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Sample records for viable cells recovered

  1. A viable foal obtained by equine somatic cell nuclear transfer using oocytes recovered from immature follicles of live mares.

    Science.gov (United States)

    Choi, Young-Ho; Norris, Jody D; Velez, Isabel C; Jacobson, Candace C; Hartman, David L; Hinrichs, Katrin

    2013-03-15

    The presence of heterogenous mitochondria from the host ooplast affects the acceptance of offspring obtained by somatic cell nuclear transfer. This might be avoided by obtaining oocytes from selected females, but is then complicated by low numbers of available oocytes. We examined the efficiency of equine somatic cell nuclear transfer using oocytes recovered by transvaginal aspiration of immature follicles from 11 mares. Use of metaphase I oocytes as cytoplasts and of scriptaid (a histone deacetylase inhibitor) treatment during oocyte activation were evaluated to determine if these approaches would increase blastocyst production. In experiment 1, blastocyst development was 0/14 for metaphase I oocytes and 4/103 (4%) for metaphase II oocytes. Three blastocysts were transferred to recipient mares, resulting in two pregnancies and one live foal, which died shortly after birth. In experiment 2, blastocyst development was 2/47 (4%) for control oocytes and 1/83 (1%) for scriptaid-treated oocytes. No foals were born from two blastocysts transferred in the control group. The blastocyst from the scriptaid treatment resulted in birth of a live foal. In conclusion, this is apparently the first report of production of a viable cloned foal from oocytes collected from immature follicles of live mares, supporting the possibility of cloning using oocytes from selected mares. Copyright © 2013 Elsevier Inc. All rights reserved.

  2. Viable Cell Culture Banking for Biodiversity Characterization and Conservation.

    Science.gov (United States)

    Ryder, Oliver A; Onuma, Manabu

    2018-02-15

    Because living cells can be saved for indefinite periods, unprecedented opportunities for characterizing, cataloging, and conserving biological diversity have emerged as advanced cellular and genetic technologies portend new options for preventing species extinction. Crucial to realizing the potential impacts of stem cells and assisted reproductive technologies on biodiversity conservation is the cryobanking of viable cell cultures from diverse species, especially those identified as vulnerable to extinction in the near future. The advent of in vitro cell culture and cryobanking is reviewed here in the context of biodiversity collections of viable cell cultures that represent the progress and limitations of current efforts. The prospects for incorporating collections of frozen viable cell cultures into efforts to characterize the genetic changes that have produced the diversity of species on Earth and contribute to new initiatives in conservation argue strongly for a global network of facilities for establishing and cryobanking collections of viable cells.

  3. High speed flow cytometric separation of viable cells

    Science.gov (United States)

    Sasaki, Dennis T.; Van den Engh, Gerrit J.; Buckie, Anne-Marie

    1995-01-01

    Hematopoietic cell populations are separated to provide cell sets and subsets as viable cells with high purity and high yields, based on the number of original cells present in the mixture. High-speed flow cytometry is employed using light characteristics of the cells to separate the cells, where high flow speeds are used to reduce the sorting time.

  4. Surface Charge Visualization at Viable Living Cells.

    Science.gov (United States)

    Perry, David; Paulose Nadappuram, Binoy; Momotenko, Dmitry; Voyias, Philip D; Page, Ashley; Tripathi, Gyanendra; Frenguelli, Bruno G; Unwin, Patrick R

    2016-03-09

    Scanning ion conductance microscopy (SICM) is demonstrated to be a powerful technique for quantitative nanoscale surface charge mapping of living cells. Utilizing a bias modulated (BM) scheme, in which the potential between a quasi-reference counter electrode (QRCE) in an electrolyte-filled nanopipette and a QRCE in bulk solution is modulated, it is shown that both the cell topography and the surface charge present at cellular interfaces can be measured simultaneously at high spatial resolution with dynamic potential measurements. Surface charge is elucidated by probing the properties of the diffuse double layer (DDL) at the cellular interface, and the technique is sensitive at both low-ionic strength and under typical physiological (high-ionic strength) conditions. The combination of experiments that incorporate pixel-level self-referencing (calibration) with a robust theoretical model allows for the analysis of local surface charge variations across cellular interfaces, as demonstrated on two important living systems. First, charge mapping at Zea mays root hairs shows that there is a high negative surface charge at the tip of the cell. Second, it is shown that there are distinct surface charge distributions across the surface of human adipocyte cells, whose role is the storage and regulation of lipids in mammalian systems. These are new features, not previously recognized, and their implications for the functioning of these cells are highlighted.

  5. Inkjet printing of viable human dental follicle stem cells

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    Mau Robert

    2015-09-01

    Full Text Available Inkjet printing technology has the potential to be used for seeding of viable cells for tissue engineering approaches. For this reason, a piezoelectrically actuated, drop-on-demand inkjet printing system was applied to deliver viable human dental follicle stem cells (hDFSC of sizes of about 15 μm up to 20 μm in diameter. The purpose of these investigations was to verify the stability of the printing process and to evaluate cell viability post printing. Using a Nanoplotter 2.1 (Gesim, Germany equipped with the piezoelectric printhead NanoTip HV (Gesim, Germany, a concentration of 6.6 ×106 cells ml−1 in DMEM with 10% fetal calf serum (FCS could be dispensed. The piezoelectric printhead has a nominal droplet volume of ~ 400 pl and was set to a voltage of 75 V and a pulse of 50 μs while dosing 50 000 droplets over a time of 100 seconds. The volume and trajectory of the droplet were checked by a stroboscope test right before and after the printing process. It was found that the droplet volume decreases significantly by 35% during printing process, while the trajectory of the droplets remains stable with only an insignificant number of degrees deviation from the vertical line. It is highly probable that some cell sedimentations or agglomerations affect the printing performance. The cell viability post printing was assessed by using the Trypan Blue dye exclusion test. The printing process was found to have no significant influence on cell survival. In conclusion, drop-on-demand inkjet printing can be a potent tool for the seeding of viable cells.

  6. Sensitive and Specific Biomimetic Lipid Coated Microfluidics to Isolate Viable Circulating Tumor Cells and Microemboli for Cancer Detection.

    Directory of Open Access Journals (Sweden)

    Jia-Yang Chen

    Full Text Available Here we presented a simple and effective membrane mimetic microfluidic device with antibody conjugated supported lipid bilayer (SLB "smart coating" to capture viable circulating tumor cells (CTCs and circulating tumor microemboli (CTM directly from whole blood of all stage clinical cancer patients. The non-covalently bound SLB was able to promote dynamic clustering of lipid-tethered antibodies to CTC antigens and minimized non-specific blood cells retention through its non-fouling nature. A gentle flow further flushed away loosely-bound blood cells to achieve high purity of CTCs, and a stream of air foam injected disintegrate the SLB assemblies to release intact and viable CTCs from the chip. Human blood spiked cancer cell line test showed the ~95% overall efficiency to recover both CTCs and CTMs. Live/dead assay showed that at least 86% of recovered cells maintain viability. By using 2 mL of peripheral blood, the CTCs and CTMs counts of 63 healthy and colorectal cancer donors were positively correlated with the cancer progression. In summary, a simple and effective strategy utilizing biomimetic principle was developed to retrieve viable CTCs for enumeration, molecular analysis, as well as ex vivo culture over weeks. Due to the high sensitivity and specificity, it is the first time to show the high detection rates and quantity of CTCs in non-metastatic cancer patients. This work offers the values in both early cancer detection and prognosis of CTC and provides an accurate non-invasive strategy for routine clinical investigation on CTCs.

  7. The molecularly crowded cytoplasm of bacterialcCells : Dividing cells contrasted with viable but non-culturable (VBNC) bacterial cells

    NARCIS (Netherlands)

    Trevors, J. T.; van Elsas, J. D.; Bej, A. K.

    2013-01-01

    In this perspective, we discuss the cytoplasm in actively growing bacterial cells contrasted with viable but non-culturable (VBNC) cells. Actively growing bacterial cells contain a more molecularly crowded and organized cytoplasm, and are capable of completing their cell cycle resulting in cell

  8. Non-viable antagonist cells are associated with reduced biocontrol performance by viable cells of the yeast Papiliotrema flavescens against Fusarium head blight of wheat.

    Science.gov (United States)

    Microbially-based plant disease control products have achieved commercial market success, but the efficacy of such biocontrol products is sometimes deemed inconsistent. Improper processing of harvested microbial biomass or long-term storage can reduce the proportion of viable cells and necessitate t...

  9. Modelling the number of viable vegetative cells of Bacillus cereus passing through the stomach

    NARCIS (Netherlands)

    Wijnands, L.M.; Pielaat, A.; Dufrenne, J.B.; Zwietering, M.H.; Leusden, van F.M.

    2009-01-01

    Aims: Model the number of viable vegetative cells of B. cereus surviving the gastric passage after experiments in simulated gastric conditions. Materials and Methods: The inactivation of stationary and exponential phase vegetative cells of twelve different strains of Bacillus cereus, both mesophilic

  10. Combining ethidium monoazide treatment with real-time PCR selectively quantifies viable Batrachochytrium dendrobatidis cells.

    Science.gov (United States)

    Blooi, Mark; Martel, An; Vercammen, Francis; Pasmans, Frank

    2013-02-01

    Detection of the lethal amphibian fungus Batrachochytrium dendrobatidis relies on PCR-based techniques. Although highly accurate and sensitive, these methods fail to distinguish between viable and dead cells. In this study a novel approach combining the DNA intercalating dye ethidium monoazide (EMA) and real-time PCR is presented that allows quantification of viable B. dendrobatidis cells without the need for culturing. The developed method is able to suppress real-time PCR signals of heat-killed B. dendrobatidis zoospores by 99.9 % and is able to discriminate viable from heat-killed B. dendrobatidis zoospores in mixed samples. Furthermore, the novel approach was applied to assess the antifungal activity of the veterinary antiseptic F10(®) Antiseptic Solution. This disinfectant killed B. dendrobatidis zoospores effectively within 1 min at concentrations as low as 1:6400. Copyright © 2013 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  11. A simple way to identify non-viable cells within living plant tissue using confocal microscopy

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    Truernit Elisabeth

    2008-06-01

    Full Text Available Abstract Background Plant cell death is a normal process during plant development. Mutant plants may exhibit misregulation of this process, which can lead to severe growth defects. Simple ways of visualising cell death in living plant tissues can aid the study of plant development and physiology. Results Spectral variants of the fluorescent SYTOX dyes were tested for their usefulness for the detection of non-viable cells within plant embryos and roots using confocal laser-scanning microscopy. The dyes were selective for non-viable cells and showed very little background staining in living cells. Simultaneous detection of SYTOX dye and fluorescent protein (e.g. GFP fluorescence was possible. Conclusion The fluorescent SYTOX dyes are useful for an easy and quick first assay of plant cell viability in living plant samples using fluorescence and confocal laser-scanning microscopy.

  12. Separable Bilayer Microfiltration Device for Viable Label-free Enrichment of Circulating Tumour Cells

    Science.gov (United States)

    Zhou, Ming-Da; Hao, Sijie; Williams, Anthony J.; Harouaka, Ramdane A.; Schrand, Brett; Rawal, Siddarth; Ao, Zheng; Brennaman, Randall; Gilboa, Eli; Lu, Bo; Wang, Shuwen; Zhu, Jiyue; Datar, Ram; Cote, Richard; Tai, Yu-Chong; Zheng, Si-Yang

    2014-12-01

    The analysis of circulating tumour cells (CTCs) in cancer patients could provide important information for therapeutic management. Enrichment of viable CTCs could permit performance of functional analyses on CTCs to broaden understanding of metastatic disease. However, this has not been widely accomplished. Addressing this challenge, we present a separable bilayer (SB) microfilter for viable size-based CTC capture. Unlike other single-layer CTC microfilters, the precise gap between the two layers and the architecture of pore alignment result in drastic reduction in mechanical stress on CTCs, capturing them viably. Using multiple cancer cell lines spiked in healthy donor blood, the SB microfilter demonstrated high capture efficiency (78-83%), high retention of cell viability (71-74%), high tumour cell enrichment against leukocytes (1.7-2 × 103), and widespread ability to establish cultures post-capture (100% of cell lines tested). In a metastatic mouse model, SB microfilters successfully enriched viable mouse CTCs from 0.4-0.6 mL whole mouse blood samples and established in vitro cultures for further genetic and functional analysis. Our preliminary studies reflect the efficacy of the SB microfilter device to efficiently and reliably enrich viable CTCs in animal model studies, constituting an exciting technology for new insights in cancer research.

  13. Use of propidium monoazide for selective profiling of viable microbial cells during Gouda cheese ripening

    NARCIS (Netherlands)

    Erkus, O.; Jager, V.C. de; Geene, R.T.; Alen-Boerrigter, I.J. van; Hazelwood, L.; Hijum, S.A.F.T. van; Kleerebezem, M; Smid, E.J.

    2016-01-01

    DNA based microbial community profiling of food samples is confounded by the presence of DNA derived from membrane compromised (dead or injured) cells. Selective amplification of DNA from viable (intact) fraction of the community by propidium monoazide (PMA) treatment could circumvent this problem.

  14. In vitro and in vivo bioluminescent quantification of viable stem cells in engineered constructs.

    Science.gov (United States)

    Logeart-Avramoglou, Delphine; Oudina, Karim; Bourguignon, Marianne; Delpierre, Laetitia; Nicola, Marie-Anne; Bensidhoum, Morad; Arnaud, Eric; Petite, Herve

    2010-06-01

    Bioluminescent quantification of viable cells inside three-dimensional porous scaffolds was performed in vitro and in vivo. The assay quantified the bioluminescence of murine stem (C3H10T1/2) cells tagged with the luciferase gene reporter and distributed inside scaffolds of either soft, translucent, AN69 polymeric hydrogel or hard, opaque, coral ceramic materials. Quantitative evaluation of bioluminescence emitted from tagged cells adhering to these scaffolds was performed in situ using either cell lysates and a luminometer or intact cells and a bioluminescence imaging system. Despite attenuation of the signal when compared to cells alone, the bioluminescence correlated with the number of cells (up to 1.5 x 10(5)) present on each material scaffold tested, both in vitro and noninvasively in vivo (subcutaneous implants in the mouse model). The noninvasive bioluminescence measurement technique proved to be comparable to the cell-destructive bioluminescence measurement technique. Monitoring the kinetics of luciferase expression via bioluminescence enabled real-time assessment of cell survival and proliferation on the scaffolds tested over prolonged (up to 59 days) periods of time. This novel, sensitive, easy, fast-to-implement, quantitative bioluminescence assay has great, though untapped, potential for screening and determining noninvasively the presence of viable cells on biomaterial constructs in the tissue engineering and tissue regeneration fields.

  15. A PCR-Based Method for Monitoring Legionella pneumophila in Water Samples Detects Viable but Noncultivable Legionellae That Can Recover Their Cultivability▿

    Science.gov (United States)

    Dusserre, Eric; Ginevra, Christophe; Hallier-Soulier, Sylvie; Vandenesch, François; Festoc, Gabriel; Etienne, Jerome; Jarraud, Sophie; Molmeret, Maëlle

    2008-01-01

    Legionella pneumophila is the causative agent of Legionnaires' disease. This bacterium is ubiquitous in aqueous environments and uses amoebae as an intracellular replicative niche. Real-time PCR has been developed for rapid detection of Legionella DNA in water samples. In addition to culturable bacteria, this method may also detect dead and viable but noncultivable (VBNC) legionellae. In order to understand the significance of positive PCR results in this setting, we prepared water samples containing known concentrations of L. pneumophila and analyzed them comparatively by means of conventional culture, real-time PCR, viability labeling, and immunodetection (solid-phase cytometry). We also examined the influence of chlorination on the results of the four methods. The different techniques yielded similar results for nonchlorinated water samples but not for chlorinated samples. After treatment for 24 h with 0.5 and 1 ppm chlorine, all cultures were negative, PCR and immunodetection showed about 106 genome units and bacteria/ml, and total-viable-count (TVC) labeling detected 105 and 102 metabolically active bacteria/ml, respectively. Thus, PCR also detected bacteria that were VBNC. The recoverability of VBNC forms was confirmed by 5 days of coculture with Acanthamoeba polyphaga. Therefore, some TVC-positive bacteria were potentially infective. These data show that L. pneumophila PCR detects not only culturable bacteria but also VBNC forms and dead bacterial DNA at low chlorine concentrations. PMID:18515476

  16. Cancer cells recovering from damage exhibit mitochondrial restructuring and increased aerobic glycolysis

    Energy Technology Data Exchange (ETDEWEB)

    Akakura, Shin; Ostrakhovitch, Elena; Sanokawa-Akakura, Reiko [Frontiers in Bioscience Research Institute in Aging and Cancer, University of California, Irvine, CA (United States); Tabibzadeh, Siamak, E-mail: fbs@bioscience.org [Frontiers in Bioscience Research Institute in Aging and Cancer, University of California, Irvine, CA (United States); Dept of Oncologic Radiology, University of California, Irvine, CA (United States)

    2014-06-13

    Highlights: • Some cancer cells recover from severe damage that causes cell death in majority of cells. • Damage-Recovered (DR) cancer cells show reduced mitochondria, mDNA and mitochondrial enzymes. • DR cells show increased aerobic glycolysis, ATP, cell proliferation, and resistance to damage. • DR cells recovered from in vivo damage also show increased glycolysis and proliferation rate. - Abstract: Instead of relying on mitochondrial oxidative phosphorylation, most cancer cells rely heavily on aerobic glycolysis, a phenomenon termed as “the Warburg effect”. We considered that this effect is a direct consequence of damage which persists in cancer cells that recover from damage. To this end, we studied glycolysis and rate of cell proliferation in cancer cells that recovered from severe damage. We show that in vitro Damage-Recovered (DR) cells exhibit mitochondrial structural remodeling, display Warburg effect, and show increased in vitro and in vivo proliferation and tolerance to damage. To test whether cancer cells derived from tumor microenvironment can show similar properties, we isolated Damage-Recovered (T{sup DR}) cells from tumors. We demonstrate that T{sup DR} cells also show increased aerobic glycolysis and a high proliferation rate. These findings show that Warburg effect and its consequences are induced in cancer cells that survive severe damage.

  17. Mobilization of Viable Tumor Cells Into the Circulation During Radiation Therapy

    Energy Technology Data Exchange (ETDEWEB)

    Martin, Olga A. [Division of Radiation Oncology and Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Molecular Radiation Biology Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); The Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, VIC (Australia); Anderson, Robin L. [The Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, VIC (Australia); Metastasis Research Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Russell, Prudence A. [Department of Anatomical Pathology, St. Vincent Hospital, Fitzroy, VIC (Australia); Ashley Cox, R. [Division of Radiation Oncology and Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Ivashkevich, Alesia [Molecular Radiation Biology Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Laboratory of DNA Repair and Genomics, Centre for Innate Immunity and Infectious Disease, Monash Institute for Medical Research, Monash University, Clayton, VIC (Australia); Swierczak, Agnieszka; Doherty, Judy P. [Metastasis Research Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Jacobs, Daphne H.M. [Division of Radiation Oncology and Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Smith, Jai [Molecular Radiation Biology Laboratory, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Siva, Shankar; Daly, Patricia E. [Division of Radiation Oncology and Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); Ball, David L. [Division of Radiation Oncology and Cancer Imaging, Peter MacCallum Cancer Centre, East Melbourne, VIC (Australia); The Sir Peter MacCallum Department of Oncology, University of Melbourne, Melbourne, VIC (Australia); and others

    2014-02-01

    Purpose: To determine whether radiation therapy (RT) could mobilize viable tumor cells into the circulation of non-small cell lung cancer (NSCLC) patients. Methods and Materials: We enumerated circulating tumor cells (CTCs) by fluorescence microscopy of blood samples immunostained with conventional CTC markers. We measured their DNA damage levels using γ-H2AX, a biomarker for radiation-induced DNA double-strand breaks, either by fluorescence-activated cell sorting or by immunofluorescence microscopy. Results: Twenty-seven RT-treated NSCLC patients had blood samples analyzed by 1 or more methods. We identified increased CTC numbers after commencement of RT in 7 of 9 patients treated with palliative RT, and in 4 of 8 patients treated with curative-intent RT. Circulating tumor cells were also identified, singly and in clumps in large numbers, during RT by cytopathologic examination (in all 5 cases studied). Elevated γ-H2AX signal in post-RT blood samples signified the presence of CTCs derived from irradiated tumors. Blood taken after the commencement of RT contained tumor cells that proliferated extensively in vitro (in all 6 cases studied). Circulating tumor cells formed γ-H2AX foci in response to ex vivo irradiation, providing further evidence of their viability. Conclusions: Our findings provide a rationale for the development of strategies to reduce the concentration of viable CTCs by modulating RT fractionation or by coadministering systemic therapies.

  18. Desiccation induces viable but Non-Culturable cells in Sinorhizobium meliloti 1021.

    Science.gov (United States)

    Vriezen, Jan Ac; de Bruijn, Frans J; Nüsslein, Klaus R

    2012-01-20

    Sinorhizobium meliloti is a microorganism commercially used in the production of e.g. Medicago sativa seed inocula. Many inocula are powder-based and production includes a drying step. Although S. meliloti survives drying well, the quality of the inocula is reduced during this process. In this study we determined survival during desiccation of the commercial strains 102F84 and 102F85 as well as the model strain USDA1021.The survival of S. meliloti 1021 was estimated during nine weeks at 22% relative humidity. We found that after an initial rapid decline of colony forming units, the decline slowed to a steady 10-fold reduction in colony forming units every 22 days. In spite of the reduction in colony forming units, the fraction of the population identified as viable (42-54%) based on the Baclight live/dead stain did not change significantly over time. This change in the ability of viable cells to form colonies shows (i) an underestimation of the survival of rhizobial cells using plating methods, and that (ii) in a part of the population desiccation induces a Viable But Non Culturable (VBNC)-like state, which has not been reported before. Resuscitation attempts did not lead to a higher recovery of colony forming units indicating the VBNC state is stable under the conditions tested. This observation has important consequences for the use of rhizobia. Finding methods to resuscitate this fraction may increase the quality of powder-based seed inocula.

  19. Mathematical modelling of the viable epidermis: impact of cell shape and vertical arrangement

    KAUST Repository

    Wittum, Rebecca

    2017-12-07

    In-silico methods are valuable tools for understanding the barrier function of the skin. The key benefit is that mathematical modelling allows the interplay between cell shape and function to be elucidated. This study focuses on the viable (living) epidermis. For this region, previous works suggested a diffusion model and an approximation of the cells by hexagonal prisms. The work at hand extends this in three ways. First, the extracellular space is treated with full spatial resolution. This induces a decrease of permeability by about 10%. Second, cells of tetrakaidecahedral shape are considered, in addition to the original hexagonal prisms. For both cell types, the resulting membrane permeabilities are compared. Third, for the first time, the influence of cell stacking in the vertical direction is considered. This is particularly important for the stratum granulosum, where tight junctions are present.

  20. Progress in biocatalysis with immobilized viable whole cells: systems development, reaction engineering and applications.

    Science.gov (United States)

    Polakovič, Milan; Švitel, Juraj; Bučko, Marek; Filip, Jaroslav; Neděla, Vilém; Ansorge-Schumacher, Marion B; Gemeiner, Peter

    2017-05-01

    Viable microbial cells are important biocatalysts in the production of fine chemicals and biofuels, in environmental applications and also in emerging applications such as biosensors or medicine. Their increasing significance is driven mainly by the intensive development of high performance recombinant strains supplying multienzyme cascade reaction pathways, and by advances in preservation of the native state and stability of whole-cell biocatalysts throughout their application. In many cases, the stability and performance of whole-cell biocatalysts can be highly improved by controlled immobilization techniques. This review summarizes the current progress in the development of immobilized whole-cell biocatalysts, the immobilization methods as well as in the bioreaction engineering aspects and economical aspects of their biocatalytic applications.

  1. Trypan blue dye enters viable cells incubated with the pore-forming toxin HlyII of Bacillus cereus.

    Directory of Open Access Journals (Sweden)

    Seav-Ly Tran

    Full Text Available Trypan blue is a dye that has been widely used for selective staining of dead tissues or cells. Here, we show that the pore-forming toxin HlyII of Bacillus cereus allows trypan blue staining of macrophage cells, despite the cells remaining viable and metabolically active. These findings suggest that the dye enters viable cells through the pores. To our knowledge, this is the first demonstration that trypan blue may enter viable cells. Consequently, the use of trypan blue staining as a marker of vital status should be interpreted with caution. The blue coloration does not necessarily indicate cell lysis, but may rather indicate pore formation in the cell membranes and more generally increased membrane permeability.

  2. IN-VITRO BIOREDUCTION OF HEXAVALENT CHROMIUM BY VIABLE WHOLE CELLS OF Arthrobacter sp. SUK 1201

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    Satarupa Dey

    2014-08-01

    Full Text Available A chromium resistant and reducing bacterium Arthrobacter sp. SUK 1201 was isolated from chromite mine overburden dumps of Orissa, India. Viable whole cells of this isolate was capable of completely reducing 100 µM Cr(VI in chemically defined MS medium within 28 h of incubation under batch cultivation. Reduction of chromate increased with increased cell density and was maximum at a density of 1010 cells/ml, but the reduction potential of the suspended cells decreased with increase in Cr(VI concentration in the medium. Chromate reducing efficiency was promoted when glycerol and glucose was used as electron donors, while the optimum pH and temperature of Cr(VI reduction was found to be 7.0 and 35°C respectively. The reduction process was inhibited by divalent cations Ni, Co and Cd, but not by Cu and Fe. Similarly, carbonyl cyanide m-chlorophenylhydrazone (CCCP, N,N,-Di cyclohexyl carboiimide (DCC, sodium azide and sodium fluoride were inhibitory to chromate reduction, while in presence of 2,4 dinitrophenol (2,4 DNP chromate reduction by SUK 1201 cells remained unaffected.

  3. Viable bacteria associated with red blood cells and plasma in freshly drawn blood donations.

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    Christian Damgaard

    Full Text Available Infection remains a leading cause of post-transfusion mortality and morbidity. Bacterial contamination is, however, detected in less than 0.1% of blood units tested. The aim of the study was to identify viable bacteria in standard blood-pack units, with particular focus on bacteria from the oral cavity, and to determine the distribution of bacteria revealed in plasma and in the red blood cell (RBC-fraction.Cross-sectional study. Blood were separated into plasma and RBC-suspensions, which were incubated anaerobically or aerobically for 7 days on trypticase soy blood agar (TSA or blue lactose plates. For identification colony PCR was performed using primers targeting 16S rDNA.Blood donors attending Capital Region Blood Bank, Copenhagen University Hospital, Rigshospitalet, Hvidovre, Denmark, October 29th to December 10th 2013.60 donors (≥50 years old, self-reported medically healthy.Bacterial growth was observed on plates inoculated with plasma or RBCs from 62% of the blood donations. Growth was evident in 21 (35% of 60 RBC-fractions and in 32 (53% of 60 plasma-fractions versus 8 of 60 negative controls (p = 0.005 and p = 2.6x10-6, respectively. Propionibacterium acnes was found in 23% of the donations, and Staphylococcus epidermidis in 38%. The majority of bacteria identified in the present study were either facultative anaerobic (59.5% or anaerobic (27.8% species, which are not likely to be detected during current routine screening.Viable bacteria are present in blood from donors self-reported as medically healthy, indicating that conventional test systems employed by blood banks insufficiently detect bacteria in plasma. Further investigation is needed to determine whether routine testing for anaerobic bacteria and testing of RBC-fractions for adherent bacteria should be recommended.

  4. Increasing Vero viable cell densities for yellow fever virus production in stirred-tank bioreactors using serum-free medium.

    Science.gov (United States)

    Mattos, Diogo A; Silva, Marlon V; Gaspar, Luciane P; Castilho, Leda R

    2015-08-20

    In this work, changes in Vero cell cultivation methods have been employed in order to improve cell growth conditions to obtain higher viable cell densities and to increase viral titers. The propagation of the 17DD yellow fever virus (YFV) in Vero cells grown on Cytodex I microcarriers was evaluated in 3-L bioreactor vessels. Prior to the current changes, Vero cells were repeatedly displaying insufficient microcarrier colonization. A modified cultivation process with four changes has resulted in higher cell densities and higher virus titers than previously observed for 17DD YFV. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. Cellular bone matrices: viable stem cell-containing bone graft substitutes.

    Science.gov (United States)

    Skovrlj, Branko; Guzman, Javier Z; Al Maaieh, Motasem; Cho, Samuel K; Iatridis, James C; Qureshi, Sheeraz A

    2014-11-01

    Advances in the field of stem cell technology have stimulated the development and increased use of allogenic bone grafts containing live mesenchymal stem cells (MSCs), also known as cellular bone matrices (CBMs). It is estimated that CBMs comprise greater than 17% of all bone grafts and bone graft substitutes used. To critically evaluate CBMs, specifically their technical specifications, existing published data supporting their use, US Food and Drug Administration (FDA) regulation, cost, potential pitfalls, and other aspects pertaining to their use. Areview of literature. A series of Ovid, Medline, and Pubmed-National Library of Medicine/National Institutes of Health (www.ncbi.nlm.nih.gov) searches were performed. Only articles in English journals or published with English language translations were included. Level of evidence of the selected articles was assessed. Specific technical information on each CBM was obtained by direct communication from the companies marketing the individual products. Five different CBMs are currently available for use in spinal fusion surgery. There is a wide variation between the products with regard to the average donor age at harvest, total cellular concentration, percentage of MSCs, shelf life, and cell viability after defrosting. Three retrospective studies evaluating CBMs and fusion have shown fusion rates ranging from 90.2% to 92.3%, and multiple industry-sponsored trials are underway. No independent studies evaluating spinal fusion rates with the use of CBMs exist. All the commercially available CBMs claim to meet the FDA criteria under Section 361, 21 CFR Part 1271, and are not undergoing FDA premarket review. The CBMs claim to provide viable MSCs and are offered at a premium cost. Numerous challenges exist in regard to MSCs' survival, function, osteoblastic potential, and cytokine production once implanted into the intended host. Cellular bone matrices may be a promising bone augmentation technology in spinal fusion surgery

  6. DEMONSTRATION OF FUEL CELLS TO RECOVER ENERGY FROM LANDFILL GAS: PHASE II. PRETREATMENT SYSTEM PERFORMANCE MEASUREMENT

    Science.gov (United States)

    The report describes Phase II of a demonstration of the utilization of commercial phosphoric acid fuel cells to recover energy from landfill gas. This phase consisted primarily of the construction and testing of a Gas Pretreatment Unit (GPU) whose function is to remove those impu...

  7. Combined bromodeoxyuridine immunohistochemistry and Masson trichrome staining: facilitated detection of cell proliferation in viable vs. infarcted myocardium.

    Science.gov (United States)

    Lazarous, D F; Shou, M; Unger, E F

    1992-09-01

    Cells in the S-phase of the cell cycle can be identified in tissue sections by immunohistochemical localization of the thymidine analogue bromodeoxyuridine (BrdU). Generally, a single counterstain is used to visualize the underlying tissue; however, interpretation of morphologic detail is often difficult. We have utilized BrdU to localize proliferating cells in myocardium exposed to angiogenic mitogens. To facilitate identification of labelled nuclei in the context of infarcted vs. viable myocardium, BrdU immunohistochemistry was followed by a modified Masson trichrome stain. The time of exposure to the counterstains and the wash protocol were re-revised, permitting clear identification of the labelled brown nuclei against a background of red viable myocardium vs. blue infarct. The combined technique also provides color contrast suitable for computer-based image analysis.

  8. Quantitative assessment of viable cells of Lactobacillus plantarum strains in single, dual and multi-strain biofilms.

    Science.gov (United States)

    Fernández Ramírez, Mónica D; Kostopoulos, Ioannis; Smid, Eddy J; Nierop Groot, Masja N; Abee, Tjakko

    2017-03-06

    Biofilms of Lactobacillus plantarum are a potential source for contamination and recontamination of food products. Although biofilms have been mostly studied using single species or even single strains, it is conceivable that in a range of environmental settings including food processing areas, biofilms are composed of multiple species with each species represented by multiple strains. In this study six spoilage related L. plantarum strains FBR1-FBR6 and the model strain L. plantarum WCFS1 were characterised in single, dual and multiple strain competition models. A quantitative PCR approach was used with added propidium monoazide (PMA) enabling quantification of intact cells in the biofilm, representing the viable cell fraction that determines the food spoilage risk. Our results show that the performance of individual strains in multi-strain cultures generally correlates with their performance in pure culture, and relative strain abundance in multi-strain biofilms positively correlated with the relative strain abundance in suspended (planktonic) cultures. Performance of individual strains in dual-strain biofilms was highly influenced by the presence of the secondary strain, and in most cases no correlation between the relative contributions of viable planktonic cells and viable cells in the biofilm was noted. The total biofilm quantified by CV staining of the dual and multi-strain biofilms formed was mainly correlated to CV values of the dominant strain obtained in single strain studies. However, the combination of strain FBR5 and strain WCFS1 showed significantly higher CV values compared to the individual performances of both strains indicating that total biofilm formation was higher in this specific condition. Notably, L. plantarum FBR5 was able to outgrow all other strains and showed the highest relative abundance in dual and multi-strain biofilms. All the dual and multi-strain biofilms contained a considerable number of viable cells, representing a potential

  9. A multimodality imaging model to track viable breast cancer cells from single arrest to metastasis in the mouse brain

    Science.gov (United States)

    Parkins, Katie M.; Hamilton, Amanda M.; Makela, Ashley V.; Chen, Yuanxin; Foster, Paula J.; Ronald, John A.

    2016-10-01

    Cellular MRI involves sensitive visualization of iron-labeled cells in vivo but cannot differentiate between dead and viable cells. Bioluminescence imaging (BLI) measures cellular viability, and thus we explored combining these tools to provide a more holistic view of metastatic cancer cell fate in mice. Human breast carcinoma cells stably expressing Firefly luciferase were loaded with iron particles, injected into the left ventricle, and BLI and MRI were performed on days 0, 8, 21 and 28. The number of brain MR signal voids (i.e., iron-loaded cells) on day 0 significantly correlated with BLI signal. Both BLI and MRI signals decreased from day 0 to day 8, indicating a loss of viable cells rather than a loss of iron label. Total brain MR tumour volume on day 28 also correlated with BLI signal. Overall, BLI complemented our sensitive cellular MRI technologies well, allowing us for the first time to screen animals for successful injections, and, in addition to MR measures of cell arrest and tumor burden, provided longitudinal measures of cancer cell viability in individual animals. We predict this novel multimodality molecular imaging framework will be useful for evaluating the efficacy of emerging anti-cancer drugs at different stages of the metastatic cascade.

  10. Viable cell yield from active dry yeast products and effects of storage temperature and diluent on yeast cell viability.

    Science.gov (United States)

    Sullivan, M L; Bradford, B J

    2011-01-01

    Active dry yeast (ADY) products are commonly fed in the dairy industry, but research regarding quality control for such products is limited. The objectives of this study were to determine yeast viability in field samples relative to manufacturers' guarantees (experiment 1), measure the effects of high-temperature storage on yeast viability (experiment 1), and determine the effect of vitamin-trace mineral (VTM) premix on yeast viability (experiment 2). Commercially available ADY products were acquired in triplicate through normal distribution channels and stored at 4°C upon receipt. Initial samples were evaluated for colony-forming units and compared with product label guarantees. Only 1 of the 6 products sampled in experiment 1 met product guarantees for all 3 samples. To determine effects of storage temperature and duration on viability, ADY samples were stored in an incubator at 40°C with ambient humidity for 1, 2, and 3 mo. High-temperature storage significantly decreased viability over the 3-mo period; approximately 90% of viable cells were lost each month. Three of the 5 products sampled in experiment 2 met product guarantees. Fresh samples of 4 of these 5 ADY products were mixed in duplicate with ground corn (GC) or a VTM premix to achieve a target concentration of 2.2×10(8) cfu/g. For each product, GC and VTM samples were stored at ambient temperature (22°C) and at an elevated temperature (40°C) for 2 wk. No differences in viable yeast count were observed between GC and VTM samples immediately after mixing or after storage at ambient temperature. Yeast viability in GC and VTM samples decreased during storage at an elevated temperature. There also was a significant interaction of diluent and storage temperature; VTM samples had higher cell viability than GC samples when subjected to high-temperature storage. Results suggest that (1) ADY products failed to consistently meet product guarantees; (2) viability of ADY products was greatly diminished during

  11. Heritable Genetic Changes in Cells Recovered From Irradiated 3D Tissue Constructs

    Energy Technology Data Exchange (ETDEWEB)

    Michael Cornforth

    2012-03-26

    Combining contemporary cytogenetic methods with DNA CGH microarray technology and chromosome flow-sorting increases substantially the ability to resolve exchange breakpoints associated with interstitial deletions and translocations, allowing the consequences of radiation damage to be directly measured at low doses, while also providing valuable insights into molecular mechanisms of misrepair processes that, in turn, identify appropriate biophysical models of risk at low doses. Specific aims apply to cells recovered from 3D tissue constructs of human skin and, for the purpose of comparison, the same cells irradiated in traditional 2D cultures. The project includes research complementary to NASA/HRP space radiation project.

  12. Viable cell sorting of dinoflagellates by multi-parametric flow cytometry.

    Science.gov (United States)

    Electronic cell sorting for isolation and culture of dinoflagellates and other marine eukaryotic phytoplankton was compared to the traditional method of manually picking of cells using a micropipette. Trauma to electronically sorted cells was not a limiting factor as fragile dinoflagellates, such a...

  13. Method for recovering catalytic elements from fuel cell membrane electrode assemblies

    Science.gov (United States)

    Shore, Lawrence [Edison, NJ; Matlin, Ramail [Berkeley Heights, NJ; Heinz, Robert [Ludwigshafen, DE

    2012-06-26

    A method for recovering catalytic elements from a fuel cell membrane electrode assembly is provided. The method includes converting the membrane electrode assembly into a particulate material, wetting the particulate material, forming a slurry comprising the wetted particulate material and an acid leachate adapted to dissolve at least one of the catalytic elements into a soluble catalytic element salt, separating the slurry into a depleted particulate material and a supernatant containing the catalytic element salt, and washing the depleted particulate material to remove any catalytic element salt retained within pores in the depleted particulate material.

  14. Revisiting the single cell protein application of Cupriavidus necator H16 and recovering bioplastic granules simultaneously.

    Science.gov (United States)

    Kunasundari, Balakrishnan; Murugaiyah, Vikneswaran; Kaur, Gurjeet; Maurer, Frans H J; Sudesh, Kumar

    2013-01-01

    Cupriavidus necator H16 (formerly known as Hydrogenomonas eutropha) was famous as a potential single cell protein (SCP) in the 1970s. The drawback however was the undesirably efficient accumulation of non-nutritive polyhydroxybutyrate (PHB) storage compound in the cytoplasm of this bacterium. Eventually, competition from soy-based protein resulted in SCP not receiving much attention. Nevertheless, C. necator H16 remained in the limelight as a producer of PHB, which is a material that resembles commodity plastics such as polypropylene. PHB is a 100% biobased and biodegradable polyester. Although tremendous achievements have been attained in the past 3 decades in the efficient production of PHB, this bioplastic is still costly. One of the main problems has been the recovery of PHB from the cell cytoplasm. In this study, we showed for the first time that kilogram quantities of PHB can be easily recovered in the laboratory without the use of any solvents and chemicals, just by using the cells as SCP. In addition, the present study also demonstrated the safety and tolerability of animal model used, Sprague Dawley given lyophilized cells of C. necator H16. The test animals readily produced fecal pellets that were whitish in color, as would be expected of PHB granules. The pellets were determined to contain about 82-97 wt% PHB and possessed molecular mass of around 930 kg/mol. The PHB granules recovered biologically possessed similar molecular mass compared to chloroform extracted PHB [950 kg/mol]. This method now allows the production and purification of substantial quantities of PHB for various experimental trials. The method reported here is easy, does not require expensive instrumentation, scalable and does not involve extensive use of solvents and strong chemicals.

  15. Revisiting the single cell protein application of Cupriavidus necator H16 and recovering bioplastic granules simultaneously.

    Directory of Open Access Journals (Sweden)

    Balakrishnan Kunasundari

    Full Text Available Cupriavidus necator H16 (formerly known as Hydrogenomonas eutropha was famous as a potential single cell protein (SCP in the 1970s. The drawback however was the undesirably efficient accumulation of non-nutritive polyhydroxybutyrate (PHB storage compound in the cytoplasm of this bacterium. Eventually, competition from soy-based protein resulted in SCP not receiving much attention. Nevertheless, C. necator H16 remained in the limelight as a producer of PHB, which is a material that resembles commodity plastics such as polypropylene. PHB is a 100% biobased and biodegradable polyester. Although tremendous achievements have been attained in the past 3 decades in the efficient production of PHB, this bioplastic is still costly. One of the main problems has been the recovery of PHB from the cell cytoplasm. In this study, we showed for the first time that kilogram quantities of PHB can be easily recovered in the laboratory without the use of any solvents and chemicals, just by using the cells as SCP. In addition, the present study also demonstrated the safety and tolerability of animal model used, Sprague Dawley given lyophilized cells of C. necator H16. The test animals readily produced fecal pellets that were whitish in color, as would be expected of PHB granules. The pellets were determined to contain about 82-97 wt% PHB and possessed molecular mass of around 930 kg/mol. The PHB granules recovered biologically possessed similar molecular mass compared to chloroform extracted PHB [950 kg/mol]. This method now allows the production and purification of substantial quantities of PHB for various experimental trials. The method reported here is easy, does not require expensive instrumentation, scalable and does not involve extensive use of solvents and strong chemicals.

  16. Fuel cells are a commercially viable alternative for the production of "clean" energy.

    Science.gov (United States)

    Niakolas, Dimitris K; Daletou, Maria; Neophytides, Stylianos G; Vayenas, Constantinos G

    2016-01-01

    Fuel cells present a highly efficient and environmentally friendly alternative technology for decentralized energy production. The scope of the present study is to provide an overview of the technological and commercialization readiness level of fuel cells. Specifically, there is a brief description of their general advantages and weaknesses in correlation with various technological actions and political strategies, which are adopted towards their proper positioning in the global market. Some of the most important key performance indicators are also discussed, alongside with a few examples of broad commercialization. It is concluded that the increasing number of companies which utilize and invest on this technology, in combination with the supply chain improvements and the concomitant technological maturity and recognition, reinforce the fuel cell industry so as to become well-aligned for global success.

  17. A viable electrode material for use in microbial fuel cells for tropical regions

    DEFF Research Database (Denmark)

    Offei, Felix; Thygesen, Anders; Mensah, Moses

    2016-01-01

    Electrode materials are critical for microbial fuel cells (MFC) since they influence the construction and operational costs. This study introduces a simple and efficient electrode material in the form of palm kernel shell activated carbon (AC) obtained in tropical regions. The novel introduction...

  18. Progress in emerging techniques for characterization of immobilized viable whole-cell biocatalysts

    Czech Academy of Sciences Publication Activity Database

    Bučko, M.; Vikartovská, A.; Schenkmayerová, A.; Tkáč, J.; Filip, J.; Chorvát Jr., D.; Neděla, Vilém; Ansorge-Schumacher, M.B.; Gemeiner, P.

    2017-01-01

    Roč. 71, č. 11 (2017), s. 2309-2324 ISSN 0366-6352 Institutional support: RVO:68081731 Keywords : bioelectrocatalysis * imaging techniques * immobilized whole- cell biocatalyst * multienzyme cascade reactions * online kinetics Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.258, year: 2016

  19. Progress in biocatalysis with immobilized viable whole cells: systems development, reaction engineering and applications

    Czech Academy of Sciences Publication Activity Database

    Polakovič, M.; Švitel, J.; Bučko, M.; Filip, J.; Neděla, Vilém; Ansorge-Schumacher, M.B.; Gemeiner, P.

    2017-01-01

    Roč. 39, č. 5 (2017), s. 667-683 ISSN 0141-5492 Institutional support: RVO:68081731 Keywords : biocatalysis * immobilization methods * immobilized whole-cell biocatalyst * multienzyme cascade reactions * process economics * reaction engineering Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 1.730, year: 2016

  20. Identifying viable regulatory and innovation pathways for regenerative medicine: a case study of cultured red blood cells.

    Science.gov (United States)

    Mittra, J; Tait, J; Mastroeni, M; Turner, M L; Mountford, J C; Bruce, K

    2015-01-25

    The creation of red blood cells for the blood transfusion markets represents a highly innovative application of regenerative medicine with a medium term (5-10 year) prospect for first clinical studies. This article describes a case study analysis of a project to derive red blood cells from human embryonic stem cells, including the systemic challenges arising from (i) the selection of appropriate and viable regulatory protocols and (ii) technological constraints related to stem cell manufacture and scale up to clinical Good Manufacturing Practice (GMP) standard. The method used for case study analysis (Analysis of Life Science Innovation Systems (ALSIS)) is also innovative, demonstrating a new approach to social and natural science collaboration to foresight product development pathways. Issues arising along the development pathway include cell manufacture and scale-up challenges, affected by regulatory demands emerging from the innovation ecosystem (preclinical testing and clinical trials). Our discussion reflects on the efforts being made by regulators to adapt the current pharmaceuticals-based regulatory model to an allogeneic regenerative medicine product and the broader lessons from this case study for successful innovation and translation of regenerative medicine therapies, including the role of methodological and regulatory innovation in future development in the field. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  1. Real-Time PCR Methodology for Selective Detection of Viable Escherichia coli O157:H7 Cells by Targeting Z3276 as a Genetic Marker

    Science.gov (United States)

    Chen, Jin-Qiang

    2012-01-01

    The goal of this study was to develop a sensitive, specific, and accurate method for the selective detection of viable Escherichia coli O157:H7 cells in foods. A unique open reading frame (ORF), Z3276, was identified as a specific genetic marker for the detection of E. coli O157:H7. We developed a real-time PCR assay with primers and probe targeting ORF Z3276 and confirmed that this assay was sensitive and specific for E. coli O157:H7 strains (n = 298). Using this assay, we can detect amounts of genomic DNA of E. coli O157:H7 as low as a few CFU equivalents. Moreover, we have developed a new propidium monoazide (PMA)–real-time PCR protocol that allows for the clear differentiation of viable from dead cells. In addition, the protocol was adapted to a 96-well plate format for easy and consistent handling of a large number of samples. Amplification of DNA from PMA-treated dead cells was almost completely inhibited, in contrast to the virtually unaffected amplification of DNA from PMA-treated viable cells. With beef spiked simultaneously with 8 × 107 dead cells/g and 80 CFU viable cells/g, we were able to selectively detect viable E. coli O157:H7 cells with an 8-h enrichment. In conclusion, this PMA–real-time PCR assay offers a sensitive and specific means to selectively detect viable E. coli O157:H7 cells in spiked beef. It also has the potential for high-throughput selective detection of viable E. coli O157:H7 cells in other food matrices and, thus, will have an impact on the accurate microbiological and epidemiological monitoring of food safety and environmental sources. PMID:22635992

  2. Viable Bacteria Associated with Red Blood Cells and Plasma in Freshly Drawn Blood Donations

    DEFF Research Database (Denmark)

    Damgaard, Christian; Magnussen, Karin; Enevold, Christian

    2015-01-01

    the oral cavity, and to determine the distribution of bacteria revealed in plasma and in the red blood cell (RBC)-fraction. DESIGN: Cross-sectional study. Blood were separated into plasma and RBC-suspensions, which were incubated anaerobically or aerobically for 7 days on trypticase soy blood agar (TSA......), self-reported medically healthy. RESULTS: Bacterial growth was observed on plates inoculated with plasma or RBCs from 62% of the blood donations. Growth was evident in 21 (35%) of 60 RBC-fractions and in 32 (53%) of 60 plasma-fractions versus 8 of 60 negative controls (p = 0.005 and p = 2.6x10...... of RBC-fractions for adherent bacteria should be recommended....

  3. A Viable Electrode Material for Use in Microbial Fuel Cells for Tropical Regions

    Directory of Open Access Journals (Sweden)

    Felix Offei

    2016-01-01

    Full Text Available Electrode materials are critical for microbial fuel cells (MFC since they influence the construction and operational costs. This study introduces a simple and efficient electrode material in the form of palm kernel shell activated carbon (AC obtained in tropical regions. The novel introduction of this material is also targeted at introducing an inexpensive and durable electrode material, which can be produced in rural communities to improve the viability of MFCs. The maximum voltage and power density obtained (under 1000 Ω load using an H-shaped MFC with AC as both anode and cathode electrode material was 0.66 V and 1.74 W/m3, respectively. The power generated by AC was as high as 86% of the value obtained with the extensively used carbon paper. Scanning electron microscopy and Denaturing Gradient Gel Electrophoresis (DGGE analysis of AC anode biofilms confirmed that electrogenic bacteria were present on the electrode surface for substrate oxidation and the formation of nanowires.

  4. Response of Listeria monocytogenes to disinfection stress at the single-cell and population levels as monitored by intracellular pH measurements and viable-cell counts

    DEFF Research Database (Denmark)

    Kastbjerg, Vicky Gaedt; Nielsen, Dennis S.; Arneborg, Nils

    2009-01-01

    .05). The protective effect of NaCl was reflected by viable-cell counts at a higher concentration of Incimaxx (0.0031%), where the salt-grown population survived better than the population grown without NaCl (P ... that a population of L. monocytogenes cells, whether planktonic or attached, is homogenous with respect to sensitivity to an acidic disinfectant studied on the single-cell level. Hence a major subpopulation more tolerant to disinfectants, and hence more persistent, does not appear to be present....

  5. A factor converting viable but nonculturable Vibrio cholerae to a culturable state in eukaryotic cells is a human catalase.

    Science.gov (United States)

    Senoh, Mitsutoshi; Hamabata, Takashi; Takeda, Yoshifumi

    2015-08-01

    In our previous work, we demonstrated that viable but nonculturable (VBNC) Vibrio cholerae O1 and O139 were converted to culturable by coculture with eukaryotic cells. Furthermore, we isolated a factor converting VBNC V. cholerae to culturable (FCVC) from a eukaryotic cell line, HT-29. In this study, we purified FCVC by successive column chromatographies comprising UNO Q-6 anion exchange, Bio-Scale CHT2-1 hydroxyapatite, and Superdex 200 10/300 GL. Homogeneity of the purified FCVC was demonstrated by SDS-PAGE. Nano-LC MS/MS analysis showed that the purified FCVC was a human catalase. An experiment of RNAi knockdown of catalase mRNA from HT-29 cells and treatment of the purified FCVC with a catalase inhibitor, 3-amino-1,2,4-triazole confirmed that the FCVC was a catalase. A possible role of the catalase in converting a VBNC V. cholerae to a culturable state in the human intestine is discussed. © 2015 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  6. Heritable Genetic Changes in Cells Recovered From Irradiated 3D Tissue Contracts. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Cornforth, Michael N. [The University of Texas Medical Branch at Galveston, TX (United States)

    2013-05-03

    Combining contemporary cytogenetic methods with DNA CGH microarray technology and chromosome flow-sorting increases substantially the ability to resolve exchange breakpoints associated with interstitial deletions and translocations, allowing the consequences of radiation damage to be directly measured at low doses, while also providing valuable insights into molecular mechanisms of misrepair processes that, in turn, identify appropriate biophysical models of risk at low doses. The aims of this work apply to cells recovered from 3D tissue constructs of human skin and, for the purpose of comparison, the same cells irradiated in traditional 2D cultures. These aims are: to analyze by multi-flour fluorescence in situ hybridization (mFISH) the chromosomes in clonal descendents of individual human fibroblasts that were previously irradiated; to examine irradiated clones from Aim 1 for submicroscopic deletions by subjecting their DNA to comparative genomic hybridization (CGH) microarray analysis; and to flow-sort aberrant chromosomes from clones containing stable radiation-induced translocations and map the breakpoints to within an average resolution of 100 kb using the technique of 'array painting'.

  7. Drugs with anti-oxidant properties can interfere with cell viability measurements by assays that rely on the reducing property of viable cells.

    Science.gov (United States)

    Shenoy, Niraj; Stenson, Mary; Lawson, Joshua; Abeykoon, Jithma; Patnaik, Mrinal; Wu, Xiaosheng; Witzig, Thomas

    2017-02-27

    Cell viability assays such as Cell Titer Blue and Alamar Blue rely on the reducing property of viable cells to reduce the reagent dye to a product which gives a fluorescent signal. The current manufacture-recommended protocols do not take into account the possibility of the reagent substrate being reduced directly to the fluorescent product by drugs with an anti-oxidant property. After suspecting spurious results while determining the cytotoxic potential of a drug of interest (DOI) with known anti-oxidant property against a renal cell cancer (RCC) cell line, we aimed to establish that drugs with anti-oxidant property can indeed cause false-negative results with the current protocols of these assays by direct reduction of the reagent substrate. We also aimed to counter the same with a simple modification added to the protocol. Through our experiments, we conclusively demonstrate that drugs with anti-oxidant properties can indeed interfere with cell viability measurements by assays that rely on the reducing property of viable cells. A simple modification in the protocol, as elaborated in the manuscript, can prevent spurious results with these otherwise convenient assays.Laboratory Investigation advance online publication, 27 February 2017; doi:10.1038/labinvest.2017.18.

  8. A Novel Application for Low Frequency Electrochemical Impedance Spectroscopy as an Online Process Monitoring Tool for Viable Cell Concentrations

    Directory of Open Access Journals (Sweden)

    Christoph Slouka

    2016-11-01

    Full Text Available New approaches in process monitoring during industrial fermentations are not only limited to classical pH, dO2 and offgas analysis, but use different in situ and online sensors based on different physical principles to determine biomass, product quality, lysis and far more. One of the very important approaches is the in situ accessibility of viable cell concentration (VCC. This knowledge provides increased efficiency in monitoring and controlling strategies during cultivations. Electrochemical impedance spectroscopy—EIS—is used to monitor biomass in a fermentation of E. coli BL21(DE3, producing a recombinant protein using a fed batch-based approach. Increases in the double layer capacitance (Cdl, determined at frequencies below 1 kHz, are proportional to the increase of biomass in the batch and fed batch phase, monitored in offline and online modes for different cultivations. A good correlation of Cdl with cell density is found and in order to get an appropriate verification of this method, different state-of-the-art biomass measurements are performed and compared. Since measurements in this frequency range are largely determined by the double layer region between the electrode and media, rather minor interferences with process parameters (aeration, stirring are to be expected. It is shown that impedance spectroscopy at low frequencies is a powerful tool for cultivation monitoring.

  9. Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.

    Directory of Open Access Journals (Sweden)

    Bee Luan Khoo

    Full Text Available Circulating tumor cells (CTCs are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughput, selectivity and specificity of devices or assays used in CTC detection and isolation.Here, we enriched and characterized putative CTCs from blood samples of patients with both advanced stage metastatic breast and lung cancers using a novel multiplexed spiral microfluidic chip. This system detected putative CTCs under high sensitivity (100%, n = 56 (Breast cancer samples: 12-1275 CTCs/ml; Lung cancer samples: 10-1535 CTCs/ml rapidly from clinically relevant blood volumes (7.5 ml under 5 min. Blood samples were completely separated into plasma, CTCs and PBMCs components and each fraction were characterized with immunophenotyping (Pan-cytokeratin/CD45, CD44/CD24, EpCAM, fluorescence in-situ hybridization (FISH (EML4-ALK or targeted somatic mutation analysis. We used an ultra-sensitive mass spectrometry based system to highlight the presence of an EGFR-activating mutation in both isolated CTCs and plasma cell-free DNA (cf-DNA, and demonstrate concordance with the original tumor-biopsy samples.We have clinically validated our multiplexed microfluidic chip for the ultra high-throughput, low-cost and label-free enrichment of CTCs. Retrieved cells were unlabeled and viable, enabling potential propagation and real-time downstream analysis using next generation sequencing (NGS or proteomic analysis.

  10. Quantitation of viable Coxiella burnetii in milk using an integrated cell culture-polymerase chain reaction (ICC-PCR) assay.

    Science.gov (United States)

    Stewart, Diana; Shieh, Y-Carol; Tortorello, Mary; Kukreja, Ankush; Shazer, Arlette; Schlesser, Joseph

    2015-11-01

    The obligate intracellular pathogen Coxiella burnetii has long been considered the most heat resistant pathogen in raw milk, making it the reference pathogen for determining pasteurisation conditions for milk products. New milk formulations and novel non-thermal processes require validation of effectiveness which requires a more practical method for analysis than using the currently used animal model for assessing Coxiella survival. Also, there is an interest in better characterising thermal inactivation of Coxiella in various milk formulations. To avoid the use of the guinea pig model for evaluating Coxiella survival, an Integrated Cell Culture-PCR (ICC-PCR) method was developed for determining Coxiella viability in milk. Vero cell cultures were directly infected from Coxiella-contaminated milk in duplicate 24-well plates. Viability of the Coxiella in milk was shown by a ≥ 0.5 log genome equivalent (ge)/ml increase in the quantity of IS111a gene from the baseline post-infection (day 0) level after 9-11 d propagation. Coxiella in skim, 2%, and whole milk, and half and half successfully infected Vero cells and increased in number by at least 2 logs using a 48-h infection period followed by 9-d propagation time. As few as 125 Coxiella ge/ml in whole milk was shown to infect and propagate at least 2 logs in the optimised ICC-PCR assay, though variable confirmation of propagation was shown for as low as 25 Coxiella ge/ml. Applicability of the ICC-PCR method was further proven in an MPN format to quantitate the number of viable Coxiella remaining in whole milk after 60 °C thermal treatment at 0, 20, 40, 60 and 90 min.

  11. Magnetic nanoparticles to recover cellular organelles and study the time resolved nanoparticle-cell interactome throughout uptake.

    Science.gov (United States)

    Bertoli, Filippo; Davies, Gemma-Louise; Monopoli, Marco P; Moloney, Micheal; Gun'ko, Yurii K; Salvati, Anna; Dawson, Kenneth A

    2014-08-27

    Nanoparticles in contact with cells and living organisms generate quite novel interactions at the interface between the nanoparticle surface and the surrounding biological environment. However, a detailed time resolved molecular level description of the evolving interactions as nanoparticles are internalized and trafficked within the cellular environment is still missing and will certainly be required for the emerging arena of nanoparticle-cell interactions to mature. In this paper promising methodologies to map out the time resolved nanoparticle-cell interactome for nanoparticle uptake are discussed. Thus silica coated magnetite nanoparticles are presented to cells and their magnetic properties used to isolate, in a time resolved manner, the organelles containing the nanoparticles. Characterization of the recovered fractions shows that different cell compartments are isolated at different times, in agreement with imaging results on nanoparticle intracellular location. Subsequently the internalized nanoparticles can be further isolated from the recovered organelles, allowing the study of the most tightly nanoparticle-bound biomolecules, analogous to the 'hard corona' that so far has mostly been characterized in extracellular environments. Preliminary data on the recovered nanoparticles suggest that significant portion of the original corona (derived from the serum in which particles are presented to the cells) is preserved as nanoparticles are trafficked through the cells. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Polyelectrolyte Complex Beads by Novel Two-Step Process for Improved Performance of Viable Whole-Cell Baeyer-Villiger Monoxygenase by Immobilization

    Directory of Open Access Journals (Sweden)

    Tomáš Krajčovič

    2017-11-01

    Full Text Available A novel immobilization matrix for the entrapment of viable whole-cell Baeyer–Villiger monooxygenase was developed. Viable recombinant Escherichia coli cells overexpressing cyclohexanone monooxygenase were entrapped in polyelectrolyte complex beads prepared by a two-step reaction of oppositely-charged polymers including highly defined cellulose sulphate. Immobilized cells exhibited higher operational stability than free cells during 10 repeated cycles of Baeyer–Villiger biooxidations of rac-bicyclo[3.2.0]hept-2-en-6-one to the corresponding lactones (1R,5S-3-oxabicyclo-[3.3.0]oct-6-en-3-one and (1S,5R-2-oxabicyclo-[3.3.0]oct-6-en-3-one. The morphology of polyelectrolyte complex beads was characterised by environmental scanning electron microscopy; the spatial distribution of polymers in the beads and cell viability were examined using confocal laser scanning microscopy, and the texture was characterised by the mechanical resistance measurements.

  13. Enhanced inhibition of murine prostatic carcinoma growth by immunization with or administration of viable human umbilical vein endothelial cells and CRM197

    Directory of Open Access Journals (Sweden)

    Zhang Huiyong

    2011-02-01

    Full Text Available Vaccination with xenogeneic and syngeneic endothelial cells is effective for inhibiting tumor growth. Nontoxic diphtheria toxin (CRM197, as an immunogen or as a specific inhibitor of heparin-binding EGF-like growth factor, has shown promising antitumor activity. Therefore, immunization with or administration of viable human umbilical vein endothelial cells (HUVECs combined with CRM197 could have an enhanced antitumor effect. Six-week-old C57BL/6J male mice were vaccinated with viable HUVECs, 1 x 10(6 viable HUVECs combined with 100 μg CRM197, or 100 μg CRM197 alone by ip injections once a week for 4 consecutive weeks. RM-1 cells (5 x 10(5 were inoculated by sc injection as a preventive procedure. During the therapeutic procedure, 6-week-old male C57BL/6J mice were challenged with 1 x 10(5 RM-1 cells, then injected sc with 1 x 10(6 viable HUVECs, 1 x 10(6 viable HUVECs + 100 μg CRM197, and 100 μg CRM197 alone twice a week for 4 consecutive weeks. Tumor volume and life span were monitored. We also investigated the effects of immunization with HUVECs on the aortic arch wall and on wound healing. Vaccination with or administration of viable HUVECs+CRM197 enhanced the inhibition of RM-1 prostatic carcinoma by 24 and 29%, respectively, and prolonged the life span for 3 and 4 days, respectively, compared with those of only vaccination or administration with viable HUVECs of tumor-bearing C57BL/6J mice. Furthermore, HUVEC immunization caused some damage to the aortic arch wall but did not have remarkable effects on the rate of wound healing; the wounds healed in approximately 13 days. Treatment with CRM197 in combination with viable HUVECs resulted in a marked enhancement of the antitumor effect in the preventive or therapeutic treatment for prostatic carcinoma in vivo, suggesting a novel combination for anti-cancer therapy.

  14. Assessing prognosis and optimizing treatment in patients with postchemotherapy viable nonseminomatous germ-cell tumors (NSGCT): results of the sCR2 international study

    DEFF Research Database (Denmark)

    Fizazi, K.; Oldenburg, J.; Dunant, A.

    2008-01-01

    malignant cells, and a good International Germ Cell Consensus Classification group at presentation. Patients were assigned to one of three risk groups defined in sCR1: no risk factor (good risk), one risk factor (intermediate risk) and two to three risk factors (poor risk group). The 5-year PFS rate was 92...... with surveillance and treatment only at relapse. CONCLUSION: In patients with postchemotherapy viable NSGCT, a complete resection of residual masses should be rigorously pursued. These data validate the sCR1 prognostic index. Given their excellent outcome, patients in the favorable group may not require......BACKGROUND: The purpose of this study was to validate a prognostic index [surgical complete response 1 (sCR1)] in patients with postchemotherapy viable nonseminomatous germ-cell tumors (NSGCT). PATIENTS AND METHODS: Data and specimens from 61 patients with normalized tumor markers...

  15. Cytokine Profiles and Cell Proliferation Responses to Truncated ORF2 Protein in Iranian Patients Recovered from Hepatitis E Infection.

    Science.gov (United States)

    Taherkhani, Reza; Farshadpour, Fatemeh; Makvandi, Manoochehr; Rajabi Memari, Hamid; Samarbafzadeh, Ali Reza; Sharifi, Nasrin; Naeimi, Behrouz; Tajbakhsh, Saeed; Akbarzadeh, Samad

    2015-01-01

    Background. The aim of this study was to evaluate hepatitis E virus (HEV) specific cellular immune responses to truncated ORF2 protein in Iranian patients recovered from HEV infection. Information about HEV-specific immune responses could be useful in finding an effective way for development of HEV vaccine. Methods. A truncated form of HEV ORF2 protein containing amino acids 112-608 was used to stimulate peripheral blood mononuclear cells (PBMCs) separated from HEV-recovered and control groups. Finally, the levels of four cytokines, IFN-γ ELISPOT, and cell proliferative responses following stimulation with the truncated ORF2 protein were assessed in the both groups. Results. The truncated ORF2 protein was able to induce IFN-γ ELISPOT and cell proliferation responses and to produce significant amounts of IFN-γ and IL-12 cytokines, but low amounts of IL-10 and IL-4 cytokines in vitro. These responses were significantly higher in the recovered group compared to the control group. These results indicate the antigenic nature of the truncated ORF2 protein and production of T helper type 1 cytokines. Conclusion. The truncated ORF2 protein can effectively induce significant cellular immune responsesand can be introduced as a potential vaccine candidate. However, further studies are required to evaluate this protein in vivo.

  16. Cytokine Profiles and Cell Proliferation Responses to Truncated ORF2 Protein in Iranian Patients Recovered from Hepatitis E Infection

    Directory of Open Access Journals (Sweden)

    Reza Taherkhani

    2015-01-01

    Full Text Available Background. The aim of this study was to evaluate hepatitis E virus (HEV specific cellular immune responses to truncated ORF2 protein in Iranian patients recovered from HEV infection. Information about HEV-specific immune responses could be useful in finding an effective way for development of HEV vaccine. Methods. A truncated form of HEV ORF2 protein containing amino acids 112-608 was used to stimulate peripheral blood mononuclear cells (PBMCs separated from HEV-recovered and control groups. Finally, the levels of four cytokines, IFN-γ ELISPOT, and cell proliferative responses following stimulation with the truncated ORF2 protein were assessed in the both groups. Results. The truncated ORF2 protein was able to induce IFN-γ ELISPOT and cell proliferation responses and to produce significant amounts of IFN-γ and IL-12 cytokines, but low amounts of IL-10 and IL-4 cytokines in vitro. These responses were significantly higher in the recovered group compared to the control group. These results indicate the antigenic nature of the truncated ORF2 protein and production of T helper type 1 cytokines. Conclusion. The truncated ORF2 protein can effectively induce significant cellular immune responsesand can be introduced as a potential vaccine candidate. However, further studies are required to evaluate this protein in vivo.

  17. Viable Cancer Cells in the Remnant Stomach are a Potential Source of Peritoneal Metastasis after Curative Distal Gastrectomy for Gastric Cancer.

    Science.gov (United States)

    Murata, Satoshi; Yamamoto, Hiroshi; Yamaguchi, Tsuyoshi; Kaida, Sachiko; Ishida, Mitsuaki; Kodama, Hirokazu; Takebayashi, Katsushi; Shimizu, Tomoharu; Miyake, Toru; Tani, Tohru; Kushima, Ryoji; Tani, Masaji

    2016-09-01

    The mechanisms underlying peritoneal metastasis (PM) after curative gastrectomy for gastric cancer (GC) are not well elucidated. This study assessed whether viable cancer cells, including cancer stemlike cells (CSCs), were present in the remnant stomach immediately before gastrointestinal (GI) tract reconstruction because these could be a source of PM after gastrectomy. Saline fluid used for remnant stomach lumen irrigation before GI reconstruction was prospectively collected from 142 consecutive patients undergoing distal gastrectomy for GC and cytologically examined. Proliferative activity (Ki67 staining) and stemness (expression of the CSC surface markers CD44s or CD44v6) were evaluated in detected cancer cells. Viable cancer cells were detected in 33 (23.2 %) of the 142 remnant stomachs. These cells formed clusters and stained positively for Ki67, indicating proliferation. Cancer cells in remnant stomachs and surface cancer cells in primary GCs from 10 (30.3 %) of these 33 cases also stained positively for CD44s or CD44v6. In a multiple logistic regression analysis, advanced cancer (odds ratio [OR], 4.65; 95 % confidence interval [CI], 1.32-16.4; P = 0.017), tumor size of 40 mm or larger (OR, 3.78; 95 % CI, 1.12-12.8; P = 0.033), and histologic differentiation (OR, 3.10; 95 % CI, 1.30-7.40; P = 0.011) were associated independently with the presence of cancer cells in the remnant stomach. Viable, proliferative, and clustered cancer cells, including CSCs, were found in remnant gastric lumens immediately before GI reconstruction, indicating a possible cellular source of PM after curative gastrectomy for GC. Dissemination of gastric contents into the peritoneal cavity should be avoided during GI reconstruction.

  18. Enhancing forward osmosis water recovery from landfill leachate by desalinating brine and recovering ammonia in a microbial desalination cell.

    Science.gov (United States)

    Iskander, Syeed Md; Novak, John T; He, Zhen

    2018-01-31

    In this work, a microbial desalination cell (MDC) was employed to desalinate the FO treated leachate for reduction of both salinity and chemical oxygen demand (COD). The FO recovered 51.5% water from a raw leachate and the recovery increased to 83.5% from the concentrated leachate after desalination in the MDC fed with either acetate or another leachate as an electron source and at a different hydraulic retention time (HRT). Easily-degraded substrate like acetate and a long HRT resulted in a low conductivity desalinated effluent. Ammonia was also recovered in the MDC cathode with a recovery efficiency varying from 11 to 64%, affected by current generation and HRT. Significant COD reduction, as high as 65.4%, was observed in the desalination chamber and attributed to the decrease of both organic and inorganic compounds via diffusion and electricity-driven movement. Copyright © 2018 Elsevier Ltd. All rights reserved.

  19. Designing primers and evaluation of the efficiency of propidium monoazide – Quantitative polymerase chain reaction for counting the viable cells of Lactobacillus gasseri and Lactobacillus salivarius

    Directory of Open Access Journals (Sweden)

    Chieh-Hsien Lai

    2017-07-01

    Full Text Available The purpose of this study is to evaluate the efficiency of using propidium monoazide (PMA real-time quantitative polymerase chain reaction (qPCR to count the viable cells of Lactobacillus gasseri and Lactobacillus salivarius in probiotic products. Based on the internal transcription spacer and 23S rRNA genes, two primer sets specific for these two Lactobacillus species were designed. For a probiotic product, the total deMan Rogosa Sharpe plate count was 8.65±0.69 log CFU/g, while for qPCR, the cell counts of L. gasseri and L. salivarius were 8.39±0.14 log CFU/g and 8.57±0.24 log CFU/g, respectively. Under the same conditions, for its heat-killed product, qPCR counts for L. gasseri and L. salivarius were 6.70±0.16 log cells/g and 7.67±0.20 log cells/g, while PMA-qPCR counts were 5.33±0.18 log cells/g and 5.05±0.23 log cells/g, respectively. For cell dilutions with a viable cell count of 8.5 log CFU/mL for L. gasseri and L. salivarius, after heat killing, the PMA-qPCR count for both Lactobacillus species was near 5.5 log cells/mL. When the PMA-qPCR counts of these cell dilutions were compared before and after heat killing, although some DNA might be lost during the heat killing, significant qPCR signals from dead cells, i.e., about 4–5 log cells/mL, could not be reduced by PMA treatment. Increasing PMA concentrations from 100 μM to 200 μM or light exposure time from 5 minutes to 15 minutes had no or, if any, only minor effect on the reduction of qPCR signals from their dead cells. Thus, to differentiate viable lactic acid bacterial cells from dead cells using the PMA-qPCR method, the efficiency of PMA to reduce the qPCR signals from dead cells should be notable.

  20. Designing primers and evaluation of the efficiency of propidium monoazide - Quantitative polymerase chain reaction for counting the viable cells of Lactobacillus gasseri and Lactobacillus salivarius.

    Science.gov (United States)

    Lai, Chieh-Hsien; Wu, Sih-Rong; Pang, Jen-Chieh; Ramireddy, Latha; Chiang, Yu-Cheng; Lin, Chien-Ku; Tsen, Hau-Yang

    2017-07-01

    The purpose of this study is to evaluate the efficiency of using propidium monoazide (PMA) real-time quantitative polymerase chain reaction (qPCR) to count the viable cells of Lactobacillus gasseri and Lactobacillus salivarius in probiotic products. Based on the internal transcription spacer and 23S rRNA genes, two primer sets specific for these two Lactobacillus species were designed. For a probiotic product, the total deMan Rogosa Sharpe plate count was 8.65±0.69 log CFU/g, while for qPCR, the cell counts of L. gasseri and L. salivarius were 8.39±0.14 log CFU/g and 8.57±0.24 log CFU/g, respectively. Under the same conditions, for its heat-killed product, qPCR counts for L. gasseri and L. salivarius were 6.70±0.16 log cells/g and 7.67±0.20 log cells/g, while PMA-qPCR counts were 5.33±0.18 log cells/g and 5.05±0.23 log cells/g, respectively. For cell dilutions with a viable cell count of 8.5 log CFU/mL for L. gasseri and L. salivarius, after heat killing, the PMA-qPCR count for both Lactobacillus species was near 5.5 log cells/mL. When the PMA-qPCR counts of these cell dilutions were compared before and after heat killing, although some DNA might be lost during the heat killing, significant qPCR signals from dead cells, i.e., about 4-5 log cells/mL, could not be reduced by PMA treatment. Increasing PMA concentrations from 100 μM to 200 μM or light exposure time from 5 minutes to 15 minutes had no or, if any, only minor effect on the reduction of qPCR signals from their dead cells. Thus, to differentiate viable lactic acid bacterial cells from dead cells using the PMA-qPCR method, the efficiency of PMA to reduce the qPCR signals from dead cells should be notable. Copyright © 2016. Published by Elsevier B.V.

  1. Not single but periodic injections of synovial mesenchymal stem cells maintain viable cells in knees and inhibit osteoarthritis progression in rats.

    Science.gov (United States)

    Ozeki, N; Muneta, T; Koga, H; Nakagawa, Y; Mizuno, M; Tsuji, K; Mabuchi, Y; Akazawa, C; Kobayashi, E; Matsumoto, K; Futamura, K; Saito, T; Sekiya, I

    2016-06-01

    We investigated the effects of single or repetitive intra-articular injections of synovial mesenchymal stem cells (MSCs) on a rat osteoarthritis (OA) model, and elucidated the behaviors and underlying mechanisms of the stem cells after the injection. One week after the transection of the anterior cruciate ligament (ACL) of wild type Lewis rats, one million synovial MSCs were injected into the knee joint every week. Cartilage degeneration was evaluated with safranin-o staining after the first injection. To analyze cell kinetics or MSC properties, luciferase, LacZ, and GFP expressing synovial MSCs were used. To confirm the role of MSCs, species-specific microarray and PCR analyses were performed using human synovial MSCs. Histological analysis for femoral and tibial cartilage showed that a single injection was ineffective but weekly injections had significant chondroprotective effects for 12 weeks. Histological and flow-cytometric analyses of LacZ and GFP expressing synovial MSCs revealed that injected MSCs migrated mainly into the synovium and most of them retained their undifferentiated MSC properties though the migrated cells rapidly decreased. In vivo imaging analysis revealed that MSCs maintained in knees while weekly injection. Species-specific microarray and PCR analyses showed that the human mRNAs on day 1 for 21 genes increased over 50-fold, and increased the expressions of PRG-4, BMP-2, and BMP-6 genes encoding chondroprotective proteins, and TSG-6 encoding an anti-inflammatory one. Not single but periodic injections of synovial MSCs maintained viable cells without losing their MSC properties in knees and inhibited osteoarthritis (OA) progression by secretion of trophic factors. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  2. Macrophage conditioned medium induced cellular network formation in MCF-7 cells through enhanced tunneling nanotube formation and tunneling nanotube mediated release of viable cytoplasmic fragments

    Energy Technology Data Exchange (ETDEWEB)

    Patheja, Pooja, E-mail: pooja.patheja8@gmail.com [Laser Biomedical Applications Section, Raja Ramanna Centre for Advanced Technology, Indore 452013, Madhya Pradesh (India); Homi Bhabha National Institute, Training School Complex, Anushaktinagar, Mumbai 400094, Maharashtra (India); Sahu, Khageswar [Laser Biomedical Applications Section, Raja Ramanna Centre for Advanced Technology, Indore 452013, Madhya Pradesh (India)

    2017-06-15

    Infiltrating macrophages in tumor microenvironment, through their secreted cytokines and growth factors, regulate several processes of cancer progression such as cancer cell survival, proliferation, invasion, metastasis and angiogenesis. Recently, intercellular cytoplasmic bridges between cancer cells referred as tunneling nanotubes (TNTs) have been recognized as novel mode of intercellular communication between cancer cells. In this study, we investigated the effect of inflammatory mediators present in conditioned medium derived from macrophages on the formation of TNTs in breast adenocarcinoma cells MCF-7. Results show that treatment with macrophage conditioned medium (MφCM) not only enhanced TNT formation between cells but also stimulated the release of independently migrating viable cytoplasmic fragments, referred to as microplasts, from MCF-7 cells. Time lapse microscopy revealed that microplasts were released from parent cancer cells in extracellular space through formation of TNT-like structures. Mitochondria, vesicles and cytoplasm could be transferred from parent cell body to microplasts through connecting TNTs. The microplasts could also be resorbed into the parent cell body by retraction of the connecting TNTs. Microplast formation inhibited in presence cell migration inhibitor, cytochalasin-B. Notably by utilizing migratory machinery within microplasts, distantly located MCF-7 cells formed several TNT based intercellular connections, leading to formation of physically connected network of cells. Together, these results demonstrate novel role of TNTs in microplast formation, novel modes of TNT formation mediated by microplasts and stimulatory effect of MφCM on cellular network formation in MCF-7 cells mediated through enhanced TNT and microplast formation.

  3. Invariant natural killer T cells are depleted in renal impairment and recover after kidney transplantation.

    Science.gov (United States)

    Peukert, Konrad; Wingender, Gerhard; Patecki, Margret; Wagner, Stephan; Schmitt, Roland; Ge, Shuwang; Schwarz, Anke; Kronenberg, Mitchell; Haller, Hermann; von Vietinghoff, Sibylle

    2014-05-01

    Altered immune function in patients with renal failure results in both susceptibility to infection and increased inflammatory response. Invariant natural killer T (iNKT) cells are a conserved, immunoregulatory T lymphocyte subset that responds to lipid antigens with near-immediate cytokine production and cytotoxicity. iNKT cells are required for the antibacterial host response. Whether renal failure and renal replacement therapy alter iNKT cell abundance or phenotype has not been investigated. iNKT cells were studied by flow cytometry in the peripheral blood of patients with acute renal failure, chronic haemo- and peritoneal dialysis (PD), chronic kidney disease and after renal transplantation. A very marked reduction in iNKT lymphocytes was found in acute renal failure before the first haemodialysis (HD) session. iNKT cells were depleted in end-stage renal disease patients receiving either HD or PD. iNKT cell depletion was accentuated after an HD session. Lesser degrees were observed in patients with non-dialysis-dependent chronic kidney disease. CD56 and CD161 NK cell marker expression was decreased in renal impairment. CD56(+) and CD161(+) iNKT cells produced more interferon-γ than negative cells of the same donor. Within the first year after kidney transplantation, the decrease in iNKT cells and their NK cell markers was reverted. We describe for the first time that iNKT lymphocytes are reduced in end-stage renal disease and further depleted by HD. iNKT cells are important for early host response including activation of other immune cells and their depletion may contribute to immune dysfunction in renal disease.

  4. DEMONSTRATION OF FUEL CELLS TO RECOVER ENERGY FROM LANDFILL GAS - PHASE I FINAL REPORT: CONCEPTUAL STUDY

    Science.gov (United States)

    The report discusses results of a conceptual design, cost, and evaluation study of energy recovery from landfill gas using a commercial phosphoric acid fuel cell power plant. The conceptual design of the fuel cell energy recovery system is described, and its economic and environm...

  5. Increased cell proliferation and mucocyte density in the sea anemone Aiptasia pallida recovering from bleaching.

    Science.gov (United States)

    Fransolet, David; Roberty, Stéphane; Herman, Anne-Catherine; Tonk, Linda; Hoegh-Guldberg, Ove; Plumier, Jean-Christophe

    2013-01-01

    Recovery of coral after bleaching episodes is a critical period for the health of the reef ecosystem. While events such as symbiont (genus Symbiodinium) shifting/shuffling or tissue apoptosis have been demonstrated to occur following bleaching, little is known concerning tissue recovery or cell proliferation. Here, we studied the sea anemone Aiptasia pallida exposed to a transient elevation of water temperature combined with high illumination (33°C and 1900 µmol photons x m(-2) x s(-1) for 30 h). Following such treatment bleached anemones showed a significant reduction of their Symbiodinium density. Cell proliferation in the ectodermis and gastrodermis was determined by assessing the densities of cells labeled with a thymidine analogue (EdU). Cell proliferation significantly increased during the first day following stress in both tissue types. This increased cell proliferation returned to pre-stress values after one week. Although cell proliferation was higher in the ectodermis in absence of stress, it was relatively more pronounced in the gastrodermis of stressed anemones. In addition, the ratio of ectodermal mucocytes significantly increased three weeks after induced stress. These results suggest that thermal/photic stress coupled with the loss of the symbionts is able to enhance cell proliferation in both gastrodermis and ectodermis of cnidarians. While new cells formed in the gastrodermis are likely to host new Symbiodinium, the fate of new cells in the ectodermis was only partially revealed. Some new ectodermal cells may, in part, contribute to the increased number of mucocytes which could eventually help strengthen the heterotrophic state until restoration of the symbiosis.

  6. Increased cell proliferation and mucocyte density in the sea anemone Aiptasia pallida recovering from bleaching.

    Directory of Open Access Journals (Sweden)

    David Fransolet

    Full Text Available Recovery of coral after bleaching episodes is a critical period for the health of the reef ecosystem. While events such as symbiont (genus Symbiodinium shifting/shuffling or tissue apoptosis have been demonstrated to occur following bleaching, little is known concerning tissue recovery or cell proliferation. Here, we studied the sea anemone Aiptasia pallida exposed to a transient elevation of water temperature combined with high illumination (33°C and 1900 µmol photons x m(-2 x s(-1 for 30 h. Following such treatment bleached anemones showed a significant reduction of their Symbiodinium density. Cell proliferation in the ectodermis and gastrodermis was determined by assessing the densities of cells labeled with a thymidine analogue (EdU. Cell proliferation significantly increased during the first day following stress in both tissue types. This increased cell proliferation returned to pre-stress values after one week. Although cell proliferation was higher in the ectodermis in absence of stress, it was relatively more pronounced in the gastrodermis of stressed anemones. In addition, the ratio of ectodermal mucocytes significantly increased three weeks after induced stress. These results suggest that thermal/photic stress coupled with the loss of the symbionts is able to enhance cell proliferation in both gastrodermis and ectodermis of cnidarians. While new cells formed in the gastrodermis are likely to host new Symbiodinium, the fate of new cells in the ectodermis was only partially revealed. Some new ectodermal cells may, in part, contribute to the increased number of mucocytes which could eventually help strengthen the heterotrophic state until restoration of the symbiosis.

  7. Bacillus subtilis mutants with knockouts of the genes encoding ribonucleases RNase Y and RNase J1 are viable, with major defects in cell morphology, sporulation, and competence.

    Science.gov (United States)

    Figaro, Sabine; Durand, Sylvain; Gilet, Laetitia; Cayet, Nadège; Sachse, Martin; Condon, Ciarán

    2013-05-01

    The genes encoding the ribonucleases RNase J1 and RNase Y have long been considered essential for Bacillus subtilis cell viability, even before there was concrete knowledge of their function as two of the most important enzymes for RNA turnover in this organism. Here we show that this characterization is incorrect and that ΔrnjA and Δrny mutants are both viable. As expected, both strains grow relatively slowly, with doubling times in the hour range in rich medium. Knockout mutants have major defects in their sporulation and competence development programs. Both mutants are hypersensitive to a wide range of antibiotics and have dramatic alterations to their cell morphologies, suggestive of cell envelope defects. Indeed, RNase Y mutants are significantly smaller in diameter than wild-type strains and have a very disordered peptidoglycan layer. Strains lacking RNase J1 form long filaments in tight spirals, reminiscent of mutants of the actin-like proteins (Mre) involved in cell shape determination. Finally, we combined the rnjA and rny mutations with mutations in other components of the degradation machinery and show that many of these strains are also viable. The implications for the two known RNA degradation pathways of B. subtilis are discussed.

  8. Cryo-ablation improves anti-tumor immunity through recovering tumor educated dendritic cells in tumor-draining lymph nodes.

    Science.gov (United States)

    He, Xiao-Zheng; Wang, Qi-Fu; Han, Shuai; Wang, Hui-Qing; Ye, Yong-Yi; Zhu, Zhi-Yuan; Zhang, Shi-Zhong

    2015-01-01

    In addition to minimally invasive destruction of tumors, cryo-ablation of tumors to some extent modulated anti-tumor immunity. Cryo-ablated tumors in glioma mice models induced anti-tumor cellular immunologic response which increases the percentage of CD3(+) and CD4(+)T cells in blood as well as natural killer cells. As a crucial role in triggering anti-tumor immunity, dendritic cells (DCs) were educated by tumors to adopt a tolerance phenotype which helps the tumor escape from immune monitoring. This study aims to study whether cryo-ablation could influence the tolerogenic DCs, and influence anti-tumor immunity in tumor-draining lymph nodes (TDLNs). Using the GL261 subcutaneous glioma mouse model, we created a tumor bearing group, cryo-ablation group, and surgery group. We analyzed alteration in phenotype and function of tolerogenic DCs, and evaluated the factors of anti-tumor immunity inhibition. DCs in TDLNs in GL261 subcutaneous glioma mouse model expressed tolerogenic phenotype. In contrast to surgery, cryo-ablation improved the quantity and quality of these tolerogenic DCs. Moreover, the DCs decreased the expression of intracellular interleukin-10 (IL-10) and extra-cellular IL-10. In vitro, DCs from the cryo-ablation group recovered their specific function and induced potent anti-tumor immunity through triggering T cells. In vivo, cryo-ablation showed weak anti-tumor immunity, only inhibiting the growth of rechallenged tumors. But many IL-10-low DCs, rather than IL-10-high DCs, infiltrated the tumors. More importantly, Tregs inhibited the performance of these DCs; and depletion of Tregs greatly improved anti-tumor immunity in vivo. Cryo-ablation could recover function of tumor induced tolerogenic DCs in vitro; and depletion of Tregs could improve this anti-tumor effect in vivo. The Tregs/CD4(+)T and Tregs/CD25(+)T cells in TDLNs inhibit DCs' activity and function.

  9. Effects of the oral administration of viable and heat-killed Streptococcus bovis HC5 cells to pre-sensitized BALB/c mice.

    Directory of Open Access Journals (Sweden)

    Aline D Paiva

    Full Text Available Antimicrobial peptides have been suggested as an alternative to classical antibiotics in livestock production and bacteriocin-producing bacteria could be added to animal feeds to deliver bacteriocins in the gastrointestinal (GI tract of ruminant and monogastric animals. In this study, viable (V and heat-killed (HK Streptococcus bovis HC5 cells were orally administered to pre-sensitized mice in order to assess the effects of a bacteriocin-producing bacteria on histological parameters and the immune response of the GI tract of monogastric animals. The administration of V and HK S. bovis HC5 cells during 58 days to BALB/c mice did not affect weight gain, but an increase in gut permeability was detected in animals receiving the HK cells. Viable and heat killed cells caused similar morphological alterations in the GI tract of the animals, but the most prominent effects were detected in the small intestine. The oral administration of S. bovis HC5 also influenced cytokine production in the small intestine, and the immune-mediated activity differed between V and HK cells. The relative expression of IL-12 and INF-γ was significantly higher in the small intestine of mice treated with V cells, while an increase in IL-5, IL-13 and TNF-α expression was only detected in mice treated with HK cells. Considering that even under a condition of severe challenge (pre-sensitization followed by daily exposure to the same bacterial immunogen the general health of the animals was maintained, it appears that oral administration of S. bovis HC5 cells could be a useful route to deliver bacteriocin in the GI tract of livestock animals.

  10. A non-aggressive, highly efficient, enzymatic method for dissociation of human brain-tumors and brain-tissues to viable single-cells.

    Science.gov (United States)

    Volovitz, Ilan; Shapira, Netanel; Ezer, Haim; Gafni, Aviv; Lustgarten, Merav; Alter, Tal; Ben-Horin, Idan; Barzilai, Ori; Shahar, Tal; Kanner, Andrew; Fried, Itzhak; Veshchev, Igor; Grossman, Rachel; Ram, Zvi

    2016-06-01

    Conducting research on the molecular biology, immunology, and physiology of brain tumors (BTs) and primary brain tissues requires the use of viably dissociated single cells. Inadequate methods for tissue dissociation generate considerable loss in the quantity of single cells produced and in the produced cells' viability. Improper dissociation may also demote the quality of data attained in functional and molecular assays due to the presence of large quantities cellular debris containing immune-activatory danger associated molecular patterns, and due to the increased quantities of degraded proteins and RNA. Over 40 resected BTs and non-tumorous brain tissue samples were dissociated into single cells by mechanical dissociation or by mechanical and enzymatic dissociation. The quality of dissociation was compared for all frequently used dissociation enzymes (collagenase, DNase, hyaluronidase, papain, dispase) and for neutral protease (NP) from Clostridium histolyticum. Single-cell-dissociated cell mixtures were evaluated for cellular viability and for the cell-mixture dissociation quality. Dissociation quality was graded by the quantity of subcellular debris, non-dissociated cell clumps, and DNA released from dead cells. Of all enzymes or enzyme combinations examined, NP (an enzyme previously not evaluated on brain tissues) produced dissociated cell mixtures with the highest mean cellular viability: 93 % in gliomas, 85 % in brain metastases, and 89 % in non-tumorous brain tissue. NP also produced cell mixtures with significantly less cellular debris than other enzymes tested. Dissociation using NP was non-aggressive over time-no changes in cell viability or dissociation quality were found when comparing 2-h dissociation at 37 °C to overnight dissociation at ambient temperature. The use of NP allows for the most effective dissociation of viable single cells from human BTs or brain tissue. Its non-aggressive dissociative capacity may enable ambient

  11. Prognostic impact of the number of viable circulating cells with high telomerase activity in gastric cancer patients: a prospective study.

    Science.gov (United States)

    Ito, Hiroaki; Inoue, Haruhiro; Kimura, Satoshi; Ohmori, Tohru; Ishikawa, Fumihiro; Gohda, Keigo; Sato, Jun

    2014-07-01

    The identification of circulating tumor cells (CTCs) in peripheral blood is a useful approach to estimate prognosis, monitor disease progression and measure treatment effects in several types of malignancies. We have previously used OBP-401, a telomerase-specific, replication-selective, oncolytic adenoviral agent carrying the green fluorescent protein (GFP) gene. GFP-positive cells (GFP+ cells) were counted under a fluorescence microscope. Our results showed that the number of at least 7.735 µm in diameter GFP+ cells (L-GFP+ cells) in the peripheral blood was a significant marker of prognosis in gastric cancer patients. However, tumor cells undergoing epithelial-mesenchymal transition (EMT) have been reported to be smaller in size than cells without EMT features; thus, CTCs undergoing EMT may escape detection with this technique. Therefore, in this study, we analyzed the relationship between patient outcome and the number of GFP+ cells of any size. We obtained peripheral blood samples from 65 patients with gastric cancer. After infection of OBP-401, GFP+ cells were counted and measured. The relationship between the number of GFP+ cells and surgical outcome was analyzed. The median follow-up period of the surviving patients was 36 months. A significant difference in overall survival was found between patients with 0-5 and patients with ≥6 L-GFP+ cells. No clear relationship was established between the number of small-sized GFP+ cells and patient prognosis. The number of L-GFP+ cells was significantly related to overall survival in patients with gastric cancer. The detection of L-GFP+ cells using OBP-401 may be a useful prognostic marker in gastric cancer.

  12. Human mesenchymal stromal cell lysates as a novel strategy to recover liver function.

    Science.gov (United States)

    Herencia, Carmen; Almadén, Yolanda; Martínez-Moreno, Julio M; Espejo, Isabel; Herrera, Concha; Pérez-Sánchez, Carlos; Guerrero, Fátima; Ciria, Rubén; Briceño, Francisco Javier; Ferrín, Gustavo; de la Mata, Manuel; Muñoz-Castañeda, Juan R

    2015-01-01

    It is unknown if the beneficial effects of mesenchymal stromal cells (MSC) transplantation into the liver are dependent on their anchorage and differentiation into hepatocytes or rather the result of the release of stem cell intracellular content with hepatoprotector properties. The effects of intact MSC transplantation were compared with the infusion of MSC lysates in an experimental rat model of acute liver failure. A more powerful hepatoprotective and antiapoptotic effect was obtained after infusion of MSC lysates than intact MSC. Changes in IL-6 levels and miRNAs might explain the beneficial effects of MSC lysates. Infusion of MSC lysates show a better hepatoprotective effect than the transplantation of intact MSC.

  13. Recovering Citizenship.

    Science.gov (United States)

    Rowe, Michael; Davidson, Larry

    2016-01-01

    Recovery as a process through which people reclaim their lives even while continuing to experience symptoms of mental illness has at times been misunderstood or misinterpreted, including being used as an excuse for cutting needed services and supports in the guise of allowing people to pursue their own personal "recovery journeys." In delivering a message of hope to individuals and fostering initiatives to counter the debilitating impact of clinical pessimism, recovery literature has also, at times, paid less attention to the material, social, cultural, political and economic contexts in which people pursue recovery. The importance of these contexts are implied in a 1961 American Report on Mental Illness and Health that argued that people with mental illnesses should be able to live their lives "in the normal manner" in their home communities. We argue for a disability model that provides people with necessary supports and services to make individual recovery possible for persons with prolonged mental illnesses. Another model that has emerged over the past decade to counterbalance this overweening emphasis on the lone individual pursuing his or her recovery journey is citizenship. We discuss this concept and our research on it since the late 1990s. We then argue for "recovering citizenship" as a concept and metaphor to capture the individual recovery process within the context and goal of a life in the community that the citizenship framework supports.

  14. Binding of CLL subset 4 B-cell receptor immunoglobulins to viable human memory B lymphocytes requires a distinctive IGKV somatic mutation.

    Science.gov (United States)

    Catera, Rosa; Liu, Yun; Gao, Chao; Yan, Xiao-Jie; Magli, Amanda; Allen, Steven L; Kolitz, Jonathan E; Rai, Kanti R; Chu, Charles C; Feizi, Ten; Stamatopoulos, Kostas; Chiorazzi, Nicholas

    2017-01-12

    Amino acid replacement mutations in certain CLL stereotyped B-cell receptor (BCR) immunoglobulins (IGs) at defined positions within antigen-binding sites strongly imply antigen selection. Prime examples of this are CLL subset 4 BCR IGs using IGHV4-34/IGHD5-18/IGHJ6 and IGKV2-30/IGKJ2 rearrangements. Conspicuously and unlike most CLL IGs, subset 4 IGs do not bind apoptotic cells. By testing the (auto)antigenic reactivities of subset 4 IGs toward viable lymphoid-lineage cells and specific autoantigens typically bound by IGHV4-34+ IGs, we found IGs from both subset 4 and non-subset 4 IGHV4-34-expressing CLL cases bind naïve B cells. However, only subset 4 IGs react with memory B cells. Furthermore, subset 4 IGs do not bind DNA nor i or I carbohydrate antigens, common targets of IGHV4-34-utilizing antibodies in systemic lupus erythematosus and cold agglutinin disease, respectively. Notably, we found that subset 4 IG binding to memory B lymphocytes depends on an aspartic acid at position 66 of FR3 in the rearranged IGKV2-30 gene; this amino acid residue is acquired by somatic mutation. Our findings illustrate the importance of positive and negative selection criteria for structural elements in CLL IGs and suggest that autoantigens driving normal B cells to become subset 4 CLL cells differ from those driving IGHV4-34+ B cells in other diseases.

  15. Methods to recover the narrow Dicke sub-Doppler feature in evacuated wall-coated cells without restrictions on cell size

    Science.gov (United States)

    Robinson, H. G.

    1984-01-01

    The hyperfine resonance observed in evacuated wall-coated cells with dimensions lambda/2 (lambda is the hyperfine resonance wavelength) consists of a narrow Dicke sub-Doppler linewidth feature, the spike, superimposed on a broad pedestal. The hydrogen maser provides a classic example of this lineshape. As cell size is increased, an effect unique to evacuated wall-coated cells occurs. Certain combinations of microwave field distribution and cell size result in a lineshape having a pedestal with a small spike feature or only the broad pedestal with no spike. Such conditions are not appropriate for atomic frequency standard applications. The cause of the evacuated wall-coated cell lineshape is reviewed and methods to recover the narrow spike feature without restrictions on cell size is discussed. One example is a cell with dimensions having equal volumes of exposure to opposite phases of the microwave magnetic field.

  16. Fluorescence Quenching Property of C-Phycocyanin from Spirulina platensis and its Binding Efficacy with Viable Cell Components.

    Science.gov (United States)

    Paswan, Meenakshi B; Chudasama, Meghna M; Mitra, Madhusree; Bhayani, Khushbu; George, Basil; Chatterjee, Shruti; Mishra, Sandhya

    2016-03-01

    Phycocyanin is a natural brilliant blue colored, fluorescent protein, which is commonly present in cyanobacteria. In this study, C-phycocyanin was extracted and purified from Spirulina platensis, which are multicellular and filamentous cyanobacteria of greater importance because of its various biological and pharmacological potential. It was analyzed for its binding affinity towards blood cells, algal cells, genomic DNA of microalgae, and bacteria at different temperature and incubation time. It showed good binding affinity with these components even at low concentration of 2.5 μM. The purpose of this study was to evaluate the applicability of C-phycocyanin as a green fluorescent dye substituting carcinogenic chemical dyes.

  17. Managing Viable Knowledge

    NARCIS (Netherlands)

    Achterbergh, J.M.I.M.; Vriens, D.J.

    2002-01-01

    In this paper, Beer's Viable System Model (VSM) is applied to knowledge management. Based on the VSM, domains of knowledge are identified that an organization should possess to maintain its viability. The logic of the VSM is also used to support the diagnosis, design and implementation of the

  18. Determination of Viable Salmonella Typhimurium Cells in Heat Treated Milk By PMA/Real-Time PCR Method

    Directory of Open Access Journals (Sweden)

    Zülal Kesmen

    2017-06-01

    Full Text Available Applying different technological processes during the production of food has a lethal effect on the bacteria but DNA of these bacterial strains may cause false positive results when detected by real time PCR technique because they preserve their existence for a certain period of time. To overcome this shortcoming of the real time PCR technique, a new method has been developed in recent years, based on the removal of dead cell DNA from the medium by treatment with Propodium Monoazide (PMA before DNA extraction. In this study, real-time PCR method was combined with PMA application for the detection of live cells of Salmonella Typhimurium in heat treated milk samples. For this purpose, milk samples inoculated with S. Tyhimurium were heat treated at different temperatures (60, 65, 70 and 75°C and times (15, 60, 300, 900 sec and number of live bacteria was determined comparatively by direct real-time PCR, PMA/real-time PCR and conventional cultural method. As a result, unlike the direct real time PCR technique, PMA/real-time PCR method prevents to a certain extent of false positive results from dead cells at all tested temperatures and times but higher results were obtained from PMA/real-time PCR method when compared to conventional cultural results. Therefore, further studies should be carried out to optimize the conditions of the PMA application in order to eliminate the high positive results detected by the PMA / real-time PCR method

  19. Advances toward regenerative medicine in the central nervous system: challenges in making stem cell therapy a viable clinical strategy.

    Science.gov (United States)

    Stoll, Elizabeth A

    2014-01-01

    Over recent years, there has been a great deal of interest in the prospects of stem cell-based therapies for the treatment of nervous system disorders. The eagerness of scientists, clinicians, and spin-out companies to develop new therapies led to premature clinical trials in human patients, and now the initial excitement has largely turned to skepticism. Rather than embracing a defeatist attitude or pressing blindly ahead, I argue it is time to evaluate the challenges encountered by regenerative medicine in the central nervous system and the progress that is being made to solve these problems. In the twenty years since the adult brain was discovered to have an endogenous regenerative capacity, much basic research has been done to elucidate mechanisms controlling proliferation and cellular identity; how stem cells may be directed into neuronal lineages; genetic, pharmacological, and behavioral interventions that modulate neurogenic activity; and the exact nature of limitations to regeneration in the adult, aged, diseased and injured CNS. These findings should prove valuable in designing realistic clinical strategies to improve the prospects of stem cell-based therapies. In this review, I discuss how basic research continues to play a critical role in identifying both barriers and potential routes to regenerative therapy in the CNS.

  20. Induction of Viable but Nonculturable Salmonella in Exponentially Grown Cells by Exposure to a Low-Humidity Environment and Their Resuscitation by Catalase.

    Science.gov (United States)

    Morishige, Yuta; Koike, Atsushi; Tamura-Ueyama, Ai; Amano, Fumio

    2017-02-01

    Salmonella is a major cause of foodborne disease that sometimes occurs in massive outbreaks around the world. This pathogen is tolerant of low-humidity conditions. We previously described a method for induction of viable but nonculturable (VBNC) Salmonella enterica serovar Enteritidis by treatment with hydrogen peroxide (H2O2) and subsequent resuscitation with 0.3 mM sodium pyruvate. Here, we report a new method for the induction of the VBNC state in Salmonella Enteritidis cells, one involving dehydration. Exposure of Salmonella Enteritidis cells to dehydration stress under poor nutritional conditions (0.9% [wt/vol] NaCl) and 10 to 20% relative humidity at room temperature decreased the presence of culturable population to 0.0067%, but respiratory and glucose uptake active populations were maintained at 0.46 and 1.12%, respectively, meaning that approximately 1% may have entered the VBNC state. Furthermore, these VBNC cells could be resuscitated to acquire culturability by incubation with catalase in M9 minimal medium without glucose in a manner dependent on the dose of catalase but not sodium pyruvate. These results suggest that a low-humidity environment could cause Salmonella Enteritidis cells to enter the VBNC state and the cells could then be resuscitated for growth by treatment with catalase, suggesting a potential risk of Salmonella Enteritidis to survive in low water activity foods in the VBNC state and to start regrowth for foodborne illness.

  1. Freezing Nitrogen Ethanol Composite May be a Viable Approach for Cryotherapy of Human Giant Cell Tumor of Bone.

    Science.gov (United States)

    Wu, Po-Kuei; Chen, Cheng-Fong; Wang, Jir-You; Chen, Paul Chih-Hsueh; Chang, Ming-Chau; Hung, Shih-Chieh; Chen, Wei-Ming

    2017-06-01

    Liquid nitrogen has been used as adjuvant cryotherapy for treating giant cell tumor (GCT) of bone. However, the liquid phase and ultrafreezing (-196° C) properties increase the risk of damage to the adjacent tissues and may lead to perioperative complications. A novel semisolid cryogen, freezing nitrogen ethanol composite, might mitigate these shortcomings because of less-extreme freezing. We therefore wished to evaluate freezing nitrogen ethanol composite as a coolant to determine its properties in tumor cryoablation. (1) Is freezing nitrogen ethanol composite-mediated freezing effective for tumor cryoablation in an ex vivo model, and if yes, is apoptosis involved in the tumor-killing mechanism? (2) Does freezing nitrogen ethanol composite treatment block neovascularization and neoplastic progression of the grafted GCTs and is it comparable to that of liquid nitrogen in an in vivo chicken model? (3) Can use of freezing nitrogen ethanol composite as an adjuvant to curettage result in successful short-term treatment, defined as absence of GCT recurrence at a minimum of 1 year in a small proof-of-concept clinical series? The cryogenic effect on bone tissue mediated by freezing nitrogen ethanol composite and liquid nitrogen was verified by thermal measurement in a time-course manner. Cryoablation on human GCT tissue was examined ex vivo for effect on morphologic features (cell shrinkage) and DNA fragmentation (apoptosis). The presumed mechanism was investigated by molecular analysis of apoptosis regulatory proteins including caspases 3, 8, and 9 and Bax/Bcl-2. Chicken chorioallantoic membrane was used as an in vivo model to evaluate the effects of freezing nitrogen ethanol composite and liquid nitrogen treatment on GCT-derived neovascularization and tumor neoplasm. A small group of patients with GCT of bone was treated by curettage and adjuvant freezing nitrogen ethanol composite cryotherapy in a proof-of-concept study. Tumor recurrence and perioperative

  2. Quality of raw cow milk in Republic of Macedonia determined through the testing of somatic cell count and total viable count

    Directory of Open Access Journals (Sweden)

    Angelovski Ljupco

    2008-11-01

    Full Text Available Somatic cells count and total viable count are criteria used to estimate the compliance of raw cow milk with the Book of rules for demands for safety and hygiene and procedures for official controls of milk and milk products, Official Gazette of RM 157/2007. According to the given demands, raw milk operators are obliged to conduct all procedures and to guarantee that milk is in compliance with the criteria laid down in Book of rules. At the same time, Republic of Macedonia have to fulfill EU criteria laid down in Directive 92/46 (Council directive 92/46/EEC laying down the health rules for the production and placing on the market of raw milk, heat-treated milk and milkbased products for quality of raw milk as part of implementation of community legislation and milk production. The independent laboratory for milk quality control at FVM-Skopje, in frame of its activities in the period February- August 2008 has conducted a study for obtaining preliminary results for the situation with raw milk quality produced in R. of Macedonia for somatic cells counts and total viable count. In the study we analyzed 2065 samples for TVC and 1625 samples for SCC of raw milk samples produced in different parts of the country. From the tested samples only 41,8% fulfill criteria for SCC and 41,45% criteria for TVC lay down in Book of rules for 2008. Assessment of the results in light of Council Directive it is obvious that only 42,7% of the samples for SCC and 10,7% for TVC fulfill the criteria of Council Directive having in mind different requirements vs. Book of rules.

  3. Biologic properties of viable deletion mutants of simian virus 40 (SV40) rescued from the cells of an SV40-induced hamster lymphocytic leukemia.

    Science.gov (United States)

    Diamandopoulos, G T; Carmichael, G

    1983-12-01

    A lymphocytic leukemia induced by the oncogenic DNA simian virus 40 (SV40) in an inbred LSH/SsLak Syrian golden hamster was evoked to produce infectious SV40 by fusion of the leukemia cells with grivet monkey kidney (GMK) cells and by exposure of the leukemia cells to the chemical inducers mitomycin C and cycloheximide. Plaque-purified viable substrains of the rescued SV40 when studied by restriction endonuclease digestion of viral DNA were found to contain small deletions within the Hind III restriction fragment C. These deletions lay near the viral origin of DNA replication. Ten plaque-purified substrains of the rescued virus identified by immunofluorescence as being SV40 were found, when compared to the wild-type SV40, to replicate slowly and to form small plaques. Although these substrains transformed NIH/3T3 cells as efficiently as the wild-type SV40 in tissue culture, they were generally less oncogenic in vivo--7 of the 10 failed to induce tumors. The 3 oncogenic SV40-rescued substrains were not found to exhibit "lymphocytotropism," i.e., the capacity to infect and neoplastically transform preferentially hamster lymphocytes. Thus the hamster lymphocytic leukemia originally induced by the wild-type SV40 was most likely a chance-stochastic event rather than the result of tropism-determinism mediated by the virus, as is usually the case with leukemogenic RNA viruses.

  4. Primordial germ cell-mediated chimera technology produces viable pure-line Houbara bustard offspring: potential for repopulating an endangered species.

    Directory of Open Access Journals (Sweden)

    Ulrich Wernery

    2010-12-01

    Full Text Available The Houbara bustard (Chlamydotis undulata is a wild seasonal breeding bird populating arid sandy semi-desert habitats in North Africa and the Middle East. Its population has declined drastically during the last two decades and it is classified as vulnerable. Captive breeding programmes have, hitherto, been unsuccessful in reviving population numbers and thus radical technological solutions are essential for the long term survival of this species. The purpose of this study was to investigate the use of primordial germ cell-mediated chimera technology to produce viable Houbara bustard offspring.Embryonic gonadal tissue was dissected from Houbara bustard embryos at eight days post-incubation. Subsequently, Houbara tissue containing gonadal primordial germ cells (gPGCs was injected into White Leghorn chicken (Gallus gallus domesticus embryos, producing 83/138 surviving male chimeric embryos, of which 35 chimeric roosters reached sexual maturity after 5 months. The incorporation and differentiation of Houbara gPGCs in chimeric chicken testis were assessed by PCR with Houbara-specific primers and 31.3% (5/16 gonads collected from the injected chicken embryos showed the presence of donor Houbara cells. A total of 302 semen samples from 34 chimeric roosters were analyzed and eight were confirmed as germline chimeras. Semen samples from these eight roosters were used to artificially inseminate three female Houbara bustards. Subsequently, 45 Houbara eggs were obtained and incubated, two of which were fertile. One egg hatched as a male live born Houbara; the other was female but died before hatching. Genotyping confirmed that the male chick was a pure-line Houbara derived from a chimeric rooster.This study demonstrates for the first time that Houbara gPGCs can migrate, differentiate and eventually give rise to functional sperm in the chimeric chicken testis. This approach may provide a promising tool for propagation and conservation of endangered avian

  5. Effects of water-filtered infrared-A and of heat on cell death, inflammation, antioxidative potential and of free radical formation in viable skin--first results.

    Science.gov (United States)

    Piazena, Helmut; Pittermann, Wolfgang; Müller, Werner; Jung, Katinka; Kelleher, Debra K; Herrling, Thomas; Meffert, Peter; Uebelhack, Ralf; Kietzmann, Manfred

    2014-09-05

    The effects of water-filtered infrared-A (wIRA) and of convective heat on viability, inflammation, inducible free radicals and antioxidative power were investigated in natural and viable skin using the ex vivo Bovine Udder System (BUS) model. Therefore, skin samples from differently treated parts of the udder of a healthy cow were analyzed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test, by prostaglandin E2 (PGE2) measurement and by electron spin resonance (ESR) spectroscopy. Neither cell viability, the inflammation status, the radical status or the antioxidative defence systems of the skin were significantly affected by wIRA applied within 30 min by using an irradiance of 1900 W m(-2) which is of relevance for clinical use, but which exceeded the maximum solar IR-A irradiance at the Earth's surface more than 5 times and which resulted in a skin surface temperature of about 45 °C without cooling and of about 37 °C with convective cooling by air ventilation. No significant effects on viability and on inflammation were detected when convective heat was applied alone under equivalent conditions in terms of the resulting skin surface temperatures and exposure time. As compared with untreated skin, free radical formation was almost doubled, whereas the antioxidative power was reduced to about 50% after convective heating to about 45 °C. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Of energy and survival incognito: a relationship between viable but non-culturable cells formation and inorganic polyphosphate and formate metabolism in Campylobacter jejuni.

    Science.gov (United States)

    Kassem, Issmat I; Chandrashekhar, Kshipra; Rajashekara, Gireesh

    2013-01-01

    Campylobacter jejuni is a Gram-negative food-borne bacterium that can cause mild to serious diseases in humans. A variety of stress conditions including exposure to formic acid, a weak organic acid, can cause C. jejuni to form viable but non-culturable cells (VBNC), which was proposed as a potential survival mechanism. The inability to detect C. jejuni VBNC using standard culturing techniques may increase the risk of exposure to foods contaminated with this pathogen. However, little is known about the cellular mechanisms and triggers governing VBNC formation. Here, we discuss novel mechanisms that potentially affect VBNC formation in C. jejuni and emphasize the impact of formic acid on this process. Specifically, we highlight findings that show that impairing inorganic polyphosphate (poly-P) metabolism reduces the ability of C. jejuni to form VBNC in a medium containing formic acid. We also discuss the potential effect of poly-P and formate metabolism on energy homeostasis and cognate VBNC formation. The relationship between poly-P metabolism and VBNC formation under acid stress has only recently been identified and may represent a breakthrough in understanding this phenomenon and its impact on food safety.

  7. Failure of RNA synthesis to recover after UV irradiation: an early defect in cells from individuals with Cockayne's syndrome and xeroderma pigmentosum

    Energy Technology Data Exchange (ETDEWEB)

    Mayne, L.V.; Lehmann, A.R.

    1982-04-01

    Previous work has shown that in cells from the ultraviolet-sensitive genetic disorder, Cockayne's syndrome, DNA synthesis fails to recover after ultraviolet irradiation, despite the fact that these cells have no detectable defect in either excision or daughter-strand repair pathways. We now show that Cockayne cells, as well as cells from a number of patients with xeroderma pigmentosum, are sensitive to the lethal effects of UV irradiation in stationary phase under conditions in which no DNA is synthesized after irradiation. Furthermore, in normal and defective human fibroblasts, RNA synthesis is depressed after UV irradiation. In normal (dividing) cells, RNA synthesis recovers very rapidly, but this recovery does not occur in Cockayne cells, and it is reduced or absent in xeroderma pigmentosum cells from different complementation groups. Qualitatively, similar results are obtained with cells in stationary phase. The recovery of RNA synthesis in the various defective cell strains is not correlated with the overall extent of excision repair, but there is some correlation between recovery of RNA synthesis and cell survival after ultraviolet irradiation. These results implicate recovery of RNA synthesis as an important early response to ultraviolet irradiation.

  8. Mononuclear cells from patients recovered from cutaneous leishmaniasis respond to Leishmania major amastigote class I nuclease with a predominant Th1-like response

    Science.gov (United States)

    Farajnia, S; Mahboudi, F; Ajdari, S; Reiner, N E; Kariminia, A; Alimohammadian, M H

    2005-01-01

    The Leishmania major amastigote class I nuclease (LmaCIN) is a developmentally regulated protein that is highly expressed in the amastigote stage of L. major. This protein is homologous to the P4 nuclease of L. pifanoi, which has been shown to induce protective immune response in a murine model. To evaluate LmaCIN as a potential human vaccine candidate, cellular immune responses to recombinant LmaCIN were examined in individuals recovered from Old World cutaneous leishmaniasis. Peripheral blood mononuclear cells (PBMC) from patients recovered from L. major infection were cultured either with recombinant LmaCIN or autoclaved L. major (ALM) as control. rLmaCIN induced significant proliferation of PBMC from 90% of recovered patients. Phenotypic analysis of proliferating cells showed that CD8+ cells were the predominant cell type proliferating in response to rLmaC1N. Screening of culture supernatants for cytokines showed that rLmaCIN induced high levels of interferon (IFN)-γ (mean ± s.e.m.: 1398 ± 179 pg/ml) associated with little interleukin (IL)-10 and little or no IL-5 production. These findings show that LmaCIN is immunogenic in humans during L. major infection and that it can elicit immunological responses relevant to immunoprophylaxis of leishmaniasis. PMID:15730396

  9. Cultured bovine granulosa cells rapidly lose important features of their identity and functionality but partially recover under long-term culture conditions.

    Science.gov (United States)

    Yenuganti, Vengala Rao; Vanselow, Jens

    2017-05-01

    Cell culture models are essential for the detailed study of molecular processes. We analyze the dynamics of changes in a culture model of bovine granulosa cells. The cells were cultured for up to 8 days and analyzed for steroid production and gene expression. According to the expression of the marker genes CDH1, CDH2 and VIM, the cells maintained their mesenchymal character throughout the time of culture. In contrast, the levels of functionally important transcripts and of estradiol and progesterone production were rapidly down-regulated but showed a substantial up-regulation from day 4. FOXL2, a marker for granulosa cell identity, was also rapidly down-regulated after plating but completely recovered towards the end of culture. In contrast, expression of the Sertoli cell marker SOX9 and the lesion/inflammation marker PTGS2 increased during the first 2 days after plating but gradually decreased later on. We conclude that only long-term culture conditions (>4 days) allow the cells to recover from plating stress and to re-acquire characteristic granulosa cell features.

  10. Characterization of cells recovered from the xenotransplanted NG97 human-derived glioma cell line subcultured in a long-term in vitro

    Directory of Open Access Journals (Sweden)

    Heinrich Juliana K

    2008-10-01

    Full Text Available Abstract Background In order to elucidate tumoral progression and drug resistance, cultured cell lines are valuable tools applied on tumor related assays provided they are well established and characterized. Our laboratory settled the NG97 cell line derived from a human astrocytoma grade III, which started to develop and express important phenotypical characteristics of an astrocytoma grade IV after injection in the flank of nude mice. Astrocytomas are extremely aggressive malignancies of the Central Nervous System (CNS and account for 46% of all primary malignant brain tumors. Progression to worse prognosis occurs in 85% of the cases possibly due to changes in cell tumor microenvironment and through biological pathways that are still unclear. Methods This work focused on characterizing the NG97 cell line specifically after being recovered from the xenotransplant, who maintained their undifferentiated characteristics along the following 60th passages in vitro. These cells were subcultivated to evaluate the possible contribution of these undifferentiated characteristics to the malignant progression phenotype. These characteristics were the expression of molecules involved in the processes of migration, dedifferentiation and chromosomal instability. Results Results showed that NG97(ht had an decrease in doubling time through sub cultivation, which was characterized by a converse modulation between the expression of glial fibrillary acidic protein (GFAP and vimentin. In addition, β1 integrins were present in intermediate levels while α5 integrins had a high expression profile as well as fibronectin and laminin. Cytogenetic analysis of NG97(ht revealed several chromosomal abnormalities, 89% of the cells showed to be hyperdiploid and the modal number was assigned to be 63. Several acrocentric chromosomes were visualized and at least 30 figures were attributed to be murine. These findings suggest a possible fusion between the original NG97 cells

  11. An integrated system for synchronous culture of animal cells under controlled conditions

    National Research Council Canada - National Science Library

    Mendoza-Pérez, Elena; Hernández, Vanessa; Palomares, Laura A; Serrato, José A

    2016-01-01

    .... Moreover, only a few viable cells are recovered. Here, we designed an integrated system where a large number of cells from a controlled bioreactor culture is separated by centrifugal elutriation at high viabilities...

  12. Human amniotic fluid stem cells have a potential to recover ovarian function in mice with chemotherapy-induced sterility

    National Research Council Canada - National Science Library

    Dongmei Lai; Fangyuan Wang; Yifei Chen; Li Wang; Yanlin Wang; Weiwei Cheng

    2013-01-01

      Doc number: 34 Abstract Background: Human amniotic fluid cells (hAFCs) may differentiate into multiple cell lineages and thus have a great potential to become a donor cell source for regenerative medicine...

  13. Where Do All the Phytoplankton Go? Challenges in Keeping Track of Viable Cells in Phytoplankton Communities Using Flow Cytometry and Cell Staining

    Science.gov (United States)

    Simmons, L. J.; Fobbe, D. J.; Berges, J. A.

    2016-02-01

    Understanding the dynamics of phytoplankton communities has traditionally focused on differences in growth and related processes among taxa. It is now appreciated that differences in mortality could be equally important in contributing to these dynamics. Studying mortality in communities is difficult, especially on relevant time scales, which could be as short as hours to days. Flow cytometry can potentially provide solutions, because it can allow discrimination of different taxa, and when combined with staining, distinguish live and dead cells. We applied flow cytometry and staining to phytoplankton communities in a model system: a small, well-studied, urban pond in southeastern Wisconsin. Using flow cytometry, it was possible to resolve up to six dominant taxa (most stain also affected other fluorescence channels, requiring compensation. Correlations of numbers of dead cells with environmental factors (e.g. temperature, nutrient concentrations, irradiance) were generally poor, suggesting the greater importance of biotic versus abiotic variables in community mortality dynamics. Ongoing work is focusing on the effects of viral pathogens, grazing and allelopathic interactions using experimental manipulations and individual-based modeling.

  14. Characterization of Toxoplasma gondii-specific T cells recovered from vitreous fluid of patients with ocular toxoplasmosis

    NARCIS (Netherlands)

    E.J. Feron (Eric); V.N. Klaren; E.A. Wierenga (Eddy); A. Kijlstra Prof. Dr; G.M.G.M. Verjans (George)

    2001-01-01

    textabstractPURPOSE: The mechanisms involved in reactivations of latent ocular Toxoplasma gondii (Tg) infections in immunocompetent patients are poorly understood. In view of the possible role of T cells in the immunopathogenesis of the disease, ocular infiltrating T cells

  15. Co-infusion of autologous adipose tissue derived neuronal differentiated mesenchymal stem cells and bone marrow derived hematopoietic stem cells, a viable therapy for post-traumatic brachial plexus injury: A case report

    Directory of Open Access Journals (Sweden)

    Umang G Thakkar

    2014-08-01

    Full Text Available Stem cell therapy is emerging as a viable approach in regenerative medicine. A 31-year-old male with brachial plexus injury had complete sensory-motor loss since 16 years with right pseudo-meningocele at C5-D1 levels and extra-spinal extension up to C7-D1, with avulsion on magnetic resonance imaging and irreversible damage. We generated adipose tissue derived neuronal differentiated mesenchymal stem cells (N-AD-MSC and bone marrow derived hematopoietic stem cells (HSC-BM. Neuronal stem cells expressed β-3 tubulin and glial fibrillary acid protein which was confirmed on immunofluorescence. On day 14, 2.8 ml stem cell inoculum was infused under local anesthesia in right brachial plexus sheath by brachial block technique under ultrasonography guidance with a 1.5-inch-long 23 gauge needle. Nucleated cell count was 2 × 10 4 /μl, CD34+ was 0.06%, and CD45-/90+ and CD45-/73+ were 41.63% and 20.36%, respectively. No untoward effects were noted. He has sustained recovery with re-innervation over a follow-up of 4 years documented on electromyography-nerve conduction velocity study.

  16. Potential of Escherichia coli 0157:H7 to persist and form viable but non-culturable cells on a food-contact surface subjected to cycles of soiling and chemical treatment

    DEFF Research Database (Denmark)

    Marouani-Gadri, Nesrine; Firmesse, Olivier; Chassaing, Danielle

    2010-01-01

    only, a further increase in this proportion occurred 24 h after the CT, suggesting that some of the surviving viable but non-culturable cells finally died. This study shows that conditions leading to E. coli O157:H7 persistence are not likely to arise when good refrigeration and hygiene practices...... no longer detectable after the first week. However, on 66-hour biofilms with 6.7 log CFU/cm², after initially decreasing, E. coli numbers reached 6.6 log CFU/cm² and 8.3 log viable cells/cm² on the 11th day. When E. coli was cultured with a Comamonas testosteroni previously shown to increase E. coli biofilm...

  17. Human amniotic fluid stem cells have a potential to recover ovarian function in mice with chemotherapy-induced sterility.

    Science.gov (United States)

    Lai, Dongmei; Wang, Fangyuan; Chen, Yifei; Wang, Li; Wang, Yanlin; Cheng, Weiwei

    2013-09-04

    Human amniotic fluid cells (hAFCs) may differentiate into multiple cell lineages and thus have a great potential to become a donor cell source for regenerative medicine. The ability of hAFCs to differentiate into germ cell and oocyte-like cells has been previously documented. Herein we report the potential use of hAFCs to help restore follicles in clinical condition involving premature ovarian failure. Human amniotic fluid was obtained via amniocentesis, yielding a subpopulation of cloned hAFCs that was able to form embryoid bodies (EBs) and differentiate into three embryonic germ layers. Moreover, culture of EBs in medium containing human follicular fluid (HFF) or a germ cell maturation factor cocktail (FAC), expressed germ cells markers such as BLIMP1, STELLA, DAZL, VASA, STRA8, SCP3, SCP1, and GDF9. Furthermore, one cell line was grown from clone cells transfected with lentivirus-GFP and displaying morphological characteristics of mesenchymal cells, had the ability to restore ovarian morphology following cell injection into the ovaries of mice sterilized by intraperitoneal injection of cyclophosphamide and busulphan. Restored ovaries displayed many follicle-enclosed oocytes at all stages of development, but no oocytes or follicles were observed in sterilized mice whose ovaries had been injected with medium only (control). Notably, identification of GFP-labeled cells and immunostaining with anti-human antigen-specific antibodies demonstrated that grafted hAFCs survived and differentiated into granulosa cells which directed oocyte maturation. Furthermore, labeling of ovarian tissue for anti-Müllerian hormone expression, a functional marker of folliculogenesis, was strong in hAFCs-transplanted ovaries but inexistent in negative controls. These findings highlight the possibility of using human amniotic fluid-derived stem cells in regenerative medicine, in particular in the area of reproductive health.

  18. Characterization of Toxoplasma gondii-specific T cells recovered from vitreous fluid of patients with ocular toxoplasmosis

    NARCIS (Netherlands)

    Feron, E. J.; Klaren, V. N.; Wierenga, E. A.; Verjans, G. M.; Kijlstra, A.

    2001-01-01

    The mechanisms involved in reactivations of latent ocular Toxoplasma gondii (Tg) infections in immunocompetent patients are poorly understood. In view of the possible role of T cells in the immunopathogenesis of the disease, ocular infiltrating T cells obtained from patients with recurrent ocular

  19. Supplementation of T3 Recovers Hypothyroid Rat Liver Cells from Oxidatively Damaged Inner Mitochondrial Membrane Leading to Apoptosis

    Science.gov (United States)

    Mukherjee, Sutapa; Samanta, Luna; Roy, Anita; Bhanja, Shravani; Chainy, Gagan B. N.

    2014-01-01

    Hypothyroidism is a growing medical concern. There are conflicting reports regarding the mechanism of oxidative stress in hypothyroidism. Mitochondrial oxidative stress is pivotal to thyroid dysfunction. The present study aimed to delineate the effects of hepatic inner mitochondrial membrane dysfunction as a consequence of 6-n-propyl-2-thiouracil-induced hypothyroidism in rats. Increased oxidative stress predominance in the submitochondrial particles (SMP) and altered antioxidant defenses in the mitochondrial matrix fraction correlated with hepatocyte apoptosis. In order to check whether the effects caused by hypothyroidism are reversed by T3, the above parameters were evaluated in a subset of T3-treated hypothyroid rats. Complex I activity was inhibited in hypothyroid SMP, whereas T3 supplementation upregulated electron transport chain complexes. Higher mitochondrial H2O2 levels in hypothyroidism due to reduced matrix GPx activity culminated in severe oxidative damage to membrane lipids. SMP and matrix proteins were stabilised in hypothyroidism but exhibited increased carbonylation after T3 administration. Glutathione content was higher in both. Hepatocyte apoptosis was evident in hypothyroid liver sections; T3 administration, on the other hand, exerted antiapoptotic and proproliferative effects. Hence, thyroid hormone level critically regulates functional integrity of hepatic mitochondria; hypothyroidism injures mitochondrial membrane lipids leading to hepatocyte apoptosis, which is substantially recovered upon T3 supplementation. PMID:24987693

  20. Supplementation of T3 Recovers Hypothyroid Rat Liver Cells from Oxidatively Damaged Inner Mitochondrial Membrane Leading to Apoptosis

    Directory of Open Access Journals (Sweden)

    Sutapa Mukherjee

    2014-01-01

    Full Text Available Hypothyroidism is a growing medical concern. There are conflicting reports regarding the mechanism of oxidative stress in hypothyroidism. Mitochondrial oxidative stress is pivotal to thyroid dysfunction. The present study aimed to delineate the effects of hepatic inner mitochondrial membrane dysfunction as a consequence of 6-n-propyl-2-thiouracil-induced hypothyroidism in rats. Increased oxidative stress predominance in the submitochondrial particles (SMP and altered antioxidant defenses in the mitochondrial matrix fraction correlated with hepatocyte apoptosis. In order to check whether the effects caused by hypothyroidism are reversed by T3, the above parameters were evaluated in a subset of T3-treated hypothyroid rats. Complex I activity was inhibited in hypothyroid SMP, whereas T3 supplementation upregulated electron transport chain complexes. Higher mitochondrial H2O2 levels in hypothyroidism due to reduced matrix GPx activity culminated in severe oxidative damage to membrane lipids. SMP and matrix proteins were stabilised in hypothyroidism but exhibited increased carbonylation after T3 administration. Glutathione content was higher in both. Hepatocyte apoptosis was evident in hypothyroid liver sections; T3 administration, on the other hand, exerted antiapoptotic and proproliferative effects. Hence, thyroid hormone level critically regulates functional integrity of hepatic mitochondria; hypothyroidism injures mitochondrial membrane lipids leading to hepatocyte apoptosis, which is substantially recovered upon T3 supplementation.

  1. Genotoxicity profiles in exfoliated human mammary cells recovered from lactating mothers in Istanbul; relationship with demographic and dietary factors.

    Science.gov (United States)

    Yilmaz, Bayram; Sandal, Suleyman; Ayvaci, Habibe; Tug, Niyazi; Vitrinel, Ayca

    2012-12-12

    We have investigated the presence of DNA damage in human mammary epithelial cells collected from healthy lactating mothers (age, 20-35 years) who were resident in the Istanbul area. Breast milk (10ml) was collected from 30 women between one and two weeks post-partum. Demographic information (parity, breast cancer, occupation, duration of residency in Istanbul, consumption of fish, beef and poultry) was also obtained. Milk samples were diluted 1:1 with RPMI 1640 medium and centrifuged to collect cells. The cells were re-suspended and cell viability was determined by use of 0.4% trypan blue. DNA damage was assessed by use of the comet assay (alkaline single-cell gel electrophoresis). Fifty cells per slide and two slides per sample were scored to evaluate DNA damage. The cells were visually classified into four categories on the basis of extent of migration: undamaged (UD), lightly damaged (LD), moderately damaged (MD) and highly damaged (HD). Total comet scores (TCS) were calculated as: 1× UD+2× LD+3× MD+4× HD. Exfoliated mammary cells of the donors showed high (TCS≥150a.u.), moderate and low DNA damage in 10 (33.3%), 8 (26.7%) and 12 (40%) mothers, respectively. There was no significant correlation between TCS for DNA damage and the duration of previous breastfeeding, parity or age. None of the mothers was vegetarian, smoker or on any medication. Meat and chicken consumption did not significantly correlate with the TCS values. Fish consumption was significantly correlated with TCS results (Spearman's rho=0.39, pmothers in Istanbul may be of dietary origin. Our experience also confirms that sampling breast milk from lactating mothers provides a valuable and non-invasive tool to study DNA damage in mammary cells. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. Rat embryonic hippocampus and induced pluripotent stem cell derived cultured neurons recover from laser-induced subaxotomy.

    Science.gov (United States)

    Selfridge, Aaron; Hyun, Nicholas; Chiang, Chai-Chun; Reyna, Sol M; Weissmiller, April M; Shi, Linda Z; Preece, Daryl; Mobley, William C; Berns, Michael W

    2015-01-01

    Axonal injury and stress have long been thought to play a pathogenic role in a variety of neurodegenerative diseases. However, a model for studying single-cell axonal injury in mammalian cells and the processes of repair has not been established. The purpose of this study was to examine the response of neuronal growth cones to laser-induced axonal damage in cultures of embryonic rat hippocampal neurons and induced pluripotent stem cell (iPSC) derived human neurons. A 532-nm pulsed [Formula: see text] picosecond laser was focused to a diffraction limited spot at a precise location on an axon using a laser energy/power that did not rupture the cell membrane (subaxotomy). Subsequent time series images were taken to follow axonal recovery and growth cone dynamics. After laser subaxotomy, axons thinned at the damage site and initiated a dynamic cytoskeletal remodeling process to restore axonal thickness. The growth cone was observed to play a role in the repair process in both hippocampal and iPSC-derived neurons. Immunofluorescence staining confirmed structural tubulin damage and revealed initial phases of actin-based cytoskeletal remodeling at the damage site. The results of this study indicate that there is a repeatable and cross-species repair response of axons and growth cones after laser-induced damage.

  3. Development of a one-tube extraction and amplification method for DNA analysis of sperm and epithelial cells recovered from forensic samples by laser microdissection.

    Science.gov (United States)

    Meredith, Melanie; Bright, Jo-Anne; Cockerton, Sarah; Vintiner, Sue

    2012-01-01

    Laser microdissection can be used in forensic casework to isolate specific cell types from mixtures of biological samples. Extraction of DNA from selected cells is still required prior to STR amplification. Because of the relatively pristine nature of the recovered cells, laser microdissection is more sensitive than more traditional methods of DNA analysis, theoretically resulting in DNA profiles from less cellular material. A one-tube extraction and amplification method minimises loss of DNA through liquid transfers and reduces the potential for contamination events occurring. In this paper, the development of a one-tube method for the effective extraction of DNA from laser microdissected sperm and epithelial cells is described. The performance of the in-house method was compared to that of a commercial DNA extraction kit for extraction of DNA from sperm and the downstream compatibility with STR amplification was determined for both sperm and epithelial samples. Full Identifiler™ profiles after 28 amplification cycles were obtained from as few as 15 epithelial cells and 30 sperm. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  4. Correspondence of Neutralizing Humoral Immunity and CD4 T Cell Responses in Long Recovered Sudan Virus Survivors.

    Science.gov (United States)

    Sobarzo, Ariel; Stonier, Spencer W; Herbert, Andrew S; Ochayon, David E; Kuehne, Ana I; Eskira, Yael; Fedida-Metula, Shlomit; Tali, Neta; Lewis, Eli C; Egesa, Moses; Cose, Stephen; Lutwama, Julius Julian; Yavelsky, Victoria; Dye, John M; Lobel, Leslie

    2016-05-11

    Robust humoral and cellular immunity are critical for survival in humans during an ebolavirus infection. However, the interplay between these two arms of immunity is poorly understood. To address this, we examined residual immune responses in survivors of the Sudan virus (SUDV) outbreak in Gulu, Uganda (2000-2001). Cytokine and chemokine expression levels in SUDV stimulated whole blood cultures were assessed by multiplex ELISA and flow cytometry. Antibody and corresponding neutralization titers were also determined. Flow cytometry and multiplex ELISA results demonstrated significantly higher levels of cytokine and chemokine responses in survivors with serological neutralizing activity. This correspondence was not detected in survivors with serum reactivity to SUDV but without neutralization activity. This previously undefined relationship between memory CD4 T cell responses and serological neutralizing capacity in SUDV survivors is key for understanding long lasting immunity in survivors of filovirus infections.

  5. Correspondence of Neutralizing Humoral Immunity and CD4 T Cell Responses in Long Recovered Sudan Virus Survivors

    Directory of Open Access Journals (Sweden)

    Ariel Sobarzo

    2016-05-01

    Full Text Available Robust humoral and cellular immunity are critical for survival in humans during an ebolavirus infection. However, the interplay between these two arms of immunity is poorly understood. To address this, we examined residual immune responses in survivors of the Sudan virus (SUDV outbreak in Gulu, Uganda (2000–2001. Cytokine and chemokine expression levels in SUDV stimulated whole blood cultures were assessed by multiplex ELISA and flow cytometry. Antibody and corresponding neutralization titers were also determined. Flow cytometry and multiplex ELISA results demonstrated significantly higher levels of cytokine and chemokine responses in survivors with serological neutralizing activity. This correspondence was not detected in survivors with serum reactivity to SUDV but without neutralization activity. This previously undefined relationship between memory CD4 T cell responses and serological neutralizing capacity in SUDV survivors is key for understanding long lasting immunity in survivors of filovirus infections.

  6. Tumor-selective replication herpes simplex virus-based technology significantly improves clinical detection and prognostication of viable circulating tumor cells

    DEFF Research Database (Denmark)

    Zhang, Wen; Bao, Li; Yang, Shaoxing

    2016-01-01

    Detection of circulating tumor cells remains a significant challenge due to their vast physical and biological heterogeneity. We developed a cell-surface-marker-independent technology based on telomerase-specific, replication-selective oncolytic herpes-simplex-virus-1 that targets telomerase...

  7. Enzymatic isolation of viable human odontoblasts.

    Science.gov (United States)

    Cuffaro, H M; Pääkkönen, V; Tjäderhane, L

    2016-05-01

    To improve an enzymatic method previously used for isolation of rat odontoblasts to isolate viable mature human odontoblasts. Collagenase I, collagenase I/hyaluronidase mixture and hyaluronidase were used to extract mature human odontoblasts from the pulp chamber. Detachment of odontoblasts from dentine was determined with field emission scanning electron microscopy (FESEM) and to analyse the significance of differences in tubular diameter, and the t-test was used. MTT-reaction was used to analyse cell viability, and nonparametric Kruskal-Wallis and Mann-Whitney post hoc tests were used to analyse the data. Immunofluorescent staining of dentine sialoprotein (DSP), aquaporin-4 (AQP4) and matrix metalloproteinase-20 (MMP-20) and quantitative PCR (qPCR) of dentine sialophosphoprotein (DSPP) were used to confirm the odontoblastic nature of the cells. MTT-reaction and FESEM demonstrated collagenase I/hyaluronidase resulted in more effective detachment and higher viability than collagenase I alone. Hyaluronidase alone was not able to detach odontoblasts. Immunofluorescence revealed the typical odontoblastic-morphology with one process, and DSP, AQP4 and MMP-20 were detected. Quantitative PCR of DSPP confirmed that the isolated cells expressed this odontoblast-specific gene. The isolation of viable human odontoblasts was successful. The cells demonstrated morphology typical for odontoblasts and expressed characteristic odontoblast-type genes and proteins. This method will enable new approaches, such as apoptosis analysis, for studies using fully differentiated odontoblasts. © 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  8. Hybrid systems for distributed power generation based on pressurisation and heat recovering of an existing 100 kW molten carbonate fuel cell

    Science.gov (United States)

    Grillo, Olivia; Magistri, Loredana; Massardo, Aristide F.

    In this paper, different pressurisation and heat recovering techniques for an existing 100 kW molten carbonate fuel cell developed by Ansaldo fuel cells (formerly Ansaldo Ricerche) such as electrically driven compressors for anode (fuel) and cathode side (air), turbocharger, simple cycle gas turbine and regenerated gas turbine are analysed and discussed. The analysis has been carried out using for the FCS-MCFC stack simulation a model developed by the Thermochemical Power Group of the University of Genoa carefully tested with available experimental design point data. The design point hybrid system configurations have been analysed in detail using the code HS-MCFC based on the cited MCFC stack model and developed using Simulink language [Master Thesis, University of Genoa, 2001]. The different hybrid systems design point performance are presented and discussed in great detail, taking into account efficiency, specific power, costs, feasibility, and the need of modification of the existing FC-MCFC systems. Due to the size of the hybrid systems investigated (100-150 kW) they are very interesting for distributed power generation applications.

  9. The application of alkaline lysis and pressure cycling technology in the differential extraction of DNA from sperm and epithelial cells recovered from cotton swabs.

    Science.gov (United States)

    Nori, Deepthi V; McCord, Bruce R

    2015-09-01

    This study reports the development of a two-step protocol using pressure cycling technology (PCT) and alkaline lysis for differential extraction of DNA from mixtures of sperm and vaginal epithelial cells recovered from cotton swabs. In controlled experiments, in which equal quantities of sperm and female epithelial cells were added to cotton swabs, 5 min of pressure pulsing in the presence of 0.4 M NaOH resulted in 104 ± 6% recovery of female epithelial DNA present on the swab. Following the pressure treatment, exposing the swabs to a second 5-min alkaline treatment at 95 °C without pressure resulted in the selective recovery of 69 ± 6% of the sperm DNA. The recovery of the vaginal epithelia and sperm DNA was optimized by examining the effect of sodium hydroxide concentration, incubation temperature, and time. Following the alkaline lysis steps, the samples were neutralized with 2 M Tris (pH 7.5) and purified with phenol-chloroform-isoamyl alcohol to permit downstream analysis. The total processing time to remove both fractions from the swab was less than 20 min. Short tandem repeat (STR) analysis of these fractions obtained from PCT treatment and alkaline lysis generated clean profiles of female epithelial DNA and male sperm DNA for 1:1 mixtures of female and male cells and predominant male profiles for mixtures up to 5:1 female to male cells. By reducing the time and increasing the recovery of DNA from cotton swabs, this new method presents a novel and potentially useful procedure for forensic differential extractions.

  10. The binding of rhBMP-2 to the receptors of viable MC3T3-E1 cells and the question of cooperativity

    Energy Technology Data Exchange (ETDEWEB)

    Wiemann, M.; Rumpf, H.M.; Bingmann, D.; Jennissen, H.P. [Universitaetsklinikum Essen (Germany). Inst. fuer Physiologie; Universitaetsklinikum Essen (Germany). Inst. fuer Physiologiesche Chemie

    2001-12-01

    The binding of rhBMP-2 to its receptors, the signal transduction cascade and the final responses of bone cells, osteoprogenitor cells and derived cell lines is of high fundamental and clinical interest. In this report concentration-response curves of the osteoblast cell line MC3T3-E1 under influence of rhBMP-2 was investigated. The biological response of the cells (corresponding to a down-stream effect of the receptor state-function) was monitored in pilot experiments by the MC3T3-cell alkaline phosphatase-induction test (MC3T3-cell ALP-induction test). It is shown that the MC3T3-cell ALP-induction test is a good tool for measuring biologically active recombinant human BMP-2 (rhBMP-2) in crude extracts of E. coli as well as in highly purified form. In addition this test is very sensitive to chemically induced structural changes of rhBMP-2 such as those resulting from a radiolabeling of rhBMP-2 by the Bolton-Hunter procedure. The latter procedure reduces the biological activity of rhBMP-2 by a factor of 3-4. The measured concentration-response curves could all be non-linearly fitted to a rectangular hyperbola. The half-maximal saturation, K{sub 0.5}, is found between 30-100 nM rhBMP-2 (= 0.8-2.5 {mu}g/ml). The effect of rhBMP-2 shows a plateau i.e. maximal response at ca. 300-1000 nM rhBMP-2 (= 8-25 {mu}g/ml). The data thus indicate a non-cooperative binding-response behavior. This was unexpected since BMP-2 binds simultaneously to two cooperating receptors of type 1 and type 2. However in the very low concentration range of employed rhBMP-2 a variable response of the cells was measured so that a full exclusion of cooperativity cannot be concluded at the present time. This will be clarified by future experiments. (orig.)

  11. Enumeration of viable and non-viable larvated Ascaris eggs with quantitative PCR.

    Science.gov (United States)

    Raynal, Maria; Villegas, Eric N; Nelson, Kara L

    2012-12-01

    The goal of this study was to further develop an incubation-quantitative polymerase chain reaction (qPCR) method for quantifying viable Ascaris eggs by characterizing the detection limit and number of template copies per egg, determining the specificity of the method, and testing the method with viable and inactivated larvated eggs. The number of template copies per cell was determined by amplifying DNA from known numbers of eggs at different development stages; the value was estimated to be 32 copies. The specificity of the method was tested against a panel of bacteria, fungi, protozoa and helminths, and no amplification was found with non-target DNA. Finally, fully larvated eggs were inactivated by four different treatments: 254 nm ultraviolet light, 2,000 ppm NH(3)-N at pH 9, moderate heat (48 °C) and high heat (70 °C). Concentrations of treated eggs were measured by direct microscopy and incubation-qPCR. The qPCR signal decreased following all four treatments, and was in general agreement with the decrease in viable eggs determined by microscopy. The incubation-qPCR method for enumerating viable Ascaris eggs is a promising approach that can produce results faster than direct microscopy, and may have benefits for applications such as assessing biosolids.

  12. The Chemically Synthesized Ageladine A-Derivative LysoGlow84 Stains Lysosomes in Viable Mammalian Brain Cells and Specific Structures in the Marine Flatworm Macrostomum lignano

    Directory of Open Access Journals (Sweden)

    Thorsten Mordhorst

    2015-02-01

    Full Text Available Based on the chemical structure and the known chemical synthesis of the marine sponge alkaloid ageladine A, we synthesized the ageladine A-derivative 4-(naphthalene-2-yl-1H-imidazo[4,5-c]pyridine trifluoroacetate (LysoGlow84. The two-step synthesis started with the Pictet-Spengler reaction of histamine and naphthalene-2-carbaldehyde to a tetrahydropyridine intermediate, which was dehydrogenated with activated manganese (IV oxide to LysoGlow84. Structure and purity of the synthesized LysoGlow84 were confirmed by NMR spectroscopy and mass spectrometry. The fluorescence intensity emitted by LysoGlow84 depended strongly on the pH of the solvent with highest fluorescence intensity recorded at pH 4. The fluorescence maximum (at 315 nm excitation was observed at 440 nm. Biocompatibility of LysoGlow84 was investigated using cultured rat brain astrocytes and the marine flatworm Macrostomum lignano. Exposure of the astrocytes for up to 6 h to micromolar concentrations of LysoGlow84 did not compromise cell viability, as demonstrated by several viability assays, but revealed a promising property of this compound for staining of cellular vesicles. Conventional fluorescence microscopy as well as confocal scanning microscopy of LysoGlow84-treated astrocytes revealed co-localization of LysoGlow84 fluorescence with that of LysoTracker® Red DND-99. LysoGlow84 stained unclear structures in Macrostomum lignano, which were identified as lysosomes by co-staining with LysoTracker. Strong fluorescence staining by LysoGlow84 was further observed around the worms’ anterior gut and the female genital pore which were not counterstained by LysoTracker Red. Thus, LysoGlow84 is a new promising dye that stains lysosomes and other acidic compartments in cultured cells and in worms.

  13. The chemically synthesized ageladine A-derivative LysoGlow84 stains lysosomes in viable mammalian brain cells and specific structures in the marine flatworm Macrostomum lignano.

    Science.gov (United States)

    Mordhorst, Thorsten; Awal, Sushil; Jordan, Sebastian; Petters, Charlotte; Sartoris, Linda; Dringen, Ralf; Bickmeyer, Ulf

    2015-02-11

    Based on the chemical structure and the known chemical synthesis of the marine sponge alkaloid ageladine A, we synthesized the ageladine A-derivative 4-(naphthalene-2-yl)-1H-imidazo[4,5-c]pyridine trifluoroacetate (LysoGlow84). The two-step synthesis started with the Pictet-Spengler reaction of histamine and naphthalene-2-carbaldehyde to a tetrahydropyridine intermediate, which was dehydrogenated with activated manganese (IV) oxide to LysoGlow84. Structure and purity of the synthesized LysoGlow84 were confirmed by NMR spectroscopy and mass spectrometry. The fluorescence intensity emitted by LysoGlow84 depended strongly on the pH of the solvent with highest fluorescence intensity recorded at pH 4. The fluorescence maximum (at 315 nm excitation) was observed at 440 nm. Biocompatibility of LysoGlow84 was investigated using cultured rat brain astrocytes and the marine flatworm Macrostomum lignano. Exposure of the astrocytes for up to 6 h to micromolar concentrations of LysoGlow84 did not compromise cell viability, as demonstrated by several viability assays, but revealed a promising property of this compound for staining of cellular vesicles. Conventional fluorescence microscopy as well as confocal scanning microscopy of LysoGlow84-treated astrocytes revealed co-localization of LysoGlow84 fluorescence with that of LysoTracker® Red DND-99. LysoGlow84 stained unclear structures in Macrostomum lignano, which were identified as lysosomes by co-staining with LysoTracker. Strong fluorescence staining by LysoGlow84 was further observed around the worms' anterior gut and the female genital pore which were not counterstained by LysoTracker Red. Thus, LysoGlow84 is a new promising dye that stains lysosomes and other acidic compartments in cultured cells and in worms.

  14. Viable Syntax: Rethinking Minimalist Architecture

    Directory of Open Access Journals (Sweden)

    Ken Safir

    2010-03-01

    Full Text Available Hauser et al. (2002 suggest that the human language faculty emerged as a genetic innovation in the form of what is called here a ‘keystone factor’—a single, simple, formal mental capability that, interacting with the pre-existing faculties of hominid ancestors, caused a cascade of effects resulting in the language faculty in modern humans. They take Merge to be the keystone factor, but instead it is posited here that Merge is the pre-existing mechanism of thought made viable by a principle that permits relations interpretable at the interfaces to be mapped onto c-command. The simplified minimalist architecture proposed here respects the keystone factor as closely as possible, but is justified on the basis of linguistic analyses it makes available, including a relativized intervention theory applicable across Case, scope, agreement, selection and linearization, a derivation of the A/A’-distinction from Case theory, and predictions such as why in situ wh-interpretation is island-insensitive, but susceptible to intervention effects.

  15. Experimental design for the optimization of propidium monoazide treatment to quantify viable and non-viable bacteria in piggery effluents.

    Science.gov (United States)

    Desneux, Jérémy; Chemaly, Marianne; Pourcher, Anne-Marie

    2015-08-16

    Distinguishing between viable and dead bacteria in animal and urban effluents is a major challenge. Among existing methods, propidium monoazide (PMA)-qPCR is a promising way to quantify viable cells. However, its efficiency depends on the composition of the effluent, particularly on total suspended solids (TSS)) and on methodological parameters. The aim of this study was evaluate the influence of three methodological factors (concentration of PMA, incubation time and photoactivation time) on the efficiency of PMA-qPCR to quantify viable and dead cells of Listeria monocytogenes used as a microorganism model, in two piggery effluents (manure and lagoon effluent containing 20 and 0.4 TSS g.kg(-1), respectively). An experimental design strategy (Doehlert design and desirability function) was used to identify the experimental conditions to achieve optimal PMA-qPCR results. The quantification of viable cells of L. monocytogenes was mainly influenced by the concentration of PMA in the manure and by the duration of photoactivation in the lagoon effluent. Optimal values differed with the matrix: 55 μM PMA, 5 min incubation and 56 min photoactivation for manure and 20 μM PMA, 20 min incubation and 30 min photoactivation for lagoon effluent. Applied to five manure and four lagoon samples, these conditions resulted in satisfactory quantification of viable and dead cells. PMA-qPCR can be used on undiluted turbid effluent with high levels of TSS, provided preliminary tests are performed to identify the optimal conditions.

  16. Detection of viable toxigenic Vibrio cholerae and virulent Shigella ...

    African Journals Online (AJOL)

    A rapid and sensitive assay was developed for the detection of low numbers of viable Vibrio cholerae and Shigella spp. cells in environmental and drinking water samples. Water samples were filtered, and the filters were enriched in a non-selective medium. The enrichment cultures were prepared for polymerase chain ...

  17. Detection of viable toxigenic Vibrio cholerae and virulent Shigella ...

    African Journals Online (AJOL)

    DRINIE

    2003-04-02

    Apr 2, 2003 ... A rapid and sensitive assay was developed for the detection of low numbers of viable Vibrio cholerae and Shigella spp. cells in environmental and drinking water samples. Water samples were filtered, and the filters were enriched in a non-selective medium. The enrichment cultures were prepared for ...

  18. Strategy for analysis of coatings and subsurface sludge recovered during hot cell examinations of N Reactor elements from Hanford K Basins

    Energy Technology Data Exchange (ETDEWEB)

    Makenas, B.J. [Duke Engineering and Services Hanford, Inc., Richland, WA (United States)

    1997-09-22

    Subsurface sludge and/or surface coating material have been collected for four N Reactor fuel elements from K West Basin and one element from K East Basin. It is proposed that examinations of the fairly small volumes of recovered material proceed in order to identify the constituents and their potential impacts on fuel and sludge processing. Lists of potential examination techniques and material allocations are given in this report.

  19. Optimizing Viable Leukocyte Sampling from the Female Genital Tract for Clinical Trials: An International Multi-Site Study

    Science.gov (United States)

    De Rosa, Stephen C.; Martinson, Jeffrey A.; Plants, Jill; Brady, Kirsten E.; Gumbi, Pamela P.; Adams, Devin J.; Vojtech, Lucia; Galloway, Christine G.; Fialkow, Michael; Lentz, Gretchen; Gao, Dayong; Shu, Zhiquan; Nyanga, Billy; Izulla, Preston; Kimani, Joshua; Kimwaki, Steve; Bere, Alfred; Moodie, Zoe; Landay, Alan L.; Passmore, Jo-Ann S.; Kaul, Rupert; Novak, Richard M.; McElrath, M. Juliana; Hladik, Florian

    2014-01-01

    Background Functional analysis of mononuclear leukocytes in the female genital mucosa is essential for understanding the immunologic effects of HIV vaccines and microbicides at the site of HIV exposure. However, the best female genital tract sampling technique is unclear. Methods and Findings We enrolled women from four sites in Africa and the US to compare three genital leukocyte sampling methods: cervicovaginal lavages (CVL), endocervical cytobrushes, and ectocervical biopsies. Absolute yields of mononuclear leukocyte subpopulations were determined by flow cytometric bead-based cell counting. Of the non-invasive sampling types, two combined sequential cytobrushes yielded significantly more viable mononuclear leukocytes than a CVL (pbiopsies. Sample yields were consistent between sites. In a subgroup analysis, we observed significant reproducibility between replicate same-day biopsies (r = 0.89, p = 0.0123). Visible red blood cells in cytobrushes increased leukocyte yields more than three-fold (p = 0.0078), but did not change their subpopulation profile, indicating that these leukocytes were still largely derived from the mucosa and not peripheral blood. We also confirmed that many CD4+ T cells in the female genital tract express the α4β7 integrin, an HIV envelope-binding mucosal homing receptor. Conclusions CVL sampling recovered the lowest number of viable mononuclear leukocytes. Two cervical cytobrushes yielded comparable total numbers of viable leukocytes to one biopsy, but cytobrushes and biopsies were biased toward macrophages and T lymphocytes, respectively. Our study also established the feasibility of obtaining consistent flow cytometric analyses of isolated genital cells from four study sites in the US and Africa. These data represent an important step towards implementing mucosal cell sampling in international clinical trials of HIV prevention. PMID:24454917

  20. Pure Quantum Interpretations Are not Viable

    Science.gov (United States)

    Schmelzer, I.

    2011-02-01

    Pure interpretations of quantum theory, which throw away the classical part of the Copenhagen interpretation without adding new structure to its quantum part, are not viable. This is a consequence of a non-uniqueness result for the canonical operators.

  1. Two missense mutations, E123Q and K151E, identified in the ERG11 allele of an azole-resistant isolate of Candida kefyr recovered from a stem cell transplant patient for acute myeloid leukemia

    Directory of Open Access Journals (Sweden)

    Célia Couzigou

    2014-07-01

    Full Text Available We report on the first cloning and nucleotide sequencing of an ERG11 allele from a clinical isolate of Candida kefyr cross-resistant to azole antifungals. It was recovered from a stem cell transplant patient, in an oncohematology unit exhibiting unexpected high prevalence of C. kefyr. Two amino acid substitutions were identified: K151E, whose role in fluconazole resistance was already demonstrated in Candida albicans, and E123Q, a new substitution never described so far in azole-resistant Candida yeast.

  2. DEMONSTRATION OF FUEL CELLS TO RECOVER ENERGY FROM LANDFILL GAS - PHASE III. DEMONSTRATION TESTS - PHASE IV. GUIDELINES AND DEMONSTRATIONS - VOLUME 2. APPENDICES

    Science.gov (United States)

    The report summarizes the results of a four-phase program to demonstrate that fuel cell energy recovery using a commercial phosphoric acid fuel cell is both environmentally sound and commercially feasible. Phase I, a conceptual design and evaluation study, addressed the technical...

  3. Dichotomy of the human T cell response to Leishmania antigens. I. Th1-like response to Leishmania major promastigote antigens in individuals recovered from cutaneous leishmaniasis

    DEFF Research Database (Denmark)

    Kemp, M; Hey, A S; Kurtzhals, J A

    1994-01-01

    The T cell response to antigens from Leishmania major promastigotes was investigated in peripheral blood mononuclear cells from Sudanese individuals with a history of cutaneous leishmaniasis (CL), Sudanese individuals with positive DTH reaction in the leishmanin skin test but with no history...

  4. DEMONSTRATION OF FUEL CELLS TO RECOVER ENERGY FROM LANDFILL GAS - PHASE III. DEMONSTRATION TESTS - PHASE IV. GUIDELINES AND RECOMMENDATIONS- VOLUME 1. TECHNICAL REPORT

    Science.gov (United States)

    The report summarizes the results of a four-phase program to demonstrate that fuel cell energy recovery using a commercial phosphoric acid fuel cell is both environmentally sound and commercially feasible. Phase I, a conceptual design and evaluation study, addressed the technical...

  5. Grazing of particle-associated bacteria-an elimination of the non-viable fraction.

    Science.gov (United States)

    Gonsalves, Maria-Judith; Fernandes, Sheryl Oliveira; Priya, Madasamy Lakshmi; LokaBharathi, Ponnapakkam Adikesavan

    Quantification of bacteria being grazed by microzooplankton is gaining importance since they serve as energy subsidies for higher trophic levels which consequently influence fish production. Hence, grazing pressure on viable and non-viable fraction of free and particle-associated bacteria in a tropical estuary controlled mainly by protist grazers was estimated using the seawater dilution technique. In vitro incubations over a period of 42h showed that at the end of 24h, growth coefficient (k) of particle-associated bacteria was 9 times higher at 0.546 than that of free forms. Further, 'k' value of viable cells on particles was double that of free forms at 0.016 and 0.007, respectively. While bacteria associated with particles were grazed (coefficient of removal (g)=0.564), the free forms were relatively less grazed indicating that particle-associated bacteria were exposed to grazers in these waters. Among the viable and non-viable forms, 'g' of non-viable fraction (particle-associated bacteria=0.615, Free=0.0086) was much greater than the viable fraction (particle-associated bacteria=0.056, Free=0.068). Thus, grazing on viable cells was relatively low in both the free and attached states. These observations suggest that non-viable forms of particle-associated bacteria were more prone to grazing and were weeded out leaving the viable cells to replenish the bacterial standing stock. Particle colonization could thus be a temporary refuge for the "persistent variants" where the viable fraction multiply and release their progeny. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  6. Using Generic Examples to Make Viable Arguments

    Science.gov (United States)

    Adams, Anne E.; Ely, Rob; Yopp, David

    2017-01-01

    The twenty-first century has seen an increased call to train students to craft mathematical arguments. The third of the Common Core's (CCSS) Standards for Mathematical Practice (SMP 3) (CCSSI 2010) calls for all mathematically proficient students to "construct viable arguments" to support the truth of their ideas and to "critique…

  7. Recover heat from waste incineration

    Energy Technology Data Exchange (ETDEWEB)

    Ganapathy, V. [ABCO Industries, Abilene, TX (United States)

    1995-09-01

    Using these guidelines, engineers can address critical design problems associated with burning process-waste streams and select cost-effective waste-heat boilers. Incinerating contaminant streams is a win-win situation: (1) complete destruction of pollutant(s) is attained and (2) valuable thermal energy is recovered as steam and returned to process, thus conserving energy. However, recovering thermal energy from incinerated flue-gas streams contains some caveats. This treatment method creates a large high-temperature flue gas from which valuable thermal energy is recovered as saturated or superheated steam. Unfortunately, because a process-waste stream is used as feed, this stream will have variations in contaminant and component concentrations which influence the load on the boiler. Also, burning contaminants may create acid gases which will accelerate corrosion problems for the boiler at elevated temperatures. The following guidelines and checklist clarify the do`s and don`ts when designing waste-heat boilers.

  8. Recirculated and Energy Recovered Linacs

    CERN Document Server

    Geoffr-Ey-Kraff

    2003-01-01

    Linacs that are recirculated share many characteristics with ordinary linacs, including the ability to accelerate electron beams FR-om an injector to high energy with relatively little (normalized) emittance growth and the ability to deliver ultrashort bunch duration pulses to users. When such linacs are energy recovered, the additional possibility of accelerating very high average beam current arises. Because this combination of beam properties is not possible FR-om either a conventional linac, or FR-om storage rings where emittance and pulse length are set by the equilibrium between radiation damping and quantum excitation of oscillations about the closed orbit, energy recovered linacs are being considered for an increasing variety of applications. These possibilities extend FR-om high power FR-ee-electron lasers and recirculated linac light sources, to electron coolers for high energy colliders or actual electron-ion colliding- beam machines based on an energy recovered linac for the electrons.

  9. CRISPR/Cas9-Mediated Deletion of CTG Expansions Recovers Normal Phenotype in Myogenic Cells Derived from Myotonic Dystrophy 1 Patients.

    Science.gov (United States)

    Provenzano, Claudia; Cappella, Marisa; Valaperta, Rea; Cardani, Rosanna; Meola, Giovanni; Martelli, Fabio; Cardinali, Beatrice; Falcone, Germana

    2017-12-15

    Myotonic dystrophy type 1 (DM1) is the most common adult-onset muscular dystrophy, characterized by progressive myopathy, myotonia, and multi-organ involvement. This dystrophy is an inherited autosomal dominant disease caused by a (CTG)n expansion within the 3' untranslated region of the DMPK gene. Expression of the mutated gene results in production of toxic transcripts that aggregate as nuclear foci and sequester RNA-binding proteins, resulting in mis-splicing of several transcripts, defective translation, and microRNA dysregulation. No effective therapy is yet available for treatment of the disease. In this study, myogenic cell models were generated from myotonic dystrophy patient-derived fibroblasts. These cells exhibit typical disease-associated ribonuclear aggregates, containing CUG repeats and muscleblind-like 1 protein, and alternative splicing alterations. We exploited these cell models to develop new gene therapy strategies aimed at eliminating the toxic mutant repeats. Using the CRISPR/Cas9 gene-editing system, the repeat expansions were removed, therefore preventing nuclear foci formation and splicing alterations. Compared with the previously reported strategies of inhibition/degradation of CUG expanded transcripts by various techniques, the advantage of this approach is that affected cells can be permanently reverted to a normal phenotype. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  10. Leishmania donovani-reactive Th1- and Th2-like T-cell clones from individuals who have recovered from visceral leishmaniasis

    DEFF Research Database (Denmark)

    Kemp, M; Kurtzhals, J A; Bendtzen, K

    1993-01-01

    Infections in humans by Leishmania donovani parasites can result in a fatal disease, visceral leishmaniasis (VL), or in a self-limiting asymptomatic infection. In murine models of the infection employing Leishmania major, the course of the disease can be directed into a VL-like syndrome by interl......Infections in humans by Leishmania donovani parasites can result in a fatal disease, visceral leishmaniasis (VL), or in a self-limiting asymptomatic infection. In murine models of the infection employing Leishmania major, the course of the disease can be directed into a VL-like syndrome...... only IFN-gamma. This is the first report of a Th1- and Th2-type response in human leishmaniasis. These results suggest that in analogy with murine models, there is a dichotomy in the human T-cell response to L. donovani infections. Preferential activation of IL-4-producing Th2-like cells may...... be involved in the exacerbation of human VL, whereas activation of IFN-gamma-producing Th1 cells may protect the host from severe disease. Identification of leishmanial antigens activating one or the other type of T cells will be important in the development of vaccines against leishmaniasis....

  11. Removal of viable bacteria and endotoxins by Electro Deionization (EDI).

    Science.gov (United States)

    Harada, Norimitsu; Otomo, Teruo; Watabe, Tomoichi; Ase, Tomonobu; Takemura, Takuto; Sato, Toshio

    2011-09-01

    Viable bacteria and endotoxins in water sometimes cause problems for human health. Endotoxins are major components of the outer cell wall of gram-negative bacteria (lipopolysaccharides). In medical procedures, especially haemodialysis (HD) and related therapies (haemodiafiltration (HDF), haemofiltration (HF)), endotoxins in the water for haemodialysis can permeate through the haemodialysis membrane and cause microinflammation or various haemodialysis-related illnesses. To decrease such a biological risk, RO and UF membranes are generally used. Also, hot water disinfection or the chemical disinfection is regularly executed to kill bacteria which produce endotoxins. However, simple treatment methods and equipment may be able to decrease the biological risk more efficiently. In our experiments, we confirmed that viable bacteria and endotoxins were removed by Electro Deionization (EDI) technology and also clarified the desorption mechanisms.

  12. Ventilation and Heat Recovering System

    Directory of Open Access Journals (Sweden)

    Olga Bancea

    2007-01-01

    Full Text Available Some aspects concerning the heat reducing for ventilation, achieved by using heat recovering components and a combined heating--ventilating system, assuring both comfort and human health are presented. The floor imbedded systems together with air outlets elements could fulfill all the aesthetically desires, as well as comfort and they are ideal for family houses.

  13. Smoking Cessation in Recovering Alcoholics

    Science.gov (United States)

    ... Bracing: What Works?Mind/Body Connection: How Your Emotions Affect Your HealthLow-purine DietHealthy Ways to Gain Weight If You’re Underweight Home Your Health Resources Healthcare Management Self-care Smoking Cessation in Recovering Alcoholics Smoking ...

  14. Glomeromycota recovered from cacao soil

    Directory of Open Access Journals (Sweden)

    Kartini - Kramadibrata

    2009-12-01

    Full Text Available KRAMADIBRATA, K. 2009. Glomeromycota recovered from cacao soil. Reinwardtia 12(5: 357–371. ⎯ Glomeromycotan fungi were studied from several cacao plantations in Indonesia (Java and Bali and Ecuador. The identity of 28 species of Glomeromycota associated with cacao is presented.

  15. Survival of Erwinia amylovora in mature apple fruit calyces through the viable but nonculturable (VBNC) state.

    Science.gov (United States)

    Ordax, M; Biosca, E G; Wimalajeewa, S C; López, M M; Marco-Noales, E

    2009-07-01

    Survival of Erwinia amylovora, causal agent of fire blight in pome fruits and other rosaceous plants, was monitored inside mature apples calyces under some storage conditions utilized in fruit. Apple fruit calyces inoculated with two E. amylovora strains and their respective GFP-marked strains were maintained at 26 degrees and 5 degrees C, and the effect of copper treatment was assayed at 0.01 and 0.1 mmol l(-1) CuSO4. In nontreated apples at 26 degrees C, part of the population of E. amylovora survived in the 'viable but nonculturable' (VBNC) state, whereas at 5 degrees C the majority of the population retained culturability. In copper-treated apples, the whole population adopted the VBNC state irrespective of temperature. Regardless of temperature, copper and inoculum dose, VBNC cells recovered culturability and pathogenicity in King's B broth or by host plant passage. Erwinia amylovora survived for at least 35 days in mature apple calyces. Besides, the ability of the pathogen in the VBNC state to regain culturability and pathogenicity suggests that the apple fruit could be a potential carrier of E. amylovora contributing to the spreading of fire blight disease. The risk of E. amylovora dissemination through mature fruit transport, although low, has been demonstrated, and should be considered in pest risk assessments.

  16. Decreases in Gap Junction Coupling Recovers Ca2+ and Insulin Secretion in Neonatal Diabetes Mellitus, Dependent on Beta Cell Heterogeneity and Noise.

    Directory of Open Access Journals (Sweden)

    Aleena M Notary

    2016-09-01

    Full Text Available Diabetes is caused by dysfunction to β-cells in the islets of Langerhans, disrupting insulin secretion and glucose homeostasis. Gap junction-mediated electrical coupling between β-cells in the islet plays a major role in coordinating a pulsatile secretory response at elevated glucose and suppressing insulin secretion at basal glucose. Previously, we demonstrated that a critical number of inexcitable cells can rapidly suppress the overall islet response, as a result of gap junction coupling. This was demonstrated in a murine model of Neonatal Diabetes Mellitus (NDM involving expression of ATP-insensitive KATP channels, and by a multi-cellular computational model of islet electrical activity. Here we examined the mechanisms by which gap junction coupling contributes to islet dysfunction in NDM. We first verified the computational model against [Ca2+] and insulin secretion measurements in islets expressing ATP-insensitive KATP channels under different levels of gap junction coupling. We then applied this model to predict how different KATP channel mutations found in NDM suppress [Ca2+], and the role of gap junction coupling in this suppression. We further extended the model to account for stochastic noise and insulin secretion dynamics. We found experimentally and in the islet model that reductions in gap junction coupling allow progressively greater glucose-stimulated [Ca2+] and insulin secretion following expression of ATP-insensitive KATP channels. The model demonstrated good correspondence between suppression of [Ca2+] and clinical presentation of different NDM mutations. Significant recoveries in [Ca2+] and insulin secretion were predicted for many mutations upon reductions in gap junction coupling, where stochastic noise played a significant role in the recoveries. These findings provide new understanding how the islet functions as a multicellular system and for the role of gap junction channels in exacerbating the effects of decreased

  17. Monotone viable trajectories for functional differential inclusions

    Science.gov (United States)

    Haddad, Georges

    This paper is a study on functional differential inclusions with memory which represent the multivalued version of retarded functional differential equations. The main result gives a necessary and sufficient equations. The main result gives a necessary and sufficient condition ensuring the existence of viable trajectories; that means trajectories remaining in a given nonempty closed convex set defined by given constraints the system must satisfy to be viable. Some motivations for this paper can be found in control theory where F( t, φ) = { f( t, φ, u)} uɛU is the set of possible velocities of the system at time t, depending on the past history represented by the function φ and on a control u ranging over a set U of controls. Other motivations can be found in planning procedures in microeconomics and in biological evolutions where problems with memory do effectively appear in a multivalued version. All these models require viability constraints represented by a closed convex set.

  18. Inhibiting α-synuclein oligomerization by stable cell-penetrating β-synuclein fragments recovers phenotype of Parkinson's disease model flies.

    Directory of Open Access Journals (Sweden)

    Ronit Shaltiel-Karyo

    Full Text Available The intracellular oligomerization of α-synuclein is associated with Parkinson's disease and appears to be an important target for disease-modifying treatment. Yet, to date, there is no specific inhibitor for this aggregation process. Using unbiased systematic peptide array analysis, we identified molecular interaction domains within the β-synuclein polypeptide that specifically binds α-synuclein. Adding such peptide fragments to α-synuclein significantly reduced both amyloid fibrils and soluble oligomer formation in vitro. A retro-inverso analogue of the best peptide inhibitor was designed to develop the identified molecular recognition module into a drug candidate. While this peptide shows indistinguishable activity as compared to the native peptide, it is stable in mouse serum and penetrates α-synuclein over-expressing cells. The interaction interface between the D-amino acid peptide and α-synuclein was mapped by Nuclear Magnetic Resonance spectroscopy. Finally, administering the retro-inverso peptide to a Drosophila model expressing mutant A53T α-synuclein in the nervous system, resulted in a significant recovery of the behavioral abnormalities of the treated flies and in a significant reduction in α-synuclein accumulation in the brains of the flies. The engineered retro-inverso peptide can serve as a lead for developing a novel class of therapeutic agents to treat Parkinson's disease.

  19. Maintenance of pathogenicity during entry into and resuscitation from viable but nonculturable state in Aeromonas hydrophila exposed to natural seawater at low temperature.

    Science.gov (United States)

    Maalej, S; Gdoura, R; Dukan, S; Hammami, A; Bouain, A

    2004-01-01

    To investigate the fate of Aeromonas hydrophila pathogenicity when cells switch, in nutrient-poor filtered sterilized seawater, between the culturable and nonculturable state. Aeromonas hydrophila ATCC 7966, rendered non culturable within 50-55 days of exposure to marine stress conditions, was tested for its ability to maintain haemolysin and to adhere to McCoy cells. Results showed that pathogenicity was lost concomitantly with culturability, whereas cell viability remained undamaged, as determined by the Kogure cell elongation test. However, this loss is only temporary because, following temperature shift from 5 to 23 degrees C, multiple biological activities of recovered Aer. hydrophila cells, which include their ability to lyse human erythrocytes and to attach and destroy McCoy cells were regained. During the temperature-induced resuscitation, constant total cell counts were observed. Moreover, no significant improvement in recovery yield was obtained on brain-heart infusion (BHI) agar plates amended with catalase. We suggest that in addition to the growth of the few undetected culturable cells, there is repair and growth of some mildly injured viable but nonculturable cells. The possibility that nonculturable cells of normally culturable Aer. hydrophila in natural marine environment may constitute a source of infectious diseases posing a public health problem was demonstrated. These experiments may mimic what happens when Aer. hydrophila cells are released in natural seawater with careful attention to the conditions in which surrounding waters gradually become warmer in late summer/early autumn. Copyright 2004 The Society for Applied Microbiology

  20. Strategy for Syntax Error Recovering

    Directory of Open Access Journals (Sweden)

    Henry F Báez

    2003-07-01

    Full Text Available This paper describes a new strategy for syntax error recovering for a compiler that does not have instruction separators like ";" or opening and closing brackets like "{" and "}". This strategy is based on 4 steps. 1. Find a set of tokens (called ACEPTA set for each non terminal symbol of the grammar. 2. During the syntax analysis of each non terminal symbol, eliminate the tokens that are not in the ACEPTA set. 3. Eliminate repeated tokens that are not accepted by the grammar, and 4. Complete symbols in the syntax analysis with the hope that the token that has not been erased later will match with a terminal symbol expected by the syntax analyser; otherwise the symbol will be eliminated in some particular productions. The strategy for sintax error recovering is a method that can be used in whatever not ambiguos context free grammar includying those that use instruction separators like ";". It is implemented with an algorithm and it is much more easy to implement than other strategies for syntax error recovering like those based on stacks.

  1. Air-spore in Cartagena, Spain: viable and non-viable sampling methods.

    Science.gov (United States)

    Elvira-Rendueles, Belen; Moreno, Jose; Garcia-Sanchez, Antonio; Vergara, Nuria; Martinez-Garcia, Maria Jose; Moreno-Grau, Stella

    2013-01-01

    In the presented study the airborne fungal spores of the semiarid city of Cartagena, Spain, are identified and quantified by means of viable or non-viable sampling methods. Airborne fungal samples were collected simultaneously using a filtration method and a pollen and particle sampler based on the Hirst methodology. This information is very useful for elucidating geographical patterns of hay fever and asthma. The qualitative results showed that when the non-viable methodology was employed, Cladosporium, Ustilago, and Alternaria were the most abundant spores identified in the atmosphere of Cartagena, while the viable methodology showed that the most abundant taxa were: Cladosporium, Penicillium, Aspergillus and Alternaria. The quantitative results of airborne fungal spores identified by the Hirst-type air sampler (non-viable method), showed that Deuteromycetes represented 74% of total annual spore counts, Cladosporium being the major component of the fungal spectrum (62.2%), followed by Alternaria (5.3%), and Stemphylium (1.3%). The Basidiomycetes group represented 18.9% of total annual spore counts, Ustilago (7.1%) being the most representative taxon of this group and the second most abundant spore type. Ascomycetes accounted for 6.9%, Nectria (2.3%) being the principal taxon. Oomycetes (0.2%) and Zygomycestes and Myxomycestes (0.06%) were scarce. The prevailing species define our bioaerosol as typical of dry air. The viable methodology was better at identifying small hyaline spores and allowed for the discrimination of the genus of some spore types. However, non-viable methods revealed the richness of fungal types present in the bioaerosol. Thus, the use of both methodologies provides a more comprehensive characterization of the spore profile.

  2. Parejas viables que perduran en el tiempo

    OpenAIRE

    Juan José Cuervo Rodríguez

    2013-01-01

    El presente artículo científico presenta resultados del proceso llevado a cabo en el proyecto de investigación docente "Mecanismos de autorregulación en parejas viables que perduran en el tiempo". Se soporta en una mirada compleja de la psicología basada en una epistemología de la construcción. En el ámbito metodológico, se inscribe en los estudios de terapia familiar desde una perspectiva de la comunicación humana como un todo integrado. Participaron nueve parejas. Los criterios de inclusión...

  3. A rapid biosensor for viable B. anthracis spores.

    Science.gov (United States)

    Baeumner, Antje J; Leonard, Barbara; McElwee, John; Montagna, Richard A

    2004-09-01

    A simple membrane-strip-based biosensor assay has been combined with a nucleic acid sequence-based amplification (NASBA) reaction for rapid (4 h) detection of a small number (ten) of viable B. anthracis spores. The biosensor is based on identification of a unique mRNA sequence from one of the anthrax toxin genes, the protective antigen ( pag), encoded on the toxin plasmid, pXO1, and thus provides high specificity toward B. anthracis. Previously, the anthrax toxins activator ( atxA) mRNA had been used in our laboratory for the development of a biosensor for the detection of a single B. anthracis spore within 12 h. Changing the target sequence to the pag mRNA provided the ability to shorten the overall assay time significantly. The vaccine strain of B. anthracis (Sterne strain) was used in all experiments. A 500-microL sample containing as few as ten spores was mixed with 500 microL growth medium and incubated for 30 min for spore germination and mRNA production. Thus, only spores that are viable were detected. Subsequently, RNA was extracted from lysed cells, selectively amplified using NASBA, and rapidly identified by the biosensor. While the biosensor assay requires only 15 min assay time, the overall process takes 4 h for detection of ten viable B. anthracis spores, and is shortened significantly if more spores are present. The biosensor is based on an oligonucleotide sandwich-hybridization assay format. It uses a membrane flow-through system with an immobilized DNA probe that hybridizes with the target sequence. Signal amplification is provided when the target sequence hybridizes to a second DNA probe that has been coupled to liposomes encapsulating the dye sulforhodamine B. The amount of liposomes captured in the detection zone can be read visually or quantified with a hand-held reflectometer. The biosensor can detect as little as 1 fmol target mRNA (1 nmol L(-1)). Specificity analysis revealed no cross-reactivity with 11 organisms tested, among them closely

  4. A Structure-Based Assembly Screen of Protein Cage Libraries in Living Cells: Experimentally Repacking a Protein-Protein Interface To Recover Cage Formation in an Assembly-Frustrated Mutant.

    Science.gov (United States)

    Cornell, Thomas A; Ardejani, Maziar S; Fu, Jing; Newland, Stephanie H; Zhang, Yu; Orner, Brendan P

    2018-01-17

    Cage proteins, which assemble into often highly symmetric hollow nanoscale capsules, have great potential in applications as far reaching as drug delivery, hybrid nanomaterial engineering, and catalysis. In addition, they are promising model systems for understanding how cellular nanostructures are constructed through protein-protein interactions, and they are beginning to be used as scaffolds for synthetic biology approaches. Recently, there has been renewed interest in the engineering of protein cages, and in support of these strategies, we have recently described a fluorescence-based assay for protein cage assembly that is specific for certain oligomerization states and symmetry-related protein-protein interfaces. In this work, we expand this assay to living cells and a high-throughput assay for screening protein cage libraries using flow cytometry. As a proof of principle, we apply this technique to the screening of libraries of a double-alanine mutant of the mini-ferritin, DNA-binding protein from starved cells (Dps). This mutant, due to disruption of key protein-protein interactions, is unable to assemble into a cage. Randomization of residues surrounding the double mutation afforded a repacked interface and proteins with recovered cage formation, demonstrating the strength and utility of this approach.

  5. Prospective clinical and radiographic evaluation of an allogeneic bone matrix containing stem cells (Trinity Evolution® Viable Cellular Bone Matrix) in patients undergoing two-level anterior cervical discectomy and fusion.

    Science.gov (United States)

    Peppers, Timothy A; Bullard, Dennis E; Vanichkachorn, Jed S; Stanley, Scott K; Arnold, Paul M; Waldorff, Erik I; Hahn, Rebekah; Atkinson, Brent L; Ryaby, James T; Linovitz, Raymond J

    2017-04-26

    Trinity Evolution® (TE), a viable cellular bone allograft, previously demonstrated high fusion rates and no safety-related concerns after single-level anterior cervical discectomy and fusion (ACDF) procedures. This prospective multicenter clinical study was performed to assess the radiographic and clinical outcomes of TE in subjects undergoing two-level ACDF procedures. In a prospective, multicenter study, 40 subjects that presented with symptomatic cervical degeneration at two adjacent vertebral levels underwent instrumented ACDF using TE autograft substitute in a polyetherethereketone (PEEK) cage. At 12 months, radiographic fusion status was evaluated by dynamic motion plain radiographs and thin cut CT with multiplanar reconstruction by a panel that was blinded to clinical outcome. Fusion success was defined by angular motion (≤4°) and the presence of bridging bone across the adjacent vertebral endplates. Clinical pain and function assessments included the Neck Disability Index (NDI), neck and arm pain as evaluated by visual analog scales (VAS), and SF-36 at both 6 and 12 months. At both 6 and 12 months, all clinical outcome scores (SF-36, NDI, and VAS pain) improved significantly (p Trinity Evolution in Anterior Cervical Disectomy and Fusion (ACDF) NCT00951938.

  6. PMA-Linked Fluorescence for Rapid Detection of Viable Bacterial Endospores

    Science.gov (United States)

    LaDuc, Myron T.; Venkateswaran, Kasthuri; Mohapatra, Bidyut

    2012-01-01

    The most common approach for assessing the abundance of viable bacterial endospores is the culture-based plating method. However, culture-based approaches are heavily biased and oftentimes incompatible with upstream sample processing strategies, which make viable cells/spores uncultivable. This shortcoming highlights the need for rapid molecular diagnostic tools to assess more accurately the abundance of viable spacecraft-associated microbiota, perhaps most importantly bacterial endospores. Propidium monoazide (PMA) has received a great deal of attention due to its ability to differentiate live, viable bacterial cells from dead ones. PMA gains access to the DNA of dead cells through compromised membranes. Once inside the cell, it intercalates and eventually covalently bonds with the double-helix structures upon photoactivation with visible light. The covalently bound DNA is significantly altered, and unavailable to downstream molecular-based manipulations and analyses. Microbiological samples can be treated with appropriate concentrations of PMA and exposed to visible light prior to undergoing total genomic DNA extraction, resulting in an extract comprised solely of DNA arising from viable cells. This ability to extract DNA selectively from living cells is extremely powerful, and bears great relevance to many microbiological arenas.

  7. Mammalian Cell Tissue Culture Techniques.

    Science.gov (United States)

    Phelan, Katy; May, Kristin M

    2016-06-01

    Cultured tissues and cells are used extensively in physiological and pharmacological studies. In vitro cultures provide a means of examining cells and tissues without the complex interactions that would be present if the whole organism were studied. A number of special skills are required in order to preserve the structure, function, behavior, and biology of cells in culture. This unit describes the basic skills required to maintain and preserve cell cultures: maintaining aseptic technique, preparing media with the appropriate characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.

  8. Roots of success: cultivating viable community forestry

    Energy Technology Data Exchange (ETDEWEB)

    MacQueen, Duncan

    2009-05-15

    Is community forestry emerging from the shadows? The evidence shows that locally controlled enterprises can be economically viable, and often build on stronger social and environmental foundations than the big private-sector players. Certainly this is an industry in need of a shakeup. Many forests have become flashpoints where agro-industry, large-scale logging concerns and conservation interests clash, while forest-dependent communities are left out in the cold. Meanwhile, governments – driven by concerns over the climate impacts of deforestation – are having to gear up for legal, sustainable forestry production. Community forestry could be crucial to solving many of these challenges. By building on local core capabilities and developing strategic partnerships, they are forging key new business models that could transform the sector.

  9. Polymerase chain reaction-based discrimination of viable from non-viable Mycoplasma gallisepticum

    Directory of Open Access Journals (Sweden)

    Ching Giap Tan

    2014-02-01

    Full Text Available The present study was based on the reverse transcription polymerase chain reaction (RT-PCR of the 16S ribosomal nucleic acid (rRNA of Mycoplasma for detection of viable Mycoplasma gallisepticum. To determine the stability of M. gallisepticum 16S rRNA in vitro, three inactivation methods were used and the suspensions were stored at different temperatures. The 16S rRNA of M. gallisepticum was detected up to approximately 20–25 h at 37 °C, 22–25 h at 16 °C, and 23–27 h at 4 °C. The test, therefore, could detect viable or recently dead M. gallisepticum (< 20 h. The RT-PCR method was applied during an in vivo study of drug efficacy under experimental conditions, where commercial broiler-breeder eggs were inoculated with M. gallisepticum into the yolk. Hatched chicks that had been inoculated in ovo were treated with Macrolide 1. The method was then applied in a flock of day 0 chicks with naturally acquired vertical transmission of M. gallisepticum, treated with Macrolide 2. Swabs of the respiratory tract were obtained for PCR and RT-PCR evaluations to determine the viability of M. gallisepticum. This study proved that the combination of both PCR and RT-PCR enables detection and differentiation of viable from non-viable M. gallisepticum.

  10. Reverse transcriptase real-time PCR for detection and quantification of viable Campylobacter jejuni directly from poultry faecal samples

    DEFF Research Database (Denmark)

    Bui, Thanh Xuan; Wolff, Anders; Madsen, Mogens

    2012-01-01

    and quantification of viable Campylobacter jejuni directly from chicken faecal samples. The results of this method anda DNA-based quantitative real-time PCR (qPCR) method were compared with those of a bacterial culture method. Using bacterial culture andRT-qPCR methods, viable C. jejuni cells could be detected...

  11. The Conceptual Mechanism for Viable Organizational Learning Based on Complex System Theory and the Viable System Model

    Science.gov (United States)

    Sung, Dia; You, Yeongmahn; Song, Ji Hoon

    2008-01-01

    The purpose of this research is to explore the possibility of viable learning organizations based on identifying viable organizational learning mechanisms. Two theoretical foundations, complex system theory and viable system theory, have been integrated to provide the rationale for building the sustainable organizational learning mechanism. The…

  12. Deletion of ultraconserved elements yields viable mice

    Energy Technology Data Exchange (ETDEWEB)

    Ahituv, Nadav; Zhu, Yiwen; Visel, Axel; Holt, Amy; Afzal, Veena; Pennacchio, Len A.; Rubin, Edward M.

    2007-07-15

    Ultraconserved elements have been suggested to retainextended perfect sequence identity between the human, mouse, and ratgenomes due to essential functional properties. To investigate thenecessities of these elements in vivo, we removed four non-codingultraconserved elements (ranging in length from 222 to 731 base pairs)from the mouse genome. To maximize the likelihood of observing aphenotype, we chose to delete elements that function as enhancers in amouse transgenic assay and that are near genes that exhibit markedphenotypes both when completely inactivated in the mouse as well as whentheir expression is altered due to other genomic modifications.Remarkably, all four resulting lines of mice lacking these ultraconservedelements were viable and fertile, and failed to reveal any criticalabnormalities when assayed for a variety of phenotypes including growth,longevity, pathology and metabolism. In addition more targeted screens,informed by the abnormalities observed in mice where genes in proximityto the investigated elements had been altered, also failed to revealnotable abnormalities. These results, while not inclusive of all thepossible phenotypic impact of the deleted sequences, indicate thatextreme sequence constraint does not necessarily reflect crucialfunctions required for viability.

  13. Is Greenberg's "Macro-Carib" viable?

    Directory of Open Access Journals (Sweden)

    Spike Gildea

    Full Text Available In his landmark work Language in the Americas, Greenberg (1987 proposed that Macro-Carib was one of the major low-level stocks of South America, which together with Macro-Panoan and Macro-Ge-Bororo were claimed to comprise the putative Ge-Pano-Carib Phylum. His Macro-Carib includes the isolates Andoke and Kukura, and the Witotoan, Peba-Yaguan, and Cariban families. Greenberg's primary evidence came from person-marking paradigms in individual languages, plus scattered words from individual languages collected into 79 Macro-Carib 'etymologies' and another 64 Amerind 'etymologies'. The goal of this paper is to re-evaluate Greenberg's Macro-Carib claim in the light of the much more extensive and reliable language data that has become available largely since 1987. Based on full person-marking paradigms for Proto-Cariban, Yagua, Bora and Andoke, we conclude that Greenberg's morphological claims are unfounded. For our lexical comparison, we created lexical lists for Proto-Cariban, Proto-Witotoan, Yagua and Andoke, for both Greenberg's 143 putative etymologies and for the Swadesh 100 list. From both lists, a total of 23 potential cognates were found, but no consonantal correspondences were repeated even once. We conclude that our greatly expanded and improved database does not provide sufficient evidence to convince the skeptic that the Macro-Carib hypothesis is viable

  14. Economically viable large-scale hydrogen liquefaction

    Science.gov (United States)

    Cardella, U.; Decker, L.; Klein, H.

    2017-02-01

    The liquid hydrogen demand, particularly driven by clean energy applications, will rise in the near future. As industrial large scale liquefiers will play a major role within the hydrogen supply chain, production capacity will have to increase by a multiple of today’s typical sizes. The main goal is to reduce the total cost of ownership for these plants by increasing energy efficiency with innovative and simple process designs, optimized in capital expenditure. New concepts must ensure a manageable plant complexity and flexible operability. In the phase of process development and selection, a dimensioning of key equipment for large scale liquefiers, such as turbines and compressors as well as heat exchangers, must be performed iteratively to ensure technological feasibility and maturity. Further critical aspects related to hydrogen liquefaction, e.g. fluid properties, ortho-para hydrogen conversion, and coldbox configuration, must be analysed in detail. This paper provides an overview on the approach, challenges and preliminary results in the development of efficient as well as economically viable concepts for large-scale hydrogen liquefaction.

  15. Recovering Place: On the Agency

    Directory of Open Access Journals (Sweden)

    Joern Langhorst

    2012-12-01

    Full Text Available Based on the author’s ongoing involvement in the recovery after hurricane Katrina in New Orleans, and the Lower Ninth Ward neighbourhood in particular, this paper discusses the implications of alternative strategies and tactics involved in the recovery, restoration and rebuilding of post-disaster landscapes. It emphasises that the recovery and rebuilding of community is as important as physical restoration. The empowerment of disaster-affected communities is posited as a central element, in particular, because disasters often reveal long-standing underlying dysfunctions and uneven development patterns as important factors disproportionally affecting communities that were already victimised and marginalised. Design and planning approaches that gain the trust of local residents can reveal hidden, suppressed and alternative narratives and histories central to understanding the processes leading up to the disaster and to the development of viable and sustainable future scenarios. The active participation of traumatised communities is critical to an inclusive discourse on their future, allowing them to become co-authors of the landscapes and places they inhabit instead of victims of hegemonial agendas that created pre-disaster conditions in the first place. The paper discusses a modest spatial intervention in the Lower Ninth Ward and its impacts as an example of the agency of landscape in processes of cultural change. It discusses the instrumentality of a truly public space in a critical location in asserting the viability of a post-disaster neighbourhood and in changing the discourses on human–environment relationships to facilitate a sustainable future in a landscape shaped by challenging social and environmental dynamics.

  16. Is telomerase a viable target in cancer?

    Science.gov (United States)

    Buseman, C.M.; Wright, W.E.; Shay, J.W.

    2012-01-01

    The ideal cancer treatment would specifically target cancer cells yet have minimal or no adverse effects on normal somatic cells. Telomerase, the ribonucleoprotein reverse transcriptase that maintains the ends of human chromosome, is an attractive cancer therapeutic target for exactly this reason [1]. Telomerase is expressed in more than 85% of cancer cells, making it a nearly universal cancer marker, while the majority of normal somatic cells are telomerase negative. Telomerase activity confers limitless replicative potential to cancer cells, a hallmark of cancer which must be attained for the continued growth that characterizes almost all advanced neoplasms [2]. In this review we will summarize the role of telomeres and telomerase in cancer cells, and how properties of telomerase are being exploited to create targeted cancer therapies including telomerase inhibitors, telomerase-targeted immunotherapies and telomerase-driven virotherapies. A frank and balanced assessment of the current state of telomerase inhibitors with caveats and potential limitations will be included. PMID:21802433

  17. Methods for recovering metals from electronic waste, and related systems

    Science.gov (United States)

    Lister, Tedd E; Parkman, Jacob A; Diaz Aldana, Luis A; Clark, Gemma; Dufek, Eric J; Keller, Philip

    2017-10-03

    A method of recovering metals from electronic waste comprises providing a powder comprising electronic waste in at least a first reactor and a second reactor and providing an electrolyte comprising at least ferric ions in an electrochemical cell in fluid communication with the first reactor and the second reactor. The method further includes contacting the powders within the first reactor and the second reactor with the electrolyte to dissolve at least one base metal from each reactor into the electrolyte and reduce at least some of the ferric ions to ferrous ions. The ferrous ions are oxidized at an anode of the electrochemical cell to regenerate the ferric ions. The powder within the second reactor comprises a higher weight percent of the at least one base metal than the powder in the first reactor. Additional methods of recovering metals from electronic waste are also described, as well as an apparatus of recovering metals from electronic waste.

  18. Parejas viables que perduran en el tiempo

    Directory of Open Access Journals (Sweden)

    Juan José Cuervo Rodríguez

    2013-01-01

    Full Text Available El presente artículo científico presenta resultados del proceso llevado a cabo en el proyecto de investigación docente "Mecanismos de autorregulación en parejas viables que perduran en el tiempo". Se soporta en una mirada compleja de la psicología basada en una epistemología de la construcción. En el ámbito metodológico, se inscribe en los estudios de terapia familiar desde una perspectiva de la comunicación humana como un todo integrado. Participaron nueve parejas. Los criterios de inclusión fueron: cinco o más años de convivencia, participación voluntaria, no presentar (ni haber presentado problemáticas especiales que ameriten intervención psicoterapéutica y la obtención de un porcentaje significativo en el uso de estrategias de comunicación asertiva en la resolución de conflictos. El método general utilizado fue el análisis de la comunicación en tarea de conversación. Los principales hallazgos señalan una estrecha relación entre el contexto de desarrollo de las parejas, la emergencia de códigos comunicacionales propios y la posibilidad de perdurar en el tiempo; también, se resalta el tipo de comunicación asertiva o constructiva, la construcción de valores como el respeto y la aceptación de las diferencias, y el deseo por vivir y construir bienestar común, como elementos constitutivos de su identidad como pareja.

  19. Determination of viable Salmonellae from potable and source water through PMA assisted qPCR.

    Science.gov (United States)

    Singh, Gulshan; Vajpayee, Poornima; Bhatti, Saurabh; Ronnie, Nirmala; Shah, Nimish; McClure, Peter; Shanker, Rishi

    2013-07-01

    Resource constrained countries identified as endemic zones for pathogenicity of Salmonella bear an economic burden due to recurring expenditure on medical treatment. qPCR used for Salmonella detection could not discriminate between viable and nonviable cells. Propidium monoazide (PMA) that selectively penetrates nonviable cells to cross-link their DNA, was coupled with ttr gene specific qPCR for quantifying viable salmonellae in source/potable waters collected from a north Indian city. Source water (raw water for urban potable water supply) and urban potable water exhibited viable salmonellae in the range of 2.1×10(4)-2.6×10(6) and 2-7160CFU/100mL, respectively. Potable water at water works exhibited DNA from dead cells but no viable cells were detected. PMA assisted qPCR could specifically detect low numbers of live salmonellae in Source and potable waters. This strategy can be used in surveillance of urban potable water distribution networks to map contamination points for better microbial risk management. Copyright © 2013 Elsevier Inc. All rights reserved.

  20. Asouzu's Complementary Ontology as a Foundation for a Viable ...

    African Journals Online (AJOL)

    This paper on “Asouzu's Complementary Ontology as a foundation for a viable Ethic of the Environment”, posits that an ethic of the environment can be seen as viable if it considers the whole of reality as ontologically relevant. This point of view would free environmental ethics of anthropocentric bias and its attendant ...

  1. Separation of viable and non-viable tomato (Solanum lycopersicum L.) seeds using single seed near-infrared spectroscopy

    DEFF Research Database (Denmark)

    Shrestha, Santosh; Deleuran, Lise Christina; Gislum, René

    2017-01-01

    -viable tomato seeds of two cultivars using chemometrics. The data exploration were performed by principal component analysis (PCA). Subsequently, viable and non-viable seeds were classified by partial least squares-discriminant analysis (PLS-DA) and interval PLS-DA (iPLS-DA). The indication of clustering...... of viable and non-viable seeds were observed in the PCA of each cultivar and the pooled samples. However, the PCA did not exhibit a pattern of separation among the early, normal and late germinated tomato seeds. The NIR spectral regions of 1160–1170, 1383–1397, 1647–1666, 1860–1884 and 1915–1940 nm were...... identified as important for classification of viable and non-viable tomato seeds by iPLS-DA. The sensitivity i.e. ability to correctly identify the positive samples and specificity i.e. ability to reject the negative samples of the (iPLS-DA) model on identified spectral regions for prediction of viable...

  2. Susceptibility of Some Bacterial Contaminants Recovered from ...

    African Journals Online (AJOL)

    resistant bacteria. Isolates tested in this investigation were recovered from a variety of skin and hair cosmetics at the point of sale. Therefore, these contaminants must have been introduced into these products during the manufacturing process.

  3. Characterization of the Viable but Nonculturable (VBNC State in Saccharomyces cerevisiae.

    Directory of Open Access Journals (Sweden)

    Mohammad Salma

    Full Text Available The Viable But Non Culturable (VBNC state has been thoroughly studied in bacteria. In contrast, it has received much less attention in other microorganisms. However, it has been suggested that various yeast species occurring in wine may enter in VBNC following sulfite stress.In order to provide conclusive evidences for the existence of a VBNC state in yeast, the ability of Saccharomyces cerevisiae to enter into a VBNC state by applying sulfite stress was investigated. Viable populations were monitored by flow cytometry while culturable populations were followed by plating on culture medium. Twenty-four hours after the application of the stress, the comparison between the culturable population and the viable population demonstrated the presence of viable cells that were non culturable. In addition, removal of the stress by increasing the pH of the medium at different time intervals into the VBNC state allowed the VBNC S. cerevisiae cells to "resuscitate". The similarity between the cell cycle profiles of VBNC cells and cells exiting the VBNC state together with the generation rate of cells exiting VBNC state demonstrated the absence of cellular multiplication during the exit from the VBNC state. This provides evidence of a true VBNC state. To get further insight into the molecular mechanism pertaining to the VBNC state, we studied the involvement of the SSU1 gene, encoding a sulfite pump in S. cerevisiae. The physiological behavior of wild-type S. cerevisiae was compared to those of a recombinant strain overexpressing SSU1 and null Δssu1 mutant. Our results demonstrated that the SSU1 gene is only implicated in the first stages of sulfite resistance but not per se in the VBNC phenotype. Our study clearly demonstrated the existence of an SO2-induced VBNC state in S. cerevisiae and that the stress removal allows the "resuscitation" of VBNC cells during the VBNC state.

  4. Non-viable Borrelia burgdorferi induce inflammatory mediators and apoptosis in human oligodendrocytes.

    Science.gov (United States)

    Parthasarathy, Geetha; Fevrier, Helene B; Philipp, Mario T

    2013-11-27

    In previous studies, exposure to live Borrelia burgdorferi was shown to induce inflammation and apoptosis of human oligodendrocytes. In this study we assessed the ability of non-viable bacteria (heat killed or sonicated) to induce inflammatory mediators and cell death. Both heat-killed and sonicated bacteria induced release of CCL2, IL-6, and CXCL8 from oligodendrocytes in a dose dependent manner. In addition, non-viable B. burgdorferi also induced cell death as evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and another cell viability assay. These results suggest that spirochetal residues left after bacterial demise, due to treatment or otherwise, may continue to be pathogenic to the central nervous system. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  5. Method for recovering elemental silicon from cutting remains.

    OpenAIRE

    Ulset, Torgeir; Julrud, Stein; Cassayre, Laurent; Chamelot, Pierre; Massot, Laurent; Taxil, Pierre

    2008-01-01

    This invention relates to a method for recovering elemental silicon cutting remains containing silicon particles, wherein the method comprises manufacturing solid anodes from the cutting remains, arranging one or more manufactured anode (s) in an electrolytic cell with a molten salt electrolyte and one or more cathode (s), and applying a potential difference between the one or more anode (s) and cathode (s) to obtain an oxidation of metallic silicon in the one or more anode (s), tran...

  6. Succinic acid production with metabolically engineered E. coli recovered from two-stage fermentation.

    Science.gov (United States)

    Ma, Jiang-Feng; Jiang, Min; Chen, Ke-Quan; Xu, Bing; Liu, Shu-Wen; Wei, Ping; Ying, Han-Jie

    2010-10-01

    Escherichia coli AFP111 cells recovered from spent two-stage fermentation broth were investigated for additional production of succinic acid under anaerobic conditions. Recovered cells produced succinic acid in an aqueous environment with no nutrient supplementation except for glucose and MgCO(3). In addition, initial glucose concentration and cell density had a significant influence on succinic acid mass yield and productivity. Although the final concentration of succinic acid from recovered cells was lower than from two-stage fermentation, an average succinic acid mass yield of 0.85 g/g was achieved with an average productivity of 1.81 g/l h after three rounds of recycling, which was comparable to two-stage fermentation. These results suggested that recovered cells might be reused for the efficient production of succinic acid.

  7. PMA-PhyloChip DNA Microarray to Elucidate Viable Microbial Community Structure

    Science.gov (United States)

    Venkateswaran, Kasthuri J.; Stam, Christina N.; Andersen, Gary L.; DeSantis, Todd

    2011-01-01

    Since the Viking missions in the mid-1970s, traditional culture-based methods have been used for microbial enumeration by various NASA programs. Viable microbes are of particular concern for spacecraft cleanliness, for forward contamination of extraterrestrial bodies (proliferation of microbes), and for crew health/safety (viable pathogenic microbes). However, a "true" estimation of viable microbial population and differentiation from their dead cells using the most sensitive molecular methods is a challenge, because of the stability of DNA from dead cells. The goal of this research is to evaluate a rapid and sensitive microbial detection concept that will selectively estimate viable microbes. Nucleic acid amplification approaches such as the polymerase chain reaction (PCR) have shown promise for reducing time to detection for a wide range of applications. The proposed method is based on the use of a fluorescent DNA intercalating agent, propidium monoazide (PMA), which can only penetrate the membrane of dead cells. The PMA-quenched reaction mixtures can be screened, where only the DNA from live cells will be available for subsequent PCR reaction and microarray detection, and be identified as part of the viable microbial community. An additional advantage of the proposed rapid method is that it will detect viable microbes and differentiate from dead cells in only a few hours, as opposed to less comprehensive culture-based assays, which take days to complete. This novel combination approach is called the PMA-Microarray method. DNA intercalating agents such as PMA have previously been used to selectively distinguish between viable and dead bacterial cells. Once in the cell, the dye intercalates with the DNA and, upon photolysis under visible light, produces stable DNA adducts. DNA cross-linked in this way is unavailable for PCR. Environmental samples suspected of containing a mixture of live and dead microbial cells/spores will be treated with PMA, and then incubated

  8. Fate of viable but non-culturable Listeria monocytogenes in pig manure microcosms

    Directory of Open Access Journals (Sweden)

    Jeremy eDesneux

    2016-03-01

    Full Text Available The fate of two strains of L. monocytogenes and their ability to become viable but non-culturable (VBNC was investigated in microcosms containing piggery effluents (two raw manures and two biologically treated manures stored for two months at 8°C and 20°C. Levels of L. monocytogenes were estimated using the culture method, qPCR, and propidium monoazide treatment combined with qPCR (qPCRPMA. The chemical composition and the microbial community structure of the manures were also analysed. The strains showed similar decline rates and persisted up to 63 days. At day zero, the percentage of VBNC cells among viable cells was higher in raw manures (81.5-94.8% than in treated manures (67.8-79.2%. The changes in their proportion over time depended on the temperature and on the type of effluent: the biggest increase was observed in treated manures at 20°C and the smallest increase in raw manures at 8°C. The chemical parameters had no influence on the behaviour of the strains, but decrease of the persistence of viable cells was associated with an increase in the microbial richness of the manures. This study demonstrated that storing manure altered the culturability of L. monocytogenes, which rapidly entered the VBNC state, and underlines the importance of including VBNC cells when estimating the persistence of the pathogens in farm effluents.

  9. Glioma Surgical Aspirate: A Viable Source of Tumor Tissue for Experimental Research

    Directory of Open Access Journals (Sweden)

    Perry F. Bartlett

    2013-04-01

    Full Text Available Brain cancer research has been hampered by a paucity of viable clinical tissue of sufficient quality and quantity for experimental research. This has driven researchers to rely heavily on long term cultured cells which no longer represent the cancers from which they were derived. Resection of brain tumors, particularly at the interface between normal and tumorigenic tissue, can be carried out using an ultrasonic surgical aspirator (CUSA that deposits liquid (blood and irrigation fluid and resected tissue into a sterile bottle for disposal. To determine the utility of CUSA-derived glioma tissue for experimental research, we collected 48 CUSA specimen bottles from glioma patients and analyzed both the solid tissue fragments and dissociated tumor cells suspended in the liquid waste fraction. We investigated if these fractions would be useful for analyzing tumor heterogeneity, using IHC and multi-parameter flow cytometry; we also assessed culture generation and orthotopic xenograft potential. Both cell sources proved to be an abundant, highly viable source of live tumor cells for cytometric analysis, animal studies and in-vitro studies. Our findings demonstrate that CUSA tissue represents an abundant viable source to conduct experimental research and to carry out diagnostic analyses by flow cytometry or other molecular diagnostic procedures.

  10. Ergonomic analysis jobs in recovered factories.

    Science.gov (United States)

    Cuenca, Gabriela; Zotta, Gastón

    2012-01-01

    With the advent of the deep economic crisis in Argentina on 2001, the recovery of companies through to the creation of the Cooperatives Working Self-Management or Factories Recovered by its workers was constituted as one of the ways in which the salaried disobeyed the increasing unemployment. When the companies turn into recovered factories they tend to leave of side practices that have been seen like imposed by the previous organization and not understanding them as a primary condition for the execution of his tasks. Safety and ergonomics are two disciplines that are no longer considered relevant to the daily work. Therefore this investigation aims to revalue, undergo semantic to give back to a place in every organization analyzed. This research developed a self-diagnostic tool for working conditions, and the environment, present in the recovered factories.

  11. Mitochondrial respiration in human viable platelets-Methodology and influence of gender, age and storage

    DEFF Research Database (Denmark)

    Sjövall, Fredrik; Ehinger, Johannes K H; Marelsson, Sigurður E

    2013-01-01

    Studying whole cell preparations with intact mitochondria and respiratory complexes has a clear benefit compared to isolated or disrupted mitochondria due to the dynamic interplay between mitochondria and other cellular compartments. Platelet mitochondria have a potential to serve as a source...... of human viable mitochondria when studying mitochondrial physiology and pathogenic mechanisms, as well as for the diagnostics of mitochondrial diseases. The objective of the present study was to perform a detailed evaluation of platelet mitochondrial respiration using high-resolution respirometry. Further...

  12. Recovering core identity, hermeneutical and contextual preaching

    African Journals Online (AJOL)

    2016-07-08

    Jul 8, 2016 ... identity – certainly when it answers the questions 'who are we?' and 'what are we called to be and/or do' – it lacks significantly in its missional identity. Contributing factors are mentioned and remedial action is proposed. Recovering core identity, hermeneutical and contextual preaching. Read online:.

  13. Susceptibility of Some Bacterial Contaminants Recovered from ...

    African Journals Online (AJOL)

    Methods: Nine bacterial isolates recovered from various brands of commercially available cosmetics marketed in Jordan were tested for their susceptibility pattern against two paraben esters and two formaldehyde donors in addition to nine commonly used antibiotics. The biocidal effect for three preservatives was tested at ...

  14. Stress Levels of Recovering Drug Addicts.

    Science.gov (United States)

    LaMon, Brent C.; Alonzo, Anthony

    It appears that chronic drug use may develop as a means of coping in which individuals use self-medication to produce a more desirable state of being. Because drugs are often used to cope with stress, this study examined stress among recovering male drug addicts (N=23) from an urban substance abuse program by administering a self-report inventory…

  15. Applications for Energy Recovering Free Electron Lasers

    Energy Technology Data Exchange (ETDEWEB)

    George Neil

    2007-08-01

    The availability of high-power, high-brilliance sources of tunable photons from energy-recovered Free Electron Lasers is opening up whole new fields of application of accelerators in industry. This talk will review some of the ideas that are already being put into production, and some of the newer ideas that are still under development.

  16. Recovering from Opioid Overdose: Resources for Overdose Survivors & Family Members

    Science.gov (United States)

    SAMHSA Opioid Overdose Prevention TOOLKIT: Recovering From Opioid Overdose – Resources for Overdose Survivors & Family Members TABLE OF CONTENTS Recovering From Opioid Overdose Recovering from Opioid Overdose. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 Resources for Overdose Survivors ...

  17. Detection of viable Salmonella in lettuce by propidium monoazide real-time PCR.

    Science.gov (United States)

    Liang, Ningjian; Dong, Jin; Luo, Laixin; Li, Yong

    2011-05-01

    Contamination of lettuce by Salmonella has caused serious public health problems. Polymerase chain reaction (PCR) allows rapid detection of pathogenic bacteria in food, but it is inaccurate as it might amplify DNA from dead target cells as well. This study aimed to investigate the stability of DNA of dead Salmonella cells in lettuce and to develop an approach to detecting viable Salmonella in lettuce. Salmonella-free lettuce was inoculated with heat-killed Salmonella Typhimurium cells and stored at 4 °C. Bacterial DNA extracted from the sample was amplified by real-time PCR targeting the invA gene. Our results indicate that DNA from the dead cells remained stable in lettuce for at least 8 d. To overcome this limitation, propidium monoazide (PMA), a dye that can selectively penetrate dead bacterial cells and cross-link their DNA upon light exposure, was combined with real-time PCR. Lettuce samples inoculated with different levels of dead or viable S. Typhimurium cells were treated or untreated with PMA before DNA extraction. Real-time PCR suggests that PMA treatment effectively prevented PCR amplification from as high as 10(8) CFU/g dead S. Typhimurium cells in lettuce. The PMA real-time PCR assay could detect viable Salmonella at as low as 10(2) CFU/mL in pure culture and 10(3) CFU/g in lettuce. With 12-h enrichment, S. Typhimurium of 10(1) CFU/g in lettuce was detectable. In conclusion, the PMA real-time PCR assay provides an alternative to real-time PCR assay for accurate detection of Salmonella in food. © 2011 Institute of Food Technologists®

  18. An advanced PCR method for the specific detection of viable total coliform bacteria in pasteurized milk.

    Science.gov (United States)

    Soejima, Takashi; Minami, Jun-ichi; Yaeshima, Tomoko; Iwatsuki, Keiji

    2012-07-01

    Pasteurized milk is a complex food that contains various inhibitors of polymerase chain reaction (PCR) and may contain a large number of dead bacteria, depending on the milking conditions and environment. Ethidium monoazide bromide (EMA)-PCR is occasionally used to distinguish between viable and dead bacteria in foods other than pasteurized milk. EMA is a DNA-intercalating dye that selectively permeates the compromised cell membranes of dead bacteria and cleaves DNA. Usually, EMA-PCR techniques reduce the detection of dead bacteria by up to 3.5 logs compared with techniques that do not use EMA. However, this difference may still be insufficient to suppress the amplification of DNA from dead Gram-negative bacteria (e.g., total coliform bacteria) if they are present in pasteurized milk in large numbers. Thus, false positives may result. We developed a new method that uses real-time PCR targeting of a long DNA template (16S-23S rRNA gene, principally 2,451 bp) following EMA treatment to completely suppress the amplification of DNA of up to 7 logs (10(7) cells) of dead total coliforms. Furthermore, we found that a low dose of proteinase K (25 U/ml) removed PCR inhibitors and simultaneously increased the signal from viable coliform bacteria. In conclusion, our simple protocol specifically detects viable total coliforms in pasteurized milk at an initial count of ≥1 colony forming unit (CFU)/2.22 ml within 7.5 h of total testing time. This detection limit for viable cells complies with the requirements for the analysis of total coliforms in pasteurized milk set by the Japanese Sanitation Act (which specifies <1 CFU/2.22 ml).

  19. Microfiltration of enzyme treated egg whites for accelerated detection of viable Salmonella.

    Science.gov (United States)

    Ku, Seockmo; Ximenes, Eduardo; Kreke, Thomas; Foster, Kirk; Deering, Amanda J; Ladisch, Michael R

    2016-11-01

    We report detection of egg white within 7 h by concentrating the bacteria using microfiltration through 0.2-μm cutoff polyethersulfone hollow fiber membranes. A combination of enzyme treatment, controlled cross-flow on both sides of the hollow fibers, and media selection were key to controlling membrane fouling so that rapid concentration and the subsequent detection of low numbers of microbial cells were achieved. We leveraged the protective effect of egg white proteins and peptone so that the proteolytic enzymes did not attack the living cells while hydrolyzing the egg white proteins responsible for fouling. The molecular weight of egg white proteins was reduced from about 70 kDa to 15 kDa during hydrolysis. This enabled a 50-fold concentration of the cells when a volume of 525 mL of peptone and egg white, containing 13 CFU of Salmonella, was decreased to a 10 mL volume in 50 min. A 10-min microcentrifugation step further concentrated the viable Salmonella cells by 10×. The final cell recovery exceeded 100%, indicating that microbial growth occurred during the 3-h processing time. The experiments leading to rapid concentration, recovery, and detection provided further insights on the nature of membrane fouling enabling fouling effects to be mitigated. Unlike most membrane processes where protein recovery is the goal, recovery of viable microorganisms for pathogen detection is the key measure of success, with modification of cell-free proteins being both acceptable and required to achieve rapid microfiltration of viable microorganisms. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1464-1471, 2016. © 2016 American Institute of Chemical Engineers.

  20. A direct viable count method for the enumeration of attached bacteria and assessment of biofilm disinfection

    Science.gov (United States)

    Yu, F. P.; Pyle, B. H.; McFeters, G. A.

    1993-01-01

    This report describes the adaptation of an in situ direct viable count (in situ DVC) method in biofilm disinfection studies. The results obtained with this technique were compared to two other enumeration methods, the plate count (PC) and conventional direct viable count (c-DVC). An environmental isolate (Klebsiella pneumoniae Kp1) was used to form biofilms on stainless steel coupons in a stirred batch reactor. The in situ DVC method was applied to directly assess the viability of bacteria in biofilms without disturbing the integrity of the interfacial community. As additional advantages, the results were observed after 4 h instead of the 24 h incubation time required for colony formation and total cell numbers that remained on the substratum were enumerated. Chlorine and monochloramine were used to determine the susceptibilities of attached and planktonic bacteria to disinfection treatment using this novel analytical approach. The planktonic cells in the reactor showed no significant change in susceptibility to disinfectants during the period of biofilm formation. In addition, the attached cells did not reveal any more resistance to disinfection than planktonic cells. The disinfection studies of young biofilms indicated that 0.25 mg/l free chlorine (at pH 7.2) and 1 mg/l monochloramine (at pH 9.0) have comparable disinfection efficiencies at 25 degrees C. Although being a weaker disinfectant, monochloramine was more effective in removing attached bacteria from the substratum than free chlorine. The in situ DVC method always showed at least one log higher viable cell densities than the PC method, suggesting that the in situ DVC method is more efficient in the enumeration of biofilm bacteria. The results also indicated that the in situ DVC method can provide more accurate information regarding the cell numbers and viability of bacteria within biofilms following disinfection.

  1. Eukaryotic Cell Invasion does not correlate to flaA SVR Sequence Type based on a Library of Genetically Diverse Campylobacter jejuni Isolates Originally Recovered from A Variety of Sources in Iceland

    Science.gov (United States)

    Introduction: Campylobacter spp. are considered to be a leading bacterial etiologic agent of acute food-borne gastroenteritis among human populations. Epithelial cell invasion is hypothesized to be necessary for human infection and cell invasion assays have been utilized to demonstrate that distinc...

  2. The search for viable local government system in Nigeria: an ...

    African Journals Online (AJOL)

    The history of the Nigerian local government system has been one long episode of trails and errors aimed at achieving viable local government institution without much success. Local government in the country began its long series of reforms from the colonial period when the colonial government attempted to ...

  3. Comment: Towards a Viable Local Government Structure in Nigeria ...

    African Journals Online (AJOL)

    Local governments are principally established for development at the grassroots and they must be structured in a manner that makes them viable and capable of achieving this purpose. The objective of this comment is to appraise the current local government structure under the Nigerian constitutional framework with a view ...

  4. Cultivation and multiplication of viable axenic Trypanosoma vivax in ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-09-01

    Sep 1, 2009 ... Cultivation and multiplication of viable axenic. Trypanosoma vivax in vitro and in vivo. O. A. Idowu, A. B. Idowu, C. F. Mafiana and S. O. Sam-Wobo*. Parasitology Laboratory, Department of Biological Sciences, University of Agriculture, Abeokuta, Nigeria. Accepted 13 April, 2006. Trypanosoma vivax was ...

  5. Recovering of images degraded by atmosphere

    Science.gov (United States)

    Lin, Guang; Feng, Huajun; Xu, Zhihai; Li, Qi; Chen, Yueting

    2017-08-01

    Remote sensing images are seriously degraded by multiple scattering and bad weather. Through the analysis of the radiative transfer procedure in atmosphere, an image atmospheric degradation model considering the influence of atmospheric absorption multiple scattering and non-uniform distribution is proposed in this paper. Based on the proposed model, a novel recovering method is presented to eliminate atmospheric degradation. Mean-shift image segmentation and block-wise deconvolution are used to reduce time cost, retaining a good result. The recovering results indicate that the proposed method can significantly remove atmospheric degradation and effectively improve contrast compared with other removal methods. The results also illustrate that our method is suitable for various degraded remote sensing, including images with large field of view (FOV), images taken in side-glance situations, image degraded by atmospheric non-uniform distribution and images with various forms of clouds.

  6. Rescuing and recovering lost or endangered data

    Directory of Open Access Journals (Sweden)

    Elizabeth Griffin

    2006-01-01

    Full Text Available This paper summarizes the vital importance to the scientific community of rescuing historic scientific data, presently in various informal, non-digital formats, from likely oblivion and making them accessible digitally for trend analyses. It proposes models whereby historic biodiversity and astronomical data can be recovered as Canadian initiatives, in the hope of stimulating further discussion of such simple yet essential rescue missions in the context of Canadian scientific research.

  7. Recovering hydrocarbons with surfactants from lignin

    Energy Technology Data Exchange (ETDEWEB)

    Naae, D.G.; Whittington, L.E.; Ledoux, W.A.; Debons, F.E.

    1988-11-29

    This patent describes a method of recovering hydrocarbons from an underground hydrocarbon formation penetrated by at least one injection well and at least one production well, which comprises: injecting into the formation through an injection well a surfactant slug comprising about 0.1% to about 10% by weight of surfactants produced from lignin, the surfactants produced by placing lignin in contact with water, converting the lignin into low molecular weight lignin phenols by reducing the lignin in the presence of a reducing agent of carbon monoxide or hydrogen creating a reduction reaction mixture comprising oil soluble lignin phenols, the reduction occurring at a temperature greater than about 200/sup 0/C and a pressure greater than about 100 psi, recovering the oil soluble lignin phenols from the reduction mixture, and converting the lignin phenols into lignin surfactants by a reaction selected from the group consisting of alkoxylation, sulfonation, sulfation, aklylation, sulfomethylation, and alkoxysulfation; injecting into the formation through the injection well a drive fluid to push the surfactant slug towards a production well; and recovering hydrocarbons at the production well.

  8. Method for recovering metals from waste

    Science.gov (United States)

    Wicks, G.G.; Clark, D.E.; Schulz, R.L.

    1998-12-01

    A method is described for recovering metals from metals-containing wastes, and vitrifying the remainder of the wastes for disposal. Metals-containing wastes such as circuit boards, cathode ray tubes, vacuum tubes, transistors and so forth, are broken up and placed in a suitable container. The container is heated by microwaves to a first temperature in the range of approximately 300--800 C to combust organic materials in the waste, then heated further to a second temperature in the range of approximately 1,000--1,550 C at which temperature glass formers present in the waste will cause it to melt and vitrify. Low-melting-point metals such as tin and aluminum can be recovered after organics combustion is substantially complete. Metals with higher melting points, such as gold, silver and copper, can be recovered from the solidified product or separated from the waste at their respective melting points. Network former-containing materials can be added at the start of the process to assist vitrification. 2 figs.

  9. Molecular approaches for viable bacterial population and transcriptional analyses in a rodent model of dental caries.

    Science.gov (United States)

    Klein, M I; Scott-Anne, K M; Gregoire, S; Rosalen, P L; Koo, H

    2012-10-01

    Culturing methods are the primary approach for microbiological analysis of plaque biofilms in rodent models of dental caries. In this study, we developed strategies for the isolation of DNA and RNA from plaque biofilms formed in vivo to analyse the viable bacterial population and gene expression. Plaque biofilm samples from rats were treated with propidium monoazide to isolate DNA from viable cells, and the purified DNA was used to quantify total bacteria and the Streptococcus mutans population via quantitative polymerase chain reaction (qPCR) and specific primers; the same samples were also analysed by counting colony-forming units (CFU). In parallel, RNA was isolated from plaque-biofilm samples (from the same animals) and used for transcriptional analyses via reverse transcription-qPCR. The viable populations of both S. mutans and total bacteria assessed by qPCR were positively correlated with the CFU data (P  0.8). However, the qPCR data showed higher bacterial cell counts, particularly for total bacteria (vs. CFU). Moreover, S. mutans proportion in the plaque biofilm determined by qPCR analysis showed strong correlation with incidence of smooth-surface caries (P = 0.0022, r = 0.71). The purified RNAs presented high RNA integrity numbers (> 7), which allowed measurement of the expression of genes that are critical for S. mutans virulence (e.g. gtfB and gtfC). Our data show that the viable microbial population and the gene expression can be analysed simultaneously, providing a global assessment of the infectious aspect of dental caries. Our approach could enhance the value of the current rodent model in further understanding the pathophysiology of this disease and facilitating the exploration of novel anti-caries therapies. © 2012 John Wiley & Sons A/S.

  10. Concurrent detection of other respiratory viruses in children shedding viable human respiratory syncytial virus.

    Science.gov (United States)

    Gagliardi, T B; Paula, F E; Iwamoto, M A; Proença-Modena, J L; Santos, A E; Camara, A A; Cervi, M C; Cintra, O A L; Arruda, E

    2013-10-01

    Human respiratory syncytial virus (HRSV) is an important cause of respiratory disease. The majority of studies addressing the importance of virus co-infections to the HRSV-disease have been based on the detection of HRSV by RT-PCR, which may not distinguish current replication from prolonged shedding of remnant RNA from previous HRSV infections. To assess whether co-detections of other common respiratory viruses are associated with increased severity of HRSV illnesses from patients who were shedding viable-HRSV, nasopharyngeal aspirates from children younger than 5 years who sought medical care for respiratory infections in Ribeirão Preto (Brazil) were tested for HRSV by immunofluorescence, RT-PCR and virus isolation in cell culture. All samples with viable-HRSV were tested further by PCR for other respiratory viruses. HRSV-disease severity was assessed by a clinical score scale. A total of 266 samples from 247 children were collected and 111 (42%) were HRSV-positive. HRSV was isolated from 70 (63%), and 52 (74%) of them were positive for at least one additional virus. HRSV-positive diseases were more severe than HRSV-negative ones, but there was no difference in disease severity between patients with viable-HRSV and those HRSV-positives by RT-PCR. Co-detection of other viruses did not correlate with increased disease severity. HRSV isolation in cell culture does not seem to be superior to RT-PCR to distinguish infections associated with HRSV replication in studies of clinical impact of HRSV. A high rate of co-detection of other respiratory viruses was found in samples with viable-HRSV, but this was not associated with more severe HRSV infection. Copyright © 2013 Wiley Periodicals, Inc.

  11. Selective detection of viable seed-borne Acidovorax citrulli by real-time PCR with propidium monoazide

    Science.gov (United States)

    Tian, Qian; Feng, Jian-jun; Hu, Jie; Zhao, Wen-jun

    2016-01-01

    In recent years, use of the DNA-intercalating dye propidium monoazide (PMA) in real-time PCR has been reported as a novel method to detect viable bacteria in different types of samples, such as food, environmental, and microbiological samples. In this study, viable cells of Acidovorax citrulli, the causal agent of bacterial seedling blight and fruit blotch, were selectively detected and differentiated from dead cells by real-time fluorescent polymerase chain reaction amplification after the bacterial solution was treated with the DNA-binding dye PMA. The primers and TaqMan probe were based on the A. citrulli genome (Aave_1909, Gene ID: 4669443) and were highly specific for A. citrulli. The detection threshold of this assay was 103 colony-forming units per mL (CFU/mL) in pure cell suspensions containing viable and dead cells and infected watermelon seeds. Application of this assay enables the selective detection of viable cells of A. citrulli and facilitates monitoring of the pathogen in watermelon and melon seeds. PMID:27739469

  12. Biofilms in Full-Scale Drinking Water Ozone Contactors Contribute Viable Bacteria to Ozonated Water.

    Science.gov (United States)

    Kotlarz, Nadine; Rockey, Nicole; Olson, Terese M; Haig, Sarah-Jane; Sanford, Larry; LiPuma, John J; Raskin, Lutgarde

    2018-02-13

    Concentrations of viable microbial cells were monitored using culture-based and culture-independent methods across multichamber ozone contactors in a full-scale drinking water treatment plant. Membrane-intact and culturable cell concentrations in ozone contactor effluents ranged from 1200 to 3750 cells/mL and from 200 to 3850 colony forming units/mL, respectively. Viable cell concentrations decreased significantly in the first ozone contact chamber, but rose, even as ozone exposure increased, in subsequent chambers. Our results implicate microbial detachment from biofilms on contactor surfaces, and from biomass present within lime softening sediments in a hydraulic dead zone, as a possible reason for increasing cell concentrations in water samples from sequential ozone chambers. Biofilm community structures on baffle walls upstream and downstream from the dead zone were significantly different from each other (p = 0.017). The biofilms downstream of the dead zone contained a significantly (p = 0.036) higher relative abundance of bacteria of the genera Mycobacterium and Legionella than the upstream biofilms. These results have important implications as the effluent from ozone contactors is often treated further in biologically active filters and bacteria in ozonated water continuously seed filter microbial communities.

  13. Survival Strategy of Erwinia amylovora against Copper: Induction of the Viable-but-Nonculturable State

    Science.gov (United States)

    Ordax, Mónica; Marco-Noales, Ester; López, María M.; Biosca, Elena G.

    2006-01-01

    Copper compounds, widely used to control plant-pathogenic bacteria, have traditionally been employed against fire blight, caused by Erwinia amylovora. However, recent studies have shown that some phytopathogenic bacteria enter into the viable-but-nonculturable (VBNC) state in the presence of copper. To determine whether copper kills E. amylovora or induces the VBNC state, a mineral medium without copper or supplemented with 0.005, 0.01, or 0.05 mM Cu2+ was inoculated with 107 CFU/ml of this bacterium and monitored over 9 months. Total and viable cell counts were determined by epifluorescence microscopy using the LIVE/DEAD kit and by flow cytometry with 5-cyano-2,3-ditolyl tetrazolium chloride and SYTO 13. Culturable cells were counted on King's B nonselective solid medium. Changes in the bacterial morphology in the presence of copper were observed by scanning electron microscopy. E. amylovora entered into the VBNC state at all three copper concentrations assayed, much faster when the copper concentration increased. The addition of different agents which complex copper allowed the resuscitation (restoration of culturability) of copper-induced VBNC cells. Finally, copper-induced VBNC cells were virulent only for the first 5 days, while resuscitated cells always regained their pathogenicity on immature fruits over 9 months. These results have shown, for the first time, the induction of the VBNC state in E. amylovora as a survival strategy against copper. PMID:16672494

  14. Determination of viable wine yeast using DNA binding dyes and quantitative PCR.

    Science.gov (United States)

    Andorrà, Imma; Esteve-Zarzoso, Braulio; Guillamón, José M; Mas, Albert

    2010-12-15

    The detection and quantification of wine yeast can be misleading due to under or overestimation of these microorganisms. Underestimation may be caused by variable growing rates of different microorganisms in culture media or the presence of viable but non-cultivable microorganisms. Overestimation may be caused by the lack of discrimination between live and dead microorganisms if quantitative PCR is used to quantify with DNA as the template. However, culture-independent methods that use dyes have been described to remove the DNA from dead cells and then quantify the live microorganisms. Two dyes have been studied in this paper: ethidium monoazide bromide (EMA) and propidium monoazide bromide (PMA). The technique was applied to grape must fermentation and ageing wines. Both dyes presented similar results on yeast monitoring. Membrane cell recovery was necessary when yeasts were originated from ethanol-containing media. When applied to grape must fermentation, differences of up to 1 log unit were seen between the QPCR estimation with or without the dye during the stationary phase. In ageing wines, good agreement was found between plating techniques and QPCR. Most of the viable cells were also culturable and no differences were observed with the methods, except for Zygosaccharomyces bailii and Dekkera bruxellensis where much higher counts were occasionally detected by QPCR. The presence of excess dead cells did not interfere with the quantification of live cells with either of the dyes. Copyright © 2010 Elsevier B.V. All rights reserved.

  15. Viable group A streptococci in macrophages during acute soft tissue infection.

    Directory of Open Access Journals (Sweden)

    Pontus Thulin

    2006-03-01

    Full Text Available Group A streptococcal severe soft tissue infections, such as necrotizing fasciitis, are rapidly progressive infections associated with high mortality. Group A streptococcus is typically considered an extracellular pathogen, but has been shown to reside intracellularly in host cells.We characterized in vivo interactions between group A streptococci (GAS and cells involved in innate immune responses, using human biopsies (n = 70 collected from 17 patients with soft tissue infections. Immunostaining and in situ image analysis revealed high amounts of bacteria in the biopsies, even in those collected after prolonged antibiotic therapy. Viability of the streptococci was assessed by use of a bacterial viability stain, which demonstrated viable bacteria in 74% of the biopsies. GAS were present both extracellularly and intracellularly within phagocytic cells, primarily within macrophages. Intracellular GAS were predominantly noted in biopsies from newly involved tissue characterized by lower inflammation and bacterial load, whereas purely extracellular GAS or a combination of intra- and extracellular GAS dominated in severely inflamed tissue. The latter tissue was also associated with a significantly increased amount of the cysteine protease streptococcal pyrogenic exotoxin SpeB. In vitro studies confirmed that macrophages serve as reservoirs for viable GAS, and infection with a speB-deletion mutant produced significantly lower frequencies of cells with viable GAS following infection as compared to the wild-type bacteria.This is the first study to demonstrate that GAS survive intracellularly in macrophages during acute invasive infections. This intracellular presence may have evolved as a mechanism to avoid antibiotic eradication, which may explain our finding that high bacterial load is present even in tissue collected after prolonged intravenous antibiotic therapy. This new insight into the pathogenesis of streptococcal soft tissue infections

  16. Viable Group A Streptococci in Macrophages during Acute Soft Tissue Infection.

    Directory of Open Access Journals (Sweden)

    2006-01-01

    Full Text Available BACKGROUND: Group A streptococcal severe soft tissue infections, such as necrotizing fasciitis, are rapidly progressive infections associated with high mortality. Group A streptococcus is typically considered an extracellular pathogen, but has been shown to reside intracellularly in host cells. METHODS AND FINDINGS: We characterized in vivo interactions between group A streptococci (GAS and cells involved in innate immune responses, using human biopsies (n = 70 collected from 17 patients with soft tissue infections. Immunostaining and in situ image analysis revealed high amounts of bacteria in the biopsies, even in those collected after prolonged antibiotic therapy. Viability of the streptococci was assessed by use of a bacterial viability stain, which demonstrated viable bacteria in 74% of the biopsies. GAS were present both extracellularly and intracellularly within phagocytic cells, primarily within macrophages. Intracellular GAS were predominantly noted in biopsies from newly involved tissue characterized by lower inflammation and bacterial load, whereas purely extracellular GAS or a combination of intra- and extracellular GAS dominated in severely inflamed tissue. The latter tissue was also associated with a significantly increased amount of the cysteine protease streptococcal pyrogenic exotoxin SpeB. In vitro studies confirmed that macrophages serve as reservoirs for viable GAS, and infection with a speB-deletion mutant produced significantly lower frequencies of cells with viable GAS following infection as compared to the wild-type bacteria. CONCLUSIONS: This is the first study to demonstrate that GAS survive intracellularly in macrophages during acute invasive infections. This intracellular presence may have evolved as a mechanism to avoid antibiotic eradication, which may explain our finding that high bacterial load is present even in tissue collected after prolonged intravenous antibiotic therapy. This new insight into the pathogenesis

  17. Acupuntura un tratamiento viable para las adicciones en Colombia

    Directory of Open Access Journals (Sweden)

    Hernán López Seuscún

    2013-07-01

    Los tratamientos con auriculoterapia, como el protocolo NADA (National Acupuncture Detoxification Association, son los métodos más usados para las adicciones en el mundo, y aunque no se ha logrado evidenciar su efectividad, por su costo, facilidad y el poco riesgo de efectos adversos se hace viable en un país con pocos recursos económicos como Colombia.

  18. Academic Pediatric Dentistry is a Rewarding, Financially Viable Career Path.

    Science.gov (United States)

    Townsend, Janice A; Chi, Donald L

    2017-09-15

    Newly graduated pediatric dentists have unprecedented levels of debt. High levels of student debt may be perceived as an obstacle to pursue an academic career. However, opportunities exist through faculty compensation models and loan repayment programs that make an academic career financially viable. The purpose of this paper is to outline the benefits of a career in academic dentistry and provide examples of young pediatric dentistry faculty members who have been able to manage student debt while pursuing meaningful and rewarding careers.

  19. How Can We Prevent Violence Becoming a Viable Political Strategy?

    OpenAIRE

    Patricia Justino

    2009-01-01

    A basic issue that conflict analysis investigates is how non-peaceful ways of living and governing become viable political strategies. Macro-level studies provide some important insights but micro-level analysis is vital to understand the mechanisms that make violence possible. This briefing outlines some preliminary findings in this respect from MICROCON, a major research programme analysing violent conflict at the micro level. It also discusses their implications for policies aimed at preve...

  20. Mice carrying a complete deletion of the talin2 coding sequence are viable and fertile

    Energy Technology Data Exchange (ETDEWEB)

    Debrand, Emmanuel; Conti, Francesco J.; Bate, Neil; Spence, Lorraine; Mazzeo, Daniela; Pritchard, Catrin A.; Monkley, Susan J. [Department of Biochemistry, University of Leicester, Lancaster Road, Leicester LE1 9HN (United Kingdom); Critchley, David R., E-mail: drc@le.ac.uk [Department of Biochemistry, University of Leicester, Lancaster Road, Leicester LE1 9HN (United Kingdom)

    2012-09-21

    Highlights: Black-Right-Pointing-Pointer Mice lacking talin2 are viable and fertile with only a mildly dystrophic phenotype. Black-Right-Pointing-Pointer Talin2 null fibroblasts show no major defects in proliferation, adhesion or migration. Black-Right-Pointing-Pointer Maintaining a colony of talin2 null mice is difficult indicating an underlying defect. -- Abstract: Mice homozygous for several Tln2 gene targeted alleles are viable and fertile. Here we show that although the expression of talin2 protein is drastically reduced in muscle from these mice, other tissues continue to express talin2 albeit at reduced levels. We therefore generated a Tln2 allele lacking the entire coding sequence (Tln2{sup cd}). Tln2{sup cd/cd} mice were viable and fertile, and the genotypes of Tln2{sup cd/+} intercrosses were at the expected Mendelian ratio. Tln2{sup cd/cd} mice showed no major difference in body mass or the weight of the major organs compared to wild-type, although they displayed a mildly dystrophic phenotype. Moreover, Tln2{sup cd/cd} mouse embryo fibroblasts showed no obvious defects in cell adhesion, migration or proliferation. However, the number of Tln2{sup cd/cd} pups surviving to adulthood was variable suggesting that such mice have an underlying defect.

  1. Recovering an obstacle using integral equations

    KAUST Repository

    Rundell, William

    2009-05-01

    We consider the inverse problem of recovering the shape, location and surface properties of an object where the surrounding medium is both conductive and homogeneous and we measure Cauchy data on an accessible part of the exterior boundary. It is assumed that the physical situation is modelled by harmonic functions and the boundary condition on the obstacle is one of Dirichlet type. The purpose of this paper is to answer some of the questions raised in a recent paper that introduced a nonlinear integral equation approach for the solution of this type of problem.

  2. Five years post whiplash injury: Symptoms and psychological factors in recovered versus non-recovered

    Directory of Open Access Journals (Sweden)

    Stålnacke Britt-Marie

    2010-07-01

    Full Text Available Abstract Background Few studies have focused on the differences between persons who are recovered after whiplash injury and those who suffer from persistent disability. The primary aim of this study was therefore to examine differences in symptoms, psychological factors and life satisfaction between subjects classified as recovered and those with persistent disability five years after whiplash injury based on the Neck Disability Index (NDI. Methods A set of questionnaires was answered by 158 persons (75 men, 83 women to assess disability (NDI, pain intensity (VAS, whiplash-related symptoms (Rivermead Post-Concussion Symptoms Questionnaire, RPQ, post-traumatic stress (Impact of Event Scale, IES, depression (Beck's depression inventory, BDI and life satisfaction (LiSat-11. The participants were divided into three groups based on the results of the NDI: recovered (34.8%, mild disability (37.3% and moderate/severe disability (27.3%. Results The moderate/severe group reported significantly higher VAS, BDI and IES scores and lower level of physical health and psychological health compared to the mild and the recovered groups. Less significant differences were reported between the mild and the recovered groups. Conclusions The group with the highest disability score reported most health problems with pain, symptoms, depression, post-traumatic stress and decreased life satisfaction. These findings indicate that classifying these subjects into subgroups based on disability levels makes it possible to optimize the management and treatment after whiplash injury.

  3. Recovering energy and materials from hazardous waste

    Energy Technology Data Exchange (ETDEWEB)

    Anon.

    2003-12-01

    The tannery industry faces growing environmental concerns because of the high hazardous metal content of its process waste. The formation, during the tanning process, of the highly toxic hexavalent chromium precludes the use of conventional thermal incineration processes. Borge Tannery in Norway, which processes 600 cattle hides per day, has solved the problem by using new PyroArc technology. The PyroArc waste processing plant can treat all of the tannery's production wastes, transforming them into useful products such as fuel gas and re-usable metal. The fuel gas consists mainly of carbon monoxide, hydrogen and nitrogen, and has a calorific value of about 4 MJ/Nm{sub 3}. About 65-70% of the energy content of the source material (waste or biomass) is recovered in the gas, and this is used to produce steam and/or electricity in a gas engine with a capacity of 580 kW. A further 20-25% of the initial energy content is recovered as heat or low-pressure steam. The plant is designed to be self-sufficient in energy (1.5 MW) and to meet the tannery's maximum requirements for hot water and steam. (UK)

  4. When Will the Antarctic Ozone Hole Recover?

    Science.gov (United States)

    Newman, Paul A.

    2006-01-01

    The Antarctic ozone hole develops each year and culminates by early spring (late September - early October). Antarctic ozone values have been monitored since 1979 using satellite observations from the TOMS instrument. The severity of the hole has been assessed from TOMS using the minimum total ozone value from the October monthly mean (depth of the hole) and by calculating the average area coverage during this September-October period. Ozone is mainly destroyed by halogen (chlorine and bromine) catalytic cycles, and these losses are modulated by temperature variations in the collar of the polar lower stratospheric vortex. In this talk, I will show the relationships of halogens and temperature to both the size and depth of the hole. Because atmospheric halogen levels are responding to international agreements that limit or phase out production, the amount of halogens in the stratosphere should decrease over the next few decades. Using projections of halogen levels combined with age-of-air estimates, we find that the ozone hole is recovering at an extremely slow rate and that large ozone holes will regularly recur over the next 2 decades. The ozone hole will begin to show first signs of recovery in about 2023, and the hole will fully recover to pre-1980 levels in approximately 2070. This 2070 recovery is 20 years later than recent projections. I will also discuss current assessments of mid-latitude ozone recovery.

  5. When will the Antarctic Ozone Hole Recover?

    Science.gov (United States)

    Newman, Paul A.; Nash, Eric R.; Kawa, S. Randolph; Montzka, Steve

    2006-01-01

    The Antarctic ozone hole develops each year and culminates by early Spring. Antarctic ozone values have been monitored since 1979 using satellite observations from the .TOMS instrument. The severity of the hole has been assessed from TOMS using the minimum total ozone value from the October monthly mean (depth of the hole) and by calculating the average size during the September-October period. Ozone is mainly destroyed by halogen catalytic cycles, and these losses are modulated by temperature variations in the collar of the polar lower stratospheric vortex. In this presentation, we show the relationships of halogens and temperature to, both the size and depth of the hole. Because atmospheric halogen levels are responding to international agreements that limit or phase out production, the amount of halogens in the stratosphere should decrease over the next few decades. Using projections of halogen levels combined with age-of-air estimates, we find that the ozone hole is recovering at an extremely slow rate and that large ozone holes will regularly recur over the next 2 decades. The ozone hole will begin to show first signs of recovery in about 2023, and the hole will fully recover to pre-1980 levels in approximately 2070. This 2070 recovery is 20 years later than recent projections.

  6. Sorption and precipitation of Mn2+ by viable and autoclaved Shewanella putrefaciens: Effect of contact time

    KAUST Repository

    Chubar, Natalia

    2013-01-01

    The sorption of Mn(II) by viable and inactivated cells of Shewanella putrefaciens, a non-pathogenic, facultative anaerobic, gram-negative bacterium characterised as a Mn(IV) and Fe(III) reducer, was studied under aerobic conditions, as a function of pH, bacterial density and metal loading. During a short contact time (3-24h), the adsorptive behaviour of live and dead bacteria toward Mn(II) was sufficiently similar, an observation that was reflected in the studies on adsorption kinetics at various metal loadings, effects of pH, bacteria density, isotherms and drifting of pH during adsorption. Continuing the experiment for an additional 2-30days demonstrated that the Mn(II) sorption by suspensions of viable and autoclaved cells differed significantly from one another. The sorption to dead cells was characterised by a rapid equilibration and was described by an isotherm. In contrast, the sorption (uptake) to live bacteria exhibited a complex time-dependent uptake. This uptake began as adsorption and ion exchange processes followed by bioprecipitation, and it was accompanied by the formation of polymeric sugars (EPS) and the release of dissolved organic substances. FTIR, EXAFS/XANES and XPS demonstrated that manganese(II) phosphate was the main precipitate formed in 125ml batches, which is the first evidence of the ability of microbes to synthesise manganese phosphates. XPS and XANES spectra did not detect Mn(II) oxidation. Although the release of protein-like compounds by the viable bacteria increased in the presence of Mn2+ (and, by contrast, the release of carbohydrates did not change), electrochemical analyses did not indicate any aqueous complexation of Mn(II) by the organic ligands. © 2012 Elsevier Ltd.

  7. Liquid biopsy in cancer patients: advances in capturing viable CTCs for functional studies using the EPISPOT assay.

    Science.gov (United States)

    Alix-Panabières, Catherine; Pantel, Klaus

    2015-01-01

    Circulating tumor cells (CTCs) in the blood of cancer patients have received increasing attention as new diagnostic tool enabling 'liquid biopsies'. In contrast to the wealth of descriptive studies demonstrating the clinical relevance of CTCs as biomarkers, the extremely low concentration of CTCs in the peripheral blood of most cancer patients challenges further functional studies. This article discusses the current possibilities to enrich and, in particular, detect viable CTCs with emphasis on the EPithelial ImmunoSPOT technology. This functional assay detects viable CTCs at the single-cell level and has been used on hundreds of patients with different tumor types including epithelial tumors (breast, prostate and colon cancer) and melanomas. Moreover, the article summarizes recent advances in the in vitro and in vivo expansion of CTCs from cancer patients. These functional analyses will contribute to identifying the biological properties of metastatic cells and reveal new therapeutic targets against disseminating cancer cells.

  8. Modulation of the endocannabinoid system in viable and non-viable first trimester pregnancies by pregnancy-related hormones

    Directory of Open Access Journals (Sweden)

    Taylor Anthony H

    2011-11-01

    Full Text Available Abstract Background In early pregnancy, increased plasma levels of the endocannabinoid anandamide (AEA are associated with miscarriage through mechanisms that might affect the developing placenta or maternal decidua. Methods In this study, we compare AEA levels in failed and viable pregnancies with the levels of the trophoblastic hormones (beta-human chorionic gonadotrophin (beta-hCG, progesterone (P4 and (pregnancy-associated placental protein-A (PAPP-A essential for early pregnancy success and relate that to the expression of the cannabinoid receptors and enzymes that modulate AEA levels. Results The median plasma AEA level in non-viable pregnancies (1.48 nM; n = 20 was higher than in viable pregnancies (1.21 nM; n = 25; P = 0.013, as were progesterone and beta-hCG levels (41.0 vs 51.5 ng/mL; P = 0.052 for P4 and 28,650 vs 6,560 mIU/L; P = 0.144 for beta-hCG, respectively, but were not statistically significant. Serum PAPP-A levels in the viable group were approximately 6.8 times lower than those in the non-viable group (1.82 vs 12.25 mg/L; P = 0.071, but again these differences were statistically insignificant. In the spontaneous miscarriage group, significant correlations between P4 and beta-hCG, P4 and PAPP-A and AEA and PAPP-A levels were observed. Simultaneously, immunohistochemical distributions of the two main cannabinoid receptors and the AEA-modifying enzymes, fatty acid amide hydrolase (FAAH and N-acylphosphatidylethanolamine-phospholipase D (NAPE-PLD, changed within both the decidua and trophoblast. Conclusions The association of higher AEA levels with early pregnancy failure and with beta-hCG and PAPP-A, but not with progesterone concentrations suggest that plasma AEA levels and pregnancy failure are linked via a mechanism that may involve trophoblastic beta-hCG, and PAPP-A, but not, progesterone production. Although the trophoblast, decidua and embryo contain receptors for AEA, the main AEA target in early pregnancy failure

  9. Removal of viable bioaerosol particles with a low-efficiency HVAC filter enhanced by continuous emission of unipolar air ions.

    Science.gov (United States)

    Huang, R; Agranovski, I; Pyankov, O; Grinshpun, S

    2008-04-01

    Continuous emission of unipolar ions has been shown to improve the performance of respirators and stationary filters challenged with non-biological particles. In this study, we investigated the ion-induced enhancement effect while challenging a low-efficiency heating, ventilation and air-conditioning (HVAC) filter with viable bacterial cells, bacterial and fungal spores, and viruses. The aerosol concentration was measured in real time. Samples were also collected with a bioaerosol sampler for viable microbial analysis. The removal efficiency of the filter was determined, respectively, with and without an ion emitter. The ionization was found to significantly enhance the filter efficiency in removing viable biological particles from the airflow. For example, when challenged with viable bacteria, the filter efficiency increased as much as four- to fivefold. For viable fungal spores, the ion-induced enhancement improved the efficiency by a factor of approximately 2. When testing with virus-carrying liquid droplets, the original removal efficiency provided by the filter was rather low: 9.09 +/- 4.84%. While the ion emission increased collection about fourfold, the efficiency did not reach 75-100% observed with bacteria and fungi. These findings, together with our previously published results for non-biological particles, demonstrate the feasibility of a new approach for reducing aerosol particles in HVAC systems used for indoor air quality control. Recirculated air in HVAC systems used for indoor air quality control in buildings often contains considerable number of viable bioaerosol particles because of limited efficiency of the filters installed in these systems. In the present study, we investigated - using aerosolized bacterial cells, bacterial and fungal spores, and virus-carrying particles - a novel idea of enhancing the performance of a low-efficiency HVAC filter utilizing continuous emission of unipolar ions in the filter vicinity. The findings described in

  10. Molar Pregnancy with a Co-Existing Viable Fetus

    Directory of Open Access Journals (Sweden)

    Ruya Deveer

    2014-03-01

    Full Text Available     The aim of this study was to report the clinical features, management, and outcome of a case of molar pregnancy with a coexisting viable fetus and to review the literature. In this article, we report a case of pregnancy with diffuse placental molar change and a normal fetus which presented with hyperemesis gravidarum and hyperthyroidism. Genetic amniocentesis showed normal fetal karyotype. A healthy full-term live male infant was delivered by cesarean section. In molar pregnancies with a normal karyotype fetus, with intensive maternal follow-up, continuation of pregnancy can be suggested.

  11. Use of sodium lauroyl sarcosinate (sarkosyl) in viable real-time PCR for enumeration of Escherichia coli.

    Science.gov (United States)

    Wang, Hui; Gill, Colin O; Yang, Xianqin

    2014-03-01

    The cell membranes of inactivated Escherichia coli are not always permeable to propidium monoazide (PMA). This limits the use of PMA real-time PCR (PMA-qPCR) for quantification of DNA from only viable cells for enumeration of E. coli. The aim of this study was to develop PMA-qPCR procedures for E. coli with improved selectivity for viable cells. E. coli inactivated by incubation at 52°C were treated with 12 detergents before PMA treatment, and DNA was quantified by real-time PCR. Treatment with each of the 12 detergents and PMA increased the cycle threshold (Ct) values for heat inactivated E. coli suspensions. The greatest increase, of 10.68 Ct was obtained with sarkosyl. Treatment with sodium deoxycholate (NaDC) increased the Ct value by 8.99 Ct. Treatment with sarkosyl or NaDC of 16 heat treated 5-strain cocktails of verotoxigenic E. coli (VTEC) increased the mean Ct values by 8.15 or 6.82 Ct, respectively. Those mean values were significantly (pnumbers of viable E. coli were 2.24 and 2.47, respectively, with regression coefficient values ≥0.85. The findings show that sarkosyl was more effective than NaDC for dissipation of PMA-barrier properties of membranes of inactivated E. coli cells. Viable E. coli in mixtures of viable E. coli and E. coli inactivated by heat, lactic acid or peroxyacetic acid could be reliably enumerated by sarkosyl PMA-qPCR. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Whole-cell protein profiles are useful for distinguishing enterococcal species recovered from clinical specimens Los perfiles de proteínas totales son útiles para distinguir especies de enterococos recuperados de muestras clínicas

    Directory of Open Access Journals (Sweden)

    R. Massa

    2007-12-01

    Full Text Available Whole-cell protein analysis was performed for differentiating 150 enterococcal isolates to the species level, which had previously been identified by extended phenotypic conventional tests. Whole-cell protein profile (WCPP showed a high degree of similarity within species and comparison between species revealed important differences in band profiles. All Enterococcus faecalis and Enterococcus faecium isolates were properly located into their corresponding species, regardless of their clinical source and susceptibility pattern. Moreover, WCPP allowed relocation of some isolates that had erroneously been identified by the usual conventional scheme (i.e. two atypical arginine-negative E. faecalis isolates. WCPP proved to be a simple method to ascertain the various enterococcal species, especially those other than E. faecalis, and may be a suitable tool for high-complexity or reference clinical laboratories.La comparación del perfil de proteínas totales permitió agrupar 150 aislamientos de enterococos dentro de la especie en la que habían sido ubicados por el esquema convencional de pruebas bioquímicas. Los patrones de proteínas totales, comparados visualmente, se mantuvieron con alto grado de similitud intraespecie y revelaron diferencias notorias en la comparación interespecie. Todos los aislamientos de Enterococcus faecalis y Enterococcus faecium, independientemente de los sitios de aislamiento, cuadro clínico del paciente, biotipo o antibiotipo, fueron fácilmente encuadrados en su especie. Asimismo, el estudio del perfil de proteínas totales de enterococos permitió reubicar taxonómicamente aislamientos que habían sido incorrectamente identificados por los métodos bioquímicos convencionales, como por ejemplo dos aislamientos atípicos de E. faecalis arginina negativos. Dado que la metodología empleada es económica y rápida, la comparación de perfiles de proteínas totales en SDS-PAGE podría ser considerada una herramienta

  13. A Novel Korean Red Ginseng Compound Gintonin Inhibited Inflammation by MAPK and NF-κB Pathways and Recovered the Levels of mir-34a and mir-93 in RAW 264.7 Cells

    Science.gov (United States)

    Saba, Evelyn; Jeon, Bo Ra; Jeong, Da-Hye; Lee, Kija; Goo, Youn-Kyoung; Kwak, Dongmi; Kim, Suk; Roh, Seong-Soo; Kim, Sung Dae; Nah, Seung-Yeol; Rhee, Man Hee

    2015-01-01

    The beneficial health promoting effects of ginseng from vitalizing the body to enhancing long life have been well explored very rapidly in the past few years. Up till now many ginsenosides have been discovered for their marvelous therapeutic effects. However during past three years, a novel ginseng compound has been discovered, called gintonin, that differs from other ginsenosides on the basis of its signal transduction and chemical nature. Gintonin has been widely studied for its anti-Alzheimer's disease activities and other neuropathies. However, its anti-inflammatory activity remained unexplored. In our study we have reported for the first time the anti-inflammatory activity of gintonin on RAW 264.7 cells. We found that gintonin potently suppresses the nitric oxide production without any cytotoxicity at given doses and also efficiently suppressed the levels of proinflammatory cytokines. Moreover, it mediaes its signal transduction via MAPK and NF-κB pathways and revives the levels of mir-34a and mir-93. These findings are valuable for the anti-inflammatory effects of this new compound with particular reference to microRNA involvement in the ginseng family. PMID:26579204

  14. 40 CFR 721.4600 - Recovered metal hydroxide.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Recovered metal hydroxide. 721.4600... Substances § 721.4600 Recovered metal hydroxide. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified generically as a recovered metal hydroxide (PMN P-91-809...

  15. Propidium monoazide combined with real-time quantitative PCR to quantify viable Alternaria spp. contamination in tomato products.

    Science.gov (United States)

    Crespo-Sempere, Ana; Estiarte, Núria; Marín, Sonia; Sanchis, Vicente; Ramos, Antonio J

    2013-08-01

    Alternaria is a common contaminating genus of fungi in fruits, grains, and vegetables that causes severe economic losses to farmers and the food industry. Furthermore, it is claimed that Alternaria spp. are able to produce phytotoxic metabolites, and mycotoxins that are unsafe for human and animal health. DNA amplification techniques are being increasingly applied to detect, identify, and quantify mycotoxigenic fungi in foodstuffs, but the inability of these methods to distinguish between viable and nonviable cells might lead to an overestimation of mycotoxin-producing living cells. A promising technique to overcome this problem is the pre-treatment of samples with nucleic acid intercalating dyes, such as propidium monoazide (PMA), prior to quantitative PCR (qPCR). PMA selectively penetrates cells with a damaged membrane inhibiting DNA amplification during qPCRs. In our study, a primer pair (Alt4-Alt5) to specifically amplify and quantify Alternaria spp. by qPCR was designed. Quantification data of qPCR achieved a detection limit of 10(2)conidia/g of tomato. Here, we have optimized for the first time a DNA amplification-based PMA sample pre-treatment protocol for detecting viable Alternaria spp. cells. Artificially inoculated tomato samples treated with 65μM of PMA, showed a reduction in the signal by almost 7cycles in qPCR between live and heat-killed Alternaria spp. conidia. The tomato matrix had a protective effect on the cells against PMA toxicity, reducing the efficiency to distinguish between viable and nonviable cells. The results reported here indicate that the PMA-qPCR method is a suitable tool for quantifying viable Alternaria cells, which could be useful for estimating potential risks of mycotoxin contamination. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. A ketogenic diet increases succinic dehydrogenase (SDH) activity and recovers age-related decrease in numeric density of SDH-positive mitochondria in cerebellar Purkinje cells of late-adult rats.

    Science.gov (United States)

    Balietti, Marta; Giorgetti, Belinda; Di Stefano, Giuseppina; Casoli, Tiziana; Platano, Daniela; Solazzi, Moreno; Bertoni-Freddari, Carlo; Aicardi, Giorgio; Lattanzio, Fabrizia; Fattoretti, Patrizia

    2010-02-01

    Ketogenic diets (KDs) have been applied in the therapy of paediatric epilepsy for nearly a century. Recently, beneficial results have also been reported on metabolic disorders and neurodegeneration, designating aged individuals as possible recipients. However, KDs efficacy decrease after the suckling period, and very little is known about their impact on the aging brain. In the present study, the effect on the neuronal energetic supply of a KD containing 20% of medium chain triglycerides (MCT) was investigated in Purkinje cells of the cerebellar vermis of late-adult (19-month-old) rats. The animals were fed with the KD for 8 weeks, and succinic dehydrogenase (SDH) activity was cytochemically determined. The following parameters of SDH-positive mitochondria were evaluated by the use of a computer-assisted image analysis system connected to a transmission electron microscope: numeric density (Nv), average volume (V), volume density (Vv), and cytochemical precipitate area/mitochondrial area (R). Young, age-matched, and old animals fed with a standard chow were used as controls. We found significantly higher Nv in MCT-KD-fed rats vs. all the control groups, in young vs. late-adult and old controls, and in late-adult vs. old controls. V and Vv showed no significant differences among the groups. R was significantly higher in MCT-KD-fed rats vs. all the control animals, and in old vs. young and late-adult controls. Present data indicate that the ketogenic treatment counteracted age-related decrease in numeric density of SDH-positive mitochondria, and enhanced their metabolic efficiency. Given the central role of mitochondrial impairment in age-related physio-pathological changes of the brain, these findings may represent a starting point to examine novel potentialities for KDs.

  17. Significance of Viable but Nonculturable Escherichia coli: Induction, Detection, and Control.

    Science.gov (United States)

    Ding, Tian; Suo, Yuanjie; Xiang, Qisen; Zhao, Xihong; Chen, Shiguo; Ye, Xingqian; Liu, Donghong

    2017-03-28

    Diseases caused by foodborne or waterborne pathogens are emerging. Many pathogens can enter into the viable but nonculturable (VBNC) state, which is a survival strategy when exposed to harsh environmental stresses. Pathogens in the VBNC state have the ability to evade conventional microbiological detection methods, posing a significant and potential health risk. Therefore, controlling VBNC bacteria in food processing and the environment is of great importance. As the typical one of the gram-negatives, Escherichia coli (E. coli) is a widespread foodborne and waterborne pathogenic bacterium and is able to enter into a VBNC state in extreme conditions (similar to the other gram-negative bacteria), including inducing factors and resuscitation stimulus. VBNC E. coli has the ability to recover both culturability and pathogenicity, which may bring potential health risk. This review describes the concrete factors (nonthermal treatment, chemical agents, and environmental factors) that induce E. coli into the VBNC state, the condition or stimulus required for resuscitation of VBNC E. coli, and the methods for detecting VBNC E. coli. Furthermore, the mechanism of genes and proteins involved in the VBNC E. coli is also discussed in this review.

  18. Identification of Viable Helicobacter pylori in Drinking Water Supplies by Cultural and Molecular Techniques.

    Science.gov (United States)

    Santiago, Paula; Moreno, Yolanda; Ferrús, M Antonía

    2015-08-01

    Helicobacter pylori is one of the most common causes of chronic bacterial infection in humans, directly related to peptic ulcer and gastric cancer. It has been suggested that H. pylori can be acquired through different transmission routes, including water. In this study, culture and qPCR were used to detect and identify the presence of H. pylori in drinking water. Furthermore, the combined techniques PMA-qPCR and DVC-FISH were applied for detection of viable cells of H. pylori. Among 24 drinking water samples, 16 samples were positive for the presence of H. pylori, but viable cells were only detected in six samples. Characteristic colonies, covered by a mass of bacterial unspecific growth, were observed on selective agar plates from an only sample, after enrichment. The mixed culture was submitted to DVC-FISH and qPCR analysis, followed by sequencing of the amplicons. Molecular techniques confirmed the growth of H. pylori on the agar plate. Our results demonstrate for the first time that H. pylori can survive and be potentially infective in drinking water, showing that water distribution systems could be a potential route for H. pylori transmission. © 2015 John Wiley & Sons Ltd.

  19. Real-time quantification of viable bacteria in liquid medium using infrared thermography

    Science.gov (United States)

    Salaimeh, Ahmad A.; Campion, Jeffrey J.; Gharaibeh, Belal Y.; Evans, Martin E.; Saito, Kozo

    2011-11-01

    Quantifying viable bacteria in liquids is important in environmental, food processing, manufacturing, and medical applications. Since vegetative bacteria generate heat as a result of biochemical reactions associated with cellular functions, thermal sensing techniques, including infrared thermography (IRT), have been used to detect viable cells in biologic samples. We developed a novel method that extends the dynamic range and improves the sensitivity of bacterial quantification by IRT. The approach uses IRT video, thermodynamics laws, and heat transfer mechanisms to directly measure, in real-time, the amount of energy lost as heat from the surface of a liquid sample containing bacteria when the specimen cools to a lower temperature over 2 min. We show that the Energy Content ( EC) of liquid media containing as few as 120 colony-forming units (CFU) of Escherichia coli per ml was significantly higher than that of sterile media ( P method that provides real-time bacterial enumeration over a wide dynamic range without the need for sample concentration, modification, or destruction. The approach could be adapted to quantify other living cells in a liquid milieu and has the potential for automation and high throughput.

  20. Allogeneic human dermal fibroblasts are viable in peripheral blood mononuclear co-culture

    Directory of Open Access Journals (Sweden)

    Restu Syamsul Hadi

    2014-08-01

    Full Text Available Background Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are easily isolated, expanded and stored using standard techniques. Their potential for regenerative therapy of chronic wounds should be evaluated. The aim of this study was to determine allogeneic fibroblast viability in the presence of peripheral blood mononuclear cells (PBMC. Methods In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4 expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1 proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test. Results Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05. Calculated viable cell numbers increased 1.8 and 5.1-fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity Conclusions Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management.

  1. Allogeneic human dermal fibroblasts are viable in peripheral blood mononuclear co-culture

    Directory of Open Access Journals (Sweden)

    Restu Syamsul Hadi

    2015-12-01

    Full Text Available BACKGROUND Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are easily isolated, expanded and stored using standard techniques. Their potential for regenerative therapy of chronic wounds should be evaluated. The aim of this study was to determine allogeneic fibroblast viability in the presence of peripheral blood mononuclear cells (PBMC. METHODS In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4 expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1 proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test. RESULTS Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05. Calculated viable cell numbers increased 1.8 and 5.1- fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity CONCLUSIONS Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management.

  2. Small Sized Drone Fall Recover Mechanism Design

    Science.gov (United States)

    LIU, Tzu-Heng; CHAO, Fang-Lin; LIOU, Jhen-Yuan

    2017-12-01

    Drones uses four motors to rotate clockwise, counter-clockwise, or change in rotational speed to change its status of motion. The problem of Unmanned Aerial Vehicle turnover causes personal loses and harm local environment. Designs of devices that can let falling drones recover are discussed. The models attempt to change the orientation, so that the drone may be able to improve to the point where it can take off again. The design flow included looking for functional elements, using simplify model to estimate primary functional characteristics, and find the appropriate design parameters. For reducing the complexity, we adopted the simple rotate mechanism with rotating arms to change the fuselage angle and reduce the dependence on the extra-components. A rough model was built to verify structure, and then the concept drawing and prototype were constructed. We made the prototype through the integration of mechanical part and the electronic control circuit. The electronic control module that selected is Arduino-mini pro. Through the Bluetooth modules, user can start the rebound mechanism by the motor control signal. Protections frames are added around each propeller to improve the body rotate problem. Limited by current size of Arduino module, motor and rebound mechanism make the main chassis more massive than the commercial product. However, built-in sensor and circuit miniaturization will improve it in future.

  3. Social Networking and Smart Technology: Viable Environmental Communication Tools…?

    Science.gov (United States)

    Montain, J.; Byrne, J. M.

    2010-12-01

    To what extent do popular social networking channels represent a viable means for disseminating information regarding environmental change to the general public? Are new forms of communication such as YouTube™, Facebook™, MySpace™ and Twitter™ and smart devices such as iPhone™ and BlackBerry™ useful and effective in terms motivating people into social action and behavioural modification; or do they simply pay ‘lip service’ to these pressing environmental issues? This project will explore the background connections between social networking and environmental communication and education; and outline why such tools might be an appropriate way to connect to a broad audience in an efficient and unconventional manner. Further, research will survey the current prevalence of reliable environmental change information on social networking Internet-based media; and finally, suggestions for improved strategies and new directions will be provided.

  4. Dissolvable tattoo sensors: from science fiction to a viable technology

    Science.gov (United States)

    Cheng, Huanyu; Yi, Ning

    2017-01-01

    Early surrealistic painting and science fiction movies have envisioned dissolvable tattoo electronic devices. In this paper, we will review the recent advances that transform that vision into a viable technology, with extended capabilities even beyond the early vision. Specifically, we focus on the discussion of a stretchable design for tattoo sensors and degradable materials for dissolvable sensors, in the form of inorganic devices with a performance comparable to modern electronics. Integration of these two technologies as well as the future developments of bio-integrated devices is also discussed. Many of the appealing ideas behind developments of these devices are drawn from nature and especially biological systems. Thus, bio-inspiration is believed to continue playing a key role in future devices for bio-integration and beyond.

  5. A viable logarithmic f(R) model for inflation

    Energy Technology Data Exchange (ETDEWEB)

    Amin, M.; Khalil, S. [Center for Fundamental Physics, Zewail City of Science and Technology,6 October City, Giza (Egypt); Salah, M. [Center for Fundamental Physics, Zewail City of Science and Technology,6 October City, Giza (Egypt); Department of Mathematics, Faculty of Science, Cairo University,Giza (Egypt)

    2016-08-18

    Inflation in the framework of f(R) modified gravity is revisited. We study the conditions that f(R) should satisfy in order to lead to a viable inflationary model in the original form and in the Einstein frame. Based on these criteria we propose a new logarithmic model as a potential candidate for f(R) theories aiming to describe inflation consistent with observations from Planck satellite (2015). The model predicts scalar spectral index 0.9615

  6. Simultaneous pyometra and viable puppies’ gestation in a bitch

    Directory of Open Access Journals (Sweden)

    A. Risso

    2014-08-01

    Full Text Available Here we describe a case of pyometra coexisting with gestation in a 4.5 year-old miniature short-haired Dachshund. The dog exhibited depression, vaginal discharge, polydipsia and dehydration. Ultrasound examination revealed the presence of low to moderate anechoic fluid collection in the left uterine horn. Blood analysis revealed mild neutrophilia with a left shift. Based on these findings a presumptive diagnosis of pyometra was made and the bitch was treated using amoxicillin-clavulanate with dopaminergic agonist (cabergoline. A second ultrasound scan revealed the presence of two gestational vesicles in the right uterine horn that were successfully carried to term. Unusually, while pyometra persisted in the left uterine horn, two viable puppies were delivered by caesarean section from the right uterine horn.

  7. The viable but non-culturable state in pathogenic Escherichia coli: A general review

    Directory of Open Access Journals (Sweden)

    Jennifer A. Pienaar

    2016-05-01

    Objectives: This review discusses various general aspects of the VBNC state, the mechanisms and possible public health impact of indicator and pathogenic E. coli entering into the VBNC state. Method: A literature review was conducted to ascertain the possibleimpact of E. coli entering into the VBNC state. Results: Escherichia coli enter into the VBNC state by means of several induction mechanisms. Various authors have found that E. coli can be resuscitated post-VBNC. Certain strains of pathogenic E. coli are still able to produce toxins in the VBNC state, whilst others are avirulent during the VBNC state but are able to regain virulence after resuscitation. Conclusion: Pathogenic and indicator E. coli entering into the VBNC state could have an adverse effect on public health if conventional detection methods are used, where the number of viable cells could be underestimated and the VBNC cells still produce toxins or could, at anytime, be resuscitated and become virulent again.

  8. Profiling Total Viable Bacteria in a Hemodialysis Water Treatment System.

    Science.gov (United States)

    Chen, Lihua; Zhu, Xuan; Zhang, Menglu; Wang, Yuxin; Lv, Tianyu; Zhang, Shenghua; Yu, Xin

    2017-05-28

    Culture-dependent methods, such as heterotrophic plate counting (HPC), are usually applied to evaluate the bacteriological quality of hemodialysis water. However, these methods cannot detect the uncultured or viable but non-culturable (VBNC) bacteria, both of which may be quantitatively predominant throughout the hemodialysis water treatment system. Therefore, propidium monoazide (PMA)-qPCR associated with HPC was used together to profile the distribution of the total viable bacteria in such a system. Moreover, high-throughput sequencing of 16S rRNA gene amplicons was utilized to analyze the microbial community structure and diversity. The HPC results indicated that the total bacterial counts conformed to the standards, yet the bacteria amounts were abruptly enhanced after carbon filter treatment. Nevertheless, the bacterial counts detected by PMA-qPCR, with the highest levels of 2.14 × 10 7 copies/100 ml in softener water, were much higher than the corresponding HPC results, which demonstrated the occurrence of numerous uncultured or VBNC bacteria among the entire system before reverse osmosis (RO). In addition, the microbial community structure was very different and the diversity was enhanced after the carbon filter. Although the diversity was minimized after RO treatment, pathogens such as Escherichia could still be detected in the RO effluent. In general, both the amounts of bacteria and the complexity of microbial community in the hemodialysis water treatment system revealed by molecular approaches were much higher than by traditional method. These results suggested the higher health risk potential for hemodialysis patients from the up-to-standard water. The treatment process could also be optimized, based on the results of this study.

  9. Trait aggression and hostility in recovered alcoholics.

    Science.gov (United States)

    Ziherl, Slavko; Cebasek Travnik, Zdenka; Kores Plesnicar, Blanka; Tomori, Martina; Zalar, Bojan

    2007-01-01

    There is a long-recognized association between alcohol consumption and aggressive behavior. This study was designed to examine aggression in a group of socially well-adapted recovered alcoholics (RA). The question addressed was whether the treatment, together with long-term abstinence from alcohol, could reduce aggression and hostility in RA. A group of male RA (n = 64), who did not meet the DSM-IV criteria for any psychiatric or personality disorder, were recruited to the study from aftercare groups. According to data from their group therapists, they were reliably abstinent for at least 3 years and socially well adapted. The study participants representing the control group (n = 69), diagnosed as being 'reliable nonalcoholics' (NA) by the Munich Alcoholism Test, were recruited from general practice. Data were derived from an in-house questionnaire on general characteristics of both groups, and aggressive and hostility traits were assessed using the Buss-Durkee Hostility Inventory (BDHI). The univariate and multivariate between-groups design was used for data analysis. Taking into account the BDHI dimensions of aggression and hostility, the difference between RA and NA groups was statistically significant [Wilks' lambda (8, 125) = 0.769; p = 0.00004]. There were statistically significant differences in the BDHI scales for indirect aggression, irritability, negativism, suspicion, resentment, and guilt. Both RA and Na groups did not differ significantly in variables that assessed physical and verbal aggression. After a 3-year abstinence, subjects from the RA group displayed signs of hostility and covert aggression. Copyright (c) 2007 S. Karger AG, Basel.

  10. Methane Hydrates: More Than a Viable Aviation Fuel Feedstock Option

    Science.gov (United States)

    Hendricks, Robert C.

    2007-01-01

    Demand for hydrocarbon fuels is steadily increasing, and greenhouse gas emissions continue to rise unabated with the energy demand. Alternate fuels will be coming on line to meet that demand. This report examines the recovering of methane from methane hydrates for fuel to meet this demand rather than permitting its natural release into the environment, which will be detrimental to the planet. Some background on the nature, vast sizes, and stability of sedimentary and permafrost formations of hydrates are discussed. A few examples of the severe problems associated with methane recovery from these hydrates are presented along with the potential impact on the environment and coastal waters. Future availability of methane from hydrates may become an attractive option for aviation fueling, and so future aircraft design associated with methane fueling is considered.

  11. Workers’ Self-management, Recovered Companies and the Sociology of Work

    Science.gov (United States)

    Croucher, Richard

    2014-01-01

    We analyse how far Argentina’s worker-recovered companies (WRCs) have sustained themselves and their principles of equity and workers’ self-management since becoming widespread following the country’s 2001–2 economic crisis. Specialist Spanish-language sources, survey data and documents are analysed through four key sociological themes. We find that the number of WRCs has increased in Argentina, and that they represent a viable production model. Further, they have generally maintained their central principles and even flourished. This occurred despite the global economic crisis, legal and financial pressures to adopt capitalist practices and management structures, the risk of market absorption and state attempts to coopt, demobilise and depoliticise the movement. We argue that today they function as a much-needed international beacon of an alternative vision for labour and that integration of their experience has potential to revitalise the field. PMID:25419009

  12. Workers' Self-management, Recovered Companies and the Sociology of Work.

    Science.gov (United States)

    Ozarow, Daniel; Croucher, Richard

    2014-10-01

    We analyse how far Argentina's worker-recovered companies (WRCs) have sustained themselves and their principles of equity and workers' self-management since becoming widespread following the country's 2001-2 economic crisis. Specialist Spanish-language sources, survey data and documents are analysed through four key sociological themes. We find that the number of WRCs has increased in Argentina, and that they represent a viable production model. Further, they have generally maintained their central principles and even flourished. This occurred despite the global economic crisis, legal and financial pressures to adopt capitalist practices and management structures, the risk of market absorption and state attempts to coopt, demobilise and depoliticise the movement. We argue that today they function as a much-needed international beacon of an alternative vision for labour and that integration of their experience has potential to revitalise the field.

  13. [Bacterial diversity recovered from Qiyi Glacier and runoff, Qilian Mts].

    Science.gov (United States)

    Xie, Jun; Wang, Ning-Lian; Chen, Liang; Li, Quan-Lian; He, Jian-Qiao; Jiang, Xi; Wu, Xiao-Bo

    2009-09-15

    Abundance and species diversity of bacteria were investigated respectively by epifluorescence microscope, the culture method and the analysis of 16S rDNA genes, with snow and runoff samples from Qiyi Glacier in the Qilian Mts. According to the results, the total microbial cells and bacterial CFU range from 10(3) to 10(5) cells x mL(-1) and 0-600 cfu x mL(-1), respectively. The 16S rDNA gene of 22 bacterial isolates recovered from snow and runoff samples belong to following groups: Bacteroidetes, Actinobacteria, Firmicutes and alpha, beta, gamma-Proteobacteria. Based on their 16S rRNA sequences, Bacteroidetes forms the largest cluster in terms of abundance (80% of all isolates) and Pedobacter and Pseudomonas form the dominant genera in terms of abundance (90% of all isolates). Compared with bacteria revealed from ice and snow in Qinghai-Tibet Plateau, South Pole and North Pole which have been reported, the bacteria belonging to Pantoea, Providencia, Terrabacter, Aerococcus and Oxalobacteraceae are especially exist in Qiyi Glacier as far as we know.

  14. The elusive minimum viable population size for white sturgeon

    Energy Technology Data Exchange (ETDEWEB)

    Jager, Yetta [ORNL; Lepla, Ken B. [Idaho Power Company; Van Winkle, Webb [Van Windle Environmental Consulting; James, Mr Brad [Washington Department of Fish and Wildlife; McAdam, Dr Steve [University of British Columbia, Vancouver

    2010-01-01

    Biological conservation of sturgeon populations is a concern for many species. Those responsible for managing the white sturgeon (Acipenser transmontanus) and similar species are interested in identifying extinction thresholds to avoid. Two thresholds that exist in theory are the minimum viable population size (MVP) and minimum amount of suitable habitat. In this paper, we present both model and empirical estimates of these thresholds. We modified a population viability analysis (PVA) model for white sturgeon to include two new Allee mechanisms. Despite this, PVA-based MVP estimates were unrealistically low compared with empirical estimates unless opportunities for spawning were assumed to be less frequent. PVA results revealed a trade-off between MVP and habitat thresholds; smaller populations persisted in longer river segments and vice versa. Our empirical analyses suggested (1) a MVP range based on population trends from 1,194 to 27,700 individuals, and (2) a MVP estimate of 4,000 individuals based on recruitment. Long-term historical population surveys are needed for more populations to pinpoint an MVP based on trends, whereas the available data were sufficient to estimate MVP based on recruitment. Beyond the MVP, we developed a hierarchical model for population status based on empirical data. Metapopulation support was the most important predictor of population health, followed by the length of free-flowing habitat, with habitat thresholds at 26 and 150 km. Together, these results suggest that habitat and connectivity are important determinants of population status that likely influence the site-specific MVP thresholds.

  15. Towards a viable and just global nursing ethics.

    Science.gov (United States)

    Crigger, Nancy J

    2008-01-01

    Globalization, an outgrowth of technology, while informing us about people throughout the world, also raises our awareness of the extreme economic and social disparities that exist among nations. As part of a global discipline, nurses are vitally interested in reducing and eliminating disparities so that better health is achieved for all people. Recent literature in nursing encourages our discipline to engage more actively with social justice issues. Justice in health care is a major commitment of nursing; thus questions in the larger sphere of globalization, justice and ethics, are our discipline's questions also. Global justice, or fairness, is not an issue for some groups or institutions, but a deeper human rights issue that is a responsibility for everyone. What can we do to help reduce or eliminate the social and economic disparities that are so evident? What kind of ethical milieu is needed to address the threat that globalization imposes on justice and fairness? This article enriches the conceptualization of globalization by investigating recent work by Schweiker and Twiss. In addition, I discuss five qualities or characteristics that will facilitate the development of a viable and just global ethic. A global ethic guides all people in their response to human rights and poverty. Technology and business, two major forces in globalization that are generally considered beneficial, are critiqued as barriers to social justice and the common good.

  16. Keeping checkpoint/restart viable for exascale systems.

    Energy Technology Data Exchange (ETDEWEB)

    Riesen, Rolf E.; Bridges, Patrick G. (IBM Research, Ireland, Mulhuddart, Dublin); Stearley, Jon R.; Laros, James H., III; Oldfield, Ron A.; Arnold, Dorian (University of New Mexico, Albuquerque, NM); Pedretti, Kevin Thomas Tauke; Ferreira, Kurt Brian; Brightwell, Ronald Brian

    2011-09-01

    Next-generation exascale systems, those capable of performing a quintillion (10{sup 18}) operations per second, are expected to be delivered in the next 8-10 years. These systems, which will be 1,000 times faster than current systems, will be of unprecedented scale. As these systems continue to grow in size, faults will become increasingly common, even over the course of small calculations. Therefore, issues such as fault tolerance and reliability will limit application scalability. Current techniques to ensure progress across faults like checkpoint/restart, the dominant fault tolerance mechanism for the last 25 years, are increasingly problematic at the scales of future systems due to their excessive overheads. In this work, we evaluate a number of techniques to decrease the overhead of checkpoint/restart and keep this method viable for future exascale systems. More specifically, this work evaluates state-machine replication to dramatically increase the checkpoint interval (the time between successive checkpoint) and hash-based, probabilistic incremental checkpointing using graphics processing units to decrease the checkpoint commit time (the time to save one checkpoint). Using a combination of empirical analysis, modeling, and simulation, we study the costs and benefits of these approaches on a wide range of parameters. These results, which cover of number of high-performance computing capability workloads, different failure distributions, hardware mean time to failures, and I/O bandwidths, show the potential benefits of these techniques for meeting the reliability demands of future exascale platforms.

  17. Is Greenberg’s “Macro-Carib” viable?

    Directory of Open Access Journals (Sweden)

    Spike Gildea

    2007-08-01

    Full Text Available In his landmark work Language in the Americas, Greenberg (1987 proposed that Macro-Carib was one of the major low-level stocks of South America, which together with Macro-Panoan and Macro-Ge-Bororo were claimed to comprise the putative Ge-Pano-Carib Phylum. His Macro-Carib includes the isolates Andoke and Kukura, and the Witotoan, Peba-Yaguan, and Cariban families. Greenberg’s primary evidence came from person-marking paradigms in individual languages, plus scattered words from individual languages collected into 79 Macro-Carib ‘etymologies’ and another 64 Amerind ‘etymologies’. The goal of this paper is to re-evaluate Greenberg’s Macro-Carib claim in the light of the much more extensive and reliable language data that has become available largely since 1987. Based on full person-marking paradigms for Proto-Cariban, Yagua, Bora and Andoke, we conclude that Greenberg’s morphological claims are unfounded. For our lexical comparison, we created lexical lists for Proto-Cariban, Proto-Witotoan, Yagua and Andoke, for both Greenberg’s 143 putative etymologies and for the Swadesh 100 list. From both lists, a total of 23 potential cognates were found, but no consonantal correspondences were repeated even once. We conclude that our greatly expanded and improved database does not provide sufficient evidence to convince the skeptic that the Macro-Carib hypothesis is viable.

  18. Case-based anatomy teaching: a viable alternative?

    Science.gov (United States)

    Eseonu, Onyedikachi; Carachi, Robert; Brindley, Nicola

    2013-08-01

    Over the last two decades, there has been a decline in the amount of time available for anatomy teaching in the medical undergraduate curriculum, and new methods of anatomy teaching have been adopted for pragmatic reasons, with little evidence base to support their proposed educational benefits. This study seeks to establish the effect of a case-based teaching method on students' confidence in anatomy. Forty-three student volunteers in the clinical phase of the Glasgow medical course were given weekly anatomy teaching sessions based on clinical case presentations over 4 weeks. The students were given an anatomy test, and were asked to rate their confidence in their anatomy knowledge before and after the teaching sessions. There was a two-point increase in students' self-rated confidence, and a 10.9 per cent increase in average test score after the case-based anatomy teaching sessions. Both of these increases were statistically significant (p teaching was also highly rated by students, which may make it a viable option for the teaching of anatomy in the modern medical curriculum. © 2013 John Wiley & Sons Ltd.

  19. Protein design algorithms predict viable resistance to an experimental antifolate.

    Science.gov (United States)

    Reeve, Stephanie M; Gainza, Pablo; Frey, Kathleen M; Georgiev, Ivelin; Donald, Bruce R; Anderson, Amy C

    2015-01-20

    Methods to accurately predict potential drug target mutations in response to early-stage leads could drive the design of more resilient first generation drug candidates. In this study, a structure-based protein design algorithm (K* in the OSPREY suite) was used to prospectively identify single-nucleotide polymorphisms that confer resistance to an experimental inhibitor effective against dihydrofolate reductase (DHFR) from Staphylococcus aureus. Four of the top-ranked mutations in DHFR were found to be catalytically competent and resistant to the inhibitor. Selection of resistant bacteria in vitro reveals that two of the predicted mutations arise in the background of a compensatory mutation. Using enzyme kinetics, microbiology, and crystal structures of the complexes, we determined the fitness of the mutant enzymes and strains, the structural basis of resistance, and the compensatory relationship of the mutations. To our knowledge, this work illustrates the first application of protein design algorithms to prospectively predict viable resistance mutations that arise in bacteria under antibiotic pressure.

  20. Towards viable cosmological models of disformal theories of gravity

    Science.gov (United States)

    Sakstein, Jeremy

    2015-01-01

    The late-time cosmological dynamics of disformal gravity are investigated using dynamical systems methods. It is shown that in the general case there are no stable attractors that screen fifth forces locally and simultaneously describe a dark energy dominated universe. Viable scenarios have late-time properties that are independent of the disformal parameters and are identical to the equivalent conformal quintessence model. Our analysis reveals that configurations where the Jordan frame metric becomes singular are only reached in the infinite future, thus explaining the natural pathology resistance observed numerically by several previous works. The viability of models where this can happen is discussed in terms of both the cosmological dynamics and local phenomena. We identify a special parameter tuning such that there is a new fixed point that can match the presently observed dark energy density and equation of state. This model is unviable when the scalar couples to the visible sector but may provide a good candidate model for theories where only dark matter is disformally coupled.

  1. SMA actuators: a viable practical technology (Presentation Video)

    Science.gov (United States)

    Browne, Alan L.; Brown, Jeffrey; Hodgson, Darel E.

    2015-04-01

    Diverse products either based solely on or incorporating Shape Memory Alloys (SMA) have and are being made in a wide range of industries, and IP is being captured. Why then compared to SE (superelastic) Nitinol, and especially conventional technology, do so few ideas reach production? This presentation delves deeply into this topic in reaching the final assessment that SMA actuators are indeed now a viable practical technology. The presentation begins with an introduction to and description of the fundamental basis of SMA actuator technology. Examples of multiple commercially available geometric forms of SMA actuators are given and the functionalities that they provide are described. This is followed by examples of multiple commercial products incorporating such SMA actuators. Given that there are literally millions of commercial products incorporating conventional actuator technologies, indications are given as to why there are their less than 1000 that utilize SMA. Experience based challenges to the commercial use of SMA actuators are described. Besides having to compete with existing non-SMA technology which is quite mature additional challenges that are unique to SM actuators are indicated these including a wider than expected set of technical engineering problems and challenges and that a broader scope of dynamics is required.

  2. Comparative study of original recover and recover KL in separable non-negative matrix factorization for topic detection in Twitter

    Science.gov (United States)

    Prabandari, R. D.; Murfi, H.

    2017-07-01

    An increasing amount of information on social media such as Twitter requires an efficient way to find the topics so that the information can be well managed. One of an automated method for topic detection is separable non-negative matrix factorization (SNMF). SNMF assumes that each topic has at least one word that does not appear on other topics. This method uses the direct approach and gives polynomial-time complexity, while the previous methods are iterative approaches and have NP-hard complexity. There are three steps of SNMF algorithm, i.e. constructing word co-occurrences, finding anchor words, and recovering topics. In this paper, we examine two topic recover methods, namely original recover that is using algebraic manipulation and recover KL that using probability approach with Kullback-Leibler divergence. Our simulations show that recover KL provides better accuracies in term of topic recall than original recover.

  3. Lactobacillus backii and Pediococcus damnosus isolated from 170-year-old beer recovered from a shipwreck lack the metabolic activities required to grow in modern lager beer.

    Science.gov (United States)

    Kajala, Ilkka; Bergsveinson, Jordyn; Friesen, Vanessa; Redekop, Anna; Juvonen, Riikka; Storgårds, Erna; Ziola, Barry

    2017-11-08

    In 2010, bottles of beer containing viable bacteria of the common beer-spoilage species Lactobacillus backii and Pediococcus damnosus were recovered from a shipwreck near the Åland Islands, Finland. The 170-year quiescent state maintained by the shipwreck bacteria presented a unique opportunity to study lactic acid bacteria (LAB) evolution vis a vis growth and survival in the beer environment. Three shipwreck bacteria (one L. backii strain and two P. damnosus strains) and modern-day beer-spoilage isolates of the same two species were genome sequenced, characterized for hop iso-α-acid tolerance, and growth in degassed lager and wheat beer. In addition, plasmid variants of the modern-day P. damnosus strain were analyzed for the effect of plasmid-encoded genes on growth in lager beer. Coding content on two plasmids was identified as essential for LAB growth in modern lager beer. Three chromosomal regions containing genes related to sugar transport and cell wall polysaccharides were shared by pediococci able to grow in beer. Our results show that the three shipwreck bacteria lack the necessary plasmid-located genetic content to grow in modern lager beer, but carry additional genes related to acid tolerance and biofilm formation compared to their modern counterparts. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  4. The viable but non-culturable state in pathogenic Escherichia coli: A general review

    Directory of Open Access Journals (Sweden)

    Jennifer A. Pienaar

    2016-02-01

    Full Text Available Background: The persistence and pathogenicity of pathogenic bacteria are dependent on the ability of the species to survive in adverse conditions. During the infectious process, the organism may need to pass through certain hostile anatomical sites, such as the stomach. Under various environmental stresses, many bacteria enter into the viable but non-culturable (VBNC state, where they are ‘alive’ or metabolically active, but will not grow on conventional media. Escherichia coli bacteria encounter several diverse stress factors during their growth, survival and infection and thus may enter into the VBNC state.Objectives: This review discusses various general aspects of the VBNC state, the mechanisms and possible public health impact of indicator and pathogenic E. coli entering into the VBNC state.Method: A literature review was conducted to ascertain the possibleimpact of E. coli entering into the VBNC state.Results: Escherichia coli enter into the VBNC state by means of several induction mechanisms. Various authors have found that E. coli can be resuscitated post-VBNC. Certain strains of pathogenic E. coli are still able to produce toxins in the VBNC state, whilst others are avirulent during the VBNC state but are able to regain virulence after resuscitation.Conclusion: Pathogenic and indicator E. coli entering into the VBNC state could have an adverse effect on public health if conventional detection methods are used, where the number of viable cells could be underestimated and the VBNC cells still produce toxins or could, at anytime, be resuscitated and become virulent again.

  5. Acupuntura un tratamiento viable para las adicciones en Colombia

    Directory of Open Access Journals (Sweden)

    Hernán López-Suescún

    2013-09-01

    Full Text Available Resumen La acupuntura es una antigua técnica terapéutica desarrollada en China, que ha evidenciado ser efectiva en síntomas como las náuseas, vómito y dolor dentario. A pesar del sustento fisiológico que posibilitaría un uso efectivo en otras patologías, incluyendo el campo de las adicciones, los estudios son contradictorios, posiblemente por la diferencias de visión entre la medicina oriental y la occidental. El consumo de psicoactivos es un problema de salud pública en Colombia y en el mundo que genera grandes costos tangibles e intangibles, los cuales, en países desarrollados, puede llegar hasta el 1,6 % del PIB. En contraste, el beneficio económico del tratamiento de las adicciones, según las Naciones Unidas Contra la Droga y el Delito (UNODC, está entre 1:3 a 1:13; por lo tanto, cualquier esfuerzo que se realice en favor de los consumidores es una ganancia. Con base en estos datos, los organismos internacionales han generado políticas que ayudan a aminorar estos efectos. Colombia, como integrante de estos organismos, ha realizado varios compromisos para llevar a cabo dichas metas. Los tratamientos con auriculoterapia, como el protocolo NADA (National Acupuncture Detoxification Association, son los métodos más usados para las adicciones en el mundo, y aunque no se ha logrado evidenciar su efectividad, por su costo, facilidad y el poco riesgo de efectos adversos se hace viable en un país con pocos recursos económicos como Colombia.

  6. Projecting recovered paper supply in the United States.

    Science.gov (United States)

    James L. Howard; Ken Skog; Peter J. Ince

    2002-01-01

    The use of recovered paper has an influence on the amount of timber harvested as well as the feasibility of expanded recycling programs. During the 1990’s markets and prices for recovered materials have fluctuated widely. There are a number of factors driving these market changes including economic conditions domestically and internationally. The objective of this...

  7. Evaluation of an automated struvite reactor to recover phosphorus ...

    African Journals Online (AJOL)

    In the present study we attempted to develop a reactor system to recover phosphorus by struvite precipitation, and which can be installed anywhere in the field without access to a laboratory. A reactor was developed that can run fully automated and recover up to 93% of total phosphorus (total P). Turbidity and conductivity ...

  8. 27 CFR 20.213 - Reuse of recovered spirits.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Reuse of recovered spirits. 20.213 Section 20.213 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU... Alcohol, Specially Denatured Rum, or Articles § 20.213 Reuse of recovered spirits. (a) If the denatured...

  9. 48 CFR 245.607-2 - Recovering precious metals.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 3 2010-10-01 2010-10-01 false Recovering precious metals... Disposal of Contractor Inventory 245.607-2 Recovering precious metals. (b) Precious metals are silver, gold... office with disposition instructions for certain categories of precious metals-bearing property...

  10. 48 CFR 945.607-2 - Recovering precious metals.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false Recovering precious metals... Recovering precious metals. (b) Contractors generating contractor inventory containing precious metals shall... the DOE precious metals pool. This includes all precious metals in any form, including shapes, scrap...

  11. A propidium monoazide–quantitative PCR method for the detection and quantification of viable Enterococcus faecalis in large-volume samples of marine waters

    KAUST Repository

    Salam, Khaled W.

    2014-08-23

    The development of rapid detection assays of cell viability is essential for monitoring the microbiological quality of water systems. Coupling propidium monoazide with quantitative PCR (PMA-qPCR) has been successfully applied in different studies for the detection and quantification of viable cells in small-volume samples (0.25-1.00 mL), but it has not been evaluated sufficiently in marine environments or in large-volume samples. In this study, we successfully integrated blue light-emitting diodes for photoactivating PMA and membrane filtration into the PMA-qPCR assay for the rapid detection and quantification of viable Enterococcus faecalis cells in 10-mL samples of marine waters. The assay was optimized in phosphate-buffered saline and seawater, reducing the qPCR signal of heat-killed E. faecalis cells by 4 log10 and 3 log10 units, respectively. Results suggest that high total dissolved solid concentration (32 g/L) in seawater can reduce PMA activity. Optimal PMA-qPCR standard curves with a 6-log dynamic range and detection limit of 102 cells/mL were generated for quantifying viable E. faecalis cells in marine waters. The developed assay was compared with the standard membrane filter (MF) method by quantifying viable E. faecalis cells in seawater samples exposed to solar radiation. The results of the developed PMA-qPCR assay did not match that of the standard MF method. This difference in the results reflects the different physiological states of E. faecalis cells in seawater. In conclusion, the developed assay is a rapid (∼5 h) method for the quantification of viable E. faecalis cells in marine recreational waters, which should be further improved and tested in different seawater settings. © 2014 Springer-Verlag Berlin Heidelberg.

  12. Detection of Only Viable Bacterial Spores Using a Live/Dead Indicator in Mixed Populations

    Science.gov (United States)

    Behar, Alberto E.; Stam, Christina N.; Smiley, Ronald

    2013-01-01

    This method uses a photoaffinity label that recognizes DNA and can be used to distinguish populations of bacterial cells from bacterial spores without the use of heat shocking during conventional culture, and live from dead bacterial spores using molecular-based methods. Biological validation of commercial sterility using traditional and alternative technologies remains challenging. Recovery of viable spores is cumbersome, as the process requires substantial incubation time, and the extended time to results limits the ability to quickly evaluate the efficacy of existing technologies. Nucleic acid amplification approaches such as PCR (polymerase chain reaction) have shown promise for improving time to detection for a wide range of applications. Recent real-time PCR methods are particularly promising, as these methods can be made at least semi-quantitative by correspondence to a standard curve. Nonetheless, PCR-based methods are rarely used for process validation, largely because the DNA from dead bacterial cells is highly stable and hence, DNA-based amplification methods fail to discriminate between live and inactivated microorganisms. Currently, no published method has been shown to effectively distinguish between live and dead bacterial spores. This technology uses a DNA binding photoaffinity label that can be used to distinguish between live and dead bacterial spores with detection limits ranging from 109 to 102 spores/mL. An environmental sample suspected of containing a mixture of live and dead vegetative cells and bacterial endospores is treated with a photoaffinity label. This step will eliminate any vegetative cells (live or dead) and dead endospores present in the sample. To further determine the bacterial spore viability, DNA is extracted from the spores and total population is quantified by real-time PCR. The current NASA standard assay takes 72 hours for results. Part of this procedure requires a heat shock step at 80 degC for 15 minutes before the

  13. Coscheduling in Clusters: Is It a Viable Alternative?

    Energy Technology Data Exchange (ETDEWEB)

    Choi, G S; Kim, J H; Ersoz, D; Yoo, A B; Das, C R

    2003-11-10

    than spin-based techniques like PB on a Linux platform. Third, the proposed HYBRID scheduling provides the best performance-energy behavior and can be implemented on any cluster with little effort. All these results suggest that blocking-based coscheduling techniques are viable candidates to be used instead of batching scheme for significant performance-energy benefits.

  14. Expression of adhesion molecules, chemokines and matrix metallo- proteinases (MMPs) in viable and degenerating stage of Taenia solium metacestode in swine neurocysticercosis.

    Science.gov (United States)

    Singh, Satyendra K; Singh, Aloukick K; Prasad, Kashi N; Singh, Amrita; Singh, Avinash; Rai, Ravi P; Tripathi, Mukesh; Gupta, Rakesh K; Husain, Nuzhat

    2015-11-30

    Neurocysticercosis (NCC) is a parasitic infection of central nervous system (CNS). Expression of adhesion molecules, chemokines and matrix metalloproteinases (MMPs) were investigated on brain tissues surrounding viable (n=15) and degenerating cysticerci (n=15) of Taenia solium in swine by real-time RT-PCR and ELISA. Gelatin gel zymography was performed for MMPs activity. ICAM-1 (intercellular adhesion molecule-1), E-selectin, MIP-1α (macrophage inflammatory protein-1α), Eotaxin-1 and RANTES (regulated on activation, normal T cell expressed and secreted) were associated with degenerating cysticerci (cysts). However, VCAM-1 (vascular cell adhesion molecule-1), MCP-1 (monocyte chemotactic protein-1), MMP-2 and MMP-9 were associated with both viable and degenerating cysts. In conclusion, viable and degenerating cysticerci have different immune molecule profiles and role of these molecules in disease pathogenesis needs to be investigated. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Detection and quantification of viable Bacillus cereus group species in milk by propidium monoazide quantitative real-time PCR.

    Science.gov (United States)

    Cattani, Fernanda; Barth, Valdir C; Nasário, Jéssica S R; Ferreira, Carlos A S; Oliveira, Sílvia D

    2016-04-01

    The Bacillus cereus group includes important spore-forming bacteria that present spoilage capability and may cause foodborne diseases. These microorganisms are traditionally evaluated in food using culturing methods, which can be laborious and time-consuming, and may also fail to detect bacteria in a viable but nonculturable state. The purpose of this study was to develop a quantitative real-time PCR (qPCR) combined with a propidium monoazide (PMA) treatment to analyze the contamination of UHT milk by B. cereus group species viable cells. Thirty micrograms per milliliter of PMA was shown to be the most effective concentration for reducing the PCR amplification of extracellular DNA and DNA from dead cells. The quantification limit of the PMA-qPCR assay was 7.5 × 10(2) cfu/mL of milk. One hundred thirty-five UHT milk samples were analyzed to evaluate the association of PMA to qPCR to selectively detect viable cells. The PMA-qPCR was able to detect B. cereus group species in 44 samples (32.6%), whereas qPCR without PMA detected 78 positive samples (57.8%). Therefore, the PMA probably inhibited the amplification of DNA from cells that were killed during UHT processing, which avoided an overestimation of bacterial cells when using qPCR and, thus, did not overvalue potential health risks. A culture-based method was also used to detect and quantify B. cereus sensu stricto in the same samples and showed positive results in 15 (11.1%) samples. The culture method and PMA-qPCR allowed the detection of B. cereus sensu stricto in quantities compatible with the infective dose required to cause foodborne disease in 3 samples, indicating that, depending on the storage conditions, even after UHT treatment, infective doses may be reached in ready-to-consume products. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  16. Eradication of high viable loads of Listeria monocytogenes contaminating food-contact surfaces

    Directory of Open Access Journals (Sweden)

    Silvia ede Candia

    2015-07-01

    Full Text Available This study demonstrates the efficacy of cold gaseous ozone treatments at low concentrations in the eradication of high Listeria monocytogenes viable cell loads from glass, polypropylene, stainless steel and expanded polystyrene food-contact surfaces. Using a step by step approach, involving the selection of the most resistant strain-surface combinations, 11 Listeria spp. strains resulted inactivated by a continuous ozone flow at 1.07 mg m-3 after 24 or 48 h of cold incubation, depending on both strain and surface evaluated. Increasing the inoculum level to 9 log CFU coupon-1, the best inactivation rate was obtained after 48h of treatment at 3.21 mg m-3 ozone concentration when cells were deposited onto stainless steel and expanded polystyrene coupons, resulted the most resistant food-contact surfaces in the previous assays.The addition of naturally microbiologically contaminated meat extract to a high load of L. monocytogenes LMG 23775 cells, the most resistant strain out of the 11 assayed Listeria spp. strains, led to its complete inactivation after four days of treatment.To the best of our knowledge, this is the first report describing the survival of L. monocytogenes and the effect of ozone treatment under cold storage conditions on expanded polystyrene, a commonly-used material in food packaging. These results could be useful for reducing pathogen cross-contamination phenomena during cold food storage.

  17. Colpoda secrete viable Listeria monocytogenes within faecal pellets.

    Science.gov (United States)

    Raghu Nadhanan, Rethish; Thomas, Connor J

    2014-02-01

    Transmission electron microscopy was used to demonstrate that co-cultures of the ciliate Colpoda RR (an environmental isolate) and Colpoda MLS-5 (a food processing environment isolate) with the pathogenic Listeria monocytogenes DRDC8 resulted in secretion of faecal pellets containing intact DRDC8 cells. A green fluorescent protein expressing variant of DRDC8 was used in co-cultures to confirm that the pellet-associated bacterial cells were L. monocytogenes. Viability was confirmed by plate counts, and assay of microbial respiratory activity-proved DRDC8 cells present within faecal pellets was metabolically active. Following treatment of faecal pellets secreted by Colpoda RR and MLS-5 with gentamycin and sodium hypochlorite (NaOCl), no loss of viability of the pellet-located DRDC8 cells was observed, indicating that faecal pellet encapsulated DRDC8 cells are resistant to biocidal agents. This work suggests that Colpoda-derived faecal pellets may provide a mechanism for transmission of L. monocytogenes and other pathogenic bacteria. Furthermore, bacteria encapsulated by faecal pellets may be resistant to disinfectants and cleaning agents used in food manufacturing and preparation facilities. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

  18. Effect of myocardial revascularisation on left ventricular systolic function in patients with and without viable myocardium: should non-viable segments be revascularised?

    Science.gov (United States)

    Stipac, Alja Vlahovic; Stankovic, Ivan; Vidakovic, Radosav; Putnikovic, Biljana; Ilic, Ivan; Milicic, Biljana; Neskovic, Aleksandar N

    2013-12-01

    To assess the effect of surgical revascularisation on left ventricular (LV) systolic function in patients with viable and non-viable dysfunctional LV segments determined by low dose dobutamine stress echocardiography (DSE). Prospective observational cohort study. Single tertiary care centre. Consecutive patients referred to surgical revascularisation (n=115). DSE and surgical revascularisation. Functional recovery defined as increase in ejection fraction ≥ 5% 1 year after revascularisation in patients with and without viable myocardium (viability defined as improvement of contractility in ≥ 4 LV segments on DSE). The mean age, ejection fraction and wall motion score index (WMSi) of patients were 59 ± 9 years, 44 ± 9% and 1.82 ± 0.31, respectively. There was no difference between DSE positive and DSE negative patients for any of those parameters at baseline study (p>0.05 for all). After 12 months, the ejection fraction increased 11 ± 1% in patients with viable myocardium vs 7 ± 1% in patients without viable myocardium (p=0.002). Moreover, in patients with viable myocardium, the greatest increase of ejection fraction occurred 1 month after surgery (9 ± 1%), whereas in those patients with negative DSE the ejection fraction increased more gradually (2±1% after 1 month, p=0.002 between groups for 1 month vs preoperative value), but still improved after 12 months follow-up (pmyocardial revascularisation. Functional recovery continuously occurs throughout the first year after surgical treatment.

  19. A technique for determining viable military logistics support alternatives

    Science.gov (United States)

    Hester, Jesse Stuart

    A look at today's US military will see them operating much beyond the scope of protecting and defending the United States. These operations now consist of, but are not limited to humanitarian aid, disaster relief, peace keeping, and conflict resolution. This broad spectrum of operational environments has necessitated a transformation of the individual military services to a hybrid force that is attempting to leverage the inherent and emerging capabilities and strengths of all those under the umbrella of the Department of Defense (DOD), this concept has been coined Joint Operations. Supporting Joint Operations requires a new approach to determining a viable military logistics support system. The logistics architecture for these operations has to accommodate scale, time, varied mission objectives, and imperfect information. Compounding the problem is the human in the loop (HITL) decision maker (DM) who is a necessary component for quickly assessing and planning logistics support activities. Past outcomes are not necessarily good indicators of future results, but they can provide a reasonable starting point for planning and prediction of specific needs for future requirements. Adequately forecasting the necessary logistical support structure and commodities needed for any resource intensive environment has progressed well beyond stable demand assumptions to one in which dynamic and nonlinear environments can be captured with some degree of fidelity and accuracy. While these advances are important, a holistic approach that allows exploration of the operational environment or design space does not exist to guide the military logistician in a methodical way to support military forecasting activities. To bridge this capability gap, a method called Adaptive Technique for Logistics Architecture Solutions (ATLAS) has been developed. This method provides a process that facilitates the use of techniques and tools that filter and provide relevant information to the DM. By doing

  20. Lipogenic metabolism: a viable target for prostate cancer treatment?

    Directory of Open Access Journals (Sweden)

    Mengmeng Liang

    2014-10-01

    Full Text Available Cancer cells often depend on altered metabolism compared with their normal counterparts. [1],[2],[3],[4] As observed in 1924 by Otto Warburg, cancer cells show preferential glucose consumption by way of aerobic glycolysis while normal cells generally assume mitochondrial oxidative phosphorylation. [4] Another metabolic hallmark of carcinogenesis is altered lipid metabolism, whereby cancer cells may adopt enhanced de novo lipid production (lipogenesis. [1],[2],[3] Enhanced lipid metabolism is also observed in individuals with metabolic syndromes potentially a consequence of increasing popularity of the Standard American Diet, composed of high levels of saturated fats and carbohydrates. [5] A growing body of epidemiological data indicates a positive correlation between the occurrence of metabolic syndromes, such as cardiovascular disease, obesity, type-2 diabetes and associated hyperinsulemia, with the aggressiveness of cancer. [6],[7],[8],[9] Remarkably, it is estimated that for every 1% reduction in saturated fats, replaced by polyunsaturated, there would be a 2%-3% reduction in cardiovascular disease. [10] Thus, it is conceivable that an equally remarkable attenuation in cancer progression might be achieved with such a reduction in lipid accumulation.

  1. Improving CO2 Efficiency for Recovering Oil in Heterogeneous Reservoirs

    Energy Technology Data Exchange (ETDEWEB)

    Grigg, Reid B.; Svec, Robert K.

    2003-03-10

    The work strived to improve industry understanding of CO2 flooding mechanisms with the ultimate goal of economically recovering more of the U.S. oil reserves. The principle interests are in the related fields of mobility control and injectivity.

  2. Regeneration of viable oil palm plants from protoplasts by optimizing media components, growth regulators and cultivation procedures.

    Science.gov (United States)

    Masani, Mat Yunus Abdul; Noll, Gundula; Parveez, Ghulam Kadir Ahmad; Sambanthamurthi, Ravigadevi; Prüfer, Dirk

    2013-09-01

    Oil palm protoplasts are suitable as a starting material for the production of oil palm plants with new traits using approaches such as somatic hybridization, but attempts to regenerate viable plants from protoplasts have failed thus far. Here we demonstrate, for the first time, the regeneration of viable plants from protoplasts isolated from cell suspension cultures. We achieved a protoplast yield of 1.14×10(6) per gram fresh weight with a viability of 82% by incubating the callus in a digestion solution comprising 2% cellulase, 1% pectinase, 0.5% cellulase onuzuka R10, 0.1% pectolyase Y23, 3% KCl, 0.5% CaCl2 and 3.6% mannitol. The regeneration of protoplasts into viable plants required media optimization, the inclusion of plant growth regulators and the correct culture technique. Microcalli derived from protoplasts were obtained by establishing agarose bead cultures using Y3A medium supplemented with 10μM naphthalene acetic acid, 2μM 2,4-dichlorophenoxyacetic acid, 2μM indole-3-butyric acid, 2μM gibberellic acid and 2μM 2-γ-dimethylallylaminopurine. Small plantlets were regenerated from microcalli by somatic embryogenesis after successive subculturing steps in medium with limiting amounts of growth regulators supplemented with 200mg/l ascorbic acid. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  3. Articular Cartilage Repair Using Marrow Stimulation Augmented with a Viable Chondral Allograft: 9-Month Postoperative Histological Evaluation

    Directory of Open Access Journals (Sweden)

    James K. Hoffman

    2015-01-01

    Full Text Available Marrow stimulation is frequently employed to treat focal chondral defects of the knee. However, marrow stimulation typically results in fibrocartilage repair tissue rather than healthy hyaline cartilage, which, over time, predisposes the repair to failure. Recently, a cryopreserved viable chondral allograft was developed to augment marrow stimulation. The chondral allograft is comprised of native viable chondrocytes, chondrogenic growth factors, and extracellular matrix proteins within the superficial, transitional, and radial zones of hyaline cartilage. Therefore, host mesenchymal stem cells that infiltrate the graft from the underlying bone marrow following marrow stimulation are provided with the optimal microenvironment to undergo chondrogenesis. The present report describes treatment of a trochlear defect with marrow stimulation augmented with this novel chondral allograft, along with nine month postoperative histological results. At nine months, the patient demonstrated complete resolution of pain and improvement in function, and the repair tissue consisted of 85% hyaline cartilage. For comparison, a biopsy obtained from a patient 8.2 months after treatment with marrow stimulation alone contained only 5% hyaline cartilage. These outcomes suggest that augmenting marrow stimulation with the viable chondral allograft can eliminate pain and improve outcomes, compared with marrow stimulation alone.

  4. Recovering heavy rare earth metals from magnet scrap

    Science.gov (United States)

    Ott, Ryan T.; McCallum, Ralph W.; Jones, Lawrence L.

    2017-08-08

    A method of treating rare earth metal-bearing permanent magnet scrap, waste or other material in a manner to recover the heavy rare earth metal content separately from the light rare earth metal content. The heavy rare earth metal content can be recovered either as a heavy rare earth metal-enriched iron based alloy or as a heavy rare earth metal based alloy.

  5. Recovering a Basic Space from Issue Scales in R

    Directory of Open Access Journals (Sweden)

    Keith T. Poole

    2016-03-01

    Full Text Available basicspace is an R package that conducts Aldrich-McKelvey and Blackbox scaling to recover estimates of the underlying latent dimensions of issue scale data. We illustrate several applications of the package to survey data commonly used in the social sciences. Monte Carlo tests demonstrate that the procedure can recover latent dimensions and reproduce the matrix of responses at moderate levels of error and missing data.

  6. Real-time detection of viable microorganisms by intracellular phototautomerism

    Directory of Open Access Journals (Sweden)

    Schuren Frank

    2010-06-01

    Full Text Available Abstract Background To date, the detection of live microorganisms present in the environment or involved in infections is carried out by enumeration of colony forming units on agar plates, which is time consuming, laborious and limited to readily cultivable microorganisms. Although cultivation-independent methods are available, they involve multiple incubation steps and do mostly not discriminate between dead or live microorganisms. We present a novel generic method that is able to specifically monitor living microorganisms in a real-time manner. Results The developed method includes exposure of cells to a weak acid probe at low pH. The neutral probe rapidly permeates the membrane and enters the cytosol. In dead cells no signal is obtained, as the cytosolic pH reflects that of the acidic extracellular environment. In live cells with a neutral internal pH, the probe dissociates into a fluorescent phototautomeric anion. After reaching peak fluorescence, the population of live cells decays. This decay can be followed real-time as cell death coincides with intracellular acidification and return of the probe to its uncharged non-fluorescent state. The rise and decay of the fluorescence signal depends on the probe structure and appears discriminative for bacteria, fungi, and spores. We identified 13 unique probes, which can be applied in the real-time viability method described here. Under the experimental conditions used in a microplate reader, the reported method shows a detection limit of 106 bacteria ml-1, while the frequently used LIVE/DEAD BacLight™ Syto9 and propidium iodide stains show detection down to 106 and 107 bacteria ml-1, respectively. Conclusions We present a novel fluorescence-based method for viability assessment, which is applicable to all bacteria and eukaryotic cell types tested so far. The RTV method will have a significant impact in many areas of applied microbiology including research on biocidal activity, improvement of

  7. Process for recovering alkali metals and sulfur from alkali metal sulfides and polysulfides

    Science.gov (United States)

    Gordon, John Howard; Alvare, Javier

    2016-10-25

    Alkali metals and sulfur may be recovered from alkali monosulfide and polysulfides in an electrolytic process that utilizes an electrolytic cell having an alkali ion conductive membrane. An anolyte solution includes an alkali monosulfide, an alkali polysulfide, or a mixture thereof and a solvent that dissolves elemental sulfur. A catholyte includes molten alkali metal. Applying an electric current oxidizes sulfide and polysulfide in the anolyte compartment, causes alkali metal ions to pass through the alkali ion conductive membrane to the catholyte compartment, and reduces the alkali metal ions in the catholyte compartment. Liquid sulfur separates from the anolyte solution and may be recovered. The electrolytic cell is operated at a temperature where the formed alkali metal and sulfur are molten.

  8. Birth of viable puppies derived from breeding cloned female dogs with a cloned male.

    Science.gov (United States)

    Park, J E; Hong, S G; Kang, J T; Oh, H J; Kim, M K; Kim, M J; Kim, H J; Kim, D Y; Jang, G; Lee, B C

    2009-09-15

    Since the establishment of production of viable cloned dogs by somatic cell nucleus transfer, great concern has been given to the reproductive abilities of these animals (Canis familiaris). Therefore, we investigated reproductive activity of cloned dogs by (1) performing sperm analysis using computer-assisted sperm analysis and early embryonic development, (2) assessing reproductive cycling by measuring serum progesterone (P4) levels and performing vaginal cytology, and (3) breeding cloned dogs using artificial insemination. Results showed that most parameters of sperm motility in a cloned male dog were within the reference range, and in vivo-matured oocytes from a noncloned female were successfully fertilized by spermatozoa from a cloned male dog and develop normally to the 8-cell stage. Three cloned female dogs displayed normal patterns of P4 levels and morphologic changes of the vaginal epithelium. Two cloned female dogs became pregnant using semen from a cloned male dog and successfully delivered 10 puppies by natural labor. In conclusion, these data demonstrated that both cloned male and female dogs are fertile, and their puppies are currently alive and healthy with normal growth patterns.

  9. The Role of Mitochondria from Mature Oocyte to Viable Blastocyst

    Directory of Open Access Journals (Sweden)

    Scott Chappel

    2013-01-01

    Full Text Available The oocyte requires a vast supply of energy after fertilization to support critical events such as spindle formation, chromatid separation, and cell division. Until blastocyst implantation, the developing zygote is dependent on the existing pool of mitochondria. That pool size within each cell decreases with each cell division. Mitochondria obtained from oocytes of women of advanced reproductive age harbor DNA deletions and nucleotide variations that impair function. The combination of lower number and increased frequency of mutations and deletions may result in inadequate mitochondrial activity necessary for continued embryo development and cause pregnancy failure. Previous reports suggested that mitochondrial activity within oocytes may be supplemented by donor cytoplasmic transfer at the time of intracytoplasmic sperm injection (ICSI. Those reports showed success; however, safety concerns arose due to the potential of two distinct populations of mitochondrial genomes in the offspring. Mitochondrial augmentation of oocytes is now reconsidered in light of our current understanding of mitochondrial function and the publication of a number of animal studies. With a better understanding of the role of this organelle in oocytes immediately after fertilization, blastocyst and offspring, mitochondrial augmentation may be reconsidered as a method to improve oocyte quality.

  10. Commercializing Biorefinery Technology: A Case for the Multi-Product Pathway to a Viable Biorefinery

    Directory of Open Access Journals (Sweden)

    Shijie Liu

    2011-11-01

    Full Text Available While there may be many reasons why very interesting science ideas never reach commercial practice, one of the more prevalent is that the reaction or process, which is scientifically possible, cannot be made efficient enough to achieve economic viability. One pathway to economic viability for many business sectors is the multi-product portfolio. Research, development, and deployment of viable biorefinery technology must meld sound science with engineering and business economics. It is virtually axiomatic that increased value can be generated by isolating relatively pure substances from heterogeneous raw materials. Woody biomass is a heterogeneous raw material consisting of the major structural components, cellulose, lignin, and hemicelluloses, as well as minor components, such as extractives and ash. Cellulose is a linear homopolymer of D-glucopyrano-units with β-D(1®4 connections and is the wood component most resistant to chemical and biological degradation. Lignin is a macromolecule of phenylpropanoid units, second to cellulose in bio-resistance, and is the key component that is sought for removal from woody biomass in chemical pulping. Hemicelluloses are a collection of heteropolysaccharides, comprised mainly of 5- and 6-carbon sugars. Extractives, some of which have high commercial value, are a collection of low molecular weight organic and inorganic woody materials that can be removed, to some extent, under mild conditions. Applied Biorefinery Sciences, LLC (a private, New York, USA based company is commercializing a value-optimization pathway (the ABS Process™ for generating a multi-product portfolio by isolating and recovering homogeneous substances from each of the above mentioned major and minor woody biomass components. The ABS Process™ incorporates the patent pending, core biorefinery technology, “hot water extraction”, as developed at the State University of New York College of Environmental Science and Forestry (SUNY

  11. Cybernetically sound organizational structures II: Relating de Sitter's design theory to Beer's viable system model

    NARCIS (Netherlands)

    Achterbergh, J.M.I.M.; Vriens, D.J.

    2011-01-01

    - Purpose – The purpose of this paper is to show how the viable system model (VSM) and de Sitter's design theory can complement each other in the context of the diagnosis and design of viable organizations. - Design/methodology/approach – Key concepts from Beer's model and de Sitter's design

  12. Hymenolepis nana: immunity against oncosphere challenge in mice previously given viable or non-viable oncospheres of H. nana, H. diminuta, H. microstoma and Taenia taeniaeformis.

    Science.gov (United States)

    Ito, A; Onitake, K; Sasaki, J; Takami, T

    1991-04-01

    When mice, previously given oral inoculation with viable oncospheres of the heterologous cestode species (Hymenolepis diminuta, H. microstoma, Taenia taeniaeformis) and the homologous one (H. nana), were challenged with oncospheres of H. nana 4 days after the primary inoculation, they showed strong and complete resistance to H. nana challenge, respectively. However, the resistance was not evoked in mice given either infective eggs of Toxocara canis or non-viable oncospheres of all cestode species examined. Congenitally athymic nude mice given viable oncospheres did not show any resistance to H. nana either. Eosinophil infiltration around cysticercoids of H. nana in the intestinal villi appeared to be more prominent in mice previously given viable oncospheres of H. diminuta than in mice given non-viable oncospheres or PBS only. Some of the eosinophils in the villus harboring cysticercoid(s) of H. nana invaded the epithelia in the former, whereas all eosinophils remained in the lamina propria in the latter. There was almost no eosinophil infiltration in nude mice. Microscopic observations revealed that oncospheres of H. diminuta, which require beetles as the intermediate host like H. microstoma, could invade the mouse intestinal tissue. Therefore, it is strongly suggested that the strong cross resistance to H. nana in mice, induced by oncospheres of all heterologous cestode species, is thymus-dependent and due to oncospheral invasion into the intestinal tissue of mice.

  13. A Rapid Method for Quantifying Viable Mycobacterium avium subsp. paratuberculosis in Cellular Infection Assays

    Science.gov (United States)

    Pooley, Hannah B.; de Silva, Kumudika; Purdie, Auriol C.; Begg, Douglas J.; Whittington, Richard J.

    2016-01-01

    ABSTRACT Determining the viability of bacteria is a key outcome of in vitro cellular infection assays. Currently, this is done by culture, which is problematic for fastidious slow-growing bacteria such as Mycobacterium avium subsp. paratuberculosis, where it can take up to 4 months to confirm growth. This study aimed to identify an assay that can rapidly quantify the number of viable M. avium subsp. paratuberculosis cells in a cellular sample. Three commercially available bacterial viability assays along with a modified liquid culture method coupled with high-throughput quantitative PCR growth detection were assessed. Criteria for assessment included the ability of each assay to differentiate live and dead M. avium subsp. paratuberculosis organisms and their accuracy at low bacterial concentrations. Using the culture-based method, M. avium subsp. paratuberculosis growth was reliably detected and quantified within 2 weeks. There was a strong linear association between the 2-week growth rate and the initial inoculum concentration. The number of viable M. avium subsp. paratuberculosis cells in an unknown sample was quantified based on the growth rate, by using growth standards. In contrast, none of the commercially available viability assays were suitable for use with samples from in vitro cellular infection assays. IMPORTANCE Rapid quantification of the viability of Mycobacterium avium subsp. paratuberculosis in samples from in vitro cellular infection assays is important, as it allows these assays to be carried out on a large scale. In vitro cellular infection assays can function as a preliminary screening tool, for vaccine development or antimicrobial screening, and also to extend findings derived from experimental animal trials. Currently, by using culture, it takes up to 4 months to obtain quantifiable results regarding M. avium subsp. paratuberculosis viability after an in vitro infection assay; however, with the quantitative PCR and liquid culture method

  14. Alginate-Encapsulation for the Improved Hypothermic Preservation of Human Adipose-Derived Stem Cells.

    Science.gov (United States)

    Swioklo, Stephen; Constantinescu, Andrei; Connon, Che J

    2016-03-01

    Despite considerable progress within the cell therapy industry, unmet bioprocessing and logistical challenges associated with the storage and distribution of cells between sites of manufacture and the clinic exist. We examined whether hypothermic (4°C-23°C) preservation of human adipose-derived stem cells could be improved through their encapsulation in 1.2% calcium alginate. Alginate encapsulation improved the recovery of viable cells after 72 hours of storage. Viable cell recovery was highly temperature-dependent, with an optimum temperature of 15°C. At this temperature, alginate encapsulation preserved the ability for recovered cells to attach to tissue culture plastic on rewarming, further increasing its effect on total cell recovery. On attachment, the cells were phenotypically normal, displayed normal growth kinetics, and maintained their capacity for trilineage differentiation. The number of cells encapsulated (up to 2 × 10(6) cells per milliliter) did not affect viable cell recovery nor did storage of encapsulated cells in a xeno-free, serum-free,current Good Manufacturing Practice-grade medium. We present a simple, low-cost system capable of enhancing the preservation of human adipose-derived stem cells stored at hypothermic temperatures, while maintaining their normal function. The storage of cells in this manner has great potential for extending the time windows for quality assurance and efficacy testing, distribution between the sites of manufacture and the clinic, and reducing the wastage associated with the limited shelf life of cells stored in their liquid state. ©AlphaMed Press.

  15. Vibrio cholerae classical biotype is converted to the viable non-culturable state when cultured with the El Tor biotype.

    Science.gov (United States)

    Pradhan, Subhra; Mallick, Sanjaya K; Chowdhury, Rukhsana

    2013-01-01

    A unique event in bacterial epidemiology was the emergence of the El Tor biotype of Vibrio cholerae O1 and the subsequent rapid displacement of the existing classical biotype as the predominant cause of epidemic cholera. We demonstrate that when the El Tor and classical biotypes were cocultured in standard laboratory medium a precipitous decline in colony forming units (CFU) of the classical biotype occurred in a contact dependent manner. Several lines of evidence including DNA release, microscopy and flow cytometric analysis indicated that the drastic reduction in CFU of the classical biotype in cocultures was not accompanied by lysis, although when the classical biotype was grown individually in monocultures, lysis of the cells occurred concomitant with decrease in CFU starting from late stationary phase. Furthermore, uptake of a membrane potential sensitive dye and protection of genomic DNA from extracellular DNase strongly suggested that the classical biotype cells in cocultures retained viability in spite of loss of culturability. These results suggest that coculturing the classical biotype with the El Tor biotype protects the former from lysis allowing the cells to remain viable in spite of the loss of culturability. The stationary phase sigma factor RpoS may have a role in the loss of culturability of the classical biotype in cocultures. Although competitive exclusion of closely related strains has been reported for several bacterial species, conversion of the target bacterial population to the viable non-culturable state has not been demonstrated previously and may have important implications in the evolution of bacterial strains.

  16. The effect of formulation on the penetration of coated and uncoated zinc oxide nanoparticles into the viable epidermis of human skin in vivo.

    Science.gov (United States)

    Leite-Silva, Vânia R; Le Lamer, Marina; Sanchez, Washington Y; Liu, David C; Sanchez, Washington H; Morrow, Isabel; Martin, Darren; Silva, Heron D T; Prow, Tarl W; Grice, Jeffrey E; Roberts, Michael S

    2013-06-01

    The use of nanoparticulate zinc oxide (ZnO-NP) in sunscreens and other cosmetic products has raised public health concerns. The two key issues are the extent of exposure to ZnO-NP and the likely hazard after the application of ZnO-NP in sunscreen and cosmetic products to humans in vivo. Our aims were to assess exposure by the extent of ZnO-NP penetration into the viable epidermis and hazard by changes in the viable epidermal redox state for a number of topical products. Of particular interest is the role of the particle coating, formulation used, and the presence of any enhancers. Multiphoton tomography with fluorescence lifetime imaging microscopy (MPT-FLIM) was used to simultaneously observe ZnO-NP penetration and potential metabolic changes within the viable epidermis of human volunteers after topical application of various ZnO-NP products. Coated and uncoated ZnO-NP remained in the superficial layers of the SC and in the skin furrows. We observed limited penetration of coated ZnO-NP dispersed in a water-in-oil emulsion formulation, which was predominantly localized adjacent to the skin furrow. However, the presence of ZnO-NP in the viable epidermis did not alter the metabolic state or morphology of the cells. In summary, our data suggest that some limited penetration of coated and uncoated ZnO-NP may occur into viable stratum granulosum epidermis adjacent to furrows, but that the extent is not sufficient to affect the redox state of those viable cells. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

  17. Growth Phase, Oxygen, Temperature and Starvation Affect the Development of Viable but Non-Culturable State of Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    Bin eWu

    2016-03-01

    Full Text Available AbstractVibrio cholerae can enter into a viable but non-culturable (VBNC state in order to survive in unfavourable environments. In this study, we studied the roles of five physicochemical and microbiological factors or states, namely, different strains, growth phases, oxygen, temperature, and starvation, on the development of VBNC of V. cholerae in artificial sea water (ASW. Different strains of the organism, the growth phase, and oxygen levels affected the progress of VBNC development. It was found that the VBNC state was induced faster in V. cholerae serogroup O1 classical biotype strain O395 than in O1 El Tor biotype strains C6706 and N16961. When cells in different growth phases were used for VBNC induction, stationary-phase cells lost their culturability more quickly than exponential-phase cells, while induction of a totally non-culturable state took longer to achieve for stationary-phase cells in all three strains, suggesting that heterogeneity of cells should be considered. Aeration strongly accelerated the loss of culturability. During the development of the VBNC state, the culturable cell count under aeration conditions was almost 106-fold lower than under oxygen-limited conditions for all three strains. The other two factors, temperature and nutrients-rich environment, may prevent the induction of VBNC cells. At 22°C or 37°C in ASW, most of the cells rapidly died and the culturable cell count reduced from about 108 CFU/mL to 106–105 CFU/mL. The total cell counts showed that cells that lost viability were decomposed, and the viable cell counts were the same as culturable cell counts, indicating that the cells did not reach the VBNC state. VBNC state development was blocked when ASW was supplied with Luria-Bertani broth (LB, but it was not affected in ASW with M9, suggesting that specific nutrients in LB may prevent the development of VBNC state. These results revealed that the five factors evaluated in this study had different

  18. Detection and Quantification of Viable and Nonviable Trypanosoma cruzi Parasites by a Propidium Monoazide Real-Time Polymerase Chain Reaction Assay

    Science.gov (United States)

    Cancino-Faure, Beatriz; Fisa, Roser; Alcover, M. Magdalena; Jimenez-Marco, Teresa; Riera, Cristina

    2016-01-01

    Molecular techniques based on real-time polymerase chain reaction (qPCR) allow the detection and quantification of DNA but are unable to distinguish between signals from dead or live cells. Because of the lack of simple techniques to differentiate between viable and nonviable cells, the aim of this study was to optimize and evaluate a straightforward test based on propidium monoazide (PMA) dye action combined with a qPCR assay (PMA-qPCR) for the selective quantification of viable/nonviable epimastigotes of Trypanosoma cruzi. PMA has the ability to penetrate the plasma membrane of dead cells and covalently cross-link to the DNA during exposure to bright visible light, thereby inhibiting PCR amplification. Different concentrations of PMA (50–200 μM) and epimastigotes of the Maracay strain of T. cruzi (1 × 105–10 parasites/mL) were assayed; viable and nonviable parasites were tested and quantified by qPCR with a TaqMan probe specific for T. cruzi. In the PMA-qPCR assay optimized at 100 μM PMA, a significant qPCR signal reduction was observed in the nonviable versus viable epimastigotes treated with PMA, with a mean signal reduction of 2.5 logarithm units and a percentage of signal reduction > 98%, in all concentrations of parasites assayed. This signal reduction was also observed when PMA-qPCR was applied to a mixture of live/dead parasites, which allowed the detection of live cells, except when the concentration of live parasites was low (10 parasites/mL). The PMA-qPCR developed allows differentiation between viable and nonviable epimastigotes of T. cruzi and could thus be a potential method of parasite viability assessment and quantification. PMID:27139452

  19. Target Contour Recovering for Tracking People in Complex Environments

    Directory of Open Access Journals (Sweden)

    Jianhua Zhang

    2012-01-01

    Full Text Available Recovering people contours from partial occlusion is a challenging problem in a visual tracking system. Partial occlusions would bring about unreasonable contour changes of the target object. In this paper, a novel method is presented to detect partial occlusion on people contours and recover occluded portions. Unlike other occlusion detection methods, the proposed method is only based on contours, which makes itself more flexible to be extended for further applications. Experiments with synthetic images demonstrate the accuracy of the method for detecting partial occlusions, and experiments on real-world video sequence are also carried out to prove that the method is also good enough to be used to recover target contours.

  20. Specifications for recovered materials. Part I. A prerequisite to marketing

    Energy Technology Data Exchange (ETDEWEB)

    An early step in planning for resource recovery, as discussed elsewhere in this issue, is marketing. But before marketing can be effectively carried out, consideration must be given to specifications for the products to be recovered; for specifications are the language between buyer and seller. Clearly, they also are important in determining the type of technology to be employed in a particular recovery facility. This is Part I of a two-part article reporting on an Environmental Protection Agency-sponsored project at the National Center for Resource Recovery to develop target specifications for materials, including a refuse-derived fuel product, recovered from municipal solid waste. Part I introduces the research and treats existing and proposed specifications for recovered paper and magnetic metals. Part II, to appear in the next issue of the NCRR Bulletin, will deal with aluminium, other non-ferrous metals, glass, and refuse-derived fuel. (MCW)

  1. Personal commentary: monitoring subsequent pregnancy in recovered peripartum cardiomyopathy mothers.

    Science.gov (United States)

    Fett, James D

    2009-12-01

    Peripartum cardiomyopathy (PPCM) is defined as the first appearance of systolic heart failure in a previously healthy woman during the last month of pregnancy or up to 6 months postpartum. Both planned and unplanned pregnancies may occur in recovered and non-recovered PPCM mothers, requiring careful counseling and management strategies. Previous studies indicate that relapse of heart failure in post-PPCM pregnancies is always a possibility, even in recovered PPCM mothers. The risk of relapse is high when the left ventricular ejection fraction is less than 0.50, and should be considered a relative contraindication to subsequent pregnancy. There are not yet established protocols for monitoring subsequent pregnancies in those who once had PPCM, and few medical articles with guidelines. This personal commentary contains parameters that have been found useful to contribute to safer monitoring with improved outcomes.

  2. Beam position monitor for energy recovered linac beams

    Science.gov (United States)

    Powers, Thomas; Evtushenko, Pavel

    2017-06-06

    A method of determining the beam position in an energy recovered linac (ERL). The method makes use of in phase and quadrature (I/Q) demodulation techniques to separate the pickup signal generated by the electromagnetic fields generated by the first and second pass beam in the energy recovered linac. The method includes using analog or digital based I/Q demodulation techniques in order to measure the relative amplitude of the signals from a position sensitive beam pickup such as a button, strip line or microstripline beam position monitor.

  3. RECYCLING RECOVERED WOOD-BASED MATERIALS. FLATNESS, MOR AND MOE OF PANELS MADE FROM RECOVERED WOODEN BASED MATERIALS

    Directory of Open Access Journals (Sweden)

    Andrea DEÁK

    2015-12-01

    Full Text Available The objective of the research was to study the mechanical properties of recovered solid wood and wood-based materials, to create new panels made of strips of recovered oak and spruce wood, particleboards and blockboards and to study their mechanical properties, feasibility and use. Flatness, MOR and MOE were determined for the solid wood and wood-based materials recovered from construction and demolition sites. Four types of experimental panels (PL1, PL2, PALL1 and PALL2 were manufactured from solid wood strips alternating with either particleboard or blockboard strips glued edge to edge. Some of their physical and mechanical properties were investigated. All new panels made from recovered materials fell within allowable limits specified in European standard requirements for panel flatness. Panels type PL2, PALL1 and PALL2 show better values for MoR and MoE parallel to the grain when compared to the respective values for the recovered wood-based materials of which they are made, but panels type PL1 show no change. Panels type PALL2 are preferable to panels type PALL1 due to better mechanical properties. Panels type PL1 and PL2 show no such differences in their structure. The results also show that these new panels can be used successfully in furniture production, provided panels type PL1, PL2 ad PALL1 do not bear loads perpendicular to the grain or receive reinforcing elements.

  4. Comparison of spike and aerosol challenge tests for the recovery of viable influenza virus from non-woven fabrics.

    Science.gov (United States)

    Zuo, Zhili; de Abin, Martha; Chander, Yogesh; Kuehn, Thomas H; Goyal, Sagar M; Pui, David Y H

    2013-09-01

    To experimentally determine the survival kinetics of influenza virus on personal protective equipment (PPE) and to evaluate the risk of virus transfer from PPE, it is important to compare the effects on virus recovery of the method used to contaminate the PPE with virus and the type of eluent used to recover it. Avian influenza virus (AIV) was applied as a liquid suspension (spike test) and as an aerosol to three types of non-woven fabrics [polypropylene (PP), polyester (PET), and polyamide (Nylon)] that are commonly used in the manufacture of PPE. This was followed by virus recovery using eight different eluents (phosphate-buffered saline, minimum essential medium, and 1.5% or 3.0% beef extract at pH 7, 8, or 9). For spike tests, no statistically significant difference was found in virus recovery using any of the eluents tested. Hydrophobic surfaces (PP and PET) yielded higher spiked virus recovery than hydrophilic Nylon. From all materials, the virus recovery was much lower in aerosol challenge tests than in spike tests. Significant differences were found in the recovery of viable AIV from non-woven fabrics between spike and aerosol challenge tests. The findings of this study demonstrate the need for realistic aerosol challenge tests rather than liquid spike tests in studies of virus survival on surfaces where airborne transmission of influenza virus may get involved. © 2013 John Wiley & Sons Ltd.

  5. Detection of viable Helicobacter pylori inside free-living amoebae in wastewater and drinking water samples from Eastern Spain.

    Science.gov (United States)

    Moreno-Mesonero, Laura; Moreno, Yolanda; Alonso, José Luis; Ferrús, M Antonia

    2017-10-01

    Helicobacter pylori is one of the most concerning emerging waterborne pathogens. It has been suggested that it could survive in water inside free-living amoebae (FLA), but nobody has studied this relationship in the environment yet. Thus, we aimed to detect viable H. pylori cells from inside FLA in water samples. Sixty-nine wastewater and 31 drinking water samples were collected. FLA were purified and identified by PCR and sequencing. For exclusively detecting H. pylori inside FLA, samples were exposed to sodium hypochlorite and assayed by specific PMA-qPCR, DVC-FISH and culture. FLA were detected in 38.7% of drinking water and 79.7% of wastewater samples, even after disinfection. In wastewater, Acanthamoeba spp. and members of the family Vahlkampfiidae were identified. In drinking water, Acanthamoeba spp. and Echinamoeba and/or Vermamoeba were present. In 39 (58.2%) FLA-positive samples, H. pylori was detected by PMA-qPCR. After DVC-FISH, 21 (31.3%) samples harboured viable H. pylori internalized cells. H. pylori was cultured from 10 wastewater samples. To our knowledge, this is the first report that demonstrates that H. pylori can survive inside FLA in drinking water and wastewater, strongly supporting the hypothesis that FLA could play an important role in the transmission of H. pylori to humans. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  6. Escherichia coli detection using mTEC agar and fluorescent antibody direct viable counting on coastal recreational water samples.

    Science.gov (United States)

    Zimmerman, A M; Rebarchik, D M; Flowers, A R; Williams, J L; Grimes, D J

    2009-10-01

    Escherichia coli is the faecal indicator species recommended by the US Environmental Protection Agency (USEPA) for monitoring fresh recreational water. Viable but nonculturable (VBNC) E. coli are living cells that are dormant and not culturable using standard microbiological cultivation methods. This study reports a comparison between the mTEC culture method recommended by USEPA for E. coli enumeration and a fluorescent antibody-direct viable count (FA-DVC) method to visualize living E. coli cells with a microscope. Escherichia coli, faecal coliforms and Enterococcus were detected using standard methods recommended by the USEPA. VBNC E. coli was visualized with FA-DVC. Results were analysed with standard statistical methods (Pearson correlation; paired-sample t-test). Significantly higher numbers of E. coli were detected using the FA-DVC method than using the mTEC method. Escherichia coli results were also compared with faecal coliform (mFC broth) and Enterococcus (mEI agar) counts in the same samples. The results of this comparative study demonstrate that E. coli can be present in higher numbers than what are detected with standard culture methods. This study re-emphasizes the need for a rapid, accurate and precise method for detecting health risks to humans who use recreational waters.

  7. Isolation of viable human hepatic progenitors from adult livers is possible even after 48 hours of cold ischemia.

    Science.gov (United States)

    Aupet, Sophie; Simoné, Gael; Heyd, Bruno; Bachellier, Philippe; Vidal, Isabelle; Richert, Lysiane; Martin, Hélène

    2013-07-01

    Liver transplantation, utilized routinely for end-stage liver disease, has been constrained by the paucity of organ donors, and is being complemented by alternative strategies such as liver cell transplantation. One of the most promising forms of liver cell transplantation is hepatic stem cell therapies, as the number of human hepatic stem cells (hHpSCs) and other early hepatic progenitor cells (HPCs) are sufficient to provide treatment for multiple patients from a single liver source. In the present study, human adult livers were exposed to cold ischemia and then processed after numbers, albeit somewhat lower, were obtained from those exposed to 48 h of cold ischemia. The yields are similar to those reported from livers with minimal exposure to ischemia. When cultured on plastic dishes and in Kubota's Medium, a serum-free medium designed for early lineage stage HPCs, colonies of rapidly expanding cells formed. They were confirmed to be probable hHpSCs by their ability to survive and expand on plastic and in Kubota's Medium for months, by co-expression of EpCAM and neural cell adhesion molecule, minimal if any albumin expression, with EpCAM found throughout the cells, and no expression of alpha-fetoprotein. The yields of viable EpCAM(+) cells were surprisingly large, and the numbers from a single donor liver are sufficient to treat approximately 50-100 patients given the numbers of EpCAM(+) cells currently used in hepatic stem cell therapies. Thus, cold ischemic livers for up to 48 h are a new source of cells that might be used for liver cell therapies.

  8. Clinically Viable Gene Expression Assays with Potential for Predicting Benefit from MEK Inhibitors.

    Science.gov (United States)

    Brant, Roz; Sharpe, Alan; Liptrot, Tom; Dry, Jonathan R; Harrington, Elizabeth A; Barrett, J Carl; Whalley, Nicky; Womack, Christopher; Smith, Paul; Hodgson, Darren R

    2017-03-15

    Purpose: To develop a clinically viable gene expression assay to measure RAS/RAF/MEK/ERK (RAS-ERK) pathway output suitable for hypothesis testing in non-small cell lung cancer (NSCLC) clinical studies. Experimental Design: A published MEK functional activation signature (MEK signature) that measures RAS-ERK functional output was optimized for NSCLC in silico NanoString assays were developed for the NSCLC optimized MEK signature and the 147-gene RAS signature. First, platform transfer from Affymetrix to NanoString, and signature modulation following treatment with KRAS siRNA and MEK inhibitor, were investigated in cell lines. Second, the association of the signatures with KRAS mutation status, dynamic range, technical reproducibility, and spatial and temporal variation was investigated in NSCLC formalin-fixed paraffin-embedded tissue (FFPET) samples. Results: We observed a strong cross-platform correlation and modulation of signatures in vitro Technical and biological replicates showed consistent signature scores that were robust to variation in input total RNA; conservation of scores between primary and metastatic tumor was statistically significant. There were statistically significant associations between high MEK ( P = 0.028) and RAS ( P = 0.003) signature scores and KRAS mutation in 50 NSCLC samples. The signatures identify overlapping but distinct candidate patient populations from each other and from KRAS mutation testing. Conclusions: We developed a technically and biologically robust NanoString gene expression assay of MEK pathway output, compatible with the quantities of FFPET routinely available. The gene signatures identified a different patient population for MEK inhibitor treatment compared with KRAS mutation testing. The predictive power of the MEK signature should be studied further in clinical trials. Clin Cancer Res; 23(6); 1471-80. ©2016 AACR See related commentary by Xue and Lito, p. 1365 . ©2016 American Association for Cancer Research.

  9. Immediate natural tooth pontic: A viable yet temporary prosthetic solution: A patient reported outcome

    Directory of Open Access Journals (Sweden)

    Sudhir Bhandari

    2012-01-01

    Conclusion: The concept of immediate pontic placement is surely a viable treatment option and promises an excellent transient esthetic solution for a lost tooth as well as enables good preparation of the extraction site for future prosthetic replacement.

  10. Improved identification of viable myocardium using second harmonic imaging during dobutamine stress echocardiography

    NARCIS (Netherlands)

    F. Sozzi (Fabiola); D. Poldermans (Don); J.J. Bax (Jeroen); A. Elhendy (Abdou); E.C. Vourvouri (Eleni); R. Valkema (Roelf); J. de Sutter; A.F.L. Schinkel (Arend); A. Borghetti; J.R.T.C. Roelandt (Jos)

    2001-01-01

    textabstractOBJECTIVE: To determine whether, compared with fundamental imaging, second harmonic imaging can improve the accuracy of dobutamine stress echocardiography for identifying viable myocardium, using nuclear imaging as a reference. PATIENTS: 30 patients with chronic left

  11. Marine environmental pollution stress detection through direct viable counts of bacteria

    Digital Repository Service at National Institute of Oceanography (India)

    Ramaiah, N.; Kenkre, V.D.; Verlecar, X.N.

    Direct viable counts (DVC) of bacteria were quantified from polluted and relatively less/non-polluted coastal locations during different seasons to assess whether they can be routinely monitored for an understanding of environmental stress(es...

  12. Recovering sulfurs from wastes; Recuperacion de sulfuros desde efluentes

    Energy Technology Data Exchange (ETDEWEB)

    Schneider, A.; Guala, M.; Flores, H. [Universidad Nacional del Litoral (UNL), Santa Fe (Argentina). Facultad de Ingenieria Quimica; Spekuljak, Z; Robaina, E. [Consejo Nacional de Investigaciones Cientificas y Tecnologicas (CONICET), Buenos Aires (Argentina)

    2000-07-01

    A tannery sulfur recuperation process is proposed. The effluent liquid is supersaturated with sulphydric acid, by means acidification at controlled temperature and pressure, and a consequent venting of the clean gas. It is absorbed in a basic solution, and recovering for reutilization. The lab experiences and the results are presented. Technological solution are found to employ this process in industrial applications. (author)

  13. Colonizing and Decolonizing Projects of Re/covering Spirituality

    Science.gov (United States)

    Rhee, Jeong-eun; Subedi, Binaya

    2014-01-01

    In this postcolonial inquiry, we analyze how spirituality has been simultaneously appropriated/re-covered and re-appropriated/recovered for the purpose of (re)colonizing as well as decolonizing projects. By drawing from discrete yet interconnected literatures of decolonizing, (post)(anti)colonial, Indigenous, and ethnic studies based theories, we…

  14. Evaluation of an automated struvite reactor to recover phosphorus ...

    African Journals Online (AJOL)

    2015-04-03

    Apr 3, 2015 ... urine is an ideal source for fertilizer production, as the majority of nutrients found in wastewater streams originate from urine. (Larsen et al., 1996). One way to extract nutrients from stored urine is by adding soluble magnesium to recover phosphate by precipitating struvite (magnesium ammonium phosphate ...

  15. 33 CFR 385.20 - Restoration Coordination and Verification (RECOVER).

    Science.gov (United States)

    2010-07-01

    .... (b) Documents or work products prepared or developed by RECOVER shall not be self-executing, but... adaptive management program; (iii) Conducting monitoring and assessment activities as part of the adaptive... conduct planning and integration activities, in accordance with § 385.31, in support of the adaptive...

  16. False or Recovered Memories?: Legal and Ethical Implications for Therapists.

    Science.gov (United States)

    Jenkins, Peter

    1997-01-01

    Places the development of the debate over false or recovered memories in its social and historical context. Identifies some of the ethical and legal implications of this area of work for therapists by using the Drama Triangle. Outlines ethical dilemmas for therapists and some of the implications for therapeutic practice. (RJM)

  17. Beyond coherence: Recovering structured time-frequency representations

    DEFF Research Database (Denmark)

    Borup, Lasse; Gribonval, Rémi; Nielsen, Morten

    2008-01-01

    We consider the problem of recovering a structured sparse representation of a signal in an overcomplete time-frequency dictionary with a particular structure. For infinite dictionaries that are the union of a nice wavelet basis and a Wilson basis, sufficient conditions are given for the basis...

  18. Beyond coherence: Recovering structured time-frequency represenatations

    DEFF Research Database (Denmark)

    Borup, Lasse; Gribonval, Rémi; Nielsen, Morten

    We consider the problem of recovering a structured sparse representation of a signal in an overcomplete time-frequency dictionary with a particular structure. For infinite dictionaries that are the union of a nice wavelet basis and a Wilson basis, sufficient conditions are given for the Basis...

  19. Behandling kan hjelpe tareskogen : Treatment may help kelp to recover

    OpenAIRE

    Sunnset, Beate Hoddevik

    2010-01-01

    After successful experiments to remove sea urchins with quicklime, the kelp forest is returning to the test areas in Porsangerfjorden in Finnmark. Juvenile fish appear to be thriving amongst the new kelp plants, which makes us optimistic that both the kelp forests and fish populations in the fjord can recover

  20. Blessing of Dimensionality: Recovering Mixture Data via Dictionary Pursuit.

    Science.gov (United States)

    Liu, Guangcan; Liu, Qingshan; Li, Ping

    2017-01-01

    This paper studies the problem of recovering the authentic samples that lie on a union of multiple subspaces from their corrupted observations. Due to the high-dimensional and massive nature of today's data-driven community, it is arguable that the target matrix (i.e., authentic sample matrix) to recover is often low-rank. In this case, the recently established Robust Principal Component Analysis (RPCA) method already provides us a convenient way to solve the problem of recovering mixture data. However, in general, RPCA is not good enough because the incoherent condition assumed by RPCA is not so consistent with the mixture structure of multiple subspaces. Namely, when the subspace number grows, the row-coherence of data keeps heightening and, accordingly, RPCA degrades. To overcome the challenges arising from mixture data, we suggest to consider LRR in this paper. We elucidate that LRR can well handle mixture data, as long as its dictionary is configured appropriately. More precisely, we mathematically prove that LRR can weaken the dependence on the row-coherence, provided that the dictionary is well-conditioned and has a rank of not too high. In particular, if the dictionary itself is sufficiently low-rank, then the dependence on the row-coherence can be completely removed. These provide some elementary principles for dictionary learning and naturally lead to a practical algorithm for recovering mixture data. Our experiments on randomly generated matrices and real motion sequences show promising results.

  1. Sensitivity of Listeria species, recovered from indigenous chickens ...

    African Journals Online (AJOL)

    Background: Resistance of bacteria to antibiotics and disinfectants has been reported widely in the world. Listeria monocytogenes is no exception, although normally it tends to be variably sensitive to many antibiotics and disinfectants. Objectives: To assess the susceptibility of Listeria isolates recovered from indigenous ...

  2. The value of DNA material recovered from crime scenes.

    Science.gov (United States)

    Bond, John W; Hammond, Christine

    2008-07-01

    DNA material is now collected routinely from crime scenes for a wide range of offenses and its timely processing is acknowledged as a key element to its success in solving crime. An analysis of the processing of approximately 1500 samples of DNA material recovered from the property crime offenses of residential burglary, commercial burglary, and theft of motor vehicle in Northamptonshire, U.K. during 2006 identified saliva and cigarette ends as the main sources of DNA recovered (approximately 63% of samples) with blood, cellular DNA, and chewing gum accounting for the remainder. The conversion of these DNA samples into DNA profiles and then into matches with offender profiles held on the U.K. National DNA database is considered in terms of the ease with which Crime Scene Examiners can recover DNA rich samples of different sources, the location of the DNA at the crime scene, and its mobility. A logistical regression of the DNA material recovered has revealed a number of predictors, other than timeliness, that greatly influence its conversion into a DNA profile. The most significant predictor was found to be Crime Scene Examiner accreditation with offense type and DNA sample condition also being relevant. A similar logistical regression of DNA samples profiled that produced a match with an offender on the U.K. National DNA database showed no significance with any of the predictors considered.

  3. Return to Education for Recovering Drug Addicts: The Soilse Project

    Science.gov (United States)

    Barter, Eric

    2010-01-01

    This article is an account of a return to education course set up to cater to the needs of recovering heroin addicts in a Dublin rehabilitation project in the summer of 2008. It begins with a brief outline of the HSE Soilse rehabilitation and recovery programme and the rationale for seeking association with the Department of Adult and Community…

  4. Determination of Shooting Distance from Deformation of the Recovered Bullet

    Science.gov (United States)

    1987-01-01

    shooting distance was over 100 yds (90 in). Personnel at the State Criminalistics Laboratory were aware of work done at the Wound Ballistics Laboratory of...ies 1.31. The query by the California Department of Justice Criminalistic Laboratory concerning the determination of shooting distance from a recovered

  5. ARGUMENTS FOR REUSING OLD OAK WOOD RECOVERED FROM DEMOLITION

    Directory of Open Access Journals (Sweden)

    Andrea DEÁK

    2015-09-01

    Full Text Available The main objective of this study was to find scientific arguments in favour of reusing recovered oak wood in order to reintegrate it in new types of structures. For this study six beams were selected, which had been recovered from a house built in 1831. Visual analysis revealed several defects and signs of degradation in the sapwood and inside the cracks. The recovered beams hadn’t been treated with any toxic chemicals. Bending strength, modulus of elasticity and compression strength parallel to the grain were investigated and microscopy and FTIR-ATR spectroscopy were used to check the quality of the apparently sound beam areas also used for the determination of mechanical properties. Macroscopic and microscopic analysis may not reveal any incipient fungal attack but FTIR analysis was proven to be effective. The obtained results open a new perspective for the future use of recovered old oak wood, due to the adequate mechanical properties and to the assumed means of eliminating degraded areas.

  6. A probabilistic model to recover individual genomes from metagenomes

    NARCIS (Netherlands)

    J. Dröge (Johannes); A. Schönhuth (Alexander); A.C. McHardy (Alice)

    2017-01-01

    textabstractShotgun metagenomics of microbial communities reveal information about strains of relevance for applications in medicine, biotechnology and ecology. Recovering their genomes is a crucial but very challenging step due to the complexity of the underlying biological system and technical

  7. Phosphorus uptake by potato from fertilizers recovered from anaerobic digestion

    Science.gov (United States)

    Field experiments were conducted in the Columbia Basin of South Central Washington to assess the yield of potato (Solanum tuberosum) in response to application of phosphorus enriched materials recovered from anaerobic digestion of manure. The treatments were comprised of four rates (0, 56, 112 and ...

  8. Thermoset composite recycling: Properties of recovered glass fiber

    DEFF Research Database (Denmark)

    Beauson, Justine; Fraisse, Anthony; Toncelli, C.

    2015-01-01

    Recycling of glass fiber thermoset polymer composite is a challenging topic and a process able to recover the glass fibers original properties in a limited cost is still under investigation. This paper focuses on the recycling technique separating the glass fiber from the matrix material. Four...

  9. Physical activity level among children recovering from severe acute malnutrition

    DEFF Research Database (Denmark)

    Babirekere-Iriso, Esther; Rytter, Maren Johanne Heilskov; Namusoke, Hanifa

    2018-01-01

    OBJECTIVE: To assess the level and predictors of physical activity at discharge among children recovering from severe acute malnutrition (SAM). METHODS: We conducted a prospective study among 69 children 6 - 59 months of age admitted with SAM for nutritional rehabilitation at Mulago National...

  10. Immunization of rodents against Hymenolepis infections using non-viable homologous oncospheres.

    Science.gov (United States)

    Fan, Ping-Chin; Chung, Wen-Cheng; Ito, Akira

    2004-12-01

    Immunity to Taiwan Taenia infection in pigs can be stimulated using homologous or heterologous non-viable Taenia oncospheres. This study was designed to determine whether homologous non-viable oncospheres could stimulate immunity to Hymenolepis infection in rodents. Hatched oncospheres were prepared from eggs of Hymenolepis diminuta, Hymenolepis nana, and Hymenolepis microstoma and kept at -70 degrees C for more than 1 month. A mixture of 500 non-viable oncospheres of each tapeworm and complete Freund's adjuvant was injected subcutaneously in four groups of Sprague-Dawley rats or ICR mice one to four times at an interval of 1 week; controls were not immunized. After immunization, each rodent was orally inoculated with three fresh active cysticercoids of H. diminuta or H. microstoma or 500 fresh eggs of H. nana. The animals were then necropsied for adult tapeworms. No rats or mice immunized with non-viable oncospheres of H. diminuta or H. nana were infected by the challenge inoculation. However, 28 of 34 mice immunized with non-viable H. microstoma oncospheres were infected after inoculation with cysticercoids. This study demonstrated complete protection against infection by homologous parasites in rats or mice immunized with non-viable oncospheres of H. diminuta and H. nana, respectively. Repeated immunization may not be required if resistance is stimulated in rodent hosts.

  11. [Viable myocardium detecting by CARTO voltage mapping in swine model of acute myocardial infarction].

    Science.gov (United States)

    Lin, Tao; Ma, Yi-Tong; Yang, Yi-Ning; Mu, Hu-Yati; He, Peng-Yi; Yang, Yu-Chun; Chou, Ping; Liu, Fen; Zhang, Yan-Yi

    2010-08-01

    To evaluate the accuracy and practicability of detecting viable myocardium by CARTO voltage mapping in swine model of acute myocardial infarction (MI). MI was induced in 13 anesthetized swines via occluding the distal of left anterior descending coronary arteries by angioplasty balloon for 60-90 minutes. The viable myocardium detection by CARTO voltage mapping was made after reconstruction of the left ventricle using CARTO and the results were compared with TTC staining. The standard of CARTO voltage to detect viable myocardium was 0.5 - 1.5 mV while viable myocardium showed pink color by TTC staining. Eleven out of 13 swines survived the operation and 2 swines died of ventricular fibrillation at 45 and 65 minutes post ischemia. Left ventricle was divided into 16 segments and 176 segments from 11 swines were analyzed. Viable myocardium detected by CARTO voltage mapping was identical as identified by TTC staining (Kappa = 0.816, P < 0.001). Taken the TTC result as standard, the sensitivity, specificity and accuracy rate of CARTO voltage mapping are 71.8%, 96.5% and 90.9% respectively. CARTO voltage mapping could be used as a reliable tool to detect viable myocardium in this model.

  12. Characterization of nonpolio enteroviruses recovered from patients with aseptic meningitis in Korea.

    Science.gov (United States)

    Joo, Chul Hyun; Ahn, Jeonghyun; Seo, Ilsun; Kim, Yoo Kyum; Kim, Donghou; Hong, Heanam; Lee, Heuiran

    2005-01-01

    We attempted to characterize nonpolio enteroviruses recovered from Korean patients with aseptic meningitis. We performed RT-PCR on the 5'-nontranslated region using clinical specimens. Infectious clinical isolates were amplified by infecting Vero cells with RT-PCR-positive clinical specimens. We then investigated the direct effect in primary neuronal cells or cardiomyocytes following virus infection. Total 12 clinical isolates were subtypically analyzed by both RT-PCR/sequencing comparison of the VP-1 region and neutralization assay. 43-2, 43-2S, 57 and 58 were found to be coxsackievirus B1 (CVB1), 312 to be CVB5, 14-2S and 327 to be echovirus 6, 165 to be echovirus 9, 337 to be echovirus 11, and 270 to be echovirus 30. All the clinical isolates tested showed profound cytotoxicity to various degrees in the primary neuronal cells within 24 h postinfection at 10 MOI. By contrast, a significant cytopathic effect was observed in the primary cardiomyocytes at 3-5 days postinfection at 50 MOI. The present study suggests that the clinical isolates recovered from Korean patients belonged to different CVB or echovirus serotypes and that these viruses showed diversities in their virulence in primary neuronal cells and cardiomyocytes. Copyright (c) 2005 S. Karger AG, Basel.

  13. “Limits of Control” – Crucial Parameters for a Reliable Quantification of Viable Campylobacter by Real-Time PCR

    Science.gov (United States)

    Krüger, Nora-Johanna; Buhler, Christiane; Iwobi, Azuka N.; Huber, Ingrid; Ellerbroek, Lüppo; Appel, Bernd; Stingl, Kerstin

    2014-01-01

    The unsuitability of the “CFU” parameter and the usefulness of cultivation-independent quantification of Campylobacter on chicken products, reflecting the actual risk for infection, is increasingly becoming obvious. Recently, real-time PCR methods in combination with the use of DNA intercalators, which block DNA amplification from dead bacteria, have seen wide application. However, much confusion exists in the correct interpretation of such assays. Campylobacter is confronted by oxidative and cold stress outside the intestine. Hence, damage caused by oxidative stress probably represents the most frequent natural death of Campylobacter on food products. Treatment of Campylobacter with peroxide led to complete loss of CFU and to significant entry of any tested DNA intercalator, indicating disruption of membrane integrity. When we transiently altered the metabolic state of Campylobacter by abolishing the proton-motive force or by inhibiting active efflux, CFU was constant but enhanced entry of ethidium bromide (EtBr) was observed. Consistently, ethidium monoazide (EMA) also entered viable Campylobacter, in particular when nutrients for bacterial energization were lacking (in PBS) or when the cells were less metabolically active (in stationary phase). In contrast, propidium iodide (PI) and propidium monoazide (PMA) were excluded from viable bacterial cells, irrespective of their metabolic state. As expected for a diffusion-limited process, the extent of signal reduction from dead cells depended on the temperature, incubation time and concentration of the dyes during staining, prior to crosslinking. Consistently, free protein and/or DNA present in varying amounts in the heterogeneous matrix lowered the concentration of the DNA dyes at the bacterial membrane and led to considerable variation of the residual signal from dead cells. In conclusion, we propose an improved approach, taking into account principles of method variability and recommend the implementation of

  14. "Limits of control"--crucial parameters for a reliable quantification of viable campylobacter by real-time PCR.

    Science.gov (United States)

    Krüger, Nora-Johanna; Buhler, Christiane; Iwobi, Azuka N; Huber, Ingrid; Ellerbroek, Lüppo; Appel, Bernd; Stingl, Kerstin

    2014-01-01

    The unsuitability of the "CFU" parameter and the usefulness of cultivation-independent quantification of Campylobacter on chicken products, reflecting the actual risk for infection, is increasingly becoming obvious. Recently, real-time PCR methods in combination with the use of DNA intercalators, which block DNA amplification from dead bacteria, have seen wide application. However, much confusion exists in the correct interpretation of such assays. Campylobacter is confronted by oxidative and cold stress outside the intestine. Hence, damage caused by oxidative stress probably represents the most frequent natural death of Campylobacter on food products. Treatment of Campylobacter with peroxide led to complete loss of CFU and to significant entry of any tested DNA intercalator, indicating disruption of membrane integrity. When we transiently altered the metabolic state of Campylobacter by abolishing the proton-motive force or by inhibiting active efflux, CFU was constant but enhanced entry of ethidium bromide (EtBr) was observed. Consistently, ethidium monoazide (EMA) also entered viable Campylobacter, in particular when nutrients for bacterial energization were lacking (in PBS) or when the cells were less metabolically active (in stationary phase). In contrast, propidium iodide (PI) and propidium monoazide (PMA) were excluded from viable bacterial cells, irrespective of their metabolic state. As expected for a diffusion-limited process, the extent of signal reduction from dead cells depended on the temperature, incubation time and concentration of the dyes during staining, prior to crosslinking. Consistently, free protein and/or DNA present in varying amounts in the heterogeneous matrix lowered the concentration of the DNA dyes at the bacterial membrane and led to considerable variation of the residual signal from dead cells. In conclusion, we propose an improved approach, taking into account principles of method variability and recommend the implementation of

  15. "Limits of control"--crucial parameters for a reliable quantification of viable campylobacter by real-time PCR.

    Directory of Open Access Journals (Sweden)

    Nora-Johanna Krüger

    Full Text Available The unsuitability of the "CFU" parameter and the usefulness of cultivation-independent quantification of Campylobacter on chicken products, reflecting the actual risk for infection, is increasingly becoming obvious. Recently, real-time PCR methods in combination with the use of DNA intercalators, which block DNA amplification from dead bacteria, have seen wide application. However, much confusion exists in the correct interpretation of such assays. Campylobacter is confronted by oxidative and cold stress outside the intestine. Hence, damage caused by oxidative stress probably represents the most frequent natural death of Campylobacter on food products. Treatment of Campylobacter with peroxide led to complete loss of CFU and to significant entry of any tested DNA intercalator, indicating disruption of membrane integrity. When we transiently altered the metabolic state of Campylobacter by abolishing the proton-motive force or by inhibiting active efflux, CFU was constant but enhanced entry of ethidium bromide (EtBr was observed. Consistently, ethidium monoazide (EMA also entered viable Campylobacter, in particular when nutrients for bacterial energization were lacking (in PBS or when the cells were less metabolically active (in stationary phase. In contrast, propidium iodide (PI and propidium monoazide (PMA were excluded from viable bacterial cells, irrespective of their metabolic state. As expected for a diffusion-limited process, the extent of signal reduction from dead cells depended on the temperature, incubation time and concentration of the dyes during staining, prior to crosslinking. Consistently, free protein and/or DNA present in varying amounts in the heterogeneous matrix lowered the concentration of the DNA dyes at the bacterial membrane and led to considerable variation of the residual signal from dead cells. In conclusion, we propose an improved approach, taking into account principles of method variability and recommend the

  16. Evaluation of propidium monoazide real-time PCR for early detection of viable Mycobacterium tuberculosis in clinical respiratory specimens.

    Science.gov (United States)

    Kim, Young Jin; Lee, Sun Min; Park, Byung Kyu; Kim, Sung Soo; Yi, Jongyoun; Kim, Hyung Hoi; Lee, Eun Yup; Chang, Chulhun Ludgerus

    2014-05-01

    Conventional acid-fast bacilli (AFB) staining cannot differentiate viable from dead cells. Propidium monoazide (PMA) is a photoreactive DNA-binding dye that inhibits PCR amplification by DNA modification. We evaluated whether PMA real-time PCR is suitable for the early detection of viable Mycobacterium tuberculosis (MTB) in clinical respiratory specimens. A total of 15 diluted suspensions from 5 clinical MTB isolates were quadruplicated and subjected to PMA treatment and/or heat inactivation. Eighty-three AFB-positive sputum samples were also tested to compare the ΔCT values (CT value in PMA-treated sputum samples-CT value in non-PMA-treated sputum samples) between culture-positive and culture-negative specimens. Real-time PCR was performed using Anyplex MTB/NTM Real-Time Detection (Seegene, Korea), and the CT value changes after PMA treatment were compared between culture-positive and culture-negative groups. In MTB suspensions, the increase in the CT value after PMA treatment was significant in dead cells (P=0.0001) but not in live cells (P=0.1070). In 14 culture-negative sputum samples, the median ΔCT value was 5.3 (95% confidence interval [CI], 4.1-8.2; P<0.0001), whereas that in 69 culture-positive sputum samples was 1.1 (95% CI, 0.7-2.0). In the ROC curve analysis, the cutoff ΔCT value for maximum sensitivity (89.9%) and specificity (85.7%) for differentiating dead from live cells was 3.4. PMA real-time PCR is a useful approach for differentiating dead from live bacilli in AFB smear-positive sputum samples.

  17. Detection of viable Salmonella in ice cream by TaqMan real-time polymerase chain reaction assay combining propidium monoazide.

    Science.gov (United States)

    Wang, Yuexia; Yang, Ming; Liu, Shuchun; Chen, Wanyi; Suo, Biao

    2015-09-01

    Real-time polymerase chain reaction (PCR) allows rapid detection of Salmonella in frozen dairy products, but it might cause a false positive detection result because it might amplify DNA from dead target cells as well. In this study, Salmonella-free frozen ice cream was initially inoculated with heat-killed Salmonella Typhimurium cells and stored at -18°C. Bacterial DNA extracted from the sample was amplified using TaqMan probe-based real-time PCR targeting the invA gene. Our results indicated that DNA from the dead cells remained stable in frozen ice cream for at least 20 days, and could produce fluorescence signal for real-time PCR as well. To overcome this limitation, propidium monoazide (PMA) was combined with real-time PCR. PMA treatment can effectively prevent PCR amplification from heat-killed Salmonella cells in frozen ice cream. The PMA real-time PCR assay can selectively detect viable Salmonella at as low as 103 CFU/mL. Combining 18 hours of pre-enrichment with the assay allows for the detection of viable Salmonella at 100 CFU/mL and avoiding the false-positive result of dead cells. The PMA real-time PCR assay provides an alternative specifically for detection of viable Salmonella in ice cream. However, when the PMA real-time PCR assay was evaluated in ice cream subjected to frozen storage, it obviously underestimated the contamination situation of viable Salmonella, which might lead to a false negative result. According to this result, the use of enrichment prior to PMA real-time PCR analysis remains as the more appropriate approach. Copyright © 2015. Published by Elsevier B.V.

  18. Detection of viable Salmonella in ice cream by TaqMan real-time polymerase chain reaction assay combining propidium monoazide

    Directory of Open Access Journals (Sweden)

    Yuexia Wang

    2015-09-01

    Full Text Available Real-time polymerase chain reaction (PCR allows rapid detection of Salmonella in frozen dairy products, but it might cause a false positive detection result because it might amplify DNA from dead target cells as well. In this study, Salmonella-free frozen ice cream was initially inoculated with heat-killed Salmonella Typhimurium cells and stored at −18°C. Bacterial DNA extracted from the sample was amplified using TaqMan probe-based real-time PCR targeting the invA gene. Our results indicated that DNA from the dead cells remained stable in frozen ice cream for at least 20 days, and could produce fluorescence signal for real-time PCR as well. To overcome this limitation, propidium monoazide (PMA was combined with real-time PCR. PMA treatment can effectively prevent PCR amplification from heat-killed Salmonella cells in frozen ice cream. The PMA real-time PCR assay can selectively detect viable Salmonella at as low as 103 CFU/mL. Combining 18 hours of pre-enrichment with the assay allows for the detection of viable Salmonella at 100 CFU/mL and avoiding the false-positive result of dead cells. The PMA real-time PCR assay provides an alternative specifically for detection of viable Salmonella in ice cream. However, when the PMA real-time PCR assay was evaluated in ice cream subjected to frozen storage, it obviously underestimated the contamination situation of viable Salmonella, which might lead to a false negative result. According to this result, the use of enrichment prior to PMA real-time PCR analysis remains as the more appropriate approach.

  19. Assessments of total and viable Escherichia coli O157:H7 on field and laboratory grown lettuce.

    Directory of Open Access Journals (Sweden)

    Anne-Laure Moyne

    Full Text Available Leafy green produce has been associated with numerous outbreaks of foodborne illness caused by strains of Escherichia coli O157:H7. While the amounts of culturable E. coli O157:H7 rapidly decline after introduction onto lettuce in the field, it remains to be determined whether the reduction in cell numbers is due to losses in cell viability, cell injury and a subsequent inability to be detected by standard laboratory culturing methods, or a lack of adherence and hence rapid removal of the organism from the plants during application. To assess which of these options is most relevant for E. coli O157:H7 on leafy green produce, we developed and applied a propidium monoazide (PMA real-time PCR assay to quantify viable (with PMA and total (without PMA E. coli O157:H7 cells on growth chamber and field-grown lettuce. E. coli O157:H7, suspended in 0.1% peptone, was inoculated onto 4-week-old lettuce plants at a level of approximately 10(6 CFU/plant. In the growth chamber at low relative humidity (30%, culturable amounts of the nontoxigenic E. coli O157:H7 strain ATCC 700728 and the virulent strain EC4045 declined 100 to 1000-fold in 24 h. Fewer E. coli O157:H7 cells survived when applied onto plants in droplets with a pipette compared with a fine spray inoculation. Total cells for both strains were equivalent to inoculum levels for 7 days after application, and viable cell quantities determined by PMA real-time PCR were approximately 10(4 greater than found by colony enumeration. Within 2 h after application onto plants in the field, the number of culturable E. coli ATCC 700728 was reduced by up to 1000-fold, whereas PCR-based assessments showed that total cell amounts were equivalent to inoculum levels. These findings show that shortly after inoculation onto plants, the majority of E. coli O157:H7 cells either die or are no longer culturable.

  20. Assessments of Total and Viable Escherichia coli O157:H7 on Field and Laboratory Grown Lettuce

    Science.gov (United States)

    Moyne, Anne-Laure; Harris, Linda J.; Marco, Maria L.

    2013-01-01

    Leafy green produce has been associated with numerous outbreaks of foodborne illness caused by strains of Escherichia coli O157:H7. While the amounts of culturable E. coli O157:H7 rapidly decline after introduction onto lettuce in the field, it remains to be determined whether the reduction in cell numbers is due to losses in cell viability, cell injury and a subsequent inability to be detected by standard laboratory culturing methods, or a lack of adherence and hence rapid removal of the organism from the plants during application. To assess which of these options is most relevant for E. coli O157:H7 on leafy green produce, we developed and applied a propidium monoazide (PMA) real-time PCR assay to quantify viable (with PMA) and total (without PMA) E. coli O157:H7 cells on growth chamber and field-grown lettuce. E. coli O157:H7, suspended in 0.1% peptone, was inoculated onto 4-week-old lettuce plants at a level of approximately 106 CFU/plant. In the growth chamber at low relative humidity (30%), culturable amounts of the nontoxigenic E. coli O157:H7 strain ATCC 700728 and the virulent strain EC4045 declined 100 to 1000-fold in 24 h. Fewer E. coli O157:H7 cells survived when applied onto plants in droplets with a pipette compared with a fine spray inoculation. Total cells for both strains were equivalent to inoculum levels for 7 days after application, and viable cell quantities determined by PMA real-time PCR were approximately 104 greater than found by colony enumeration. Within 2 h after application onto plants in the field, the number of culturable E. coli ATCC 700728 was reduced by up to 1000-fold, whereas PCR-based assessments showed that total cell amounts were equivalent to inoculum levels. These findings show that shortly after inoculation onto plants, the majority of E. coli O157:H7 cells either die or are no longer culturable. PMID:23936235

  1. Melanocortin agonism as a viable strategy to control alveolar bone loss induced by oral infection.

    Science.gov (United States)

    Madeira, Mila F M; Queiroz-Junior, Celso M; Montero-Melendez, Trinidad; Werneck, Silvia M C; Corrêa, Jôice D; Soriani, Frederico M; Garlet, Gustavo P; Souza, Daniele G; Teixeira, Mauro M; Silva, Tarcilia A; Perretti, Mauro

    2016-12-01

    Alveolar bone loss is a result of an aggressive form of periodontal disease (PD) associated with Aggregatibacter actinomycetemcomitans (Aa) infection. PD is often observed with other systemic inflammatory conditions, including arthritis. Melanocortin peptides activate specific receptors to exert antiarthritic properties, avoiding excessing inflammation and modulating macrophage function. Recent work has indicated that melanocortin can control osteoclast development and function, but whether such protection takes place in infection-induced alveolar bone loss has not been investigated. The purpose of this study was to evaluate the role of melanocortin in Aa-induced PD. Mice were orally infected with Aa and treated with the melanocortin analog DTrp8-γMSH or vehicle daily for 30 d. Then, periodontal tissue was collected and analyzed. Aa-infected mice treated with DTrp8-γMSH presented decreased alveolar bone loss and a lower degree of neutrophil infiltration in the periodontium than vehicle-treated animals; these actions were associated with reduced periodontal levels of TNF-α, IFN-γ, and IL-17A. In vitro experiments with cells differentiated into osteoclasts showed that osteoclast formation and resorptive activity were attenuated after treatment with DTrp8-γMSH. Thus, melanocortin agonism could represent an innovative way to tame overexuberant inflammation and, at the same time, preserve bone physiology, as seen after Aa infection.-Madeira, M. F. M., Queiroz-Junior, C. M., Montero-Melendez, T., Werneck, S. M. C., Corrêa, J. D., Soriani, F. M., Garlet, G. P., Souza, D. G., Teixeira, M. M., Silva, T. A., Perretti, M. Melanocortin agonism as a viable strategy to control alveolar bone loss induced by oral infection. © FASEB.

  2. Electrochemical Stripping to Recover Nitrogen from Source-Separated Urine.

    Science.gov (United States)

    Tarpeh, William A; Barazesh, James M; Cath, Tzahi Y; Nelson, Kara L

    2018-01-19

    Recovering nitrogen from separately collected urine can potentially reduce costs and energy of wastewater nitrogen removal and fertilizer production. Through benchtop experiments, we demonstrate the recovery of nitrogen from urine as ammonium sulfate using electrochemical stripping, a combination of electrodialysis and membrane stripping. Nitrogen was selectively recovered with 93% efficiency in batch experiments with real urine and required 30.6 MJ kg N-1 in continuous-flow experiments (slightly less than conventional ammonia stripping). The effects of solution chemistry on nitrogen flux, electrolytic reactions, and reactions with electro-generated oxidants were evaluated using synthetic urine solutions. Fates of urine-relevant trace organic contaminants, including electrochemical oxidation and reaction with electro-generated chlorine, were investigated with a suite of common pharmaceuticals. Trace organics (fertilizer product. This novel approach holds promise for selective recovery of nitrogen from concentrated liquid waste streams such as source-separated urine.

  3. Recovering and recycling uranium used for production of molybdenum-99

    Energy Technology Data Exchange (ETDEWEB)

    Reilly, Sean Douglas; May, Iain; Copping, Roy; Dale, Gregory Edward

    2017-12-12

    A processes for recycling uranium that has been used for the production of molybdenum-99 involves irradiating a solution of uranium suitable for forming fission products including molybdenum-99, conditioning the irradiated solution to one suitable for inducing the formation of crystals of uranyl nitrate hydrates, then forming the crystals and a supernatant and then separating the crystals from the supernatant, thus using the crystals as a source of uranium for recycle. Molybdenum-99 is recovered from the supernatant using an adsorbent such as alumina. Another process involves irradiation of a solid target comprising uranium, forming an acidic solution from the irradiated target suitable for inducing the formation of crystals of uranyl nitrate hydrates, then forming the crystals and a supernatant and then separating the crystals from the supernatant, thus using the crystals as a source of uranium for recycle. Molybdenum-99 is recovered from the supernatant using an adsorbent such as alumina.

  4. Veterinary Forensic Pathology: Drowning and Bodies Recovered From Water.

    Science.gov (United States)

    McEwen, B J; Gerdin, J

    2016-09-01

    Determining the cause of death in animals recovered from bodies of water, swimming pools, or other water-containing vessels is challenging. Animals recovered from water may or may not have drowned. The diagnosis of drowning is usually one of exclusion, requiring information from the crime scene, recovery scene, the medical history or reliable witness accounts. While there are characteristic macroscopic and microscopic lesions of drowning, none are specific and are dependent on the volume and tonicity of the drowning medium. Beyond interpreting the postmortem findings, the court may ask pathologists to comment on the behavioral and welfare implications of drowning. This requires an understanding of the drowning process, which is a complex series of sequential, concurrent, and overlapping cardiorespiratory reflexes, electrolyte and blood gas abnormalities, aspiration, physical exhaustion, and breathlessness eventually culminating in death. This review addresses the mechanisms, lesions, and diagnostic issues associated with drowning in nonaquatic companion animals. © The Author(s) 2016.

  5. The susceptible-infected-recovered (SIR) model for viral marketing

    Science.gov (United States)

    Ismail, Siti Suhaila; Akil, Ku Azlina Ku; Chulan, Majdah; Sharif, Noorzila

    2017-11-01

    Viral marketing is a marketing strategy utilizes social media to spread information about a product or services provided. It is the most powerful way to share information in a short amount of time. The objective of this study is to investigate the dynamic of viral marketing within a time duration in the point of view of mathematics. This study used the epidemiological model known as Susceptible-Infected-Recovered (SIR). The model consists of a system of three differential equations with three state variables namely susceptible (S), infected (I) and recovered (R). It considers a case of SIR model with demography. Numerical experiments have been performed. The results show that viral marketing reaches its peak within two days. The online messages shared will become higher if the initial number of the infected individual has been increased.

  6. Serotypes of Yersinia pseudotuberculosis recovered from domestic livestock.

    Science.gov (United States)

    Hodges, R T; Carman, M G; Mortimer, W J

    1984-01-01

    The serological identity of 234 strains of Yersinia pseudotuberculosis recovered from domestic animals and birds in New Zealand was determined by slide agglutination test. Thirty strains were also examined by tube agglutination test. The strains were isolated from cattle (56), sheep (8), deer (117), goats (13), pigs (7), rabbits (6), guinea pigs (5), and aviary species of birds (22). All strains were isolated from animals or birds which had died or shown signs of ill health and amongst which diarrhoea was a common feature. Serotype I accounted for 23% (53) of strains, serotype II for 13% (30) of strains and serotype III for 64% (151) of strains. It was concluded that further investigations on the prevalence and serological identity of strains recovered from clinically healthy animals mav provide useful information in assessing the significance of various serotypes as a cause of disease in livestock.

  7. Experimental human-like model to assess the part of viable Legionella reaching the thoracic region after nebulization.

    Directory of Open Access Journals (Sweden)

    Jérémie Pourchez

    Full Text Available The incidence of Legionnaires' disease (LD in European countries and the USA has been constantly increasing since 1998. Infection of humans occurs through aerosol inhalation. To bridge the existing gap between the concentration of Legionella in a water network and the deposition of bacteria within the thoracic region (assessment of the number of viable Legionella, we validated a model mimicking realistic exposure through the use of (i recent technology for aerosol generation and (ii a 3D replicate of the human upper respiratory tract. The model's sensitivity was determined by monitoring the deposition of (i aerosolized water and Tc99m radio-aerosol as controls, and (ii bioaerosols generated from both Escherichia coli and Legionella pneumophila sg 1 suspensions. The numbers of viable Legionella prior to and after nebulization were provided by culture, flow cytometry and qPCR. This study was designed to obtain more realistic data on aerosol inhalation (vs. animal experimentation and deposition at the thoracic region in the context of LD. Upon nebulization, 40% and 48% of the initial Legionella inoculum was made of cultivable and non-cultivable cells, respectively; 0.7% of both populations reached the filter holder mimicking the thoracic region in this setup. These results are in agreement with experimental data based on quantitative microbial risk assessment methods and bring new methods that may be useful for preventing LD.

  8. PROCESS FOR RECOVERING URANIUM FROM AQUEOUS PHOSPHORIC ACID LIQUORS

    Science.gov (United States)

    Schmitt, J.M.

    1962-09-01

    A liquid-liquid extraction method is given for recovering uranium values from aqueous solutions. An acidic aqueous solution containing uranium values is contacted with an organic phase comprising an organic diluent and the reaction product of phosphorous pentoxide and a substantially pure dialkylphosphoric acid. The uranium values are transferred to the organic phase even from aqueous solutions containing a high concentration of strong uranium complexing agents such as phosphate ions. (AEC)

  9. CPR in the neonatal foal: has RECOVER changed our approach?

    Science.gov (United States)

    Jokisalo, Jonna Maaria; Corley, Kevin Thomas Trent

    2014-08-01

    RECOVER was created to optimize survival of small animal patients from cardiopulmonary arrest. Several findings from this study are applicable to cardiopulmonary resuscitation in the neonatal foal. In particular, chest compressions should be a priority with no pauses and a "push hard, push fast" approach. The importance of ventilation is minimized with short, infrequent breaths at a rate of 10 to 20 per minute recommended. Copyright © 2014 Elsevier Inc. All rights reserved.

  10. Emotional intelligence and recovering from induced negative emotional state

    OpenAIRE

    Limonero, Joaquín T.; Jordi eFernández-Castro; Jordi eSoler-Oritja; María eÁlvarez

    2015-01-01

    The aim of the present study was to examine the relationship between emotional intelligence (EI) and recovering from negative emotions induction, using a performance test to measure EI. Sixty seven undergraduates participated in the procedure, which lasted 75 min and was divided into three stages. At Time 1, subjects answered the State-Trait Anxiety Inventory (STAI)-S, Profile of Mood States (POMS)-A, and EI was assessed by Mayer–Salovey–Caruso Emotional Intelligence Test (MSCEIT). At Time 2,...

  11. Methods for recovering precious metals from industrial waste

    Science.gov (United States)

    Canda, L.; Heput, T.; Ardelean, E.

    2016-02-01

    The accelerated rate of industrialization increases the demand for precious metals, while high quality natural resources are diminished quantitatively, with significant operating costs. Precious metals recovery can be successfully made from waste, considered to be secondary sources of raw material. In recent years, concerns and interest of researchers for more increasing efficient methods to recover these metals, taking into account the more severe environmental protection legislation. Precious metals are used in a wide range of applications, both in electronic and communications equipment, spacecraft and jet aircraft engines and for mobile phones or catalytic converters. The most commonly recovered precious metals are: gold from jewellery and electronics, silver from X- ray films and photographic emulsions, industrial applications (catalysts, batteries, glass/mirrors), jewellery; platinum group metals from catalytic converters, catalysts for the refining of crude oil, industrial catalysts, nitric acid manufacturing plant, the carbon-based catalyst, e-waste. An important aspect is the economic viability of recycling processes related to complex waste flows. Hydrometallurgical and pyrometallurgical routes are the most important ways of processing electrical and electronic equipment waste. The necessity of recovering precious metals has opened new opportunities for future research.

  12. Flexible variable selection for recovering sparsity in nonadditive nonparametric models.

    Science.gov (United States)

    Fang, Zaili; Kim, Inyoung; Schaumont, Patrick

    2016-12-01

    Variable selection for recovering sparsity in nonadditive and nonparametric models with high-dimensional variables has been challenging. This problem becomes even more difficult due to complications in modeling unknown interaction terms among high-dimensional variables. There is currently no variable selection method to overcome these limitations. Hence, in this article we propose a variable selection approach that is developed by connecting a kernel machine with the nonparametric regression model. The advantages of our approach are that it can: (i) recover the sparsity; (ii) automatically model unknown and complicated interactions; (iii) connect with several existing approaches including linear nonnegative garrote and multiple kernel learning; and (iv) provide flexibility for both additive and nonadditive nonparametric models. Our approach can be viewed as a nonlinear version of a nonnegative garrote method. We model the smoothing function by a Least Squares Kernel Machine (LSKM) and construct the nonnegative garrote objective function as the function of the sparse scale parameters of kernel machine to recover sparsity of input variables whose relevances to the response are measured by the scale parameters. We also provide the asymptotic properties of our approach. We show that sparsistency is satisfied with consistent initial kernel function coefficients under certain conditions. An efficient coordinate descent/backfitting algorithm is developed. A resampling procedure for our variable selection methodology is also proposed to improve the power. © 2016, The International Biometric Society.

  13. Diffraction study of pressure-amorphized ZrW2O8 using in situ and recovered samples

    Science.gov (United States)

    Keen, David A.; Goodwin, Andrew L.; Tucker, Matthew G.; Hriljac, Joseph A.; Bennett, Thomas D.; Dove, Martin T.; Kleppe, Annette K.; Jephcoat, Andrew P.; Brunelli, Michela

    2011-02-01

    High-energy x-ray diffraction data from a series of amorphous ZrW2O8 samples, recovered from a number of different pressures, have been measured and compared with similar data collected from a sample contained within a diamond anvil cell and measured in situ at high pressure. Samples at lower pressures are a mix of crystalline and amorphous phases, whereas those above 7 GPa (both recovered and in situ) appear to be completely amorphous. Although there are only very small changes in the diffraction data from the pure phase samples recovered from different pressures, larger changes are observed in situ as a function of pressure. However, the same structural model can be used to describe all data, albeit using different densities. This strongly suggests that the amorphous phase is undergoing nonreconstructive compression in this pressure range, without fundamental modification of the underlying topology.

  14. On the use of the serial dilution culture method to enumerate viable phytoplankton in natural communities of plankton subjected to ballast water treatment

    OpenAIRE

    Cullen, John J.; MacIntyre, Hugh L.

    2015-01-01

    Discharge standards for ballast water treatment (BWT) systems are based on concentrations of living cells, for example, as determined with vital stains. Ultraviolet radiation (UV) stops the reproduction of microorganisms without killing them outright; they are living, but not viable, and ecologically as good as dead. Consequently, UV-treated discharge can be compliant with the intent of regulation while failing a live/dead test. An alternative evaluation of BWT can be proposed based on the as...

  15. Allogeneic cardiospheres delivered via percutaneous transendocardial injection increase viable myocardium, decrease scar size, and attenuate cardiac dilatation in porcine ischemic cardiomyopathy.

    Directory of Open Access Journals (Sweden)

    Kristine Yee

    Full Text Available Epicardial injection of heart-derived cell products is safe and effective post-myocardial infarction (MI, but clinically-translatable transendocardial injection has never been evaluated. We sought to assess the feasibility, safety and efficacy of percutaneous transendocardial injection of heart-derived cells in porcine chronic ischemic cardiomyopathy.We studied a total of 89 minipigs; 63 completed the specified protocols. After NOGA-guided transendocardial injection, we quantified engraftment of escalating doses of allogeneic cardiospheres or cardiosphere-derived cells in minipigs (n = 22 post-MI. Next, a dose-ranging, blinded, randomized, placebo-controlled ("dose optimization" study of transendocardial injection of the better-engrafting product was performed in infarcted minipigs (n = 16. Finally, the superior product and dose (150 million cardiospheres were tested in a blinded, randomized, placebo-controlled ("pivotal" study (n = 22. Contrast-enhanced cardiac MRI revealed that all cardiosphere doses preserved systolic function and attenuated remodeling. The maximum feasible dose (150 million cells was most effective in reducing scar size, increasing viable myocardium and improving ejection fraction. In the pivotal study, eight weeks post-injection, histopathology demonstrated no excess inflammation, and no myocyte hypertrophy, in treated minipigs versus controls. No alloreactive donor-specific antibodies developed over time. MRI showed reduced scar size, increased viable mass, and attenuation of cardiac dilatation with no effect on ejection fraction in the treated group compared to placebo.Dose-optimized injection of allogeneic cardiospheres is safe, decreases scar size, increases viable myocardium, and attenuates cardiac dilatation in porcine chronic ischemic cardiomyopathy. The decreases in scar size, mirrored by increases in viable myocardium, are consistent with therapeutic regeneration.

  16. Immunization of Rodents Against Hymenolepis Infections using Non-Viable Homologous Oncospheres

    Directory of Open Access Journals (Sweden)

    Ping-Chin Fan

    2004-12-01

    Full Text Available Immunity to Taiwan Taenia infection in pigs can be stimulated using homologous or heterologous nonviable Taenia oncospheres. This study was designed to determine whether homologous non-viable oncospheres could stimulate immunity to Hymenolepis infection in rodents. Hatched oncospheres were prepared from eggs of Hymenolepis diminuta, Hymenolepis nana, and Hymenolepis microstoma and kept at −70°C for more than 1 month. A mixture of 500 non-viable oncospheres of each tapeworm and complete Freund's adjuvant was injected subcutaneously in four groups of Sprague-Dawley rats or ICR mice one to four times at an interval of 1 week; controls were not immunized. After immunization, each rodent was orally inoculated with three fresh active cysticercoids of H. diminuta or H. microstoma or 500 fresh eggs of H. nana. The animals were then necropsied for adult tapeworms. No rats or mice immunized with non-viable oncospheres of H. diminuta or H. nana were infected by the challenge inoculation. However, 28 of 34 mice immunized with non-viable H. microstoma oncospheres were infected after inoculation with cysticercoids. This study demonstrated complete protection against infection by homologous parasites in rats or mice immunized with non-viable oncospheres of H. diminuta and H. nana, respectively. Repeated immunization may not be required if resistance is stimulated in rodent hosts.

  17. Airborne viable fungi in school environments in different climatic regions - A review

    Science.gov (United States)

    Salonen, Heidi; Duchaine, Caroline; Mazaheri, Mandana; Clifford, Sam; Lappalainen, Sanna; Reijula, Kari; Morawska, Lidia

    2015-03-01

    Elevated levels of fungi in indoor environments have been linked with mould/moisture damage in building structures. However, there is a lack of information about "normal" concentrations and flora as well as guidelines of viable fungi in the school environment in different climatic conditions. We have reviewed existing guidelines for indoor fungi and the current knowledge of the concentrations and flora of viable fungi in different climatic areas, the impact of the local factors on concentrations and flora of viable fungi in school environments. Meta-regression was performed to estimate the average behaviour for each analysis of interest, showing wide variation in the mean concentrations in outdoor and indoor school environments (range: 101-103 cfu/m3). These concentrations were significantly higher for both outdoors and indoors in the moderate than in the continental climatic area, showing that the climatic condition was a determinant for the concentrations of airborne viable fungi. The most common fungal species both in the moderate and continental area were Cladosporium spp. and Penicillium spp. The suggested few quantitative guidelines for indoor air viable fungi for school buildings are much lower than for residential areas. This review provides a synthesis, which can be used to guide the interpretation of the fungi measurements results and help to find indications of mould/moisture in school building structures.

  18. First study on the formation and resuscitation of viable but nonculturable state and beer spoilage capability of Lactobacillus lindneri.

    Science.gov (United States)

    Liu, Junyan; Li, Lin; Li, Bing; Peters, Brian M; Deng, Yang; Xu, Zhenbo; Shirtliff, Mark E

    2017-06-01

    This study aimed to investigate the spoilage capability of Lactobacillus lindneri during the induction and resuscitation of viable but nonculturable (VBNC) state. L. lindneri strain was identified by sequencing the PCR product (amplifying 16S rRNA gene) using ABI Prism 377 DNA Sequencer. During the VBNC state induction by low temperature storage and beer adaption, total, culturable, and viable cells were assessed by acridine orange direct counting, plate counting, and Live/Dead BacLight bacterial viability kit, respectively. Organic acids and diacetyl concentration were measured by reversed-phase high performance liquid chromatography and head dpace gas chromatography, respectively. VBNC state of L. lindneri was successfully induced by both beer adaption and low temperature storage, and glycerol frozen stock was the optimal way to maintain the VBNC state. Addition of catalase was found to be an effective method for the resuscitation of VBNC L. lindneri cells. Furthermore, spoilage capability remained similar during the induction and resuscitation of VBNC L. lindneri. This is the first report of induction by low temperature storage and resuscitation of VBNC L. lindneri strain, as well as the first identification of spoilage capability of VBNC and resuscitated L. lindneri cells. This study indicated that the potential colonization of L. lindneri strain in brewery environment, formation and resuscitation of VBNC state, as well as maintenance in beer spoilage capability, may be an important risk factor for brewery environment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. A Transient Cell-shielding Method for Viable MSC Delivery Within Hydrophobic Scaffolds Polymerized in situ

    Science.gov (United States)

    2015-03-27

    reporting burden for the collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching...existing data sources, gathering and maintaining the data needed, and completing and reviewing the collection of information. Send comments regarding this...Celzard A. Effect of composition and pro- cessing parameters on the characteristics of tannin -based rigid foams. Part II: physical properties. Mater

  20. A Novel Method for Determining the Level of Viable Disseminated Prostate Cancer Cells

    Science.gov (United States)

    2012-10-01

    Finally, in this final vector, we have introduced an integrin-targeting peptide ( RGD ) into the Fiber protein to broaden the tropism of the virus...3077. 22. Lupold SE, Kudrolli TA, Chowdhury WH, Wu P, Rodriguez R (2007) A novel method for generating and screening peptides and libraries displayed

  1. Cryopreserved ovarian cortex from patients with leukemia in complete remission contains no apparent viable malignant cells

    DEFF Research Database (Denmark)

    Greve, Tine; Clasen-Linde, Erik; Andersen, Morten T

    2012-01-01

    Some women suffering from leukemia require bone marrow transplantation to be cured. Bone marrow transplantation is associated with a high risk of sterility and some patients are offered fertility preservation by cryopreservation of ovarian cortex. Transplantation of ovarian cortex to women cured...

  2. A multicenter study of viable PCR using propidium monoazide to detect Legionella in water samples.

    Science.gov (United States)

    Scaturro, Maria; Fontana, Stefano; Dell'eva, Italo; Helfer, Fabrizia; Marchio, Michele; Stefanetti, Maria Vittoria; Cavallaro, Mario; Miglietta, Marilena; Montagna, Maria Teresa; De Giglio, Osvalda; Cuna, Teresa; Chetti, Leonarda; Sabattini, Maria Antonietta Bucci; Carlotti, Michela; Viggiani, Mariagabriella; Stenico, Alberta; Romanin, Elisa; Bonanni, Emma; Ottaviano, Claudio; Franzin, Laura; Avanzini, Claudio; Demarie, Valerio; Corbella, Marta; Cambieri, Patrizia; Marone, Piero; Rota, Maria Cristina; Bella, Antonino; Ricci, Maria Luisa

    2016-07-01

    Legionella quantification in environmental samples is overestimated by qPCR. Combination with a viable dye, such as Propidium monoazide (PMA), could make qPCR (named then vPCR) very reliable. In this multicentre study 717 artificial water samples, spiked with fixed concentrations of Legionella and interfering bacterial flora, were analysed by qPCR, vPCR and culture and data were compared by statistical analysis. A heat-treatment at 55 °C for 10 minutes was also performed to obtain viable and not-viable bacteria. When data of vPCR were compared with those of culture and qPCR, statistical analysis showed significant differences (P 0.05). Overall this study provided a good experimental reproducibility of vPCR but also highlighted limits of PMA in the discriminating capability of dead and live bacteria, making vPCR not completely reliable. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. Issues of organizational cybernetics and viability beyond Beer's viable systems model

    Science.gov (United States)

    Nechansky, Helmut

    2013-11-01

    The paper starts summarizing the claims of Beer's viable systems model to identify five issues any viable organizations has to deal with in an unequivocal hierarchical structure of five interrelated systems. Then the evidence is introduced for additional issues and related viable structures of organizations, which deviate from Beer's model. These issues are: (1) the establishment and (2) evolution of an organization; (3) systems for independent top-down control (like "Six Sigma"); (4) systems for independent bottom-up correction of performance problems (like "Kaizen"), both working outside a hierarchical structure; (5) pull production systems ("Just in Time") and (6) systems for checks and balances of top-level power (like boards and shareholder meetings). Based on that an evolutionary approach to organizational cybernetics is outlined, addressing the establishment of organizations and possible courses of developments, including recent developments in quality and production engineering, as well as problems of setting and changing goal values determining organizational policies.

  4. El modelo de sistema viable: un instrumento para la organización efectiva

    Directory of Open Access Journals (Sweden)

    Norlando Sánchez Rueda

    2015-05-01

    Full Text Available RESUMEN En este ensayo se presenta una interpretación teórica del denominado Modelo de Sistema Viable (MSV, de Stafford Beer y su Potencial Aplicación en Tareas de Diagnóstico  y diseño empresarial, al igual que para Mejorar las capacidades Organizacionales de Auto- Regulación  y Auto- Organización. Se explica como el Modelo del Sistema Viable permite conocer e interpretar  los mecanismos de estabilidad y adaptabilidad de las organizaciones, pilares para el crecimiento de una verdadera organización Efectiva.

  5. Fractionation of transgenic corn seed by dry and wet milling to recover recombinant collagen-related proteins.

    Science.gov (United States)

    Zhang, Cheng; Glatz, Charles E; Fox, Steven R; Johnson, Lawrence A

    2009-01-01

    Corn continues to be considered an attractive transgenic host for producing recombinant therapeutic and industrial proteins because of its potential for producing recombinant proteins at large volume and low cost as coproducts of corn seed-based biorefining. Efforts to reduce production costs have been primarily devoted to increasing accumulation level, optimizing protein extraction conditions, and simplifying the purification. In the present work, we evaluated two grain fractionation methods, dry milling and wet milling, to enrich two recombinant collagen-related proteins; thereby, reducing the amount and type of corn-derived impurities in subsequent protein extraction and purification steps. The two proteins were a full-length human recombinant collagen type I alpha 1(rCIalpha1) chain with telopeptides and peptide foldon to effect triple helix formation and a 44-kDa rCIalpha1 fragment. For each, approximately 60% of the rCIalpha1s in the seed was recovered in the dry-milled germ-rich fractions making up ca. 25% of the total kernel mass. For wet milling, approximately 60% of each was recovered in three fractions accounting for 20-25% of the total kernel mass. The rCIalpha1s in the dry-milled germ-rich fractions were enriched three to six times compared with the whole corn kernel, whereas the rCIalpha1s were enriched 4-10 times in selected wet-milled fractions. The recovered starch from wet milling was almost free of rCIalpha1. Therefore, it was possible to generate rCIalpha1-enriched fractions by both dry and wet milling along with rCIalpha1-free starch using wet milling. Because of its simplicity, the dry milling procedure could be accomplished on-farm thus minimizing the risk of inadvertent release of viable transgenic seeds. 2009 American Institute of Chemical Engineers Biotechnol.

  6. Optimization of Culture Medium Enhances Viable Biomass Production and Biocontrol Efficacy of the Antagonistic Yeast, Candida diversa

    Directory of Open Access Journals (Sweden)

    Jia Liu

    2017-10-01

    Full Text Available Viable biomass production is a key determinant of suitability of antagonistic yeasts as potential biocontrol agents. This study investigated the effects of three metal ions (magnesium, ferrous, and zinc on biomass production and viability of the antagonistic yeast, Candida diversa. Using response surface methodology to optimize medium components, a maximum biomass was obtained, when the collective Mg2+, Fe2+, and Zn2+ concentrations were adjusted in a minimal mineral (MM medium. Compared with the unmodified MM, and three ion-deficient MM media, yeast cells cultured in the three ion-modified MM medium exhibited a lower level of cellular oxidative damage, and a higher level of antioxidant enzyme activity. A biocontrol assay indicated that C. diversa grown in the ion-modified MM exhibited the greatest level of control of gray mold on apple fruit. These results provide new information on culture medium optimization to grow yeast antagonists in order to improve biomass production and biocontrol efficacy.

  7. Optimization of Culture Medium Enhances Viable Biomass Production and Biocontrol Efficacy of the Antagonistic Yeast, Candida diversa.

    Science.gov (United States)

    Liu, Jia; Li, Guangkun; Sui, Yuan

    2017-01-01

    Viable biomass production is a key determinant of suitability of antagonistic yeasts as potential biocontrol agents. This study investigated the effects of three metal ions (magnesium, ferrous, and zinc) on biomass production and viability of the antagonistic yeast, Candida diversa. Using response surface methodology to optimize medium components, a maximum biomass was obtained, when the collective Mg(2+), Fe(2+), and Zn(2+) concentrations were adjusted in a minimal mineral (MM) medium. Compared with the unmodified MM, and three ion-deficient MM media, yeast cells cultured in the three ion-modified MM medium exhibited a lower level of cellular oxidative damage, and a higher level of antioxidant enzyme activity. A biocontrol assay indicated that C. diversa grown in the ion-modified MM exhibited the greatest level of control of gray mold on apple fruit. These results provide new information on culture medium optimization to grow yeast antagonists in order to improve biomass production and biocontrol efficacy.

  8. On the use of the serial dilution culture method to enumerate viable phytoplankton in natural communities of plankton subjected to ballast water treatment.

    Science.gov (United States)

    Cullen, John J; MacIntyre, Hugh L

    2016-01-01

    Discharge standards for ballast water treatment (BWT) systems are based on concentrations of living cells, for example, as determined with vital stains. Ultraviolet radiation (UV) stops the reproduction of microorganisms without killing them outright; they are living, but not viable, and ecologically as good as dead. Consequently, UV-treated discharge can be compliant with the intent of regulation while failing a live/dead test. An alternative evaluation of BWT can be proposed based on the assessment of viable, rather than living, cells in discharge water. In principle, the serial dilution culture-most probable number (SDC-MPN) method provides the appropriate measure for phytoplankton. But, the method has been criticized, particularly because it is thought that many phytoplankton species cannot be cultured. A review of the literature shows that although SDC-MPN has been used for more than 50 years-generally to identify and count phytoplankton species that cannot be preserved-its application to enumerate total viable phytoplankton seems to be new, putting past criticisms of the method in a different light. Importantly, viable cells need to grow only enough to be detected, not to be brought into sustained culture, and competition between species in a dilution tube is irrelevant as long as the winner is detectable. Thorough consideration of sources of error leads to recommendations for minimizing and quantifying uncertainties by optimizing growth conditions and conducting systematic comparisons. We conclude that with careful evaluation, SDC-MPN is potentially an effective method for assessing the viability of phytoplankton after BWT.

  9. Rapid quantification of viable Campylobacter bacteria on chicken carcasses, using real-time PCR and propidium monoazide treatment, as a tool for quantitative risk assessment.

    Science.gov (United States)

    Josefsen, M H; Löfström, C; Hansen, T B; Christensen, L S; Olsen, J E; Hoorfar, J

    2010-08-01

    A number of intervention strategies against Campylobacter-contaminated poultry focus on postslaughter reduction of the number of cells, emphasizing the need for rapid and reliable quantitative detection of only viable Campylobacter bacteria. We present a new and rapid quantitative approach to the enumeration of food-borne Campylobacter bacteria that combines real-time quantitative PCR (Q-PCR) with simple propidium monoazide (PMA) sample treatment. In less than 3 h, this method generates a signal from only viable and viable but nonculturable (VBNC) Campylobacter bacteria with an intact membrane. The method's performance was evaluated by assessing the contributions to variability by individual chicken carcass rinse matrices, species of Campylobacter, and differences in efficiency of DNA extraction with differing cell inputs. The method was compared with culture-based enumeration on 50 naturally infected chickens. The cell contents correlated with cycle threshold (C(T)) values (R(2) = 0.993), with a quantification range of 1 x 10(2) to 1 x 10(7) CFU/ml. The correlation between the Campylobacter counts obtained by PMA-PCR and culture on naturally contaminated chickens was high (R(2) = 0.844). The amplification efficiency of the Q-PCR method was not affected by the chicken rinse matrix or by the species of Campylobacter. No Q-PCR signals were obtained from artificially inoculated chicken rinse when PMA sample treatment was applied. In conclusion, this study presents a rapid tool for producing reliable quantitative data on viable Campylobacter bacteria in chicken carcass rinse. The proposed method does not detect DNA from dead Campylobacter bacteria but recognizes the infectious potential of the VBNC state and is thereby able to assess the effect of control strategies and provide trustworthy data for risk assessment.

  10. Gainful employment reduces stigma toward people recovering from schizophrenia.

    Science.gov (United States)

    Perkins, David V; Raines, Joshua A; Tschopp, Molly K; Warner, Todd C

    2009-06-01

    Stigma impedes the social integration of persons recovering from psychiatric disability, especially those with criminal histories. Little is known about factors that lessen this stigma. Four hundred and four adults listened to one of four vignettes describing a 25-year-old male with schizophrenia and responded to a standard set of items measuring social distance. The individual who was gainfully employed (vs. unemployed), or who had a prior misdemeanor (vs. felony) criminal offense, elicited significantly less stigma. Employment may destigmatize a person coping with both psychiatric disability and a criminal record. Mental health services should encourage paid employment and other paths to community integration.

  11. Comparison of sampling methods to recover germinated Bacillus anthracis and Bacillus thuringiensis endospores from surface coupons.

    Science.gov (United States)

    Mott, T M; Shoe, J L; Hunter, M; Woodson, A M; Fritts, K A; Klimko, C P; Quirk, A V; Welkos, S L; Cote, C K

    2017-05-01

    In an attempt to devise decontamination methods that are both effective and minimally detrimental to the environment, we evaluated germination induction as an enhancement to strategies for Bacillus anthracis spore decontamination. To determine an optimal method for the recovery of germinating spores from different matrices, it was critical to ensure that the sampling procedures did not negatively impact the viability of the germinating spores possibly confounding the results and downstream analyses of field trial data. Therefore, the two main objectives of this study were the following: (i) development of an effective processing protocol capable of recovering the maximum number of viable germinating or germinated spores from different surface materials; and (ii) using a model system of spore contamination, employ this protocol to evaluate the potential applicability of germination induction to wide-area decontamination of B. anthracis spores. We examined parameters affecting the sampling efficiencies of B. anthracis and the surrogate species Bacillus thuringiensis on nonporous and porous materials. The most efficient extraction from all matrices was observed using PBS with 0·01% Tween 80 extraction buffer. The addition of a sonication and/or extended vortex treatment did not yield significant increases in spore or germinated spore recovery. Our data demonstrate that previous germination-induction experiments performed in suspension can be reproduced when Bacillus spores are deposited onto reference surfaces materials. Our proof of concept experiment illustrated that a germination pretreatment step significantly improves conventional secondary decontamination strategies and remediation plans. Published 2017. This article is a U.S. Government work and is in the public domain in the USA.

  12. Induction of viable 2n pollen in sterile Oriental × Trumpet Lilium hybrids

    NARCIS (Netherlands)

    Luo, J.R.; Arens, P.; Niu, L.X.; Tuyl, van J.M.

    2016-01-01

    In order to induce viable 2n pollen from highly sterile diploid Oriental × Trumpet (OT) (Lilium), N2O was used to treat flower buds of four sterile diploid OT cultivars (‘Nymph’, ‘Gluhwein’, ‘Yelloween’, and ‘Shocking’) at different stages of meiosis. There was no pollen germination in

  13. 9 CFR 113.26 - Detection of viable bacteria and fungi except in live vaccine.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Detection of viable bacteria and fungi except in live vaccine. 113.26 Section 113.26 Animals and Animal Products ANIMAL AND PLANT HEALTH... in live vaccine. Each serial and subserial of biological product except live vaccines shall be tested...

  14. Oligonucleotide microarrays for the detection and identification of viable beer spoilage bacteria.

    Science.gov (United States)

    Weber, D G; Sahm, K; Polen, T; Wendisch, V F; Antranikian, G

    2008-10-01

    The design and evaluation of an oligonucleotide microarray in order to detect and identify viable bacterial species that play a significant role in beer spoilage. These belong to the species of the genera Lactobacillus, Megasphaera, Pediococcus and Pectinatus. Oligonucleotide probes specific to beer spoilage bacteria were designed. In order to detect viable bacteria, the probes were designed to target the intergenic spacer regions (ISR) between 16S and 23S rRNA. Prior to hybridization the ISR were amplified by combining reverse transcriptase and polymerase chain reactions using a designed consenus primer. The developed oligonucleotide microarrays allows the detection of viable beer spoilage bacteria. This method allows the detection and discrimination of single bacterial species in a sample containing complex microbial community. Furthermore, microarrays using oligonucleotide probes targeting the ISR allow the distinction between viable bacteria with the potential to grow and non growing bacteria. The results demonstrate the feasibility of oligonucleotide microarrays as a contamination control in food industry for the detection and identification of spoilage micro-organisms within a mixed population.

  15. Viable Techniques, Leontief’s Closed Model, and Sraffa’s Subsistence Economies

    Directory of Open Access Journals (Sweden)

    Alberto Benítez

    2014-11-01

    Full Text Available This paper studies the production techniques employed in economies that reproduce themselves. Special attention is paid to the distinction usually made between those that do not produce a surplus and those that do, which are referred to as first and second class economies, respectively. Based on this, we present a new definition of viable economies and show that every viable economy of the second class can be represented as a viable economy of the first class under two different forms, Leontief‘s closed model and Sraffa’s subsistence economies. This allows us to present some remarks concerning the economic interpretation of the two models. On the one hand, we argue that the participation of each good in the production of every good can be considered as a normal characteristic of the first model and, on the other hand, we provide a justification for the same condition to be considered a characteristic of the second model. Furthermore, we discuss three definitions of viable techniques advanced by other authors and show that they differ from ours because they admit economies that do not reproduce themselves completely.

  16. 9 CFR 113.27 - Detection of extraneous viable bacteria and fungi in live vaccines.

    Science.gov (United States)

    2010-01-01

    ... bacteria and fungi in live vaccines. 113.27 Section 113.27 Animals and Animal Products ANIMAL AND PLANT... bacteria and fungi in live vaccines. Unless otherwise specified by the Administrator or elsewhere exempted... Seed Bacteria shall be tested for extraneous viable bacteria and fungi as prescribed in this section. A...

  17. Food web (bio-)manipulation of South African reservoirs – viable ...

    African Journals Online (AJOL)

    2006-10-04

    Oct 4, 2006 ... Food web (bio-)manipulation of South African reservoirs – viable eutrophication ... Microcystis in local eutrophic waters is perceived as a primary major constraint in implementing 'classical' food-web manip- ulation. Intrinsic ... cially in 'shallow' lakes) to dismal failure (often in deep lakes) and a range of ...

  18. Estimating when the Antarctic Ozone Hole will Recover

    Science.gov (United States)

    Newman, Paul A.; Nash, Eric R.; Douglass, Anne R.; Nielsen, J. Eric; Pawson, Steven; Stolarski, Richard S.

    2007-01-01

    The Antarctic ozone hole develops each year and culminates by early spring (late September - early October). The severity of the hole has been assessed from satellites using the minimum total ozone value from the October monthly mean (depth of the hole) and by calculating the average area coverage during this September-October period. Profile information shows that ozone is completely destroyed in the 14-2 1 km layer by early October. Ozone is mainly destroyed by halogen (chlorine and bromine) catalytic cycles, and these losses are modulated by temperature variations. Because atmospheric halogen levels are responding to international a'greements that limit or phase out production, the amount of halogens in the stratosphere should decrease over the next few decades. Using projections of halogen levels combined with age-of-air estimates, we find that the ozone hole is recovering at an extremely slow rate and that large ozone holes will regularly recur over the next 2 decades. We estimate that the ozone hole will begin to show first signs of size decrease in about 2023, and the hole will fully recover to pre-1980 levels in approximately 2070. Estimates of the ozone hole's recovery from models reveal important differences that will be discussed.

  19. Spatiotemporal inpainting for recovering texture maps of occluded building facades.

    Science.gov (United States)

    Korah, Thommen; Rasmussen, Christopher

    2007-09-01

    We present a technique for constructing a "clean" texture map of a partially occluded building facade from a series of images taken from a moving camera. Building regions blocked by trees, signs, people, and other foreground objects in a minority of views can be recovered via temporal median filtering on a registered image mosaic of the planar facade. However, when such areas are occluded in the majority of camera views, appearance information from other visible portions of the facade provides a critical cue to correctly complete the mosaic. In this paper, we apply a robust measure of spread to infer whether a particular mosaic pixel is occluded in a majority of views, and introduce a novel spatiotemporal timeline-based inpainting algorithm that uses appearance and motion cues in order to fill the texture map in majority-occluded regions. We describe methods for automatically training appearance-based classifiers from a coarse motion-based segmentation to efficiently recognize foreground and background patches in static imagery. Results of recovered building facades are shown for various sequences.

  20. How Close We Are to Achieving Commercially Viable Large-Scale Photobiological Hydrogen Production by Cyanobacteria: A Review of the Biological Aspects

    Science.gov (United States)

    Sakurai, Hidehiro; Masukawa, Hajime; Kitashima, Masaharu; Inoue, Kazuhito

    2015-01-01

    Photobiological production of H2 by cyanobacteria is considered to be an ideal source of renewable energy because the inputs, water and sunlight, are abundant. The products of photobiological systems are H2 and O2; the H2 can be used as the energy source of fuel cells, etc., which generate electricity at high efficiencies and minimal pollution, as the waste product is H2O. Overall, production of commercially viable algal fuels in any form, including biomass and biodiesel, is challenging, and the very few systems that are operational have yet to be evaluated. In this paper we will: briefly review some of the necessary conditions for economical production, summarize the reports of photobiological H2 production by cyanobacteria, present our schemes for future production, and discuss the necessity for further progress in the research needed to achieve commercially viable large-scale H2 production. PMID:25793279

  1. Chimeric rhinoviruses obtained via genetic engineering or artificially induced recombination are viable only if the polyprotein coding sequence derives from the same species.

    Science.gov (United States)

    Schibler, Manuel; Piuz, Isabelle; Hao, Weidong; Tapparel, Caroline

    2015-04-01

    Recombination is a widespread phenomenon that ensures both the stability and variation of RNA viruses. This phenomenon occurs with different frequencies within species of the Enterovirus genus. Intraspecies recombination is described frequently among non-rhinovirus enteroviruses but appears to be sporadic in rhinoviruses. Interspecies recombination is even rarer for rhinoviruses and mostly is related to ancient events which contributed to the speciation of these viruses. We reported that artificially engineered 5' untranslated region (UTR) interspecies rhinovirus/rhinovirus or rhinovirus/non-rhinovirus enterovirus recombinants are fully viable. Using a similar approach, we demonstrated in this study that exchanges of the P1-2A polyprotein region between members of the same rhinovirus species, but not between members of different species, give rise to competent chimeras. To further assess the rhinovirus intra- and interspecies recombination potential, we used artificially induced recombination by cotransfection of 5'-end-deleted and 3'-end-deleted and replication-deficient genomes. In this system, intraspecies recombination also resulted in viable viruses with high frequency, whereas no interspecies rhinovirus recombinants could be recovered. Mapping intraspecies recombination sites within the polyprotein highlighted recombinant hotspots in nonstructural genes and at gene boundaries. Notably, all recombinants occurring at gene junctions presented in-frame sequence duplications, whereas most intragenic recombinants were homologous. Taken together, our results suggest that only intraspecies recombination gives rise to viable rhinovirus chimeras in the polyprotein coding region and that recombination hotspots map to nonstructural genes with in-frame duplications at gene boundaries. These data provide new insights regarding the mechanism and limitations of rhinovirus recombination. Recombination represents a means to ensure both the stability and the variation of RNA

  2. Isolation of viable Toxoplasma gondii, molecular characterization, and seroprevalence in elk (Cervus canadensis) in Pennsylvania, USA.

    Science.gov (United States)

    Dubey, J P; Brown, J; Verma, S K; Cerqueira-Cézar, C K; Banfield, J; Kwok, O C H; Ying, Y; Murata, F H A; Pradhan, A K; Su, C

    2017-08-30

    Toxoplasmosis is a worldwide zoonosis. The ingestion of uncooked/undercooked meat and consumption of water contaminated with Toxoplasma gondii oocysts excreted by felids are the main modes of transmission of this parasite. T. gondii has been reported in multiple cervid species; however, little is known of the parasite in North American elk (Cervus canadensis). In the present study, antibodies to T. gondii were detected in serum of wild elk from Pennsylvania collected during 2013-2016 by the modified agglutination test (MAT, cut-off 1:25); 221 of 317 (69.7%) had MAT titers of 1:25 in 19, 1:50 in 28, 1:100 in 34, and 1:200 or higher in 140. Thus most (44.1%) elk had relatively high titers. Seroprevalence was slightly higher in males (76.9%) than females (67.5%, not statistically significant, Chi-square tests, P<0.0001) and was higher in adults (76.5%) than yearlings (46.4%, Odds ratio 3.82; 95% CL 1.72-8.47; P=0.001) or calves (21.7%, Odds ratio 12.58; 95% CL 4.51-35.10; P<0.0001). Annual seroprevalence was relatively stable throughout the period tested and ranged from 66.6% to 72.2%. Of the 101 elk harvested in 2016, hearts were bioassayed from 20 elk and tongues were bioassayed from 56; all tongue samples were negative. Viable T. gondii was isolated from hearts of two female elk, one of these was a seronegative adult and the other was a calf with no serum available for testing. Both T. gondii isolates were cultivated in cell culture and DNA derived from tachyzoites was characterized using the PCR-RFLP markers including SAG1, SAG2 (5'- 3'SAG2 and altSAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. One isolate belongs to ToxoDB PCR-RFLP genotype #2 and the other is genotype #5. Both genotypes are frequently identified in animals in North America. Copyright © 2017. Published by Elsevier B.V.

  3. Microfluidic Isolation of Circulating Tumor Cell Clusters by Size and Asymmetry.

    Science.gov (United States)

    Au, Sam H; Edd, Jon; Stoddard, Amy E; Wong, Keith H K; Fachin, Fabio; Maheswaran, Shyamala; Haber, Daniel A; Stott, Shannon L; Kapur, Ravi; Toner, Mehmet

    2017-05-26

    Circulating tumor cell clusters (CTC clusters) are potent initiators of metastasis and potentially useful clinical markers for patients with cancer. Although there are numerous devices developed to isolate individual circulating tumor cells from blood, these devices are ineffective at capturing CTC clusters, incapable of separating clusters from single cells and/or cause cluster damage or dissociation during processing. The only device currently able to specifically isolate CTC clusters from single CTCs and blood cells relies on the batch immobilization of clusters onto micropillars which necessitates long residence times and causes damage to clusters during release. Here, we present a two-stage continuous microfluidic chip that isolates and recovers viable CTC clusters from blood. This approach uses deterministic lateral displacement to sort clusters by capitalizing on two geometric properties: size and asymmetry. Cultured breast cancer CTC clusters containing between 2-100 + cells were recovered from whole blood using this integrated two-stage device with minimal cluster dissociation, 99% recovery of large clusters, cell viabilities over 87% and greater than five-log depletion of red blood cells. This continuous-flow cluster chip will enable further studies examining CTC clusters in research and clinical applications.

  4. Development of a specific immunomagnetic capture-PCR for rapid detection of viable Mycoplasma agalactiae in sheep milk samples.

    Science.gov (United States)

    Sanna, G; Lecca, V; Foddai, A; Tola, S

    2014-12-01

    To develop an immunomagnetic capture (IMC) to detect viable Mycoplasma agalactiae in routine ovine milk samples. Polyclonal antibodies against two M. agalactiae membrane surface proteins (P80 and P55) were covalently conjugated to magnetic beads (MBs) to form MB-Ab80 and MB-Ab55. Mycoplasma agalactiae cells were captured by a specific antigen-antibody reaction and magnetic separation. Immunomagnetic capture (IMC) was used to isolate and concentrate M. agalactiae in serial decimal dilutions and in artificially contaminated milk to facilitate subsequent detection by PCR. A 375-bp fragment of M. agalactiae was amplified using a pair of M. agalactiae-specific primers in PCR. The limit of detection of IMC-PCR method ranged from 10 to 10(2)  CCU ml(-1) when mycoplasmas were resuspended in PBS and from 10(2) to 10(3)  CCU ml(-1) when mycoplasmas were resuspended in uncontaminated ovine milk. This study also describes the application of IMC-PCR method to test for M. agalactiae in 516 milk samples collected from sheep with suspected contagious agalactia. Its performance was evaluated relative to culture. This report has demonstrated for the first time, the effective use of rapid and reliable IMC combined with PCR assay for the detection of viable M. agalactiae. The method IMC-PCR provides an alternative to conventional microbiological detection, method and it could be applied to quick detection of M. agalactiae in routine sheep milk samples. © 2014 The Authors published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.

  5. Non-Breeding Eusocial Mole-Rats Produce Viable Sperm--Spermiogram and Functional Testicular Morphology of Fukomys anselli.

    Directory of Open Access Journals (Sweden)

    Angelica Garcia Montero

    Full Text Available Ansell's mole-rats (Fukomys anselli are subterranean rodents living in families composed of about 20 members with a single breeding pair and their non-breeding offspring. Most of them remain with their parents for their lifetime and help to maintain and defend the natal burrow system, forage, and care for younger siblings. Since incest avoidance is based on individual recognition (and not on social suppression we expect that non-breeders produce viable sperm spontaneously. We compared the sperm of breeding and non-breeding males, obtained by electroejaculation and found no significant differences in sperm parameters between both groups. Here, we used electroejaculation to obtain semen for the first time in a subterranean mammal. Spermiogram analysis revealed no significant differences in sperm parameters between breeders and non-breeders. We found significantly larger testes (measured on autopsies and on living animals per ultrasonography of breeders compared to non-breeders (with body mass having a significant effect. There were no marked histological differences between breeding and non-breeding males, and the relative area occupied by Leydig cells and seminiferous tubules on histological sections, respectively, was not significantly different between both groups. The seminiferous epithelium and to a lesser degree the interstitial testicular tissue are characterized by lesions (vacuolar degenerations, however, this feature does not hinder fertilization even in advanced stages of life. The continuous production of viable sperm also in sexually abstinent non-breeders might be best understood in light of the mating and social system of Fukomys anselli, and the potential to found a new family following an unpredictable and rare encounter with an unfamiliar female ("provoked or induced dispersal". Apparently, the non-breeders do not reproduce because they do not copulate but not because they would be physiologically infertile. The significantly

  6. Non-Breeding Eusocial Mole-Rats Produce Viable Sperm—Spermiogram and Functional Testicular Morphology of Fukomys anselli

    Science.gov (United States)

    Garcia Montero, Angelica; Vole, Christiane; Burda, Hynek; Malkemper, Erich Pascal; Holtze, Susanne; Morhart, Michaela; Saragusty, Joseph; Hildebrandt, Thomas B.; Begall, Sabine

    2016-01-01

    Ansell’s mole-rats (Fukomys anselli) are subterranean rodents living in families composed of about 20 members with a single breeding pair and their non-breeding offspring. Most of them remain with their parents for their lifetime and help to maintain and defend the natal burrow system, forage, and care for younger siblings. Since incest avoidance is based on individual recognition (and not on social suppression) we expect that non-breeders produce viable sperm spontaneously. We compared the sperm of breeding and non-breeding males, obtained by electroejaculation and found no significant differences in sperm parameters between both groups. Here, we used electroejaculation to obtain semen for the first time in a subterranean mammal. Spermiogram analysis revealed no significant differences in sperm parameters between breeders and non-breeders. We found significantly larger testes (measured on autopsies and on living animals per ultrasonography) of breeders compared to non-breeders (with body mass having a significant effect). There were no marked histological differences between breeding and non-breeding males, and the relative area occupied by Leydig cells and seminiferous tubules on histological sections, respectively, was not significantly different between both groups. The seminiferous epithelium and to a lesser degree the interstitial testicular tissue are characterized by lesions (vacuolar degenerations), however, this feature does not hinder fertilization even in advanced stages of life. The continuous production of viable sperm also in sexually abstinent non-breeders might be best understood in light of the mating and social system of Fukomys anselli, and the potential to found a new family following an unpredictable and rare encounter with an unfamiliar female (“provoked or induced dispersal”). Apparently, the non-breeders do not reproduce because they do not copulate but not because they would be physiologically infertile. The significantly increased

  7. Action and gait recognition from recovered 3-D human joints.

    Science.gov (United States)

    Gu, Junxia; Ding, Xiaoqing; Wang, Shengjin; Wu, Youshou

    2010-08-01

    A common viewpoint-free framework that fuses pose recovery and classification for action and gait recognition is presented in this paper. First, a markerless pose recovery method is adopted to automatically capture the 3-D human joint and pose parameter sequences from volume data. Second, multiple configuration features (combination of joints) and movement features (position, orientation, and height of the body) are extracted from the recovered 3-D human joint and pose parameter sequences. A hidden Markov model (HMM) and an exemplar-based HMM are then used to model the movement features and configuration features, respectively. Finally, actions are classified by a hierarchical classifier that fuses the movement features and the configuration features, and persons are recognized from their gait sequences with the configuration features. The effectiveness of the proposed approach is demonstrated with experiments on the Institut National de Recherche en Informatique et Automatique Xmas Motion Acquisition Sequences data set.

  8. Dietary strategies to recover from exercise-induced muscle damage.

    Science.gov (United States)

    Sousa, Mónica; Teixeira, Vítor H; Soares, José

    2014-03-01

    Exhaustive or unaccustomed intense exercise can cause exercise-induced muscle damage (EIMD) and its undesirable consequences may decrease the ability to exercise and to adhere to a training programme. This review briefly summarises the muscle damage process, focusing predominantly on oxidative stress and inflammation as contributing factors, and describes how nutrition may be positively used to recover from EIMD. The combined intake of carbohydrates and proteins and the use of antioxidants and/or anti-inflammatory nutrients within physiological ranges are interventions that may assist the recovery process. Although the works studying food instead of nutritional supplements are very scarce, their results seem to indicate that food might be a favourable option as a recovery strategy. To date, the only tested foods were milk, cherries, blueberries and pomegranate with promising results. Other potential solutions are foods rich in protein, carbohydrates, antioxidants and/or anti-inflammatory nutrients.

  9. An experimental study on recovering heat from domestic drain water

    Science.gov (United States)

    Ramadan, Mohamad; Al Shaer, Ali; Haddad, Ahmad; Khaled, Mahmoud

    2016-07-01

    This paper concerns an experimental study on a system of heat recovery applied to domestic drain water pipes. The concept suggested consists of using the heat still present in the drain water as a preheating/heating source to the cold water supply of the building. To proceed, an appropriate experimental setup is developed and a coil heat exchanger is used as heat transfer device in the recovery system. Several scenarios are simulated and corresponding parameters are recorded and analyzed. It was shown that the suggested recovery concept can considerably preheat the cold water supply and then decrease the energy consumption. Particularly, up to 8.6 kW of heat were recovered when the cold water supply is initially at 3 °C.

  10. A computer-implemented method of recovering a visual event

    DEFF Research Database (Denmark)

    2017-01-01

    a synchronization marker at least for a first occurrence of a temporal section classified as a smooth pursuit eye movement; wherein the synchronization marker comprises a link to or impression information of the contents of the viewport at the point in time when the first occurrence of a smooth pursuit eye movement...... of the viewport is displayed, recording an eye movement signal that is indicative of the movements of a user's at least one eye, classifying temporal sections of the eye movement signal into at least a class of long slow-phase OKN eye movements occurring among short slow-phase eye movements; setting...... occurred; via the synchronization marker, recovering the impression information or the contents of the viewport that was displayed at the point in time when the first occurrence of the smooth pursuit occurred....

  11. Cortical neuroplasticity in patients recovering from acute optic neuritis

    DEFF Research Database (Denmark)

    Korsholm, Kirsten; Madsen, Kristoffer Hougaard; Frederiksen, Jette L.

    2008-01-01

    Patients with optic neuritis (ON) undergo cortical and subcortical neuroplasticity as revealed by functional magnetic resonance imaging (fMRI). However, the effect of the heterogeneity of scotomas his not been adequately addressed previously. We introduce a new method of modelling scotomas in f......MRI, to reveal a clearer pattern of neuroplasticity, across a mixed patient population. A longitudinal fMRI-study of visual function in 19 ON patients examined at four timepoints between presentation and 6 months was performed. Four different models were compared. The first model included the four different...... Cortical neuroplasticity during recovery. Moreover, inferences from the fourth model can be extended to the general Population of patients recovering from ON. In Conclusion, we present a method of accommodating subject-specific differences between patients with acute ON by inclusion of an HMD...

  12. Vulnerable GPU Memory Management: Towards Recovering Raw Data from GPU

    Directory of Open Access Journals (Sweden)

    Zhou Zhe

    2017-04-01

    Full Text Available According to previous reports, information could be leaked from GPU memory; however, the security implications of such a threat were mostly over-looked, because only limited information could be indirectly extracted through side-channel attacks. In this paper, we propose a novel algorithm for recovering raw data directly from the GPU memory residues of many popular applications such as Google Chrome and Adobe PDF reader. Our algorithm enables harvesting highly sensitive information including credit card numbers and email contents from GPU memory residues. Evaluation results also indicate that nearly all GPU-accelerated applications are vulnerable to such attacks, and adversaries can launch attacks without requiring any special privileges both on traditional multi-user operating systems, and emerging cloud computing scenarios.

  13. Agglomeration and Deposition Behaviour of Solid Recovered Fuel

    DEFF Research Database (Denmark)

    Pedersen, Morten Nedergaard; Jensen, Peter Arendt; Hjuler, Klaus

    2016-01-01

    Waste-derived fuels, such as solid recovered fuel (SRF), are increasingly being used in, e.g., the cement industry as a means to reduce cost. The inhomogeneous nature of SRF makes it difficult to combust, and many problems may arise within, e.g., combustion control, feeding of fuel, deposit...... formation, or accumulation of impurities. The combustion of polyethylene (PE), polypropylene (PP), polyethylene terephthalate (PET), wood, and SRF were studied in a rotary drum furnace. The combustion was recorded on a camera (60 frames per second), so that any agglomeration or deposition of fuel or ash...... could be monitored. PE and PP pose no significant risk of forming deposits in a combustion environment (T > 800 °C) as a result of a rapid devolatilization, while PET may cause deposits as a result of a sticky char residue. The deposition tendency of the investigated SRF is low, and it may be managed...

  14. Recovering prehistoric woodworking skills using spatial analysis techniques

    Science.gov (United States)

    Kovács, K.; Hanke, K.

    2015-08-01

    Recovering of ancient woodworking skills can be achieved by the simultaneous documentation and analysis of the tangible evidences such as the geometry parameters of prehistoric hand tools or the fine morphological characteristics of well preserved wooden archaeological finds. During this study, altogether 10 different hand tool forms and over 60 hand tool impressions were investigated for the better understanding of the Bronze Age woodworking efficiency. Two archaeological experiments were also designed in this methodology and unknown prehistoric adzes could be reconstructed by the results of these studies and by the spatial analysis of the Bronze Age tool marks. Finally, the trimming efficiency of these objects were also implied and these woodworking skills could be quantified in the case of a Bronze Age wooden construction from Austria. The proposed GIS-based tool mark segmentation and comparison can offer an objective, user-independent technique for the related intangible heritage interpretations in the future.

  15. Recovering geography from a matrix of genetic distances

    Science.gov (United States)

    Serva, M.; Vergni, D.; Volchenkov, D.; Vulpiani, A.

    2017-05-01

    Given a population of N elements with their geographical positions and the genetic (or lexical) distances between couples of elements (inferred, for example, from lexical differences between dialects which are spoken in different towns or from genetic differences between animal populations living in different faunal areas) a very interesting problem is to reconstruct the geographical positions of individuals using only genetic/lexical distances. From a technical point of view the program consists in extracting from the genetic/lexical distances a set of reconstructed geographical positions to be compared with the real ones. We show that geographical recovering is successful when the genetic/lexical distances are not a simple consequence of phylogenesis but also of horizontal transfers as, for example, vocabulary borrowings between different languages. Our results go well beyond the simple observation that geographical distances and genetic/lexical distances are correlated. The ascertainment of a correlation, in our perspective, merely is a prerequisite.

  16. Advances in the recovering of spent lithium battery compounds

    Science.gov (United States)

    Castillo, S.; Ansart, F.; Laberty-Robert, C.; Portal, J.

    Advances in a process based on simple and environmentally compatible operations, aimed to the treatment and recycling of spent lithium-ion batteries, are reported in this paper. This process is safe, economic, and recovers as much of the battery materials as possible. It operates mainly in a selective dissolution in dilute acid, a chemical treatment of the filtrate and a thermal treatment of the solid residue. The validity of the process and its reproducibility have been evaluated at each step of the separation. This method involves very simple equipment and can be scaled-up for commercial production. Based on the projected quantities of lithium ion batteries available for recycling in the next few years, there is a significant market opportunity for a successful technology.

  17. Recovering depth from focus using iterative image estimation techniques

    Energy Technology Data Exchange (ETDEWEB)

    Vitria, J.; Llacer, J.

    1993-09-01

    In this report we examine the possibility of using linear and nonlinear image estimation techniques to build a depth map of a three dimensional scene from a sequence of partially focused images. In particular, the techniques proposed to solve the problem of construction of a depth map are: (1) linear methods based on regularization procedures and (2) nonlinear methods based on statistical modeling. In the first case, we have implemented a matrix-oriented method to recover the point spread function (PSF) of a sequence of partially defocused images. In the second case, the chosen method has been a procedure based on image estimation by means of the EM algorithm, a well known technique in image reconstruction in medical applications. This method has been generalized to deal with optically defocused image sequences.

  18. Swedish recovered wood waste: linking regulation and contamination.

    Science.gov (United States)

    Krook, J; Mårtensson, A; Eklund, M; Libiseller, C

    2008-01-01

    In Sweden, large amounts of wood waste are generated annually from construction and demolition activities, but also from other discarded products such as packaging and furniture. A large share of this waste is today recovered and used for heat production. However, previous research has found that recovered wood waste (RWW) contains hazardous substances, which has significant implications for the environmental performance of recycling. Improved sorting is often suggested as a proper strategy to decrease such implications. In this study, we aim to analyse the impacts of waste regulation on the contamination of RWW. The occurrence of industrial preservative-treated wood, which contains several hazardous substances, was used as an indicator for contamination. First the management of RWW during 1995-2004 was studied through interviews with involved actors. We then determined the occurrence of industrial preservative-treated wood in RWW for that time period for each supplier (actor). From the results, it can be concluded that a substantially less contaminated RWW today relies on extensive source separation. The good news is that some actors, despite several obstacles for such upstream efforts, have already today proved capable of achieving relatively efficient separation. In most cases, however, the existing waste regulation has not succeeded in establishing strong enough incentives for less contaminated waste in general, nor for extensive source separation in particular. One important factor for this outcome is that the current market forces encourage involved actors to practice weak quality requirements and to rely on end-of-pipe solutions, rather than put pressure for improvements on upstream actors. Another important reason is that there is a lack of communication and oversight of existing waste regulations. Without such steering mechanisms, the inherent pressure from regulations becomes neutralized.

  19. Innovative Elution Processes for Recovering Uranium from Seawater

    Energy Technology Data Exchange (ETDEWEB)

    Wai, Chien [Univ. of Idaho, Moscow, ID (United States); Tian, Guoxin [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Janke, Christopher [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2014-05-29

    Utilizing amidoxime-based polymer sorbents for extraction of uranium from seawater has attracted considerable interest in recent years. Uranium collected in the sorbent is recovered typically by elution with an acid. One drawback of acid elution is deterioration of the sorbent which is a significant factor that limits the economic competitiveness of the amidoxime-based sorbent systems for sequestering uranium from seawater. Developing innovative elution processes to improve efficiency and to minimize loss of sorbent capacity become essential in order to make this technology economically feasible for large-scale industrial applications. This project has evaluated several elution processes including acid elution, carbonate elution, and supercritical fluid elution for recovering uranium from amidoxime-based polymer sorbents. The elution efficiency, durability and sorbent regeneration for repeated uranium adsorption- desorption cycles in simulated seawater have been studied. Spectroscopic techniques are used to evaluate chemical nature of the sorbent before and after elution. A sodium carbonate-hydrogen peroxide elution process for effective removal of uranium from amidoxime-based sorbent is developed. The cause of this sodium carbonate and hydrogen peroxide synergistic leaching of uranium from amidoxime-based sorbent is attributed to the formation of an extremely stable uranyl peroxo-carbonato complex. The efficiency of uranium elution by the carbonate-hydrogen peroxide method is comparable to that of the hydrochloric acid elution but damage to the sorbent material is much less for the former. The carbonate- hydrogen peroxide elution also does not need any elaborate step to regenerate the sorbent as those required for hydrochloric acid leaching. Several CO2-soluble ligands have been tested for extraction of uranium from the sorbent in supercritical fluid carbon dioxide. A mixture of hexafluoroacetylacetone and tri-n-butylphosphate shows the best result but uranium

  20. A new non-destructive readout by using photo-recovered surface potential contrast.

    Science.gov (United States)

    Wang, Le; Jin, Kui-juan; Gu, Jun-xing; Ma, Chao; He, Xu; Zhang, Jiandi; Wang, Can; Feng, Yu; Wan, Qian; Shi, Jin-an; Gu, Lin; He, Meng; Lu, Hui-bin; Yang, Guo-zhen

    2014-11-10

    Ferroelectric random access memory is still challenging in the feature of combination of room temperature stability, non-destructive readout and high intensity storage. As a non-contact and non-destructive information readout method, surface potential has never been paid enough attention because of the unavoidable decay of the surface potential contrast between oppositely polarized domains. That is mainly due to the recombination of the surface movable charges around the domain walls. Here, by introducing a laser beam into the combination of piezoresponse force microscopy and Kelvin probe force microscopy, we demonstrate that the surface potential contrast of BiFeO3 films can be recovered under light illumination. The recovering mechanism is understood based on the redistribution of the photo-induced charges driven by the internal electric field. Furthermore, we have created a 12-cell memory pattern based on BiFeO3 films to show the feasibility of such photo-assisted non-volatile and non-destructive readout of the ferroelectric memory.

  1. Sustainable polysaccharide-based biomaterial recovered from waste aerobic granular sludge as a surface coating material

    NARCIS (Netherlands)

    Lin, Y. M.; Nierop, K.G.J.|info:eu-repo/dai/nl/182329895; Girbal-Neuhauser, E.; Adriaanse, M.; van Loosdrecht, M. C M

    To evaluate the possibility of utilizing polysaccharide-based biomaterial recovered from aerobic granular sludge as a coating material, the morphology, molecular weight distribution and chemical composition of the recovered biomaterial were investigated by atomic force microscopy, size exclusion

  2. Sustainable polysaccharide-based biomaterial recovered from waste aerobic granular sludge as a surface coating material

    NARCIS (Netherlands)

    Lin, Y.M.; Nierop, K.G.J.; Girbal-Neuhauser, E.; Adriaanse, M.; Van Loosdrecht, M.C.M.

    2015-01-01

    To evaluate the possibility of utilizing polysaccharide-based biomaterial recovered from aerobic granular sludge as a coating material, the morphology, molecular weight distribution and chemical composition of the recovered biomaterial were investigated by atomic force microscopy, size exclusion

  3. Visualization and 3D reconstruction of flame cells of Taenia solium (cestoda.

    Directory of Open Access Journals (Sweden)

    Laura E Valverde-Islas

    Full Text Available BACKGROUND: Flame cells are the terminal cells of protonephridial systems, which are part of the excretory systems of invertebrates. Although the knowledge of their biological role is incomplete, there is a consensus that these cells perform excretion/secretion activities. It has been suggested that the flame cells participate in the maintenance of the osmotic environment that the cestodes require to live inside their hosts. In live Platyhelminthes, by light microscopy, the cells appear beating their flames rapidly and, at the ultrastructural, the cells have a large body enclosing a tuft of cilia. Few studies have been performed to define the localization of the cytoskeletal proteins of these cells, and it is unclear how these proteins are involved in cell function. METHODOLOGY/PRINCIPAL FINDINGS: Parasites of two different developmental stages of T. solium were used: cysticerci recovered from naturally infected pigs and intestinal adults obtained from immunosuppressed and experimentally infected golden hamsters. Hamsters were fed viable cysticerci to recover adult parasites after one month of infection. In the present studies focusing on flame cells of cysticerci tissues was performed. Using several methods such as video, confocal and electron microscopy, in addition to computational analysis for reconstruction and modeling, we have provided a 3D visual rendition of the cytoskeletal architecture of Taenia solium flame cells. CONCLUSIONS/SIGNIFICANCE: We consider that visual representations of cells open a new way for understanding the role of these cells in the excretory systems of Platyhelminths. After reconstruction, the observation of high resolution 3D images allowed for virtual observation of the interior composition of cells. A combination of microscopic images, computational reconstructions and 3D modeling of cells appears to be useful for inferring the cellular dynamics of the flame cell cytoskeleton.

  4. Visualization and 3D Reconstruction of Flame Cells of Taenia solium (Cestoda)

    Science.gov (United States)

    Valverde-Islas, Laura E.; Arrangoiz, Esteban; Vega, Elio; Robert, Lilia; Villanueva, Rafael; Reynoso-Ducoing, Olivia; Willms, Kaethe; Zepeda-Rodríguez, Armando; Fortoul, Teresa I.; Ambrosio, Javier R.

    2011-01-01

    Background Flame cells are the terminal cells of protonephridial systems, which are part of the excretory systems of invertebrates. Although the knowledge of their biological role is incomplete, there is a consensus that these cells perform excretion/secretion activities. It has been suggested that the flame cells participate in the maintenance of the osmotic environment that the cestodes require to live inside their hosts. In live Platyhelminthes, by light microscopy, the cells appear beating their flames rapidly and, at the ultrastructural, the cells have a large body enclosing a tuft of cilia. Few studies have been performed to define the localization of the cytoskeletal proteins of these cells, and it is unclear how these proteins are involved in cell function. Methodology/Principal Findings Parasites of two different developmental stages of T. solium were used: cysticerci recovered from naturally infected pigs and intestinal adults obtained from immunosuppressed and experimentally infected golden hamsters. Hamsters were fed viable cysticerci to recover adult parasites after one month of infection. In the present studies focusing on flame cells of cysticerci tissues was performed. Using several methods such as video, confocal and electron microscopy, in addition to computational analysis for reconstruction and modeling, we have provided a 3D visual rendition of the cytoskeletal architecture of Taenia solium flame cells. Conclusions/Significance We consider that visual representations of cells open a new way for understanding the role of these cells in the excretory systems of Platyhelminths. After reconstruction, the observation of high resolution 3D images allowed for virtual observation of the interior composition of cells. A combination of microscopic images, computational reconstructions and 3D modeling of cells appears to be useful for inferring the cellular dynamics of the flame cell cytoskeleton. PMID:21412407

  5. Effects of orally administered viable Lactobacillus rhamnosus GG and Propionibacterium freudenreichii subsp. shermanii JS on mouse lymphocyte proliferation.

    Science.gov (United States)

    Kirjavainen, P V; ElNezami, H S; Salminen, S J; Ahokas, J T; Wright, P F

    1999-11-01

    Immunomodulation by probiotics is a subject of growing interest, but the knowledge of dose response and time profile relationships is minimal. In this study we examined the effects of Lactobacillus rhamnosus GG (LGG) and Propionibacterium freudenreichii subsp. shermanii JS (PJS) on the proliferative activity of murine lymphocytes ex vivo. Dose dependency was assessed by treating animals perorally with a low or a high dose (i.e., 10(9) or 10(12) viable bacteria/kg of body weight) for 7 days. The lower dose levels of each strain appeared to enhance T-cell proliferation at the optimal concanavalin A (ConA) concentration (by 69 to 84%) and B-cell proliferation at the optimal and supraoptimal concentrations of lipopolysaccharide (by 57 to 82%). B-cell proliferation was also enhanced by the high LGG dose (by 32 to 39%) but was accompanied by a marginal decrease in T-cell proliferation (by 8%) at the optimal ConA concentration. The time profiles of the immune responses were assessed after daily treatment with the higher dose for 3, 7, and 14 days. A significant decrease in basal lymphoproliferation (by 32 to 42%) was observed with PJS treatment after the 3- and 7-day periods; however, this activity returned to control levels after 14 days of treatment, which also resulted in significantly enhanced T-cell proliferation at optimal and supraoptimal ConA concentrations (by 24 to 80%). The 14-day LGG treatment also enhanced the latter activity (by 119%). In conclusion, LGG and PJS have specific dose- and duration-dependent immunomodulatory effects on the proliferative activity of B and T lymphocytes and may also reduce lymphocyte sensitivity to the cytotoxic effects of lectin mitogens.

  6. Use of propidium monoazide and quantitative PCR for differentiation of viable Escherichia coli from E. coli killed by mild or pasteurizing heat treatments.

    Science.gov (United States)

    Yang, Xianqin; Badoni, Madhu; Gill, Colin O

    2011-12-01

    Suspensions of Escherichia coli in peptone water were heated at temperatures between 52 and 90 °C, inclusive. Samples withdrawn at suitable times were not or were treated with propidium monoazide (PMA) or deoxycholate then PMA before extraction of DNA. DNA was quantified by real-time PCR for estimation of the numbers of E. coli from which template DNA for the PCR was obtained. Numbers of viable E. coli in suspensions at the times of sampling were determined from plate counts. For samples from suspensions heated at temperatures ≥ 52 ≤ 72 °C, PCR cycle threshold (Ct) values were little or no different for DNA from corresponding samples that were or were not treated with PMA. PMA treatment of samples heated to ≥ 80 °C largely inactivated E. coli DNA for PCR. When samples heated to ≤ 72 °C were treated with deoxycholate before treatment with PMA, Ct values for treated samples were greater than the Ct values for the corresponding untreated samples. Similar results were obtained with E. coli suspended in milk or fluid from ground beef pummeled with diluent. The results indicate that cells killed by heating to ≥ 80 °C are permeable to PMA, but most cells killed by heating to ≤ 72 °C are not. However, treatment with deoxycholate renders a substantial fraction of the latter cells permeable to PMA. Numbers of viable or dead E. coli can then be estimated from Ct values for samples not treated or treated with deoxycholate and PMA, provided viable cells are ≥ 1% of the total. Copyright © 2011 Elsevier Ltd. All rights reserved.

  7. Isolation of viable Neospora caninum from brains of wild gray wolves (Canis lupus).

    Science.gov (United States)

    Dubey, J P; Jenkins, M C; Ferreira, L R; Choudhary, S; Verma, S K; Kwok, O C H; Fetterer, R; Butler, E; Carstensen, M

    2014-03-17

    Neospora caninum is a common cause of abortion in cattle worldwide. Canids, including the dog and the dingo (Canis familiaris), the coyote (Canis latrans), and the gray wolf (Canis lupus) are its definitive hosts that can excrete environmentally resistant oocysts in the environment, but also can act as intermediate hosts, harboring tissue stages of the parasite. In an attempt to isolate viable N. caninum from tissues of naturally infected wolves, brain and heart tissue from 109 wolves from Minnesota were bioassayed in mice. Viable N. caninum (NcWolfMn1, NcWolfMn2) was isolated from the brains of two wolves by bioassays in interferon gamma gene knockout mice. DNA obtained from culture-derived N. caninum tachyzoites of the two isolates were analyzed by N. caninum-specific Nc5 polymerase chain reaction and confirmed diagnosis. This is the first report of isolation of N. caninum from tissues of any wild canid host. Published by Elsevier B.V.

  8. Viable Intrauterine Pregnancy and Coexisting Molar Pregnancy in a Bicornuate Uterus: A Rare Presentation

    Directory of Open Access Journals (Sweden)

    Kavitha Krishnamoorthy

    2016-01-01

    Full Text Available A complete hydatidiform mole with a viable coexisting fetus (CMCF is a rare occurrence. Similarly, Mullerian anomalies such as a bicornuate uterus are uncommon variants of normal anatomy. We report a case of a 40-year-old female with a known bicornuate uterus presenting at 13 weeks gestation with vaginal bleeding. Ultrasound findings showed a healthy viable pregnancy in the right horn with complete molar pregnancy in the left horn. After extensive counseling, the patient desired conservative management, however, was unable to continue due to profuse vaginal bleeding. The patient underwent suction dilation and curettage under general anesthesia and evacuation of the uterine horns. Postoperatively, the patient was followed until serum beta-human chorionic gonadotropin (β-hCG level dropped to <5 mU. This is the first case of a CMCF reported in a bicornuate uterus, diagnosed with the use of ultrasound imaging.

  9. Optoelectrofluidic printing system for fabricating hydrogel sheets with on-demand patterned cells and microparticles.

    Science.gov (United States)

    Gi, Hyun Ji; Han, Dongsik; Park, Je-Kyun

    2017-01-16

    This paper presents a novel optoelectrofluidic printing system that facilitates not only the optoelectrofluidic patterning of microparticles and mammalian cells but also the harvesting of the patterned microparticles encapsulated within poly(ethylene glycol) dicarylate (PEGDA) hydrogel sheets. Although optoelectrofluidic technology has numerous advantages for programmable and on-demand patterning and the feasibility of manipulating single microparticles, practical applications using existing laboratory infrastructure in biological and clinical research fields have been strictly restricted due to the impossibility of recovering the final patterned product. In order to address these harvesting limitations, PEGDA was employed to utilize optoelectrofluidic printing. The Clausius-Mossotti factor was calculated to investigate the dielectrophoretic mobility of the microparticle and the cell in the PEGDA precursor solution. As a proof of concept, three basic controllabilities of the optoelectrofluidic printing system were characterized: the number of microparticles, the distance between the microparticle columns, and the ratio of two different microparticles. Furthermore, the optoelectrofluidic patterning and printing of human liver carcinoma cells (HepG2) were demonstrated in 5 min with a single-cell level of resolution. The appropriate ranges of frequency were experimentally defined based on the calculated result of the dielectrophoretic mobility of HepG2 cells. Finally, optoelectrofluidically cell-patterned hydrogel sheets were successfully recovered under a highly viable physiological condition.

  10. Improved identification of viable myocardium using second harmonic imaging during dobutamine stress echocardiography

    OpenAIRE

    Sozzi, Fabiola; Poldermans, Don; Bax, Jeroen; Elhendy, Abdou; Vourvouri, Eleni; Valkema, Roelf; Sutter, J.; Schinkel, Arend; Borghetti, A; Roelandt, Jos

    2001-01-01

    OBJECTIVE—To determine whether, compared with fundamental imaging, second harmonic imaging can improve the accuracy of dobutamine stress echocardiography for identifying viable myocardium, using nuclear imaging as a reference.
PATIENTS—30 patients with chronic left ventricular dysfunction (mean (SD) age, 60 (8) years; 22 men).
METHODS—Dobutamine stress echocardiography was carried out in all patients using both fundamental and second harmonic imaging. All patients underwent dual isotope simul...

  11. Cosmic constraint on massive neutrinos in viable f( R) gravity with producing Λ CDM background expansion

    Science.gov (United States)

    Lu, Jianbo; Liu, Molin; Wu, Yabo; Wang, Yan; Yang, Weiqiang

    2016-12-01

    Tensions between several cosmic observations were found recently, such as the inconsistent values of H0 (or σ 8) were indicated by the different cosmic observations. Introducing the massive neutrinos in Λ CDM could potentially solve the tensions. Viable f( R) gravity producing Λ CDM background expansion with massive neutrinos is investigated in this paper. We fit the current observational data: Planck-2015 CMB, RSD, BAO, and SNIa to constrain the mass of neutrinos in viable f( R) theory. The constraint results at 95% confidence level are: Σ m_ν case, m_{ν , sterile}^effcase. For the effects due to the mass of the neutrinos, the constraint results on model parameter at 95% confidence level become f_{R0}× 10^{-6}> -1.89 and f_{R0}× 10^{-6}> -2.02 for two cases, respectively. It is also shown that the fitting values of several parameters much depend on the neutrino properties, such as the cold dark matter density, the cosmological quantities at matter-radiation equality, the neutrino density and the fraction of baryonic mass in helium. Finally, the constraint result shows that the tension between direct and CMB measurements of H_0 gets slightly weaker in the viable f( R) model than that in the base Λ CDM model.

  12. Improved identification of viable myocardium using second harmonic imaging during dobutamine stress echocardiography.

    Science.gov (United States)

    Sozzi, F B; Poldermans, D; Bax, J J; Elhendy, A; Vourvouri, E C; Valkema, R; De Sutter, J; Schinkel, A F; Borghetti, A; Roelandt, J R

    2001-12-01

    To determine whether, compared with fundamental imaging, second harmonic imaging can improve the accuracy of dobutamine stress echocardiography for identifying viable myocardium, using nuclear imaging as a reference. 30 patients with chronic left ventricular dysfunction (mean (SD) age, 60 (8) years; 22 men). Dobutamine stress echocardiography was carried out in all patients using both fundamental and second harmonic imaging. All patients underwent dual isotope simultaneous acquisition single photon emission computed tomography (DISA-SPECT) with (99m)technetium-tetrofosmin/(18)F-fluorodeoxyglucose on a separate day. Myocardial viability was considered present by dobutamine stress echocardiography when segments with severe dysfunction showed a biphasic sustained improvement or an ischaemic response. Viability criteria on DISA-SPECT were normal or mildly reduced perfusion and metabolism, or perfusion/metabolism mismatch. Using fundamental imaging, 330 segments showed severe dysfunction at baseline; 144 (44%) were considered viable. The agreement between dobutamine stress echocardiography by fundamental imaging and DISA-SPECT was 78%, kappa = 0.56. Using second harmonic imaging, 288 segments showed severe dysfunction; 138 (48%) were viable. The agreement between dobutamine stress echocardiography and DISA-SPECT was significantly better when second harmonic imaging was used (89%, kappa = 0.77, p = 0.001 v fundamental imaging). Second harmonic imaging applied during dobutamine stress echocardiography increases the agreement with DISA-SPECT for detecting myocardial viability.

  13. The dose-fractionation sensitivity of the kidney; assessment of viable tubule cross-sections at 19 months after X irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Jen, Y.-M.; Hendry, J.H. (Christie Hospital and Holt Radium Inst., Manchester (United Kingdom). Paterson Labs.)

    1993-03-01

    The formation of viable tubule cross-sections was assessed in histological sections of murine kidneys at 19 months after fractionated bilateral X-ray doses with 12 h intervals between fractions. The data were analysed using the linear-quadratic model which provides values of [alpha] and [beta] characterizing the slope of the dose-response curve, and the ratio of [alpha] and [beta] indicative of the sparing effect of dose fractionation. The tubule data were characterized by [alpha] = 0.057 [+-] 0.009 Gy[sup -1], [beta] = 0.011 [+-] 0.001 Gy[sup -2], [alpha]/[beta] = 5.0 [+-] 0.9 Gy. Also, the number of cells (per focus region of the nephron) calculated as being capable of producing a viable focus was 2.5 [+-] 0.5, which was confirmed using a separate two-dose approach (2.1 [+-] 0.3). Together with other data, of the order of 1000 regenerative cells per nephron (10[sup 4] total cells) can be deduced. The values of the fractionation sensitivity parameters are similar to values measured previously for cells taken from irradiated kidneys up to a year after irradiation and forming colonies in primary culture, and also similar to values assessed using various functional measures of kidney injury. (Author).

  14. Existence of both culturable and viable but non culturable (VNC) E. coli populations with distinct settling velocities in karst aquifer

    Science.gov (United States)

    Petit, F.; Ratajczak, M.; Massei, N.; Lafite, R.; Clermont, O.; Denamur, E.; Berthe, T.

    2012-12-01

    associated to "non settleable" particles, decreased from the sinkhole to the spring. In contrast in dry period while the turbidity and the contamination by culturable E. coli is low, the three populations of viable, culturable and VNC E. coli are introduced in the karst. Within the population of culturable E. coli, persistent strains mainly belonging to B1 phylogroup and growing at 7°C were introduced in karst aquifer. At the spring, whatever the class of particles up to 96% of viable cells were in VNC state suggesting that a physiological change occurred within aquifer, independently of the association of E. coli with particles. At the well, the pumping of the water induced an input of resident VNC E. coli consecutively to a resuspension of particles previously settled within the karstic network, during a past turbid event.

  15. USE OF POLYMERS TO RECOVER VISCOUS OIL FROM UNCONVENTIONAL RESERVOIRS

    Energy Technology Data Exchange (ETDEWEB)

    Randall Seright

    2011-09-30

    This final technical progress report summarizes work performed the project, 'Use of Polymers to Recover Viscous Oil from Unconventional Reservoirs.' The objective of this three-year research project was to develop methods using water soluble polymers to recover viscous oil from unconventional reservoirs (i.e., on Alaska's North Slope). The project had three technical tasks. First, limits were re-examined and redefined for where polymer flooding technology can be applied with respect to unfavorable displacements. Second, we tested existing and new polymers for effective polymer flooding of viscous oil, and we tested newly proposed mechanisms for oil displacement by polymer solutions. Third, we examined novel methods of using polymer gels to improve sweep efficiency during recovery of unconventional viscous oil. This report details work performed during the project. First, using fractional flow calculations, we examined the potential of polymer flooding for recovering viscous oils when the polymer is able to reduce the residual oil saturation to a value less than that of a waterflood. Second, we extensively investigated the rheology in porous media for a new hydrophobic associative polymer. Third, using simulation and analytical studies, we compared oil recovery efficiency for polymer flooding versus in-depth profile modification (i.e., 'Bright Water') as a function of (1) permeability contrast, (2) relative zone thickness, (3) oil viscosity, (4) polymer solution viscosity, (5) polymer or blocking-agent bank size, and (6) relative costs for polymer versus blocking agent. Fourth, we experimentally established how much polymer flooding can reduce the residual oil saturation in an oil-wet core that is saturated with viscous North Slope crude. Finally, an experimental study compared mechanical degradation of an associative polymer with that of a partially hydrolyzed polyacrylamide. Detailed results from the first two years of the project may be

  16. Previously undescribed plasmids recovered from activated sludge confer tetracycline resistance and phenotypic changes to Acinetobacter oleivorans DR1.

    Science.gov (United States)

    Hong, Hyerim; Ko, Hyeok-Jin; Choi, In-Geol; Park, Woojun

    2014-02-01

    We used culture-dependent and culture-independent methods to extract previously undescribed plasmids harboring tetracycline (TC) resistance genes from activated sludge. The extracted plasmids were transformed into naturally competent Acinetobacter oleivorans DR1 to recover a non-Escherichia coli-based plasmid. The transformed cells showed 80-100-fold higher TC resistance than the wild-type strain. Restriction length polymorphism performed using 30 transformed cells showed four different types of plasmids. Illumina-based whole sequencing of the four plasmids identified three previously unreported plasmids and one previously reported plasmid. All plasmids carried TC resistance-related genes (tetL, tetH), tetracycline transcriptional regulators (tetR), and mobilization-related genes. As per expression analysis, TC resistance genes were functional in the presence of TC. The recovered plasmids showed mosaic gene acquisition through horizontal gene transfer. Membrane fluidity, hydrophobicity, biofilm formation, motility, growth rate, sensitivity to stresses, and quorum sensing signals of the transformed cells were different from those of the wild-type cells. Plasmid-bearing cells seemed to have an energy burden for maintaining and expressing plasmid genes. Our data showed that acquisition of TC resistance through plasmid uptake is related to loss of biological fitness. Thus, cells acquiring antibiotic resistance plasmids can survive in the presence of antibiotics, but must pay ecological costs.

  17. Filling of recovered mining areas using solidifying backfill

    Directory of Open Access Journals (Sweden)

    Zeman Róbert

    2001-12-01

    Full Text Available The aim of this article is to explore the possibilities for filling recovered mining areas using solidifying backfill .The article describes the preparation of the backfill (backfill formulation with an eventual application using low quality sands, wastes from treatment plants and ash from power plants etc now to transport it as well as its application in practice. Advantageous and disadvantageous of this method are also mentioned.Several factors must be taken info consideration during the preparation process of the backfill mixture. Firstly, the quantities of each individual component must be constantly regulated. Secondly, the properties of each component must be respected. In addition, the needs of the pipeline transport system and the specific conditions of the recovered area to be filled must also be considered.Hydraulic transport and pneumo-hydraulic pipeline transport are used for handling the backfill. Pumps for transporting the solidifying backfill have to carry out demanding tasks.Due to the physical-mechanical properties of the backfill, only highly powerful pumps can be considered. Piston type pumps such as Abel Simplex and Duplex pumps with capacities of up to 100 m3.h-1 and operating pressures of up to 16 MPa would be suitable.This method has been applied abroad for different purposes. For example, solid backfill was used in the Hamr mine during exploitation of uranium using the room-and-pillar system mining method.In the Ostrava–Karvina Coal field, backfill was used in decontamination work, filling areas in a zone of dangerous deformations and for creating a dividing stratum during thick seam mining.Research info the use of solidifying backfill was also done in the Walsum mine in Germany. The aim of this research was:- to investigate the possibilities of filling a collapsing area in a working face using a solidifying mixture of power plant ash and water,- to verify whether towing pipelines proposed by the DMT corporation would be

  18. Fractional diffusion: recovering the distributed fractional derivative from overposed data

    Science.gov (United States)

    Rundell, W.; Zhang, Z.

    2017-03-01

    There has been considerable recent study in ‘subdiffusion’ models that replace the standard parabolic equation model by a one with a fractional derivative in the time variable. There are many ways to look at this newer approach and one such is to realize that the order of the fractional derivative is related to the time scales of the underlying diffusion process. This raises the question of what order α of derivative should be taken and if a single value actually suffices. This has led to models that combine a finite number of these derivatives each with a different fractional exponent {αk} and different weighting value c k to better model a greater possible range of time scales. Ultimately, one wants to look at a situation that combines derivatives in a continuous way—the so-called distributional model with parameter μ ≤ft(α \\right) . However all of this begs the question of how one determines this ‘order’ of differentiation. Recovering a single fractional value has been an active part of the process from the beginning of fractional diffusion modeling and if this is the only unknown then the markers left by the fractional order derivative are relatively straightforward to determine. In the case of a finite combination of derivatives this becomes much more complex due to the more limited analytic tools available for such equations, but recent progress in this direction has been made, (Li et al 2015 Appl. Math. Comput. 257 381-97, Li and Yamamoto 2015 Appl. Anal. 94 570-9). This paper considers the full distributional model where the order is viewed as a function μ ≤ft(α \\right) on the interval (0, 1]. We show existence, uniqueness and regularity for an initial-boundary value problem including an important representation theorem in the case of a single spatial variable. This is then used in the inverse problem of recovering the distributional coefficient μ ≤ft(α \\right) from a time trace of the solution and a uniqueness result is proven.

  19. Alienation, recovered animism and altered states of consciousness.

    Science.gov (United States)

    Charlton, Bruce G

    2007-01-01

    Alienation is the feeling that life is 'meaningless', that we do not belong in the world. But alienation is not an inevitable part of the human condition: some people do feel at one with the world as a consequence of the animistic way of thinking which is shared by children and hunter-gatherers. Animism considers all significant entities to have 'minds', to be 'alive', to be sentient agents. The animistic thinker inhabits a world populated by personal powers including not just other human beings, but also important animals and plants, and significant aspects of physical landscape. Humans belong in this world because it is a web of social relationships. Animism is therefore spontaneous, the 'natural' way of thinking for humans: all humans began as animistic children and for most of human evolutionary history would have grown into animistic adults. It requires sustained, prolonged and pervasive formal education to 'overwrite' animistic thinking with the rationalistic objectivity typical of the modern world. It is this learned abstraction that creates alienation--humans are no longer embedded in a world of social relations but become estranged, adrift in a world of indifferent things. Methods used to cure alienation and recover animistic modes of thinking involve detachment from the social systems that tend to maintain objectivity and rationality: for example, solitude, leisure, unstructured time and direct contact with nature. Many people also achieve similar results by deliberately inducing altered states of consciousness. Animistic thinking may emerge in meditation or contemplation, lucid dreaming, from self-hypnosis, when drowsy, in 'trance states' induced by repetitious rhythm or light, or when delirious due to illness, brain injury, psychoses, or intoxication with 'entheogenic' drugs--which is probably one reason for the perennial popularity of inducing intoxicated states. However, intoxication will typically damage memory processes making it harder to learn

  20. CHO-K1 host cells adapted to growth in glutamine-free medium by FACS-assisted evolution.

    Science.gov (United States)

    Bort, Juan A Hernández; Stern, Beate; Borth, Nicole

    2010-10-01

    During the process of recombinant cell line optimisation for production of biopharmaceuticals, multiple cellular properties like robustness against stress, the attainment of high cell concentrations and maintenance of high viability must be considered to maximize protein yield. To improve growth and viability, glutamine is supplemented as an alternative energy source for rapidly dividing cells that oxidize glucose inefficiently. However, the resulting by-product ammonia is toxic at high concentrations and has a negative impact on protein glycosylation, a major quality-determining parameter of biopharmaceuticals. In this work, the CHO-K1 cell line was adapted to a chemically defined medium and suspension growth within 3 weeks. Subsequently, the glutamine concentration was stepwise reduced from 8 to 4 and 2 mM. After each reduction, both the final cell concentration in the batch and the viability decreased. To force a rapid evolution of cells to achieve high final cell concentrations, cells were seeded at high densities (10(7) cells/mL) and surviving cells were sorted by FACS or MACS when viability declined to 10% (typically after 24 h). Sorted cells were grown in batch until viability declined to 10% and viable cells recovered again. The final sorted population was able to reach comparable or even better viable cell concentrations and showed a significantly improved viability compared to their ancestors. The 2 mM glutamine-adapted cell line was directly transferred into glutamine-free medium and was able to grow at comparable rates without requiring further adaptation. Cells compensated the lack of glutamine by increasing their consumption of glutamate and aspartate.

  1. Recovering from a bad start: rapid adaptation and tradeoffs to growth below a threshold density

    Directory of Open Access Journals (Sweden)

    Marx Christopher J

    2012-07-01

    Full Text Available Abstract Background Bacterial growth in well-mixed culture is often assumed to be an autonomous process only depending upon the external conditions under control of the investigator. However, increasingly there is awareness that interactions between cells in culture can lead to surprising phenomena such as density-dependence in the initiation of growth. Results Here I report the unexpected discovery of a density threshold for growth of a strain of Methylobacterium extorquens AM1 used to inoculate eight replicate populations that were evolved in methanol. Six of these populations failed to grow to the expected full density during the first couple transfers. Remarkably, the final cell number of six populations crashed to levels 60- to 400-fold smaller than their cohorts. Five of these populations recovered to full density soon after, but one population remained an order of magnitude smaller for over one hundred generations. These variable dynamics appeared to be due to a density threshold for growth that was specific to both this particular ancestral strain and to growth on methanol. When tested at full density, this population had become less fit than its ancestor. Simply increasing the initial dilution 16-fold reversed this result, revealing that this population had more than a 3-fold advantage when tested at this lower density. As this population evolved and ultimately recovered to the same final density range as the other populations this low-density advantage waned. Conclusions These results demonstrate surprisingly strong tradeoffs during adaptation to growth at low absolute densities that manifest over just a 16-fold change in density. Capturing laboratory examples of transitions to and from growth at low density may help us understand the physiological and evolutionary forces that have led to the unusual properties of natural bacteria that have specialized to low-density environments such as the open ocean.

  2. Detection of viable enterotoxin-producing Bacillus cereus and analysis of toxigenicity from ready-to-eat foods and infant formula milk powder by multiplex PCR.

    Science.gov (United States)

    Zhang, Zhihong; Feng, Lixia; Xu, Hengyi; Liu, Chengwei; Shah, Nagendra P; Wei, Hua

    2016-02-01

    Bacillus cereus is responsible for several outbreaks of foodborne diseases due to its emetic toxin and enterotoxin. Enterotoxins, cytotoxin K (CytK), nonhemolytic enterotoxin (Nhe), and hemolysin BL (Hbl), have been recorded in several diarrheal cases due to food poisoning from B. cereus. The objective of this study was to develop a rapid and accurate method that combines multiplex PCR with propidium monoazide to selectively detect viable cells of enterotoxin-producing B. cereus in milk powder, noodles, and rice, and investigate the distribution of enterotoxins in 62 strains of B. cereus in Jiangxi province, China. The specificity of primers of 3 enterotoxins (i.e., cytK, nheA, and hblD) of B. cereus was verified by inclusivity and exclusivity tests using single PCR. Upon optimization of multiplex PCR conditions, it was found that the detection limit of viable cells was 10(2) cfu/mL of B. cereus in pure culture. By enrichment for 3 or 4 h and propidium monoazide pretreatment, a protocol for detection of viable cells as low as 2.2×10(1) cfu/g in spiked food (e.g., milk powder, noodles, and rice) was established and proved valid even under the interference of non-Bacillus cereus at as high as 10(5) cfu/g. Moreover, the protocol based on multiplex PCR for detection was applied for the analysis of distribution of toxin gene of B. cereus, and the results showed a regional feature for toxin gene distribution, indicating that potential toxigenicity of B. cereus should be evaluated further. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  3. Cell structure and percent viability by a slide centrifuge technique.

    Science.gov (United States)

    Fitzgerald, M G; Hosking, C S

    1982-01-01

    It was found that a slide centrifuge (Cytospin) preparation of a cell suspension allowed a reliable assessment of not only cell structure but also the percentage of non-viable cells. The non-viable cells appeared as "smear" cells and paralleled in number the cells taking up trypan blue. Direct experiment showed the unstained viable cells in a trypan blue cell suspension remained intact in a Cytospin preparation while the cells taking up trypan blue were the "smear" cells. The non-viability of the "smear" cells was confirmed by their inability to survive in culture. Images PMID:7040483

  4. CYCLODEXTRINS TO RECOVER TEXTILE DYES IN WASTE WATER

    Directory of Open Access Journals (Sweden)

    FRANCO Esther

    2015-05-01

    Full Text Available Cyclodextrins are cyclic oligosaccharides with a special toroid shape, obtained by the action of glucosyltransferase enzyme (CGTase on starch molecule. Their peculiar structure allows the accommodation of different guest molecules inside their cavity forming molecular inclusion complexes. There are different types depending on the glucose units that are formed, called native. The cyclodextrins can be modified incorporating different groups (hydroxipropyl, methyl... that changes their properties. Due their versatility in size, properties and the variety of inclusion complex can form is employed in many different industries like pharmacy, food or cosmetics to protect the molecule or to reduce their volatility. As the guest molecule is not bond with the cyclodextrin with the appropriate conditions it can release easily. In textile industry had been use in different areas: to remove surfactants from washed textiles, to substitute surfactants, in the dyeing process, in detergents… Due their capacity to fix onto textile allows the functionalization of the fabrics giving them new properties like UV protection, antimicrobial or insect repellents depending on the guest molecule, in. The project DYES4EVER employs the cyclodextrins to encapsulate dyes not fixed during the dye process that remains in the wastewater and aims to go one step further and reuse the dyes recovered as a raw material in new dyeing processes.

  5. Recovering glycoside hydrolase genes from active tundra cellulolytic bacteria.

    Science.gov (United States)

    Pinnell, Lee J; Dunford, Eric; Ronan, Patrick; Hausner, Martina; Neufeld, Josh D

    2014-07-01

    Bacteria responsible for cellulose hydrolysis in situ are poorly understood, largely because of the relatively recent development of cultivation-independent methods for their detection and characterization. This study combined DNA stable-isotope probing (DNA-SIP) and metagenomics for identifying active bacterial communities that assimilated carbon from glucose and cellulose in Arctic tundra microcosms. Following DNA-SIP, bacterial fingerprint analysis of gradient fractions confirmed isotopic enrichment. Sequenced fingerprint bands and clone library analysis of 16S rRNA genes identified active bacterial taxa associated with cellulose-associated labelled DNA, including Bacteroidetes (Sphingobacteriales), Betaproteobacteria (Burkholderiales), Alphaproteobacteria (Caulobacteraceae), and Chloroflexi (Anaerolineaceae). We also compared glycoside hydrolase metagenomic profiles from bulk soil and heavy DNA recovered from DNA-SIP incubations. Active populations consuming [(13)C]glucose and [(13)C]cellulose were distinct, based on ordinations of light and heavy DNA. Metagenomic analysis demonstrated a ∼3-fold increase in the relative abundance of glycoside hydrolases in DNA-SIP libraries over bulk-soil libraries. The data also indicate that multiple displacement amplification introduced bias into the resulting metagenomic analysis. This research identified DNA-SIP incubation conditions for glucose and cellulose that were suitable for Arctic tundra soil and confirmed that DNA-SIP enrichment can increase target gene frequencies in metagenomic libraries.

  6. Biodiesel production from residual oils recovered from spent bleaching earth

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Yi-Pin; Chang, James I. [Department of Safety, Health and Environmental Engineering, National Kaohsiung First University of Science and Technology, 1, University Blvd., Yenchao, Kaohsiung (China)

    2010-01-15

    This work was to study technical and economic feasibilities of converting residual oils recovered from spent bleaching earth generated at soybean oil refineries into useable biodiesel. Experimental results showed that fatty acids in the SBE residual oil were hexadecenoic acid (58.19%), stearic acid (21.49%) and oleic acid (20.32%), which were similar to those of vegetable oils. The methyl ester conversion via a transesterification process gave a yield between 85 and 90%. The biodiesel qualities were in reasonable agreement with both EN 14214 and ASTM D6751 standards. A preliminary financial analysis showed that the production cost of biodiesel from SBE oils was significantly lower than the pre-tax price of fossil diesel or those made of vegetable oils or waste cooking oils. The effects of the crude oil price and the investment on the production cost and the investment return period were also conducted. The result showed that the investment would return faster at higher crude oil price. (author)

  7. Emotional intelligence and recovering from induced negative emotional state

    Science.gov (United States)

    Limonero, Joaquín T.; Fernández-Castro, Jordi; Soler-Oritja, Jordi; Álvarez-Moleiro, María

    2015-01-01

    The aim of the present study was to examine the relationship between emotional intelligence (EI) and recovering from negative emotions induction, using a performance test to measure EI. Sixty seven undergraduates participated in the procedure, which lasted 75 min and was divided into three stages. At Time 1, subjects answered the State-Trait Anxiety Inventory (STAI)-S, Profile of Mood States (POMS)-A, and EI was assessed by Mayer–Salovey–Caruso Emotional Intelligence Test (MSCEIT). At Time 2, negative emotions were induced by nine pictures taken from the International Affective Picture System and participants were asked to complete a second STAI-S and POMS-B questionnaires. At Time 3 participants were allowed to rest doing a distracting task and participants were asked to complete a third STAI-S and POMS-A questionnaires. Results showed that the branches of the MSCEIT emotional facilitation and emotional understanding are related to previous mood states and mood recovery, but not to mood reactivity. This finding contrasts nicely with studies on which emotional recovery was assessed in relation to EI self-reported measures, highlighting the perception and emotional regulation. PMID:26150794

  8. Profiles of Impaired, Spared, and Recovered Neuropsychological Processes in Alcoholism

    Science.gov (United States)

    Oscar-Berman, Marlene; Valmas, Mary M.; Sawyer, Kayle S.; Ruiz, Susan Mosher; Luhar, Riya B.; Gravitz, Zoe R.

    2015-01-01

    Long-term chronic alcoholism is associated with disparate and widespread residual consequences for brain functioning and behavior, and alcoholics suffer a variety of cognitive deficiencies and emotional abnormalities. Alcoholism has heterogeneous origins and outcomes, depending upon factors such as family history, age, gender, and mental or physical health. Consequently, the neuropsychological profiles associated with alcoholism are not uniform among individuals. Moreover, within and across research studies, variability among participants is substantial and contributes to characteristics associated with differential treatment outcomes after detoxification. In order to refine our understanding of alcoholism-related impaired, spared, and recovered abilities, we focus on five specific functional domains: (1) memory, (2) executive functions, (3) emotion and psychosocial skills, (4) visuospatial cognition, and (5) psychomotor abilities. The brain systems that are most vulnerable to alcoholism are the frontocerebellar and mesocorticolimbic circuitries. Over time, with abstinence from alcohol, the brain appears to become reorganized to provide compensation for structural and behavioral deficits. By relying on a combination of clinical and scientific approaches, future research will help to refine the compensatory roles of healthy brain systems, the degree to which abstinence and treatment facilitate the reversal of brain atrophy and dysfunction, and the importance of individual differences to outcome. PMID:25307576

  9. Profiles of impaired, spared, and recovered neuropsychologic processes in alcoholism.

    Science.gov (United States)

    Oscar-Berman, Marlene; Valmas, Mary M; Sawyer, Kayle S; Ruiz, Susan Mosher; Luhar, Riya B; Gravitz, Zoe R

    2014-01-01

    Long-term chronic alcoholism is associated with disparate and widespread residual consequences for brain functioning and behavior, and alcoholics suffer a variety of cognitive deficiencies and emotional abnormalities. Alcoholism has heterogeneous origins and outcomes, depending upon factors such as family history, age, gender, and mental or physical health. Consequently, the neuropsychologic profiles associated with alcoholism are not uniform among individuals. Moreover, within and across research studies, variability among subjects is substantial and contributes to characteristics associated with differential treatment outcomes after detoxification. In order to refine our understanding of alcoholism-related impaired, spared, and recovered abilities, we focus on five specific functional domains: (1) memory; (2) executive functions; (3) emotion and psychosocial skills; (4) visuospatial cognition; and (5) psychomotor abilities. Although the entire brain might be vulnerable in uncomplicated alcoholism, the brain systems that are considered to be most at risk are the frontocerebellar and mesocorticolimbic circuitries. Over time, with abstinence from alcohol, the brain appears to become reorganized to provide compensation for structural and behavioral deficits. By relying on a combination of clinical and scientific approaches, future research will help to refine the compensatory roles of healthy brain systems, the degree to which abstinence and treatment facilitate the reversal of brain atrophy and dysfunction, and the importance of individual differences to outcome. © 2014 Elsevier B.V. All rights reserved.

  10. Can Phosphate Salts Recovered from Manure Replace Conventional Phosphate Fertilizer?

    Directory of Open Access Journals (Sweden)

    Andrea Ehmann

    2017-01-01

    Full Text Available Pig farming produces more manure than can reasonably be spread onto surrounding fields, particularly in regions with high livestock densities and limited land availability. Nutrient recycling offers an attractive solution for dealing with manure excesses and is one main objective of the European commission-funded project “BioEcoSIM”. Phosphate salts (“P-Salt” were recovered from the separated liquid manure fraction. The solid fraction was dried and carbonized to biochar. This study compared the fertilizing performance of P-Salt and conventional phosphate fertilizer and determined whether additional biochar application further increased biomass yields. The fertilizers and biochar were tested in pot experiments with spring barley and faba beans using two nutrient-poor soils. The crops were fertilized with P-Salt at three levels and biochar in two concentrations. Biomass yield was determined after six weeks. Plant and soil samples were analysed for nitrogen, phosphorus and potassium contents. The P-Salt had similar or even better effects than mineral fertilizer on growth in both crops and soils. Slow release of nutrients can prevent leaching, rendering P-Salt a particularly suitable fertilizer for light sandy soils. Biochar can enhance its fertilizing effect, but the underlying mechanisms need further investigation. These novel products are concluded to be promising candidates for efficient fertilization strategies.

  11. Challenges in recovering resources from acid mine drainage

    Science.gov (United States)

    Nordstrom, D. Kirk; Bowell, Robert J.; Campbell, Kate M.; Alpers, Charles N.

    2017-01-01

    Metal recovery from mine waters and effluents is not a new approach but one that has occurred largely opportunistically over the last four millennia. Due to the need for low-cost resources and increasingly stringent environmental conditions, mine waters are being considered in a fresh light with a designed, deliberate approach to resource recovery often as part of a larger water treatment evaluation. Mine water chemistry is highly dependent on many factors including geology, ore deposit composition and mineralogy, mining methods, climate, site hydrology, and others. Mine waters are typically Ca-Mg-SO4±Al±Fe with a broad range in pH and metal content. The main issue in recovering components of these waters having potential economic value, such as base metals or rare earth elements, is the separation of these from more reactive metals such as Fe and Al. Broad categories of methods for separating and extracting substances from acidic mine drainage are chemical and biological. Chemical methods include solution, physicochemical, and electrochemical technologies. Advances in membrane techniques such as reverse osmosis have been substantial and the technique is both physical and chemical. Biological methods may be further divided into microbiological and macrobiological, but only the former is considered here as a recovery method, as the latter is typically used as a passive form of water treatment.

  12. Experimental QR code optical encryption: noise-free data recovering.

    Science.gov (United States)

    Barrera, John Fredy; Mira-Agudelo, Alejandro; Torroba, Roberto

    2014-05-15

    We report, to our knowledge for the first time, the experimental implementation of a quick response (QR) code as a "container" in an optical encryption system. A joint transform correlator architecture in an interferometric configuration is chosen as the experimental scheme. As the implementation is not possible in a single step, a multiplexing procedure to encrypt the QR code of the original information is applied. Once the QR code is correctly decrypted, the speckle noise present in the recovered QR code is eliminated by a simple digital procedure. Finally, the original information is retrieved completely free of any kind of degradation after reading the QR code. Additionally, we propose and implement a new protocol in which the reception of the encrypted QR code and its decryption, the digital block processing, and the reading of the decrypted QR code are performed employing only one device (smartphone, tablet, or computer). The overall method probes to produce an outcome far more attractive to make the adoption of the technique a plausible option. Experimental results are presented to demonstrate the practicality of the proposed security system.

  13. Recovering ethics after 'technics': developing critical text on technology.

    Science.gov (United States)

    Marck, P B

    2000-01-01

    Much modern science and ethics debate is on high-profile problems such as animal organ transplantation, genetic engineering and fetal tissue research, in discourse that assumes technical tones. Other work, such as narrative ethics, expresses the failed promise of technology in the vivid detail of human experience. However, the essential nature of contemporary technology remains largely opaque to our present ethical lens on health care and on society. The limited controversies of modern science and ethics perpetuate 'technics', a technical, problem-solving mindset that fails to grapple successfully with the complexity of technology. A critical dialectic between practice and scholarship widens the ethical conversation in nursing to consider technology as an ongoing set of daily and fundamental moral choices on how we live. Critical text on technology recovers ethics from the limits of technics, and assists nurses to develop an inherent knowledge of technology that is needed to provide ethical care in a technological world. There are overlooked ethical challenges in the mundane, everyday routine activities of professional practice, and these have gone largely unexamined. Ethical behavior is not the display of one's moral rectitude in times of crisis. It is the day-to-day expression of one's commitment to other persons and the ways in which human beings relate to one another in their daily interactions.

  14. [Masticatory system and maxillofacial prosthesis: From pathology to function recovering].

    Science.gov (United States)

    Vo Quang, S; Dichamp, J

    2016-12-01

    Maxillofacial prosthesis (MFP) can be defined as the art and science of esthetic and functional reconstruction of the facial bones, art because it uses hand-crafted and empirical rules, science because of its technical rigorism and its integration in medicine. MFP aims to multidisciplinary rehabilitate patients presenting with cutaneous and underlying structures defects, It also allows for functional speech and swallowing rehabilitation related to temporo-mandibular joint disorders. Whatever the origin, (traumatic, infectious ortumoral), surgical treatment of these TMJ disorders is usually not indicated in first-line. Functional treatment is often sufficient if started early in an observant patient. The aim of our article was to present the different types devices available for the rehabilitation of the masticatory system according to pathology. The first part will treat about the preservation of the TMJ range of motion in a preventive way. A second part will treat about the possibilities to recover the range of motion in a curative way. A third part will treat about mandibular reposition. At last, we will focus on the devices allowing for mandibular kinetic rehabilitation in adults and in a special pediatric case. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  15. Emotional intelligence and recovering from induced negative emotional state

    Directory of Open Access Journals (Sweden)

    Joaquín T. Limonero

    2015-06-01

    Full Text Available The aim of the present study was to examine the relationship between emotional intelligence and recovering from negative emotions induction, using a performance test to measure Emotional Inteligence (EI. Sixty seven undergraduates participated in the procedure, which lasted 75 minutes and was divided into three stages. At Time 1, subjects answered the STAI-S, POMS-A, and EI was assessed by MSCEIT. At Time 2, negative emotions were induced by 9 pictures taken from the International Affective Picture System (IAPS and participants were asked to complete a second STAI-S and POMS-B questionnaires. At Time 3 participants were allowed to rest doing a distracting task and participants were asked to complete a third STAI-S and POMS-A questionnaires. Results showed that the branches of the MSCEIT emotional facilitation and emotional understanding are related to previous mood states and mood recovery, but not to mood reactivity. This finding contrasts nicely with studies on which emotional recovery was assessed in relation to EI self-reported measures, highlighting the perception and emotional regulation.

  16. Bioprocess integration for human mesenchymal stem cells: From up to downstream processing scale-up to cell proteome characterization.

    Science.gov (United States)

    Cunha, Bárbara; Aguiar, Tiago; Carvalho, Sofia B; Silva, Marta M; Gomes, Ricardo A; Carrondo, Manuel J T; Gomes-Alves, Patrícia; Peixoto, Cristina; Serra, Margarida; Alves, Paula M

    2017-04-20

    To deliver the required cell numbers and doses to therapy, scaling-up production and purification processes (at least to the liter-scale) while maintaining cells' characteristics is compulsory. Therefore, the aim of this work was to prove scalability of an integrated streamlined bioprocess compatible with current good manufacturing practices (cGMP) comprised by cell expansion, harvesting and volume reduction unit operations using human mesenchymal stem cells (hMSC) isolated from bone marrow (BM-MSC) and adipose tissue (AT-MSC). BM-MSC and AT-MSC expansion and harvesting steps were scaled-up from spinner flasks to 2L scale stirred tank single-use bioreactor using synthetic microcarriers and xeno-free medium, ensuring high cellular volumetric productivities (50×106cellL-1day-1), expansion factors (14-16 fold) and cell recovery yields (80%). For the concentration step, flat sheet cassettes (FSC) and hollow fiber cartridges (HF) were compared showing a fairly linear scale-up, with a need to slightly decrease the permeate flux (30-50 LMH, respectively) to maximize cell recovery yield. Nonetheless, FSC allowed to recover 18% more cells after a volume reduction factor of 50. Overall, at the end of the entire bioprocess more than 65% of viable (>95%) hMSC could be recovered without compromising cell's critical quality attributes (CQA) of viability, identity and differentiation potential. Alongside the standard quality assays, a proteomics workflow based on mass spectrometry tools was established to characterize the impact of processing on hMSC's CQA; These analytical tools constitute a powerful tool to be used in process design and development. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Recovering stimulus locations using populations of eye-position modulated neurons in dorsal and ventral visual streams of nonhuman primates

    Directory of Open Access Journals (Sweden)

    Anne B Sereno

    2014-03-01

    Full Text Available We recorded visual responses while monkeys fixated the same target at different gaze angles, both dorsally (lateral intraparietal cortex, LIP and ventrally (anterior inferotemporal cortex, AIT. While eye-position modulations occurred in both areas, they were both more frequent and stronger in LIP neurons. We used an intrinsic population decoding technique, multidimensional scaling, to recover eye positions, equivalent to recovering fixated target locations. We report that eye-position based visual space in LIP was more accurate (i.e., metric. Nevertheless, the AIT spatial representation remained largely topologically correct, perhaps indicative of a categorical spatial representation (i.e., a qualitative description such as left of or above as opposed to a quantitative, metrically precise description. Additionally, we developed a simple neural model of eye position signals and illustrate that differences in single cell characteristics can influence the ability to recover target position in a population of cells. We demonstrate for the first time that the ventral stream contains sufficient information for constructing an eye-position based spatial representation. Furthermore we demonstrate, in dorsal and ventral streams as well as modeling, that target locations can be extracted directly from eye position signals in cortical visual responses without computing coordinate transforms of visual space.

  18. Aging after noise exposure: acceleration of cochlear synaptopathy in "recovered" ears.

    Science.gov (United States)

    Fernandez, Katharine A; Jeffers, Penelope W C; Lall, Kumud; Liberman, M Charles; Kujawa, Sharon G

    2015-05-13

    Cochlear synaptic loss, rather than hair cell death, is the earliest sign of damage in both noise- and age-related hearing impairment (Kujawa and Liberman, 2009; Sergeyenko et al., 2013). Here, we compare cochlear aging after two types of noise exposure: one producing permanent synaptic damage without hair cell loss and another producing neither synaptopathy nor hair cell loss. Adult mice were exposed (8-16 kHz, 100 or 91 dB SPL for 2 h) and then evaluated from 1 h to ∼ 20 months after exposure. Cochlear function was assessed via distortion product otoacoustic emissions and auditory brainstem responses (ABRs). Cochlear whole mounts and plastic sections were studied to quantify hair cells, cochlear neurons, and the synapses connecting them. The synaptopathic noise (100 dB) caused 35-50 dB threshold shifts at 24 h. By 2 weeks, thresholds had recovered, but synaptic counts and ABR amplitudes at high frequencies were reduced by up to ∼ 45%. As exposed animals aged, synaptopathy was exacerbated compared with controls and spread to lower frequencies. Proportional ganglion cell losses followed. Threshold shifts first appeared >1 year after exposure and, by ∼ 20 months, were up to 18 dB greater in the synaptopathic noise group. Outer hair cell losses were exacerbated in the same time frame (∼ 10% at 32 kHz). In contrast, the 91 dB exposure, producing transient threshold shift without acute synaptopathy, showed no acceleration of synaptic loss or cochlear dysfunction as animals aged, at least to ∼ 1 year after exposure. Therefore, interactions between noise and aging may require an acute synaptopathy, but a single synaptopathic exposure can accelerate cochlear aging. Copyright © 2015 the authors 0270-6474/15/357509-12$15.00/0.

  19. Resource recovery and reuse as an incentive for a more viable sanitation service chain

    Directory of Open Access Journals (Sweden)

    Krishna C. Rao

    2017-06-01

    Full Text Available Recovering nutrients, water and energy from domestic waste streams, including wastewater and faecal sludge, is slowly gaining momentum in low-income countries. Resource recovery and reuse (RRR offers value beyond environmental benefits through cost recovery. An expected game changer in sanitation service provision is a business model where benefits accrued via RRR can support upstream sanitation services despite the multitude of private and public stakeholders involved from waste collection to treatment. This paper shows options of how resource recovery and reuse can be an incentive for the sustainable sanitation service chain, by recovering costs where revenue can feed back internally or using generated revenues from reuse to fill financial gaps across the service chain to complement other supporting mechanisms for making waste management more attractive.

  20. Resource recovery and reuse as an incentive for a more viable sanitation service chain

    OpenAIRE

    Krishna C. Rao; Miriam Otoo; Pay Drechsel; Hanjra, Munir A.

    2017-01-01

    Recovering nutrients, water and energy from domestic waste streams, including wastewater and faecal sludge, is slowly gaining momentum in low-income countries. Resource recovery and reuse (RRR) offers value beyond environmental benefits through cost recovery. An expected game changer in sanitation service provision is a business model where benefits accrued via RRR can support upstream sanitation services despite the multitude of private and public stakeholders involved from waste...

  1. [Structural recovering in Andean successional forests from Porce (Antioquia, Colombia)].

    Science.gov (United States)

    Yepes, Adriana P; del Valle, Jorge I; Jaramillo, Sandra L; Orrego, Sergio A

    2010-03-01

    Places subjected to natural or human disturbance can recover forest through an ecological process called secondary succession. Tropical succession is affected by factors such as disturbances, distance from original forest, surface configuration and local climate. These factors determine the composition of species and the time trend of the succession itself. We studied succession in soils used for cattle ranching over various decades in the Porce Region of Colombia (Andean Colombian forests). A set of twenty five permanent plots was measured, including nine plots (20 x 50 m) in primary forests and sixteen (20 x 25 m) in secondary forests. All trees with diameter > or =1.0 cm were measured. We analyzed stem density, basal area, above-ground biomass and species richness, in a successional process of ca. 43 years, and in primary forests. The secondary forests' age was estimated in previous studies, using radiocarbon dating, aerial photographs and a high-resolution satellite image analysis (7 to >43 years). In total, 1,143 and 1,766 stems were measured in primary and secondary forests, respectively. Basal area (5.7 to 85.4 m2 ha(-1)), above-ground biomass (19.1 to 1,011.5 t ha(-1)) and species richness (4 to 69) directly increased with site age, while steam density decreased (3,180 to 590). Diametric distributions were "J-inverted" for primary forests and even-aged size-class structures for secondary forests. Three species of palms were abundant and exclusive in old secondary forests and primary forests: Oenocarpus mapora, Euterpe precatoria and Oenocarpus bataua. These palms happened in cohorts after forest disturbances. Secondary forest structure was 40% in more than 43 years of forest succession and indicate that many factors are interacting and affecting the forests succession in the area (e.g. agriculture, cattle ranching, mining, etc.).

  2. Persistence and potential Viable but Non-culturable state of pathogenic bacteria during storage of digestates from agricultural biogas plants

    Directory of Open Access Journals (Sweden)

    Geraldine Maynaud

    2016-09-01

    Full Text Available Despite the development of on-farm anaerobic digestion as a process for making profitable use of animal by-products, factors leading to the inactivation of pathogenic bacteria during storage of digestates remain poorly described. Here, a microcosm approach was used to evaluate the persistence of three pathogenic bacteria (Salmonella enterica Derby, Campylobacter coli and Listeria monocytogenes in digestates from farms, stored for later land spreading. Nine samples, including raw digestates, liquid fractions of digestate and composted digestates, were inoculated with each pathogen and maintained for 40 days at 24°C. Concentrations of pathogens were monitored using culture and qPCR methods. The persistence of L. monocytogenes, detected up to 20 days after inoculation, was higher than that of Salmonella Derby, detected for 7-20 days, and of C. coli (not detected after 7 days. In some digestates, the concentration of the pathogens by qPCR assay was several orders of magnitude higher than the concentration of culturable cells, suggesting a potential loss of culturability and induction of Viable but Non-Culturable (VBNC state. The potential VBNC state which was generally not observed in the same digestate for the three pathogens, occurred more frequently for C. coli and L. monocytogenes than for Salmonella Derby. Composting a digestate reduced the persistence of seeded L. monocytogenes but promoted the maintenance of Salmonella Derby. The effect of NH4+/NH3 on the culturability of C. coli and Salmonella Derby was also shown.The loss of culturability may be the underlying mechanism for the regrowth of pathogens. We have also demonstrated the importance of using molecular tools to monitor pathogens in environmental samples since culture methods may underestimate cell concentration. Our results underline the importance of considering VBNC cells when evaluating the sanitary effect of an anaerobic digestion process and the persistence of pathogens during

  3. An Extension of the Hawkins and Simon Condition Characterizing Viable Techniques

    Directory of Open Access Journals (Sweden)

    Alberto Benítez Sánchez

    2015-01-01

    Full Text Available This paper discusses an extended version of the Hawkins and Simon condition which constitutes a synthetic formulation of the mathematical properties that viable economies must satisfy in single production models. The new version is implicit in the economic interpretations offered by them of the Hawkins and Simon condition, once a correction is introduced in one of those interpretations. Moreover, the paper details the meaning of the extended version following the interpretation of the original version proposed by Dorfman, Samuelson, and Solow. It also introduces a characteristic property of indecomposable matrices that has not previously been published.

  4. THE CIVILIZATIONAL PROJECT AND lTS DISCONTENTS: TOWARD A VIABLE GLOBAL MARKET SOCIETY?

    Directory of Open Access Journals (Sweden)

    Volker Bornschier

    2015-08-01

    Full Text Available I agree with the point of an anonymous reviewer arguing that a pro-found discussion on the prerequisites of a viable global market society has too many facets to be contained within the bounds of a single journal article. Yet, in order to enter the debate now we should not wait until book length treatments become available-often only after years. This is, therfore, an essay attempting to overview various conflicts and contradictions within the global social system. It sythesizes arguments developed in more detail else-where.

  5. Fake Journals: Their Features and Some Viable Ways to Distinguishing Them

    DEFF Research Database (Denmark)

    Hemmat Esfe, Mohammad; Wongwises, Somchai; Asadi, Amin

    2015-01-01

    In this paper, we aim to discuss the fake journals and their advertisement and publication techniques. These types of journals mostly start and continue their activities by using the name of some indexed journals and establishing fake websites. The fake journals and publishers, while asking...... the authors for a significant amount of money for publishing their papers, have no peer-review process, publish the papers without any revision on the fake sites, and put the scientific reputation and prestige of the researchers in jeopardy. In the rest of the paper, we present some viable techniques in order...

  6. Information Warfare: using the viable system model as a framework to attack organisations

    Directory of Open Access Journals (Sweden)

    Bill Hutchinson

    2002-05-01

    Full Text Available Information is the glue in any organization. It is needed for policy, decision-making, control, and co-ordination. If an organisation's information systems are disrupted or destroyed, then damage to the whole inevitably follows. This paper uses a proven systemic, analytic framework the Viable System Model (VSM - in a functionalist mode, to analyse the vulnerabilities of an organisation's information resources to this form of aggression. It examines the tactics available, and where they can be used to effectively attack an organisation.

  7. Current progress and challenges in engineering viable artificial leaf for solar water splitting

    Directory of Open Access Journals (Sweden)

    Phuc D. Nguyen

    2017-12-01

    Full Text Available Large scale production of H2, a clean fuel, can be realized with just water and solar light energy by employing a viable energy conversion device called artificial leaf. In this tutorial review, we discuss on advances achieved recently and technical challenges remained toward the creation of such a leaf. Development of key components like catalysts for water electrolysis process and light harvester for harvesting solar energy as well as strategies being developed for assembling these components to create a complete artificial leaf will be highlighted.

  8. Emergency total thyroidectomy for bleeding anaplastic thyroid carcinoma: A viable option for palliation

    Directory of Open Access Journals (Sweden)

    Sunil Kumar

    2011-01-01

    Full Text Available Anaplastic thyroid carcinoma (ATC is a rare and highly aggressive thyroid neoplasm. Bleeding from tumor is an uncommon, but potentially life-threatening complication requiring sophisticated intervention facilities which are not usually available at odd hours in emergency. We report the case of a 45-year-old woman who presented with exsanguinating hemorrhage from ATC and was treated by emergency total thyroidectomy. The patient is well three months postoperatively. Emergency total thyroidectomy is a viable option for palliation in ATC presenting with bleeding.

  9. Texture analysis of cardiac cine magnetic resonance imaging to detect non-viable segments in patients with chronic myocardial infarction.

    Science.gov (United States)

    Larroza, Andrés; López-Lereu, María P; Monmeneu, José V; Gavara, Jose; Chorro, Francisco J; Bodí, Vicente; Moratal, David

    2018-02-01

    To investigate the ability of texture analysis to differentiate between infarcted non-viable, viable, and remote segments on cardiac cine magnetic resonance imaging (MRI). This retrospective study included 50 patients suffering chronic myocardial infarction. The data was randomly split into training (30 patients) and testing (20 patients) sets. The left ventricular myocardium was segmented according to the 17-segment model in both cine and late gadolinium enhancement (LGE) MRI. Infarcted myocardium regions were identified on LGE in short-axis views. Non-viable segments were identified as those showing LGE ≥ 50%, and viable segments those showing 0 cine images. A support vector machine (SVM) classifier was trained with different combination of texture features to obtain a model that provided optimal classification performance. The best classification on testing set was achieved with local binary patterns features using a 2D + t approach, in which the features are computed by including information of the time dimension available in cine sequences. The best overall area under the receiver operating characteristic curve (AUC) were: 0.849, sensitivity of 92% to detect non-viable segments, 72% to detect viable segments, and 85% to detect remote segments. Non-viable segments can be detected on cine MRI using texture analysis and this may be used as hypothesis for future research aiming to detect the infarcted myocardium by means of a gadolinium-free approach. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  10. Reduction of viable Enterococcus faecalis in human radicular dentin treated with 1% cetrimide and conventional intracanal medicaments.

    Science.gov (United States)

    Carbajal Mejía, Jeison B; Aguilar Arrieta, Angela

    2016-08-01

    The purpose of this study was to evaluate the viability of E. faecalis after a 14-day exposure to 1% cetrimide (CET), triantibiotic paste (i.e., metronidazole, minocycline, and ciprofloxacin), 2% chlorhexidine (CHX) gel, and calcium hydroxide (Ca[OH]2 ) in an infected dentine model. A total of 75 roots of extracted uniradicular human teeth were chemomechanically prepared, sterilized, and infected for 21 days with E. faecalis. Samples were divided into five groups (n = 15) to apply intracanal medicaments, namely saline solution (negative control), 1% CET, triantibiotic paste (TRIA), 2% CHX gel, and Ca(OH)2 during 14 days. Dentine samples were collected and stained with the SYTO 9/propidium iodide technique (Live/Dead, Bacligth, Invitrogen, Eugene, OR, USA) for fluorescence microscopy to obtain the percentage of viable cells. Statistical analysis was performed using one-way analysis of variance followed by Tukey's multiple comparison test (P faecalis when compared with the control group. In addition, the least cytotoxic medicament was Ca(OH)2 followed by CHX. There was no significant difference between 1% CET and TRIA (P = 0.98). Both 1% CET and TRIA significantly reduced the viability of E. faecalis in dentine of extracted teeth in comparison with 2% CHX gel and calcium hydroxide paste. Further laboratory and clinical investigations should be carried out to validate findings of the beneficial use of 1% CET as an intracanal medicament against E. faecalis. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  11. Isolation of Viable but Non-culturable Bacteria from Printing and Dyeing Wastewater Bioreactor Based on Resuscitation Promoting Factor.

    Science.gov (United States)

    Jin, Yi; Gan, Guojuan; Yu, Xiaoyun; Wu, Dongdong; Zhang, Li; Yang, Na; Hu, Jiadan; Liu, Zhiheng; Zhang, Lixin; Hong, Huachang; Yan, Xiaoqing; Liang, Yan; Ding, Linxian; Pan, Yonglong

    2017-07-01

    Printing and dyeing wastewater with high content of organic matters, high colority, and poor biochemical performance is hard to be degraded. In this study, we isolated viable but non-culturable (VBNC) bacteria from printing and dyeing wastewater with the culture media contained resuscitation promoting factor (Rpf) protein secreted by Micrococcus luteus, counted the culturable cells number with the most probable number, sequenced 16S rRNA genes, and performed polymerase chain reaction-denaturing gradient gel electrophoresis. It is obviously that the addition of Rpf in the enrichment culture could promote growth and resuscitation of bacteria in VBNC state to obtain more fastidious bacteria significantly. The identified bacteria were assigned to nine genera in the treatment group, while the two strains of Ochrobactrum anthropi and Microbacterium sp. could not be isolated from the control group. The function of isolated strains was explored and these strains could degrade the dye of Congo red. This study provides a new sight into the further study including the present state, composition, formation mechanism, and recovery mechanism about VBNC bacteria in printing and dyeing wastewater, which would promote to understand bacterial community in printing and dyeing wastewater, and to obtain VBNC bacteria from ecological environment.

  12. Are big potassium-type Ca(2+)-activated potassium channels a viable target for the treatment of epilepsy?

    Science.gov (United States)

    Leo, Antonio; Citraro, Rita; Constanti, Andrew; De Sarro, Giovambattista; Russo, Emilio

    2015-07-01

    BK (big potassium) channels are Ca(2+)-activated K(+) channels widely expressed in mammalian cells. They are extensively distributed in the CNS, the most abundant level being found in brain areas largely involved in epilepsy, namely cortex, hippocampus, piriform cortex, and other limbic structures. BK channels control action potential shape/duration, thereby regulating membrane excitability and Ca(2+) signaling. The potassium channel superfamily represents a rich source of potential targets for therapeutic intervention in epilepsy. Some studies have identified alterations in BK channel function, therefore, supporting the development of drugs acting on these channels for epilepsy treatment. The actual sketch is intriguing and controversial, since mechanisms altering the physiological role of BK channels leading to either a loss- or gain-of-function have both been linked to seizure onset. Not many studies have been performed to unravel the efficacy of drugs acting on these channels as potential antiepileptics; however, paradoxically, efficacy has been demonstrated for both BK channel openers and blockers. Furthermore, their potential usefulness in preventing epileptogenesis has not been investigated at all. Substantial data on risks and benefits of modulating these channels are urgently needed to draw a definitive conclusion on whether BK channels are a viable future target for the treatment of epilepsy.

  13. Structure based modification of Bluetongue virus helicase protein VP6 to produce a viable VP6-truncated BTV

    Energy Technology Data Exchange (ETDEWEB)

    Matsuo, Eiko [Microbiology and Immunology, Division of Animal Science, Department of Bioresource Science, Graduate School of Agricultural Science, Kobe University, 1-1, Rokkodai, Nada-ku, Kobe-City 657-8501 (Japan); Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT (United Kingdom); Leon, Esther; Matthews, Steve J. [Division of Molecular Biosciences, Centre for Structural Biology, Imperial College London, South Kensington, London SW7 2AZ (United Kingdom); Roy, Polly, E-mail: polly.roy@lshtm.ac.uk [Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT (United Kingdom)

    2014-09-05

    Highlights: • NMR analysis on BTV VP6 reveals two large loop regions. • The loss of a loop (aa 34–130) does not affect the overall fold of the protein. • A region of VP6 (aa 34–92) is not required for BTV replication. • A region of VP6 (aa 93–130) plays an essential role in the virus replication. - Abstract: Bluetongue virus core protein VP6 is an ATP hydrolysis dependent RNA helicase. However, despite much study, the precise role of VP6 within the viral capsid and its structure remain unclear. To investigate the requirement of VP6 in BTV replication, we initiated a structural and biological study. Multinuclear nuclear magnetic resonance spectra were assigned on his-tagged full-length VP6 (329 amino acid residues) as well as several truncated VP6 variants. The analysis revealed a large structured domain with two large loop regions that exhibit significant conformational exchange. One of the loops (amino acid position 34–130) could be removed without affecting the overall fold of the protein. Moreover, using a BTV reverse genetics system, it was possible to demonstrate that the VP6-truncated BTV was viable in BHK cells in the absence of any helper VP6 protein, suggesting that a large portion of this loop region is not absolutely required for BTV replication.

  14. Bacteriophage treatment significantly reduces viable Clostridium difficile and prevents toxin production in an in vitro model system.

    Science.gov (United States)

    Meader, Emma; Mayer, Melinda J; Gasson, Michael J; Steverding, Dietmar; Carding, Simon R; Narbad, Arjan

    2010-12-01

    Clostridium difficile is primarily a nosocomial pathogen, causing thousands of cases of antibiotic-associated diarrhoea in the UK each year. In this study, we used a batch fermentation model of a C. difficile colonised system to evaluate the potential of a prophylactic and a remedial bacteriophage treatment regime to control the pathogen. It is shown that the prophylaxis regime was effective at preventing the growth of C. difficile (p = <0.001) and precluded the production of detectable levels of toxins A and B. The remedial treatment regime caused a less profound and somewhat transient decrease in the number of viable C. difficile cells (p = <0.0001), but still resulted in a lower level of toxin production relative to the control. The numbers of commensal bacteria including total aerobes and anaerobes, Bifidobacterium sp., Bacteroides sp., Lactobacillus sp., total Clostridium sp., and Enterobacteriaceae were not significantly decreased by this therapy, whereas significant detrimental effects were observed with metronidazole treatment. Our study indicates that phage therapy has potential to be used for the control of C. difficile; it highlights the main benefits of this approach, and some future challenges. Copyright © 2010 Elsevier Ltd. All rights reserved.

  15. Stem Cell Transplants (For Teens)

    Science.gov (United States)

    ... Situations Talking to Your Parents - or Other Adults Stem Cell Transplants KidsHealth > For Teens > Stem Cell Transplants Print ... Does it Take to Recover? Coping What Are Stem Cells? As you probably remember from biology class, every ...

  16. Recovering Drug-Induced Apoptosis Subnetwork from Connectivity Map Data

    Directory of Open Access Journals (Sweden)

    Jiyang Yu

    2015-01-01

    Full Text Available The Connectivity Map (CMAP project profiled human cancer cell lines exposed to a library of anticancer compounds with the goal of connecting cancer with underlying genes and potential treatments. Since the therapeutic goal of most anticancer drugs is to induce tumor-selective apoptosis, it is critical to understand the specific cell death pathways triggered by drugs. This can help to better understand the mechanism of how cancer cells respond to chemical stimulations and improve the treatment of human tumors. In this study, using CMAP microarray data from breast cancer cell line MCF7, we applied a Gaussian Bayesian network modeling approach and identified apoptosis as a major drug-induced cellular-pathway. We then focused on 13 apoptotic genes that showed significant differential expression across all drug-perturbed samples to reconstruct the apoptosis network. In our predicted subnetwork, 9 out of 15 high-confidence interactions were validated in the literature, and our inferred network captured two major cell death pathways by identifying BCL2L11 and PMAIP1 as key interacting players for the intrinsic apoptosis pathway and TAXBP1 and TNFAIP3 for the extrinsic apoptosis pathway. Our inferred apoptosis network also suggested the role of BCL2L11 and TNFAIP3 as “gateway” genes in the drug-induced intrinsic and extrinsic apoptosis pathways.

  17. Viable Reserve Networks Arise From Individual Landholder Responses To Conservation Incentives

    Directory of Open Access Journals (Sweden)

    Kenneth M. Chomitz

    2006-12-01

    Full Text Available Conservation in densely settled biodiversity hotspots often requires setting up reserve networks that maintain sufficient contiguous habitat to support viable species populations. Because it is difficult to secure landholder compliance with a tightly constrained reserve network design, attention has shifted to voluntary incentive mechanisms, such as purchase of conservation easements by reverse auction or through a fixed-price offer. These mechanisms carry potential advantages of transparency, simplicity, and low cost. However, uncoordinated individual response to these incentives has been assumed incompatible with the conservation goal of viability, which depends on contiguous habitat and biodiversity representation. We model such incentives for southern Bahia in the Brazilian Atlantic Forest, one of the biologically richest and most threatened global biodiversity hotspots. Here, forest cover is spatially autocorrelated and associated with depressed land values, a situation that may be characteristic of long-settled areas with forests fragmented by agriculture. We find that in this situation, a voluntary incentive system can yield a reserve network characterized by large, viable patches of contiguous forest, and representation of subregions with distinct vegetation types and biotic assemblages, without explicit planning for those outcomes.

  18. Generation of viable progeny from dead brooders of endangered catfish Clarias magur (Hamilton, 1822

    Directory of Open Access Journals (Sweden)

    Sullip Kumar Majhi

    2017-10-01

    Full Text Available The obligatory air-breathing catfish Clarias magur is a prime candidate for aquaculture owing to its unique taste, high growth rate, and hardy nature. However, recently the IUCN has listed the species under the endangered category because the population has critically declined in the wild. The sexually mature C. magur brooders are often collected from their natural habitats for seed production in captivity. In many cases, the brooder dies due to handling injuries or confinement stress. In this study, we demonstrated that viable progeny could be generated from freshly dead sexually mature C. magur. Three hours after death, the gonads were excised, macroscopically examined and gamete viability was evaluated. Artificial fertilization was performed by mixing the sperm suspension with the eggs. Water was added after 1 min of mixing to activate the fertilization process. We observed 85%-93% fertilization success from gametes derived from dead donors as opposed to 90%-95% from those derived from live control donors. The embryos showed normal development and resulted in the generation of 88%-92% viable progeny, which was similar to the progeny derived from control donors (92%-93%. The results obtained in this study will have profound implications in enhancing the seed production of endangered C. magur and could potentially be applied to other key commercially or endangered fish species. Keywords: Biological sciences, Developmental biology, Zoology

  19. Economically Viable Components from Jerusalem Artichoke (Helianthus tuberosus L. in a Biorefinery Concept

    Directory of Open Access Journals (Sweden)

    Eva Johansson

    2015-04-01

    Full Text Available Biorefinery applications are receiving growing interest due to climatic and waste disposal issues and lack of petroleum resources. Jerusalem artichoke (Helianthus tuberosus L. is suitable for biorefinery applications due to high biomass production and limited cultivation requirements. This paper focuses on the potential of Jerusalem artichoke as a biorefinery crop and the most viable products in such a case. The carbohydrates in the tubers were found to have potential for production of platform chemicals, e.g., succinic acid. However, economic analysis showed that production of platform chemicals as a single product was too expensive to be competitive with petrochemically produced sugars. Therefore, production of several products from the same crop is a must. Additional products are protein based ones from tubers and leaves and biogas from residues, although both are of low value and amount. High bioactive activity was found in the young leaves of the crop, and the sesquiterpene lactones are of specific interest, as other compounds from this group have shown inhibitory effects on several human diseases. Thus, future focus should be on understanding the usefulness of small molecules, to develop methods for their extraction and purification and to further develop sustainable and viable methods for the production of platform chemicals.

  20. Economically viable components from Jerusalem artichoke (Helianthus tuberosus L.) in a biorefinery concept.

    Science.gov (United States)

    Johansson, Eva; Prade, Thomas; Angelidaki, Irini; Svensson, Sven-Erik; Newson, William R; Gunnarsson, Ingólfur Bragi; Hovmalm, Helena Persson

    2015-04-22

    Biorefinery applications are receiving growing interest due to climatic and waste disposal issues and lack of petroleum resources. Jerusalem artichoke (Helianthus tuberosus L.) is suitable for biorefinery applications due to high biomass production and limited cultivation requirements. This paper focuses on the potential of Jerusalem artichoke as a biorefinery crop and the most viable products in such a case. The carbohydrates in the tubers were found to have potential for production of platform chemicals, e.g., succinic acid. However, economic analysis showed that production of platform chemicals as a single product was too expensive to be competitive with petrochemically produced sugars. Therefore, production of several products from the same crop is a must. Additional products are protein based ones from tubers and leaves and biogas from residues, although both are of low value and amount. High bioactive activity was found in the young leaves of the crop, and the sesquiterpene lactones are of specific interest, as other compounds from this group have shown inhibitory effects on several human diseases. Thus, future focus should be on understanding the usefulness of small molecules, to develop methods for their extraction and purification and to further develop sustainable and viable methods for the production of platform chemicals.

  1. Recovering the Atmospheric Resources of Mars: Updating the MARRS Study

    Science.gov (United States)

    England, Christopher; Hrubes, J. Dana

    2006-01-01

    In 2000 a conceptual design was conducted of a plant that extracts oxygen (O2) directly from the martian atmosphere, and that makes water and carbon monoxide (CO) as by-products. Updated estimates suggest that the amount of O2 in the atmosphere is about 2.3 times greater than that used as the basis for the 2000 study. In this paper, estimates for O2 and by-products, and for energy and mass requirements based on the higher O2 value are updated. The basis for the design, termed ``MARRS'' for Mars Atmosphere Resource Recovery System, is the NASA/JSC Mars Reference Mission (MRM) requirement for O2, estimated at 5.8 kg/hr for about 500 sols. The 2000 study based its design on an atmospheric O2 content of 0.13%, the then-accepted value. Analysis now places the O2 content at about 0.3%, reducing the amount of energy and equipment proportionately. The revised estimate of the thermal power to meet MRM requirements for O2 is an average of about 52 kW, seasonally variable. The new mass estimate is 7898 kg, down from 13650 kg. The new estimate of oxygen content correspondingly reduces the amounts of by-products that can be recovered. CO, a primary fuel and propellant precursor, is produced at about 0.2 kg/kg O2. Water, possibly available at about 0.04 kg/kg O2, is believed not recoverable by the MARRS process at this lower level, even seasonally. An equation is provided for the seasonal variation in atmospheric O2 fraction based on Viking pressure measurements. Oxygen varies seasonally from about 0.25% or 0.34%, the variability affecting plant design. While the higher O2 fraction means reduced amounts of by-products from the MARRS process, large amounts of nitrogen (liquid and gas), argon gas and liquid carbon dioxide (CO2) remain available as by-products for use as respiratory agents, refrigerants, propellants, propellant precursors and working fluids for emergency or backup power, transportation, and surface operations such as drilling.

  2. Energy Expenditure and Physical Activity in Recovering Malnourished Infants

    Directory of Open Access Journals (Sweden)

    Russell Rising

    2010-01-01

    Full Text Available Background. Malnourished infants are small for age and weight. Objectives. Determine profiles in 24-hour energy metabolism in recovering malnourished infants and compare to similarly aged healthy controls. Methods. 10 malnourished infants (58.1±5.9 cm, 7.7±5.6 months were healthy prior to spending 22 hours in the Enhanced Metabolic Testing Activity Chamber for measurement of EE (kcal/min, sleeping metabolic rate (SMR; kcal/min, respiratory quotient (RQ; VCO2/VO2, and physical activity (PA; oscillations in wt/min/kg body weight. Metabolic data were extrapolated to 24 hours (kcal/kg/d. Energy intake (kcal/kg/d and the proportions (% of carbohydrate, protein, and fat were calculated. Anthropometrics for malnourished infants were obtained. Statistical differences (P<.05 between groups were determined (SPSS, version 13. Results. In comparison to controls, malnourished infants were lighter (4.1±1.2 versus 7.3±0.8 kg; P<.05, had less body fat % (10.3±7.6 versus 25.7±2.5, and lower BMI (12.0±1.7 versus 15.5±1.5; P<.05. In contrast, they had greater energy intake (142.7±14.6 versus 85.1±25.8; P<.05 with a greater percentage of carbohydrates (55.1±3.9 versus 47.2±5.2; P<.05. However, malnourished infants had greater 24-hour EE (101.3±20.1 versus 78.6±8.4; P<.05, SMR (92.6±17.1 versus 65.0±3.9; P<.05, and RQ (1.00±0.13 versus 0.86±0.08; P<.05 along with a lower amount of PA (2.3±0.94 versus 4.0±1.5; P<.05. Conclusions. Malnourished infants require more energy, possibly for growth.

  3. In vivo study of the HC-TN strain of hepatitis C virus recovered from a patient with fulminant hepatitis: RNA transcripts of a molecular clone (pHC-TN) are infectious in chimpanzees but not in Huh7.5 cells

    DEFF Research Database (Denmark)

    Sakai, Akito; Takikawa, Shingo; Thimme, Robert

    2007-01-01

    Both viral and host factors are thought to influence the pathogenesis of hepatitis C virus (HCV) infection. We studied strain HC-TN (genotype 1a), which caused fulminant hepatic failure in a patient and, subsequently, severe hepatitis in a chimpanzee (CH1422), to analyze the relationship between...... disease severity, host immune response, viral evolution, and outcome. A second chimpanzee (CH1581) was infected from CH1422 plasma, and a third chimpanzee (CH1579) was infected from RNA transcripts of a consensus cDNA of HC-TN (pHC-TN). RNA transcripts of pHC-TN did not replicate in Huh7.5 cells, which...

  4. Isolation and Identification of Adenovirus Recovered from the Stool ...

    African Journals Online (AJOL)

    In order to establish the role of adenovirus in gastroenteritis in Nigerian children, stool samples were collected from 138 young children with gastroenteritis and 29 other age-matched controls. The samples were inoculated into 6 different tissue culture cell lines and isolates with characteristic CPE were subjected to CFT ...

  5. Prevalence and course of sexual relationship difficulties in recovered and non-recovered patients with borderline personality disorder over 16 years of prospective follow-up.

    Science.gov (United States)

    Karan, Esen; Niesten, Isabella J M; Frankenburg, Frances R; Fitzmaurice, Garrett M; Zanarini, Mary C

    2016-08-01

    Although borderline patients experience a wide range of sexual problems, including promiscuity, there is less evidence documenting their sexual relationship difficulties. This study had two aims. The first was to examine the prevalence of these difficulties (i.e. avoidance of sex and being symptomatic after sex) over 16 years of prospective follow-up among recovered and non-recovered patients with borderline personality disorder (BPD). The second was to determine time-to-remission, recurrence and new onset of these sexual relationship difficulties. The sexual relationship difficulties of 290 patients meeting both DIB-R and DSM-III-R criteria for BPD were assessed at baseline using the Abuse History Interview and reassessed every two years over eight waves of prospective follow-up. The prevalence of sexual relationship difficulties declined significantly over time for both groups of patients, while remaining significantly more common among non-recovered patients. By 16-year follow-up, over 95% of each group achieved remission for both types of difficulties. Recurrences of avoidance of sex were significantly more common in non-recovered patients. Non-recovered patients had higher rates of new onsets compared to recovered patients for each type of sexual relationship difficulty. Taken together, the results suggest that sexual relationship difficulties are not chronic for those with BPD regardless of recovery status. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  6. Viable offspring after imaging of Ca2+ oscillations and visualization of the cortical reaction in mouse eggs.

    Science.gov (United States)

    Satouh, Yuhkoh; Nozawa, Kaori; Yamagata, Kazuo; Fujimoto, Takao; Ikawa, Masahito

    2017-03-01

    During mammalian fertilization, egg Ca 2+ oscillations are known to play pivotal roles in triggering downstream events such as resumption of the cell cycle and the establishment of blocks to polyspermy. However, viable offspring have not been obtained after monitoring Ca 2+ oscillations, and their spatiotemporal links to subsequent events are still to be examined. Therefore, the development of imaging methods to avoid phototoxic damage while labeling these events is required. Here, we examined the usefulness of genetically encoded Ca 2+ indicators for optical imaging (GECOs), in combination with spinning-disk confocal imaging. The Ca 2+ imaging of fertilized mouse eggs with GEM-, G-, or R-GECO recorded successful oscillations (8.19 ± 0.31, 7.56 ± 0.23, or 7.53 ± 0.27 spikes in the first 2 h, respectively), similar to those obtained with chemical indicators. Then, in vitro viability tests revealed that imaging with G- or R-GECO did not interfere with the rate of development to the blastocyst stage (61.8 or 70.0%, respectively, vs 75.0% in control). Furthermore, two-cell transfer to recipient female mice after imaging with G- or R-GECO resulted in a similar birthrate (53.3 or 52.0%, respectively) to that of controls (48.7%). Next, we assessed the quality of the cortical reaction (CR) in artificially activated or fertilized eggs using fluorescently labeled Lens culinaris agglutinin fluorescein isothiocyanate. Multicolor imaging demonstrated that the first few Ca 2+ spikes are sufficient for the completion of the CR and subsequent hardening of the zona pellucida in mouse eggs. These methods provide a framework for studying Ca 2+ dynamics in mammalian fertilization.

  7. Teaching general problem-solving skills is not a substitute for, or a viable addition to, teaching mathematics

    NARCIS (Netherlands)

    Sweller, John; Clark, Richard; Kirschner, Paul A.

    2010-01-01

    Sweller, J., Clark, R., & Kirschner, P. A. (2010). Teaching general problem-solving skills is not a substitute for, or a viable addition to, teaching mathematics. Notices of the American Mathematical Society, 57, 1303-1304.

  8. The Belem Framework for Action: Harnessing the Power and Potential of Adult Learning and Education for a Viable Future

    Science.gov (United States)

    Adult Learning, 2012

    2012-01-01

    This article presents the Belem Framework for Action. This framework focuses on harnessing the power and potential of adult learning and education for a viable future. This framework begins with a preamble on adult education and towards lifelong learning.

  9. Changes in the relative population size of selected ruminal bacteria following an induced episode of acidosis in beef heifers receiving viable and non-viable active dried yeast.

    Science.gov (United States)

    Mohammed, R; Vyas, D; Yang, W Z; Beauchemin, K A

    2017-06-01

    To characterize the changes in the relative population size (RPS) of select ruminal bacteria and rumen fermentation variables in beef heifers supplemented with a strain of Saccharomyces cerevisiae as viable active dried (ADY) or killed dried (KDY) yeast following an induced episode of ruminal acidosis. Six ruminally cannulated beef heifers fed a diet consisting of 50% forage and 50% grain (dry matter basis) were used in a replicated 3 × 3 Latin square design with three 28-day periods. Treatments were: (i) control (CTRL; no yeast); (ii) ADY (4 g day-1 providing 1010  CFU per g; AB Vista, UK); and (iii) KDY (4 g day-1 autoclaved ADY). The acidosis challenge was induced on day 22 and rumen samples were collected on day 15 (baseline; BASE), day 22 (challenge day; CHAL), and on day 29 (168th hour post acid challenge or recovery, REC) of each period. Over the study, duration of pH acidosis) was less for ADY and KDY than CTRL, with ADY less than KDY. No treatment effects were observed on relative abundance of ruminal bacteria, but the day effect was significant. The RPS of lactate producers and utilizers was greater while RPS of fibrolytic bacteria was lower during CHAL than BASE and REC. Yeast supplementation, irrespective of its viability, showed beneficial effects on ruminal pH variables in animals more susceptible to acidosis. Rumen microbial population was altered with the induction of severe acidosis. Most of the changes reverted back to baseline values during the recovery phase. Yeast supplementation reduced subacute rumen acidosis in the most susceptible cattle, but failed to attenuate severe acidosis induced by a grain challenge. The study provided valuable insight into the mechanism by which acidosis affects cattle performance. Individual animal variation in ruminal fermentation partly explained the variability in response to yeast supplementation in the study. © 2017 Her Majesty the Queen in Right of Canada. Journal of Applied Microbiology © 2017 The

  10. Factors affecting the numbers of expected viable lactic acid bacteria in inoculant applicator tanks.

    Science.gov (United States)

    Windle, M C; Kung, L

    2016-11-01

    The application of correct numbers of viable microorganisms to forages at the time of ensiling is one of the most important factors affecting the probability of a beneficial effect from an inoculant. The objective of this study was to determine relationships between numbers of expected lactic acid bacteria (LAB) from silage inoculants in application tanks and various factors that might affect their viability. The pH and temperature of inoculant-water mixes were measured in applicator tanks (n=53) on farms in Wisconsin, Minnesota, South Dakota, and California during the corn harvest season of 2012. Samples were collected on-farm and plated on de Man, Rogosa, and Sharpe agar to enumerate LAB and establish the number of viable LAB (cfu/mL). Expected numbers of LAB were calculated from the minimum label guarantees for viable bacteria and mixing rates with water. In addition, the pH of the inoculant-water mixes at sampling, the ambient temperature at sampling, and the length of time that the samples had been in the tank were measured and obtained. The log difference between the measured and expected numbers of LAB was calculated and expressed as ΔM - E in log scale. Ambient temperature at sampling had no relationship with time in the tank or ΔM - E. Most (83%) of the inoculants had been mixed with water in the applicator tanks for <10h. For these samples, a negative linear correlation (R2=0.36) existed between time that the inoculant-water mixes were in the applicators tanks and ΔM - E. The pH of the inoculant-water mixes was also negatively correlated (R2=0.28) with time in the applicator tank, but pH was not related to ΔM - E. The temperatures of the inoculant-water mixtures were negatively correlated with ΔM - E (R2=0.39). Seven of 8 samples whose ΔM - E were at least -0.95 or more lower than expected (equivalent of about 1 or more log concentration less than expected) had water temperatures above 35°C. These data support our previous laboratory findings and

  11. IMPROVING CO2 EFFICIENCY FOR RECOVERING OIL IN HETEROGENEOUS RESERVOIRS

    Energy Technology Data Exchange (ETDEWEB)

    Reid B. Grigg; Robert K. Svec

    2002-12-20

    This document is the First Annual Report for the U.S. Department of Energy under contract No., a three-year contract entitled: ''Improving CO{sub 2} Efficiency for Recovering Oil in Heterogeneous Reservoirs.'' The research improved our knowledge and understanding of CO{sub 2} flooding and includes work in the areas of injectivity and mobility control. The bulk of this work has been performed by the New Mexico Petroleum Recovery Research Center, a research division of New Mexico Institute of Mining and Technology. This report covers the reporting period of September 28, 2001 and September 27, 2002. Injectivity continues to be a concern to the industry. During this period we have contacted most of the CO{sub 2} operators in the Permian Basin and talked again about their problems in this area. This report has a summary of what we found. It is a given that carbonate mineral dissolution and deposition occur in a formation in geologic time and are expected to some degree in carbon dioxide (CO{sub 2}) floods. Water-alternating-gas (WAG) core flood experiments conducted on limestone and dolomite core plugs confirm that these processes can occur over relatively short time periods (hours to days) and in close proximity to each other. Results from laboratory CO{sub 2}-brine flow experiments performed in rock core were used to calibrate a reactive transport simulator. The calibrated model is being used to estimate in situ effects of a range of possible sequestration options in depleted oil/gas reservoirs. The code applied in this study is a combination of the well known TOUGH2 simulator, for coupled groundwater/brine and heat flow, with the chemistry code TRANS for chemically reactive transport. Variability in response among rock types suggests that CO{sub 2} injection will induce ranges of transient and spatially dependent changes in intrinsic rock permeability and porosity. Determining the effect of matrix changes on CO{sub 2} mobility is crucial in

  12. CONCEIVING AND GENERATING A VIABLE COMPLEXITY MODEL FOR PERSONAL AND/OR ENTREPRENEURIAL CHANGE AND DEVELOPMENT

    Directory of Open Access Journals (Sweden)

    George Niculescu

    2012-03-01

    Full Text Available This article proposes on the one hand to do research and study – with a genuine approach, inspiredand deep-rooted in the method of the sociological school in Bucharest and creatively renewed and integrating thenewest acquisitions of complexity science – the representative intricate entity “man as an economic actor (resourceand potential within the industrial organization and corporation in mono-industrial area (region in the period inwhich the phenomenon of transition from an industrial society to knowledge society takes place; and on the otherhand our project proposes to conceive a viable model of change and career of the human resources in the ElectricPlant of Rovinari as well as to conceive and accomplish a pilot-model of a platform in order to support the efficientimplement through a research-action approach of the model of personal and/or entrepreneurship change and careerof the human resources in the Electric Plant of Rovinari.

  13. A fusion-driven subcritical system concept based on viable technologies

    Science.gov (United States)

    Wu, Y.; Jiang, J.; Wang, M.; Jin, M.; FDS Team

    2011-10-01

    A fusion-driven hybrid subcritical system (FDS) concept has been designed and proposed as spent fuel burner based on viable technologies. The plasma fusion driver can be designed based on relatively easily achieved plasma parameters extrapolated from the successful operation of existing fusion experimental devices such as the EAST tokamak in China and other tokamaks in the world, and the subcritical fission blanket can be designed based on the well-developed technologies of fission power plants. The simulation calculations and performance analyses of plasma physics, neutronics, thermal-hydraulics, thermomechanics and safety have shown that the proposed concept can meet the requirements of tritium self-sufficiency and sufficient energy gain as well as effective burning of nuclear waste from fission power plants and efficient breeding of nuclear fuel to feed fission power plants.

  14. Drug delivery interfaces in the 21st century: from science fiction ideas to viable technologies.

    Science.gov (United States)

    Chertok, Beata; Webber, Matthew J; Succi, Marc D; Langer, Robert

    2013-10-07

    Early science fiction envisioned the future of drug delivery as targeted micrometer-scale submarines and "cyborg" body parts. Here we describe the progression of the field toward technologies that are now beginning to capture aspects of this early vision. Specifically, we focus on the two most prominent types of systems in drug delivery: the intravascular micro/nano drug carriers for delivery to the site of pathology and drug-loaded implantable devices that facilitate release with the predefined kinetics or in response to a specific cue. We discuss the unmet clinical needs that inspire these designs, the physiological factors that pose difficult challenges for their realization, and viable technologies that promise robust solutions. We also offer a perspective on where drug delivery may be in the next 50 years based on expected advances in material engineering and in the context of future diagnostics.

  15. An online listserv for nurse practitioners: a viable venue for continuous nursing professional development?

    Science.gov (United States)

    Hew, Khe Foon; Hara, Noriko

    2008-05-01

    This study reports the results of a qualitative study involving a large and longstanding online nurse listserv in the United States. A sample of 27 critical care and advanced-practice nurse practitioners was interviewed using semi-structured individual interviews. This study found evidence that participation in an online listserv offers a viable avenue for the continuous professional development of nurses by providing nurses the opportunity to make more informed decisions about their professional practice and keeping abreast with up-to-date changes in their specialty areas when they shared knowledge with one another. Follow-up interviews with 10 nurses who frequently shared their knowledge revealed six motivators that helped promote knowledge sharing: (a) reciprocity, (b) collectivism, (c) personal gain, (d) respectful environment, (e) altruism, and (f) technology. Implications for sustaining knowledge sharing in an online listserv are discussed. The finding will inform educators and administrators who support continuing education and professional development of healthcare professionals.

  16. Environmental assessment and viable interdependence: the Great Whale River case in northern Quebec (First Nations)

    Energy Technology Data Exchange (ETDEWEB)

    Mulvihill, P. R.

    1997-12-31

    This study is based on the belief that environmental assessment (EA) can be supportive of viable interdependence between regions and cultures. The central focus is on the scoping stage of the EA conducted for the proposed Great Whale hydroelectric project in northern Quebec. The evaluative framework consists of 16 criteria divided into three interrelated categories, i.e. substantive, general process-oriented and specific process-oriented. The specific process-oriented criteria constitute the primary analytical focus and are the subject of five separate sub-analysis, which reveal various strengths and weaknesses in the performance of the case study. It was concluded that environmental assessment in an intercultural setting is largely within the control of EA panels and the key shortcoming of the process, namely the lack of dialogue between the proponents and the intervenors, could be addressed by making public hearings more dynamic and interactive.

  17. Is Telemental Health Services a Viable Alternative to Traditional Psychotherapy for Deaf Individuals?

    Science.gov (United States)

    Crowe, Teresa V

    2017-02-01

    Access to mental health treatment is a vital part of a comprehensive health care plan. Deaf individuals often have difficulty accessing mental health services. Four-hundred twenty-two participants completed an anonymous questionnaire about their perspectives of telemental health services for deaf individuals. Results showed that several variables, such as if the participant was unable to receive another type of psychotherapy and whether the therapist was ASL-fluent, were significantly related to whether the respondent would use TMH. The participants reported that TMH services are a viable option for treating a variety of mental health issues. Telemental health services can act as a bridge between consumers of mental health care and their providers allowing accessible and equitable healthcare opportunities.

  18. Drug Delivery Interfaces in the 21st Century: From Science Fiction Ideas to Viable Technologies

    Science.gov (United States)

    Chertok, Beata; Webber, Matthew J.; Succi, Marc D.; Langer, Robert S.

    2013-01-01

    Early science fiction envisioned the future of drug delivery as targeted micron-scale submarines and ‘Cyborg’ body parts. Here we describe the progression of the field toward technologies that are now beginning to capture aspects of this early vision. Specifically, we focus on the two most prominent types of systems in drug delivery – the intravascular micro/nano drug carriers for delivery to the site of pathology and drug-loaded implantable devices that facilitate release with the pre-defined kinetics or in response to a specific cue. We discuss the unmet clinical needs that inspire these designs, the physiological factors that pose difficult challenges for their realization, and viable technologies that promise robust solutions. We also offer a perspective on where drug delivery may be in the next 50 years based on expected advances in material engineering and in the context of future diagnostics. PMID:23915375

  19. Minimum Viable Product and the Importance of Experimentation in Technology Startups

    Directory of Open Access Journals (Sweden)

    Dobrila Rancic Moogk

    2012-03-01

    Full Text Available Entrepreneurs are often faced with limited resources in their quest to commercialize new technology. This article presents the model of a lean startup, which can be applied to an organization regardless of its size or environment. It also emphasizes the conditions of extreme uncertainty under which the commercialization of new technology is carried out. The lean startup philosophy advocates efficient use of resources by introducing a minimum viable product to the market as soon as possible in order to test its value and the entrepreneur’s growth projections. This testing is done by running experiments that examine the metrics relevant to three distinct types of the growth. These experiments bring about accelerated learning to help reduce the uncertainty that accompanies commercialization projects, thereby bringing the resulting new technology to market faster.

  20. FREE OPEN SOURCE AND SOFTWARE IN THE TEACHING OF CAT TOOLS: OMEGAT, A VIABLE ALTERNATIVE

    Directory of Open Access Journals (Sweden)

    Adauto Lúcio Caetano Villela

    2016-12-01

    Full Text Available The teaching of Computer Aided Translation (CAT tools is essential to all courses aimed at preparing students for the effective exercise of this profession, in particular those students who will translate technical texts, especially in the field of localization. As an alternative to the paid commercial software which dominate the translation industry (such as SDL Trados Studio, Wordfast Pro and MemoQ, there are free proprietary softwares (such as Wordfast Anywhere and Google Translator Toolkit, and free open source ones (such as OmegaT and Anaphraseus. Starting from a description of CAT types and main functions, the purpose of this article is to point out, through an evaluative-comparative analysis based on a research and on a comparison between OmegaT, free proprietary softwares and others open source softwares, why and for which situations OmegaT is a viable alternative for the teaching of CAT tools in higher education.

  1. Development of a Commercially Viable, Modular Autonomous Robotic Systems for Converting any Vehicle to Autonomous Control

    Science.gov (United States)

    Parish, David W.; Grabbe, Robert D.; Marzwell, Neville I.

    1994-01-01

    A Modular Autonomous Robotic System (MARS), consisting of a modular autonomous vehicle control system that can be retrofit on to any vehicle to convert it to autonomous control and support a modular payload for multiple applications is being developed. The MARS design is scalable, reconfigurable, and cost effective due to the use of modern open system architecture design methodologies, including serial control bus technology to simplify system wiring and enhance scalability. The design is augmented with modular, object oriented (C++) software implementing a hierarchy of five levels of control including teleoperated, continuous guidepath following, periodic guidepath following, absolute position autonomous navigation, and relative position autonomous navigation. The present effort is focused on producing a system that is commercially viable for routine autonomous patrolling of known, semistructured environments, like environmental monitoring of chemical and petroleum refineries, exterior physical security and surveillance, perimeter patrolling, and intrafacility transport applications.

  2. Heterogeneity, politics of ethnicity, and multiculturalism What is a viable framework for Indonesia?

    Directory of Open Access Journals (Sweden)

    Thung Ju Lan

    2011-10-01

    Full Text Available Indonesia is a plural society that consists of several hundred ethnic and sub-ethnic groups. One of its generic characteristics is heterogeneity. In the last ten years after the implementation of regional autonomy, we have witnessed the emergence of strong ethnic and religiously flavoured local identity politics in various places in Indonesia that created open and vicious conflicts. This periodical violence exploded especially during the election of district and provincial heads. The intimate relation multiculturalism, with the actual political praxis of everyday life as an alternative to the existing paradigm of the “homogenization” of nationhood, has not been discussed. I believe it is time to discuss the strategic junctures between heterogeneity, politics of ethnicity (and religion and multiculturalism as well as their possible realization at the local and national levels in order to find a viable framework for a future Indonesia.

  3. Fern spore longevity in saline water: can sea bottom sediments maintain a viable spore bank?

    Directory of Open Access Journals (Sweden)

    G Arjen de Groot

    Full Text Available Freshwater and marine sediments often harbor reservoirs of plant diaspores, from which germination and establishment may occur whenever the sediment falls dry. Therewith, they form valuable records of historical inter- and intraspecific diversity, and are increasingly exploited to facilitate diversity establishment in new or restored nature areas. Yet, while ferns may constitute a considerable part of a vegetation's diversity and sediments are known to contain fern spores, little is known about their longevity, which may suffer from inundation and--in sea bottoms--salt stress. We tested the potential of ferns to establish from a sea or lake bottom, using experimental studies on spore survival and gametophyte formation, as well as a spore bank analysis on sediments from a former Dutch inland sea. Our experimental results revealed clear differences among species. For Asplenium scolopendrium and Gymnocarpium dryopteris, spore germination was not affected by inundated storage alone, but decreased with rising salt concentrations. In contrast, for Asplenium trichomanes subsp. quadrivalens germination decreased following inundation, but not in response to salt. Germination rates decreased with time of storage in saline water. Smaller and less viable gametophytes were produced when saline storage lasted for a year. Effects on germination and gametophyte development clearly differed among genotypes of A. scolopendrium. Spore bank analyses detected no viable spores in marine sediment layers. Only two very small gametophytes (identified as Thelypteris palustris via DNA barcoding emerged from freshwater sediments. Both died before maturation. We conclude that marine, and likely even freshwater sediments, will generally be of little value for long-term storage of fern diversity. The development of any fern vegetation on a former sea floor will depend heavily on the deposition of spores onto the drained land by natural or artificial means of dispersal.

  4. Fern spore longevity in saline water: can sea bottom sediments maintain a viable spore bank?

    Science.gov (United States)

    de Groot, G Arjen; During, Heinjo

    2013-01-01

    Freshwater and marine sediments often harbor reservoirs of plant diaspores, from which germination and establishment may occur whenever the sediment falls dry. Therewith, they form valuable records of historical inter- and intraspecific diversity, and are increasingly exploited to facilitate diversity establishment in new or restored nature areas. Yet, while ferns may constitute a considerable part of a vegetation's diversity and sediments are known to contain fern spores, little is known about their longevity, which may suffer from inundation and--in sea bottoms--salt stress. We tested the potential of ferns to establish from a sea or lake bottom, using experimental studies on spore survival and gametophyte formation, as well as a spore bank analysis on sediments from a former Dutch inland sea. Our experimental results revealed clear differences among species. For Asplenium scolopendrium and Gymnocarpium dryopteris, spore germination was not affected by inundated storage alone, but decreased with rising salt concentrations. In contrast, for Asplenium trichomanes subsp. quadrivalens germination decreased following inundation, but not in response to salt. Germination rates decreased with time of storage in saline water. Smaller and less viable gametophytes were produced when saline storage lasted for a year. Effects on germination and gametophyte development clearly differed among genotypes of A. scolopendrium. Spore bank analyses detected no viable spores in marine sediment layers. Only two very small gametophytes (identified as Thelypteris palustris via DNA barcoding) emerged from freshwater sediments. Both died before maturation. We conclude that marine, and likely even freshwater sediments, will generally be of little value for long-term storage of fern diversity. The development of any fern vegetation on a former sea floor will depend heavily on the deposition of spores onto the drained land by natural or artificial means of dispersal.

  5. Sensitive and specific detection of viable Mycobacterium avium subsp. paratuberculosis in raw milk by the peptide-mediated magnetic separation-phage assay.

    Science.gov (United States)

    Foddai, A C G; Grant, I R

    2017-05-01

    To validate an optimized peptide-mediated magnetic separation (PMS)-phage assay for detection of viable Mycobacterium avium subsp. paratuberculosis (MAP) in milk. Inclusivity, specificity and limit of detection 50% (LOD50 ) of the optimized PMS-phage assay were assessed. Plaques were obtained for all 43 MAP strains tested. Of 12 other Mycobacterium sp. tested, only Mycobacterium bovis BCG produced small numbers of plaques. LOD50 of the PMS-phage assay was 0·93 MAP cells per 50 ml milk, which was better than both PMS-qPCR and PMS-culture. When individual milks (n = 146) and bulk tank milk (BTM, n = 22) obtained from Johne's affected herds were tested by the PMS-phage assay, viable MAP were detected in 31 (21·2%) of 146 individual milks and 13 (59·1%) of 22 BTM, with MAP numbers detected ranging from 6-948 plaque-forming-units per 50 ml milk. PMS-qPCR and PMS-MGIT culture proved to be less sensitive tests than the PMS-phage assay. The optimized PMS-phage assay is the most sensitive and specific method available for the detection of viable MAP in milk. Further work is needed to streamline the PMS-phage assay, because the assay's multistep format currently makes it unsuitable for adoption by the dairy industry as a screening test. The inclusivity (ability to detect all MAP strains), specificity (ability to detect only MAP) and detection sensitivity (ability to detect low numbers of MAP) of the optimized PMS-phage assay have been comprehensively demonstrated for the first time. © 2017 The Society for Applied Microbiology.

  6. New procedure for recovering extra- and intracellular DNA from marine sediment samples

    Science.gov (United States)

    Alawi, M.; Kallmeyer, J.

    2012-12-01

    Extracellular DNA (eDNA) is a ubiquitous biological compound in aquatic sediment and soil. Despite major methodological advances, analysis of DNA from sediment is still technically challenging, not just because of the co-elution of inhibitory substances, but also due to co-elution of extracellular DNA, which potentially leads to an overestimate of the actual diversity. Previous studies suggested that eDNA might play an important role in biogeochemical element cycling, horizontal gene transfer and stabilization of biofilm structures. Several protocols based on the precipitation of eDNA e.g. with CTAB and ethanol have already been published. However, using these methods we did not succeed in quantifying very low amounts of eDNA (e.g. applications like PCR can be performed. To evaluate the new extraction method two sediments with rather opposing composition were analyzed. Sediment from the South Pacific Gyre, the most oligotrophic oceanic region on earth and organic-rich Baltic Sea sediment (Northern Germany) were processed. Using this new procedure high purity genomic iDNA and eDNA with a molecular size range between 20 bp and 50k bp can be simultaneously recovered even from very oligotrophic sediment with very low cell abundances. The main fraction of recovered eDNA was suitable for downstream applications like PCR and had a molecular size that indicates minimal shearing. Despite about two decades of research many questions about deep subsurface life remain unanswered. The fact that microbes can be found even in deep oligotrophic marine sediment raises the fundamental questions of the types and availability of substrates and their biogeochemical cycling. This is the first study that provides evidence that eDNA is an important potential substrate for microorganisms in the deep biosphere. Also, our results show a link between cell counts and eDNA content, indicating that the eDNA pool in the investigated sediment consist mainly of microbial DNA. Comparative sequence

  7. Inactivation of the gene katA or sodA affects the transient entry into the viable but non-culturable response of Staphylococcus aureus in natural seawater at low temperature.

    Science.gov (United States)

    Masmoudi, Salma; Denis, Michel; Maalej, Sami

    2010-12-01

    We have investigated the fate of Staphylococcus aureus by starving the cells and maintaining them in natural seawater at 22 and 4 °C. At 22 °C, cells developed a long-term survival state where about 0.037% of the initial population remained culturable over more than 7 months, whereas at 4 °C, bacteria lost culturability and transiently entered into the viable but non-culturable state (VBNC). However, after 22 days of entry into the VBNC state, the number of viable cells detected via the direct viable count method decreased significantly. We show here that mutational inactivation of catalase (KatA) or superoxide dismutase (SodA) rendered strains hypersensitive to seawater stress at 4 °C and consequently, part of the seawater lethality on S. aureus at low temperature is mediated by reactive oxygen species (ROS) during microcosm-survival process. Shifting the temperature from 4 to 22 °C of totally non-culturable wild-type cells induced a partial recovery of the population. However, deficiencies in catalase or superoxide dismutase prevent resuscitation ability. Copyright © 2010 Elsevier Ltd. All rights reserved.

  8. From a contractors perspective: what is needed to recover used CCA treated lumber?

    Science.gov (United States)

    Delton R. Alderman; Robert L. Smith; Philip A. Araman

    2003-01-01

    The rationale of this research was to discern the factors that currently are preventing contractors from recovering and recycling spent chromated copper arsenate (CCA) lumber. This study examines the effects of evaluations and beliefs, subjective norms, perceived behavioral control, and awareness of the contractor's decision intention to recover used CCA lumber....

  9. EXPERIMENTAL INVESTIGATION OF PIC FORMATION DURING THE INCINERATION OF RECOVERED CFC-11

    Science.gov (United States)

    The report gives results of an investigation of the formation of products of incomplete combustion (PICS) during "recovered" trichlorofluoromethane (CFC-11) incineration. Tests involved burning the recovered CFC-11 in a propane gas flame. combustion gas samples were taken and an...

  10. 40 CFR Appendix A to Subpart B of... - Standard for Recycle/Recover Equipment

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 17 2010-07-01 2010-07-01 false Standard for Recycle/Recover Equipment... Conditioners Pt. 82, Subpt. B, App. A Appendix A to Subpart B of Part 82—Standard for Recycle/Recover Equipment... the ozone layer, recycle of CFC-12 (R-12) used in mobile air-conditioning systems is required to...

  11. FEASIBILITY OF THE AEROSOL-TO-LIQUID PARTICLE EXTRACTION SYSTEM (ALPES) FOR COLLECTION OF VIABLE FRANCISELLA SP.

    Energy Technology Data Exchange (ETDEWEB)

    Heitkamp, M

    2006-08-07

    the final trial runs with F. tularensis LVS to further reduce the levels of microbial background. Results from trials with F. tularensis LVS showed about a 1-log loss decrease in CFUs after 24 h, but maintained final cell concentrations in the range of 10{sup 3}-10{sup 4} CFU/L. These results indicate that the ALPES maintains acceptable viability of Francisella sp. in PBS buffer for up to 24 h and is a promising technology for the collection of viable airborne Francisella or Francisella-related cultures which may be observed at Biowatch monitoring sites in the Houston area and elsewhere.

  12. Methanogenesis in membraneless microbial electrolysis cells.

    Science.gov (United States)

    Clauwaert, Peter; Verstraete, Willy

    2009-04-01

    Operation of microbial electrolysis cells (MECs) without an ion exchange membrane could help to lower the construction costs while lowering the ohmic cell resistance and improving MEC conversion rates by minimizing the pH gradient between anode and cathode. In this research, we demonstrate that membraneless MECs with plain graphite can be operated for methane production without pH adjustment and that the ohmic cell resistance could be lowered with approximately 50% by removing the cation exchange membrane. As a result, the current production increased from 66 +/- 2 to 156 +/- 1 A m(-3) MEC by removing the membrane with an applied voltage of -0.8 V. Methane was the main energetic product despite continuous operation under carbonate-limited and slightly acidified conditions (pH 6.1-6.2). Our results suggest that continuous production of hydrogen in membraneless MECs will be challenging since methane production might not be avoided easily. The electrical energy invested was not always completely recovered under the form of an energy-rich biogas; however, our results indicate that membraneless MECs might be a viable polishing step for the treatment of the effluent of anaerobic digesters as methane was produced under low organic loading conditions and at room temperature.

  13. New Insights into the Formation of Viable but Nonculturable Escherichia coli O157:H7 Induced by High-Pressure CO2

    Directory of Open Access Journals (Sweden)

    Feng Zhao

    2016-08-01

    Full Text Available The formation of viable but nonculturable (VBNC Escherichia coli O157:H7 induced by high-pressure CO2 (HPCD was investigated using RNA sequencing (RNA-Seq transcriptomics and isobaric tag for relative and absolute quantitation (iTRAQ proteomic methods. The analyses revealed that 97 genes and 56 proteins were significantly changed upon VBNC state entry. Genes and proteins related to membrane transport, central metabolisms, DNA replication, and cell division were mainly downregulated in the VBNC cells. This caused low metabolic activity concurrently with a division arrest in cells, which may be related to VBNC state formation. Cell division repression and outer membrane overexpression were confirmed to be involved in VBNC state formation by homologous expression of z2046 coding for transcriptional repressor and ompF encoding outer membrane protein F. Upon VBNC state entry, pyruvate catabolism in the cells shifted from the tricarboxylic acid (TCA cycle toward the fermentative route; this led to a low level of ATP. Combating the low energy supply, ATP production in the VBNC cells was compensated by the degradation of l-serine and l-threonine, the increased AMP generation, and the enhanced electron transfer. Furthermore, tolerance of the cells with respect to HPCD-induced acid, oxidation, and high CO2 stresses was enhanced by promoting the production of ammonia and NADPH and by reducing CO2 production during VBNC state formation. Most genes and proteins related to pathogenicity were downregulated in the VBNC cells. This would decrease the cell pathogenicity, which was confirmed by adhesion assays. In conclusion, the decreased metabolic activity, repressed cell division, and enhanced survival ability in E. coli O157:H7 might cause HPCD-induced VBNC state formation.

  14. Rapid quantification of viable Legionella in nuclear cooling tower waters using filter cultivation, fluorescent in situ hybridization and solid-phase cytometry.

    Science.gov (United States)

    Baudart, J; Guillaume, C; Mercier, A; Lebaron, P; Binet, M

    2015-05-01

    To develop a rapid and sensitive method to quantify viable Legionella spp. in cooling tower water samples. A rapid, culture-based method capable of quantifying as few as 600 Legionella microcolonies per litre within 2 days in industrial waters was developed. The method combines a short cultivation step of microcolonies on GVPC agar plate, specific detection of Legionella cells by a fluorescent in situ hybridization (FISH) approach, and a sensitive enumeration using a solid-phase cytometer. Following optimization of the cultivation conditions, the qualitative and quantitative performance of the method was assessed and the method was applied to 262 nuclear power plant cooling water samples. The performance of this method was in accordance with the culture method (NF-T 90-431) for Legionella enumeration. The rapid detection of viable Legionella in water is a major concern to the effective monitoring of this pathogenic bacterium in the main water sources involved in the transmission of legionellosis infection (Legionnaires' disease). The new method proposed here appears to be a robust, efficient and innovative means for rapidly quantifying cultivable Legionella in cooling tower water samples within 48 h. © 2015 The Society for Applied Microbiology.

  15. The single sperm curling test, a modified hypo-osmotic swelling test, as a potential technique for the selection of viable sperm for intracytoplasmic sperm injection.

    Science.gov (United States)

    Ahmadi, A; Ng, S C

    1997-08-01

    To develop a technique for the selection of viable sperm for intracytoplasmic sperm injection (ICSI), based on the phenomenon of sperm tail curling in a hypo-osmotic environment, through modification of the hypo-osmotic swelling test. Individual sperm were exposed to single sperm curling (SSC) medium and then injected into hamster oocytes to study the effect of SSC medium on fertilization. All materials were collected from the National University Hospital in Singapore. Semen of proven donors and hamster oocytes with intact zonae were used. ICSI and the SSC test. Sperm head decondensation and male pronucleus formation. Sperm head decondensation and male pronucleus formation were present in 59.4% and 42.4%, respectively, of the oocytes injected with sperm that had been exposed to SSC medium. These rates were 70% and 48.8%, respectively, when the sperm were washed thoroughly after exposure to SSC medium. In the control group (sperm that were not exposed to SSC medium), these rates were 68.8% and 46.2%, respectively. The SSC test is useful for the selection of viable sperm for ICSI. It allows the behavioral study of a single sperm in hypo-osmotic conditions. Thorough washing of the exposed sperm is important. This procedure would be of benefit especially in testicular biopsies or very severe cases of low sperm count in which only a few sperm are found among many other cells and artifact.

  16. Demonstration of viable but nonculturable Vibrio cholerae O1 in fresh water environment of India using ciprofloxacin DFA-DVC method.

    Science.gov (United States)

    Mishra, A; Taneja, N; Sharma, M

    2011-07-01

    To demonstrate the presence of culturable and nonculturable viable pathogenic Vibrio cholerae O1 in fresh water environments of a cholera-endemic region in India. Conventional culture and ciprofloxacin DFA-DVC were utilized to investigate the existence of V. cholerae O1. We isolated pathogenic culturable V. cholerae O1 from water samples collected from cholera-affected areas. No culturable V. cholerae O1 was isolated from water and plankton samples from natural fresh water bodies. Ciprofloxacin was used for DFA-DVC as V. cholerae O1 are 100% resistant to nalidixic acid in our region. The viable but nonculturable O1 cells were demonstrated in 2.21 and 40.69% samples from natural water bodies and cholera-affected areas, respectively. Vibrio cholerae O1 VBNC could be demonstrated using modified DFA-DVC technique. Ciprofloxacin is preferable to nalidixic acid for DVC in view of existing high-level resistance to nalidixic acid in cholera-endemic areas. We endorse that for public health surveillance, cholera outbreak investigation and disease control water samples in addition to culture should be tested for V. cholerae using DFA-DVC. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

  17. Expression of proposed implantation marker genes CDX2 and HOXB7 in the blastocyst does not distinguish viable from non-viable human embryos

    DEFF Research Database (Denmark)

    Kirkegaard, Kirstine; Hindkjær, Johnny Juhl; Ingerslev, Hans Jakob

    2012-01-01

    Introduction: Selection of the most competent embryo challenges the credo of promoting single embryo transfer (SET) while maintaining high pregnancy rates, thus intensifying the need to introduce techniques that can improve embryo selection. Recent studies have demonstrated that blastocyst gene...... after oocyte retrieval. Immediately following biopsy, cells were lysed and reverse transcriptase PCR (RT-PCR) was carried out directly on the cell lysate using Superscript® III First Strand Synthesis kit (Invitrogen) thus avoiding introducing RNA amplification steps. As part of the study, the expression...... of 15 key genes associated with developmental competence in animals were evaluated in high quality human embryos with monogenic or chromosomal disorders from a pre-implantation genetic disorder program. Triplicate cDNA amplifications for quantitative (q) RT-PCR were performed using pre-designed gene...

  18. Improving the Performance of Somatic Mutation Identification by Recovering Circulating Tumor DNA Mutations.

    Science.gov (United States)

    Fu, Yu; Jovelet, Cécile; Filleron, Thomas; Pedrero, Marion; Motté, Nelly; Boursin, Yannick; Luo, Yufei; Massard, Christophe; Campone, Mario; Levy, Christelle; Diéras, Véronique; Bachelot, Thomas; Garrabey, Julie; Soria, Jean-Charles; Lacroix, Ludovic; André, Fabrice; Lefebvre, Celine

    2016-10-15

    DNA extracted from cancer patients' whole blood may contain somatic mutations from circulating tumor DNA (ctDNA) fragments. In this study, we introduce cmDetect, a computational method for the systematic identification of ctDNA mutations using whole-exome sequencing of a cohort of tumor and corresponding peripheral whole-blood samples. Through the analysis of simulated data, we demonstrated an increase in sensitivity in calling somatic mutations by combining cmDetect to two widely used mutation callers. In a cohort of 93 breast cancer metastatic patients, cmDetect identified ctDNA mutations in 54% of the patients and recovered somatic mutations in cancer genes EGFR, PIK3CA, and TP53 We further showed that cmDetect detected ctDNA in 89% of patients with confirmed mutated cell-free tumor DNA by plasma analyses (n = 9) within 46 pan-cancer patients. Our results prompt immediate consideration of the use of this method as an additional step in somatic mutation calling using whole-exome sequencing data with blood samples as controls. Cancer Res; 76(20); 5954-61. ©2016 AACR. ©2016 American Association for Cancer Research.

  19. Methotrexate encephalopathy: Two cases in adult cancer patients, who recovered with pathophysiologically based therapy

    Directory of Open Access Journals (Sweden)

    Shodeinde A Coker

    2017-05-01

    Full Text Available Background/Objectives: Neurotoxicity is a serious and sometimes fatal adverse effect that can occur following methotrexate treatment. We describe two adult patients with hematological malignancies with methotrexate encephalopathy who recovered with dextromethorphan therapy. Results: Case 1: A 24-year-old male with acute lymphoblastic leukemia developed the acute onset of bilateral facial weakness and slurred speech after his first treatment with high-dose intravenous methotrexate. The clinical scenario and a head magnetic resonance imaging supported a diagnosis of methotrexate encephalopathy. Treatment with dextromethorphan was coincident with recovery. Case 2: A 65-year-old female with recurrent diffuse large B-cell lymphoma was treated with high-dose intravenous methotrexate. Two weeks after a cycle, she developed hypoactive delirium, marked lethargy, ocular ataxia, and a right-sided facial weakness. Within 2 days of starting dextromethorphan, there was improvement with clinical recovery. Conclusions: These two cases suggest that N-methyl d-aspartate receptor activation by homocysteine may play an important role in the pathogenesis of methotrexate neurotoxicity.

  20. High-Throughput Sequencing of Viable Microbial Communities in Raw Pork Subjected to a Fast Cooling Process.

    Science.gov (United States)

    Yang, Chao; Che, You; Qi, Yan; Liang, Peixin; Song, Cunjiang

    2017-01-01

    This study aimed to investigate the effect of the fast cooling process on the microbiological community in chilled fresh pork during storage. We established a culture-independent method to study viable microbes in raw pork. Tray-packaged fresh pork and chilled fresh pork were completely spoiled after 18 and 49 d in aseptic bags at 4 °C, respectively. 16S/18S ribosomal RNAs were reverse transcribed to cDNA to characterize the activity of viable bacteria/fungi in the 2 types of pork. Both cDNA and total DNA were analyzed by high-throughput sequencing, which revealed that viable Bacteroides sp. were the most active genus in rotten pork, although viable Myroides sp. and Pseudomonas sp. were also active. Moreover, viable fungi were only detected in chilled fresh pork. The sequencing results revealed that the fast cooling process could suppress the growth of microbes present initially in the raw meat to extend its shelf life. Our results also suggested that fungi associated with pork spoilage could not grow well in aseptic tray-packaged conditions. © 2016 Institute of Food Technologists®.

  1. Arginase treatment prevents the recovery of canine lymphoma and osteosarcoma cells resistant to the toxic effects of prolonged arginine deprivation.

    Directory of Open Access Journals (Sweden)

    James W Wells

    Full Text Available Rapidly growing tumor cells require a nutrient-rich environment in order to thrive, therefore, restricting access to certain key amino acids, such as arginine, often results in the death of malignant cells, which frequently display defective cell cycle check-point control. Healthy cells, by contrast, become quiescent and remain viable under arginine restriction, displaying full recovery upon return to arginine-rich conditions. The use of arginase therapy to restrict available arginine for selectively targeting malignant cells is currently under investigation in human clinical trials. However, the suitability of this approach for veterinary uses is unexplored. As a prelude to in vivo studies in canine malignancies, we examined the in vitro effects of arginine-deprivation on canine lymphoid and osteosarcoma cell lines. Two lymphoid and 2 osteosarcoma cell lines were unable to recover following 6 days of arginine deprivation, but all remaining cell lines displayed full recovery upon return to arginine-rich culture conditions. These remaining cell lines all proved susceptible to cell death following the addition of arginase to the cultures. The lymphoid lines were particularly sensitive to arginase, becoming unrecoverable after just 3 days of treatment. Two of the osteosarcoma lines were also susceptible over this time-frame; however the other 3 lines required 6-8 days of arginase treatment to prevent recovery. In contrast, adult progenitor cells from the bone marrow of a healthy dog were able to recover fully following 9 days of culture in arginase. Over 3 days in culture, arginase was more effective than asparaginase in inducing the death of lymphoid lines. These results strongly suggest that short-term arginase treatment warrants further investigation as a therapy for lymphoid malignancies and osteosarcomas in dogs.

  2. A Preliminary Study Examining Women's Physical Health and Nonmedical Prescription Opioid Use in a Recovering Framework.

    Science.gov (United States)

    Marks, Katherine R; Leukefeld, Carl G

    2017-03-01

    There is a growing need for research on women recovering from nonmedical prescription opioid use. A paradigm shift is to focus on wellness and factors that support, rather than hinder, recovering women. For example, although physical health is impacted by nonmedical prescription opioid use, improved health may be a strength-based factor among recovering women. The aim of this preliminary study was to examine physical health in a recovering framework as women begin recovering from nonmedical prescription opiate use. This study conducted secondary data analysis using self-report data from Kentucky's Targeted Assessment Program (TAP). TAP assesses and provides pretreatment for problems impeding work or interfering with parental responsibilities. Data included 1247 adult women reporting a history of nonmedical prescription opioid use and receiving pretreatment. Bivariate and logistic regression analyses examined whether physical health and access to health services at baseline were associated with substance-related behavior change upon completion of pretreatment. Women reporting better physical health and fewer days with no activities due to health barriers were more likely to engage in pretreatment substance-related behavior change [χ2(8) = 34.6, p medical providers should be incorporated into care to meet the need. Future research should continue to examine factors that support recovering women, using a recovering framework.

  3. A direct technique for magnetic functionalization of living human cells.

    Science.gov (United States)

    Dzamukova, Maria R; Zamaleeva, Alsu I; Ishmuchametova, Dilara G; Osin, Yuri N; Kiyasov, Andrey P; Nurgaliev, Danis K; Ilinskaya, Olga N; Fakhrullin, Rawil F

    2011-12-06

    Functionalized living cells are regarded as effective tools in directed cell delivery and tissue engineering. Here we report the facile functionalization of viable isolated HeLa cells with superparamagnetic cationic nanoparticles via a single-step biocompatible process. Nanoparticles are localized on the cellular membranes and do not penetrate into the cytoplasm. The magnetically responsive cells are viable and able to colonize and grow on substrates. Magnetically facilitated microorganization of functionalized cells into viable living clusters is demonstrated. We believe that the technique described here may find a number of potential applications in cell-based therapies and in development of whole-cell biosensors. © 2011 American Chemical Society

  4. Graphene oxide and carboxylated graphene oxide: Viable two-dimensional nanolabels for lateral flow immunoassays.

    Science.gov (United States)

    Yu, Li; Li, Peiwu; Ding, Xiaoxia; Zhang, Qi

    2017-04-01

    Graphene oxide (GO) and carboxylated GO were used as labels for lateral flow immunoassays, instead of the conventionally used colloidal gold and colored latex labels. A sensor is demonstrated that enables fast screening for aflatoxin B 1 (AFB 1 ) as a model analyte using the antibody-GO complex as the recognition element. The visual limit of detection and cut-off value for AFB 1 are 0.3 and 1ng/mL, respectively. It is shown that GO and carboxylated GO are viable black labels for use in lateral flow assays, one typical advantage being the saving cost (compared to the use of colloidal gold). Qualitative results are achieved within 15min, and the analytical results were in good agreement with the reference LC MS/MS method. The method was successfully applied to the on-site determination of AFB 1 in agricultural products. In our perception, it opens new possibilities for the screening of other toxins by lateral flow immunoassays using GO and carboxylated GO as labels. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Meatal Mobilization and Glanuloplasty: A Viable Option for Coronal and Glanular Hypospadias Repair.

    Science.gov (United States)

    Moradi, Mahmoudreza; Kazemzadeh, Babak; Hood, Brandy; Rezaee, Haress; Kaseb, Kaveh

    2016-08-01

    To present the meatal mobilization with glanuloplasty inclusive (MMGPI) modification of meatal advancement and glanuloplasty inclusive. A total of 120 patients with anterior hypospadias underwent MMGPI between September 2008 and October 2014 at Kermanshah University of Medical Sciences. Satisfactory outcomes were defined as continuous straight urinary flow and catheterization of new meatus without difficulty. Cosmetic outcomes were considered acceptable if patients maintained a slit-like meatus at the glanular tip. Patients were examined at 1 week, 1, 3, 6, 12, and 24 months. The 120 patients with anterior hypospadias underwent MMGPI. There were no hematoma, meatal necrosis, or other early complications. In patients with glanular hypospadias, there were no meatal regressions or stenosis, all voiding patterns were normal, and all patients maintained a slit-like meatus at the glanular tip. Two patients with coronal hypospadias had meatal stenosis and 2 patients had meatal regression. Five patients with sub-coronal hypospadias had 2-mm meatal regression with downward sloping urinary stream, and 2 patients had meatal stenosis. In all, meatus remained distal to the preoperative meatus with no necrosis. Small sample size was the major limitation of this study. MMGPI represents a viable option for glanular and coronal hypospadias repair. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. A clinically viable capsule endoscopy video analysis platform for automatic bleeding detection

    Science.gov (United States)

    Yi, Steven; Jiao, Heng; Xie, Jean; Mui, Peter; Leighton, Jonathan A.; Pasha, Shabana; Rentz, Lauri; Abedi, Mahmood

    2013-02-01

    In this paper, we present a novel and clinically valuable software platform for automatic bleeding detection on gastrointestinal (GI) tract from Capsule Endoscopy (CE) videos. Typical CE videos for GI tract run about 8 hours and are manually reviewed by physicians to locate diseases such as bleedings and polyps. As a result, the process is time consuming and is prone to disease miss-finding. While researchers have made efforts to automate this process, however, no clinically acceptable software is available on the marketplace today. Working with our collaborators, we have developed a clinically viable software platform called GISentinel for fully automated GI tract bleeding detection and classification. Major functional modules of the SW include: the innovative graph based NCut segmentation algorithm, the unique feature selection and validation method (e.g. illumination invariant features, color independent features, and symmetrical texture features), and the cascade SVM classification for handling various GI tract scenes (e.g. normal tissue, food particles, bubbles, fluid, and specular reflection). Initial evaluation results on the SW have shown zero bleeding instance miss-finding rate and 4.03% false alarm rate. This work is part of our innovative 2D/3D based GI tract disease detection software platform. While the overall SW framework is designed for intelligent finding and classification of major GI tract diseases such as bleeding, ulcer, and polyp from the CE videos, this paper will focus on the automatic bleeding detection functional module.

  7. Determining size and dispersion of minimum viable populations for land management planning and species conservation

    Science.gov (United States)

    Lehmkuhl, John F.

    1984-03-01

    The concept of minimum populations of wildlife and plants has only recently been discussed in the literature. Population genetics has emerged as a basic underlying criterion for determining minimum population size. This paper presents a genetic framework and procedure for determining minimum viable population size and dispersion strategies in the context of multiple-use land management planning. A procedure is presented for determining minimum population size based on maintenance of genetic heterozygosity and reduction of inbreeding. A minimum effective population size ( N e ) of 50 breeding animals is taken from the literature as the minimum shortterm size to keep inbreeding below 1% per generation. Steps in the procedure adjust N e to account for variance in progeny number, unequal sex ratios, overlapping generations, population fluctuations, and period of habitat/population constraint. The result is an approximate census number that falls within a range of effective population size of 50 500 individuals. This population range defines the time range of short- to long-term population fitness and evolutionary potential. The length of the term is a relative function of the species generation time. Two population dispersion strategies are proposed: core population and dispersed population.

  8. Exploring viable vacua of the Z{sub 3}-symmetric NMSSM

    Energy Technology Data Exchange (ETDEWEB)

    Beuria, Jyotiranjan [Harish-Chandra Research Institute,Allahabad 211019 (India); Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai 400085 (India); Regional Centre for Accelerator-based Particle Physics, Harish-Chandra Research Institute,Allahabad 211019 (India); Chattopadhyay, Utpal [Department of Theoretical Physics, Indian Association for the Cultivation of Science, 2A & B Raja S. C. Mullick Road, Jadavpur, Kolkata 700 032 (India); Datta, AseshKrishna [Harish-Chandra Research Institute,Allahabad 211019 (India); Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai 400085 (India); Dey, Abhishek [Maulana Azad College, Government of West Bengal, 8 Rafi Ahmed Kidwai Road, Kolkata 700013 (India)

    2017-04-05

    We explore the vacua of the Z{sub 3}-symmetric Next-to-Minimal Supersymmetric Standard Model (NMSSM) and their stability by going beyond the simplistic paradigm that works with a tree-level neutral scalar potential and adheres to some specific flat directions in the field space. We work in the so-called phenomenological NMSSM (pNMSSM) scenario. Also, for our purpose, we adhere to a reasonably ‘natural’ setup by requiring |μ{sub eff}| not too large. Key effects are demonstrated by first studying the profiles of this potential under various circumstances of physical interest via a semi-analytical approach. The results thereof are compared to the ones obtained from a dedicated package like Vevacious which further incorporates the thermal effects to the potential. Regions of the pNMSSM parameter space that render the desired symmetry breaking (DSB) vacuum absolutely stable, long- or short-lived (in relation to the age of the Universe) under quantum/thermal tunneling are delineated. Regions that result in the appearance of color and charge breaking (CCB) minima are also presented. It is demonstrated that light singlet scalars along with a light LSP (lightest supersymmetric particle) having an appreciable singlino admixture are compatible with a viable DSB vacuum. Their implications for collider experiments are commented upon.

  9. The Independent Evolution Method Is Not a Viable Phylogenetic Comparative Method.

    Directory of Open Access Journals (Sweden)

    Randi H Griffin

    Full Text Available Phylogenetic comparative methods (PCMs use data on species traits and phylogenetic relationships to shed light on evolutionary questions. Recently, Smaers and Vinicius suggested a new PCM, Independent Evolution (IE, which purportedly employs a novel model of evolution based on Felsenstein's Adaptive Peak Model. The authors found that IE improves upon previous PCMs by producing more accurate estimates of ancestral states, as well as separate estimates of evolutionary rates for each branch of a phylogenetic tree. Here, we document substantial theoretical and computational issues with IE. When data are simulated under a simple Brownian motion model of evolution, IE produces severely biased estimates of ancestral states and changes along individual branches. We show that these branch-specific changes are essentially ancestor-descendant or "directional" contrasts, and draw parallels between IE and previous PCMs such as "minimum evolution". Additionally, while comparisons of branch-specific changes between variables have been interpreted as reflecting the relative strength of selection on those traits, we demonstrate through simulations that regressing IE estimated branch-specific changes against one another gives a biased estimate of the scaling relationship between these variables, and provides no advantages or insights beyond established PCMs such as phylogenetically independent contrasts. In light of our findings, we discuss the results of previous papers that employed IE. We conclude that Independent Evolution is not a viable PCM, and should not be used in comparative analyses.

  10. ATP bioluminescence rapid detection of total viable count in soy sauce.

    Science.gov (United States)

    Yan, Shou-Lei; Miao, Su-Na; Deng, Shao-Ya; Zou, Min-Juan; Zhong, Fo-Sheng; Huang, Wen-Biao; Pan, Si-Yi; Wang, Qing-Zhang

    2012-01-01

    The adenosine triphosphate (ATP) bioluminescence rapid determination method may be useful for enumerating the total viable count (TVC) in soy sauce, as it has been previously used in food and beverages for sanitation with good precision. However, many factors interfere with the correlation between total aerobic plate counts and ATP bioluminescence. This study investigated these interfering factors, including ingredients of soy sauce and bacteria at different physiological stages. Using the ATP bioluminescence method, TVC was obtained within 4 h, compared to 48 h required for the conventional aerobic plate count (APC) method. Our results also indicated a high correlation coefficient (r = 0.90) between total aerobic plate counts and ATP bioluminescence after filtration and resuscitation with special medium. The limit of quantification of the novel detection method is 100 CFU/mL; there is a good linear correlation between the bioluminescence intensity and TVC in soy sauce in the range 1 × 10(2) -3 × 10(4) CFU/mL and even wider. The method employed a luminescence recorder (Tristar LB-941) and 96-well plates and could analyse 50-100 samples simultaneously at low cost. In this study, we evaluated and eliminated the interfering factors and made the ATP bioluminescence rapid method available for enumerating TVC in soy sauce. Copyright © 2011 John Wiley & Sons, Ltd.

  11. Novel Approach for Enterocutaneous Fistula Treatment with the Use of Viable Cryopreserved Placental Membrane

    Directory of Open Access Journals (Sweden)

    Frederick Nichols

    2016-01-01

    Full Text Available Enterocutaneous fistulas (ECF are a difficult and costly surgical complication to manage. The standard treatment of nil per os (NPO and total paraenteral nutrition (TPN is not well tolerated by patients. TPN is also known for complications associated with long term central venous catheterization and for high cost of prolonged hospital stay. We present two low output ECF cases successfully treated with viable cryopreserved placental membrane (vCPM placed into the fistula tracts. One patient is a 59-year-old male with a low output ECF from a jejunostomy tube site four weeks after the surgery. The second patient is an 87-year-old male with a low output ECF following a small bowel resection secondary to a strangulated inguinal hernia. He was evaluated on day 41 after surgery. NPO and TPN for several weeks did not resolute the ECF. The fistulae were closed postoperatively in both patients with zero output on the same day after one vCPM application. On day 3 postoperatively both patients were started on clear liquid diets and subsequently advanced to regular diets. The ECF have remained resolved for over 2 months. The use of vCPM is a novel promising approach for treatment of ECF.

  12. Agouti regulation of intracellular calcium: Role in the insulin resistance of viable yellow mice

    Energy Technology Data Exchange (ETDEWEB)

    Zemel, M.B.; Kim, J.H. [Univ. of Tennessee, Knoxville, TN (United States); Woychik, R.P.; Michaud, E.J. [Oak Ridge National Laboratory, Oak Ridge, TN (United States); Hadwell, S.H.; Patel, I.R.; Wilkison, W.O. [Research Institute, Research Triangle Park, NC (United States)

    1995-05-23

    Several dominant mutations at the agouti locus in the mouse cause a syndrome of marked obesity, hyperinsulinemia, and insulin resistance. Although it is known that the agouti gene is expressed in an ectopic manner in these mutants, the precise mechanism by which the agouti gene product mediates these effects is unclear. Since intracellular Ca{sup 2+} is believed to play a role in mediating insulin action and dysregulation of Ca{sup 2+} flux is observed in diabetic animals and humans, we examined the status of intracellular Ca{sup 2+} in mice carrying the dominant agouti allele, viable yellow (A{sup vy}). We show here that in mice carrying this mutation, the intracellular free calcium concentration ([Ca{sup 2+}]{sub i}) is elevated in skeletal muscle, and the degree of elevation is closely correlated with the degree to which the mutant traits are expressed in individual animals. Moreover, we demonstrate that the agouti gene product is capable of inducing increased [Ca{sup 2+}]{sub i} in cultured and freshly isolated skeletal muscle myocytes from wild-type mice. Based on these findings, we present a model in which we propose that the agouti polypeptide promotes insulin resistance in mutant animals through its ability to increase [Ca{sup 2+}]{sub i}. 36 refs., 3 figs., 2 tabs.

  13. Low-dose dobutamine myocardial perfusion scintigraphy in the identification of viable myocardium

    Energy Technology Data Exchange (ETDEWEB)

    Moraes, Renata Freire de [Instituto Hermes Pardini, Belo Horizonte, MG (Brazil). Dept. of Molecular Imaging and Diagnosis. Div. of Nuclear Medicine; Meneghetti, Jose Claudio [Instituto do Coracao (InCor-HC/FM/USP), Sao Paulo, SP (Brazil). Unit of Nuclear Medicine and Molecular Imaging; Barroso, Adelanir Antonio, E-mail: renatafreire@yahoo.com.b [Nuclear Medcenter, Belo Horizonte, MG (Brazil)

    2010-09-15

    Objective: to evaluate the increase in specificity of dual isotope myocardial perfusion gated SPECT ({sup 99m}Tcsestamibi/thallium-201), a highly sensitive method to detect viable myocardium, with addition of data on contractile reserve simultaneously acquired by low-dose dobutamine gated SPECT, similarly to echocardiography. Materials and methods: a total of 260 myocardial segments were assessed in 13 patients with myocardial infarction referred for investigation of myocardial viability before undergoing revascularization. Cellular integrity and contractile reserve were evaluated by dual isotope perfusion myocardial gated SPECT with thallium rest and redistribution images and post-stress {sup 99m}Tc-sestamibi gated-SPECT images under basal conditions and with low-dose dobutamine. The improvement in the contractile performance detected by post-revascularization {sup 99m}Tc-sestamibi gated SPECT was the parameter considered for determining myocardial viability. For the purpose of results analysis, the functional parameters of the myocardial segments were quantified at the different phases of the study and stratified according to their viability for a later post-revascularization functional comparison. Results: in the statistical analysis, systolic wall thickening demonstrated to be a relevant parameter in the evaluation of myocardial contractile reserve by this method, with a tendency of improvement in the specificity (84%), demonstrating higher values than those observed in the literature. Conclusion: this method tends to present an effective contribution in the assessment of myocardial viability. (author)

  14. Maize Residue as a Viable Substrate for Farm Scale Cultivation of Oyster Mushroom (Pleurotus ostreatus

    Directory of Open Access Journals (Sweden)

    Abena O. Adjapong

    2015-01-01

    Full Text Available In the search for alternatives to sawdust as growing media in commercial mushroom cultivation, three organic substrates obtainable as crop residue, maize husk, maize cob, and maize stalk, with each being supplemented with rice bran, were evaluated as growth media for the oyster mushroom, Pleurotus ostreatus (Kummer. For the tested alternatives to sawdust, the harvested weight of fruiting bodies that sprouted on a kilogram maize husk media per crop (32.99 g was the highest. Sawdust media supported significantly (P<0.001 heavier fruiting bodies (42.18 than the maize residues. The peak mushroom harvests for the various substrates were obtained between the first and seventh fruiting body flushes. The biological efficiency of the substrates, which measured usable nutrients indicated that maize stalk supplemented with rice bran, was 39% compared to that of the sawdust media (60%. The maize husk media and the maize cob media had biological efficiencies of 32% and 9.5%, respectively. These results indicate that two of the tested growing media (maize stalk or husk produced mushrooms with yield characteristics that were comparable to the well-used sawdust in the cultivation of oyster mushrooms. The environmental and economic parameters involved in the use and carting of sawdust make these on-farm crop residues a viable alternative for mushroom cultivation in especially nonforest zones of Ghana.

  15. Total viable bacterial count using a real time all-fibre spectroscopic system.

    Science.gov (United States)

    Bogomolny, E; Swift, S; Vanholsbeeck, F

    2013-07-21

    Rapid, accurate and sensitive enumeration of bacterial populations in the natural environment is an essential task for many research fields. Widely used standard methods for counting bacteria such as heterotrophic plate count require 1 to 8 days of incubation time for limited accuracy, while more accurate and rapid techniques are often expensive and may require bulky equipment. In the present study, we have developed a computerized optical prototype for bacterial detection. The goal of this research was to estimate the potential of this optical system for Total Viable Bacterial Count in water. For this purpose, we tested water batches with different microbiological content. Bacterial detection was based on fluorescence enhanced by nucleic acid staining. High sensitivity was achieved by a stable diode pumped solid state laser, sensitive CCD spectrometer and in situ excitation and signal collection. The results have shown that the bacterial count from different water origins using our optical setup along with multivariate analysis presents a higher accuracy and a shorter detection time compared to standard methods. For example, in a case where the fluorescence signal is calibrated to the water batch regression line, the relative standard deviation of the optical system enumeration varies between 21 and 36%, while that of the heterotropic plate count counterpart varies between 41 and 59%. In summary, we conclude that the all-fibre optical system may offer the following advantages over conventional methods: near real time examinations, portability, sensitivity, accuracy and ability to detect 10(2) to 10(8) CFU per ml bacterial concentrations.

  16. Viable cold-tolerant iron-reducing microorganisms in geographically diverse subglacial environments

    Science.gov (United States)

    Nixon, Sophie L.; Telling, Jon P.; Wadham, Jemma L.; Cockell, Charles S.

    2017-03-01

    Subglacial environments are known to harbour metabolically diverse microbial communities. These microbial communities drive chemical weathering of underlying bedrock and influence the geochemistry of glacial meltwater. Despite its importance in weathering reactions, the microbial cycling of iron in subglacial environments, in particular the role of microbial iron reduction, is poorly understood. In this study we address the prevalence of viable iron-reducing microorganisms in subglacial sediments from five geographically isolated glaciers. Iron-reducing enrichment cultures were established with sediment from beneath Engabreen (Norway), Finsterwalderbreen (Svalbard), Leverett and Russell glaciers (Greenland), and Lower Wright Glacier (Antarctica). Rates of iron reduction were higher at 4 °C compared with 15 °C in all but one duplicated second-generation enrichment culture, indicative of cold-tolerant and perhaps cold-adapted iron reducers. Analysis of bacterial 16S rRNA genes indicates Desulfosporosinus were the dominant iron-reducing microorganisms in low-temperature Engabreen, Finsterwalderbreen and Lower Wright Glacier enrichments, and Geobacter dominated in Russell and Leverett enrichments. Results from this study suggest microbial iron reduction is widespread in subglacial environments and may have important implications for global biogeochemical iron cycling and export to marine ecosystems.

  17. Flow Cytometry Sorting to Separate Viable Giant Viruses from Amoeba Co-culture Supernatants.

    Science.gov (United States)

    Khalil, Jacques Y B; Langlois, Thierry; Andreani, Julien; Sorraing, Jean-Marc; Raoult, Didier; Camoin, Laurence; La Scola, Bernard

    2016-01-01

    Flow cytometry has contributed to virology but has faced many drawbacks concerning detection limits, due to the small size of viral particles. Nonetheless, giant viruses changed many concepts in the world of viruses, as a result of their size and hence opened up the possibility of using flow cytometry to study them. Recently, we developed a high throughput isolation of viruses using flow cytometry and protozoa co-culture. Consequently, isolating a viral mixture in the same sample became more common. Nevertheless, when one virus multiplies faster than others in the mixture, it is impossible to obtain a pure culture of the minority population. Here, we describe a robust sorting system, which can separate viable giant virus mixtures from supernatants. We tested three flow cytometry sorters by sorting artificial mixtures. Purity control was assessed by electron microscopy and molecular biology. As proof of concept, we applied the sorting system to a co-culture supernatant taken from a sample containing a viral mixture that we couldn't separate using end point dilution. In addition to isolating the quick-growing Mimivirus, we sorted and re-cultured a new, slow-growing virus, which we named "Cedratvirus." The sorting assay presented in this paper is a powerful and versatile tool for separating viral populations from amoeba co-cultures and adding value to the new field of flow virometry.

  18. In search of a viable reaction pathway in the chelation of a metallo-protein

    Science.gov (United States)

    Rose, Frisco; Hodak, Miroslav; Bernholc, Jerry

    2010-03-01

    Misfolded metallo-proteins are potential causal agents in the onset of neuro-degenerative diseases, such as Alzheimer's and Parkinson's Diseases (PD). Experimental results involving metal chelation have shown significant promise in symptom reduction and misfolding reversal. We explore, through atomistic simulations, potential reaction pathways for the chelation of Cu^2+ from the metal binding site in our representation of a partially misfolded α-synuclein, the protein implicated in PD. Our ab initio simulations use Density Functional Theory (DFT) and nudged elastic band to obtain the minimized energy coordinates of this reaction. Our simulations include ab initio water at the interaction site and in its first solvation shells, while the remainder is fully solvated with orbital-free DFT water representation [1]. Our ongoing studies of viable chelation agents include nicotine, caffeine and other potential reagents, we will review the best case agents in this presentation. [4pt] [1] Hodak M, Lu W, Bernholc J. Hybrid ab initio Kohn-Sham density functional theory/frozen-density orbital-free density functional theory simulation method suitable for biological systems. J. Chem. Phys. 2008 Jan;128(1):014101-9.

  19. Is flapless implant surgery a viable option in posterior maxilla? A review.

    Science.gov (United States)

    Doan, N; Du, Z; Crawford, R; Reher, P; Xiao, Y

    2012-09-01

    This article reviews the literature on the outcome of flapless surgery for dental implants in the posterior maxilla. The literature search was carried out in using the keywords: flapless, dental implants and maxilla. A hand search and Medline search were carried out on studies published between 1971 and 2011. The authors included research involving a minimum of 15 dental implants with a follow-up period of 1 year, an outcome measurement of implant survival, but excluded studies involving multiple simultaneous interventions, and studies with missing data. The Cochrane approach for cohort studies and Oxford Centre for Evidence-Based Medicine were applied. Of the 56 published papers selected, 14 papers on the flapless technique showed high overall implant survival rates. The prospective studies yielded 97.01% (95% CI: 90.72-99.0) while retrospective studies or case series illustrated 95.08% (95% CI: 91.0-97.93) survival. The average of intraoperative complications was 6.55% using the flapless procedure. The limited data obtained showed that flapless surgery in posterior maxilla areas could be a viable and predictable treatment method for implant placement. Flapless surgery tends to be more applicable in this area of the mouth. Further long-term clinical controlled studies are needed. Copyright © 2012 International Association of Oral and Maxillofacial Surgeons. All rights reserved.

  20. Remote sensing is a viable tool for mapping soil salinity in agricultural lands

    Directory of Open Access Journals (Sweden)

    Elia Scudiero

    2017-04-01

    Full Text Available Soil salinity negatively impacts the productivity and profitability of western San Joaquin Valley (WSJV farmland. Many factors, including drought, climate change, reduced water allocations, and land-use changes could worsen salinity conditions there, and in other agricultural lands in the state. Mapping soil salinity at regional and state levels is essential for identifying drivers and trends in agricultural soil salinity, and for developing mitigation strategies, but traditional soil sampling for salinity does not allow for accurate large-scale mapping. We tested remote-sensing modeling to map root zone soil salinity for farmland in the WSJV. According to our map, 0.78 million acres are salt affected (i.e., ECe > 4 dS/m, which represents 45% of the mapped farmland; 30% of that acreage is strongly or extremely saline. Independent validations of the remote-sensing estimations indicated acceptable to excellent correspondences, except in areas of low salinity and high soil heterogeneity. Remote sensing is a viable tool for helping landowners make decisions about land use and also for helping water districts and state agencies develop salinity mitigation strategies.