopped. LAB with higher in vitro adhesion remained in the intestine 3 weeks after LAB feeding stopped, indicating a strong correlation between mucus adhesion in vitro and colonization ability in vivo. Next we isolated nine candidate probiotic LAB with high in vitro mucus-binding ability. Three of them were fed to carp, and all three were stably maintained in the intestine.

  18. Forward design of a complex enzyme cascade reaction

    Science.gov (United States)

    Hold, Christoph; Billerbeck, Sonja; Panke, Sven

    2016-09-01

    Enzymatic reaction networks are unique in that one can operate a large number of reactions under the same set of conditions concomitantly in one pot, but the nonlinear kinetics of the enzymes and the resulting system complexity have so far defeated rational design processes for the construction of such complex cascade reactions. Here we demonstrate the forward design of an in vitro 10-membered system using enzymes from highly regulated biological processes such as glycolysis. For this, we adapt the characterization of the biochemical system to the needs of classical engineering systems theory: we combine online mass spectrometry and continuous system operation to apply standard system theory input functions and to use the detailed dynamic system responses to parameterize a model of sufficient quality for forward design. This allows the facile optimization of a 10-enzyme cascade reaction for fine chemical production purposes.

  19. Bankruptcy cascades in interbank markets.

    Directory of Open Access Journals (Sweden)

    Gabriele Tedeschi

    Full Text Available We study a credit network and, in particular, an interbank system with an agent-based model. To understand the relationship between business cycles and cascades of bankruptcies, we model a three-sector economy with goods, credit and interbank market. In the interbank market, the participating banks share the risk of bad debits, which may potentially spread a bank's liquidity problems through the network of banks. Our agent-based model sheds light on the correlation between bankruptcy cascades and the endogenous economic cycle of booms and recessions. It also demonstrates the serious trade-off between, on the one hand, reducing risks of individual banks by sharing them and, on the other hand, creating systemic risks through credit-related interlinkages of banks. As a result of our study, the dynamics underlying the meltdown of financial markets in 2008 becomes much better understandable.

  20. Thermal cascaded lattice Boltzmann method

    CERN Document Server

    Fei, Linlin

    2016-01-01

    In this paper, a thermal cascaded lattice Boltzmann method (TCLBM) is developed in combination with the double-distribution-function (DDF) approach. A density distribution function relaxed by the cascaded scheme is employed to solve the flow field, and a total energy distribution function relaxed by the BGK scheme is used to solve temperature field, where two distribution functions are coupled naturally. The forcing terms are incorporated by means of central moments, which is consistent with the previous force scheme [Premnath \\emph{et al.}, Phys. Rev. E \\textbf{80}, 036702 (2009)] but the derivation is more intelligible and the evolution process is simpler. In the method, the viscous heat dissipation and compression work are taken into account, the Prandtl number and specific-heat ratio are adjustable, the external force is considered directly without the Boussinesq assumption, and the low-Mach number compressible flows can also be simulated. The forcing scheme is tested by simulating a steady Taylor-Green f...

  1. Depression Screening

    Science.gov (United States)

    ... Depression Screening Substance Abuse Screening Alcohol Use Screening Depression Screening (PHQ-9) - Instructions The following questions are ... this tool, there is also text-only version . Depression Screening - Manual Instructions The following questions are a ...

  2. An update on the HIV treatment cascade in children and adolescents.

    Science.gov (United States)

    Bobat, Raziya; Archary, Moherndran; Lawler, Melissa

    2015-11-01

    To provide an update on the HIV treatment cascade in children and adolescents. We reviewed the literature on the steps in the cascade, for the period 2014-2015. There remains high attrition of children with regards to early testing and linking those patients who are positive to early treatment. Barriers to screening and testing in children and adolescents are multifactorial. Linkage to pre-antiretroviral therapy care and retention in care are the main steps at which attrition occurs. There are a number of new formulations available for use in adolescents and children which offer more options for antiretroviral therapy treatment. Adherence levels appear to be reasonable in Africa and Asia; however, achieving viral load suppression remains a challenge. We have a long way to go to achieve decreased attrition at each step of the cascade and retain patients in care. Recent improvements in each step of the cascade are bringing us closer to achieving treatment success.

  3. Lens Coupled Quantum Cascade Laser

    Science.gov (United States)

    Hu, Qing (Inventor); Lee, Alan Wei Min (Inventor)

    2013-01-01

    Terahertz quantum cascade (QC) devices are disclosed that can operate, e.g., in a range of about 1 THz to about 10 THz. In some embodiments, QC lasers are disclosed in which an optical element (e.g., a lens) is coupled to an output facet of the laser's active region to enhance coupling of the lasing radiation from the active region to an external environment. In other embodiments, terahertz amplifier and tunable terahertz QC lasers are disclosed.

  4. Turbulence: does energy cascade exist?

    CERN Document Server

    Josserand, Christophe; Lehner, Thierry; Pomeau, Yves

    2016-01-01

    To answer the question whether a cascade of energy exists or not in turbulence, we propose a set of correlation functions able to test if there is an irreversible transfert of energy, step by step, from large to small structures. These tests are applied to real Eulerian data of a turbulent velocity flow, taken in the wind grid tunnel of Modane, and also to a prototype model equation for wave turbulence. First we demonstrate the irreversible character of the flow by using multi-time correlation function at a given point of space. Moreover the unexpected behavior of the test function leads us to connect irreversibility and finite time singularities (intermittency). Secondly we show that turbulent cascade exists, and is a dynamical process, by using a test function depending on time and frequency. The cascade shows up only in the inertial domain where the kinetic energy is transferred more rapidly (on average) from the wavenumber $k_{1}$ to $k_{2}$ than from $k_{1}$ to $k'_{2}$ larger than $k_{2}$.

  5. Population screening and cascade testing for carriers of SMA

    NARCIS (Netherlands)

    Smith, Melanie; Calabro, Vanessa; Chong, Belinda; Gardiner, Nicole; Cowie, Shannon; du Sart, Desiree

    Spinal muscular atrophy (SMA) is one of the most common autosomal-recessive diseases, caused by absence of both copies of the survival motor neuron 1 (SMN1) gene. Identification of SMA carriers has important implications for individuals with a family history and the general population. SMA carriers

  6. Preimplantation genetic screening.

    Science.gov (United States)

    Harper, Joyce C

    2017-01-01

    Preimplantation genetic diagnosis was first successfully performed in 1989 as an alternative to prenatal diagnosis for couples at risk of transmitting a genetic or chromosomal abnormality, such as cystic fibrosis, to their child. From embryos generated in vitro, biopsied cells are genetically tested. From the mid-1990s, this technology has been employed as an embryo selection tool for patients undergoing in vitro fertilisation, screening as many chromosomes as possible, in the hope that selecting chromosomally normal embryos will lead to higher implantation and decreased miscarriage rates. This procedure, preimplantation genetic screening, was initially performed using fluorescent in situ hybridisation, but 11 randomised controlled trials of screening using this technique showed no improvement in in vitro fertilisation delivery rates. Progress in genetic testing has led to the introduction of array comparative genomic hybridisation, quantitative polymerase chain reaction, and next generation sequencing for preimplantation genetic screening, and three small randomised controlled trials of preimplantation genetic screening using these new techniques indicate a modest benefit. Other trials are still in progress but, regardless of their results, preimplantation genetic screening is now being offered globally. In the near future, it is likely that sequencing will be used to screen the full genetic code of the embryo.

  7. Development of electromagnetic cascades in the atmosphere including the Landau-Pomeranchuk-Migdal effect

    Science.gov (United States)

    Streitmatter, R. E.; Stephens, S. A.

    1985-01-01

    Numerical solutions have been obtained for the one-dimensional atmospheric electromagnetic cascade diffusion equations, including the Landau-Pomeranchuk-Migdal and screening effects. Spectra produced by primary gamma rays of various energies are given at a number of deths in the atmosphere.

  8. Skin sensitizers in cosmetics and beyond: potential multiple mechanisms of action and importance of T-cell assays for in vitro screening.

    Science.gov (United States)

    Vukmanović, Stanislav; Sadrieh, Nakissa

    2017-05-01

    Allergic contact dermatitis (ACD) is a delayed-type hypersensitivity (DTH) reaction induced by repeated contact with sensitizers. The ability of a chemical to act as a sensitizer has most frequently been tested in animals. As the use of animals for these purposes is gradually and globally being phased out, there is a need for reliable in vitro surrogate assays. Currently proposed in vitro assays are designed to test four key events of the adverse outcome pathway (AOP) involving covalent modification of self-proteins by sensitizers (haptenation) and presentation of new antigens (hapten/carrier complexes) to the immune system. There appears to be imperfect alignment of in vitro assays with clinical and/or animal data, suggesting possibly additional mechanisms of ACD development. Indeed, studies on allergies to small drugs, small chemical-induced HLA-peptide exchange for vaccination purposes and cosmetic ingredient-induced exposure of autoantigens suggest a possibility of DTH response promotion by hapten/carrier-independent mechanisms. Therefore, there is a need for additional appropriate in vitro assays, in order to achieve maximal concordance between clinical and/or animal data and in vitro assays. In this paper, we will review evidence supporting the idea of diverse mechanisms of ACD development. We will also discuss the impact of these multiple mechanisms, on the AOP and on the in vitro assays that should be used for allergen detection. We will propose alloreactivity-like reactions, aided by computer modeling and biochemical tests of compound-HLA binding, as additional tools for better prediction of DTH reactions, resulting from exposure to ingredients in cosmetic products. The combination of the proposed tests, along with the existing assays, should further enhance animal-free assessment of sensitizing potential of individual chemicals.

  9. Azobenzene-functionalized cascade molecules

    DEFF Research Database (Denmark)

    Archut, A.; Vogtle, F.; De Cola, L.

    1998-01-01

    Cascade molecules bearing up to 32 azobenzene groups in the periphery have been prepared from poly(propylene imine) dendrimers and N-hydroxysuccinimide esters. The dendritic azobenzene species show similar isomerization properties as the corresponding azobenzene monomers. The all-E azobenzene...... shows there is so far no effective steric constraint towards photoisomerism on increasing dimension (generation) of the dendrimer, The first attempts to use dendrimers for holography materials are described: It is shown that holographic gratings with diffraction efficiencies up to about 20 % can...

  10. Disaster Mythology and Availability Cascades

    Directory of Open Access Journals (Sweden)

    Lisa Grow Sun

    2013-04-01

    Full Text Available Sociological research conducted in the aftermath of natural disasters has uncovered a number of “disaster myths” – widely shared misconceptions about typical post-disaster human behavior. This paper discusses the possibility that perpetuation of disaster mythology reflects an “availability cascade,” defined in prior scholarship as a “self-reinforcing process of collective belief formation by which an expressed perception triggers a chain reaction that gives the perception increasing plausibility through its rising availability in public discourse.” (Kuran and Sunstein 1999. Framing the spread of disaster mythology as an availability cascade suggests that certain tools may be useful in halting the myths’ continued perpetuation. These tools include changing the legal and social incentives of so-called “availability entrepreneurs” – those principally responsible for beginning and perpetuating the cascade, as well as insulating decision-makers from political pressures generated by the availability cascade. This paper evaluates the potential effectiveness of these and other solutions for countering disaster mythology. Las investigaciones sociológicas realizadas tras los desastres naturales han hecho evidentes una serie de “mitos del desastre”, conceptos erróneos ampliamente compartidos sobre el comportamiento humano típico tras un desastre. Este artículo analiza la posibilidad de que la perpetuación de los mitos del desastre refleje una “cascada de disponibilidad”, definida en estudios anteriores como un “proceso de auto-refuerzo de la formación de una creencia colectiva, a través del que una percepción expresada produce una reacción en cadena que hace que la percepción sea cada vez más verosímil, a través de una mayor presencia en el discurso público” (Kuran y Sunstein 1999. Enmarcar la propagación de los mitos del desastre como una cascada de disponibilidad sugiere que ciertas herramientas pueden ser

  11. A Pax3/Dmrt2/Myf5 regulatory cascade functions at the onset of myogenesis.

    Directory of Open Access Journals (Sweden)

    Takahiko Sato

    2010-04-01

    Full Text Available All skeletal muscle progenitor cells in the body derive from the dermomyotome, the dorsal epithelial domain of developing somites. These multipotent stem cells express Pax3, and this expression is maintained in the myogenic lineage where Pax3 plays an important role. Identification of Pax3 targets is therefore important for understanding the mechanisms that underlie the onset of myogenesis. In a microarray screen of Pax3-GFP sorted cells, with analysis on Pax3 gain and loss of function genetic backgrounds, we identify Dmrt2, expressed in the dermomyotome, as a Pax3 target. In vitro gel shift analysis and chromatin immunoprecipitation with in vivo extracts show that Pax3 binds to a conserved 286 bp sequence, situated at -18 kb from Dmrt2. This sequence directs reporter transgene expression to the somite, and this is severely affected when the Pax3 site is mutated in the context of the locus. In Dmrt2 mutant embryos, somite maturation is perturbed and the skeletal muscle of the myotome is abnormal. We now report that the onset of myogenesis is also affected. This depends on activation, in the epaxial dermomyotome, of the myogenic determination gene, Myf5, through its early epaxial enhancer. This sequence contains sites that bind Dmrt2, which belongs to the DM class of DNA-binding proteins. Mutation of these sites compromises activity of the enhancer in transgenic embryos where the reporter transgene is under the control of the Myf5 epaxial enhancer. Transactivation of this site by Dmrt2 is demonstrated in vitro, and conditional overexpression of Dmrt2 in Pax3 expressing cells in the somite confirms the role of this factor in the activation of Myf5. These results reveal a novel genetic network, comprising a Pax3/Dmrt2/Myf5 regulatory cascade that operates in stem cells of the epaxial dermomyotome to initiate skeletal muscle formation.

  12. WHISTLER TURBULENCE FORWARD CASCADE VERSUS INVERSE CASCADE: THREE-DIMENSIONAL PARTICLE-IN-CELL SIMULATIONS

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Ouliang [Oracle Corporation, Redwood Shores, CA (United States); Gary, S. Peter [Space Science Institute, Boulder, CO (United States); Wang, Joseph, E-mail: ouliang@usc.edu, E-mail: pgary@lanl.gov, E-mail: josephjw@usc.edu [University of Southern California, Los Angeles, CA (United States)

    2015-02-20

    We present the results of the first fully three-dimensional particle-in-cell simulations of decaying whistler turbulence in a magnetized, homogeneous, collisionless plasma in which both forward cascades to shorter wavelengths, and inverse cascades to longer wavelengths are allowed to proceed. For the electron beta β {sub e} = 0.10 initial value considered here, the early-time rate of inverse cascade is very much smaller than the rate of forward cascade, so that at late times the fluctuation energy in the regime of the inverse cascade is much weaker than that in the forward cascade regime. Similarly, the wavevector anisotropy in the inverse cascade regime is much weaker than that in the forward cascade regime.

  13. Development and validation of a high-content screening in vitro micronucleus assay in CHO-k1 and HepG2 cells

    NARCIS (Netherlands)

    Westerink, W.M.; Schirris, T.J.J.; Horbach, G.J.; Schoonen, W.G.

    2011-01-01

    In the present study an automated image analysis assisted in vitro micronucleus assay was developed with the rodent cell line CHO-k1 and the human hepatoma cell line HepG2, which are both commonly used in regulatory genotoxicity assays. The HepG2 cell line was chosen because of the presence in these

  14. A workflow to investigate exposure and pharmacokinetic influences on high-throughput in vitro chemical screening based on adverse outcome pathways, OpenTox USA 2015 Poster

    Science.gov (United States)

    Adverse outcome pathways (AOP) link known population outcomes to a molecular initiating event (MIE) that can be quantified using high-throughput in vitro methods. Practical application of AOPs in chemical-specific risk assessment requires consideration of exposure and absorption,...

  15. Screening of bacterial isolates from various European soils for in vitro antagonistic activity towards Rhizoctonia solani and Fusarium oxysporum : Site-dependent composition and diversity revealed

    NARCIS (Netherlands)

    Adesina, Modupe F.; Lembke, Antje; Costa, Rodrigo; Speksnijder, Arien; Smalla, Kornelia

    2007-01-01

    A cultivation-based approach was used to determine the in vitro antagonistic potential of soil bacteria towards Rhizoctonia solani AG3 and Fusarium oxysporum f. sp. lini (Foln3). Four composite soil samples were collected from four agricultural sites with previous documentation of disease

  16. Screening of bacterial isolates from various European soils for in vitro antagonistic activity towards Rhizoctonia solani and Fusarium oxysporum: Site-dependent composition and diversity revealed

    NARCIS (Netherlands)

    Adesina, M.F.; Lembke, A.; Costa, R.; Speksnijder, A.G.C.L.; Smalla, K.

    2007-01-01

    A cultivation-based approach was used to determine the in vitro antagonistic potential of soil bacteria towards Rhizoctonia solani AG3 and Fusarium oxysporum f. sp. lini (Foln3). Four composite soil samples were collected from four agricultural sites with previous documentation of disease

  17. Contingency Analysis of Cascading Line Outage Events

    Energy Technology Data Exchange (ETDEWEB)

    Thomas L Baldwin; Magdy S Tawfik; Miles McQueen

    2011-03-01

    As the US power systems continue to increase in size and complexity, including the growth of smart grids, larger blackouts due to cascading outages become more likely. Grid congestion is often associated with a cascading collapse leading to a major blackout. Such a collapse is characterized by a self-sustaining sequence of line outages followed by a topology breakup of the network. This paper addresses the implementation and testing of a process for N-k contingency analysis and sequential cascading outage simulation in order to identify potential cascading modes. A modeling approach described in this paper offers a unique capability to identify initiating events that may lead to cascading outages. It predicts the development of cascading events by identifying and visualizing potential cascading tiers. The proposed approach was implemented using a 328-bus simplified SERC power system network. The results of the study indicate that initiating events and possible cascading chains may be identified, ranked and visualized. This approach may be used to improve the reliability of a transmission grid and reduce its vulnerability to cascading outages.

  18. Physics of interband cascade lasers

    Science.gov (United States)

    Vurgaftman, I.; Bewley, W. W.; Merritt, C. D.; Canedy, C. L.; Kim, C. S.; Abell, J.; Meyer, J. R.; Kim, M.

    2012-01-01

    The interband cascade laser (ICL) is a unique device concept that combines the effective parallel connection of its multiple-quantum-well active regions, interband active transitions, and internal generation of electrons and holes at a semimetallic interface within each stage of the device. The internal generation of carriers becomes effective under bias, and the role of electrical injection is to replenish the carriers consumed by recombination processes. Major strides have been made toward fundamentally understanding the rich and intricate ICL physics, which has in turn led to dramatic improvements in the device performance. In this article, we review the physical principles of the ICL operation and designs of the active region, electron and hole injectors, and optical waveguide. The results for state-of- the-art ICLs spanning the 3-6 μm wavelength range are also briefly reviewed. The cw threshold input powers at room temperature are more than an order of magnitude lower than those for quantum cascade lasers throughout the mid-IR spectral range. This will lengthen battery lifetimes and greatly relax packaging and size/weight requirements for fielded sensing systems.

  19. Lifespans of Cascade Arc volcanoes

    Science.gov (United States)

    Calvert, A. T.

    2015-12-01

    Compiled argon ages reveal inception, eruptive episodes, ages, and durations of Cascade stratovolcanoes and their ancestral predecessors. Geologic mapping and geochronology show that most Cascade volcanoes grew episodically on multiple scales with periods of elevated behavior lasting hundreds of years to ca. 100 kyr. Notable examples include the paleomag-constrained, few-hundred-year-long building of the entire 15-20 km3 Shastina edifice at Mt. Shasta, the 100 kyr-long episode that produced half of Mt. Rainier's output, and the 30 kyr-long episode responsible for all of South and Middle Sister. Despite significant differences in timing and rates of construction, total durations of active and ancestral volcanoes at discrete central-vent locations are similar. Glacier Peak, Mt. Rainier, Mt. Adams, Mt. Hood, and Mt. Mazama all have inception ages of 400-600 ka. Mt. St. Helens, Mt. Jefferson, Newberry Volcano, Mt. Shasta and Lassen Domefield have more recent inception ages of 200-300 ka. Only the Sisters cluster and Mt. Baker have established eruptive histories spanning less than 50 kyr. Ancestral volcanoes centered 5-20 km from active stratocones appear to have similar total durations (200-600 kyr), but are less well exposed and dated. The underlying mechanisms governing volcano lifecycles are cryptic, presumably involving tectonic and plumbing changes and perhaps circulation cycles in the mantle wedge, but are remarkably consistent along the arc.

  20. A Semisupervised Cascade Classification Algorithm

    Directory of Open Access Journals (Sweden)

    Stamatis Karlos

    2016-01-01

    Full Text Available Classification is one of the most important tasks of data mining techniques, which have been adopted by several modern applications. The shortage of enough labeled data in the majority of these applications has shifted the interest towards using semisupervised methods. Under such schemes, the use of collected unlabeled data combined with a clearly smaller set of labeled examples leads to similar or even better classification accuracy against supervised algorithms, which use labeled examples exclusively during the training phase. A novel approach for increasing semisupervised classification using Cascade Classifier technique is presented in this paper. The main characteristic of Cascade Classifier strategy is the use of a base classifier for increasing the feature space by adding either the predicted class or the probability class distribution of the initial data. The classifier of the second level is supplied with the new dataset and extracts the decision for each instance. In this work, a self-trained NB∇C4.5 classifier algorithm is presented, which combines the characteristics of Naive Bayes as a base classifier and the speed of C4.5 for final classification. We performed an in-depth comparison with other well-known semisupervised classification methods on standard benchmark datasets and we finally reached to the point that the presented technique has better accuracy in most cases.

  1. Extraction and Bioactivity Analysis of Major Flavones Compounds from Scutellaria baicalensis Using In Vitro Assay and Online Screening HPLC-ABTS System

    Directory of Open Access Journals (Sweden)

    Kwang Jin Lee

    2014-01-01

    Full Text Available The extraction efficiency of a number of solvent compositions for the improvement of bioactive compounds yield from S. baicalensis has been investigated. Also, free radical scavengers in the glycoside baicalin (BG, wogonoside (WG, aglycon baicalein (B, and wogonin (W compounds of S. baicalensis were screened, identified, and quantified using coupled offline ABTS and online screening HPLC-ABTS assay. Increasing ethanol content fractions resulted in decreased extract yield of bioactive compounds. In this case, the best yield of 37.01 mg/g in BG, WG, B, and W compounds was obtained by a dipping method with an extraction time of 4 h. In addition, the yield (43.05% and IC50 (34.04 μg/mL determined through ABTS assay of the 60% aqueous ethanol extract were the most satisfactory of all solvent solutions tested. This result shows that an online screening HPLC-ABTS assay can be a powerful technique for the rapid characterization of bioactivity compounds in plant extracts. Moreover, their anti-inflammatory activities were evaluated via analyzed inhibitory effect on NO and inflammatory cytokine production. Furthermore, WG and W exhibited the strong inhibitory effects on inflammatory mediator production including NO, IL-6, and IL-1β in LPS-stimulated RAW 264.7 macrophages.

  2. Power quality enhancement using cascaded multilevel inverter ...

    African Journals Online (AJOL)

    This paper investigates mitigation of current harmonics using different configuration of cascaded multilevel inverter based shunt hybrid active power filter (SHAPF) and to improve power quality of the system. The main objective of this paper is to develop and analyze the compensation characteristics of cascaded multilevel ...

  3. Cascading disaster models in postburn flash flood

    Science.gov (United States)

    Fred May

    2007-01-01

    A useful method of modeling threats from hazards and documenting their disaster causation sequences is called “cascading threat modeling.” This type of modeling enables emergency planners to address hazard and risk assessments systematically. This paper describes a cascading threat modeling and analysis process. Wildfire and an associated postburn flash flood disaster...

  4. Cascade Harvest’ red raspberry

    Science.gov (United States)

    Cascade Harvest’ is a new floricane fruiting raspberry cultivar (Rubus idaeus L.) jointly released by Washington State University (WSU), Oregon State University (OSU) and the U.S. Department of Agriculture (USDA). ‘Cascade Harvest’ produces a high yield of large, firm fruit suited to machine harves...

  5. Suppressing cascades of load in interdependent networks

    CERN Document Server

    Brummitt, Charles D; Leicht, E A

    2011-01-01

    Understanding how interdependence among systems affects cascading behaviors is increasingly important across many fields of science and engineering. Inspired by cascades of load shedding in coupled electric grids and other infrastructure, we study the Bak-Tang-Wiesenfeld sandpile model on modular random graphs and on graphs based on actual, interdependent power grids. Starting from two isolated networks, adding some connectivity between them is beneficial, for it suppresses the largest cascades in each system. Too much interconnectivity, however, becomes detrimental for two reasons. First, interconnections open pathways for neighboring networks to inflict large cascades. Second, as in real infrastructure, new interconnections increase capacity and total possible load, which fuels even larger cascades. Using a multi-type branching process and simulations we show these effects and estimate the optimal level of interconnectivity that balances their tradeoffs. Such equilibria could allow, for example, power grid ...

  6. MAPK cascades in guard cell signal transduction

    Directory of Open Access Journals (Sweden)

    Yuree eLee

    2016-02-01

    Full Text Available Guard cells form stomata on the epidermis and continuously respond to endogenous and environmental stimuli to fine-tune the gas exchange and transpirational water loss, processes which involve mitogen-activated protein kinase (MAPK cascades. MAPKs form three-tiered kinase cascades with MAPK kinases and MAPK kinase kinases, by which signals are transduced to the target proteins. MAPK cascade genes are highly conserved in all eukaryotes, and they play crucial roles in myriad developmental and physiological processes. MAPK cascades function during biotic and abiotic stress responses by linking extracellular signals received by receptors to cytosolic events and gene expression. In this review, we highlight recent findings and insights into MAPK-mediated guard cell signaling, including the specificity of MAPK cascades and the remaining questions.

  7. Experimental study of flow through compressor Cascade

    Directory of Open Access Journals (Sweden)

    Satyam Panchal

    2017-09-01

    Full Text Available The objective of this research work is to study the behaviour of flow at the inlet, within the blade passage and at the exit of a compressor cascade. For this purpose, a cascade with six numbers of aerofoil blades was designed and constructed. The cascade was fitted on the cascade test tunnel. Out of six blades two were instrumented for measuring the pressure distribution on the pressure and suction surface. The blades had a parabolic camber line, with a maximum camber position at 40% of the chord from the leading edge of the blade. The profile of the blade was C4, height of the blade was 160 mm, chord length was 80 mm, camber angle was 45° and stagger angle was 30°. Similarly, the length of the cascade was 300 mm, span was 160 mm, pitch was 60 mm, the actual chord of the cascade was 80 mm, the axial chord of the cascade was 70 mm, the stagger angle of the cascade was 30° and the pitch-chord ratio was 0.75. The data was taken and analyzed at −500% of the axial chord before the cascade, −25% of the axial chord before the leading edge, 25%, 50%, 75% and 150% of the axial chord from the leading edge of the blade. The readings were taken from the cascade wall to the mid span position along the pitch wise direction. The angle of incidence was also changed during the experiment and varied from i=−50°, −30°, −10° to 5°.

  8. Screening for in vitro shoot-forming capacity of seedling explants in bell pepper (Capsicum annuum L.) genotypes and efficient plant regeneration using thidiazuron.

    Science.gov (United States)

    Szász, A; Nervo, G; Fári, M

    1995-07-01

    In vitro shoot regeneration ability of 17 (7 Italian and 10 Hungarian) bell pepper genotypes was investigated using excised cotyledons and rooted hypocotyls as explants. Most of the Italian genotypes and two of the Hungarian genotypes responded well, producing shoots from rooted hypocotyls. Only two genotypes (one Italian and one Hungarian) gave a weak response using cotyledons. For direct shoot induction in these explants, in addition to the methods cited in the relevant papers, a new method was applied using thidiazuron as a cytokinin. Shoots were successfully regenerated from cotyledons of two Italian and two Hungarian genotypes using thidiazuron which were considered to be non responsive to the usual methods.

  9. Successful pregnancy outcome after in vitro fertilisation following Pre-implantation Genetic Diagnosis/Polymerase Chain Reaction screening for single gene disorder (sickle cell anaemia) before embryo transfer: The clinical experience of an in vitro fertilisation clinic in Nigeria.

    Science.gov (United States)

    Okeke, Chizara; Ailoje-Ibru, Kemi; Olukoya, Kemi; Ogbeche, Rose; Adewusi, Abiola; Iloabachie, Ebele; Ashiru, Oladapo

    2014-01-01

    A couple, both carriers of the sickle cell anaemia trait (Genotype HbAS) with an offspring already affected with the genetic disease underwent a Pre-implantation Genetic Diagnosis/Polymerase Chain Reaction screening of biopsied blastomeres. DNA analysis of single blastomeres was carried out to find out indicated a viable intra-uterine pregnancy with embryos which carried the sickle cell mutation, which resulted in a livebirth (HbAS). PGD/PCR in combination with IVF appears to be the most suitable treatment plan for patients who are at a higher risk of reproducing offspring affected with inheritable genetic diseases.

  10. Successful pregnancy outcome after in vitro fertilisation following Pre-implantation Genetic Diagnosis/Polymerase Chain Reaction screening for single gene disorder (sickle cell anaemia) before embryo transfer: The clinical experience of an in vitro fertilisation clinic in Nigeria

    OpenAIRE

    Chizara Okeke; Kemi Ailoje-Ibru; Kemi Olukoya; Rose Ogbeche; Abiola Adewusi; Ebele Iloabachie; Oladapo Ashiru

    2014-01-01

    A couple, both carriers of the sickle cell anaemia trait (Genotype HbAS) with an offspring already affected with the genetic disease underwent a Pre-implantation Genetic Diagnosis/Polymerase Chain Reaction screening of biopsied blastomeres. DNA analysis of single blastomeres was carried out to find out indicated a viable intra-uterine pregnancy with embryos which carried the sickle cell mutation, which resulted in a livebirth (HbAS). PGD/PCR in combination with IVF appears to be the most suit...

  11. Drug Repositioning for Preeclampsia Therapeutics by In Vitro Screening: Phosphodiesterase-5 Inhibitor Vardenafil Restores Endothelial Dysfunction via Induction of Placental Growth Factor.

    Science.gov (United States)

    Kakigano, Aiko; Tomimatsu, Takuji; Mimura, Kazuya; Kanayama, Tomoko; Fujita, Satoko; Minato, Kenji; Kumasawa, Keiichi; Taniguchi, Yukiko; Kanagawa, Takeshi; Endo, Masayuki; Ishihara, Tomoaki; Namba, Takushi; Mizushima, Tohru; Kimura, Tadashi

    2015-10-01

    We screened a library of 528 approved drugs to identify candidate compounds with therapeutic potential as preeclampsia treatments via their proangiogenic properties. Using human umbilical vein endothelial cells (HUVECs), we assessed whether the screened drugs induced placental growth factor (PIGF) and restored damaged endothelial cell function. Enzyme-linked immunosorbent assays (ELISAs) were carried out to measure levels of PlGF in conditioned media treated with each drug (100 µmol/L) in the drug library. Tube formation assays were performed using HUVECs to evaluate the angiogenic effects of drugs that induced PlGF. We also performed ELISA, quantitative reverse transcription polymerase chain reaction, and tube formation assays after treatment with a range of concentrations of the candidate drug. Of the drugs that induced PlGF, vardenafil was the only compound that significantly facilitated tube formation in comparison with the control cells (P Treatment with vardenafil at concentrations of 50, 100, and 250 µmol/L increased expression of PlGF in a dose-dependent manner. Vardenafil (250 µmol/L) significantly improved tube formation which was inhibited in the presence of soluble fms-like tyrosine kinase 1 (100 ng/mL) and/or soluble endoglin (100 ng/mL). Production of PlGF from HUVECs in the presence of sera derived from patients with preeclampsia was significantly elevated by administration of vardenafil (250 µmol/L). By assessing drug repositioning through screening a library of approved drugs, we identified vardenafil as a potential protective agent against preeclampsia. The therapeutic mechanism of vardenafil may involve inhibition of the systemic maternal antiangiogenic state that leads to preeclampsia, in addition to its vasodilating effect. As concentrations used are high and unlikely to be useful clinically, further work is needed before testing it in humans. © The Author(s) 2015.

  12. Product lifecycle approach to cascade impaction measurements.

    Science.gov (United States)

    Tougas, Terrence P; Christopher, Dave; Mitchell, Jolyon; Lyapustina, Svetlana; Van Oort, Michiel; Bauer, Richard; Glaab, Volker

    2011-03-01

    Over the lifecycle of an orally inhaled product (OIP), multi-stage cascade impactor (CI) measurements are used for different purposes and to address different questions. Full-resolution CIs can provide important information during product development and are widely used but are time- and resource-intensive, highly variable, and suboptimal for OIP quality control (QC) testing. By contrast, Efficient Data Analysis (EDA) combined with Abbreviated Impactor Measurement (AIM) systems pertinent either for QC and-possibly-for adult Human Respiratory Tract (pHRT) has been introduced for OIP performance assessment during and post-development. This article summarizes available evidence and discusses a strategy for using either abbreviated or full-resolution CI systems depending on the purpose of the measurement, such that adequate, accurate, and efficient testing of aerodynamic particle size distribution (APSD) of OIPs can be achieved throughout the lifecycle of a product. Under these proposals, a comprehensive testing program should initially be conducted by full-resolution CI in OIP development to ascertain the product's APSD. Subsequently, correlations should be established from the selected AIM CIs to the corresponding full-resolution system, ideally developing specifications common to both techniques. In the commercial phase, it should be possible to release product using AIM/EDA, keeping the full-resolution CI for investigations, change control, and trouble-shooting, thus optimizing resources for APSD characterization throughout the product lifecycle. If an in vitro-in vivo relationship is established and clinically relevant sizes are known, an AIM-pHRT could serve as a quick indicator that clinically relevant fractions have not changed and also, in the management of post-approval changes. © 2011 American Association of Pharmaceutical Scientists

  13. Screening of the anthocyanin profile and in vitro pancreatic lipase inhibition by anthocyanin-containing extracts of fruits, vegetables, legumes and cereals.

    Science.gov (United States)

    Fabroni, Simona; Ballistreri, Gabriele; Amenta, Margherita; Romeo, Flora V; Rapisarda, Paolo

    2016-11-01

    The phytotherapic treatment of overweight and/or moderate obesity is growing widely, thus there is a great interest towards the phenolic compounds of fruits and vegetables which may inhibit pancreatic lipase enzyme. In this study, we report the chemical composition and in vitro pancreatic lipase inhibitory activity of 13 freeze-dried anthocyanin-containing extracts of different Mediterranean plants: fruits (blood orange, pomegranate, blackberry, mulberry and sumac), citrus by-products (blood orange peel), citrus vegetative tissues (young lemon shoots); vegetables (red cabbage and violet cauliflower), legume seeds (black bean), cereals (black rice), and cereal processing by-products (black rice hull). Total phenols and anthocyanins were determined. Individual anthocyanins were identified by UHPLC-PDA-ESI/MSn . Results revealed a wide variation in the distribution of anthocyanin compounds. Blood orange and pomegranate juice extracts had the highest total anthocyanin content and exhibited the strongest inhibition of pancreatic lipase in vitro. Inhibitory activity was positively correlated with anthocyanin content. In appropriate formulations, anthocyanin-containing extracts could find a use as anti-obesity agents. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  14. Method for screening sunscreen cream formulations by determination of in vitro SPF and PA values using UV transmission spectroscopy and texture profile analysis.

    Science.gov (United States)

    Khunkitti, Watcharee; Satthanakul, Panitta; Waranuch, Neti; Pitaksuteepong, Tasana; Kitikhun, Pichet

    2014-01-01

    Formulation of sunscreen products to obtain high values of sun protection factor (SPF) and protection from ultraviolet A (PA) is challenging work for cosmetic chemists. This study aimed to study factors affecting SPF and PA values using ultraviolet transmission spectroscopy as well as texture profiles of sunscreen formulations using 23 factorial designs. Results demonstrate that the correlation coefficient between the labeled SPF values of counter-brand sunscreen products and the in vitro SPF values was 0.901. In vitro SPF determination showed that the combination effect of phase volume ratio (PVR) and xanthan gum caused a significant increase to the SPF values of the formulations, whereas the interaction effect between PVR and stearic acid significantly decreased the SPF value. In addition, there was the interaction effect between xanthan gum and stearic acid leading to significant reduction of hardness, compressibility, and pH, but significantly increasing the adhesiveness. All tested factors did not significantly affect the cohesiveness of tested formulations. In conclusion, apart from sunscreen agents, the other ingredients also affected the SPF and PA values. The calculated SPF values range from 21 to 60. However, a selected formulation needs to be confirmed by the standard method of testing. In addition, the physical, chemical, and biological stability; shelf life; and sensory evaluation of all formulations need to be evaluated.

  15. Isolation and Bioactivity Analysis of Ethyl Acetate Extract from Acer tegmentosum Using In Vitro Assay and On-Line Screening HPLC-ABTS+ System

    Directory of Open Access Journals (Sweden)

    Kwang Jin Lee

    2014-01-01

    Full Text Available The Acer tegmentosum (3 kg was extracted using hot water, and the freeze-dried extract powder was partitioned successively using dichloromethane (DCM, ethyl acetate (EA, butyl alcohol (n-BuOH, and water. From the EA extract fraction (1.24 g, five phenolic compounds were isolated by the silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatography. Based on spectroscopic methods such as 1H-NMR, 13C-NMR, and LC/MS the chemical structures of the compounds were confirmed as feniculin (1, avicularin (2, (+-catechin (3, (−-epicatechin (4, and 6′-O-galloyl salidroside (5. Moreover, a rapid on-line screening HPLC-ABTS+ system for individual bioactivity of the EA-soluble fraction (five phenolic compounds was developed. The results indicated that compounds 1 and 2 were first isolated from the A. tegmentosum. The anti-inflammatory activities and on-line screening HPLC-ABTS+ assay method of these compounds in LPS-stimulated murine macrophages were rapid and efficient for the investigation of bioactivity of A. tegmentosum.

  16. MAPK cascade signalling networks in plant defence.

    Science.gov (United States)

    Pitzschke, Andrea; Schikora, Adam; Hirt, Heribert

    2009-08-01

    The sensing of stress signals and their transduction into appropriate responses is crucial for the adaptation and survival of plants. Kinase cascades of the mitogen-activated protein kinase (MAPK) class play a remarkably important role in plant signalling of a variety of abiotic and biotic stresses. MAPK cascade-mediated signalling is an essential step in the establishment of resistance to pathogens. Here, we describe the most recent insights into MAPK-mediated pathogen defence response regulation with a particular focus on the cascades involving MPK3, MPK4 and MPK6. We also discuss the strategies developed by plant pathogens to circumvent, inactivate or even 'hijack' MAPK-mediated defence responses.

  17. Monitoring drug self-aggregation and potential for promiscuity in off-target in vitro pharmacology screens by a practical NMR strategy.

    Science.gov (United States)

    LaPlante, Steven R; Aubry, Norman; Bolger, Gordon; Bonneau, Pierre; Carson, Rebekah; Coulombe, René; Sturino, Claudio; Beaulieu, Pierre L

    2013-09-12

    A simple NMR assay was applied to monitor the tendency of compounds to self-aggregate in aqueous media. The observation of unusual spectral trends as a function of compound concentration appears to be signatory of the formation of self-assemblies. (1)H NMR resonances of aggregating compounds were sensitive to the presence of a range of molecular assemblies in solution including large molecular-size entities, smaller multimers, and mixtures of assembled species. The direct observation of aggregates via unusual NMR spectra also correlated with promiscuous behavior of molecules in off-target in vitro pharmacology assays. This empirical assay can have utility for predicting compound promiscuity and should complement predictive methods that principally rely on the computing of descriptors such as lipophilicity (cLogP) and topological surface area (TPSA). This assay should serve as a practical tool for medicinal chemists to monitor compound attributes in aqueous solution and various pharmacologically relevant media, as demonstrated herein.

  18. Screening of in vitro cytotoxicity, antioxidant potential and bioactivity of nano- and micro-ZrO2 and -TiO2 particles.

    Science.gov (United States)

    Karunakaran, Gopalu; Suriyaprabha, Rangaraj; Manivasakan, Palanisamy; Yuvakkumar, Rathinam; Rajendran, Venkatachalam; Kannan, Narayanasamy

    2013-07-01

    Nanometal oxides are used in tissue engineering and implants. The increased use of nanoparticles suggests the need to study their adverse effects on biological systems. The present investigation explores in vitro cytotoxicity, antioxidant potential, and bioactivity of nano- and micro-particles such as zirconia (ZrO2) and titania (TiO2) on biological systems such as National Institute of Health (NIH) 3T3 mouse embryonic fibroblasts cell line, di(phenyl)-(2,4,6-trinitrophenyl) iminoazanium (DPPH) and simulated body fluid (SBF). The cell line viability % indicated that nano ZrO2 and TiO2 were less toxic than microparticles up to 200µgml(-1). DPPH assay revealed that the free radical scavenging potential of tested particles were higher for nano ZrO2 (76.9%) and nano TiO2 (73.3%) at 100mg than that for micron size particles. Calcium deposition percentage of micro- and nano-ZrO2 particles, after SBF study, showed 0.066% and 0.094% respectively, whereas for micro- and nano-TiO2, it was 0.251% and 0.615% respectively. FTIR results showed a good bioactivity through hydroxyapatite formation. The present investigation clearly shows that nanoparticles possess good antioxidant potential and better biocompatibility under in vitro conditions which are dose and size dependent. Hence, cytotoxicity itself is not promising evaluation method for toxicity rather than particles individual characterisation using antioxidant and bioactivity analysis. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. A Sensitive in Vitro High-Throughput Screen To Identify Pan-filoviral Replication Inhibitors Targeting the VP35–NP Interface

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Gai; Nash, Peter J.; Johnson, Britney; Pietzsch, Colette; Ilagan, Ma. Xenia G.; Bukreyev, Alexander; Basler, Christopher F.; Bowlin, Terry L.; Moir, Donald T.; Leung, Daisy W.; Amarasinghe, Gaya K. (WU-MED); (GSU); (Texas-MED); (Microbiotix)

    2017-01-24

    The 2014 Ebola outbreak in West Africa, the largest outbreak on record, highlighted the need for novel approaches to therapeutics targeting Ebola virus (EBOV). Within the EBOV replication complex, the interaction between polymerase cofactor, viral protein 35 (VP35), and nucleoprotein (NP) is critical for viral RNA synthesis. We recently identified a peptide at the N-terminus of VP35 (termed NPBP) that is sufficient for interaction with NP and suppresses EBOV replication, suggesting that the NPBP binding pocket can serve as a potential drug target. Here we describe the development and validation of a sensitive high-throughput screen (HTS) using a fluorescence polarization assay. Initial hits from this HTS include the FDA-approved compound tolcapone, whose potency against EBOV infection was validated in a nonfluorescent secondary assay. High conservation of the NP–VP35 interface among filoviruses suggests that this assay has the capacity to identify pan-filoviral inhibitors for development as antivirals.

  20. In vitro screening for anti-cholinesterase and antioxidant activity of methanolic extracts of ayurvedic medicinal plants used for cognitive disorders.

    Directory of Open Access Journals (Sweden)

    Maya Mathew

    Full Text Available Inhibition of Acetylcholinesterase (AChE is still considered as the main therapeutic strategy against Alzheimer's disease (AD. Many plant derived phytochemicals have shown AChE inhibitory activity in addition to the currently approved drugs for AD. In the present study, methanolic extracts of 20 plants used in Indian Ayurvedic system of medicine for improving cognitive function were screened for acetylcholinesterase inhibitory activity by Ellman's microplate colorimetric method. Out of 20 extracts, Emblica officinalis, Nardostachys jatamansi, Nelumbo nucifera, Punica granatum and Raulfia Serpentina showed IC50 values <100 µg/ml for acetylcholinesterase inhibitory activity. Antioxidant activities of these plants were assessed by DPPH scavenging assay. Among the extracts used, antioxidant activity was highest for Terminalia chebula and Emblica officinalis with IC50 values <10 µg/ml. Considering the complex multifactorial etiology of AD, these plant extracts will be safer and better candidates for the future disease modifying therapies against this devastating disease.

  1. Mount Rainier active cascade volcano

    Science.gov (United States)

    1994-01-01

    Mount Rainier is one of about two dozen active or recently active volcanoes in the Cascade Range, an arc of volcanoes in the northwestern United States and Canada. The volcano is located about 35 kilometers southeast of the Seattle-Tacoma metropolitan area, which has a population of more than 2.5 million. This metropolitan area is the high technology industrial center of the Pacific Northwest and one of the commercial aircraft manufacturing centers of the United States. The rivers draining the volcano empty into Puget Sound, which has two major shipping ports, and into the Columbia River, a major shipping lane and home to approximately a million people in southwestern Washington and northwestern Oregon. Mount Rainier is an active volcano. It last erupted approximately 150 years ago, and numerous large floods and debris flows have been generated on its slopes during this century. More than 100,000 people live on the extensive mudflow deposits that have filled the rivers and valleys draining the volcano during the past 10,000 years. A major volcanic eruption or debris flow could kill thousands of residents and cripple the economy of the Pacific Northwest. Despite the potential for such danger, Mount Rainier has received little study. Most of the geologic work on Mount Rainier was done more than two decades ago. Fundamental topics such as the development, history, and stability of the volcano are poorly understood.

  2. Studies of Cream Seeded Carioca Beans (Phaseolus vulgaris L.) from a Rwandan Efficacy Trial: In Vitro and In Vivo Screening Tools Reflect Human Studies and Predict Beneficial Results from Iron Biofortified Beans.

    Science.gov (United States)

    Tako, Elad; Reed, Spenser; Anandaraman, Amrutha; Beebe, Steve E; Hart, Jonathan J; Glahn, Raymond P

    2015-01-01

    targeting of such compounds during the breeding process may yield improved dietary Fe-bioavailability. Our findings are in agreement with the human efficacy trial that demonstrated that the biofortified carioca beans improved the Fe-status of Rwandan women. We suggest the utilization of these in vitro and in vivo screening tools to guide studies aimed to develop and evaluate biofortified staple food crops. This approach has the potential to more effectively utilize research funds and provides a means to monitor the nutritional quality of the Fe-biofortified crops once released to farmers.

  3. Studies of Cream Seeded Carioca Beans (Phaseolus vulgaris L. from a Rwandan Efficacy Trial: In Vitro and In Vivo Screening Tools Reflect Human Studies and Predict Beneficial Results from Iron Biofortified Beans.

    Directory of Open Access Journals (Sweden)

    Elad Tako

    , specific targeting of such compounds during the breeding process may yield improved dietary Fe-bioavailability. Our findings are in agreement with the human efficacy trial that demonstrated that the biofortified carioca beans improved the Fe-status of Rwandan women. We suggest the utilization of these in vitro and in vivo screening tools to guide studies aimed to develop and evaluate biofortified staple food crops. This approach has the potential to more effectively utilize research funds and provides a means to monitor the nutritional quality of the Fe-biofortified crops once released to farmers.

  4. Synthesis, in-vitro antimicrobial and antitubercular screening of Schiff bases of 3-amino-1-phenyl-4- [2-(4-phenyl-1,3-thiazol-2-yl hydrazin-1-ylidene]-4,5-dihydro-1H-pyrazol-5-one

    Directory of Open Access Journals (Sweden)

    K K Sivakumar

    2013-01-01

    Full Text Available Purpose: Synthesis and antimicrobial activity of some Schiff bases of 3-amino-1-phenyl-4- [2-(4-phenyl-1,3-thiazol-2-yl hydrazin-1-ylidene]-4,5-dihydro-1H-pyrazol-5-ones (TZP4a-l are described. Materials and Methods: Structures of the synthesized compounds were confirmed using infrared, 1 H nuclear magnetic resonance, and mass spectral data. Synthesized compounds were tested in-vitro against four Gram-positive and four Gram-negative bacterial strains, three fungal strains and two mycobacterial strains. Title compounds were screened its in-vitro cytotoxicity (IC 50 by 3-(4,5-dimethythiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT assay using mouse embryonic fibroblasts cell line (NIH 3T3. Results and Discussion: Compounds TZP4 g and TZP4 h were found to be significant activity against Bacillus substilis (bacteria and Aspergillus niger (fungi. In-vitro anti-tuberculosis (TB activity of compound TZP4g showed appreciable antitubercular activity against Mycobacterium tuberculosis H37Rv strain (minimum inhibitory concentration [MIC] =0.6.48 × 10−3 μM/mL which was 1.69 and 3.91 times more active than the standard drug, pyrazinamide (25.38 × 10−3 μM/mL and streptomycin (MIC = 11.01 × 10−3 μM/mL, respectively. Their in-vitro cytotoxicity (IC 50 was determined to establish a selectivity index (SI (SI = IC 50 /MIC. Compounds TZP4 c, TZP4 g, and TZP4 h have SI 82.85, 168.88, and 199.07, respectively. Conclusion: All the title compounds had mild toxicity on the mouse embryonic fibroblasts NIH 3T3 cells (IC 50 ≥ 100 μM. In comparison to the results of toxicity and antimycobacterial activity tests, it was observed that the activity of the compounds is not due to general toxicity effect; however, their antimycobacterial activity can be possibly because of their selective antimycobacterial effect. We concluded from our investigations that TZP4 c, TZP4 g, and TZP4 h may be considered promising for the development of new anti-TB agents.

  5. Resilience and Regime Shifts: Assessing Cascading Effects

    National Research Council Canada - National Science Library

    Kinzig, Ann P; Ryan, Paul; Etienne, Michel; Allison, Helen; Elmqvist, Thomas; Walker, Brian H

    2006-01-01

    .... "Cascading thresholds," i.e., the tendency of the crossing of one threshold to induce the crossing of other thresholds, often lead to very resilient, although often less desirable, alternative states.

  6. Substrate-emitting ring interband cascade lasers

    Science.gov (United States)

    Holzbauer, Martin; Szedlak, Rolf; Detz, Hermann; Weih, Robert; Höfling, Sven; Schrenk, Werner; Koeth, Johannes; Strasser, Gottfried

    2017-10-01

    We demonstrate interband cascade lasers fabricated into ring-shaped cavities with vertical light emission through the substrate at a wavelength of λ ≈ 3.7 μm. The out-coupling mechanism is based on a metallized second-order distributed feedback grating. At room-temperature, a pulsed threshold current-density of 0.75 kA/cm2 and a temperature-tuning rate of 0.3 nm/°C are measured. In contrast to the azimuthal polarization of ring quantum cascade lasers, we observe a radial polarization of the projected nearfield of ring interband cascade lasers. These findings underline the fundamental physical difference between light generation in interband and intersubband cascade lasers, offering new perspectives for device integration.

  7. In-vitro screening of acetylcholinesterase inhibitory activity of extracts from Palestinian indigenous flora in relation to the treatment of Alzheimer’s disease

    Directory of Open Access Journals (Sweden)

    Mohammed Saleem Ali-Shtayeh

    2014-09-01

    Full Text Available Background: Cholinesterase inhibitory therapy serves as a strategy for the treatment of Alzheimer’s disease (AD. Several acetylcholinesterase inhibitors (AChEIs are used for the symptomatic treatment of AD. These compounds have been reported to have adverse effects, including gastrointestinal disturbances. This study was therefore partly aimed at investigating in vitro possible AChEIs in herbal medicines traditionally used in Palestine to treat cognitive disorders, and to point out the role of these plants as potential sources for development of newly potent and safe natural therapeutic agents of AD. Assay of AChE activity plays an important role in vitro characterization of drugs including potential treatments for AD. The most widely used method, is based on Ellman’s method. The reactant used in this method shows chemical reactivity with oxime antidots and thiol leading to false positive reactions. A new alternative assay could be of high interest. Methods: The effect on AChE activity of 92 extracts of 47 medicinal plants were evaluated using a new micro-well plate AChE activity (NA-FB and Ellman’s assays. In addition, antioxidant activity using DPPH was determined. Results: The main advantages of the new method (NA-FB is that the colorimetric change is better observable visually allowing spectrophotometric as well as colorimetric assay, and does not show any chemical reactivity with thiol. 67.4% and 37% of extracts inhibited AChE by >50% using the NA-FB and Ellman’s assays, respectively. Using NA-FB assay, 84 extracts interacted reversibly with the enzyme, of which Mentha spicata (94.8%, Foeniculum vulgare (89.81, and Oxalis pes-caprae (89.21 were most potent, and 8 showed irreversible inhibition of which leaves of Lupinus pilosus (92.02% were most active. Antioxidant activity was demonstrated by 73 extracts Majorana syriaca (IC50 0.21mg/ml, and Rosmarinus officinalis (0.38 were the most active. Conclusions: NA-FB assay has shown to be

  8. Compression limits in cascaded quadratic soliton compression

    DEFF Research Database (Denmark)

    Bache, Morten; Bang, Ole; Krolikowski, Wieslaw

    2008-01-01

    Cascaded quadratic soliton compressors generate under optimal conditions few-cycle pulses. Using theory and numerical simulations in a nonlinear crystal suitable for high-energy pulse compression, we address the limits to the compression quality and efficiency.......Cascaded quadratic soliton compressors generate under optimal conditions few-cycle pulses. Using theory and numerical simulations in a nonlinear crystal suitable for high-energy pulse compression, we address the limits to the compression quality and efficiency....

  9. Cascade of links in complex networks

    Energy Technology Data Exchange (ETDEWEB)

    Feng, Yeqian; Sun, Bihui [Department of Management Science, School of Government, Beijing Normal University, 100875 Beijing (China); Zeng, An, E-mail: anzeng@bnu.edu.cn [School of Systems Science, Beijing Normal University, 100875 Beijing (China)

    2017-01-30

    Cascading failure is an important process which has been widely used to model catastrophic events such as blackouts and financial crisis in real systems. However, so far most of the studies in the literature focus on the cascading process on nodes, leaving the possibility of link cascade overlooked. In many real cases, the catastrophic events are actually formed by the successive disappearance of links. Examples exist in the financial systems where the firms and banks (i.e. nodes) still exist but many financial trades (i.e. links) are gone during the crisis, and the air transportation systems where the airports (i.e. nodes) are still functional but many airlines (i.e. links) stop operating during bad weather. In this letter, we develop a link cascade model in complex networks. With this model, we find that both artificial and real networks tend to collapse even if a few links are initially attacked. However, the link cascading process can be effectively terminated by setting a few strong nodes in the network which do not respond to any link reduction. Finally, a simulated annealing algorithm is used to optimize the location of these strong nodes, which significantly improves the robustness of the networks against the link cascade. - Highlights: • We propose a link cascade model in complex networks. • Both artificial and real networks tend to collapse even if a few links are initially attacked. • The link cascading process can be effectively terminated by setting a few strong nodes. • A simulated annealing algorithm is used to optimize the location of these strong nodes.

  10. In vitro screening of reversible and time-dependent inhibition on CYP3A by TM208 and TM209 in rat liver microsomes

    Directory of Open Access Journals (Sweden)

    Miaoran Ning

    2012-04-01

    Full Text Available TM208 and TM209, dithiocarbamate derivatives with potential anti-cancer effects, were evaluated in reversible and time-dependent cytochrome P450 (CYP 3A inhibition assays in rat liver microsomes using testosterone as probe substrate. Both compounds were found to be weak reversible inhibitors and moderate mechanism-based inhibitors of rat CYP3A. For reversible inhibition on rat CYP3A, the Ki values of competitive inhibition model were 12.10±1.75 and 13.94±1.31 μM, respectively. For time-dependent inhibition, the inactivation constants (Kl were 31.93±12.64 and 32.91±15.58 μM, respectively, and the maximum inactivation rates (kinact were 0.03497±0.0069 and 0.07259±0.0172 min−1 respectively. These findings would provide useful in vitro information for future in vivo DDI studies on TM208 or TM209.

  11. In Vitro and In Vivo Evaluation of Polyherbal Formulation against Russell’s Viper and Cobra Venom and Screening of Bioactive Components by Docking Studies

    Directory of Open Access Journals (Sweden)

    G. Sakthivel

    2013-01-01

    Full Text Available The present study emphasizes to reveal the antivenom activity of Aristolochia bracteolata Lam., Tylophora indica (Burm.f. Merrill, and Leucas aspera S. which were evaluated against venoms of Daboia russelli russelli (Russell’s viper and Naja naja (Indian cobra. The aqueous extracts of leaves and roots of the above-mentioned plants and their polyherbal (1 : 1 : 1 formulation at a dose of 200 mg/kg showed protection against envenomed mice with LD50 doses of 0.44 mg/kg and 0.28 mg/kg against Russell’s viper and cobra venom, respectively. In in vitro antioxidant activities sample extracts showed free radical scavenging effects in dose dependent manner. Computational drug design and docking studies were carried out to predict the neutralizing principles of type I phospholipase A2 (PLA2 from Indian common krait venom. This confirmed that aristolochic acid and leucasin can neutralize type I PLA2 enzyme. Results suggest that these plants could serve as a source of natural antioxidants and common antidote for snake bite. However, further studies are needed to identify the lead molecule responsible for antidote activity.

  12. Characterisation and cytotoxic screening of metal oxide nanoparticles putative of interest to oral healthcare formulations in non-keratinised human oral mucosa cells in vitro.

    Science.gov (United States)

    Best, M; Phillips, G; Fowler, C; Rowland, J; Elsom, J

    2015-12-25

    Nanoparticles are increasingly being utilised in the innovation of consumer product formulations to improve their characteristics; however, established links between their properties, dose and cytotoxicity are not well defined. The purpose of this study was to screen four different nanomaterials of interest to oral care product development in the absence of stabilisers, alongside their respective bulk equivalents, within a non-keratinised oral epithelial cell model (H376). Particle morphology and size were characterised using scanning electron microscopy (SEM) and dynamic light scattering (DLS). The H376 model showed that zinc oxide (ZnO) was the most cytotoxic material at concentrations exceeding 0.031% w/v, as assessed using the lactate dehydrogenase (LDH) and dimethylthiazolyl-diphenyl-tetrazolium-bromide (MTT) assays. ZnO cytotoxicity does not appear to be dependent upon size of the particle; a result supported by SEM of cell-particle interactions. Differences in cytotoxicity were observed between the bulk and nanomaterial forms of hydroxyapatite and silica (SiO2); titanium dioxide (TiO2) was well tolerated in both forms at the doses tested. Overall, nano-size effects have some impact on the cytotoxicity of a material; however, these may not be as significant as chemical composition or surface properties. Our data highlights the complexities involved at the nano-scale, in both the characterisation of materials and in relation to cytotoxic properties exerted on oral epithelial cells. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Vessel segmentation in screening mammograms

    Science.gov (United States)

    Mordang, J. J.; Karssemeijer, N.

    2015-03-01

    Blood vessels are a major cause of false positives in computer aided detection systems for the detection of breast cancer. Therefore, the purpose of this study is to construct a framework for the segmentation of blood vessels in screening mammograms. The proposed framework is based on supervised learning using a cascade classifier. This cascade classifier consists of several stages where in each stage a GentleBoost classifier is trained on Haar-like features. A total of 30 cases were included in this study. In each image, vessel pixels were annotated by selecting pixels on the centerline of the vessel, control samples were taken by annotating a region without any visible vascular structures. This resulted in a total of 31,000 pixels marked as vascular and over 4 million control pixels. After training, the classifier assigns a vesselness likelihood to the pixels. The proposed framework was compared to three other vessel enhancing methods, i) a vesselness filter, ii) a gaussian derivative filter, and iii) a tubeness filter. The methods were compared in terms of area under the receiver operating characteristics curves, the Az values. The Az value of the cascade approach is 0:85. This is superior to the vesselness, Gaussian, and tubeness methods, with Az values of 0:77, 0:81, and 0:78, respectively. From these results, it can be concluded that our proposed framework is a promising method for the detection of vessels in screening mammograms.

  14. "Gold nanoparticles composite-folic acid conjugated graphene oxide nanohybrids" for targeted chemo-thermal cancer ablation: In vitro screening and in vivo studies.

    Science.gov (United States)

    Chauhan, Gaurav; Chopra, Vianni; Tyagi, Amit; Rath, Goutam; Sharma, Rakesh K; Goyal, Amit K

    2017-01-01

    Nano-graphene oxide (GO) nanometal composite (specifically nanogold and nanosilver) have shown to be a promising material for anticancer therapeutics. Owing to their high drug loading capacity, photothermal and synergizing effects, it is very important to exploit them for targeted chemo-thermal cancer therapeutics. In this work, gold nanoparticles (AuNPs) were selected as the composite metal, folic acid (FA) was taken as GO surface functionalization moiety for active tumor targeting of model anticancer drug Doxorubicin (Dox). AuNPs composite-folate conjugated graphene oxide (FA-GO@Au) nano-platforms were synthesized and characterized in detail. Near-infrared (NIR) sensitivity resulted in an aggravated release of both Dox and ionic gold from the nanohybrid surface. Simultaneous delivery of Dox and AuNPs in the cellular vicinity was further enhanced after localized NIR exposure which resulted in significantly improved cancer cell toxicity. Mechanistic evaluation revealed G0/G1 phase arrest due to increased DNA intercalation and provoked early apoptosis under NIR influence. Pharmacokinetics and organ distribution studies were carried out in healthy mice and rabbits to estimate the actual bio fate of these nanohybrids. In vivo studies showed substantial tumor regression in solid tumor model in Balb/c mice and NIR exposure induced photo-thermal effects further resulted in better tumor management. Study provides substantial evidences both at in vitro and in vivo level to support the fact that NIR induced local photo-thermal effects can solely be used as a tumor targeting tool. This NIR dependent nanohybrid approach presents a precise and flexible strategy for targeted chemotherapy and photo-thermal tumor ablation. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. β-Amyloid neurotoxicity is mediated by a glutamate-triggered excitotoxic cascade in rat nucleus basalis

    NARCIS (Netherlands)

    Harkany, T.; Ábrahám, I.; Timmerman, W.; Laskay, G.; Tóth, B.; Sasvári, M.; Kónya, C.; Sebens, J.B.; Korf, J.; Nyakas, C.; Zarándi, M.; Soós, K.; Penke, B.; Luiten, P.G.M.

    2000-01-01

    Whereas a cardinal role for β-amyloid protein (Aβ) has been postulated as a major trigger of neuronal injury in Alzheimer's disease, the pathogenic mechanism by which Aβ deranges nerve cells remains largely elusive. Here we report correlative in vitro and in vivo evidence that an excitotoxic cascade

  16. In vitro antimycobacterial activity and HPLC-DAD screening of phenolics from Ficus benjamina L. and Ficus luschnathiana (Miq.) Miq. leaves.

    Science.gov (United States)

    da Cruz, Ritiel Corrêa; Agertt, Vanessa; Boligon, Aline Augusti; Janovik, Vanessa; Anraku de Campos, Marli Matiko; Guillaume, Dominique; Athayde, Margareth Linde

    2012-01-01

    The total phenolic content (Folin-Ciocalteu) of the leaves of Ficus benjamina and Ficus luschnathiana was evaluated and screened by HPLC-DAD. Ficus luschnathiana crude extract (CE) presented phenolic content higher than that of F. benjamina (149.92 ± 3.65 versus 122.63 ± 2.79 mg of GAE). Kaempferol (1.63 ± 0.16 mg g(-1) dry weight of CE) and chlorogenic acid (17.77 ± 0.57 mg g(-1) of butanolic fraction) were identified and quantified in F. benjamina, whereas rutin (1.39 ± 0.20 mg g(-1)), caffeic (1.14 ± 0.13 mg g(-1)) and chlorogenic (3.73 ± 0.29 mg g(-1)) acids were quantified in the CE of F. luschnathiana. Additionaly, rutin (15.55 ± 1.92 mg g(-1)) and quercetin (3.53 ± 0.12 mg g(-1)) were quantified in ethyl acetate and butanolic fractions, respectively. Antimycobacterial activity of CEs and fractions was evaluated against Mycobacterium smegmatis by broth microdilution method. Ethyl acetate fraction from F. benjamina and n-butanol fraction from F. luschnathiana displayed the highest inhibitory activity (MIC = 312.50 µg mL(-1) and 156.25 µg mL(-1), respectively). Further studies are required to identify the compounds directly related to antimycobacterial activity.

  17. Diagnostic Accuracy of Loop-mediated Isothermal Amplifica-tion Assay as a Field Molecular Tool for Rapid Mass Screening of Old World Leishmania Infections in Sand Flies and In Vitro Culture

    Directory of Open Access Journals (Sweden)

    Mehdi GHODRATI

    2017-12-01

    Full Text Available AbstractBackground: We employed a highly sensitive loop-mediated isothermal amplification (LAMP by targeting 18S rRNA gene to identify the rapid mass screening of Leishmania infections in captured sand flies of southwest Iran and In vitro culture. Methods: One hundred fifty sand flies were collected from 11 sites adjacent to Iraqi’s borders in southern parts of Khuzestan Province by using sticky sheets of paper and CDC miniature light traps during late May 2014 to Nov 2015. Following morphological identification of sand flies species, the DNA of infected samples was extracted and amplified by PCR and LAMP assays by targeting ITS-rDNA and 18S rRNA genes. The PCR amplicons were directly sequenced to conduct the phylogenetic analysis Results: Ten (6.6% Leishmania infections were identified by LAMP assay (detection limit 0.01 parasites DNA among infected Sergentomyia baghdadis, S. sintoni and Phlebotomus papatasi sand flies that was more sensitive than PCR (n=6.4%; (detection limit 101parasites DNA. LAMP can identify 101-106promastigotes/100 µl RPMI 1640 while PCR recognized104-106 promastigotes. The majority infection rate of sand flies was confirmed to L. major inferred by phylogenetic analysis. Conclusion: This is the first exploration characterized the Old World Leishmania infections by LAMP technique in both infected sand flies and In vitro conditions. The LAMP method because of its shorter reaction time, robustness, more sensitivity, lack of requirement of complicated equipment and visual discriminatory of positivity can be appeared a promising tool instead of PCR to identify low Leishmania loads and entomological monitoring of leishmaniasis in resource-limited endemic of the world.

  18. Evaluation of the Isoniazid Preventive Therapy Care Cascade Among HIV-Positive Female Sex Workers in Mombasa, Kenya.

    Science.gov (United States)

    LaCourse, Sylvia M; Deya, Ruth W; Graham, Susan M; Masese, Linnet N; Jaoko, Walter; Mandaliya, Kischorchandra N; Overbaugh, Julie; McClelland, R Scott

    2017-09-01

    Kenyan female sex workers (FSWs) have a high HIV prevalence, increasing their tuberculosis (TB) risk. Despite recommendations that HIV-positive individuals be offered isoniazid preventive therapy (IPT), uptake has been limited. In this longitudinal cohort of HIV-positive FSWs, we retrospectively characterized the IPT care cascade between March 2000 and January 2010, including reasons for cascade loss or appropriate exit. Cascade success required completion of 6 months of IPT. Baseline characteristics were assessed as potential correlates of cascade loss using multivariable logistic regression. Among 642 HIV-positive FSWs eligible for IPT evaluation, median age was 31 years (IQR 26-35) with median CD4 lymphocyte count of 409 (IQR 292-604) cells per cubic millimeter. There were 249 (39%) women who successfully completed 6 months of IPT, 157 (24%) appropriately exited the cascade, and 236 (37%) were cascade losses. Most cascade losses occurred at symptom screen (38%, 90/236), chest radiograph evaluation (28%, 66/236), or during IPT treatment (30%, 71/236). Twenty-nine women were diagnosed with tuberculosis, including one after IPT initiation. Most women initiating IPT completed the course (71%, 249/351); 35 years; adjusted odds ratio (AOR) 2.65, 95% confidence interval (CI): 1.46 to 4.80 and AOR 1.78, 95% CI: 1.13 to 2.80, respectively], and those evaluated for IPT after antiretroviral availability in 2004 (AOR 1.92, 95% CI: 1.31 to 2.81), were more likely to be cascade losses. Implementation of IPT among HIV-positive FSWs in Kenya is feasible. However, significant losses along the IPT care cascade underscore the need for strategies improving retention in care.

  19. Discovery of potent, novel, non-toxic anti-malarial compounds via quantum modelling, virtual screening and in vitro experimental validation

    Directory of Open Access Journals (Sweden)

    Kaludov Nikola

    2011-09-01

    Full Text Available Abstract Background Developing resistance towards existing anti-malarial therapies emphasize the urgent need for new therapeutic options. Additionally, many malaria drugs in use today have high toxicity and low therapeutic indices. Gradient Biomodeling, LLC has developed a quantum-model search technology that uses quantum similarity and does not depend explicitly on chemical structure, as molecules are rigorously described in fundamental quantum attributes related to individual pharmacological properties. Therapeutic activity, as well as toxicity and other essential properties can be analysed and optimized simultaneously, independently of one another. Such methodology is suitable for a search of novel, non-toxic, active anti-malarial compounds. Methods A set of innovative algorithms is used for the fast calculation and interpretation of electron-density attributes of molecular structures at the quantum level for rapid discovery of prospective pharmaceuticals. Potency and efficacy, as well as additional physicochemical, metabolic, pharmacokinetic, safety, permeability and other properties were characterized by the procedure. Once quantum models are developed and experimentally validated, the methodology provides a straightforward implementation for lead discovery, compound optimizzation and de novo molecular design. Results Starting with a diverse training set of 26 well-known anti-malarial agents combined with 1730 moderately active and inactive molecules, novel compounds that have strong anti-malarial activity, low cytotoxicity and structural dissimilarity from the training set were discovered and experimentally validated. Twelve compounds were identified in silico and tested in vitro; eight of them showed anti-malarial activity (IC50 ≤ 10 μM, with six being very effective (IC50 ≤ 1 μM, and four exhibiting low nanomolar potency. The most active compounds were also tested for mammalian cytotoxicity and found to be non-toxic, with a

  20. Robustness of localized DNA strand displacement cascades.

    Science.gov (United States)

    Teichmann, Mario; Kopperger, Enzo; Simmel, Friedrich C

    2014-08-26

    Colocalization can strongly alter the kinetics and efficiency of chemical processes. For instance, in DNA-templated synthesis unfavorable reactions are sped up by placing reactants into close proximity onto a DNA scaffold. In biochemistry, clustering of enzymes has been demonstrated to enhance the reaction flux through some enzymatic cascades. Here we investigate the effect of colocalization on the performance of DNA strand displacement (DSD) reactions, an important class of reactions utilized in dynamic DNA nanotechnology. We study colocalization by immobilizing a two-stage DSD reaction cascade comprised of a “sender” and a “receiver” gate onto a DNA origami platform. The addition of a DNA (or RNA) input strand displaces a signal strand from the sender gate, which can then transfer to the receiver gate. The performance of the cascade is found to vary strongly with the distance between the gates. A cascade with an intermediate gate distance of ≈20 nm exhibits faster kinetics than those with larger distances, whereas a cascade with smaller distance is corrupted by excessive intraorigami leak reactions. The 20 nm cascade is found to be considerably more robust with respect to a competing reaction, and implementation of multiple receiver gates further increases this robustness. Our results indicate that for the 20 nm distance a fraction of signal strands is transferred locally to a receiver gate on the same platform, probably involving direct physical contact between the gates. The performance of the cascade is consistent with a simple model that takes “local” and “global” transfer processes into account.

  1. Preliminary Phytochemical Screening and In Vitro Anti-Helicobacter pylori Activity of Extracts of the Stem Bark of Bridelia micrantha (Hochst., Baill., Euphorbiaceae

    Directory of Open Access Journals (Sweden)

    Pascal O. Bessong

    2011-07-01

    Full Text Available Helicobacter pylori is a major risk factor for gastritis, ulcers and gastric cancer. This study was aimed to determine the antimicrobial activity of the stem bark of Bridelia. micrantha on H. pylori isolated in South Africa. Extracts and clarithromycin were tested against 31 clinical strains, including a standard strain (NCTC 11638 of H. pylori, by measuring the diameters of the corresponding inhibition zones, followed by determination of the Minimum Inhibitory Concentration (MIC (using metronidazole, and amoxicillin as control antibiotics and the rate of kill. Preliminary phytochemical screening was also done. Inhibition zone diameters which ranged from 0–23 mm were observed for all five of the extracts and 0–35 mm for clarithromycin. Marked susceptibility of strains (100% was noted for the acetone extract (P < 0.05, followed by ethyl acetate extract (93.5%. The MIC50 values ranged from 0.0048 to 0.156 mg/mL for the ethyl acetate extract and 0.0048 to 0.313 mg/mL for the acetone extract. The MIC90 values ranged from 0.0048 to 2.5 mg/mL for the ethyl acetate extract and 0.078 to >0.625 mg/mL for the acetone extract, respectively. Insignificant statistical difference in potency was observed when comparing the crude ethyl acetate extract to metronidazole and amoxicillin (P > 0.05. Complete killing of strain PE430C by the ethyl acetate extract was observed at 0.1 mg/mL (2 × MIC and 0.2 mg/mL (4 × MIC at 66 and 72 h. For strain PE369C, 100% killing was observed at 0.1 mg/mL (2 × MIC in 66 and 72 h. The ethyl acetate extract could thus be a potential source of lead molecules for the design of new anti-Helicobacter pylori therapies as this study further confirmed the presence of phytochemicals including alkaloids, flavonoids, steroids, tannins and saponins.

  2. Gene expression analysis of troglitazone reveals its impact on multiple pathways in cell culture: a case for in vitro platforms combined with gene expression analysis for early (idiosyncratic) toxicity screening.

    Science.gov (United States)

    Vansant, Gordon; Pezzoli, Patrick; Saiz, Robert; Birch, Aaron; Duffy, Chris; Ferre, Francois; Monforte, Joseph

    2006-01-01

    Peroxisome proliferator-activated receptor gamma (PPARgamma) agonists of the thiazolidinedione family are used for the treatment of type 2 diabetes mellitus due to their ability to reduce glucose and lipid levels in patients with this disease. Three thiazolidinediones that were approved for treatment are Rezulin (troglitazone), Avandia (rosiglitazone), and Actos (pioglitazone). Troglitazone was withdrawn from the market due to idiosyncratic drug toxicity. Rosiglitazone and pioglitazone are still on the market for the treatment of type 2 diabetes. The authors present data from a gene expression screen that compares the impact these three compounds have in rats, in rat hepatocytes, and in the clone 9 rat liver cell line. The authors monitored the changes in expression in multiple genes, including those related to xenobiotic metabolism, proliferation, DNA damage, oxidative stress, apoptosis, and inflammation. Compared to the other two compounds, troglitazone had a significant impact on many of the pathways monitored in vitro although no major perturbation was detected in vivo. The changes detected predict not only general toxicity but potential mechanisms of toxicity. Based on gene expression analysis, the authors propose there is not just one but multiple ways troglitazone could be toxic, depending on a patient's environment and genetic makeup, including immune response-related toxicity.

  3. Toxicology screen

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003578.htm Toxicology screen To use the sharing features on this page, please enable JavaScript. A toxicology screen refers to various tests that determine the ...

  4. Carrier Screening

    Science.gov (United States)

    ... pregnant are offered carrier screening for cystic fibrosis, hemoglobinopathies , and spinal muscular atrophy . You can have screening ... caused by a change in genes or chromosomes. Hemoglobinopathies: Any inherited disorder caused by changes in the ...

  5. Airport Screening

    Science.gov (United States)

    Health Physics Society Specialists in Radiation Safety Airport Screening Fact Sheet Adopted: May 2011 Photo courtesy of Dan Paluska/Flickr Denver Airport Security Screening Introduction With air travel regaining popularity and ...

  6. MRSA Screening

    Science.gov (United States)

    ... Links Patient Resources For Health Professionals Subscribe Search MRSA Screening Send Us Your Feedback Choose Topic At ... Related Content Related Images View Sources Formal Name Methicillin-resistant Staphylococcus aureus Screening This article was last reviewed on February ...

  7. MAP kinase cascades in Arabidopsis innate immunity

    DEFF Research Database (Denmark)

    Rasmussen, Magnus Wohlfahrt; Roux, Milena Edna; Petersen, Morten

    2012-01-01

    Plant mitogen-activated protein kinase (MAPK) cascades generally transduce extracellular stimuli into cellular responses. These stimuli include the perception of pathogen-associated molecular patterns (PAMPs) by host transmembrane pattern recognition receptors which trigger MAPK-dependent innate...... immune responses. In the model Arabidopsis, molecular genetic evidence implicates a number of MAPK cascade components in PAMP signaling, and in responses to immunity-related phytohormones such as ethylene, jasmonate, and salicylate. In a few cases, cascade components have been directly linked...... to the transcription of target genes or to the regulation of phytohormone synthesis. Thus MAPKs are obvious targets for bacterial effector proteins and are likely guardees of resistance proteins, which mediate defense signaling in response to the action of effectors, or effector-triggered immunity. This mini...

  8. Bifurcations analysis of turbulent energy cascade

    Energy Technology Data Exchange (ETDEWEB)

    Divitiis, Nicola de, E-mail: n.dedivitiis@gmail.com

    2015-03-15

    This note studies the mechanism of turbulent energy cascade through an opportune bifurcations analysis of the Navier–Stokes equations, and furnishes explanations on the more significant characteristics of the turbulence. A statistical bifurcations property of the Navier–Stokes equations in fully developed turbulence is proposed, and a spatial representation of the bifurcations is presented, which is based on a proper definition of the fixed points of the velocity field. The analysis first shows that the local deformation can be much more rapid than the fluid state variables, then explains the mechanism of energy cascade through the aforementioned property of the bifurcations, and gives reasonable argumentation of the fact that the bifurcations cascade can be expressed in terms of length scales. Furthermore, the study analyzes the characteristic length scales at the transition through global properties of the bifurcations, and estimates the order of magnitude of the critical Taylor-scale Reynolds number and the number of bifurcations at the onset of turbulence.

  9. Seeded QED cascades in counterpropagating laser pulses.

    Science.gov (United States)

    Grismayer, T; Vranic, M; Martins, J L; Fonseca, R A; Silva, L O

    2017-02-01

    The growth rates of seeded QED cascades in counterpropagating lasers are calculated with first-principles two- and three-dimensional QED-PIC (particle-in-cell) simulations. The dependence of the growth rate on the laser polarization and intensity is compared with analytical models that support the findings of the simulations. The models provide insight regarding the qualitative trend of the cascade growth when the intensity of the laser field is varied. A discussion about the cascade's threshold is included, based on the analytical and numerical results. These results show that relativistic pair plasmas and efficient conversion from laser photons to γ rays can be observed with the typical intensities planned to operate on future ultraintense laser facilities such as ELI or Vulcan.

  10. Emergence of event cascades in inhomogeneous networks

    CERN Document Server

    Onaga, Tomokatsu

    2016-01-01

    There is a commonality among contagious diseases, tweets, urban crimes, nuclear reactions, and neuronal firings that past events facilitate the future occurrence of events. The spread of events has been extensively studied such that the systems exhibit catastrophic chain reactions if the interaction represented by the ratio of reproduction exceeds unity; however, their subthreshold states for the case of the weaker interaction are not fully understood. Here, we report that these systems are possessed by nonstationary cascades of event-occurrences already in the subthreshold regime. Event cascades can be harmful in some contexts, when the peak-demand causes vaccine shortages, heavy traffic on communication lines, frequent crimes, or large fluctuations in nuclear reactions, but may be beneficial in other contexts, such that spontaneous activity in neural networks may be used to generate motion or store memory. Thus it is important to comprehend the mechanism by which such cascades appear, and consider controlli...

  11. Epidemic and Cascading Survivability of Complex Networks

    DEFF Research Database (Denmark)

    Manzano, Marc; Calle, Eusebi; Ripoll, Jordi

    2014-01-01

    Our society nowadays is governed by complex networks, examples being the power grids, telecommunication networks, biological networks, and social networks. It has become of paramount importance to understand and characterize the dynamic events (e.g. failures) that might happen in these complex...... networks. For this reason, in this paper, we propose two measures to evaluate the vulnerability of complex networks in two different dynamic multiple failure scenarios: epidemic-like and cascading failures. Firstly, we present epidemic survivability ( ES ), a new network measure that describes...... the vulnerability of each node of a network under a specific epidemic intensity. Secondly, we propose cascading survivability ( CS ), which characterizes how potentially injurious a node is according to a cascading failure scenario. Then, we show that by using the distribution of values obtained from ES and CS...

  12. Epidemic and Cascading Survivability of Complex Networks

    CERN Document Server

    Manzano, Marc; Ripoll, Jordi; Fagertun, Anna Manolova; Torres-Padrosa, Victor; Pahwa, Sakshi; Scoglio, Caterina

    2014-01-01

    Our society nowadays is governed by complex networks, examples being the power grids, telecommunication networks, biological networks, and social networks. It has become of paramount importance to understand and characterize the dynamic events (e.g. failures) that might happen in these complex networks. For this reason, in this paper, we propose two measures to evaluate the vulnerability of complex networks in two different dynamic multiple failure scenarios: epidemic-like and cascading failures. Firstly, we present \\emph{epidemic survivability} ($ES$), a new network measure that describes the vulnerability of each node of a network under a specific epidemic intensity. Secondly, we propose \\emph{cascading survivability} ($CS$), which characterizes how potentially injurious a node is according to a cascading failure scenario. Then, we show that by using the distribution of values obtained from $ES$ and $CS$ it is possible to describe the vulnerability of a given network. We consider a set of 17 different compl...

  13. Cancer Screening

    OpenAIRE

    Krishna Prasad

    2004-01-01

    Cancer screening is a means to detect cancer early with the goal of decreasing morbidity and mortality. At present, there is a reasonable consensus regarding screening for breast, cervical and colorectal cances and the role of screening is under trial in case of cancers of the lung,  ovaries and prostate. On the other hand, good screening tests are not available for some of the commonest cancers in India like the oral, pharyngeal, esophageal and stomach cancers.

  14. Piscivores, Trophic Cascades, and Lake Management

    Directory of Open Access Journals (Sweden)

    Ray W. Drenner

    2002-01-01

    Full Text Available The concept of cascading trophic interactions predicts that an increase in piscivore biomass in lakes will result in decreased planktivorous fish biomass, increased herbivorous zooplankton biomass, and decreased phytoplankton biomass. Though often accepted as a paradigm in the ecological literature and adopted by lake managers as a basis for lake management strategies, the trophic cascading interactions hypothesis has not received the unequivocal support (in the form of rigorous experimental testing that might be expected of a paradigm. Here we review field experiments and surveys, testing the hypothesis that effects of increasing piscivore biomass will cascade down through the food web yielding a decline in phytoplankton biomass. We found 39 studies in the scientific literature examining piscivore effects on phytoplankton biomass. Of the studies, 22 were confounded by supplemental manipulations (e.g., simultaneous reduction of nutrients or removal of planktivores and could not be used to assess piscivore effects. Of the 17 nonconfounded studies, most did not find piscivore effects on phytoplankton biomass and therefore did not support the trophic cascading interactions hypothesis. However, the trophic cascading interactions hypothesis also predicts that lake systems containing piscivores will have lower phytoplankton biomass for any given phosphorus concentration. Based on regression analyses of chlorophyll�total phosphorus relationships in the 17 nonconfounded piscivore studies, this aspect of the trophic cascading interactions hypothesis was supported. The slope of the chlorophyll vs. total phosphorus regression was lower in lakes with planktivores and piscivores compared with lakes containing only planktivores but no piscivores. We hypothesize that this slope can be used as an indicator of “functional piscivory” and that communities with extremes of functional piscivory (zero and very high represent classical 3- and 4-trophic level

  15. Volcano geodesy in the Cascade arc, USA

    Science.gov (United States)

    Poland, Michael; Lisowski, Michael; Dzurisin, Daniel; Kramer, Rebecca; McLay, Megan; Pauk, Benjamin

    2017-01-01

    Experience during historical time throughout the Cascade arc and the lack of deep-seated deformation prior to the two most recent eruptions of Mount St. Helens might lead one to infer that Cascade volcanoes are generally quiescent and, specifically, show no signs of geodetic change until they are about to erupt. Several decades of geodetic data, however, tell a different story. Ground- and space-based deformation studies have identified surface displacements at five of the 13 major Cascade arc volcanoes that lie in the USA (Mount Baker, Mount St. Helens, South Sister, Medicine Lake, and Lassen volcanic center). No deformation has been detected at five volcanoes (Mount Rainier, Mount Hood, Newberry Volcano, Crater Lake, and Mount Shasta), and there are not sufficient data at the remaining three (Glacier Peak, Mount Adams, and Mount Jefferson) for a rigorous assessment. In addition, gravity change has been measured at two of the three locations where surveys have been repeated (Mount St. Helens and Mount Baker show changes, while South Sister does not). Broad deformation patterns associated with heavily forested and ice-clad Cascade volcanoes are generally characterized by low displacement rates, in the range of millimeters to a few centimeters per year, and are overprinted by larger tectonic motions of several centimeters per year. Continuous GPS is therefore the best means of tracking temporal changes in deformation of Cascade volcanoes and also for characterizing tectonic signals so that they may be distinguished from volcanic sources. Better spatial resolution of volcano deformation can be obtained through the use of campaign GPS, semipermanent GPS, and interferometric synthetic aperture radar observations, which leverage the accumulation of displacements over time to improve signal to noise. Deformation source mechanisms in the Cascades are diverse and include magma accumulation and withdrawal, post-emplacement cooling of recent volcanic deposits, magmatic

  16. Cascade morphology transition in bcc metals.

    Science.gov (United States)

    Setyawan, Wahyu; Selby, Aaron P; Juslin, Niklas; Stoller, Roger E; Wirth, Brian D; Kurtz, Richard J

    2015-06-10

    Energetic atom collisions in solids induce shockwaves with complex morphologies. In this paper, we establish the existence of a morphological transition in such cascades. The order parameter of the morphology is defined as the exponent, b, in the defect production curve as a function of cascade energy (N(F) ~ E(MD)(b)). Response of different bcc metals can be compared in a consistent energy domain when the energy is normalized by the transition energy, μ, between the high- and the low-energy regime. Using Cr, Fe, Mo and W data, an empirical formula of μ as a function of displacement threshold energy, E(d), is presented for bcc metals.

  17. Cascade morphology transition in bcc metals

    OpenAIRE

    Setyawan, Wahyu; Selby, Aaron P.; Juslin, Niklas; Stoller, Roger E; Wirth, Brian D.; Kurtz, Richard J.

    2014-01-01

    Energetic atom collisions in solids induce shockwaves with complex morphologies. In this paper, we establish the existence of a morphological transition in such cascades. The order parameter of the morphology is defined as the exponent, $b$, in the defect production curve as a function of cascade energy ($N_F \\sim E_{MD}^b$). Response of different bcc metals can be compared in a consistent energy domain when the energy is normalized by the transition energy, $\\mu$, between the high- and the l...

  18. Robustness of networks against cascading failures

    Science.gov (United States)

    Dou, Bing-Lin; Wang, Xue-Guang; Zhang, Shi-Yong

    2010-06-01

    Inspired by other related works, this paper proposes a non-linear load-capacity model against cascading failures, which is more suitable for real networks. The simulation was executed on the B-A scale-free network, E-R random network, Internet AS level network, and the power grid of the western United States. The results show that the model is feasible and effective. By studying the relationship between network cost and robustness, we find that the model can defend against cascading failures better and requires a lower investment cost when higher robustness is required.

  19. Robustness of network controllability in cascading failure

    Science.gov (United States)

    Chen, Shi-Ming; Xu, Yun-Fei; Nie, Sen

    2017-04-01

    It is demonstrated that controlling complex networks in practice needs more inputs than that predicted by the structural controllability framework. Besides, considering the networks usually faces to the external or internal failure, we define parameters to evaluate the control cost and the variation of controllability after cascades, exploring the effect of number of control inputs on the controllability for random networks and scale-free networks in the process of cascading failure. For different topological networks, the results show that the robustness of controllability will be stronger through allocating different control inputs and edge capacity.

  20. Volcano geodesy in the Cascade arc, USA

    Science.gov (United States)

    Poland, Michael P.; Lisowski, Michael; Dzurisin, Daniel; Kramer, Rebecca; McLay, Megan; Pauk, Ben

    2017-08-01

    Experience during historical time throughout the Cascade arc and the lack of deep-seated deformation prior to the two most recent eruptions of Mount St. Helens might lead one to infer that Cascade volcanoes are generally quiescent and, specifically, show no signs of geodetic change until they are about to erupt. Several decades of geodetic data, however, tell a different story. Ground- and space-based deformation studies have identified surface displacements at five of the 13 major Cascade arc volcanoes that lie in the USA (Mount Baker, Mount St. Helens, South Sister, Medicine Lake, and Lassen volcanic center). No deformation has been detected at five volcanoes (Mount Rainier, Mount Hood, Newberry Volcano, Crater Lake, and Mount Shasta), and there are not sufficient data at the remaining three (Glacier Peak, Mount Adams, and Mount Jefferson) for a rigorous assessment. In addition, gravity change has been measured at two of the three locations where surveys have been repeated (Mount St. Helens and Mount Baker show changes, while South Sister does not). Broad deformation patterns associated with heavily forested and ice-clad Cascade volcanoes are generally characterized by low displacement rates, in the range of millimeters to a few centimeters per year, and are overprinted by larger tectonic motions of several centimeters per year. Continuous GPS is therefore the best means of tracking temporal changes in deformation of Cascade volcanoes and also for characterizing tectonic signals so that they may be distinguished from volcanic sources. Better spatial resolution of volcano deformation can be obtained through the use of campaign GPS, semipermanent GPS, and interferometric synthetic aperture radar observations, which leverage the accumulation of displacements over time to improve signal to noise. Deformation source mechanisms in the Cascades are diverse and include magma accumulation and withdrawal, post-emplacement cooling of recent volcanic deposits, magmatic

  1. In vivo RNAi screen reveals neddylation genes as novel regulators of Hedgehog signaling.

    Directory of Open Access Journals (Sweden)

    Juan Du

    Full Text Available Hedgehog (Hh signaling is highly conserved in all metazoan animals and plays critical roles in many developmental processes. Dysregulation of the Hh signaling cascade has been implicated in many diseases, including cancer. Although key components of the Hh pathway have been identified, significant gaps remain in our understanding of the regulation of individual Hh signaling molecules. Here, we report the identification of novel regulators of the Hh pathway, obtained from an in vivo RNA interference (RNAi screen in Drosophila. By selectively targeting critical genes functioning in post-translational modification systems utilizing ubiquitin (Ub and Ub-like proteins, we identify two novel genes (dUba3 and dUbc12 that negatively regulate Hh signaling activity. We provide in vivo and in vitro evidence illustrating that dUba3 and dUbc12 are essential components of the neddylation pathway; they function in an enzyme cascade to conjugate the ubiquitin-like NEDD8 modifier to Cullin proteins. Neddylation activates the Cullin-containing ubiquitin ligase complex, which in turn promotes the degradation of Cubitus interruptus (Ci, the downstream transcription factor of the Hh pathway. Our study reveals a conserved molecular mechanism of the neddylation pathway in Drosophila and sheds light on the complex post-translational regulations in Hh signaling.

  2. Double screening

    Energy Technology Data Exchange (ETDEWEB)

    Gratia, Pierre [Department of Physics, University of Chicago,South Ellis Avenue, Chicago, IL 60637 (United States); Hu, Wayne [Department of Astronomy and Astrophysics, University of Chicago,South Ellis Avenue, Chicago, IL 60637 (United States); Enrico Fermi Institute and Kavli Institute for Cosmological Physics, University of Chicago,South Ellis Avenue, Chicago, IL 60637 (United States); Joyce, Austin [Enrico Fermi Institute and Kavli Institute for Cosmological Physics, University of Chicago,South Ellis Avenue, Chicago, IL 60637 (United States); Ribeiro, Raquel H. [School of Physics and Astronomy, Queen Mary University of London,Mile End Road, London, E1 4NS (United Kingdom)

    2016-06-15

    Attempts to modify gravity in the infrared typically require a screening mechanism to ensure consistency with local tests of gravity. These screening mechanisms fit into three broad classes; we investigate theories which are capable of exhibiting more than one type of screening. Specifically, we focus on a simple model which exhibits both Vainshtein and kinetic screening. We point out that due to the two characteristic length scales in the problem, the type of screening that dominates depends on the mass of the sourcing object, allowing for different phenomenology at different scales. We consider embedding this double screening phenomenology in a broader cosmological scenario and show that the simplest examples that exhibit double screening are radiatively stable.

  3. Neuronal necrosis is regulated by a conserved chromatin-modifying cascade

    Science.gov (United States)

    Liu, Kai; Ding, Lianggong; Li, Yuhong; Yang, Hui; Zhao, Chunyue; Lei, Ye; Han, Shuting; Tao, Wei; Miao, Dengshun; Steller, Hermann; Welsh, Michael J.; Liu, Lei

    2014-01-01

    Neuronal necrosis induced by calcium overload causes devastating brain dysfunction in diseases such as stroke and brain trauma. It has been considered a stochastic event lacking genetic regulation, and pharmacological means to suppress neuronal necrosis are lacking. Using a Drosophila model of calcium overloading, we found JIL-1/mitogen- and stress-activated protein kinase 1/2 is a regulator of neuronal necrosis through phosphorylation of histone H3 serine 28 (H3S28ph). Further, we identified its downstream events including displacement of polycomb repressive complex 1 (PRC1) and activation of Trithorax (Trx). To test the role of JIL-1/PRC1/Trx cascade in mammals, we studied the necrosis induced by glutamate in rat cortical neuron cultures and rodent models of brain ischemia and found the cascade is activated in these conditions and inhibition of the cascade suppresses necrosis in vitro and in vivo. Together, our research demonstrates that neuronal necrosis is regulated by a chromatin-modifying cascade, and this discovery may provide potential therapeutic targets and biomarkers for neuronal necrosis. PMID:25201987

  4. Gene regulation by MAP kinase cascades

    DEFF Research Database (Denmark)

    Fiil, Berthe Katrine; Petersen, Klaus; Petersen, Morten

    2009-01-01

    Mitogen-activated protein kinase (MAPK) cascades are signaling modules that transduce extracellular stimuli to a range of cellular responses. Research in yeast and metazoans has shown that MAPK-mediated phosphorylation directly or indirectly regulates the activity of transcription factors. Plant ...... gene expression....

  5. Multilevel Inverter by Cascading Industrial VSI

    DEFF Research Database (Denmark)

    Teodorescu, Remus; Blaabjerg, Frede; Pedersen, John Kim

    2002-01-01

    by a standard triphase IGBT inverter module. Thus, a high fault tolerance is being achieved and the output transformer requirement is eliminated. A staggered space-vector modulation technique applicable to three-phase cascaded voltage-source inverter topologies is also demonstrated. Both computer simulations...

  6. Criticality in a cascading failure blackout model

    Energy Technology Data Exchange (ETDEWEB)

    Nedic, Dusko P.; Kirschen, Daniel S. [University of Manchester (United Kingdom). School of Electrical and Electronic Engineering; Dobson, Ian [University of Wisconsin, Madison, WI (United States). ECE Department; Carreras, Benjamin A.; Lynch, Vickie E. [Oak Ridge National Laboratory, Oak Ridge, TN (United States)

    2006-11-15

    We verify and examine criticality in a 1000 bus network with an AC blackout model that represents many of the interactions that occur in cascading failure. At the critical loading there is a sharp rise in the mean blackout size and a power law probability distribution of blackout size that indicates a significant risk of large blackouts. (author)

  7. Geothermal research, Oregon Cascades: Final technical report

    Energy Technology Data Exchange (ETDEWEB)

    Priest, G.R.; Black, G.L.

    1988-10-27

    Previous USDOE-funded geothermal studies have produced an extensive temperature gradient and heat flow data base for the State of Oregon. One of the important features identified as a result of these studies is a rapid transition from heat flow values on the order of 40 mW/m/sup 2/ in the Willamette Valley and Western Cascades to values of greater than or equal to100 mW/m/sup 2/ in the High Cascades and the eastern portion of the Western Cascades. These data indicate that the Cascade Range in Oregon has potential as a major geothermal province and stimulated much of the later work completed by government agencies and private industry. Additional data generated as a result of this grant and published in DOGAMI Open-File Report 0-86-2 further define the location and magnitude of this transition zone. In addition, abundant data collected from the vicinity of Breitenbush and Austin Hot Springs have permitted the formulation of relatively detailed models of these hydrothermal systems. These models are published in DOGAMI Open-File Report 0-88-5. Task 1.2 of the Deliverables section of Amendment M001 is fulfilled by DOGAMI publication GMS-48, Geologic map of the McKenzie Bridge quadrangle, Lane County, Oregon. This map was printed in October, 1988, and is part of the final submission to USDOE. 8 refs.

  8. The Attention Cascade Model and Attentional Blink

    Science.gov (United States)

    Shih, Shui-I

    2008-01-01

    An attention cascade model is proposed to account for attentional blinks in rapid serial visual presentation (RSVP) of stimuli. Data were collected using single characters in a single RSVP stream at 10 Hz [Shih, S., & Reeves, A. (2007). "Attentional capture in rapid serial visual presentation." "Spatial Vision", 20(4), 301-315], and single words,…

  9. Quantum Cascade Lasers in Biomedical Infrared Imaging.

    Science.gov (United States)

    Bird, Benjamin; Baker, Matthew J

    2015-10-01

    Technological advances, namely the integration of quantum cascade lasers (QCLs) within an infrared (IR) microscope, are enabling the development of valuable label-free biomedical-imaging tools capable of targeting and detecting salient chemical species within practical clinical timeframes. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Nonconforming finite elements and the Cascade iteration

    NARCIS (Netherlands)

    Stevenson, R.

    1999-01-01

    We derive sucient conditions under which the Cascade iteration applied to nonconforming nite element discretizations yields an optimal solver. Key ingredients are optimal error estimates of such discretizations, which we therefore study in detail. We derive a new, ecient modied Morley nite element

  11. Cascade Training and Teachers' Professional Development.

    Science.gov (United States)

    Hayes, David

    2000-01-01

    Examines the experience of a nationwide inservice teacher development project in Sri Lanka that aims to remedy the potential deficiencies of cascade models of teacher development. Shows how project training and development strategies that are context sensitive, collaborative, and reflexive seek to involve teachers in managing their own…

  12. Cascading effects of overfishing marine systems

    NARCIS (Netherlands)

    Scheffer, M.; Carpenter, S.; Young, de B.

    2005-01-01

    Profound indirect ecosystem effects of overfishing have been shown for coastal systems such as coral reefs and kelp forests. A new study from the ecosystem off the Canadian east coast now reveals that the elimination of large predatory fish can also cause marked cascading effects on the pelagic food

  13. Modeling and simulation of cascading contingencies

    Science.gov (United States)

    Zhang, Jianfeng

    This dissertation proposes a new approach to model and study cascading contingencies in large power systems. The most important contribution of the work involves the development and validation of a heuristic analytic model to assess the likelihood of cascading contingencies, and the development and validation of a uniform search strategy. We model the probability of cascading contingencies as a function of power flow and power flow changes. Utilizing logistic regression, the proposed model is calibrated using real industry data. This dissertation analyzes random search strategies for Monte Carlo simulations and proposes a new uniform search strategy based on the Metropolis-Hastings Algorithm. The proposed search strategy is capable of selecting the most significant cascading contingencies, and it is capable of constructing an unbiased estimator to provide a measure of system security. This dissertation makes it possible to reasonably quantify system security and justify security operations when economic concerns conflict with reliability concerns in the new competitive power market environment. It can also provide guidance to system operators about actions that may be taken to reduce the risk of major system blackouts. Various applications can be developed to take advantage of the quantitative security measures provided in this dissertation.

  14. Cascade theory in isotopic separation processes; Theorie des cascades en separation isotopique

    Energy Technology Data Exchange (ETDEWEB)

    Agostini, J.P.

    1994-06-01

    Three main areas are developed within the scope of this work: - the first one is devoted to fundamentals: separative power, value function, ideal cascade and square cascade. Applications to two main cases are carried out, namely: Study of binary isotopic mix, Study of processes with a small enrichment coefficient. - The second one is devoted to cascade coupling -high-flux coupling (more widely used and better known) as well as low-flux coupling are presented and compared to one another. - The third one is an outlook on problems linked to cascade transients. Those problem are somewhat intricate and their interest lies mainly into two areas: economics where the start-up time may have a large influence on the interests paid during the construction and start-up period, military productions where the start-up time has a direct bearing on the production schedule. (author). 50 figs. 3 annexes. 12 refs. 6 tabs.

  15. Higher-order Kerr effect and harmonic cascading in gases

    OpenAIRE

    Bache, Morten; Eilenberger, Falk; Minardi, Stefano

    2012-01-01

    The higher-order Kerr effect (HOKE) has recently been advocated to explain measurements of the saturation of the nonlinear refractive index in gases. Here we show that cascaded third-harmonic generation results in an effective fifth-order nonlinearity that is negative and significant. Higher-order harmonic cascading will also occur from the HOKE, and the cascading contributions may significantly modify the observed nonlinear index change. At lower wavelengths, cascading increases the HOKE sat...

  16. A Modeling Framework for System Restoration from Cascading Failures

    OpenAIRE

    Chaoran Liu; Daqing Li; Enrico Zio; Rui Kang

    2014-01-01

    System restoration from cascading failures is an integral part of the overall defense against catastrophic breakdown in networked critical infrastructures. From the outbreak of cascading failures to the system complete breakdown, actions can be taken to prevent failure propagation through the entire network. While most analysis efforts have been carried out before or after cascading failures, restoration during cascading failures has been rarely studied. In this paper, we present a modeling f...

  17. Exploring cascading outages and weather via processing historic data

    OpenAIRE

    Dobson, Ian; Carrington, NichelleLe K.; Zhou, Kai; Wang, Zhaoyu; Carreras, Benjamin A.; Reynolds-Barredo, Jose M.

    2017-01-01

    We describe some bulk statistics of historical initial line outages and the implications for forming contingency lists and understanding which initial outages are likely to lead to further cascading. We use historical outage data to estimate the effect of weather on cascading via cause codes and via NOAA storm data. Bad weather significantly increases outage rates and interacts with cascading effects, and should be accounted for in cascading models and simulations. We suggest how weather effe...

  18. Improvement of banana cv. Rasthali (Silk, AAB) against Fusarium oxysporum f.sp. cubense (VCG 0124/5) through induced mutagenesis: Determination of LD50 specific to mutagen, explants, toxins and in vitro and in vivo screening for Fusarium wilt resistance.

    Science.gov (United States)

    Saraswathi, M S; Kannan, G; Uma, S; Thangavelu, R; Backiyarani, S

    2016-05-01

    Shoot tips and in vitro grown proliferating buds of banana cv. Rasthali (Silk, AAB) were treated with various concentrations and durations of chemical mutagens viz., EMS, NaN3 and DES. LD50 for shoot tips based on 50% reduction in fresh weight was determined as 2% for 3 h, 0.02% for 5 h and 0.15% for 5 h, while for proliferating buds, they were 0.6% for 30 min, 0.01% for 2 h and 0.06% for 2 h for the mutagens EMS, NaN3 and DES, respectively. Subsequently, the mutated explants were screened in vitro against fusarium wilt using selection agents like fusaric acid and culture filtrate. LD50 for in vitro selection agents calculated based on 50% survival of explants was 0.050 mM and 7% for fusaric acid and culture filtrate, respectively and beyond which a rapid decline in growth was observed. This was followed by pot screening which led to the identification of three putative resistant mutants with an internal disease score of 1 (corm completely clean, no vascular discolouration). The putative mutants identified in the present study have also been mass multiplied in vitro.

  19. Calculation of transonic flow in radial turbine blade cascade

    Science.gov (United States)

    Petr, Straka

    2017-09-01

    Numerical modeling of transonic centripetal turbulent flow in radial blade cascade is described in this paper. Attention is paid to effect of the outlet confusor on flow through the radial blade cascade. Parameters of presented radial blade cascade are compared with its linear representation

  20. Hollow Anode Cascading Plasma Focus | Alabraba | Journal of the ...

    African Journals Online (AJOL)

    Journal of the Nigerian Association of Mathematical Physics ... Using the 3-phase model for each focus event, the 9-phase, two solid disc auxiliary anode cascading plasma focus has been extended to include holes at the center of each cascade anode (hereafter referred to as hollow anode cascading focus) with a view of ...

  1. Genetic counseling and cascade genetic testing in Lynch syndrome.

    Science.gov (United States)

    Hampel, Heather

    2016-07-01

    Lynch syndrome is the most common cause of inherited colorectal and endometrial cancers. Individuals with Lynch syndrome have a 10-80 % lifetime risk for colorectal cancer and a 15-60 % lifetime risk for endometrial cancer. Both cancers are preventable through chemoprevention, intensive cancer surveillance, and risk-reducing surgery options. Efforts to identify as many individuals with Lynch syndrome as possible will prevent cancers and save lives. This includes the traditional cancer genetic counseling model whereby individuals with and without cancer are evaluated for a possible Lynch syndrome diagnosis based on their personal and family history of colon polyps and cancers. It also includes universal tumor screening for Lynch syndrome whereby all individuals with colorectal or endometrial cancer are screened for tumor features of Lynch syndrome at the time of diagnosis. Those with tumors suspicious for Lynch syndrome are referred for cancer genetic counseling regardless of their family history of cancer. This two approaches must be maximized to attain high patient reach. Finally, and perhaps most importantly, cascade testing among the at-risk relatives of those diagnosed with Lynch syndrome is critically important to maximize the diagnosis of individuals with Lynch syndrome. In fact, the cost-effectiveness of universal tumor screening for Lynch syndrome relies entirely on counseling and testing as many at-risk individuals as possible since young unaffected individuals stand to benefit the most from an early diagnosis of Lynch syndrome. This approach must be optimized to achieve high family reach. It will take a concerted effort from patients, clinicians and public health officials to improve current approaches to the diagnosis of Lynch syndrome and the prevention and treatment of Lynch syndrome-associated cancer but these lessons can be applied to other conditions as the ultimate example of personalized medicine.

  2. Quantum Cascade Lasers Modulation and Applications

    Science.gov (United States)

    Luzhansky, Edward

    The mid-wave IR (MWIR) spectral band, extending from 3 to 5 microns, is considered to be a low loss atmospheric window. There are several spectral sub-bands with relatively low atmospheric attenuation in this region making it popular for various commercial and military applications. Relatively low thermal and solar background emissions, effective penetration through the natural and anthropogenic obscurants and eye safety add to the long list of advantages of MWIR wavelengths. Quantum Cascade Lasers are compact semiconductor devices capable of operating in MWIR spectrum. They are based on inter-subband transitions in a multiple-quantum-well (QW) hetero-structure, designed by means of band-structure engineering. The inter-subband nature of the optical transition has several key advantages. First, the emission wavelength is primarily a function of the QW thickness. This characteristic allows choosing well-understood and reliable semiconductors for the generation of light in a wavelength range of interest. Second, a cascade process in which tens of photons are generated per injected electron. This cascading process is behind the intrinsic high-power capabilities of QCLs. This dissertation is focused on modulation properties of Quantum Cascade Lasers. Both amplitude and phase/frequency modulations were studied including modulation bandwidth, modulation efficiency and chirp linearity. Research was consisted of the two major parts. In the first part we describe the theory of frequency modulation (FM) response of Distributed Feedback Quantum Cascade Lasers (DFB QCL). It includes cascading effect on the QCL's maximum modulation frequency. The "gain levering" effect for the maximum FM response of the two section QCLs was studied as well. In the second part of research we concentrated on the Pulse Position Amplitude Modulation of a single section QCL. The low complexity, low size, weight and power Mid-Wavelength Infra-Red optical communications transceiver concept is

  3. Higher-order Kerr effect and harmonic cascading in gases

    DEFF Research Database (Denmark)

    Bache, Morten; Eilenberger, Falk; Minardi, Stefano

    2012-01-01

    The higher-order Kerr effect (HOKE) has recently been advocated to explain measurements of the saturation of the nonlinear refractive index in gases. Here we show that cascaded third-harmonic generation results in an effective fifth-order nonlinearity that is negative and significant. Higher......-order harmonic cascading will also occur from the HOKE, and the cascading contributions may significantly modify the observed nonlinear index change. At lower wavelengths, cascading increases the HOKE saturation intensity, while for longer wavelengths cascading will decrease the HOKE saturation intensity....

  4. Higher-order Kerr effect and harmonic cascading in gases.

    Science.gov (United States)

    Bache, Morten; Eilenberger, Falk; Minardi, Stefano

    2012-11-15

    The higher-order Kerr effect (HOKE) has recently been advocated to explain measurements of the saturation of the nonlinear refractive index in gases. Here we show that cascaded third-harmonic generation results in an effective fifth-order nonlinearity that is negative and significant. Higher-order harmonic cascading will also occur from the HOKE, and the cascading contributions may significantly modify the observed nonlinear index change. At lower wavelengths, cascading increases the HOKE saturation intensity, while for longer wavelengths cascading will decrease the HOKE saturation intensity.

  5. Developmental Screening

    Science.gov (United States)

    ... bye,” and pointing to something interesting are all developmental milestones, or things most children can do by a ... screening are ways to look for your child’s developmental milestones. Developmental Monitoring Developmental Screening WHO: You — parents, grandparents, ...

  6. Cascade morphology transition in bcc metals

    Energy Technology Data Exchange (ETDEWEB)

    Setyawan, Wahyu; Selby, A.; Juslin, Niklas; Stoller, Roger E.; Wirth, Brian D.; Kurtz, Richard J.

    2015-06-10

    Energetic atom collisions in solids induce shockwaves with complex morphologies. In this paper, we establish the existence of a morphological transition in such cascades. The order parameter of the morphology is defined as the exponent, $b$, in the defect production curve as a function of cascade energy ($N_F$$ \\sim$$E_{MD}^b$). Response of different bcc metals can be compared in a consistent energy domain when the energy is normalized by the transition energy, $\\mu$, between the high- and the low-energy regime. Using Cr, Fe, Mo and W data, an empirical formula of $\\mu$ as a function of displacement threshold energy, $E_d$, is presented for bcc metals.

  7. Cascade Apartments: Deep Energy Multifamily Retrofit

    Energy Technology Data Exchange (ETDEWEB)

    Gordon, A. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Mattheis, L. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Kunkle, R. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Howard, L. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Lubliner, M. [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2014-02-01

    In December of 2009-10, King County Housing Authority (KCHA) implemented energy retrofit improvements in the Cascade multifamily community, located in Kent, Washington (marine climate.)This research effort involved significant coordination from stakeholders KCHA, WA State Department of Commerce, utility Puget Sound Energy, and Cascade tenants. This report focuses on the following three primary BA research questions : 1. What are the modeled energy savings using DOE low income weatherization approved TREAT software? 2. How did the modeled energy savings compare with measured energy savings from aggregate utility billing analysis? 3. What is the Savings to Investment Ratio (SIR) of the retrofit package after considering utility window incentives and KCHA capitol improvement funding.

  8. Cascade Apartments: Deep Energy Multifamily Retrofit

    Energy Technology Data Exchange (ETDEWEB)

    Gordon, A. [Washington State Univ. Energy Program, Olympia, WA (United States); Mattheis, L. [Washington State Univ. Energy Program, Olympia, WA (United States); Kunkle, R. [Washington State Univ. Energy Program, Olympia, WA (United States); Howard, L. [Washington State Univ. Energy Program, Olympia, WA (United States); Lubliner, M. [Washington State Univ. Energy Program, Olympia, WA (United States)

    2014-02-01

    In December of 2009-10, King County Housing Authority (KCHA) implemented energy retrofit improvements in the Cascade multifamily community, located in Kent, Washington (marine climate.)This research effort involved significant coordination from stakeholders KCHA, WA State Department of Commerce, utility Puget Sound Energy, and Cascade tenants. This report focuses on the following three primary BA research questions: 1. What are the modeled energy savings using DOE low income weatherization approved TREAT software? 2. How did the modeled energy savings compare with measured energy savings from aggregate utility billing analysis? 3. What is the Savings to Investment Ratio (SIR) of the retrofit package after considering utility window incentives and KCHA capitol improvement funding.

  9. Regimes of turbulence without an energy cascade

    CERN Document Server

    Barenghi, C F; Baggaley, A W

    2016-01-01

    Experiments and numerical simulations of turbulent $^4$He and $^3$He-B have established that, at hydrodynamic length scales larger than the average distance between quantum vortices, the energy spectrum obeys the same 5/3 Kolmogorov law which is observed in the homogeneous isotropic turbulence of ordinary fluids. The importance of the 5/3 law is that it points to the existence of a Richardson energy cascade from large eddies to small eddies. However, there is also evidence of quantum turbulent regimes without Kolmogorov scaling. This raises the important questions of why, in such regimes, the Kolmogorov spectrum fails to form, what is the physical nature of turbulence without energy cascade, and whether hydrodynamical models can account for the unusual behaviour of turbulent superfluid helium. In this work we describe simple physical mechanisms which prevent the formation of Kolmogorov scaling in the thermal counterflow, and analyze the conditions necessary for emergence of quasiclassical regime in quantum tu...

  10. Cascaded Multicell Trans-Z-Source Inverters

    DEFF Research Database (Denmark)

    Li, Ding; Chiang Loh, Poh; Zhu, Miao

    2013-01-01

    strictly be small to prevent overvoltages caused by switching of their winding currents. To avoid these related problems, cascaded trans-Z-source inverters are proposed. They use multiple magnetic cells in an alternately cascading pattern rather than a single magnetic cell with large turns ratio....... Simulation and experimental results have shown that the multicell inverters can produce the same high-voltage gain, while keeping currents and voltages of the components low. The inverters can also step down their output voltages like a traditional voltage-source inverter without compromising waveform......Inverters with high-output voltage gain usually face the problem of high-input current flowing through their components. The problem might further be exaggerated if the inverters use high-frequency magnetic devices like transformers or coupled inductors. Leakage inductances of these devices must...

  11. Cascaded impedance networks for NPC inverter

    DEFF Research Database (Denmark)

    Li, Ding; Gao, Feng; Loh, Poh Chiang

    2010-01-01

    Multilevel inverters are getting more and more attracted because they have better output waveform quality and lower semiconductor voltage stress. The Z-source NPC is a kind of single stage multilevel inverter which has the ability of voltage boost. But the boost capability is relatively low when...... they are subject to the renewable sources. To date, three distinct types of impedance networks can be summarized for implementing a hybrid source impedance network, which can in principle be combined and cascaded before connected to a NPC inverter by proposed two ways. The resulting cascaded impedance network NPC...... would have a higher output voltage gain. It is anticipated that it would help the formed inverters find applications in photovoltaic and other renewable systems, where a high voltage gain is usually requested. Experimental testing has already been conducted and verifies the theory....

  12. Cascaded trans-z-source inverters

    DEFF Research Database (Denmark)

    Li, Ding; Loh, Poh Chiang; Zhu, Miao

    2011-01-01

    Z-source inverter is a recently proposed single-stage inverter with added voltage-boost capability for complementing the usual voltage-buck operation of a traditional voltage-source inverter. As long as the transformer element added in to the z-source concept, a trans-z-source inverter with one...... transformer and one capacitor is reported recently. This paper has adapted the cascaded concept into the trans-z-source and trans-quasi-z-source inverters to extend each to the cascaded topologies before combination is made with allowing more sources embedded which reduces the capacitor voltage and enhanced...... the compatibility for distributed sources. Unlike existing techniques, voltage stresses within the proposed inverters are better distributed among the passive components. Theoretical analysis for explaining these operating features has already been discussed before simulation were performed and an experimental...

  13. Supersonic Chordwise Bending Flutter in Cascades

    Science.gov (United States)

    1975-05-31

    bonding agent was injected via hypodermic needles after the blade tabs were properly inserted, The integrity and repeatability of the mounting of the indi...in conjunction with NASTRAN predictions and supersonic cascade aerodynamic computa- tions. Comparisons between theory and experiment are discussed. DD...explored by using the theory of .ieferences 3 and 4 to calculate the aero- dynamic damping for a typical chordwise blade deformation (Reference 6 and

  14. How do cilia organize signalling cascades?

    Science.gov (United States)

    Nachury, Maxence V

    2014-09-05

    Cilia and flagella are closely related centriole-nucleated protrusions of the cell with roles in motility and signal transduction. Two of the best-studied signalling pathways organized by cilia are the transduction cascade for the morphogen Hedgehog in vertebrates and the mating pathway that initiates gamete fusion in the unicellular green alga Chlamydomonas reinhardtii. What is the role of cilia in these signalling transduction cascades? In both Hedgehog and mating pathways, all signalling intermediates have been found to localize to cilia, and, for some signalling factors, ciliary localization is regulated by pathway activation. Given a concentration factor of three orders of magnitude provided by translocating a protein into the cilium, the compartment model proposes that cilia act as miniaturized reaction tubes bringing signalling factors and processing enzymes in close proximity. On the other hand, the scaffolding model views the intraflagellar transport machinery, whose primary function is to build cilia and flagella, as a molecular scaffold for the mating transduction cascade at the flagellar membrane. While these models may coexist, it is hoped that a precise understanding of the mechanisms that govern signalling inside cilia will provide a satisfying answer to the question 'how do cilia organize signalling?'. This review covers the evidence supporting each model of signalling and outlines future directions that may address which model applies in given biological settings. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

  15. Cascade laser applications: trends and challenges

    Science.gov (United States)

    d'Humières, B.; Margoto, Éric; Fazilleau, Yves

    2016-03-01

    When analyses need rapid measurements, cost effective monitoring and miniaturization, tunable semiconductor lasers can be very good sources. Indeed, applications like on-field environmental gas analysis or in-line industrial process control are becoming available thanks to the advantage of tunable semiconductor lasers. Advances in cascade lasers (CL) are revolutionizing Mid-IR spectroscopy with two alternatives: interband cascade lasers (ICL) in the 3-6μm spectrum and quantum cascade lasers (QCL), with more power from 3 to 300μm. The market is getting mature with strong players for driving applications like industry, environment, life science or transports. CL are not the only Mid-IR laser source. In fact, a strong competition is now taking place with other technologies like: OPO, VCSEL, Solid State lasers, Gas, SC Infrared or fiber lasers. In other words, CL have to conquer a share of the Mid-IR application market. Our study is a market analysis of CL technologies and their applications. It shows that improvements of components performance, along with the progress of infrared laser spectroscopy will drive the CL market growth. We compare CL technologies with other Mid-IR sources and estimate their share in each application market.

  16. Analysis of cascading failure in gene networks

    Directory of Open Access Journals (Sweden)

    Shudong eWang

    2012-12-01

    Full Text Available It is an important subject to research the functional mechanism of cancer-related genes make in formation and development of cancers. The modern methodology of data analysis plays a very important role for deducing the relationship between cancers and cancer-related genes and analyzing functional mechanism of genome. In this research, we construct mutual information networks using gene expression profiles of glioblast and renal in normal condition and cancer conditions. We investigate the relationship between structure and robustness in gene networks of the two tissues using a cascading failure model based on betweenness centrality. Define some important parameters such as the percentage of failure nodes of the network, the average size-ratio of cascading failure and the cumulative probability of size-ratio of cascading failure to measure the robustness of the networks. By comparing control group and experiment groups, we find that the networks of experiment groups are more robust than that of control group. The gene that can cause large scale failure is called structural key gene (SKG. Some of them have been confirmed to be closely related to the formation and development of glioma and renal cancer respectively. Most of them are predicted to play important roles during the formation of glioma and renal cancer, maybe the oncogenes, suppressor genes, and other cancer candidate genes in the glioma and renal cancer cells. However, these studies provide little information about the detailed roles of identified cancer genes.

  17. Streptococcal screen

    Science.gov (United States)

    ... Considerations This test screens for the group A streptococcus bacteria only. It ... MJ, eds. Mandell, Douglas and Bennett's Principles and Practice of Infectious Diseases . 8th ed. Philadelphia, PA: Elsevier Saunders; 2015:chap ...

  18. Hypertension screening

    Science.gov (United States)

    Foulke, J. M.

    1975-01-01

    An attempt was made to measure the response to an announcement of hypertension screening at the Goddard Space Center, to compare the results to those of previous statistics. Education and patient awareness of the problem were stressed.

  19. Lipid-cytokine-chemokine cascades orchestrate leukocyte recruitment in inflammation.

    Science.gov (United States)

    Sadik, Christian D; Luster, Andrew D

    2012-02-01

    Chemoattractants are pivotal mediators of host defense, orchestrating the recruitment of immune cells into sites of infection and inflammation. Chemoattractants display vast chemical diversity and include bioactive lipids, proteolytic fragments of serum proteins, and chemokines (chemotactic cytokines). All chemoattractants induce chemotaxis by activating seven-transmembrane-spanning GPCRs expressed on immune cells, establishing the concept that all chemoattractants are related in function. However, although chemoattractants have overlapping functions in vitro, recent in vivo data have revealed that they function, in many cases, nonredundantly in vivo. The chemically diverse nature of chemoattractants contributes to the fine control of leukocyte trafficking in vivo, with sequential chemoattractant use guiding immune cell recruitment into inflammatory sites. Lipid mediators frequently function as initiators of leukocyte recruitment, attracting the first immune cells into tissues. These initial responding immune cells produce cytokines locally, which in turn, induce the local release of chemokines. Local chemokine production then markedly amplifies subsequent waves of leukocyte recruitment. These new discoveries establish a paradigm for leukocyte recruitment in inflammation--described as lipid-cytokine-chemokine cascades--as a driving force in the effector phase of immune responses.

  20. HCC screening; HCC-Screening

    Energy Technology Data Exchange (ETDEWEB)

    Albrecht, T. [Charite-Unversitaetsmedizin,Freie Universitaet und Humboldt-Universitaet zu Berlin, Klinik und Hochschulambulanz fuer Radiologie und Nuklearmedizin,Campus Benjamin Franklin, Berlin (Germany)

    2008-01-15

    Hepatocellular carcinoma (HCC) is one of the most frequently diagnosed tumour diseases throughout the world. In the vast majority of cases those affected are high-risk patients with chronic viral hepatitis and/or liver cirrhosis, which means there is a clearly identifiable target group for HCC screening. With resection, transplantation, and interventional procedures for local ablation, following early diagnosis curative treatment options are available with which 5-year survival rates of over 60% can be reached. Such early diagnosis is a reality only in a minority of patients, however, and in the majority of cases the disease is already in an advanced stage at diagnosis. One of the objects of HCC screening is diagnosis in an early stage when curative treatment is still possible. Precisely this is achieved by screening, so that the proportion of patients treated with curative intent is decisively higher. There is not yet any clear evidence as to whether this leads to a lowering of the mortality of HCC. As lower mortality is the decisive indicator of success for a screening programme the benefit of HCC screening has so far been neither documented nor refuted. Nonetheless, in large regions of the world it is the practice for high-risk patients to undergo HCC screening in the form of twice-yearly ultrasound examination and determination of AFP. (orig.) [German] Das hepatozellulaere Karzinom (HCC) ist eine der weltweit haeufigsten Tumorerkrankungen. Es tritt in der grossen Mehrzahl der Faelle bei Hochrisikopatienten mit chronischer Virushepatitis bzw. Leberzirrhose auf, woraus sich eine klar identifizierbare Zielgruppe fuer das HCC-Screening ergibt. Mit der Resektion, der Transplantation und interventionellen lokal ablativen Verfahren stehen bei rechtzeitiger Diagnosestellung kurative Therapieoptionen zur Verfuegung, die 5-Jahres-Ueberlebensraten von >60% erreichen. Diese rechtzeitige Diagnosestellung erfolgt jedoch nur bei einer Minderzahl der Patienten, waehrend die

  1. Heat cascading regenerative sorption heat pump

    Science.gov (United States)

    Jones, Jack A. (Inventor)

    1995-01-01

    A simple heat cascading regenerative sorption heat pump process with rejected or waste heat from a higher temperature chemisorption circuit (HTCC) powering a lower temperature physisorption circuit (LTPC) which provides a 30% total improvement over simple regenerative physisorption compression heat pumps when ammonia is both the chemisorbate and physisorbate, and a total improvement of 50% or more for LTPC having two pressure stages. The HTCC contains ammonia and a chemisorbent therefor contained in a plurality of canisters, a condenser-evaporator-radiator system, and a heater, operatively connected together. The LTPC contains ammonia and a physisorbent therefor contained in a plurality of compressors, a condenser-evaporator-radiator system, operatively connected together. A closed heat transfer circuit (CHTC) is provided which contains a flowing heat transfer liquid (FHTL) in thermal communication with each canister and each compressor for cascading heat from the HTCC to the LTPC. Heat is regenerated within the LTPC by transferring heat from one compressor to another. In one embodiment the regeneration is performed by another CHTC containing another FHTL in thermal communication with each compressor. In another embodiment the HTCC powers a lower temperature ammonia water absorption circuit (LTAWAC) which contains a generator-absorber system containing the absorbent, and a condenser-evaporator-radiator system, operatively connected together. The absorbent is water or an absorbent aqueous solution. A CHTC is provided which contains a FHTL in thermal communication with the generator for cascading heat from the HTCC to the LTAWAC. Heat is regenerated within the LTAWAC by transferring heat from the generator to the absorber. The chemical composition of the chemisorbent is different than the chemical composition of the physisorbent, and the absorbent. The chemical composition of the FHTL is different than the chemisorbent, the physisorbent, the absorbent, and ammonia.

  2. A synthetic peptidoglycan fragment as a competitive inhibitor of the melanization cascade.

    Science.gov (United States)

    Park, Ji Won; Je, Byung-Rok; Piao, Shunfu; Inamura, Seiichi; Fujimoto, Yukari; Fukase, Koichi; Kusumoto, Shoichi; Söderhäll, Kenneth; Ha, Nam-Chul; Lee, Bok Luel

    2006-03-24

    Melanin synthesis is essential for defense and development but must be tightly controlled because systemic hyperactivation of the prophenoloxidase and excessive melanin synthesis are deleterious to the hosts. The melanization cascade of the arthropods can be activated by bacterial lysine-peptidoglycan (PGN), diaminopimelic acid (DAP)-PGN, or fungal beta-1,3-glucan. The molecular mechanism of how DAP- or Lys-PGN induces melanin synthesis and which molecules are involved in distinguishing these PGNs are not known. The identification of PGN derivatives that can work as inhibitors of the melanization cascade and the characterization of PGN recognition molecules will provide important information to clarify how the melanization is regulated and controlled. Here, we report that a novel synthetic Lys-PGN fragment ((GlcNAc-MurNAc-L-Ala-D-isoGln-L-Lys-D-Ala)2, T-4P2) functions as a competitive inhibitor of the natural PGN-induced melanization reaction. By using a T-4P2-coupled column, we purified the Tenebrio molitor PGN recognition protein (Tm-PGRP) without causing activation of the prophenoloxidase. The purified Tm-PGRP recognized both Lys- and DAP-PGN. In vitro reconstitution experiments showed that Tm-PGRP functions as a common recognition molecule of Lys- and DAP-PGN-dependent melanization cascades.

  3. The identification of a cascade hypernucleus

    CERN Document Server

    Mondal, A S; Husain, A; Kasim, M M

    1979-01-01

    In a systematic search for rare hypernuclear species in nuclear emulsion exposed to 3.0 GeV/c K/sup -/-mesons at the CERN PS, an event with three connecting stars has been observed. The two secondary stars are most probably due to the decay of a cascade hypernucleus according to the following channel: /sub Xi //sup -13/-C to /sub Lambda //sup 8 /Be+/sub Lambda //sup 5/He+Q. The binding energy of the Xi - hypernucleus is B/sub Xi /-(/sub Xi //sup 13/-C)=(18.1+or-3.2) MeV. (11 refs).

  4. Chronic traumatic encephalopathy and the availability cascade.

    Science.gov (United States)

    Solomon, Gary S; Sills, Allen

    2014-09-01

    Chronic traumatic encephalopathy (CTE) in sports has been known for > 85 years, and has experienced a resurgence of interest over the past decade, both in the media and in the scientific community. However, there appears to be a disconnection between the public's perception of CTE and the currently available scientific data. The cognitive bias known as the "availability cascade" has been suggested as a reason to explain this rift in knowledge. This review summarizes and updates the history of CTE in sports, discusses recent epidemiological and autopsy studies, summarizes the evidence base related to CTE in sports, and offers recommendations for future directions.

  5. The Cascade of Non-Stationarity

    Science.gov (United States)

    Belmont, P.; Kumarasamy, K.; Kelly, S. A.; Schaffrath, K. R.; Beach, T. J.

    2014-12-01

    Landscapes and channel networks are dynamic systems, often characterized by immense variability in time and space. Systematic shifts in hydrologic, geomorphic, or ecologic drivers can cause a cascade of changes within the system, which may fundamentally alter the way the system itself functions. Due to variability in resilience and resisting forces throughout the landscape, this cascade of changes may manifest in different ways within any given system. Humans may also exert considerable influence, often amplifying or damping system response. We illustrate the cascading effects of non-stationary hydrology and geomorphology in the Minnesota River Basin (MRB), a 44,000 km2 natural laboratory in which pervasive landscape disturbance has been triggered by several well-documented events. Rapid base-level lowering 13,400 YBP along the mainstem Minnesota River created a wave of incision, which continues to propagate up tributary channel networks. Temperature and precipitation have changed significantly in the MRB over the past century with rising temperatures, shifting precipitation patterns and an increase in heavy rainfall events. Streamflow has changed drastically and variably throughout the basin with 5% exceedance flows increasing 60-100% in recent decades, as increases in precipitation have been amplified by land management and artificial drainage. Increases in channel width and depth have occurred variably in the mainstem Minnesota River, the actively incising lower (knick zone) reaches of tributaries, and the low gradient, passively meandering reaches above the knick zones. Altered hydrologic regimes and channel morphologies, combined with increased sedimentation and nutrient loading have adversely affected aquatic biota via disruption of life cycles and habitat degradation. Existing landscape, water quality, and flood risk models are poorly equipped to deal with the cascading effects of non-stationarity and therefore may grossly over- or under

  6. Tracking Control for Switched Cascade Nonlinear Systems

    Directory of Open Access Journals (Sweden)

    Xiaoxiao Dong

    2015-01-01

    Full Text Available The issue of H∞ output tracking for switched cascade nonlinear systems is discussed in this paper, where not all the linear parts of subsystems are stabilizable. The conditions of the solvability for the issue are given by virtue of the structural characteristics of the systems and the average dwell time method, in which the total activation time for stabilizable subsystems is longer than that for the unstabilizable subsystems. At last, a simulation example is used to demonstrate the validity and advantages of the proposed approach.

  7. Cascaded recompression closed brayton cycle system

    Energy Technology Data Exchange (ETDEWEB)

    Pasch, James J.

    2018-01-02

    The present disclosure is directed to a cascaded recompression closed Brayton cycle (CRCBC) system and method of operation thereof, where the CRCBC system includes a compressor for compressing the system fluid, a separator for generating fluid feed streams for each of the system's turbines, and separate segments of a heater that heat the fluid feed streams to different feed temperatures for the system's turbines. Fluid exiting each turbine is used to preheat the fluid to the turbine. In an embodiment, the amount of heat extracted is determined by operational costs.

  8. Long-Haul TCP vs. Cascaded TCP

    OpenAIRE

    Feng, Wu-chun

    2006-01-01

    In this work, we investigate the bandwidth and transfer time of long-haul TCP versus cascaded TCP [5]. First, we discuss the models for TCP throughput. For TCP flows in support of bulk data transfer (i.e., long-lived TCP flows), the TCP throughput models have been derived [2, 3]. These models rely on the congestion-avoidance algorithm of TCP. Though these models cannot be applied with short-lived TCP connections, our interest relative to logistical networking is in longer-li...

  9. Plant MAPK cascades: Just rapid signaling modules?

    KAUST Repository

    Boudsocq, Marie

    2015-08-27

    © 2015 Taylor & Francis Group, LLC. Abscisic acid (ABA) is a major phytohormone mediating important stress-related processes. We recently unveiled an ABA-activated MAPK signaling module constituted of MAP3K17/18-MKK3-MPK1/2/7/14. Unlike classical rapid MAPK activation, we showed that the activation of the new MAPK module is delayed and relies on the MAP3K protein synthesis. In this addendum, we discuss the role of this original and unexpected activation mechanism of MAPK cascades which suggests that MAPKs can regulate both early and longterm plant stress responses.

  10. The impact of a network split on cascading failure processes

    OpenAIRE

    Sloothaak, Fiona; Borst, Sem C.; Zwart, Bert

    2017-01-01

    Cascading failure models are typically used to capture the phenomenon where failures possibly trigger further failures in succession, causing knock-on effects. In many networks this ultimately leads to a disintegrated network where the failure propagation continues independently across the various components. In order to gain insight in the impact of network splitting on cascading failure processes, we extend a well-established cascading failure model for which the number of failures obeys a ...

  11. Large phase shift via polarization-coupling cascading.

    Science.gov (United States)

    Huo, Juan; Chen, Xianfeng

    2012-06-04

    Herein, we propose a phenomenon of "polarization-coupling (PC) cascading" generated in MgO doped periodically poled lithium niobate crystal (PPMgLN). PC cascading contributes to the effective electro-optical (EO) Kerr effect that is several orders of magnitude stronger than the classical ones. Experiment of Newton's rings demonstrates the large phase accumulation during the PC cascaded processes, and the experimental data is identical with the theoretical simulation.

  12. Estimating Failure Propagation in Models of Cascading Blackouts

    Energy Technology Data Exchange (ETDEWEB)

    Dobson, Ian [University of Wisconsin, Madison; Carreras, Benjamin A [ORNL; Lynch, Vickie E [ORNL; Nkei, Bertrand [ORNL; Newman, David E [University of Alaska

    2005-09-01

    We compare and test statistical estimates of failure propagation in data from versions of a probabilistic model of loading-dependent cascading failure and a power systems blackout model of cascading transmission line overloads. The comparisons suggest mechanisms affecting failure propagation and are an initial step towards monitoring failure propagation from practical system data. Approximations to the probabilistic model describe the forms of probability distributions of cascade sizes.

  13. 21 CFR 866.2420 - Oxidase screening test for gonorrhea.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Oxidase screening test for gonorrhea. 866.2420... screening test for gonorrhea. (a) Identification. An oxidase screening test for gonorrhea is an in vitro... of gonorrhea. (b) Classification. Class III (premarket approval) (transitional device). (c) Date PMA...

  14. All passive architecture for high efficiency cascaded Raman conversion

    Science.gov (United States)

    Balaswamy, V.; Arun, S.; Chayran, G.; Supradeepa, V. R.

    2018-02-01

    Cascaded Raman fiber lasers have offered a convenient method to obtain scalable, high-power sources at various wavelength regions inaccessible with rare-earth doped fiber lasers. A limitation previously was the reduced efficiency of these lasers. Recently, new architectures have been proposed to enhance efficiency, but this came at the cost of enhanced complexity, requiring an additional low-power, cascaded Raman laser. In this work, we overcome this with a new, all-passive architecture for high-efficiency cascaded Raman conversion. We demonstrate our architecture with a fifth-order cascaded Raman converter from 1117nm to 1480nm with output power of ~64W and efficiency of 60%.

  15. Cascading walks model for human mobility patterns.

    Science.gov (United States)

    Han, Xiao-Pu; Wang, Xiang-Wen; Yan, Xiao-Yong; Wang, Bing-Hong

    2015-01-01

    Uncovering the mechanism behind the scaling laws and series of anomalies in human trajectories is of fundamental significance in understanding many spatio-temporal phenomena. Recently, several models, e.g. the explorations-returns model (Song et al., 2010) and the radiation model for intercity travels (Simini et al., 2012), have been proposed to study the origin of these anomalies and the prediction of human movements. However, an agent-based model that could reproduce most of empirical observations without priori is still lacking. In this paper, considering the empirical findings on the correlations of move-lengths and staying time in human trips, we propose a simple model which is mainly based on the cascading processes to capture the human mobility patterns. In this model, each long-range movement activates series of shorter movements that are organized by the law of localized explorations and preferential returns in prescribed region. Based on the numerical simulations and analytical studies, we show more than five statistical characters that are well consistent with the empirical observations, including several types of scaling anomalies and the ultraslow diffusion properties, implying the cascading processes associated with the localized exploration and preferential returns are indeed a key in the understanding of human mobility activities. Moreover, the model shows both of the diverse individual mobility and aggregated scaling displacements, bridging the micro and macro patterns in human mobility. In summary, our model successfully explains most of empirical findings and provides deeper understandings on the emergence of human mobility patterns.

  16. Quaternary Magmatism in the Cascades - Geologic Perspectives

    Science.gov (United States)

    Hildreth, Wes

    2007-01-01

    Foreward The Cascade magmatic arc is a belt of Quaternary volcanoes that extends 1,250 km from Lassen Peak in northern California to Meager Mountain in Canada, above the subduction zone where the Juan de Fuca Plate plunges beneath the North American Plate. This Professional Paper presents a synthesis of the entire volcanic arc, addressing all 2,300 known Quaternary volcanoes, not just the 30 or so visually prominent peaks that comprise the volcanic skyline. Study of Cascade volcanoes goes back to the geological explorers of the late 19th century and the seminal investigations of Howel Williams in the 1920s and 1930s. However, major progress and application of modern scientific methods and instrumentation began only in the 1970s with the advent of systematic geological, geophysical, and geochemical studies of the entire arc. Initial stimulus from the USGS Geothermal Research Program was enhanced by the USGS Volcano Hazards Program following the 1980 eruption of Mount St. Helens. Together, these two USGS Programs have provided more than three decades of stable funding, staffing, and analytical support. This Professional Paper summarizes the resultant USGS data sets and integrates them with the parallel contributions of other investigators. The product is based upon an all-encompassing and definitive geological database, including chemical and isotopic analyses to characterize the rocks and geochronology to provide the critical time constraints. Until now, this massive amount of data has not been summarized, and a systematic and uniform interpretation firmly grounded in geological fact has been lacking. Herein lies the primary utility of this Cascade volume. It not only will be the mandatory starting point for new workers, but also will provide essential geological context to broaden the perspectives of current investigators of specific Cascade volcanoes. Wes Hildreth's insightful understanding of volcanic processes and his uncompromising scientific integrity make him

  17. Testicular Cancer Screening

    Science.gov (United States)

    ... Professional Testicular Cancer Treatment Testicular Cancer Screening Testicular Cancer Screening (PDQ®)–Patient Version What is screening? Go to ... testicles, and need to be followed closely. Testicular Cancer Screening Key Points Tests are used to screen for ...

  18. Alcohol Use Screening

    Science.gov (United States)

    ... Depression Screening Substance Abuse Screening Alcohol Use Screening Alcohol Use Screening (AUDIT-C) - Instructions The following questions ... this tool, there is also text-only version . Alcohol Use Screening (AUDIT-C) - Manual Instructions The following ...

  19. Hearing Screening

    Science.gov (United States)

    Johnson-Curiskis, Nanette

    2012-01-01

    Hearing levels are threatened by modern life--headsets for music, rock concerts, traffic noises, etc. It is crucial we know our hearing levels so that we can draw attention to potential problems. This exercise requires that students receive a hearing screening for their benefit as well as for making the connection of hearing to listening.

  20. Screening Tests

    Science.gov (United States)

    ... gov/publications/AssessingAlcohol/index.htm .) This issue of Alcohol Research & Health highlights some of the most popular screening ... tolerance to more than two drinks (the T question) = 2 points. The Alcohol Use Disorders Identification Test (AUDIT) can detect alcohol ...

  1. Eruptive history of South Sister, Oregon Cascades

    Science.gov (United States)

    Fierstein, J.; Hildreth, W.; Calvert, A.T.

    2011-01-01

    South Sister is southernmost and highest of the Three Sisters, three geologically dissimilar stratovolcanoes that together form a spectacular 20km reach along the Cascade crest in Oregon. North Sister is a monotonously mafic edifice as old as middle Pleistocene, Middle Sister a basalt-andesite-dacite cone built between 48 and 14ka, and South Sister is a basalt-free edifice that alternated rhyolitic and intermediate modes from 50ka to 2ka (largely contemporaneous with Middle Sister). Detailed mapping, 330 chemical analyses, and 42 radioisotopic ages show that the oldest exposed South Sister lavas were initially rhyolitic ~50ka. By ~37ka, rhyolitic lava flows and domes (72-74% SiO2) began alternating with radially emplaced dacite (63-68% SiO2) and andesite (59-63% SiO2) lava flows. Construction of a broad cone of silicic andesite-dacite (61-64% SiO2) culminated ~30ka in a dominantly explosive sequence that began with crater-forming andesitic eruptions that left fragmental deposits at least 200m thick. This was followed at ~27ka by growth of a steeply dipping summit cone of agglutinate-dominated andesite (56-60.5% SiO2) and formation of a summit crater ~800m wide. This crater was soon filled and overtopped by a thick dacite lava flow and then by >150m of dacitic pyroclastic ejecta. Small-volume dacite lavas (63-67% SiO2) locally cap the pyroclastic pile. A final sheet of mafic agglutinate (54-56% SiO2) - the most mafic product of South Sister - erupted from and drapes the small (300-m-wide) present-day summit crater, ending a summit-building sequence that lasted until ~22ka. A 20kyr-long-hiatus was broken by rhyolite eruptions that produced (1) the Rock Mesa coulee, tephra, and satellite domelets (73.5% SiO2) and (2) the Devils Chain of ~20 domes and short coulees (72.3-72.8% SiO2) from N-S vent alignments on South Sister's flanks. The compositional reversal from mafic summit agglutinate to recent rhyolites epitomizes the frequently changing compositional modes of the

  2. Temporal switching induced by cascaded third order nonlinearity

    DEFF Research Database (Denmark)

    Eilenberger, Falk; Bache, Morten; Minardi, Stefano

    2012-01-01

    We investigate the impact of cascaded third harmonic generation and the intrinsic n4 material nonlinearity on the propagation of ultrashort pulses in noble-gas filled Kagome fibers. We show that the pressure tunability of the cascade allows for the implementation of temporal switching. We also...... investigate the relative strengths of both effects and show their ratio to be pressure tunable....

  3. Cascade vulnerability for risk analysis of water infrastructure.

    Science.gov (United States)

    Sitzenfrei, R; Mair, M; Möderl, M; Rauch, W

    2011-01-01

    One of the major tasks in urban water management is failure-free operation for at least most of the time. Accordingly, the reliability of the network systems in urban water management has a crucial role. The failure of a component in these systems impacts potable water distribution and urban drainage. Therefore, water distribution and urban drainage systems are categorized as critical infrastructure. Vulnerability is the degree to which a system is likely to experience harm induced by perturbation or stress. However, for risk assessment, we usually assume that events and failures are singular and independent, i.e. several simultaneous events and cascading events are unconsidered. Although failures can be causally linked, a simultaneous consideration in risk analysis is hardly considered. To close this gap, this work introduces the term cascade vulnerability for water infrastructure. Cascade vulnerability accounts for cascading and simultaneous events. Following this definition, cascade risk maps are a merger of hazard and cascade vulnerability maps. In this work cascade vulnerability maps for water distribution systems and urban drainage systems based on the 'Achilles-Approach' are introduced and discussed. It is shown, that neglecting cascading effects results in significant underestimation of risk scenarios.

  4. A nine-level hybrid symmetric cascaded multilevel converter for ...

    Indian Academy of Sciences (India)

    Bridge (TCHB) power cell and one three-level H-bridge power cell with equal dc link voltages, and they are connected in cascade. Due to cascade connection and equal dc link voltage, the power shared by each power cell is nearly equal.

  5. Hybrid Modulation Scheme for Cascaded H-Bridge Inverter Cells ...

    African Journals Online (AJOL)

    This work proposes a switching technique for cascaded H-Bridge (CHB) cells. Single carrier Sinusoidal PWM (SCSPWM) scheme is employed in the generation of the gating signals. A sequential switching and base PWM circulation schemes are presented for this fundamental cascaded multilevel inverter topology.

  6. Influence of blood flow on the coagulation cascade

    DEFF Research Database (Denmark)

    The influence of diffusion and convetive flows on the blood coagulation cascade is investigated for a controlled perfusion experiment. We present a cartoon model and reaction schemes for parts of the coagulation cascade with sunsequent set up of a mathematical model in two space dimensions plus one...

  7. Seeking the northern boundary of the Cascade strawberry

    Science.gov (United States)

    Fragaria cascadensis K.E. Hummer, an endemic decaploid strawberry species, was described from the Oregon Cascade Mountains in the Pacific North-western United States. Its range occurs near Mt. Hood, the highest peak in the northern Oregon Cascades, in a band of higher elevation sites southwards to n...

  8. 36 CFR 7.66 - North Cascades National Park.

    Science.gov (United States)

    2010-07-01

    ... 36 Parks, Forests, and Public Property 1 2010-07-01 2010-07-01 false North Cascades National Park. 7.66 Section 7.66 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR SPECIAL REGULATIONS, AREAS OF THE NATIONAL PARK SYSTEM § 7.66 North Cascades National Park. (a...

  9. Intermittent Flow Regimes in a Transonic Fan Airfoil Cascade

    Directory of Open Access Journals (Sweden)

    J. Lepicovsky

    2004-01-01

    velocity.To date, this flow behavior has only been observed in a linear transonic cascade. Further research is necessary to confirm this phenomenon occurs in actual transonic fans and is not the by-product of an endwall restricted linear cascade.

  10. NIR-triggered high-efficient photodynamic and chemo-cascade therapy using caspase-3 responsive functionalized upconversion nanoparticles.

    Science.gov (United States)

    Zhao, Na; Wu, Baoyan; Hu, Xianglong; Xing, Da

    2017-10-01

    Stimuli-responsive nanoparticles with multiple therapeutic/diagnostic functions are highly desirable for effective tumor treatment. Herein novel caspase-3 responsive functionalized upconversion nanoparticles (CFUNs) were fabricated with three-in-one functional integration: near-infrared (NIR) triggered photodynamic damage along with caspase-3 activation, subsequent caspase-3 responsive drug release, and cascade chemotherapeutic activation. CFUNs were formulated from the self-assembly of caspase-3 responsive doxorubicin (DOX) prodrug tethered with DEVD peptide (DEVD-DOX), upconversion nanoparticles (UCNP), a photosensitizer (pyropheophorbide-a methyl ester, MPPa), and tumor-targeting cRGD-PEG-DSPE to afford multifunctional CFUNs, MPPa/UCNP-DEVD-DOX/cRGD. Upon cellular uptake and NIR irradiation, the visible light emission of UCNP could excite MPPa to produce reactive oxygen species for photodynamic therapy (PDT) along with the activation of caspase-3, which further cleaved DEVD peptide to release DOX within tumor cells, thus accomplishing NIR-triggered PDT and cascade chemotherapy. CFUNs presented silent therapeutic potency and negligible cytotoxicity in the dark, whereas in vitro and in vivo experiments demonstrated the NIR-triggered cascade therapeutic activation and tumor inhibition due to consecutive PDT and chemotherapy. Current NIR-activated cascade tumor therapy with two distinct mechanisms is significantly favorable to overcome multidrug resistance and tumor heterogeneity for persistent tumor treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. A modeling framework for system restoration from cascading failures.

    Science.gov (United States)

    Liu, Chaoran; Li, Daqing; Zio, Enrico; Kang, Rui

    2014-01-01

    System restoration from cascading failures is an integral part of the overall defense against catastrophic breakdown in networked critical infrastructures. From the outbreak of cascading failures to the system complete breakdown, actions can be taken to prevent failure propagation through the entire network. While most analysis efforts have been carried out before or after cascading failures, restoration during cascading failures has been rarely studied. In this paper, we present a modeling framework to investigate the effects of in-process restoration, which depends strongly on the timing and strength of the restoration actions. Furthermore, in the model we also consider additional disturbances to the system due to restoration actions themselves. We demonstrate that the effect of restoration is also influenced by the combination of system loading level and restoration disturbance. Our modeling framework will help to provide insights on practical restoration from cascading failures and guide improvements of reliability and resilience of actual network systems.

  12. Impedance Coordinative Control for Cascaded Converter in Bidirectional Application

    DEFF Research Database (Denmark)

    Tian, Yanjun; Deng, Fujin; Chen, Zhe

    2015-01-01

    Two stage cascaded converters are widely used in bidirectional applications, but the negative impedance may cause system instability. Actually the impedance interaction is much different between forward power flow and reversed power flow, which will introduce more uncertainty to the system...... stability. This paper proposes a control method for the constant power controlled converter in cascaded system, and consequently it can change the negative impedance of constant power converter into resistive impedance, which will improve the cascaded system stability, as well as merge the impedance...... difference between forward and reversed power flow. This paper addresses the analysis with the topology of cascaded dual-active-bridge converter (DAB) with inverter, and the proposed control method can also be implemented in unidirectional applications and other general cascaded converter system...

  13. A modeling framework for system restoration from cascading failures.

    Directory of Open Access Journals (Sweden)

    Chaoran Liu

    Full Text Available System restoration from cascading failures is an integral part of the overall defense against catastrophic breakdown in networked critical infrastructures. From the outbreak of cascading failures to the system complete breakdown, actions can be taken to prevent failure propagation through the entire network. While most analysis efforts have been carried out before or after cascading failures, restoration during cascading failures has been rarely studied. In this paper, we present a modeling framework to investigate the effects of in-process restoration, which depends strongly on the timing and strength of the restoration actions. Furthermore, in the model we also consider additional disturbances to the system due to restoration actions themselves. We demonstrate that the effect of restoration is also influenced by the combination of system loading level and restoration disturbance. Our modeling framework will help to provide insights on practical restoration from cascading failures and guide improvements of reliability and resilience of actual network systems.

  14. Modelling of Attenuation and Crosstalk of Cascaded Transmission Lines

    Directory of Open Access Journals (Sweden)

    Pavel Lafata

    2011-01-01

    Full Text Available This paper deals with the measurements and modelling of attenuation and near-end (NEXT and far-end (FEXT crosstalk for cascaded metallic transmission lines. The transmission parameters of homogenous metallic line can be easily described by telegraph equations or cascade matrix; there are also several models for NEXT and FEXT frequency dependence. But these models and equations could not be applied in the situation of two or more different cascaded transmission lines, because these cascaded lines do not meet the essential condition of overall homogenous transmission line. However in such case, it is still possible to estimate the overall transmission characteristics of the whole combination thanks to the characteristics of each separate element. This paper brings the description of complex measurements performed for the combination of three different metallic cables and based on these measurements, several conclusions about the possibilities of modelling the attenuation and NEXT and FEXT crosstalk for cascaded transmission lines are presented.

  15. Quantum cascade laser FM spectroscopy of explosives

    Science.gov (United States)

    Gutmann, Zach; Clasp, Trocia; Lue, Chris; Johnson, Tiffani; Ingle, Taylor; Jamison, Janet; Buchanan, Roger; Reeve, Scott

    2013-05-01

    Polyisobutylene is an industrial polymer that is widely used in a number of applications including the manufacture of military grade explosives. We have examined the vapor emanating from a series of different molecular weight samples of polyisobutylene using high resolution Quantum Cascade Laser FM spectroscopy. The vapor phase spectra all exhibit a rovibrational structure similar to that for the gas phase isobutylene molecule. We have assigned the structure in the 890 cm-1 and 1380 cm-1 regions to the isobutylene ν28 and ν7 fundamental bands respectively. These spectroscopic signatures may prove useful for infrared sensing applications. Here we will present the infrared signatures along with recent GCMS data from a sample of C4, utilizing solid-phase microextraction vapor collection fibers, which confirm the presence of isobutylene as one of the volatile bouquet species in RDX-based explosives.

  16. Control of cascaded induction generator systems

    Science.gov (United States)

    Ortmeyer, T. H.

    1984-12-01

    This report documents an investigation of the stability and control of cascaded doubly fed machines (CDFM). These machines are brushless variable speed constant frequency electric power generators with potential for application in aircraft. A previous analytical study indicated the CDFM system would be controllable in the subsynchronous operating mode with a passive RL load. The present study contains two steps. First is an investigation of the machine operation in the supersynchronous mode. The second step is an investigation of machine operation with output capacitors providing excitation VARs for the machine and load. Step 1 results show that the machines exhibit stability characteristics in the supersynchronous mode similar to those observed in the subsynchronous mode. Step 2 results show that output capacitors degrade the system performance, particularly at light loads. The results show that output current feedback can be employed to improve the system performance.

  17. Cascaded face alignment via intimacy definition feature

    Science.gov (United States)

    Li, Hailiang; Lam, Kin-Man; Chiu, Man-Yau; Wu, Kangheng; Lei, Zhibin

    2017-09-01

    Recent years have witnessed the emerging popularity of regression-based face aligners, which directly learn mappings between facial appearance and shape-increment manifolds. We propose a random-forest based, cascaded regression model for face alignment by using a locally lightweight feature, namely intimacy definition feature. This feature is more discriminative than the pose-indexed feature, more efficient than the histogram of oriented gradients feature and the scale-invariant feature transform feature, and more compact than the local binary feature (LBF). Experimental validation of our algorithm shows that our approach achieves state-of-the-art performance when testing on some challenging datasets. Compared with the LBF-based algorithm, our method achieves about twice the speed, 20% improvement in terms of alignment accuracy and saves an order of magnitude on memory requirement.

  18. Cascaded Amplitude Modulations in Sound Texture Perception

    DEFF Research Database (Denmark)

    McWalter, Richard Ian; Dau, Torsten

    2017-01-01

    Sound textures, such as crackling fire or chirping crickets, represent a broad class of sounds defined by their homogeneous temporal structure. It has been suggested that the perception of texture is mediated by time-averaged summary statistics measured from early auditory representations....... In this study, we investigated the perception of sound textures that contain rhythmic structure, specifically second-order amplitude modulations that arise from the interaction of different modulation rates, previously described as "beating" in the envelope-frequency domain. We developed an auditory texture...... model that utilizes a cascade of modulation filterbanks that capture the structure of simple rhythmic patterns. The model was examined in a series of psychophysical listening experiments using synthetic sound textures-stimuli generated using time-averaged statistics measured from real-world textures...

  19. Cascaded Quadratic Soliton Compression in Waveguide Structures

    DEFF Research Database (Denmark)

    Guo, Hairun

    between the Kerr nonlinear effects and the dispersive effects in the medium. A Kerr-like nonlinearity is produced through the cascaded phase mismatched quadratic process, e.g. the second harmonic generation process, which can be flexibly tuned in both the sign and the amplitude, making possible a strong...... and self-defocusing Kerr effect so that the soliton is created and the soliton self-compression happens in the normal dispersion region. Meanwhile, the chromatic dispersion in the waveguide is also tunable, understood as the dispersion engineering with structural designs. Therefore, compared to commonly......-focusing Kerr effects when under the self-defocusing regime. On the other hand, CQSC in quadratic waveguides seems highly complementary to that in quadratic bulk crystals. With bulk crystals dealing with high-energy, low-repetition-rate and large-beam-size pulses, quadratic waveguides could operate low...

  20. Cascaded FSO-VLC Communication System

    KAUST Repository

    Gupta, Akash

    2017-08-28

    The proposed cascaded free space optics (FSO)-visible light communication (VLC) system consists of multiple VLC access points which caters the end users connected via a decode and forward (DF) relay to the FSO backhaul link. The FSO link is assumed to be affected by path-loss, pointing error and atmospheric turbulence while the end-to-end signal-to-noise ratio (SNR) of VLC downlinks are statistically characterized considering the randomness of users position. In this study, the novel closed form expressions of the statistics like probability density function (PDF) and cumulative distribution function (CDF) of the equivalent SNR are derived. Capitalizing on these, the closed form expressions for various performance metrics such as outage probability and error probability are provided. The simulation results are provided to verify the functional curves of mathematical analysis.

  1. Information propagation in a noisy gene cascade

    Science.gov (United States)

    Monteoliva, D.; Diambra, L.

    2017-07-01

    We use information theory to study the information transmission through a simple gene cascade where the product of an unregulated gene regulates the expression activity of a cooperative genetic switch. While the input signal is provided by the upstream gene with two states, we consider that the expression of downstream gene is controlled by a cis-regulatory system with three binding sites for the regulator product, which can bind cooperatively. By computing exactly the associated probability distributions, we estimate information transmission thought the mutual information measure. We found that the mutual information associated with unimodal input signal is lower than the associated with bimodal inputs. We also observe that mutual information presents a maximum in the cooperativity intensity, and the position of this maximum depends on the kinetic rates of the promoter. Furthermore, we found that the bursting dynamics of the input signal can enhance the information transmission capacity.

  2. Fluctuation sensitivity of a transcriptional signaling cascade

    Science.gov (United States)

    Pilkiewicz, Kevin R.; Mayo, Michael L.

    2016-09-01

    The internal biochemical state of a cell is regulated by a vast transcriptional network that kinetically correlates the concentrations of numerous proteins. Fluctuations in protein concentration that encode crucial information about this changing state must compete with fluctuations caused by the noisy cellular environment in order to successfully transmit information across the network. Oftentimes, one protein must regulate another through a sequence of intermediaries, and conventional wisdom, derived from the data processing inequality of information theory, leads us to expect that longer sequences should lose more information to noise. Using the metric of mutual information to characterize the fluctuation sensitivity of transcriptional signaling cascades, we find, counter to this expectation, that longer chains of regulatory interactions can instead lead to enhanced informational efficiency. We derive an analytic expression for the mutual information from a generalized chemical kinetics model that we reduce to simple, mass-action kinetics by linearizing for small fluctuations about the basal biological steady state, and we find that at long times this expression depends only on a simple ratio of protein production to destruction rates and the length of the cascade. We place bounds on the values of these parameters by requiring that the mutual information be at least one bit—otherwise, any received signal would be indistinguishable from noise—and we find not only that nature has devised a way to circumvent the data processing inequality, but that it must be circumvented to attain this one-bit threshold. We demonstrate how this result places informational and biochemical efficiency at odds with one another by correlating high transcription factor binding affinities with low informational output, and we conclude with an analysis of the validity of our assumptions and propose how they might be tested experimentally.

  3. Cascading walks model for human mobility patterns.

    Directory of Open Access Journals (Sweden)

    Xiao-Pu Han

    Full Text Available Uncovering the mechanism behind the scaling laws and series of anomalies in human trajectories is of fundamental significance in understanding many spatio-temporal phenomena. Recently, several models, e.g. the explorations-returns model (Song et al., 2010 and the radiation model for intercity travels (Simini et al., 2012, have been proposed to study the origin of these anomalies and the prediction of human movements. However, an agent-based model that could reproduce most of empirical observations without priori is still lacking.In this paper, considering the empirical findings on the correlations of move-lengths and staying time in human trips, we propose a simple model which is mainly based on the cascading processes to capture the human mobility patterns. In this model, each long-range movement activates series of shorter movements that are organized by the law of localized explorations and preferential returns in prescribed region.Based on the numerical simulations and analytical studies, we show more than five statistical characters that are well consistent with the empirical observations, including several types of scaling anomalies and the ultraslow diffusion properties, implying the cascading processes associated with the localized exploration and preferential returns are indeed a key in the understanding of human mobility activities. Moreover, the model shows both of the diverse individual mobility and aggregated scaling displacements, bridging the micro and macro patterns in human mobility. In summary, our model successfully explains most of empirical findings and provides deeper understandings on the emergence of human mobility patterns.

  4. Orexin/hypocretin receptor signalling cascades.

    Science.gov (United States)

    Kukkonen, J P; Leonard, C S

    2014-01-01

    Orexin (hypocretin) peptides and their two known G-protein-coupled receptors play essential roles in sleep-wake control and powerfully influence other systems regulating appetite/metabolism, stress and reward. Consequently, drugs that influence signalling by these receptors may provide novel therapeutic opportunities for treating sleep disorders, obesity and addiction. It is therefore critical to understand how these receptors operate, the nature of the signalling cascades they engage and their physiological targets. In this review, we evaluate what is currently known about orexin receptor signalling cascades, while a sister review (Leonard & Kukkonen, this issue) focuses on tissue-specific responses. The evidence suggests that orexin receptor signalling is multifaceted and is substantially more diverse than originally thought. Indeed, orexin receptors are able to couple to members of at least three G-protein families and possibly other proteins, through which they regulate non-selective cation channels, phospholipases, adenylyl cyclase, and protein and lipid kinases. In the central nervous system, orexin receptors produce neuroexcitation by postsynaptic depolarization via activation of non-selective cation channels, inhibition of K⁺ channels and activation of Na⁺/Ca²⁺ exchange, but they also can stimulate the release of neurotransmitters by presynaptic actions and modulate synaptic plasticity. Ca²⁺ signalling is also prominently influenced by these receptors, both via the classical phospholipase C-Ca²⁺ release pathway and via Ca²⁺ influx, mediated by several pathways. Upon longer-lasting stimulation, plastic effects are observed in some cell types, while others, especially cancer cells, are stimulated to die. Thus, orexin receptor signals appear highly tunable, depending on the milieu in which they are operating. © 2013 The British Pharmacological Society.

  5. Antifungal response of or-al-associated candidal reference strains (American Type Culture Collection by supercritical fluid extract of nutmeg seeds for geriatric denture wearers: An in vitro screening study

    Directory of Open Access Journals (Sweden)

    Meenakshi Iyer

    2017-01-01

    Conclusion: This paper described the in vitro antibacterial activity, and phytochemical analysis of SFE extract of nutmeg (Myristica fragrans evaluated against C. albicans (American Type Culture Collection 10231 through agar well diffusion method. SFE of nutmeg seeds can be used as an adjunct to conventional therapy for oral candidiasis.

  6. Allergic sensitization: screening methods

    DEFF Research Database (Denmark)

    Ladics, Gregory S.; Fry, Jeremy; Goodman, Richard

    2014-01-01

    Experimental in silico, in vitro, and rodent models for screening and predicting protein sensitizing potential are discussed, including whether there is evidence of new sensitizations and allergies since the introduction of genetically modified crops in 1996, the importance of linear versus...... conformational epitopes, and protein families that become allergens. Some common challenges for predicting protein sensitization are addressed: (a) exposure routes; (b) frequency and dose of exposure; (c) dose-response relationships; (d) role of digestion, food processing, and the food matrix; (e) role...... potential of a novel protein. However, they would be extremely useful in the discovery and research phases of understanding the mechanisms of food allergy development, and may prove fruitful to provide information regarding potential allergenicity risk assessment of future products on a case by case basis...

  7. Normalized-Mutual-Information-Based Mining Method for Cascading Patterns

    Directory of Open Access Journals (Sweden)

    Cunjin Xue

    2016-09-01

    Full Text Available A cascading pattern is a sequential pattern characterized by an item following another item in order. Recent research has investigated a challenge of dealing with cascading patterns, namely, the exponential time dependence of database scanning with respect to the number of items involved. We propose a normalized-mutual-information-based mining method for cascading patterns (M3Cap to address this challenge. M3Cap embeds mutual information to reduce database-scanning time. First, M3Cap calculates the asymmetrical mutual information between items with one database scan and extracts pair-wise related items according to a user-specified information threshold. Second, a one-level cascading pattern is generated by scanning the database once for each pair-wise related item at the quantitative level. Third, a recursive linking–pruning–generating loop generates an (m + 1-level-candidate cascading pattern from m-dimensional patterns on the basis of antimonotonicity and non-additivity, repeating this step until no further candidate cascading patterns are generated. Fourth, meaningful cascading patterns are generated according to user-specified minimum evaluation indicators. Finally, experiments with remote sensing image datasets covering the Pacific Ocean demonstrate that the computation time of recursive linking and pruning is significantly less than that of database scanning; thus, M3Cap improves performance by reducing database scanning while increasing intensive computing.

  8. Out of control: Fluctuation of cascading dynamics in networks

    Science.gov (United States)

    Wang, Jianwei; Cai, Lin; Xu, Bo; Li, Peng; Sun, Enhui; Zhu, Zhiguo

    2016-11-01

    Applying two preferential selection mechanisms of flow destination, we develop two new methods to quantify the initial load of a node, where the flow is transported along the shortest path between two nodes. We propose a simple cascading model and study cascading dynamics induced by attacking the node with the highest load in some synthetic and actual networks. Surprisingly, we observe the abnormal fluctuation of cascading dynamics, i.e., more damage can be triggered if we spend significantly higher cost to protect a network. In particular, this phenomenon is independent of the initial flow distribution and the preferential selection mechanisms of flow destination. However, it remains unclear which specific structural patterns may affect the fluctuation of cascading dynamics. In this paper, we examine the local evolution of the cascading propagation by constructing some special networks. We show that revivals of some nodes in the double ring structure facilitate the transportation of the flow between two unconnected sub-networks, cause more damage and subsequently lead to the abnormal fluctuation of cascading dynamics. Compared with the traditional definition of the betweenness, we adopt two new proposed methods to further evaluate the resilience of several actual networks. We find that some real world networks reach the strongest resilience level against cascading failures in our preferential selection mechanisms of flow destination. Moreover, we explore how to use the minimum cost to maximize the resilience of the studied networks.

  9. Predicting changes in cardiac myocyte contractility during early drug discovery with in vitro assays

    Energy Technology Data Exchange (ETDEWEB)

    Morton, M.J., E-mail: michael.morton@astrazeneca.com [Discovery Sciences, AstraZeneca, Macclesfield, Cheshire SK10 4TG (United Kingdom); Armstrong, D.; Abi Gerges, N. [Drug Safety and Metabolism, AstraZeneca, Macclesfield, Cheshire SK10 4TG (United Kingdom); Bridgland-Taylor, M. [Discovery Sciences, AstraZeneca, Macclesfield, Cheshire SK10 4TG (United Kingdom); Pollard, C.E.; Bowes, J.; Valentin, J.-P. [Drug Safety and Metabolism, AstraZeneca, Macclesfield, Cheshire SK10 4TG (United Kingdom)

    2014-09-01

    Cardiovascular-related adverse drug effects are a major concern for the pharmaceutical industry. Activity of an investigational drug at the L-type calcium channel could manifest in a number of ways, including changes in cardiac contractility. The aim of this study was to define which of the two assay technologies – radioligand-binding or automated electrophysiology – was most predictive of contractility effects in an in vitro myocyte contractility assay. The activity of reference and proprietary compounds at the L-type calcium channel was measured by radioligand-binding assays, conventional patch-clamp, automated electrophysiology, and by measurement of contractility in canine isolated cardiac myocytes. Activity in the radioligand-binding assay at the L-type Ca channel phenylalkylamine binding site was most predictive of an inotropic effect in the canine cardiac myocyte assay. The sensitivity was 73%, specificity 83% and predictivity 78%. The radioligand-binding assay may be run at a single test concentration and potency estimated. The least predictive assay was automated electrophysiology which showed a significant bias when compared with other assay formats. Given the importance of the L-type calcium channel, not just in cardiac function, but also in other organ systems, a screening strategy emerges whereby single concentration ligand-binding can be performed early in the discovery process with sufficient predictivity, throughput and turnaround time to influence chemical design and address a significant safety-related liability, at relatively low cost. - Highlights: • The L-type calcium channel is a significant safety liability during drug discovery. • Radioligand-binding to the L-type calcium channel can be measured in vitro. • The assay can be run at a single test concentration as part of a screening cascade. • This measurement is highly predictive of changes in cardiac myocyte contractility.

  10. Mental Health Screening Center

    Science.gov (United States)

    ... Releases & Announcements Public Service Announcements Partnering with DBSA Mental Health Screening Center These online screening tools are not ... you have any concerns, see your doctor or mental health professional. Depression Screening for Adult Depression Screening for ...

  11. Breast cancer screening

    Science.gov (United States)

    Mammogram - breast cancer screening; Breast exam - breast cancer screening; MRI - breast cancer screening ... is performed to screen women to detect early breast cancer when it is more likely to be cured. ...

  12. Active control of light based on polarization-coupling cascading

    Science.gov (United States)

    Huo, Juan; Zheng, Yuanlin; Chen, Xianfeng

    2014-10-01

    In this letter, we proposed a novel method for optical manipulation based on polarization-coupling cascading in MgO-doped periodically poled lithium niobate crystal. Polarization-coupling cascading, a series of energy exchanges between two orthogonally polarized beams close to phase matching condition, can also lead to phase shifts, in analogy with that in cascaded second-order nonlinearities. In addition, the parameters of light such as phase, amplitude, and group velocity can be modulated by changing the relative power ratio of the incident continuous wave beams. The phase control was demonstrated by Newton's rings experiment, which was in good agreement with the theoretical prediction.

  13. An Improved Phase Disposition SPWM Strategy for Cascaded Multilevel Inverter

    Directory of Open Access Journals (Sweden)

    Jinbang Xu

    2014-01-01

    Full Text Available The Carrier Phase Shifted (CPS strategy is conventional for cascaded multilevel inverter, because it can naturally ensure all cascaded cells to output balanced power. However, in point of spectra of the output line voltage, CPS is suboptimal to Phase Disposition (PD strategy, because the latter can not naturally ensure power balance. This paper presents an improved PD strategy, inspiration from the disposition of CPS strategy triangle carriers. Just reconstructing the triangle carriers of PD strategy, it can not only reserve the waveform quality of the line voltage to be optimal, but also naturally ensure the output power of each cascaded cells to be balanced.

  14. Revised tephra volumes for Cascade Range volcanoes

    Science.gov (United States)

    Nathenson, Manuel

    2017-01-01

    Isopach maps from tephra eruptions from Mount St. Helens were reported in Carey et al. (1995) and for tephra eruptions from Glacier Peak in Gardner et al. (1998). For exponential thinning, the isopach data only define a single slope on a log thickness versus square root of area plot. Carey et al. (1995) proposed a model that was used to estimate a second slope, and volumes were presented in both studies using this model. A study by Sulpizio (2005) for estimating the second slope and square root of area where the lines intersect involves a systematic analysis of many eruptions to provide correlation equations. The purpose of this paper is to recalculate the volumes of Cascades eruptions and compare results from the two methods. In order to gain some perspective on the methods for estimating the second slope, we use data for thickness versus distance beyond the last isopach that are available for some of the larger eruptions in the Cascades. The thickness versus square root of area method is extended to thickness versus distance by developing an approximate relation between the two assuming elliptical isopachs with the source at one of the foci. Based on the comparisons made between the Carey et al. (1995) and Sulpizio (2005) methods, it is felt that the later method provides a better estimate of the second slope. For Mount St. Helens, the estimates of total volume using the Sulpizio (2005) method are generally smaller than those using the Carey et al. (1995) method. For the volume estimates of Carey et al. (1995), the volume of the May 18, 1980, eruption of Mount St. Helens is smaller than six of the eight previous eruptions. With the new volumes using the Sulpizio (2005) method, the 1980 eruption is smaller in volume than the upper end of the range for only three of the layers (Wn, Ye, and Yn) and is the same size as layer We. Thus the 1980 eruption becomes representative of the mid-range of volumes rather than being in the lower range.

  15. Herbivore release through cascading risk effects.

    Science.gov (United States)

    Schmidt-Entling, Martin H; Siegenthaler, Eva

    2009-12-23

    Predators influence prey through consumption, and through trait-mediated effects such as emigration in response to predation risk (risk effects). We studied top-down effects of (sub-) adult wolf spiders (Lycosidae) on arthropods in a meadow. We compared risk effects with the overall top-down effect (including consumption) by gluing the chelicers of wolf spiders to prevent them from killing the prey. In a field experiment, we created three treatments that included either: (i) intact ('predation') wolf spiders; (ii) wolf spiders with glued chelicers ('risk spiders'); or (iii) no (sub-) adult wolf spiders. Young wolf spiders were reduced by their (sub-) adult congeners. Densities of sheetweb spiders (Linyphiidae), a known intraguild prey of wolf spiders, were equally reduced by the presence of risk and predation wolf spiders. Plant- and leafhoppers (Auchenorrhyncha) showed the inverse pattern of higher densities in the presence of both risk and predation wolf spiders. We conclude that (sub-) adult wolf spiders acted as top predators, which reduced densities of intermediate predators and thereby enhanced herbivores. Complementary to earlier studies that found trait-mediated herbivore suppression, our results demonstrate that herbivores can be enhanced through cascading risk effects by top predators.

  16. Particle energy cascade in the intergalactic medium

    Science.gov (United States)

    Valdés, M.; Evoli, C.; Ferrara, A.

    2010-05-01

    We study the development of high-energy (Ein new code MEDEA (Monte Carlo Energy Deposition Analysis) which includes Bremsstrahlung and inverse Compton (IC) processes, along with H/He collisional ionizations and excitations, and electron-electron collisions. The cascade energy partition into heating, excitations and ionizations depends primarily not only on the IGM ionized fraction, xe, but also on redshift, z, due to IC on cosmic microwave background (CMB) photons. While Bremsstrahlung is unimportant under most conditions, IC becomes largely dominant at energies Ein >= 1 MeV. The main effect of IC at injection energies Ein = 1 GeV CMB photons are preferentially upscattered within the X-ray spectrum (hν > 104 eV) and can free stream to the observer. Complete tables of the fractional energy depositions as a function of redshift, Ein and ionized fraction are given. Our results can be used in many astrophysical contexts, with an obvious application related to the study of decaying/annihilating dark matter (DM) candidates in the high-z Universe.

  17. Cascaded Amplitude Modulations in Sound Texture Perception

    Directory of Open Access Journals (Sweden)

    Richard McWalter

    2017-09-01

    Full Text Available Sound textures, such as crackling fire or chirping crickets, represent a broad class of sounds defined by their homogeneous temporal structure. It has been suggested that the perception of texture is mediated by time-averaged summary statistics measured from early auditory representations. In this study, we investigated the perception of sound textures that contain rhythmic structure, specifically second-order amplitude modulations that arise from the interaction of different modulation rates, previously described as “beating” in the envelope-frequency domain. We developed an auditory texture model that utilizes a cascade of modulation filterbanks that capture the structure of simple rhythmic patterns. The model was examined in a series of psychophysical listening experiments using synthetic sound textures—stimuli generated using time-averaged statistics measured from real-world textures. In a texture identification task, our results indicated that second-order amplitude modulation sensitivity enhanced recognition. Next, we examined the contribution of the second-order modulation analysis in a preference task, where the proposed auditory texture model was preferred over a range of model deviants that lacked second-order modulation rate sensitivity. Lastly, the discriminability of textures that included second-order amplitude modulations appeared to be perceived using a time-averaging process. Overall, our results demonstrate that the inclusion of second-order modulation analysis generates improvements in the perceived quality of synthetic textures compared to the first-order modulation analysis considered in previous approaches.

  18. MCDF calculations of Auger cascade processes

    Science.gov (United States)

    Beerwerth, Randolf; Fritzsche, Stephan

    2017-10-01

    We model the multiple ionization of near-neutral core-excited atoms where a cascade of Auger processes leads to the emission of several electrons. We utilize the multiconfiguration Dirac-Fock (MCDF) method to generate approximate wave functions for all fine-structure levels and to account for all decays between them. This approach allows to compute electron spectra, the population of final-states and ion yields, that are accessible in many experiments. Furthermore, our approach is based on the configuration interaction method. A careful treatment of correlation between electronic configurations enables one to model three-electron processes such as an Auger decay that is accompanied by an additional shake-up transition. Here, this model is applied to the triple ionization of atomic cadmium, where we show that the decay of inner-shell 4p holes to triply-charged final states is purely due to the shake-up transition of valence 5s electrons. Contribution to the Topical Issue "Atomic and Molecular Data and their Applications", edited by Gordon W.F. Drake, Jung-Sik Yoon, Daiji Kato, Grzegorz Karwasz.

  19. Quantum cascade lasers (QCLs) in biomedical spectroscopy.

    Science.gov (United States)

    Schwaighofer, Andreas; Brandstetter, Markus; Lendl, Bernhard

    2017-10-02

    Quantum cascade lasers (QCL) are the first room temperature semiconductor laser source for the mid-IR spectral region, triggering substantial development for the advancement of mid-IR spectroscopy. Mid-IR spectroscopy in general provides rapid, label-free and objective analysis, particularly important in the field of biomedical analysis. Due to their unique properties, QCLs offer new possibilities for development of analytical methods to enable quantification of clinically relevant concentration levels and to support medical diagnostics. Compared to FTIR spectroscopy, novel and elaborated measurement techniques can be implemented that allow miniaturized and portable instrumentation. This review illustrates the characteristics of QCLs with a particular focus on their benefits for biomedical analysis. Recent applications of QCL-based spectroscopy for analysis of a variety of clinically relevant samples including breath, urine, blood, interstitial fluid, and biopsy samples are summarized. Further potential for technical advancements is discussed in combination with future prospects for employment of QCL-based devices in routine and point-of-care diagnostics.

  20. Quantum cascade lasers for defense and security

    Science.gov (United States)

    Day, Timothy; Pushkarsky, Michael; Caffey, Dave; Cecchetti, Kristen; Arp, Ron; Whitmore, Alex; Henson, Michael; Takeuchi, Eric B.

    2013-10-01

    Quantum cascade laser (QCL) systems are mature and at the vanguard of a new generation of products that support military applications such as Infrared Countermeasures (IRCM) and targeting. The demanding product requirements for aircraft platforms that include reduced size, weight, power consumption and cost (SWaP-C) extends to portable, battery powered handheld products. QCL technology operates throughout the mid-wave (MWIR) and long-wave (LWIR) infrared to provide new capabilities that leverage existing thermal imaging cameras. In addition to their suitability for aircraft platforms, QCL products are a natural fit to meet operator demands for small, lightweight pointer and beacon capabilities. Field-testing of high power, lightweight, battery operated devices has demonstrated their utility across a range of air and ground applications. This talk will present an overview of QCL technology and the Defense and Security products and capabilities that are enabled by it. This talk will also provide an overview of the extensive environmental and performance testing associated with products based on QCL technology.

  1. Cascading reminiscence bumps in popular music.

    Science.gov (United States)

    Krumhansl, Carol Lynne; Zupnick, Justin Adam

    2013-10-01

    Autobiographical memories are disproportionately recalled for events in late adolescence and early adulthood, a phenomenon called the reminiscence bump. Previous studies on music have found autobiographical memories and life-long preferences for music from this period. In the present study, we probed young adults' personal memories associated with top hits over 5-and-a-half decades, as well as the context of their memories and their recognition of, preference for, quality judgments of, and emotional reactions to that music. All these measures showed the typical increase for music released during the two decades of their lives. Unexpectedly, we found that the same measures peaked for the music of participants' parents' generation. This finding points to the impact of music in childhood and suggests that these results reflect the prevalence of music in the home environment. An earlier peak occurred for 1960s music, which may be explained by its quality or by its transmission through two generations. We refer to this pattern of musical cultural transmission over generations as cascading reminiscence bumps.

  2. Enhanced performance of quantum cascade Raman laser

    Science.gov (United States)

    Yousefvand, Hossein Reza; Aahmadi, Vahid

    2015-05-01

    We present a self-consistent numerical approach for quantum cascade Raman laser (QC-RL) with a modified design to improve the device performance. Our modeling approach is based on monolithic integration of stimulated Raman scattering (SRS) and electrically pumped QC laser. The laser band structure utilizing techniques with both material-dependent effective mass and band nonparabolicity is calculated by solving the Schrodinger-Poisson equations self-consistently. A detailed analysis of output characteristics of the obtained structure is carried out within a simplified 4-level rate equations model taking into account the SRS process. The model accurately explains the operating characteristics found in QCLs, such as damping transient response and non-resonant behavior of modulation frequency response. Furthermore, modification of the structure is focused on improving the SRS in the QC-RL. This leads to an enhancement of the device performance such as threshold current, external quantum efficiency, conversion efficiency, turn-on delay and modulation response. The excellent agreement of the experimental data with the simulated light output-current characteristics confirms the validity of the model.

  3. Optimization of quantum cascade laser operation by geometric design of cascade active band in open and closed models

    Directory of Open Access Journals (Sweden)

    M.V. Tkach

    2013-01-01

    Full Text Available Using the effective mass and rectangular potential approximations, the theory of electron dynamic conductivity is developed for the plane multilayer resonance tunnel structure placed into a constant electric field within the model of open nanosystem, and oscillator forces of quantum transitions within the model of closed nanosystem. For the experimentally produced quantum cascade laser with four-barrier active band of separate cascade, it is proven that just the theory of dynamic conductivity in the model of open cascade most adequately describes the radiation of high frequency electromagnetic field while the electrons transport through the resonance tunnel structure driven by a constant electric field.

  4. Terahertz quantum-cascade lasers based on an interlaced photon-phonon cascade

    Science.gov (United States)

    Köhler, Rüdeger; Tredicucci, Alessandro; Mauro, Cosimo; Beltram, Fabio; Beere, Harvey E.; Linfield, Edmund H.; Davies, A. Giles; Ritchie, David A.

    2004-02-01

    A THz (λ˜80 μm) quantum-cascade laser utilizing alternating photon- and phonon-emitting stages has been developed to achieve efficient extraction of electrons from the lower laser level. Thermal backfilling of electrons is drastically reduced leading to an operation up to 95 K and a weak temperature dependence of the power versus current slope efficiency. The threshold current density is 280 A cm-2 at 6 K and increases to 580 A cm-2 at 90 K. Peak output powers of 10 mW at 30 K and 4 mW at 80 K are obtained.

  5. On periodic orbits in discrete-time cascade systems

    Directory of Open Access Journals (Sweden)

    Huimin Li

    2006-01-01

    Full Text Available We present some results on existence, minimum period, number of periodic orbits, and stability of periodic orbits in discrete-time cascade systems. Some examples are presented to illustrate these results.

  6. Quantum cascade laser infrared spectroscopy of single cancer cells

    KAUST Repository

    Patel, Imran

    2017-03-27

    Quantum cascade laser infrared spectroscopy is a next generation novel imaging technique allowing high resolution spectral imaging of cells. We show after spectral pre-processing, identification of different cancer cell populations within minutes.

  7. Critical Boundary of Cascaded Quadratic Soliton Compression in PPLN

    OpenAIRE

    Guo, Hairun; Zeng, Xianglong; Zhou, Binbin; Bache, Morten

    2012-01-01

    Cascaded quadratic soliton compression in PPLN is investigated and a general critical soliton number is found as the compression boundary. An optimal-parameter diagram for compression at 1550 nm is presented.

  8. Terahertz Quantum Cascade Laser Based 3D Imaging Project

    Data.gov (United States)

    National Aeronautics and Space Administration — LongWave Photonics proposes a terahertz quantum-cascade laser based swept-source optical coherence tomography (THz SS-OCT) system for single-sided, 3D,...

  9. Multicompartment Artificial Organelles Conducting Enzymatic Cascade Reactions inside Cells

    DEFF Research Database (Denmark)

    Gallardo, Maria Godoy; Labay, Cédric Pierre; Trikalitis, Vasileios

    2017-01-01

    capsosome system, which consists of multiple liposomes and fluorescent gold nanoclusters embedded within a polymer carrier capsule. We subsequently demonstrate that encapsulated enzymes preserve their activity intracellularly, allowing for controlled enzymatic cascade reaction within a host cell....

  10. Cascaded spintronic logic with low-dimensional carbon

    Science.gov (United States)

    Friedman, Joseph S.; Girdhar, Anuj; Gelfand, Ryan M.; Memik, Gokhan; Mohseni, Hooman; Taflove, Allen; Wessels, Bruce W.; Leburton, Jean-Pierre; Sahakian, Alan V.

    2017-06-01

    Remarkable breakthroughs have established the functionality of graphene and carbon nanotube transistors as replacements to silicon in conventional computing structures, and numerous spintronic logic gates have been presented. However, an efficient cascaded logic structure that exploits electron spin has not yet been demonstrated. In this work, we introduce and analyse a cascaded spintronic computing system composed solely of low-dimensional carbon materials. We propose a spintronic switch based on the recent discovery of negative magnetoresistance in graphene nanoribbons, and demonstrate its feasibility through tight-binding calculations of the band structure. Covalently connected carbon nanotubes create magnetic fields through graphene nanoribbons, cascading logic gates through incoherent spintronic switching. The exceptional material properties of carbon materials permit Terahertz operation and two orders of magnitude decrease in power-delay product compared to cutting-edge microprocessors. We hope to inspire the fabrication of these cascaded logic circuits to stimulate a transformative generation of energy-efficient computing.

  11. Mutually independent cascades in anisotropic soap-film turbulence

    Science.gov (United States)

    Liu, Chien-Chia; Gioia, Gustavo; Chakraborty, Pinaki

    2015-03-01

    Computational, experimental and field data amassed to date indicate that in 2D turbulence the spectrum of longitudinal velocity fluctuations, E11 (k1) , and the spectrum of transverse velocity fluctuations, E22 (k1) , correspond always to the same cascade, consistent with isotropy, so that E11 (k1) ~k-α and E22 (k1) ~k-α , where the ``spectral exponent'' α is either 5/3 (for the inverse-energy cascade) or 3 (for the enstrophy cascade). Here, we carry out experiments on turbulent 2D soap-film flows in which E11 (k1) ~k - 5 / 3 and E22 (k1) ~k-3 , as if two mutually independent cascades were concurrently active within the same flow. To our knowledge, this species of spectrum has never been observed or predicted theoretically. Our finding might open up new vistas in the understanding of turbulence.

  12. Cascade Structure of Digital Predistorter for Power Amplifier Linearization

    Directory of Open Access Journals (Sweden)

    E. B. Solovyeva

    2015-12-01

    Full Text Available In this paper, a cascade structure of nonlinear digital predistorter (DPD synthesized by the direct learning adaptive algorithm is represented. DPD is used for linearization of power amplifier (PA characteristic, namely for compensation of PA nonlinear distortion. Blocks of the cascade DPD are described by different models: the functional link artificial neural network (FLANN, the polynomial perceptron network (PPN and the radially pruned Volterra model (RPVM. At synthesis of the cascade DPD there is possibility to overcome the ill conditionality problem due to reducing the dimension of DPD nonlinear operator approximation. Results of compensating nonlinear distortion in Wiener–Hammerstein model of PA at the GSM–signal with four carriers are shown. The highest accuracy of PA linearization is produced by the cascade DPD containing PPN and RPVM.

  13. PULSE MODULATION POWER AMPLIFIER WITH ENHANCED CASCADE CONTROL METHOD

    DEFF Research Database (Denmark)

    1998-01-01

    A digital switching power amplifier with Multivariable Enhanced Cascade Controlled (MECC) includes a modulator, a switching power stage and a low pass filter. In the first preferred embodiment an enhanced cascade control structure local to the switching power stage is added, characterised by havi...... and feedback path A to determine stable self-oscillating conditions. An implemented 250W example MECC digital power amplifier has proven superior performance in terms of audio performance (0.005 % distortion, 115 dB dynamic range) and efficiency (92 %).......A digital switching power amplifier with Multivariable Enhanced Cascade Controlled (MECC) includes a modulator, a switching power stage and a low pass filter. In the first preferred embodiment an enhanced cascade control structure local to the switching power stage is added, characterised by having...

  14. Fluorescent and Lanthanide Labeling for Ligand Screens, Assays, and Imaging

    Science.gov (United States)

    Josan, Jatinder S.; De Silva, Channa R.; Yoo, Byunghee; Lynch, Ronald M.; Pagel, Mark D.; Vagner, Josef; Hruby, Victor J.

    2012-01-01

    The use of fluorescent (or luminescent) and metal contrast agents in high-throughput screens, in vitro assays, and molecular imaging procedures has rapidly expanded in recent years. Here we describe the development and utility of high-affinity ligands for cancer theranostics and other in vitro screening studies. In this context, we also illustrate the syntheses and use of heteromultivalent ligands as targeted imaging agents. PMID:21318902

  15. Vortex merging and spectral cascade in two-dimensional flows

    DEFF Research Database (Denmark)

    Nielsen, A.H.; He, X.; Juul Rasmussen, J.

    1996-01-01

    The merging of two identical vortices is studied numerically using a spectral code. It is noted that the enstrophy cascade is most active on the distorted vortex boundaries, with a Kolmogorov-like spectrum E(k) approximate to k(-alpha), alpha less than or equal to 4, developed at high wave numbers....... The inverse energy cascade is completed when the vortices merge into one of larger size. (C) 1996 American Institute of Physics....

  16. The importance of sensory integration processes for action cascading

    OpenAIRE

    Krutika Gohil; Ann-Kathrin Stock; Christian Beste

    2015-01-01

    Dual tasking or action cascading is essential in everyday life and often investigated using tasks presenting stimuli in different sensory modalities. Findings obtained with multimodal tasks are often broadly generalized, but until today, it has remained unclear whether multimodal integration affects performance in action cascading or the underlying neurophysiology. To bridge this gap, we asked healthy young adults to complete a stop-change paradigm which presented different stimuli in either ...

  17. Noise properties and cascadability of SOA-EA regenerators

    DEFF Research Database (Denmark)

    Öhman, Filip; Bischoff, Svend; Tromborg, Bjarne

    2002-01-01

    We suggest and analyse a new device containing concatenated pairs of semiconductor optical amplifiers (SOAs) and electroabsorption modulators (EAs). The device has regenerative properties and improves the cascadability of optical fibre links.......We suggest and analyse a new device containing concatenated pairs of semiconductor optical amplifiers (SOAs) and electroabsorption modulators (EAs). The device has regenerative properties and improves the cascadability of optical fibre links....

  18. Climate Change and Baleen Whale Trophic Cascades in Greenland

    Science.gov (United States)

    2009-09-30

    DISTRIBUTION STATEMENT A: Approved for public release; distribution is unlimited. Climate Change and Baleen Whale Trophic Cascades in Greenland...SUBTITLE Climate Change And Baleen Whale Trophic Cascades In Greenland 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S...2009. Polar bears in Northwest Greenland – An interview survey about the catch and the climate . Meddelelser om Grønland. [In Press, refereed] Heide

  19. Critical Boundary of Cascaded Quadratic Soliton Compression in PPLN

    DEFF Research Database (Denmark)

    Guo, Hairun; Zeng, Xianglong; Zhou, Binbin

    2012-01-01

    Cascaded quadratic soliton compression in PPLN is investigated and a general critical soliton number is found as the compression boundary. An optimal-parameter diagram for compression at 1550 nm is presented.......Cascaded quadratic soliton compression in PPLN is investigated and a general critical soliton number is found as the compression boundary. An optimal-parameter diagram for compression at 1550 nm is presented....

  20. Parton-hadron cascade approach at SPS and RHIC

    Energy Technology Data Exchange (ETDEWEB)

    Nara, Yasushi [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment

    1998-07-01

    A parton-hadron cascade model which is the extension of hadronic cascade model incorporating hard partonic scattering based on HIJING is presented to describe the space-time evolution of parton/hadron system produced by ultra-relativistic nuclear collisions. Hadron yield, baryon stopping and transverse momentum distribution are calculated and compared with HIJING and VNI. Baryon density, energy density and temperature for RHIC are calculated within this model. (author)

  1. Advanced Dementia Research in the Nursing Home: The CASCADE Study

    OpenAIRE

    Mitchell, Susan L.; Kiely, Dan K.; Jones, Richard N.; Prigerson, Holly; Volicer, Ladislav; Teno, Joan M.

    2006-01-01

    Despite the growing number of persons with advanced dementia, and the need to improve their end-of-life care, few studies have addressed this important topic. The objectives of this report are to present the methodology established in the CASCADE (Choices, Attitudes, and Strategies for Care of Advanced Dementia at the End-of-Life) study, and to describe how challenges specific to this research were met. The CASCADE study is an ongoing, federally funded, 5-year prospective cohort study of nurs...

  2. Power system cascading risk assessment based on complex network theory

    Science.gov (United States)

    Wang, Zhuoyang; Hill, David J.; Chen, Guo; Dong, Zhao Yang

    2017-09-01

    When a single failure occurs in a vulnerable part of a power system, this may cause a large area cascading event. Therefore, an advanced method that can assess the risks during cascading events is needed. In this paper, an improved complex network model for power system risk assessment is proposed. Risk is defined by consequence and probability of the failures in this model, which are affected by both power factors and network structure. Compared with existing risk assessment models, the proposed one can evaluate the risk of the system comprehensively during a cascading event by combining the topological and electrical information. A new cascading event simulation module is adopted to identify the power grid cascading chain from a system-level view. In addition, simulations are investigated on the IEEE 14 bus system and IEEE 39 bus system respectively to illustrate the performance of the proposed module. The simulation results demonstrate that the proposed method is effective in a power grid risk assessment during cascading event.

  3. Signal transduction in a covalent post-assembly modification cascade

    Science.gov (United States)

    Pilgrim, Ben S.; Roberts, Derrick A.; Lohr, Thorsten G.; Ronson, Tanya K.; Nitschke, Jonathan R.

    2017-12-01

    Natural reaction cascades control the movement of biomolecules between cellular compartments. Inspired by these systems, we report a synthetic reaction cascade employing post-assembly modification reactions to direct the partitioning of supramolecular complexes between phases. The system is composed of a self-assembled tetrazine-edged FeII8L12 cube and a maleimide-functionalized FeII4L6 tetrahedron. Norbornadiene (NBD) functions as the stimulus that triggers the cascade, beginning with the inverse-electron-demand Diels-Alder reaction of NBD with the tetrazine moieties of the cube. This reaction generates cyclopentadiene as a transient by-product, acting as a relay signal that subsequently undergoes a Diels-Alder reaction with the maleimide-functionalized tetrahedron. Cyclooctyne can selectively inhibit the cascade by outcompeting NBD as the initial trigger. Initiating the cascade with 2-octadecyl NBD leads to selective alkylation of the tetrahedron upon cascade completion. The increased lipophilicity of the C18-tagged tetrahedron drives this complex into a non-polar phase, allowing its isolation from the initially inseparable mixture of complexes.

  4. Cascading off Davis Strait as seen from a NEMO simulation

    Science.gov (United States)

    Marson, Juliana M.; Myers, Paul G.; Hu, Xianmin; Petrie, Brian; Azetsu-Scott, Kumiko; Lee, Craig

    2017-04-01

    Cascading occurs when dense waters form and accumulate on the continental shelf, eventually sliding down the slope and reaching intermediate and deep layers of the ocean. It is an important process for ventilating the deep ocean and capturing carbon from the atmosphere. Although observed in several different locations, especially at high latitudes, cascading has never been reported for the western Greenland shelf. We use the results from a 2002-2014 NEMO ocean model simulation to show that cascading could happen sporadically at Davis Strait. Over the time span of the model run, cascading occurred during five winters with each event starting around February and persisting until the end of May. The occurrence of those events seems to be dependent on (1) the balance between sea ice formation and melting in the region; (2) the amount of freshwater coming from the Arctic through Fram Strait; (3) the presence of West Greenland Irminger Water on the shelf. The cascading water primarily flows into deep Baffin Bay, although not at a sufficient rate to be a primary source of Baffin Bay Deep Water. The simulation's temperature and salinity interannual variability in Davis Strait agrees reasonably with observations (CTD profiles and moorings) but overestimates the peak salinity and density and consequently the strength of the cascade process. Nonetheless, the model simulation indicates that this phenomenon has the potential to occur in this location.

  5. Cascading failures in spatially-embedded random networks.

    Science.gov (United States)

    Asztalos, Andrea; Sreenivasan, Sameet; Szymanski, Boleslaw K; Korniss, Gyorgy

    2014-01-01

    Cascading failures constitute an important vulnerability of interconnected systems. Here we focus on the study of such failures on networks in which the connectivity of nodes is constrained by geographical distance. Specifically, we use random geometric graphs as representative examples of such spatial networks, and study the properties of cascading failures on them in the presence of distributed flow. The key finding of this study is that the process of cascading failures is non-self-averaging on spatial networks, and thus, aggregate inferences made from analyzing an ensemble of such networks lead to incorrect conclusions when applied to a single network, no matter how large the network is. We demonstrate that this lack of self-averaging disappears with the introduction of a small fraction of long-range links into the network. We simulate the well studied preemptive node removal strategy for cascade mitigation and show that it is largely ineffective in the case of spatial networks. We introduce an altruistic strategy designed to limit the loss of network nodes in the event of a cascade triggering failure and show that it performs better than the preemptive strategy. Finally, we consider a real-world spatial network viz. a European power transmission network and validate that our findings from the study of random geometric graphs are also borne out by simulations of cascading failures on the empirical network.

  6. Mitigate Cascading Failures on Networks using a Memetic Algorithm.

    Science.gov (United States)

    Tang, Xianglong; Liu, Jing; Hao, Xingxing

    2016-12-09

    Research concerning cascading failures in complex networks has become a hot topic. However, most of the existing studies have focused on modelling the cascading phenomenon on networks and analysing network robustness from a theoretical point of view, which considers only the damage incurred by the failure of one or several nodes. However, such a theoretical approach may not be useful in practical situation. Thus, we first design a much more practical measure to evaluate the robustness of networks against cascading failures, termed Rcf. Then, adopting Rcf as the objective function, we propose a new memetic algorithm (MA) named MA-Rcf to enhance network the robustness against cascading failures. Moreover, we design a new local search operator that considers the characteristics of cascading failures and operates by connecting nodes with a high probability of having similar loads. In experiments, both synthetic scale-free networks and real-world networks are used to test the efficiency and effectiveness of the MA-Rcf. We systematically investigate the effects of parameters on the performance of the MA-Rcf and validate the performance of the newly designed local search operator. The results show that the local search operator is effective, that MA-Rcf can enhance network robustness against cascading failures efficiently, and that it outperforms existing algorithms.

  7. US physician practices for diagnosing familial hypercholesterolemia: data from the CASCADE-FH registry

    Science.gov (United States)

    Ahmad, Zahid S.; Andersen, Rolf L.; Andersen, Lars H.; O'Brien, Emily C.; Kindt, Iris; Shrader, Peter; Vasandani, Chandna; Newman, Connie B.; deGoma, Emil M.; Baum, Seth J.; Hemphill, Linda C.; Hudgins, Lisa C.; Ahmed, Catherine D.; Kullo, Iftikhar J.; Gidding, Samuel S.; Duffy, Danielle; Neal, William; Wilemon, Katherine; Roe, Matthew T.; Rader, Daniel J.; Ballantyne, Christie M.; Linton, MacRae F.; Duell, P. Barton; Shapiro, Michael D.; Moriarty, Patrick M.; Knowles, Joshua W.

    2017-01-01

    Background In the US familial hypercholesterolemia (FH), patients are underidentified, despite an estimated prevalence of 1:200 to 1:500. Criteria to identify FH patients include Simon Broome, Dutch Lipid Clinic Network (DLCN), or Make Early Diagnosis to Prevent Early Deaths (MEDPED). The use of these criteria in US clinical practices remains unclear. Objective To characterize the FH diagnostic criteria applied by US lipid specialists participating in the FH Foundation's CASCADE FH (CAscade SCreening for Awareness and DEtection of Familial Hypercholesterolemia) patient registry. Methods We performed an observational, cross-sectional analysis of diagnostic criteria chosen for each adult patient, both overall and by baseline patient characteristics, at 15 clinical sites that had contributed data to the registry as of September 8, 2015. A sample of 1867 FH adults was analyzed. The median age at FH diagnosis was 50 years, and the median pretreatment low-density lipoprotein cholesterol (LDL-C) value was 238 mg/dL. The main outcome was the diagnostic criteria chosen. Diagnostic criteria were divided into five nonexclusive categories: “clinical diagnosis,” MEDPED, Simon Broome, DLCN, and other. Results Most adults enrolled in CASCADE FH (55.0%) received a “clinical diagnosis.” The most commonly used formal criteria was Simon–Broome only (21%), followed by multiple diagnostic criteria (16%), MEDPED only (7%), DLCN only (1%), and other (0.5%), P < .0001. Of the patients with only a “clinical diagnosis,” 93% would have met criteria for Simon Broome, DLCN, or MEDPED based on the data available in the registry. Conclusions Our findings demonstrate heterogeneity in the application of FH diagnostic criteria in the United States. A nationwide consensus definition may lead to better identification, earlier treatment, and ultimately CHD prevention. PMID:27678440

  8. Human initiated cascading failures in societal infrastructures.

    Directory of Open Access Journals (Sweden)

    Chris Barrett

    Full Text Available In this paper, we conduct a systematic study of human-initiated cascading failures in three critical inter-dependent societal infrastructures due to behavioral adaptations in response to a crisis. We focus on three closely coupled socio-technical networks here: (i cellular and mesh networks, (ii transportation networks and (iii mobile call networks. In crises, changes in individual behaviors lead to altered travel, activity and calling patterns, which influence the transport network and the loads on wireless networks. The interaction between these systems and their co-evolution poses significant technical challenges for representing and reasoning about these systems. In contrast to system dynamics models for studying these interacting infrastructures, we develop interaction-based models in which individuals and infrastructure elements are represented in detail and are placed in a common geographic coordinate system. Using the detailed representation, we study the impact of a chemical plume that has been released in a densely populated urban region. Authorities order evacuation of the affected area, and this leads to individual behavioral adaptation wherein individuals drop their scheduled activities and drive to home or pre-specified evacuation shelters as appropriate. They also revise their calling behavior to communicate and coordinate among family members. These two behavioral adaptations cause flash-congestion in the urban transport network and the wireless network. The problem is exacerbated with a few, already occurring, road closures. We analyze how extended periods of unanticipated road congestion can result in failure of infrastructures, starting with the servicing base stations in the congested area. A sensitivity analysis on the compliance rate of evacuees shows non-intuitive effect on the spatial distribution of people and on the loading of the base stations. For example, an evacuation compliance rate of 70% results in higher number

  9. Cascaded processing in written compound word production

    Directory of Open Access Journals (Sweden)

    Raymond eBertram

    2015-04-01

    Full Text Available In this study we investigated the intricate interplay between central linguistic processing and peripheral motor processes during typewriting. Participants had to typewrite two-constituent (noun-noun Finnish compounds in response to picture presentation while their typing behavior was registered. As dependent measures we used writing onset time to assess what processes were completed before writing and inter-key intervals to assess what processes were going on during writing. It was found that writing onset time was determined by whole word frequency rather than constituent frequencies, indicating that compound words are retrieved as whole orthographic units before writing is initiated. In addition, we found that the length of the first syllable also affects writing onset time, indicating that the first syllable is fully prepared before writing commences. The inter-key interval results showed that linguistic planning is not fully ready before writing, but cascades into the motor execution phase. More specifically, inter-key intervals were largest at syllable and morpheme boundaries, supporting the view that additional linguistic planning takes place at these boundaries. Bigram and trigram frequency also affected inter-key intervals with shorter intervals corresponding to higher frequencies. This can be explained by stronger memory traces for frequently co-occurring letter sequences in the motor memory for typewriting. These frequency effects were even larger in the second than in the first constituent, indicating that low-level motor memory starts to become more important during the course of writing compound words. We discuss our results in the light of current models of morphological processing and written word production.

  10. Longitudinal gradients along a reservoir cascade

    Science.gov (United States)

    Miranda, L.E.; Habrat, M.D.; Miyazono, S.

    2008-01-01

    Reservoirs have traditionally been regarded as spatially independent entities rather than as longitudinal segments of a river system that are connected upstream and downstream to the river and other reservoirs. This view has frustrated advancement in reservoir science by impeding adequate organization of available information and by hindering interchanges with allied disciplines that often consider impounded rivers at the basin scale. We analyzed reservoir morphology, water quality, and fish assemblage data collected in 24 reservoirs of the Tennessee River; we wanted to describe longitudinal changes occurring at the scale of the entire reservoir series (i.e., cascade) and to test the hypothesis that fish communities and environmental factors display predictable gradients like those recognized for unimpounded rivers. We used a data set collected over a 7-year period; over 3 million fish representing 94 species were included in the data set. Characteristics such as reservoir mean depth, relative size of the limnetic zone, water retention time, oxygen stratification, thermal stratification, substrate size, and water level fluctuations increased in upstream reservoirs. Conversely, reservoir area, extent of riverine and littoral zones, access to floodplains and associated wetlands, habitat diversity, and nutrient and sediment inputs increased in downstream reservoirs. Upstream reservoirs included few, largely lacustrine, ubiquitous fish taxa that were characteristic of the lentic upper reaches of the basin. Fish species richness increased in a downstream direction from 12 to 67 species/ reservoir as riverine species became more common. Considering impoundments at a basin scale by viewing them as sections in a river or links in a chain may generate insight that is not always available when the impoundments are viewed as isolated entities. Basin-scale variables are rarely controllable but constrain the expression of processes at smaller scales and can facilitate the

  11. Dioscin inhibits osteoclast differentiation and bone resorption though down-regulating the Akt signaling cascades

    Energy Technology Data Exchange (ETDEWEB)

    Qu, Xinhua; Zhai, Zanjing; Liu, Xuqiang; Li, Haowei [Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedics, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai (China); Ouyang, Zhengxiao [Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedics, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai (China); Department of Orthopaedics, Hunan Provincial Tumor Hospital and Tumor Hospital of Xiangya School of Medicine, Central South University, Changsha (China); Wu, Chuanlong [Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedics, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai (China); Liu, Guangwang [Department of Orthopaedic Surgery, The Central Hospital of Xuzhou, Affiliated Hospital of Medical Collage of Southeast University, Xuzhou (China); Fan, Qiming; Tang, Tingting [Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedics, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai (China); Qin, An, E-mail: dr.qinan@gmail.com [Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedics, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai (China); Dai, Kerong, E-mail: krdai@163.com [Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedics, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai (China)

    2014-01-10

    Highlights: •A natural-derived compound, dioscin, suppresses osteoclast formation and bone resorption. •Dioscin inhibits osteolytic bone loss in vivo. •Dioscin impairs the Akt signaling cascades pathways during osteoclastogenesis. •Dioscin have therapeutic value in treating osteoclast-related diseases. -- Abstract: Bone resorption is the unique function of osteoclasts (OCs) and is critical for both bone homeostasis and pathologic bone diseases including osteoporosis, rheumatoid arthritis and tumor bone metastasis. Thus, searching for natural compounds that may suppress osteoclast formation and/or function is promising for the treatment of osteoclast-related diseases. In this study, we for the first time demonstrated that dioscin suppressed RANKL-mediated osteoclast differentiation and bone resorption in vitro in a dose-dependent manner. The suppressive effect of dioscin is supported by the reduced expression of osteoclast-specific markers. Further molecular analysis revealed that dioscin abrogated AKT phosphorylation, which subsequently impaired RANKL-induced nuclear factor-kappaB (NF-κB) signaling pathway and inhibited NFATc1 transcriptional activity. Moreover, in vivo studies further verified the bone protection activity of dioscin in osteolytic animal model. Together our data demonstrate that dioscin suppressed RANKL-induced osteoclast formation and function through Akt signaling cascades. Therefore, dioscin is a potential natural agent for the treatment of osteoclast-related diseases.

  12. Engineering of an H2O2auto-scavenging in vivo cascade for pinoresinol production.

    Science.gov (United States)

    Lv, Yongkun; Cheng, Xiaozhong; Du, Guocheng; Zhou, Jingwen; Chen, Jian

    2017-09-01

    Pinoresinol is a natural lignan with a high market value that has potential pharmacological and food supplement applications. Pinoresinol is currently isolated from plants, which suffers from low efficiency and yield. To produce pinoresinol from inexpensive and industrially available eugenol, an in vivo enzymatic cascade composed of vanillyl alcohol oxidase and peroxidase was designed, which scavenges H 2 O 2 automatically and eliminates protein purification and cofactor addition. Two peroxidases were screened and identified from Escherichia coli BL21 (DE3), and tested in the enzymatic cascade. To balance the flux, different genetic architectures were constructed by using ePathBrick and fusion gene approaches. Scavenging H 2 O 2 alleviated by-product toxicity and enzyme inhibition, and led to efficient pinoresinol production. Optimization of the reaction conditions achieved a titer of 11.29 g/L pinoresinol. The molar yield and productivity were 52.77% and 1.03 g/(L × h), respectively. The elegant strategy developed herein utilizes the harmful by-product to drive the biosynthetic reaction forward and simultaneously detoxify cells, thereby preventing enzyme inhibition. Biotechnol. Bioeng. 2017;114: 2066-2074. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  13. Deep-cascade: Cascading 3D Deep Neural Networks for Fast Anomaly Detection and Localization in Crowded Scenes.

    Science.gov (United States)

    Sabokrou, Mohammad; Fayyaz, Mohsen; Fathy, Mahmood; Klette, Reinhard

    2017-02-17

    This paper proposes a fast and reliable method for anomaly detection and localization in video data showing crowded scenes. Time-efficient anomaly localization is an ongoing challenge and subject of this paper. We propose a cubicpatch- based method, characterised by a cascade of classifiers, which makes use of an advanced feature-learning approach. Our cascade of classifiers has two main stages. First, a light but deep 3D auto-encoder is used for early identification of "many" normal cubic patches. This deep network operates on small cubic patches as being the first stage, before carefully resizing remaining candidates of interest, and evaluating those at the second stage using a more complex and deeper 3D convolutional neural network (CNN). We divide the deep autoencoder and the CNN into multiple sub-stages which operate as cascaded classifiers. Shallow layers of the cascaded deep networks (designed as Gaussian classifiers, acting as weak single-class classifiers) detect "simple" normal patches such as background patches, and more complex normal patches are detected at deeper layers. It is shown that the proposed novel technique (a cascade of two cascaded classifiers) performs comparable to current top-performing detection and localization methods on standard benchmarks, but outperforms those in general with respect to required computation time.

  14. Chlorinated Glycopeptide Antibiotic Peptide Precursors Improve Cytochrome P450-Catalyzed Cyclization Cascade Efficiency.

    Science.gov (United States)

    Peschke, Madeleine; Brieke, Clara; Goode, Rob J A; Schittenhelm, Ralf B; Cryle, Max J

    2017-03-07

    The activity of glycopeptide antibiotics (GPAs) depends upon important structural modifications to their precursor heptapeptide backbone: specifically, the cytochrome P450-catalyzed oxidative cross-linking of aromatic side chains as well as the halogenation of specific residues within the peptide. The timing of halogenation and its effect on the cyclization of the peptide are currently unclear. Our results show that chlorination of peptide precursors improves their processing by P450 enzymes in vitro, which provides support for GPA halogenation occurring prior to peptide cyclization during nonribosomal peptide synthesis. We could also determine that the activity of the second enzyme in the oxidative cyclization cascade, OxyA, remains higher for chlorinated peptide substrates even when the biosynthetic GPA product possesses an altered chlorination pattern, which supports the role of the chlorine atoms in orienting the peptide substrate in the active site of these enzymes.

  15. Nitric oxide: a downstream mediator of calcium toxicity in the ischemic cascade.

    Science.gov (United States)

    Maiese, K; Wagner, J; Boccone, L

    1994-01-17

    Loss of cellular calcium homeostasis or the production of nitric oxide (NO) have been cited as possible mechanisms that may contribute to neuronal degeneration during ischemia. We therefore examined whether cellular calcium blockade, using the agent HA1077, was protective during anoxia in hippocampal neuronal cell cultures, and whether the in vitro effects of this drug were linked to the NO pathway. Administration of the agent during anoxia was neuroprotective in neuronal cell culture. In contrast, HA1077 did not protect hippocampal neurons during NO exposure. In addition, inhibition of NO synthesis in conjunction with HA1077 application during anoxia did not significantly increase survival beyond the maximum protection afforded by HA1077 alone. These results suggest that calcium may be an initial messenger in the ischemic cascade, but that subsequent neuronal degeneration is dependent upon the NO pathway.

  16. Preliminary phytochemical screening and antibacterial testing of the ...

    African Journals Online (AJOL)

    The ethanolic and petroleum ether extracts of Piliostigma thonningii (Schum) used traditionally for relieving toothache and cough were screened for their phytochemical constituents and in vitro antibacterial activity. Chemical and chromatographic methods were used for the phytochemical screening of the extracts. Inhibitory ...

  17. Screening of the Open Source Malaria Box Reveals an Early Lead Compound for the Treatment of Alveolar Echinococcosis.

    Directory of Open Access Journals (Sweden)

    Britta Stadelmann

    2016-03-01

    Full Text Available The metacestode (larval stage of the tapeworm Echinococcus multilocularis causes alveolar echinococcosis (AE, a very severe and in many cases incurable disease. To date, benzimidazoles such as albendazole and mebendazole are the only approved chemotherapeutical treatment options. Benzimidazoles inhibit metacestode proliferation, but do not act parasiticidal. Thus, benzimidazoles have to be taken a lifelong, can cause adverse side effects such as hepatotoxicity, and are ineffective in some patients. We here describe a newly developed screening cascade for the evaluation of the in vitro efficacy of new compounds that includes assessment of parasiticidal activity. The Malaria Box from Medicines for Malaria Venture (MMV, comprised of 400 commercially available chemicals that show in vitro activity against Plasmodium falciparum, was repurposed. Primary screening was carried out at 10 μM by employing the previously described PGI assay, and resulted in the identification of 24 compounds that caused physical damage in metacestodes. Seven out of these 24 drugs were also active at 1 μM. Dose-response assays revealed that only 2 compounds, namely MMV665807 and MMV665794, exhibited an EC50 value below 5 μM. Assessments using human foreskin fibroblasts and Reuber rat hepatoma cells showed that the salicylanilide MMV665807 was less toxic for these two mammalian cell lines than for metacestodes. The parasiticidal activity of MMV665807 was then confirmed using isolated germinal layer cell cultures as well as metacestode vesicles by employing viability assays, and its effect on metacestodes was morphologically evaluated by electron microscopy. However, both oral and intraperitoneal application of MMV665807 to mice experimentally infected with E. multilocularis metacestodes did not result in any reduction of the parasite load.

  18. Screening of the Open Source Malaria Box Reveals an Early Lead Compound for the Treatment of Alveolar Echinococcosis.

    Science.gov (United States)

    Stadelmann, Britta; Rufener, Reto; Aeschbacher, Denise; Spiliotis, Markus; Gottstein, Bruno; Hemphill, Andrew

    2016-03-01

    The metacestode (larval) stage of the tapeworm Echinococcus multilocularis causes alveolar echinococcosis (AE), a very severe and in many cases incurable disease. To date, benzimidazoles such as albendazole and mebendazole are the only approved chemotherapeutical treatment options. Benzimidazoles inhibit metacestode proliferation, but do not act parasiticidal. Thus, benzimidazoles have to be taken a lifelong, can cause adverse side effects such as hepatotoxicity, and are ineffective in some patients. We here describe a newly developed screening cascade for the evaluation of the in vitro efficacy of new compounds that includes assessment of parasiticidal activity. The Malaria Box from Medicines for Malaria Venture (MMV), comprised of 400 commercially available chemicals that show in vitro activity against Plasmodium falciparum, was repurposed. Primary screening was carried out at 10 μM by employing the previously described PGI assay, and resulted in the identification of 24 compounds that caused physical damage in metacestodes. Seven out of these 24 drugs were also active at 1 μM. Dose-response assays revealed that only 2 compounds, namely MMV665807 and MMV665794, exhibited an EC50 value below 5 μM. Assessments using human foreskin fibroblasts and Reuber rat hepatoma cells showed that the salicylanilide MMV665807 was less toxic for these two mammalian cell lines than for metacestodes. The parasiticidal activity of MMV665807 was then confirmed using isolated germinal layer cell cultures as well as metacestode vesicles by employing viability assays, and its effect on metacestodes was morphologically evaluated by electron microscopy. However, both oral and intraperitoneal application of MMV665807 to mice experimentally infected with E. multilocularis metacestodes did not result in any reduction of the parasite load.

  19. Aeration efficiency over stepped cascades: better predictions from flow regimes.

    Science.gov (United States)

    Khdhiri, Hatem; Potier, Olivier; Leclerc, Jean-Pierre

    2014-05-15

    Stepped cascades are recognized as high potential air-water gas exchangers. In natural rivers, these structures enhance oxygen transfer to water by creating turbulence at interface with increasing air entrainment in water and air-water surface exchange. Stepped cascades could be really useful to improve the natural self-purification process by providing oxygen to aerobic micro-organisms. The aeration performance of these structures depends on several operating and geometrical parameters. In the literature, several empirical correlations for aeration efficiency prediction on stepped cascades exist. Most of these correlations are only applicable for operating and geometrical parameters in the range of which they have been developed. In this paper, 398 experimental sets of data (from our experiments and collected from literature) were used to develop a correlation for aeration prediction over stepped cascades derived from dimensional analysis and parameterized for each individual flow regime in order to consider change in flow regime effect on oxygen transfer. This new correlation allowed calculating the whole set of data obtained for cascades with steps heights between 0.05 m and 0.254 m, cascade total height between 0.25 m and 2.5 m, for discharges per unit of width ranging from 0.28 10(-3) m(2)/s to 600 10(-3) m(2)/s and for cascade steps number between 3 and 25. In these ranges of parameters, standard deviation for aeration efficiency estimation was found to be less than 17%. Finally, advices were proposed to help and improve the structure design in order to improve aeration. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. In vitro

    Science.gov (United States)

    Augustine, Dominic; Rao, Roopa S; Anbu, Jayaraman; Chidambara Murthy, K N

    2017-12-01

    The earthworm coelomic fluid (ECF) has shown proven antiproliferative effect against breast, liver, gastrointestinal, and brain cancer, but it is least explored in oral cancer. The present in vitro study is an attempt to investigate the antiproliferative activity of ECF on oral cancer cell line squamous cell carcinoma (SCC)-9. ECF was collected from the species Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) stored at -80°C. Percentage inhibition of ECF on squamous cell carcinoma-9 cells in vitro was recorded at 24 h. Protein estimation was done using Bradford protein assay validated by the biuret method. Cytotoxicity was tested at 2.5, 5, 10, 20, 40, and 80 μg/ml concentrations by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay in SCC-9 cells in vitro . GraphPad Prism 7.0 software was used to calculate the inhibitory concentration (IC 50 ). Chi-square test was used to analyze the difference between samples. The test samples EE, EF, and PE inhibited the growth of SCC-9 cells significantly in a dose-dependent manner, and the IC 50 values were found to be 4.6, 44.69, and 5.27 μg/ml, respectively. The antiproliferative effect was found to be variable among the three earthworm species with EE showing the most promising effect followed by PE and EF. Establishing the antiproliferative effect of ECF on oral cancer cells could be an initial step toward drug development and future anticancer research. The preliminary investigation has shown that ECF has a promising antiproliferative effect on oral cancer cells in vitro . The present pilot study evaluated the in vitro antiproliferative effect of earthworm coelomic fluid (ECF) of Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) on squamous cell carcinoma-9 cell line. The ECF inhibitory activity was promising at inhibitory concentration values of 4.6, 44.69, and 5.27 μg/ml, respectively. Further studies pertaining to antiproliferative mechanism of EE

  1. Cascaded systems analysis of photon counting detectors.

    Science.gov (United States)

    Xu, J; Zbijewski, W; Gang, G; Stayman, J W; Taguchi, K; Lundqvist, M; Fredenberg, E; Carrino, J A; Siewerdsen, J H

    2014-10-01

    Photon counting detectors (PCDs) are an emerging technology with applications in spectral and low-dose radiographic and tomographic imaging. This paper develops an analytical model of PCD imaging performance, including the system gain, modulation transfer function (MTF), noise-power spectrum (NPS), and detective quantum efficiency (DQE). A cascaded systems analysis model describing the propagation of quanta through the imaging chain was developed. The model was validated in comparison to the physical performance of a silicon-strip PCD implemented on an experimental imaging bench. The signal response, MTF, and NPS were measured and compared to theory as a function of exposure conditions (70 kVp, 1-7 mA), detector threshold, and readout mode (i.e., the option for coincidence detection). The model sheds new light on the dependence of spatial resolution, charge sharing, and additive noise effects on threshold selection and was used to investigate the factors governing PCD performance, including the fundamental advantages and limitations of PCDs in comparison to energy-integrating detectors (EIDs) in the linear regime for which pulse pileup can be ignored. The detector exhibited highly linear mean signal response across the system operating range and agreed well with theoretical prediction, as did the system MTF and NPS. The DQE analyzed as a function of kilovolt (peak), exposure, detector threshold, and readout mode revealed important considerations for system optimization. The model also demonstrated the important implications of false counts from both additive electronic noise and charge sharing and highlighted the system design and operational parameters that most affect detector performance in the presence of such factors: for example, increasing the detector threshold from 0 to 100 (arbitrary units of pulse height threshold roughly equivalent to 0.5 and 6 keV energy threshold, respectively), increased the f50 (spatial-frequency at which the MTF falls to a value of

  2. The HIV care cascade: Japanese perspectives.

    Directory of Open Access Journals (Sweden)

    Aikichi Iwamoto

    Full Text Available Japan has been known as a low HIV-prevalence country with a concentrated epidemic among high-risk groups. However, it has not been determined whether Japan meets the 90-90-90 goals set by the Joint United Nations Programme on HIV/AIDS (UNAIDS/World Health Organization (WHO. Moreover, to date, the HIV care cascade has not been examined. We estimated the total number of diagnosed people living with HIV/AIDS (PLWHA (n = 22,840 based on legal reports to the Ministry of Health, Labour and Welfare by subtracting the number of foreigners who left Japan (n = 2,273 and deaths (n = 2,321 from the cumulative diagnosis report (n = 27,434. The number of total undiagnosed PLWHA was estimated by age and sex specific HIV-positive rates observed among first-time blood donors between 2011-2015 in Japan. Our estimates show that 14.4% (n = 3,830 of all PLWHA (n = 26,670 were undiagnosed in Japan at the end of 2015. The number of patients retained in care (n = 20,615: 77.3% of PLWHA, the percentage of those on antiretroviral therapy (n = 18,921: 70.9% of PLWHA and those with suppressed viral loads (<200 copies/mL; n = 18,756: 70.3% of PLWHA were obtained through a questionnaire survey conducted in the AIDS Core Hospitals throughout the country. According to these estimates, Japan failed to achieve the first two of the three UNAIDS/WHO targets (22,840/26,670 = 85.6% of HIV-positive cases were diagnosed; 18,921/22,840 = 82.8% of those diagnosed were treated; 18,756/18,921 = 99.1% of those treated experienced viral suppression. Although the antiretroviral treatment uptake and success after retention in medical care appears to be excellent in Japan, there are unmet needs, mainly at the surveillance level before patients are retained in care. The promotion of HIV testing and treatment programs among the key affected populations (especially men who have sex with men may contribute to further decreasing the HIV epidemic and achieving the UNAIDS/WHO targets in Japan.

  3. Drop-out from the tuberculosis contact investigation cascade in a routine public health setting in urban Uganda: A prospective, multi-center study.

    Science.gov (United States)

    Armstrong-Hough, Mari; Turimumahoro, Patricia; Meyer, Amanda J; Ochom, Emmanuel; Babirye, Diana; Ayakaka, Irene; Mark, David; Ggita, Joseph; Cattamanchi, Adithya; Dowdy, David; Mugabe, Frank; Fair, Elizabeth; Haberer, Jessica E; Katamba, Achilles; Davis, J Lucian

    2017-01-01

    Seven public tuberculosis (TB) units in Kampala, Uganda, where Uganda's national TB program recently introduced household contact investigation, as recommended by 2012 guidelines from WHO. To apply a cascade analysis to implementation of household contact investigation in a programmatic setting. Prospective, multi-center observational study. We constructed a cascade for household contact investigation to describe the proportions of: 1) index patient households recruited; 2) index patient households visited; 3) contacts screened for TB; and 4) contacts completing evaluation for, and diagnosed with, active TB. 338 (33%) of 1022 consecutive index TB patients were eligible for contact investigation. Lay health workers scheduled home visits for 207 (61%) index patients and completed 104 (50%). Among 287 eligible contacts, they screened 256 (89%) for symptoms or risk factors for TB. 131 (51%) had an indication for further TB evaluation. These included 59 (45%) with symptoms alone, 58 (44%) children contacts found to be symptomatic or at risk, 26 (20%) contacts completed evaluation, including five (19%) diagnosed with and treated for active TB, for an overall yield of 1.7%. The cumulative conditional probability of completing the entire cascade was 5%. Major opportunities exist for improving the effectiveness and yield of TB contact investigation by increasing the proportion of index households completing screening visits by lay health workers and the proportion of at-risk contacts completing TB evaluation.

  4. Active Cascade Screening in Primary Inherited Arrhythmia Syndromes : Does It Lead to Prophylactic Treatment?

    NARCIS (Netherlands)

    Hofman, Nynke; Tan, Hanno L.; Alders, Marielle; van Langen, Irene M.; Wilde, Arthur A.M.

    2010-01-01

    Objectives: The purpose of this study was to investigate the follow-up and treatment of the mutation-carrying relatives of a proband with an inherited arrhythmia syndrome. Background: The congenital long QT syndrome (LQTS), catecholaminergic polymorphic ventricular tachycardia (CPVT), and Brugada

  5. RBC Antibody Screen

    Science.gov (United States)

    ... products and services. Advertising & Sponsorship: Policy | Opportunities RBC Antibody Screen Share this page: Was this page helpful? ... Indirect Coombs Test; Indirect Anti-human Globulin Test; Antibody Screen Formal name: Red Blood Cell Antibody Screen ...

  6. Screen time and children

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/patientinstructions/000355.htm Screen time and children To use the sharing features on ... videos is considered unhealthy screen time. Current Screen Time Guidelines Children under age 2 should have no ...

  7. Prenatal Genetic Screening Tests

    Science.gov (United States)

    ... Education & Events Advocacy For Patients About ACOG Prenatal Genetic Screening Tests Home For Patients Search FAQs Prenatal ... Screening Tests FAQ165, July 2017 PDF Format Prenatal Genetic Screening Tests Pregnancy What is prenatal genetic testing? ...

  8. Endometrial Cancer Screening

    Science.gov (United States)

    ... Treatment Endometrial Cancer Prevention Endometrial Cancer Screening Research Endometrial Cancer Screening (PDQ®)–Patient Version What is screening? ... These are called diagnostic tests . General Information About Endometrial Cancer Key Points Endometrial cancer is a disease ...

  9. Stomach (Gastric) Cancer Screening

    Science.gov (United States)

    ... Stomach Cancer Prevention Stomach Cancer Screening Research Stomach (Gastric) Cancer Screening (PDQ®)–Patient Version What is screening? Go ... are called diagnostic tests . General Information About Stomach (Gastric) Cancer Key Points Stomach cancer is a disease in ...

  10. Cascades and Dissipative Anomalies in Compressible Fluid Turbulence

    Science.gov (United States)

    Eyink, Gregory L.; Drivas, Theodore D.

    2018-02-01

    We investigate dissipative anomalies in a turbulent fluid governed by the compressible Navier-Stokes equation. We follow an exact approach pioneered by Onsager, which we explain as a nonperturbative application of the principle of renormalization-group invariance. In the limit of high Reynolds and Péclet numbers, the flow realizations are found to be described as distributional or "coarse-grained" solutions of the compressible Euler equations, with standard conservation laws broken by turbulent anomalies. The anomalous dissipation of kinetic energy is shown to be due not only to local cascade but also to a distinct mechanism called pressure-work defect. Irreversible heating in stationary, planar shocks with an ideal-gas equation of state exemplifies the second mechanism. Entropy conservation anomalies are also found to occur via two mechanisms: an anomalous input of negative entropy (negentropy) by pressure work and a cascade of negentropy to small scales. We derive "4 /5 th-law"-type expressions for the anomalies, which allow us to characterize the singularities (structure-function scaling exponents) required to sustain the cascades. We compare our approach with alternative theories and empirical evidence. It is argued that the "Big Power Law in the Sky" observed in electron density scintillations in the interstellar medium is a manifestation of a forward negentropy cascade or an inverse cascade of usual thermodynamic entropy.

  11. Trend-driven information cascades on random networks.

    Science.gov (United States)

    Kobayashi, Teruyoshi

    2015-12-01

    Threshold models of global cascades have been extensively used to model real-world collective behavior, such as the contagious spread of fads and the adoption of new technologies. A common property of those cascade models is that a vanishingly small seed fraction can spread to a finite fraction of an infinitely large network through local infections. In social and economic networks, however, individuals' behavior is often influenced not only by what their direct neighbors are doing, but also by what the majority of people are doing as a trend. A trend affects individuals' behavior while individuals' behavior creates a trend. To analyze such a complex interplay between local- and global-scale phenomena, I generalize the standard threshold model by introducing a type of node called global nodes (or trend followers), whose activation probability depends on a global-scale trend, specifically the percentage of activated nodes in the population. The model shows that global nodes play a role as accelerating cascades once a trend emerges while reducing the probability of a trend emerging. Global nodes thus either facilitate or inhibit cascades, suggesting that a moderate share of trend followers may maximize the average size of cascades.

  12. Gradients in Catostomid assemblages along a reservoir cascade

    Science.gov (United States)

    Miranda, Leandro E.; Keretz, Kevin R.; Gilliland, Chelsea R.

    2017-01-01

    Serial impoundment of major rivers leads to alterations of natural flow dynamics and disrupts longitudinal connectivity. Catostomid fishes (suckers, family Catostomidae) are typically found in riverine or backwater habitats yet are able to persist in impounded river systems. To the detriment of conservation, there is limited information about distribution of catostomid fishes in impounded rivers. We examined the longitudinal distribution of catostomid fishes over 23 reservoirs of the Tennessee River reservoir cascade, encompassing approximately 1600 km. Our goal was to develop a basin-scale perspective to guide conservation efforts. Catostomid species composition and assemblage structure changed longitudinally along the reservoir cascade. Catostomid species biodiversity was greatest in reservoirs lower in the cascade. Assemblage composition shifted from dominance by spotted sucker Minytrema melanops and buffalos Ictiobus spp. in the lower reservoirs to carpsuckers Carpiodes spp. midway through the cascade and redhorses Moxostoma spp. in the upper reservoirs. Most species did not extend the length of the cascade, and some species were rare, found in low numbers and in few reservoirs. The observed gradients in catostomid assemblages suggest the need for basin-scale conservation measures focusing on three broad areas: (1) conservation and management of the up-lake riverine reaches of the lower reservoirs, (2) maintenance of the access to quality habitat in tributaries to the upper reservoirs and (3) reintroductions into currently unoccupied habitat within species' historic distributions

  13. INCAS: an analytical model to describe displacement cascades

    Science.gov (United States)

    Jumel, Stéphanie; Claude Van-Duysen, Jean

    2004-07-01

    REVE (REactor for Virtual Experiments) is an international project aimed at developing tools to simulate neutron irradiation effects in Light Water Reactor materials (Fe, Ni or Zr-based alloys). One of the important steps of the project is to characterise the displacement cascades induced by neutrons. Accordingly, the Department of Material Studies of Electricité de France developed an analytical model based on the binary collision approximation. This model, called INCAS (INtegration of CAScades), was devised to be applied on pure elements; however, it can also be used on diluted alloys (reactor pressure vessel steels, etc.) or alloys composed of atoms with close atomic numbers (stainless steels, etc.). INCAS describes displacement cascades by taking into account the nuclear collisions and electronic interactions undergone by the moving atoms. In particular, it enables to determine the mean number of sub-cascades induced by a PKA (depending on its energy) as well as the mean energy dissipated in each of them. The experimental validation of INCAS requires a large effort and could not be carried out in the framework of the study. However, it was verified that INCAS results are in conformity with those obtained from other approaches. As a first application, INCAS was applied to determine the sub-cascade spectrum induced in iron by the neutron spectrum corresponding to the central channel of the High Flux Irradiation Reactor of Oak Ridge National Laboratory.

  14. INCAS: an analytical model to describe displacement cascades

    Energy Technology Data Exchange (ETDEWEB)

    Jumel, Stephanie E-mail: stephanie.jumel@edf.fr; Claude Van-Duysen, Jean E-mail: jean-claude.van-duysen@edf.fr

    2004-07-01

    REVE (REactor for Virtual Experiments) is an international project aimed at developing tools to simulate neutron irradiation effects in Light Water Reactor materials (Fe, Ni or Zr-based alloys). One of the important steps of the project is to characterise the displacement cascades induced by neutrons. Accordingly, the Department of Material Studies of Electricite de France developed an analytical model based on the binary collision approximation. This model, called INCAS (INtegration of CAScades), was devised to be applied on pure elements; however, it can also be used on diluted alloys (reactor pressure vessel steels, etc.) or alloys composed of atoms with close atomic numbers (stainless steels, etc.). INCAS describes displacement cascades by taking into account the nuclear collisions and electronic interactions undergone by the moving atoms. In particular, it enables to determine the mean number of sub-cascades induced by a PKA (depending on its energy) as well as the mean energy dissipated in each of them. The experimental validation of INCAS requires a large effort and could not be carried out in the framework of the study. However, it was verified that INCAS results are in conformity with those obtained from other approaches. As a first application, INCAS was applied to determine the sub-cascade spectrum induced in iron by the neutron spectrum corresponding to the central channel of the High Flux Irradiation Reactor of Oak Ridge National Laboratory.

  15. Reducing Cascading Failure Risk by Increasing Infrastructure Network Interdependence

    Science.gov (United States)

    Korkali, Mert; Veneman, Jason G.; Tivnan, Brian F.; Bagrow, James P.; Hines, Paul D. H.

    2017-01-01

    Increased interconnection between critical infrastructure networks, such as electric power and communications systems, has important implications for infrastructure reliability and security. Others have shown that increased coupling between networks that are vulnerable to internetwork cascading failures can increase vulnerability. However, the mechanisms of cascading in these models differ from those in real systems and such models disregard new functions enabled by coupling, such as intelligent control during a cascade. This paper compares the robustness of simple topological network models to models that more accurately reflect the dynamics of cascading in a particular case of coupled infrastructures. First, we compare a topological contagion model to a power grid model. Second, we compare a percolation model of internetwork cascading to three models of interdependent power-communication systems. In both comparisons, the more detailed models suggest substantially different conclusions, relative to the simpler topological models. In all but the most extreme case, our model of a “smart” power network coupled to a communication system suggests that increased power-communication coupling decreases vulnerability, in contrast to the percolation model. Together, these results suggest that robustness can be enhanced by interconnecting networks with complementary capabilities if modes of internetwork failure propagation are constrained. PMID:28317835

  16. No beneficial effect of preimplantation genetic screening in women of advanced maternal age with a high risk for embryonic aneuploidy

    NARCIS (Netherlands)

    Twisk, Moniek; Mastenbroek, Sebastiaan; Hoek, Annemieke; Heineman, Maas-Jan; van der Veen, Fulco; Bossuyt, Patrick M.; Repping, Sjoerd; Korevaar, Johanna C.

    2008-01-01

    Human preimplantation embryos generated through in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) treatments show a variable rate of numerical chromosome abnormalities or aneuploidies. Preimplantation genetic screening (PGS) has been designed to screen for aneuploidies in high

  17. Substrate specificity analysis of protein kinase complex Dbf2-Mob1 by peptide library and proteome array screening

    Directory of Open Access Journals (Sweden)

    Snyder Michael

    2005-10-01

    Full Text Available Abstract Background The mitotic exit network (MEN is a group of proteins that form a signaling cascade that is essential for cells to exit mitosis in Saccharomyces cerevisiae. The MEN has also been implicated in playing a role in cytokinesis. Two components of this signaling pathway are the protein kinase Dbf2 and its binding partner essential for its kinase activity, Mob1. The components of MEN that act upstream of Dbf2-Mob1 have been characterized, but physiological substrates for Dbf2-Mob1 have yet to be identified. Results Using a combination of peptide library selection, phosphorylation of opitmal peptide variants, and screening of a phosphosite array, we found that Dbf2-Mob1 preferentially phosphorylated serine over threonine and required an arginine three residues upstream of the phosphorylated serine in its substrate. This requirement for arginine in peptide substrates could not be substituted with the similarly charged lysine. This specificity determined for peptide substrates was also evident in many of the proteins phosphorylated by Dbf2-Mob1 in a proteome chip analysis. Conclusion We have determined by peptide library selection and phosphosite array screening that the protein kinase Dbf2-Mob1 preferentially phosphorylated substrates that contain an RXXS motif. A subsequent proteome microarray screen revealed proteins that can be phosphorylated by Dbf2-Mob1 in vitro. These proteins are enriched for RXXS motifs, and may include substrates that mediate the function of Dbf2-Mob1 in mitotic exit and cytokinesis. The relatively low degree of sequence restriction at the site of phosphorylation suggests that Dbf2 achieves specificity by docking its substrates at a site that is distinct from the phosphorylation site

  18. Cascaded VLSI Chips Help Neural Network To Learn

    Science.gov (United States)

    Duong, Tuan A.; Daud, Taher; Thakoor, Anilkumar P.

    1993-01-01

    Cascading provides 12-bit resolution needed for learning. Using conventional silicon chip fabrication technology of VLSI, fully connected architecture consisting of 32 wide-range, variable gain, sigmoidal neurons along one diagonal and 7-bit resolution, electrically programmable, synaptic 32 x 31 weight matrix implemented on neuron-synapse chip. To increase weight nominally from 7 to 13 bits, synapses on chip individually cascaded with respective synapses on another 32 x 32 matrix chip with 7-bit resolution synapses only (without neurons). Cascade correlation algorithm varies number of layers effectively connected into network; adds hidden layers one at a time during learning process in such way as to optimize overall number of neurons and complexity and configuration of network.

  19. A Cascade-Based Emergency Model for Water Distribution Network

    Directory of Open Access Journals (Sweden)

    Qing Shuang

    2015-01-01

    Full Text Available Water distribution network is important in the critical physical infrastructure systems. The paper studies the emergency resource strategies on water distribution network with the approach of complex network and cascading failures. The model of cascade-based emergency for water distribution network is built. The cascade-based model considers the network topology analysis and hydraulic analysis to provide a more realistic result. A load redistribution function with emergency recovery mechanisms is established. From the aspects of uniform distribution, node betweenness, and node pressure, six recovery strategies are given to reflect the network topology and the failure information, respectively. The recovery strategies are evaluated with the complex network indicators to describe the failure scale and failure velocity. The proposed method is applied by an illustrative example. The results showed that the recovery strategy considering the node pressure can enhance the network robustness effectively. Besides, this strategy can reduce the failure nodes and generate the least failure nodes per time.

  20. Bearing-Only Formation Control for Cascade Multirobots

    Directory of Open Access Journals (Sweden)

    Qing Han

    2016-01-01

    Full Text Available A new formation control method is proposed, which is used to queue multirobots in a single-direction cascade structure. In the cascade formation, each robot is a follower for the previous robot and a leader for the next robot, and the robots in the middle act as both leader and follower. The follower robot can only observe the bearing information of the leader robot. The observability of the cascade leader-follower formation is studied, which shows that the bearing-only observation meets the observability conditions required for the nonlinear system. Based on the bearing-only observations, the unscented Kalman filter (UKF is employed for the state estimation of the leader and the follower robots at all levels, which enables the real-time movement control of the follower robots via the input-output feedback control. Simulation results demonstrate that the proposed approach can efficiently control the formation of multirobots as desired.