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Sample records for luciferase based viability

  1. A luciferase-based assay for rapid assessment of drug activity against Mycobacterium tuberculosis including monitoring of macrophage viability.

    Science.gov (United States)

    Larsson, Marie C; Lerm, Maria; Ängeby, Kristian; Nordvall, Michaela; Juréen, Pontus; Schön, Thomas

    2014-11-01

    The intracellular (IC) effect of drugs against Mycobacterium tuberculosis (Mtb) is not well established but increasingly important to consider when combining current and future multidrug regimens into the best possible treatment strategies. For this purpose, we developed an IC model based on a genetically modified Mtb H37Rv strain, expressing the Vibrio harvei luciferase (H37Rv-lux) infecting the human macrophage like cell line THP-1. Cells were infected at a low multiplicity of infection (1:1) and subsequently exposed to isoniazid (INH), ethambutol (EMB), amikacin (AMI) or levofloxacin (LEV) for 5days in a 96-well format. Cell viability was evaluated by Calcein AM and was maintained throughout the experiment. The number of viable H37Rv-lux was determined by luminescence and verified by a colony forming unit analysis. The results were compared to the effects of the same drugs in broth cultures. AMI, EMB and LEV were significantly less effective intracellularly (MIC90: >4mg/L, 8mg/L and 2mg/L, respectively) compared to extracellularly (MIC90: 0.5mg/L for AMI and EMB; 0.25mg/L for LEV). The reverse was the case for INH (IC: 0.064mg/L vs EC: 0.25mg/L). In conclusion, this luciferase based method, in which monitoring of cell viability is included, has the potential to become a useful tool while evaluating the intracellular effects of anti-mycobacterial drugs. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Uptake kinetics and biodistribution of C-14-D-luciferin-a radiolabeled substrate for the firefly luciferase catalyzed bioluminescence reaction : impact on bioluminescence based reporter gene imaging

    NARCIS (Netherlands)

    Berger, Frank; Paulmurugan, Ramasamy; Bhaumik, Srabani; Gambhir, Sanjiv Sam

    2008-01-01

    Purpose Firefly luciferase catalyzes the oxidative decarboxylation of D-luciferin to oxyluciferin in the presence of cofactors, producing bioluminescence. This reaction is used in optical bioluminescence-based molecular imaging approaches to detect the expression of the firefly luciferase reporter

  3. Evaluation of the Luciferase Assay-Based In Vitro Elicitation Test for Serum IgE

    Directory of Open Access Journals (Sweden)

    Ryosuke Nakamura

    2012-01-01

    Results: The area under the ROC curves was highest in the EXiLE test (0.977, followed by CAP-FEIA (0.926 and degranulation (0.810. At an optimal cutoff range (1.648-1.876 calculated from the ROC curve of the EXiLE test, sensitivity and specificity were 0.944 and 0.917, respectively. A 95% positive predictive value was given at a cutoff level of 2.054 (fold increase in luciferase expression by logistic regression analysis. Conclusions: In contrast to in vivo tests, the EXiLE test appears to be a useful tool in diagnosing patients suspected of having IgE-dependent EW allergy without the risk of severe systemic reactions.

  4. Computational analysis and functional expression of ancestral copepod luciferase.

    Science.gov (United States)

    Takenaka, Yasuhiro; Noda-Ogura, Akiko; Imanishi, Tadashi; Yamaguchi, Atsushi; Gojobori, Takashi; Shigeri, Yasushi

    2013-10-10

    We recently reported the cDNA sequences of 11 copepod luciferases from the superfamily Augaptiloidea in the order Calanoida. They were classified into two groups, Metridinidae and Heterorhabdidae/Lucicutiidae families, by phylogenetic analyses. To elucidate the evolutionary processes, we have now further isolated 12 copepod luciferases from Augaptiloidea species (Metridia asymmetrica, Metridia curticauda, Pleuromamma scutullata, Pleuromamma xiphias, Lucicutia ovaliformis and Heterorhabdus tanneri). Codon-based synonymous/nonsynonymous tests of positive selection for 25 identified copepod luciferases suggested that positive Darwinian selection operated in the evolution of Heterorhabdidae luciferases, whereas two types of Metridinidae luciferases had diversified via neutral mechanism. By in silico analysis of the decoded amino acid sequences of 25 copepod luciferases, we inferred two protein sequences as ancestral copepod luciferases. They were expressed in HEK293 cells where they exhibited notable luciferase activity both in intracellular lysates and cultured media, indicating that the luciferase activity was established before evolutionary diversification of these copepod species. © 2013.

  5. ATPase inhibitor based luciferase assay for prolonged and enhanced ATP pool measurement as an efficient fish freshness indicator.

    Science.gov (United States)

    Ranjan, Rajeev; Priyanka, B S; Thakur, M S

    2014-07-01

    The nucleotide degradation pathway in somatic cells leads to the accumulation of products such as hypoxanthine and inosine, which are commonly used as fish and meat freshness indicators. Assays based on these molecules cannot differentiate the postmortem time over a short period of time (5-10 h). Further, quantification of these degradation products is cumbersome, costly and time-consuming. For the proposed assay, optimal concentrations of 30 and 2 mM, respectively, for the ATPase inhibitors sodium orthovanadate and EDTA were found. Further, it was observed that a firefly luciferase based assay could enhance the sensitivity levels up to 165-fold at 30 °C. In addition, it was observed that the sensitivity for ATP assay was enhanced up to 60-fold even after 12 h. The limit of detection for the ATP assay was 1 pM, unlike other conventional methods, which are sensitive only up to micromolar levels. Moreover, as little as 0.044 g fish fillet was required for the assay, and no time-consuming sample preparation was necessary. Luminescence of prolonged duration was observed in harvested fish kept at -20 °C in comparison with fish kept at 4 and 30 °C, which reflects the shelf life of fish preserved at lower temperatures.

  6. In-vivo quantification of primary microRNA processing by Drosha with a luciferase based system

    International Nuclear Information System (INIS)

    Allegra, Danilo; Mertens, Daniel

    2011-01-01

    Research highlights: → Posttranscriptional regulation of miRNA processing is difficult to quantify. → Our in-vivo processing assay can quantify Drosha cleavage in live cells. → It is based on luciferase reporters fused with pri-miRNAs. → The assay validates the processing defect caused by a mutation in pri-16-1. → It is a sensitive method to quantify pri-miRNA cleavage by Drosha in live cells. -- Abstract: The RNAse III Drosha is responsible for the first step of microRNA maturation, the cleavage of primary miRNA to produce the precursor miRNA. Processing by Drosha is finely regulated and influences the amount of mature microRNA in a cell. We describe in the present work a method to quantify Drosha processing activity in-vivo, which is applicable to any microRNA. With respect to other methods for measuring Drosha activity, our system is faster and scalable, can be used with any cellular system and does not require cell sorting or use of radioactive isotopes. This system is useful to study regulation of Drosha activity in physiological and pathological conditions.

  7. Rescue the failed half-ZFN by a sensitive mammalian cell-based luciferase reporter system.

    Directory of Open Access Journals (Sweden)

    Weifeng Zhang

    Full Text Available ZFN technology is a powerful research tool and has been used for genome editing in cells lines, animals and plants. The generation of functional ZFNs for particular targets in mammalian genome is still challenging for an average research group. The modular-assembly method is relatively fast, easy-to-practice but has a high failure rate. Some recent studies suggested that a ZFP with low binding activity might be able to form a working ZFN pair with another binding active half-ZFP. In order to unveil the potential ZFP candidates among those with low binding activities, this paper established a highly sensitive mammalian cell-based transcriptional reporter system to assess the DNA binding activities of ZFPs by inserting multiple copies of ZFN target sequence fragment (TSF of an interested gene (e. g., hPGRN or hVEGF. Our results showed that this system increased the screening sensitivity up to 50-fold and markedly amplified the differences in the binding activities between different ZFPs. We also found that the targeted chromosomal gene repair efficiency of each hPGRN or hVEGF ZFN pair was in proportion with the combination of the binding activities of the ZFL (Left zinc finger and ZFR (Right zinc finger. A hPGRN ZFR with low binding ability was able to form a biological active ZFN if combined with a hPGRN ZFL with relatively high binding ability. Lastly, site-specific genome editing by hPGRN ZFNs generated by this system was confirmed by sequencing, and the PGRN knock-out cell line showed significantly decreased cell growth compared with the control. Our system will provide a valuable tool for further optimizing the nucleases with regard to specificity and cytotoxicity.

  8. Development of a luciferase-based reporter system to monitor Bifidobacterium breve UCC2003 persistence in mice

    Directory of Open Access Journals (Sweden)

    Hill Colin

    2008-09-01

    Full Text Available Abstract Background Probiotics such as bifidobacteria have been shown to maintain a healthy intestinal microbial balance and help protect against infections. However, despite these benefits, bifidobacteria still remain poorly understood at the biochemical, physiological and especially the genetic level. Herein we describe, for the first time, the development of a non-invasive luciferase-based reporter system for real-time tracking of Bifidobacterium species in vivo. Results The reporter vector pLuxMC1 is based on the recently described theta-type plasmid pBC1 from B. catenatulatum 1 and the luxABCDE operon from pPL2lux 2. Derivatives of pLuxMC1, harbouring a bifidobacterial promoter (pLuxMC2 as well as a synthetically derived promoter (pLuxMC3 3 placed upstream of luxABCDE, were constructed and found to stably replicate in B. breve UCC2003. The subsequent analysis of these strains allowed us to assess the functionality of pLuxMC1 both in vitro and in vivo. Conclusion Our results demonstrate the potential of pLuxMC1 as a real-time, non-invasive reporter system for Bifidobacterium. It has also allowed us, for the first time, to track the colonisation potential and persistence of this probiotic species in real time. An interesting and significant outcome of the study is the identification of the caecum as a niche environment for B. breve UCC2003 within the mouse gastrointestinal tract (GI tract.

  9. Aptamer-based viability impedimetric sensor for bacteria.

    Science.gov (United States)

    Labib, Mahmoud; Zamay, Anna S; Kolovskaya, Olga S; Reshetneva, Irina T; Zamay, Galina S; Kibbee, Richard J; Sattar, Syed A; Zamay, Tatiana N; Berezovski, Maxim V

    2012-11-06

    The development of an aptamer-based viability impedimetric sensor for bacteria (AptaVISens-B) is presented. Highly specific DNA aptamers to live Salmonella typhimurium were selected via the cell-systematic evolution of ligands by exponential enrichment (SELEX) technique. Twelve rounds of selection were performed; each comprises a positive selection step against viable S. typhimurium and a negative selection step against heat killed S. typhimurium and a mixture of related pathogens, including Salmonella enteritidis, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Citrobacter freundii to ensure the species specificity of the selected aptamers. The DNA sequence showing the highest binding affinity to the bacteria was further integrated into an impedimetric sensor via self-assembly onto a gold nanoparticle-modified screen-printed carbon electrode (GNP-SPCE). Remarkably, this aptasensor is highly selective and can successfully detect S. typhimurium down to 600 CFU mL(-1) (equivalent to 18 live cells in 30 μL of assay volume) and distinguish it from other Salmonella species, including S. enteritidis and S. choleraesuis. This report is envisaged to open a new venue for the aptamer-based viability sensing of a variety of microorganisms, particularly viable but nonculturable (VBNC) bacteria, using a rapid, economic, and label-free electrochemical platform.

  10. Use of nuclear receptor luciferase-based bioassays to detect endocrine active chemicals in a biosolids-biochar amended soil.

    Science.gov (United States)

    Anderson, Carolyn G; Joshi, Geetika; Bair, Daniel A; Oriol, Charlotte; He, Guochun; Parikh, Sanjai J; Denison, Michael S; Scow, Kate M

    2017-08-01

    Biosolids are a potentially valuable source of carbon and nutrients for agricultural soils; however, potential unintended impacts on human health and the environment must be considered. Virtually all biosolids contain trace amounts endocrine-disrupting chemicals derived from human use of pharmaceuticals and personal care products (PPCPs). One potential way to reduce the bioavailability of PPCPs is to co-apply biosolids with biochar to soil, because biochar's chemical (e.g., aromaticity) and physical properties (e.g., surface area) give it a high affinity to bind many organic chemicals in the environment. We developed a soil-specific extraction method and utilized a luciferase-based bioassay (CALUX) to detect endocrine active chemicals in a biosolids-biochar co-amendment soil greenhouse study. Both biochar (walnut shell, 900 °C) and biosolids had positive impacts on carrot and lettuce biomass accumulation over our study period. However, the walnut shell biochar stimulated aryl hydrocarbon receptor activity, suggesting the presence of potential endocrine active chemicals in the biochar. Since the biochar rate tested (100 t ha -1 ) is above the average agronomic rate (10-20 t ha -1 ), endocrine effects would not be expected in most environmental applications. The effect of high temperature biochars on endocrine system pathways must be explored further, using both quantitative analytical tools to identify potential endocrine active chemicals and highly sensitive bioanalytical assays such as CALUX to measure the resulting biological activity of such compounds. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Identification of luteolin as enterovirus 71 and coxsackievirus A16 inhibitors through reporter viruses and cell viability-based screening.

    Science.gov (United States)

    Xu, Lin; Su, Weiheng; Jin, Jun; Chen, Jiawen; Li, Xiaojun; Zhang, Xuyuan; Sun, Meiyan; Sun, Shiyang; Fan, Peihu; An, Dong; Zhang, Huafei; Zhang, Xiguang; Kong, Wei; Ma, Tonghui; Jiang, Chunlai

    2014-07-17

    Hand, foot and mouth disease (HFMD) is a common pediatric illness mainly caused by infection with enterovirus 71 (EV71) and coxsackievirus A16 (CA16). The frequent HFMD outbreaks have become a serious public health problem. Currently, no vaccine or antiviral drug for EV71/CA16 infections has been approved. In this study, a two-step screening platform consisting of reporter virus-based assays and cell viability‑based assays was developed to identify potential inhibitors of EV71/CA16 infection. Two types of reporter viruses, a pseudovirus containing luciferase-encoding RNA replicons encapsidated by viral capsid proteins and a full-length reporter virus containing enhanced green fluorescent protein, were used for primary screening of 400 highly purified natural compounds. Thereafter, a cell viability-based secondary screen was performed for the identified hits to confirm their antiviral activities. Three compounds (luteolin, galangin, and quercetin) were identified, among which luteolin exhibited the most potent inhibition of viral infection. In the cell viability assay and plaque reduction assay, luteolin showed similar 50% effective concentration (EC50) values of about 10 μM. Luteolin targeted the post-attachment stage of EV71 and CA16 infection by inhibiting viral RNA replication. This study suggests that luteolin may serve as a lead compound to develop potent anti-EV71 and CA16 drugs.

  12. Development of HSPA1A promoter-driven luciferase reporter gene assays in human cells for assessing the oxidative damage induced by silver nanoparticles

    International Nuclear Information System (INIS)

    Xin, Lili; Wang, Jianshu; Zhang, Leshuai W.; Che, Bizhong; Dong, Guangzhu; Fan, Guoqiang; Cheng, Kaiming

    2016-01-01

    The exponential increase in the total number of engineered nanoparticles in consumer products requires novel tools for rapid and cost-effective toxicology screening. In order to assess the oxidative damage induced by nanoparticles, toxicity test systems based on a human HSPA1A promoter-driven luciferase reporter in HepG2, LO2, A549, and HBE cells were established. After treated with heat shock and a group of silver nanoparticles (AgNPs) with different primary particle sizes, the cell viability, oxidative damage, and luciferase activity were determined. The time-dependent Ag + ions release from AgNPs in cell medium was also evaluated. Our results showed that heat shock produced a strong time-dependent induction of relative luciferase activity in the four luciferase reporter cells. Surprisingly, at 4 h of recovery, the relative luciferase activity was > 98 × the control level in HepG2-luciferase cells. Exposure to different sizes of AgNPs resulted in activation of the HSPA1A promoter in a dose-dependent manner, even at low cytotoxic or non-cytotoxic doses. The smaller (5 nm) AgNPs were more potent in luciferase induction than the larger (50 and 75 nm) AgNPs. These results were generally in accordance with the oxidative damage indicated by malondialdehyde concentration, reactive oxygen species induction and glutathione depletion, and Ag + ions release in cell medium. Compared with the other three luciferase reporter cells, the luciferase signal in HepG2-luciferase cells is obviously more sensitive and stable. We conclude that the luciferase reporter cells, especially the HepG2-luciferase cells, could provide a valuable tool for rapid screening of the oxidative damage induced by AgNPs. - Highlights: • We established the stable HSPA1A promoter-driven luciferase reporter cells. • Silver nanoparticles induced dose-dependent increases in luciferase activity. • HSPA1A promoter activity is a sensitive and responsive indicator of oxidative stress. • HepG2-luciferase

  13. Development of HSPA1A promoter-driven luciferase reporter gene assays in human cells for assessing the oxidative damage induced by silver nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Xin, Lili, E-mail: llxin@suda.edu.cn [School of Public Health, Medical College of Soochow University, 199 Renai Road, Suzhou 215123, Jiangsu (China); Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, School of Public Health, Soochow University, Suzhou 215123 (China); Wang, Jianshu [Suzhou Center for Disease Prevention and Control, 72 Sanxiang Road, Suzhou, Jiangsu (China); Zhang, Leshuai W. [School of Radiation Medicine and Protection & School for Radiological and Interdisciplinary Sciences (RAD-X), Soochow University, 215123 (China); Che, Bizhong; Dong, Guangzhu [School of Public Health, Medical College of Soochow University, 199 Renai Road, Suzhou 215123, Jiangsu (China); Fan, Guoqiang; Cheng, Kaiming [Suzhou Industrial Park Centers for Disease Control and Prevention, 58 Suqian Road, Suzhou, Jiangsu (China)

    2016-08-01

    The exponential increase in the total number of engineered nanoparticles in consumer products requires novel tools for rapid and cost-effective toxicology screening. In order to assess the oxidative damage induced by nanoparticles, toxicity test systems based on a human HSPA1A promoter-driven luciferase reporter in HepG2, LO2, A549, and HBE cells were established. After treated with heat shock and a group of silver nanoparticles (AgNPs) with different primary particle sizes, the cell viability, oxidative damage, and luciferase activity were determined. The time-dependent Ag{sup +} ions release from AgNPs in cell medium was also evaluated. Our results showed that heat shock produced a strong time-dependent induction of relative luciferase activity in the four luciferase reporter cells. Surprisingly, at 4 h of recovery, the relative luciferase activity was > 98 × the control level in HepG2-luciferase cells. Exposure to different sizes of AgNPs resulted in activation of the HSPA1A promoter in a dose-dependent manner, even at low cytotoxic or non-cytotoxic doses. The smaller (5 nm) AgNPs were more potent in luciferase induction than the larger (50 and 75 nm) AgNPs. These results were generally in accordance with the oxidative damage indicated by malondialdehyde concentration, reactive oxygen species induction and glutathione depletion, and Ag{sup +} ions release in cell medium. Compared with the other three luciferase reporter cells, the luciferase signal in HepG2-luciferase cells is obviously more sensitive and stable. We conclude that the luciferase reporter cells, especially the HepG2-luciferase cells, could provide a valuable tool for rapid screening of the oxidative damage induced by AgNPs. - Highlights: • We established the stable HSPA1A promoter-driven luciferase reporter cells. • Silver nanoparticles induced dose-dependent increases in luciferase activity. • HSPA1A promoter activity is a sensitive and responsive indicator of oxidative stress. • HepG2

  14. Bioluminescence-based cytotoxicity assay for simultaneous evaluation of cell viability and membrane damage in human hepatoma HepG2 cells.

    Science.gov (United States)

    Uno, Katsuhiro; Murotomi, Kazutoshi; Kazuki, Yasuhiro; Oshimura, Mitsuo; Nakajima, Yoshihiro

    2018-05-01

    We have developed a bioluminescence-based non-destructive cytotoxicity assay in which cell viability and membrane damage are simultaneously evaluated using Emerald luciferase (ELuc) and endoplasmic reticulum (ER)-targeted copepod luciferase (GLuc-KDEL), respectively, by using multi-integrase mouse artificial chromosome (MI-MAC) vector. We have demonstrated that the time-dependent concentration response curves of ELuc luminescence intensity and WST-1 assay, and GLuc-KDEL luminescence intensity and lactate dehydrogenase (LDH) activity in the culture medium accompanied by cytotoxicity show good agreement in toxicant-treated ELuc- and GLuc-KDEL-expressing HepG2 stable cell lines. We have clarified that the increase of GLuc-KDEL luminescence intensity in the culture medium reflects the type of cell death, including necrosis and late apoptosis, but not early apoptosis. We have also uncovered a strong correlation between GLuc-KDEL luminescence intensity in the culture medium and the extracellular release of high mobility group box 1 (HMGB1), a representative damage-associated molecular pattern (DAMP) molecule. The bioluminescence measurement assay using ELuc and GLuc-KDEL developed in this study can simultaneously monitor cell viability and membrane damage, respectively, and the increase of GLuc-KDEL luminescence intensity in the culture medium accompanied by the increase of cytotoxicity is an index of necrosis and late apoptosis associated with the extracellular release of DAMP molecules. Copyright © 2018 John Wiley & Sons, Ltd.

  15. Optimizing cell viability in droplet-based cell deposition

    NARCIS (Netherlands)

    Hendriks, Jan; Visser, C.W.; Henke, S.J.; Leijten, Jeroen Christianus Hermanus; Saris, Daniël B.F.; Sun, Chao; Lohse, Detlef; Karperien, Hermanus Bernardus Johannes

    2015-01-01

    Biofabrication commonly involves the use of liquid droplets to transport cells to the printed structure. However, the viability of the cells after impact is poorly controlled and understood, hampering applications including cell spraying, inkjet bioprinting, and laser-assisted cell transfer. Here,

  16. Development and validation of cell-based luciferase reporter gene assays for measuring neutralizing anti-drug antibodies against interferon beta

    DEFF Research Database (Denmark)

    Hermanrud, Christina; Ryner, Malin; Luft, Thomas

    2016-01-01

    a normal distribution for the majority of runs, allowing a parametric approach for cut-point calculation to be used, where NAb positive samples could be identified with 95% confidence. An analysis of means and variances indicated that a floating cut-point should be used for all assays. The assays......Neutralizing anti-drug antibodies (NAbs) against therapeutic interferon beta (IFNβ) in people with multiple sclerosis (MS) are measured with cell-based bioassays. The aim of this study was to redevelop and validate two luciferase reporter-gene bioassays, LUC and iLite, using a cut-point approach...... to identify NAb positive samples. Such an approach is favored by the pharmaceutical industry and governmental regulatory agencies as it has a clear statistical basis and overcomes the limitations of the current assays based on the Kawade principle. The work was conducted following the latest assay guidelines...

  17. Establishment of a luciferase assay-based screening system: Fumitremorgin C selectively inhibits cellular proliferation of immortalized astrocytes expressing an active form of AKT

    International Nuclear Information System (INIS)

    Wang Lei; Sasai, Ken; Akagi, Tsuyoshi; Tanaka, Shinya

    2008-01-01

    The AKT pathway is frequently activated in glioblastoma, and as such, inhibitors of this pathway could prove very useful as anti-glioblastoma therapies. Here we established immortalized astrocytes expressing Renilla luciferase as well as those expressing both an active form of AKT and firefly luciferase. Since both luciferase activities represent the numbers of corresponding cell lines, novel inhibitors of the AKT pathway can be identified by treating co-cultures containing the two types of luciferase-expressing cells with individual compounds. Indeed, such a screening system succeeded in identifying fumitremorgin C as an efficient inhibitor of the AKT pathway, which was further confirmed by the ability of fumitremorgin C to selectively inhibit the growth of immortalized astrocytes expressing an active form of AKT. The present study proposes a broadly applicable approach for identifying therapeutic agents that target the pathways and/or molecules responsible for cancer development

  18. Landscape-based population viability models demonstrate importance of strategic conservation planning for birds

    Science.gov (United States)

    Thomas W. Bonnot; Frank R. Thompson; Joshua J. Millspaugh; D. Todd. Jones-Farland

    2013-01-01

    Efforts to conserve regional biodiversity in the face of global climate change, habitat loss and fragmentation will depend on approaches that consider population processes at multiple scales. By combining habitat and demographic modeling, landscape-based population viability models effectively relate small-scale habitat and landscape patterns to regional population...

  19. Development and validation of cell-based luciferase reporter gene assays for measuring neutralizing anti-drug antibodies against interferon beta.

    Science.gov (United States)

    Hermanrud, Christina; Ryner, Malin; Luft, Thomas; Jensen, Poul Erik; Ingenhoven, Kathleen; Rat, Dorothea; Deisenhammer, Florian; Sørensen, Per Soelberg; Pallardy, Marc; Sikkema, Dan; Bertotti, Elisa; Kramer, Daniel; Creeke, Paul; Fogdell-Hahn, Anna

    2016-03-01

    Neutralizing anti-drug antibodies (NAbs) against therapeutic interferon beta (IFNβ) in people with multiple sclerosis (MS) are measured with cell-based bioassays. The aim of this study was to redevelop and validate two luciferase reporter-gene bioassays, LUC and iLite, using a cut-point approach to identify NAb positive samples. Such an approach is favored by the pharmaceutical industry and governmental regulatory agencies as it has a clear statistical basis and overcomes the limitations of the current assays based on the Kawade principle. The work was conducted following the latest assay guidelines. The assays were re-developed and validated as part of the "Anti-Biopharmaceutical Immunization: Prediction and analysis of clinical relevance to minimize the risk" (ABIRISK) consortium and involved a joint collaboration between four academic laboratories and two pharmaceutical companies. The LUC assay was validated at Innsbruck Medical University (LUCIMU) and at Rigshospitalet (LUCRH) Copenhagen, and the iLite assay at Karolinska Institutet, Stockholm. For both assays, the optimal serum sample concentration in relation to sensitivity and recovery was 2.5% (v/v) in assay media. A Shapiro-Wilk test indicated a normal distribution for the majority of runs, allowing a parametric approach for cut-point calculation to be used, where NAb positive samples could be identified with 95% confidence. An analysis of means and variances indicated that a floating cut-point should be used for all assays. The assays demonstrated acceptable sensitivity for being cell-based assays, with a confirmed limit of detection in neat serum of 1519 ng/mL for LUCIMU, 814 ng/mL for LUCRH, and 320 ng/mL for iLite. Use of the validated cut-point assay, in comparison with the previously used Kawade method, identified 14% more NAb positive samples. In conclusion, implementation of the cut-point design resulted in increased sensitivity to detect NAbs. However, the clinical significance of these low

  20. Sustainable model for financial viability of decentralized biomass gasifier based power projects

    International Nuclear Information System (INIS)

    Palit, Debajit; Malhotra, Ramit; Kumar, Atul

    2011-01-01

    This paper made a modest attempt for designing a sustainable model for financial viability of biomass gasifier power projects for enhancing electricity access in India and other developing countries. For long term sustainability of distributed generation projects in remote rural areas, viability from both project implementing agency (PIA) and the end-users need to be ensured. The minimum required prices of electricity from both PIA and end-user perspective have been estimated. While for PIA the cost recovery is the key for viability, the affordability to pay the electricity cost is crucial for the end users. Analysis carried out in this paper on the basis of data obtained from operational projects implemented in India reveal that it is essential to operate the system at a higher capacity utilization factor. While this can be achieved though creating convergence with locally relevant economic activity, it is also observed that micro-enterprises cannot pay beyond a certain price of electricity to keep it sustainable. This paper sets forth a case for developing a regulatory mechanism to extend the tariff fixation for the projects and providing cross-subsidies to ensure long term sustainability of off-grid project. - Highlights: → We design sustainable financial model for viability of biomass gasifier projects. → Analysis based on field data obtained from operational projects in India. Estimated electricity pricing from both implementing agency and end-users perspective. → A regulatory mechanism for tariff fixation and cross subsidization is recommended.

  1. Relationship of the luminous bacterial symbiont of the Caribbean flashlight fish, Kryptophanaron alfredi (family Anomalopidae) to other luminous bacteria based on bacterial luciferase (luxA) genes.

    Science.gov (United States)

    Haygood, M G

    1990-01-01

    Flashlight fishes (family Anomalopidae) have light organs that contain luminous bacterial symbionts. Although the symbionts have not yet been successfully cultured, the luciferase genes have been cloned directly from the light organ of the Caribbean species, Kryptophanaron alfredi. The goal of this project was to evaluate the relationship of the symbiont to free-living luminous bacteria by comparison of genes coding for bacterial luciferase (lux genes). Hybridization of a lux AB probe from the Kryptophanaron alfredi symbiont to DNAs from 9 strains (8 species) of luminous bacteria showed that none of the strains tested had lux genes highly similar to the symbiont. The most similar were a group consisting of Vibrio harveyi, Vibrio splendidus and Vibrio orientalis. The nucleotide sequence of the luciferase alpha subunit gene luxA) of the Kryptophanaron alfredi symbiont was determined in order to do a more detailed comparison with published luxA sequences from Vibrio harveyi, Vibrio fischeri and Photobacterium leiognathi. The hybridization results, sequence comparisons and the mol% G + C of the Kryptophanaron alfredi symbiont luxA gene suggest that the symbiont may be considered as a new species of luminous Vibrio related to Vibrio harveyi.

  2. More Pitfalls with Sperm Viability Staining and a Viability-Based Stress Test to Characterize Sperm Quality

    Directory of Open Access Journals (Sweden)

    Barbara A. Eckel

    2017-12-01

    Full Text Available Sperm viability (SV, the proportion of live sperm in a sample, is a widely applied measure of sperm quality but few studies test its robustness. At least three reasons make SV problematic as a surrogate for sperm quality. First, reviewing the ecological literature revealed that previously identified methodological pitfalls have not been overcome, including low cross-study standardization of protocols, inadequate statistical treatment, and unaccounted for within-sample heterogeneity. Second, SV is affected by biological variation such as between species, reproductive organs, or sperm age cohorts. Third, the proportion of live sperm extracted from males appears more related to male than to sperm quality in the sense of the future performance of sperm. We propose an alternative method to assess sperm quality by characterizing the temporal decrease of SV in a stressor medium and illustrate in two species, the common bedbug (Cimex lectularius and the fruit fly (Drosophila melanogaster how some common methodological pitfalls may be circumvented. Our data empirically support the well-known but little-considered facts that (i non-blind measurements may alter SV and (ii that SV frequently have non-significant repeatability within one sample. (iii Cross-sectional sampling of ejaculates showed that this heterogeneity even masked a biological pattern—the sperm stratification within males. We show (iv that this shortcoming can be overcome by following the temporal decline of SV of a sperm subsample in a stress test. Finally, (v comparing the staining pattern of sperm between Cimex and Drosophila, we found that in the latter, the visibility of sperm is substantially delayed (30 min when sperm density is high. We show that this delay in stained sperm visibility was, however, not biased toward dead or live sperm. To measure sperm quality, we advocate analyzing the temporal decline in SV in a stressor medium over current protocols that use SV per se and

  3. Tibetan Firefly Luciferase with Low Temperature Adaptation.

    Science.gov (United States)

    Mitani, Yasuo; Futahashi, Ryo; Liu, Zichao; Liang, Xingcai; Ohmiya, Yoshihiro

    2017-03-01

    Fireflies are widespread all over the world and a numerous numbers of luciferases have been isolated and characterized. In this study, we identified and characterized the luciferase and luciferase-like genes from a Tibetan firefly collected in Shangri-La, China. The altitude of this area is more than 3300 m. We saw this Tibetan firefly flying with strong luminescence after sunset at ~10°C. We analyzed the transcriptome of Tibetan firefly using head, thorax, abdomen (without light organ), and light organ tissue by RNA sequencing. We identified one luciferase gene, which was almost identical to luciferase from fireflies Pyrocoelia species, and expressed specifically in the light organ. Interestingly, the optimal temperature of the Tibetan firefly recombinant luciferase was 10°C. The K m for D-luciferin and ATP of the recombinant luciferase was 23 and 154 μm, respectively. The optimal pH was around 7.0-7.5. The emission peak was 556 nm at pH 8.0, while it shifted to 606 nm at pH 6.0. We also found a luciferase-like gene with 43% identical amino acids to the Tibetan firefly luciferase, which was scarcely expressed in any portion of the adult body. No luciferase activity was detected for this luciferase-like protein. © 2016 The American Society of Photobiology.

  4. Assessing Viability and Sustainability: a Systems-based Approach for Deriving Comprehensive Indicator Sets

    Directory of Open Access Journals (Sweden)

    Hartmut Bossel

    2002-01-01

    Full Text Available Performance assessment in holistic approaches such as integrated natural resource management has to deal with a complex set of interacting and self-organizing natural and human systems and agents, all pursuing their own "interests" while also contributing to the development of the total system. Performance indicators must therefore reflect the viability of essential component systems as well as their contributions to the viability and performance of other component systems and the total system under study. A systems-based derivation of a comprehensive set of performance indicators first requires the identification of essential component systems, their mutual (often hierarchical or reciprocal relationships, and their contributions to the performance of other component systems and the total system. The second step consists of identifying the indicators that represent the viability states of the component systems and the contributions of these component systems to the performance of the total system. The search for performance indicators is guided by the realization that essential interests (orientations or orientors of systems and actors are shaped by both their characteristic functions and the fundamental and general properties of their system environments (e.g., normal environmental state, scarcity of resources, variety, variability, change, other coexisting systems. To be viable, a system must devote an essential minimum amount of attention to satisfying the "basic orientors" that respond to the properties of its environment. This fact can be used to define comprehensive and system-specific sets of performance indicators that reflect all important concerns. Often, qualitative indicators and the study of qualitative systems are sufficient for reliable performance assessments. However, this approach can also be formalized for quantitative computer-assisted assessment. Examples are presented of indicator sets for the sustainable development of

  5. Tissues viability and blood flow sensing based on a new nanophotonics method

    Science.gov (United States)

    Yariv, Inbar; Haddad, Menashe; Duadi, Hamootal; Motiei, Menachem; Fixler, Dror

    2018-02-01

    Extracting optical parameters of turbid medium (e.g. tissue) by light reflectance signals is of great interest and has many applications in the medical world, life science, material analysis and biomedical optics. The reemitted light from an irradiated tissue is affected by the light's interaction with the tissue components and contains the information about the tissue structure and physiological state. In this research we present a novel noninvasive nanophotonics technique, i.e., iterative multi-plane optical property extraction (IMOPE) based on reflectance measurements. The reflectance based IMOPE was applied for tissue viability examination, detection of gold nanorods (GNRs) within the blood circulation as well as blood flow detection using the GNRs presence within the blood vessels. The basics of the IMOPE combine a simple experimental setup for recording light intensity images with an iterative Gerchberg-Saxton (G-S) algorithm for reconstructing the reflected light phase and computing its standard deviation (STD). Changes in tissue composition affect its optical properties which results in changes in the light phase that can be measured by its STD. This work presents reflectance based IMOPE tissue viability examination, producing a decrease in the computed STD for older tissues, as well as investigating their organic material absorption capability. Finally, differentiation of the femoral vein from adjacent tissues using GNRs and the detection of their presence within blood circulation and tissues are also presented with high sensitivity (better than computed tomography) to low quantities of GNRs (<3 mg).

  6. An individual-based model for population viability analysis of humpback chub in Grand Canyon

    Science.gov (United States)

    Pine, William Pine; Healy, Brian; Smith, Emily Omana; Trammell, Melissa; Speas, Dave; Valdez, Rich; Yard, Mike; Walters, Carl; Ahrens, Rob; Vanhaverbeke, Randy; Stone, Dennis; Wilson, Wade

    2013-01-01

    We developed an individual-based population viability analysis model (females only) for evaluating risk to populations from catastrophic events or conservation and research actions. This model tracks attributes (size, weight, viability, etc.) for individual fish through time and then compiles this information to assess the extinction risk of the population across large numbers of simulation trials. Using a case history for the Little Colorado River population of Humpback Chub Gila cypha in Grand Canyon, Arizona, we assessed extinction risk and resiliency to a catastrophic event for this population and then assessed a series of conservation actions related to removing specific numbers of Humpback Chub at different sizes for conservation purposes, such as translocating individuals to establish other spawning populations or hatchery refuge development. Our results suggested that the Little Colorado River population is generally resilient to a single catastrophic event and also to removals of larvae and juveniles for conservation purposes, including translocations to establish new populations. Our results also suggested that translocation success is dependent on similar survival rates in receiving and donor streams and low emigration rates from recipient streams. In addition, translocating either large numbers of larvae or small numbers of large juveniles has generally an equal likelihood of successful population establishment at similar extinction risk levels to the Little Colorado River donor population. Our model created a transparent platform to consider extinction risk to populations from catastrophe or conservation actions and should prove useful to managers assessing these risks for endangered species such as Humpback Chub.

  7. Study of functional viability of SU-8-based microneedles for neural applications

    International Nuclear Information System (INIS)

    Fernández, Luis J; Altuna, Ane; Tijero, Maria; Vilares, Roman; Berganzo, Javier; Blanco, F J; Gabriel, Gemma; Villa, Rosa; Rodríguez, Manuel J; Batlle, Montse

    2009-01-01

    This paper presents the design, fabrication, packaging and first test results of SU-8-based microneedles for neural applications. By the use of photolithography, sputtering and bonding techniques, polymer needles with integrated microchannels and electrodes have been successfully fabricated. The use of photolithography for the patterning of the fluidic channel integrated in the needle allows the design of multiple outlet ports at the needle tip, minimizing the possibility of being blocked by the tissue. Furthermore, the flexibility of the polymer reduces the risk of fracture and tissue damage once the needle is inserted, while it is still rigid enough to allow a perfect insertion into the neural tissue. Fluidic and electric characterization of the microneedles has shown their viability for drug delivery and monitoring in neural applications. First drug delivery tests in ex vivo tissue demonstrated the functional viability of the needle to deliver drugs to precise points. Furthermore, in vivo experiments have demonstrated lower associated damages during insertion than those by stereotaxic standard needles

  8. Viability Theory

    CERN Document Server

    Aubin, Jean-Pierre; Saint-Pierre, Patrick

    2011-01-01

    Viability theory designs and develops mathematical and algorithmic methods for investigating the adaptation to viability constraints of evolutions governed by complex systems under uncertainty that are found in many domains involving living beings, from biological evolution to economics, from environmental sciences to financial markets, from control theory and robotics to cognitive sciences. It involves interdisciplinary investigations spanning fields that have traditionally developed in isolation. The purpose of this book is to present an initiation to applications of viability theory, explai

  9. Sustainable model for financial viability of decentralized biomass gasifier based power projects

    NARCIS (Netherlands)

    Palit, D.; Malhotra, R.; Kumar, Atul

    2011-01-01

    This paper made a modest attempt for designing a sustainable model for financial viability of biomass gasifier power projects for enhancing electricity access in India and other developing countries. For long term sustainability of distributed generation projects in remote rural areas, viability

  10. Improvement of thermostability and activity of firefly luciferase through [TMG][Ac] ionic liquid mediator.

    Science.gov (United States)

    Ebrahimi, Mehdi; Hosseinkhani, Saman; Heydari, Akbar; Khavari-Nejad, Ramazan Ali; Akbari, Jafar

    2012-10-01

    Firefly luciferase catalyzes production of light from luciferin in the presence of Mg(2+)-ATP and oxygen. This enzyme has wide range of applications in biotechnology and development of biosensors. The low thermal stability of wild-type firefly luciferase is a limiting factor in most applications. Improvements in activity and stability of few enzymes in the presence of ionic liquids were shown in many reports. In this study, kinetic and thermal stability of firefly luciferase from Photinus pyralis in the presence of three tetramethylguanidine-based ionic liquids was investigated. The enzyme has shown improved activity in the presence of [1, 1, 3, 3-tetramethylguanidine][acetate], but in the presence of [TMG][trichloroacetate] and [TMG][triflouroacetate] activity, it decreased or unchanged significantly. Among these ionic liquids, only [TMG][Ac] has increased the thermal stability of luciferase. Incubation of [TMG][Ac] with firefly luciferase brought about with decrease of K(m) for ATP.

  11. Luciferase assay to study the activity of a cloned promoter DNA fragment.

    Science.gov (United States)

    Solberg, Nina; Krauss, Stefan

    2013-01-01

    Luciferase based assays have become an invaluable tool for the analysis of cloned promoter DNA fragments, both for verifying the ability of a potential promoter fragment to drive the expression of a luciferase reporter gene in various cellular contexts, and for dissecting binding elements in the promoter. Here, we describe the use of the Dual-Luciferase(®) Reporter Assay System created by Promega (Promega Corporation, Wisconsin, USA) to study the cloned 6.7 kilobases (kb) mouse (m) Tcf3 promoter DNA fragment in mouse embryonic derived neural stem cells (NSC). In this system, the expression of the firefly luciferase driven by the cloned mTcf3 promoter DNA fragment (including transcription initiation sites) is correlated with a co-transfected control reporter expressing Renilla luciferase from the herpes simplex virus (HSV) thymidine kinase promoter. Using an internal control reporter allows to normalize the activity of the experimental reporter to the internal control, which minimizes experimental variability.

  12. The role of taxation policy and incentives in wind-based distributed generation projects viability. Ontario case study

    International Nuclear Information System (INIS)

    Albadi, M.H.; El-Saadany, E.F.

    2009-01-01

    Taxation policy and incentives play a vital role in wind-based distributed generation projects viability. In this paper, a thorough techno-economical evaluation of wind-based distributed generation projects is conducted to investigate the effect of taxes and incentives in the economic viability of investments in this sector. This paper considers the effects of Provincial income taxes, capital cost allowance (CCA), property taxes, and wind power production Federal incentives. The case study is conducted for different wind turbines and wind speed scenarios. Given turbine and wind speed data, the Capacity Factor (CF) of each turbine and wind speed scenario was calculated. Net Present Value (NPV) and Internal Rate of Return (IRR) for different scenarios were then used to assess the project's viability considering Ontario Standard Offer Program (SOP) for wind power. (author)

  13. Nanoscale size effect in in situ titanium based composites with cell viability and cytocompatibility studies

    Energy Technology Data Exchange (ETDEWEB)

    Miklaszewski, Andrzej, E-mail: andrzej.miklaszewski@put.poznan.pl [Institute of Materials Science and Engineering, Poznan University of Technology, Jana Pawla II 24, 61-138 Poznan (Poland); Jurczyk, Mieczysława U. [Division Mother' s and Child' s Health, Poznan University of Medical Sciences, Polna 33, 60-535 Poznan (Poland); Kaczmarek, Mariusz [Department of Immunology, Chair of Clinical Immunology, Poznan University of Medical Sciences, Rokietnicka 5D, 60-806 Poznan (Poland); Paszel-Jaworska, Anna; Romaniuk, Aleksandra; Lipińska, Natalia [Department of Clinical Chemistry and Molecular Diagnostics, Poznan University of Medical Sciences, Przybyszewskiego 49, 60-355 Poznan (Poland); Żurawski, Jakub [Department of Immunobiochemistry, Chair of Biology and Environmental Sciences, Poznan University of Medical Sciences, Rokietnicka 8, 60-806 Poznan (Poland); Urbaniak, Paulina [Department of Cell Biology, Poznan University of Medical Sciences, Rokietnicka 5D, 60-806 Poznan (Poland); Jurczyk, Mieczyslaw [Institute of Materials Science and Engineering, Poznan University of Technology, Jana Pawla II 24, 61-138 Poznan (Poland)

    2017-04-01

    Novel in situ Metal Matrix Nanocomposite (MMNC) materials based on titanium and boron, revealed their new properties in the nanoscale range. In situ nanocomposites, obtained through mechanical alloying and traditional powder metallurgy compaction and sintering, show obvious differences to their microstructural analogue. A unique microstructure connected with good mechanical properties reliant on the processing conditions favour the nanoscale range of results of the Ti-TiB in situ MMNC example. The data summarised in this work, support and extend the knowledge boundaries of the nanoscale size effect that influence not only the mechanical properties but also the studies on the cell viability and cytocompatibility. Prepared in the same bulk, in situ MMNC, based on titanium and boron, could be considered as a possible candidate for dental implants and other medical applications. The observed relations and research conclusions are transferable to the in situ MMNC material group. Aside from all the discussed relations, the increasing share of these composites in the ever-growing material markets, heavily depends on the attractiveness and a possible wider application of these composites as well as their operational simplicity presented in this work. - Highlights: • Nano and microscale size precursor influence the final composite microstructure and properties. • Obtained from the nanoscale precursor sinters, characterise with a uniform and highly dispersed microstructure • Mechanical properties favoured Nano scale size precursor • Boron addition could be significantly reduced for moderate properties range. • A possible candidate for dental implants and other medical applications.

  14. Morphological observation and analysis using automated image cytometry for the comparison of trypan blue and fluorescence-based viability detection method.

    Science.gov (United States)

    Chan, Leo Li-Ying; Kuksin, Dmitry; Laverty, Daniel J; Saldi, Stephanie; Qiu, Jean

    2015-05-01

    The ability to accurately determine cell viability is essential to performing a well-controlled biological experiment. Typical experiments range from standard cell culturing to advanced cell-based assays that may require cell viability measurement for downstream experiments. The traditional cell viability measurement method has been the trypan blue (TB) exclusion assay. However, since the introduction of fluorescence-based dyes for cell viability measurement using flow or image-based cytometry systems, there have been numerous publications comparing the two detection methods. Although previous studies have shown discrepancies between TB exclusion and fluorescence-based viability measurements, image-based morphological analysis was not performed in order to examine the viability discrepancies. In this work, we compared TB exclusion and fluorescence-based viability detection methods using image cytometry to observe morphological changes due to the effect of TB on dead cells. Imaging results showed that as the viability of a naturally-dying Jurkat cell sample decreased below 70 %, many TB-stained cells began to exhibit non-uniform morphological characteristics. Dead cells with these characteristics may be difficult to count under light microscopy, thus generating an artificially higher viability measurement compared to fluorescence-based method. These morphological observations can potentially explain the differences in viability measurement between the two methods.

  15. Experimental Calcium Silicate-Based Cement with and without Zirconium Oxide Modulates Fibroblasts Viability.

    Science.gov (United States)

    Slompo, Camila; Peres-Buzalaf, Camila; Gasque, Kellen Cristina da Silva; Damante, Carla Andreotti; Ordinola-Zapata, Ronald; Duarte, Marco Antonio Hungaro; de Oliveira, Rodrigo Cardoso

    2015-01-01

    The aim of this study was to verify whether the use of zirconium oxide as a radiopacifier of an experimental calcium silicate-based cement (WPCZO) leads to cytotoxicity. Fibroblasts were treated with different concentrations (10 mg/mL, 1 mg/mL, and 0.1 mg/mL) of the cements diluted in Dulbecco's modified Eagle's medium (DMEM) for periods of 12, 24, and 48 h. Groups tested were white Portland cement (WPC), white Portland cement with zirconium oxide (WPCZO), and white mineral trioxide aggregate Angelus (MTA). Control group cells were not treated. The cytotoxicity was evaluated through mitochondrial-activity (MTT) and cell-density (crystal violet) assays. All cements showed low cytotoxicity. In general, at the concentration of 10 mg/mL there was an increase in viability of those groups treated with WPC and WPCZO when compared to the control group (pcement with 20% zirconium oxide as the radiopacifier showed low cytotoxicity as a promising material to be exploited for root-end filling.

  16. On the Viability of Diffusion MRI-Based Microstructural Biomarkers in Ischemic Stroke

    Directory of Open Access Journals (Sweden)

    Ilaria Boscolo Galazzo

    2018-02-01

    Full Text Available Recent tract-based analyses provided evidence for the exploitability of 3D-SHORE microstructural descriptors derived from diffusion MRI (dMRI in revealing white matter (WM plasticity. In this work, we focused on the main open issues left: (1 the comparative analysis with respect to classical tensor-derived indices, i.e., Fractional Anisotropy (FA and Mean Diffusivity (MD; and (2 the ability to detect plasticity processes in gray matter (GM. Although signal modeling in GM is still largely unexplored, we investigated their sensibility to stroke-induced microstructural modifications occurring in the contralateral hemisphere. A more complete picture could provide hints for investigating the interplay of GM and WM modulations. Ten stroke patients and ten age/gender-matched healthy controls were enrolled in the study and underwent diffusion spectrum imaging (DSI. Acquisitions at three and two time points (tp were performed on patients and controls, respectively. For all subjects and acquisitions, FA and MD were computed along with 3D-SHORE-based indices [Generalized Fractional Anisotropy (GFA, Propagator Anisotropy (PA, Return To the Axis Probability (RTAP, Return To the Plane Probability (RTPP, and Mean Square Displacement (MSD]. Tract-based analysis involving the cortical, subcortical and transcallosal motor networks and region-based analysis in GM were successively performed, focusing on the contralateral hemisphere to the stroke. Reproducibility of all the indices on both WM and GM was quantitatively proved on controls. For tract-based, longitudinal group analyses revealed the highest significant differences across the subcortical and transcallosal networks for all the indices. The optimal regression model for predicting the clinical motor outcome at tp3 included GFA, PA, RTPP, and MSD in the subcortical network in combination with the main clinical information at baseline. Region-based analysis in the contralateral GM highlighted the ability of

  17. Fe65 does not stabilize AICD during activation of transcription in a luciferase assay

    International Nuclear Information System (INIS)

    Huysseune, Sandra; Kienlen-Campard, Pascal; Octave, Jean-Noel

    2007-01-01

    The APP intracellular domain (AICD) could be involved in signaling via interaction with the adaptor protein Fe65, and with the histone acetyl transferase Tip60. However, the real function of AICD and Fe65 in regulation of transcription remains controversial. In this study, the human APPGal4 fusion protein was expressed in CHO cells and the transcriptional activity of AICDGal4 was measured in a luciferase-based reporter assay. AICDGal4 was stabilized by expression of Fe65 and levels of AICDGal4 controlled luciferase activity. On the contrary, when human APP was expressed in CHO cells, coexpression of Fe65 increased luciferase activity without affecting the amount of AICD fragment. AICD produced from APP was protected from degradation by orthophenanthroline, but not by lactacystine, indicating that AICD is not a substrate of the chymotryptic activity of the proteasome. It is concluded that Fe65 can control luciferase activity without stabilizing the labile AICD fragment

  18. Compositional and physicochemical factors governing the viability of Lactobacillus rhamnosus GG embedded in starch-protein based edible films

    Science.gov (United States)

    Soukoulis, Christos; Singh, Poonam; Macnaughtan, William; Parmenter, Christopher; Fisk, Ian D.

    2016-01-01

    Probiotic incorporation in edible films and coatings has been shown recently to be an efficient strategy for the delivery of probiotics in foods. In the present work, the impact of the compositional, physicochemical and structural properties of binary starch-protein edible films on Lactobacillus rhamnosus GG viability and stability was evaluated. Native rice and corn starch, as well as bovine skin gelatine, sodium caseinate and soy protein concentrate were used for the fabrication of the probiotic edible films. Starch and protein type both impacted the structural, mechanical, optical and thermal properties of the films, and the process loss of L. rhamnosus GG during evaporation-dehydration was significantly lower in the presence of proteins (0.91–1.07 log CFU/g) compared to solely starch based systems (1.71 log CFU/g). A synergistic action between rice starch and proteins was detected when monitoring the viability of L. rhamnosus GG over four weeks at fridge and room temperature conditions. In particular, a 3- to 7-fold increase in the viability of L. rhamnosus GG was observed in the presence of proteins, with sodium caseinate – rice starch based films offering the most enhanced stability. The film's shelf-life (as calculated using the FAO/WHO (2011) basis of 6 log viable CFU/g) ranged between 27-96 and 15–24 days for systems stored at fridge or room temperature conditions respectively. PMID:26726280

  19. Firefly luciferase gene contains a cryptic promoter

    Czech Academy of Sciences Publication Activity Database

    Vopálenský, V.; Mašek, T.; Horváth, Ondřej; Vicenová, B.; Mokrejš, M.; Pospíšek, M.

    2008-01-01

    Roč. 14, č. 9 (2008), s. 1720-1729 ISSN 1355-8382 Grant - others:GAČR(CZ) GA204/03/1487; GAČR(CZ) GA301/07/0607; Mšk(CZ) LC06066 Program:LC Institutional research plan: CEZ:AV0Z50520514 Keywords : luciferase * cryptic promoter * hepatitis C virus Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.018, year: 2008

  20. Polyvinylpyrrolidone-Based Bio-Ink Improves Cell Viability and Homogeneity during Drop-On-Demand Printing

    Directory of Open Access Journals (Sweden)

    Wei Long Ng

    2017-02-01

    Full Text Available Drop-on-demand (DOD bioprinting has attracted huge attention for numerous biological applications due to its precise control over material volume and deposition pattern in a contactless printing approach. 3D bioprinting is still an emerging field and more work is required to improve the viability and homogeneity of printed cells during the printing process. Here, a general purpose bio-ink was developed using polyvinylpyrrolidone (PVP macromolecules. Different PVP-based bio-inks (0%–3% w/v were prepared and evaluated for their printability; the short-term and long-term viability of the printed cells were first investigated. The Z value of a bio-ink determines its printability; it is the inverse of the Ohnesorge number (Oh, which is the ratio between the Reynolds number and a square root of the Weber number, and is independent of the bio-ink velocity. The viability of printed cells is dependent on the Z values of the bio-inks; the results indicated that the cells can be printed without any significant impairment using a bio-ink with a threshold Z value of ≤9.30 (2% and 2.5% w/v. Next, the cell output was evaluated over a period of 30 min. The results indicated that PVP molecules mitigate the cell adhesion and sedimentation during the printing process; the 2.5% w/v PVP bio-ink demonstrated the most consistent cell output over a period of 30 min. Hence, PVP macromolecules can play a critical role in improving the cell viability and homogeneity during the bioprinting process.

  1. Evaluating the potential for weed seed dispersal based on waterfowl consumption and seed viability.

    Science.gov (United States)

    Farmer, Jaime A; Webb, Elisabeth B; Pierce, Robert A; Bradley, Kevin W

    2017-12-01

    Migratory waterfowl have often been implicated in the movement of troublesome agronomic and wetland weed species. However, minimal research has been conducted to investigate the dispersal of agronomically important weed species by waterfowl. The two objectives for this project were to determine what weed species are being consumed by ducks and snow geese, and to determine the recovery rate and viability of 13 agronomic weed species after passage through a duck's digestive system. Seed recovered from digestive tracts of 526 ducks and geese harvested during a 2-year field study had 35 020 plants emerge. A greater variety of plant species emerged from ducks each year (47 and 31 species) compared to geese (11 and 3 species). Viable seed from 11 of 13 weed species fed to ducks in a controlled feeding study were recovered. Viability rate and gut retention times indicated potential dispersal up to 2900 km from the source depending on seed characteristics and variability in waterfowl dispersal distances. Study results confirm that waterfowl are consuming seeds from a variety of agronomically important weed species, including Palmer amaranth, which can remain viable after passage through digestive tracts and have potential to be dispersed over long distances by waterfowl. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  2. Re-engineering of Bacterial Luciferase; For New Aspects of Bioluminescence.

    Science.gov (United States)

    Kim, Da-Som; Choi, Jeong-Ran; Ko, Jeong-Ae; Kim, Kangmin

    2018-01-01

    Bacterial luminescence is the end-product of biochemical reactions catalyzed by the luciferase enzyme. Nowadays, this fascinating phenomenon has been widely used as reporter and/or sensors to detect a variety of biological and environmental processes. The enhancement or diversification of the luciferase activities will increase the versatility of bacterial luminescence. Here, to establish the strategy for luciferase engineering, we summarized the identity and relevant roles of key amino acid residues modulating luciferase in Vibrio harveyi, a model luminous bacterium. The current opinions on crystal structures and the critical amino acid residues involved in the substrate binding sites and unstructured loop have been delineated. Based on these, the potential target residues and/or parameters for enzyme engineering were also suggested in limited scale. In conclusion, even though the accurate knowledge on the bacterial luciferase is yet to be reported, the structure-guided site-directed mutagenesis approaches targeting the regulatory amino acids will provide a useful platform to re-engineer the bacterial luciferase in the future. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  3. Single cell adhesion force measurement for cell viability identification using an AFM cantilever-based micro putter

    Science.gov (United States)

    Shen, Yajing; Nakajima, Masahiro; Kojima, Seiji; Homma, Michio; Kojima, Masaru; Fukuda, Toshio

    2011-11-01

    Fast and sensitive cell viability identification is a key point for single cell analysis. To address this issue, this paper reports a novel single cell viability identification method based on the measurement of single cell shear adhesion force using an atomic force microscopy (AFM) cantilever-based micro putter. Viable and nonviable yeast cells are prepared and put onto three kinds of substrate surfaces, i.e. tungsten probe, gold and ITO substrate surfaces. A micro putter is fabricated from the AFM cantilever by focused ion beam etching technique. The spring constant of the micro putter is calibrated using the nanomanipulation approach. The shear adhesion force between the single viable or nonviable cell and each substrate is measured using the micro putter based on the nanorobotic manipulation system inside an environmental scanning electron microscope. The adhesion force is calculated based on the deflection of the micro putter beam. The results show that the adhesion force of the viable cell to the substrate is much larger than that of the nonviable cell. This identification method is label free, fast, sensitive and can give quantitative results at the single cell level.

  4. Development and validation of a quantitative, high-throughput, fluorescent-based bioassay to detect schistosoma viability.

    Directory of Open Access Journals (Sweden)

    Emily Peak

    2010-07-01

    Full Text Available Schistosomiasis, caused by infection with the blood fluke Schistosoma, is responsible for greater than 200,000 human deaths per annum. Objective high-throughput screens for detecting novel anti-schistosomal targets will drive 'genome to drug' lead translational science at an unprecedented rate. Current methods for detecting schistosome viability rely on qualitative microscopic criteria, which require an understanding of parasite morphology, and most importantly, must be subjectively interpreted. These limitations, in the current state of the art, have significantly impeded progress into whole schistosome screening for next generation chemotherapies.We present here a microtiter plate-based method for reproducibly detecting schistosomula viability that takes advantage of the differential uptake of fluorophores (propidium iodide and fluorescein diacetate by living organisms. We validate this high-throughput system in detecting schistosomula viability using auranofin (a known inhibitor of thioredoxin glutathione reductase, praziquantel and a range of small compounds with previously-described (gambogic acid, sodium salinomycin, ethinyl estradiol, fluoxetidine hydrochloride, miconazole nitrate, chlorpromazine hydrochloride, amphotericin b, niclosamide or suggested (bepridil, ciclopirox, rescinnamine, flucytosine, vinblastine and carbidopa anti-schistosomal activities. This developed method is sensitive (200 schistosomula/well can be assayed, relevant to industrial (384-well microtiter plate compatibility and academic (96-well microtiter plate compatibility settings, translatable to functional genomics screens and drug assays, does not require a priori knowledge of schistosome biology and is quantitative.The wide-scale application of this fluorescence-based bioassay will greatly accelerate the objective identification of novel therapeutic lead targets/compounds to combat schistosomiasis. Adapting this bioassay for use with other parasitic worm species

  5. New optical sensing technique of tissue viability and blood flow based on nanophotonic iterative multi-plane reflectance measurements

    Directory of Open Access Journals (Sweden)

    Yariv I

    2016-10-01

    Full Text Available Inbar Yariv,1 Menashe Haddad,2,3 Hamootal Duadi,1 Menachem Motiei,1 Dror Fixler1 1Faculty of Engineering and the Institute of Nanotechnology and Advanced Materials, Bar Ilan University, Ramat Gan, Israel; 2Sackler School of Medicine, Tel-Aviv University, Tel-Aviv, Israel; 3Mayanei Hayeshua Medical Center, Benei Brak, Israel Abstract: Physiological substances pose a challenge for researchers since their optical properties change constantly according to their physiological state. Examination of those substances noninvasively can be achieved by different optical methods with high sensitivity. Our research suggests the application of a novel noninvasive nanophotonics technique, ie, iterative multi-plane optical property extraction (IMOPE based on reflectance measurements, for tissue viability examination and gold nanorods (GNRs and blood flow detection. The IMOPE model combines an experimental setup designed for recording light intensity images with the multi-plane iterative Gerchberg-Saxton algorithm for reconstructing the reemitted light phase and calculating its standard deviation (STD. Changes in tissue composition affect its optical properties which results in changes in the light phase that can be measured by its STD. We have demonstrated this new concept of correlating the light phase STD and the optical properties of a substance, using transmission measurements only. This paper presents, for the first time, reflectance based IMOPE tissue viability examination, producing a decrease in the computed STD for older tissues, as well as investigating their organic material absorption capability. Finally, differentiation of the femoral vein from adjacent tissues using GNRs and the detection of their presence within blood circulation and tissues are also presented with high sensitivity (better than computed tomography to low quantities of GNRs (<3 mg. Keywords: Gerchberg-Saxton, optical properties, gold nanorods, blood vessel, tissue viability

  6. Viability and fertility of cooled equine semen diluted with skimmed milk or glycine egg yolk-based extenders

    Directory of Open Access Journals (Sweden)

    Guilherme Pugliesi

    2012-12-01

    Full Text Available Two semen extenders were compared for their ability to maintain viability of horse semen during 24 hours of cold preservation, and for the pregnancy rate after artificial insemination. In the experiment 1, five ejaculates from three stallions were split-diluted in either a skimmed milk-based extender (Kenney extender or a glycine egg yolk-based extender (Foote extender and cooled at 6-8 ºC for 24 hours. Semen samples stored in Kenney extender for 24 hours had higher motility and spermatic vigor compared with those stored in Foote extender. However, samples stored in Foote extender had higher number of reactive sperm by hypoosmotic test and greater viability by epifluorescence test compared with those in Kenney extender. In the experiment 2, 17 and 23 ejaculates from two stallions were split-diluted with Kenney extender and Foote extender. The sperm concentration in each extender was adjusted to 500 million viable sperms per insemination dose. Semen was cooled to 6-8 ºC and stored for 24 hours. Seventy-four cycles of crossbred mares were inseminated with either semen diluted in Kenney extender or semen diluted in Foote extender. The pregnancy rate was higher from semen diluted in Kenney extender than that from semen in Foote extender (0.553 vs. 0.306. The Kenney extender is effective in preserving the motility, vigor and fertility of stallion semen after 24 hours of cold storage, whereas the Foote extender is not acceptable.

  7. Sustainable transportation according to certification systems: A viability analysis based on neighborhood size and context relevance

    International Nuclear Information System (INIS)

    Gouda, Amr Ah.; Masoumi, Houshmand E.

    2017-01-01

    Urban sustainability certification (USC) systems comprehensively assess and benchmark the sustainability of neighborhoods, communities, etc. However, it is important to understand what USCs mean by neighborhoods and communities in terms of definition and their certified developments' size. This study focuses on sustainable transportation and its measures in USCs in order to discern: how relevant are these measures to the varied local conditions, especially in developing countries? And whether they can generate palpable benefits vis-à-vis the certified developments' site areas. Taking descriptive analysis methods, this study focuses on four prominent USCs, namely, LEED for Neighborhood Development, BREEAM Communities, CASBEE for Urban Development, and The Pearl Community Rating System. The four USCs prescribed multiple measures for sustainable transportation, particularly in favor of active transportation and public transportation. However, the relatively small size of their citified developments and the varying transportation conditions among different contexts attenuate the viability and relevance of the advocated measures for both modes. In order to yield more benefits, USCs should underscore the integrative nature of neighborhoods and communities, and transportation within their prescribed criteria and measures. Moreover, sustainable transportation as a theme should be tailored to the local conditions rather than being adopted or adapted from global USCs. - Highlights: • Transportation measures in four prominent certification systems were investigated. • Most certified neighborhoods, communities and developments have small site areas. • Certifications are insensitive to realities of transport in developing countries. • Benefits from advocating active transport only within certified areas are minimal. • Certifications devalue the integrative nature of neighborhoods and transportation.

  8. Sustainable transportation according to certification systems: A viability analysis based on neighborhood size and context relevance

    Energy Technology Data Exchange (ETDEWEB)

    Gouda, Amr Ah., E-mail: amr.gouda@eng.asu.edu.eg [Center for Technology and Society, Technische Universität Berlin, Hardenbergstr, 16-18, Berlin 10623 (Germany); Architecture Department, Faculty of Engineering, Ain Shams University, 1 Sarayat St., Abassia, Cairo (Egypt); Masoumi, Houshmand E. [Center for Technology and Society, Technische Universität Berlin, Hardenbergstr, 16-18, Berlin 10623 (Germany)

    2017-03-15

    Urban sustainability certification (USC) systems comprehensively assess and benchmark the sustainability of neighborhoods, communities, etc. However, it is important to understand what USCs mean by neighborhoods and communities in terms of definition and their certified developments' size. This study focuses on sustainable transportation and its measures in USCs in order to discern: how relevant are these measures to the varied local conditions, especially in developing countries? And whether they can generate palpable benefits vis-à-vis the certified developments' site areas. Taking descriptive analysis methods, this study focuses on four prominent USCs, namely, LEED for Neighborhood Development, BREEAM Communities, CASBEE for Urban Development, and The Pearl Community Rating System. The four USCs prescribed multiple measures for sustainable transportation, particularly in favor of active transportation and public transportation. However, the relatively small size of their citified developments and the varying transportation conditions among different contexts attenuate the viability and relevance of the advocated measures for both modes. In order to yield more benefits, USCs should underscore the integrative nature of neighborhoods and communities, and transportation within their prescribed criteria and measures. Moreover, sustainable transportation as a theme should be tailored to the local conditions rather than being adopted or adapted from global USCs. - Highlights: • Transportation measures in four prominent certification systems were investigated. • Most certified neighborhoods, communities and developments have small site areas. • Certifications are insensitive to realities of transport in developing countries. • Benefits from advocating active transport only within certified areas are minimal. • Certifications devalue the integrative nature of neighborhoods and transportation.

  9. Collagen-based silver nanoparticles: Study on cell viability, skin permeation, and swelling inhibition

    Energy Technology Data Exchange (ETDEWEB)

    Saura Cardoso, Vinicius, E-mail: vscfisio@ufpi.edu.br [Research Center in Biodiversity and Biotechnology, Biotec, Campus Ministro Reis Velloso, Federal University of Piauí, UFPI, 64202020 Parnaíba, Piauí (Brazil); Physiotherapy Department, Campus Ministro Reis Velloso, Federal University of Piauí, UFPI, 64202020 Parnaíba, Piauí (Brazil); Carvalho Filgueiras, Marcelo de; Medeiros Dutra, Yago; Gomes Teles, Ramon Handerson [Physiotherapy Department, Campus Ministro Reis Velloso, Federal University of Piauí, UFPI, 64202020 Parnaíba, Piauí (Brazil); Morphology and Muscle Physiology Laboratory, LAMFIM, Campus Ministro Reis Velloso, Federal University of Piauí, UFPI, 64202020 Parnaíba, Piauí (Brazil); Rodrigues de Araújo, Alyne [Research Center in Biodiversity and Biotechnology, Biotec, Campus Ministro Reis Velloso, Federal University of Piauí, UFPI, 64202020 Parnaíba, Piauí (Brazil); Primo, Fernando Lucas [Faculdade de Ciências Farmacêuticas, UNESP, Universidade Estadual Paulista, Campus de Araraquara, Departamento de Bioprocessos e Biotecnologia, 14800903 Araraquara, São Paulo (Brazil); Mafud, Ana Carolina; Batista, Larissa Fernandes; Mascarenhas, Yvonne Primerano [Institute of Physics of São Carlos, IFSC, University of São Paulo, USP, 13566590 São Carlos, SP (Brazil); and others

    2017-05-01

    Collagen is considered the most abundant protein in the animal kingdom, comprising 30% of the total amount of proteins and 6% of the human body by weight. Studies that examine the interaction between silver nanoparticles and proteins have been highlighted in the literature in order to understand the stability of the nanoparticle system, the effects observed in biological systems, and the appearance of new chemical pharmaceutical products. The objective of this study was to analyze the behavior of silver nanoparticles stabilized with collagen (AgNPcol) and to check the skin permeation capacity and action in paw edema induced by carrageenan. AgNPcol synthesis was carried out using solutions of reducing agent sodium borohydride (NaBH{sub 4}), silver nitrate (AgNO{sub 3}) and collagen. Characterization was done by using dynamic light scattering (DLS) and X-ray diffraction (XRD) and AFM. Cellular viability testing was performed by using flow cytometry in human melanoma cancer (MV3) and murine fibroblast (L929) cells. The skin permeation study was conducted using a Franz diffusion cell, and the efficiency of AgNPcol against the formation of paw edema in mice was evaluated. The hydrodynamic diameter and zeta potential of AgNPcol were 140.7 ± 7.8 nm and 20.1 ± 0.7 mV, respectively. AgNPcol failed to induce early apoptosis, late apoptosis, and necrosis in L929 cells; however, it exhibited enhanced toxicity in cancer cells (MV3) compared to normal cells (L929). AgNPcol demonstrated increased toxicological effects in cancer MV3 cells, promoting skin permeation, and preventing paw edema. - Highlights: • Silver nanoparticles were synthesized with type I collagen (AgNPcol). • AgNPcol which was characterized by XRD and DLS. • AgNPcol exhibited enhanced toxicity in cancer cells. • The efficiency of the AgNPcol against the paw edema was evaluated.

  10. Collagen-based silver nanoparticles: Study on cell viability, skin permeation, and swelling inhibition

    International Nuclear Information System (INIS)

    Saura Cardoso, Vinicius; Carvalho Filgueiras, Marcelo de; Medeiros Dutra, Yago; Gomes Teles, Ramon Handerson; Rodrigues de Araújo, Alyne; Primo, Fernando Lucas; Mafud, Ana Carolina; Batista, Larissa Fernandes; Mascarenhas, Yvonne Primerano

    2017-01-01

    Collagen is considered the most abundant protein in the animal kingdom, comprising 30% of the total amount of proteins and 6% of the human body by weight. Studies that examine the interaction between silver nanoparticles and proteins have been highlighted in the literature in order to understand the stability of the nanoparticle system, the effects observed in biological systems, and the appearance of new chemical pharmaceutical products. The objective of this study was to analyze the behavior of silver nanoparticles stabilized with collagen (AgNPcol) and to check the skin permeation capacity and action in paw edema induced by carrageenan. AgNPcol synthesis was carried out using solutions of reducing agent sodium borohydride (NaBH 4 ), silver nitrate (AgNO 3 ) and collagen. Characterization was done by using dynamic light scattering (DLS) and X-ray diffraction (XRD) and AFM. Cellular viability testing was performed by using flow cytometry in human melanoma cancer (MV3) and murine fibroblast (L929) cells. The skin permeation study was conducted using a Franz diffusion cell, and the efficiency of AgNPcol against the formation of paw edema in mice was evaluated. The hydrodynamic diameter and zeta potential of AgNPcol were 140.7 ± 7.8 nm and 20.1 ± 0.7 mV, respectively. AgNPcol failed to induce early apoptosis, late apoptosis, and necrosis in L929 cells; however, it exhibited enhanced toxicity in cancer cells (MV3) compared to normal cells (L929). AgNPcol demonstrated increased toxicological effects in cancer MV3 cells, promoting skin permeation, and preventing paw edema. - Highlights: • Silver nanoparticles were synthesized with type I collagen (AgNPcol). • AgNPcol which was characterized by XRD and DLS. • AgNPcol exhibited enhanced toxicity in cancer cells. • The efficiency of the AgNPcol against the paw edema was evaluated.

  11. The economic viability of value-based food chain for dairy farms in mountain regions: an econometric analysis approach

    Directory of Open Access Journals (Sweden)

    Jernej Prišenk

    2016-07-01

    Full Text Available The attention of this paper is drawn to analyze the economic potential of involvement of farmers into the small-medium sized value-based food chain (VBFC. The survey represents a solid dana basis from which econometric modelling approach was further developed. Empirical results reveal the positive economic viability on a general level; this means more stable purchase price of raw milk for dairy farms, which are the part of value-based food chain. Results point at inelastic demand for milk and milk related products. Furthermore, there are some accompanying and underlying indirect social benefits, such as production of high-quality food products, more stable and constant demand for raw milk, steady payments and better social situation. The last one is especially important for the farms operating in less-favored mountain areas where the survey was actually conducted.

  12. Use of luciferase probes to measure ATP in living cells and animals.

    Science.gov (United States)

    Morciano, Giampaolo; Sarti, Alba Clara; Marchi, Saverio; Missiroli, Sonia; Falzoni, Simonetta; Raffaghello, Lizzia; Pistoia, Vito; Giorgi, Carlotta; Di Virgilio, Francesco; Pinton, Paolo

    2017-08-01

    ATP, the energy exchange factor that connects anabolism and catabolism, is required for major reactions and processes that occur in living cells, such as muscle contraction, phosphorylation and active transport. ATP is also the key molecule in extracellular purinergic signaling mechanisms, with an established crucial role in inflammation and several additional disease conditions. Here, we describe detailed protocols to measure the ATP concentration in isolated living cells and animals using luminescence techniques based on targeted luciferase probes. In the presence of magnesium, oxygen and ATP, the protein luciferase catalyzes oxidation of the substrate luciferin, which is associated with light emission. Recombinantly expressed wild-type luciferase is exclusively cytosolic; however, adding specific targeting sequences can modify its cellular localization. Using this strategy, we have constructed luciferase chimeras targeted to the mitochondrial matrix and the outer surface of the plasma membrane. Here, we describe optimized protocols for monitoring ATP concentrations in the cytosol, mitochondrial matrix and pericellular space in living cells via an overall procedure that requires an average of 3 d. In addition, we present a detailed protocol for the in vivo detection of extracellular ATP in mice using luciferase-transfected reporter cells. This latter procedure may require up to 25 d to complete.

  13. Optimal sample size for predicting viability of cabbage and radish seeds based on near infrared spectra of single seeds

    DEFF Research Database (Denmark)

    Shetty, Nisha; Min, Tai-Gi; Gislum, René

    2011-01-01

    The effects of the number of seeds in a training sample set on the ability to predict the viability of cabbage or radish seeds are presented and discussed. The supervised classification method extended canonical variates analysis (ECVA) was used to develop a classification model. Calibration sub......-sets of different sizes were chosen randomly with several iterations and using the spectral-based sample selection algorithms DUPLEX and CADEX. An independent test set was used to validate the developed classification models. The results showed that 200 seeds were optimal in a calibration set for both cabbage...... using all 600 seeds in the calibration set. Thus, the number of seeds in the calibration set can be reduced by up to 67% without significant loss of classification accuracy, which will effectively enhance the cost-effectiveness of NIR spectral analysis. Wavelength regions important...

  14. Extension of landscape-based population viability models to ecoregional scales for conservation planning

    Science.gov (United States)

    Thomas W. Bonnot; Frank R. III Thompson; Joshua Millspaugh

    2011-01-01

    Landscape-based population models are potentially valuable tools in facilitating conservation planning and actions at large scales. However, such models have rarely been applied at ecoregional scales. We extended landscape-based population models to ecoregional scales for three species of concern in the Central Hardwoods Bird Conservation Region and compared model...

  15. Viability of a Web-Based Module for Teaching Electrocardiogram Reading Skills to Psychiatry Residents: Learning Outcomes and Trainee Interest.

    Science.gov (United States)

    DeBonis, Katrina; Blair, Thomas R; Payne, Samuel T; Wigan, Katherine; Kim, Sara

    2015-12-01

    Web-based instruction in post-graduate psychiatry training has shown comparable effectiveness to in-person instruction, but few topics have been addressed in this format. This study sought to evaluate the viability of a web-based curriculum in teaching electrocardiogram (EKG) reading skills to psychiatry residents. Interest in receiving educational materials in this format was also assessed. A web-based curriculum of 41 slides, including eight pre-test and eight post-test questions with emphasis on cardiac complications of psychotropic medications, was made available to all psychiatry residents via email. Out of 57 residents, 30 initiated and 22 completed the module. Mean improvement from pre-test to post-test was 25 %, and all 22 completing participants indicated interest in future web-based instruction. This pilot study suggests that web-based instruction is feasible and under-utilized as a means of teaching psychiatry residents. Potential uses of web-based instruction, such as tracking learning outcomes or patient care longitudinally, are also discussed.

  16. Robustness and management adaptability in tropical rangelands: a viability-based assessment under the non-equilibrium paradigm.

    Science.gov (United States)

    Accatino, F; Sabatier, R; De Michele, C; Ward, D; Wiegand, K; Meyer, K M

    2014-08-01

    Rangelands provide the main forage resource for livestock in many parts of the world, but maintaining long-term productivity and providing sufficient income for the rancher remains a challenge. One key issue is to maintain the rangeland in conditions where the rancher has the greatest possibility to adapt his/her management choices to a highly fluctuating and uncertain environment. In this study, we address management robustness and adaptability, which increase the resilience of a rangeland. After reviewing how the concept of resilience evolved in parallel to modelling views on rangelands, we present a dynamic model of rangelands to which we applied the mathematical framework of viability theory to quantify the management adaptability of the system in a stochastic environment. This quantification is based on an index that combines the robustness of the system to rainfall variability and the ability of the rancher to adjust his/her management through time. We evaluated the adaptability for four possible scenarios combining two rainfall regimes (high or low) with two herding strategies (grazers only or mixed herd). Results show that pure grazing is viable only for high-rainfall regimes, and that the use of mixed-feeder herds increases the adaptability of the management. The management is the most adaptive with mixed herds and in rangelands composed of an intermediate density of trees and grasses. In such situations, grass provides high quantities of biomass and woody plants ensure robustness to droughts. Beyond the implications for management, our results illustrate the relevance of viability theory for addressing the issue of robustness and adaptability in non-equilibrium environments.

  17. Strategic framework for socioeconomic viability of community-based early warning system

    NARCIS (Netherlands)

    Homberg, M.J.C. van den; Posthumus, A.L.

    2014-01-01

    Christian Aid, Cordaid, PVGS and Practical Action established a community-based early warning system for cross-border floods between India and Nepal in 45 Indian villages. The project will scale to 95 villages early 2016. The number of stakeholders and organizational levels of this system with four

  18. Biocorrosion behavior and cell viability of adhesive polymer coated magnesium based alloys for medical implants

    Science.gov (United States)

    Abdal-hay, Abdalla; Dewidar, Montasser; Lim, Jae Kyoo

    2012-11-01

    The present study was ultimately aimed to design novel adhesive biodegradable polymer, poly(vinyl acetate) (PVAc), coatings onto Mg based alloys by the dip-coating technique in order to control the degradation rate and enhance the biocompatibility of magnesium alloys. The influence of various solvents on PVAc surface topography and their protection of Mg alloys were dramatically studied in vitro. Electrochemical polarization, degradation, and PVAc film cytocompatibility were also tested. Our results showed that the solvent had a significant effect on coating quality. PVAc/dichloromethane solution showed a porous structure and solution concentration could control the porous size. The coatings prepared using tetrahydrofuran and dimethylformamide solvents are exceptional in their ability to generate porous morphology even at low polymer concentration. In general, the corrosion performance appears to be different on different PVAc-solvent system. Immersion tests illustrated that the porous morphology on PVAc stabilized corrosion rates. A uniform corrosion attack in artificial simulation body fluid was also exhibited. The cytocompatibility of osteoblast cells (MC3T3) revealed high adherence, proliferation, and survival on the porous structure of PVAc coated Mg alloy, which was not observed for the uncoated samples. This novel PVAc coating is a promising candidate for biodegradable implant materials, which might widen the use of Mg based implants.

  19. Gamma camera based Positron Emission Tomography: a study of the viability on quantification

    International Nuclear Information System (INIS)

    Pozzo, Lorena

    2005-01-01

    Positron Emission Tomography (PET) is a Nuclear Medicine imaging modality for diagnostic purposes. Pharmaceuticals labeled with positron emitters are used and images which represent the in vivo biochemical process within tissues can be obtained. The positron/electron annihilation photons are detected in coincidence and this information is used for object reconstruction. Presently, there are two types of systems available for this imaging modality: the dedicated systems and those based on gamma camera technology. In this work, we utilized PET/SPECT systems, which also allows for the traditional Nuclear Medicine studies based on single photon emitters. There are inherent difficulties which affect quantification of activity and other indices. They are related to the Poisson nature of radioactivity, to radiation interactions with patient body and detector, noise due to statistical nature of these interactions and to all the detection processes, as well as the patient acquisition protocols. Corrections are described in the literature and not all of them are implemented by the manufacturers: scatter, attenuation, random, decay, dead time, spatial resolution, and others related to the properties of each equipment. The goal of this work was to assess these methods adopted by two manufacturers, as well as the influence of some technical characteristics of PET/SPECT systems on the estimation of SUV. Data from a set of phantoms were collected in 3D mode by one camera and 2D, by the other. We concluded that quantification is viable in PET/SPECT systems, including the estimation of SUVs. This is only possible if, apart from the above mentioned corrections, the camera is well tuned and coefficients for sensitivity normalization and partial volume corrections are applied. We also verified that the shapes of the sources used for obtaining these factors play a role on the final results and should be delt with carefully in clinical quantification. Finally, the choice of the region

  20. Biocorrosion behavior and cell viability of adhesive polymer coated magnesium based alloys for medical implants

    Energy Technology Data Exchange (ETDEWEB)

    Abdal-hay, Abdalla [Departmentt of Bionano System Engineering, College of Engineering, Chonbuk National University, Jeonju 561-756 (Korea, Republic of); Department of Mechanical Design Engineering, Advanced wind power system research institute, Chonbuk National University, Jeonju 561-756 (Korea, Republic of); Dewidar, Montasser [Department of Materials and Mechanical Design, Faculty of Energy Engineering, South Valley University, Aswan (Egypt); Lim, Jae Kyoo, E-mail: jklim@jbnu.ac.kr [Department of Mechanical Design Engineering, Advanced wind power system research institute, Chonbuk National University, Jeonju 561-756 (Korea, Republic of)

    2012-11-15

    Highlights: Black-Right-Pointing-Pointer The corrosion behavior of magnesium for orthopedic applications is extremely poor. Black-Right-Pointing-Pointer The solvent (DCM, THF and DMF) had a strong effect on the coatings performance. Black-Right-Pointing-Pointer Mg bar alloy coated with PVAc/DCM layers provided an excellent bonding strength. Black-Right-Pointing-Pointer Treated samples indicated significant damping for the degradation rate. Black-Right-Pointing-Pointer Cytocompatibility on MC3T3 cells of the PVAc/DCM samples revealed a good behavior. - Abstract: The present study was ultimately aimed to design novel adhesive biodegradable polymer, poly(vinyl acetate) (PVAc), coatings onto Mg based alloys by the dip-coating technique in order to control the degradation rate and enhance the biocompatibility of magnesium alloys. The influence of various solvents on PVAc surface topography and their protection of Mg alloys were dramatically studied in vitro. Electrochemical polarization, degradation, and PVAc film cytocompatibility were also tested. Our results showed that the solvent had a significant effect on coating quality. PVAc/dichloromethane solution showed a porous structure and solution concentration could control the porous size. The coatings prepared using tetrahydrofuran and dimethylformamide solvents are exceptional in their ability to generate porous morphology even at low polymer concentration. In general, the corrosion performance appears to be different on different PVAc-solvent system. Immersion tests illustrated that the porous morphology on PVAc stabilized corrosion rates. A uniform corrosion attack in artificial simulation body fluid was also exhibited. The cytocompatibility of osteoblast cells (MC3T3) revealed high adherence, proliferation, and survival on the porous structure of PVAc coated Mg alloy, which was not observed for the uncoated samples. This novel PVAc coating is a promising candidate for biodegradable implant materials, which might

  1. Biocorrosion behavior and cell viability of adhesive polymer coated magnesium based alloys for medical implants

    International Nuclear Information System (INIS)

    Abdal-hay, Abdalla; Dewidar, Montasser; Lim, Jae Kyoo

    2012-01-01

    Highlights: ► The corrosion behavior of magnesium for orthopedic applications is extremely poor. ► The solvent (DCM, THF and DMF) had a strong effect on the coatings performance. ► Mg bar alloy coated with PVAc/DCM layers provided an excellent bonding strength. ► Treated samples indicated significant damping for the degradation rate. ► Cytocompatibility on MC3T3 cells of the PVAc/DCM samples revealed a good behavior. - Abstract: The present study was ultimately aimed to design novel adhesive biodegradable polymer, poly(vinyl acetate) (PVAc), coatings onto Mg based alloys by the dip-coating technique in order to control the degradation rate and enhance the biocompatibility of magnesium alloys. The influence of various solvents on PVAc surface topography and their protection of Mg alloys were dramatically studied in vitro. Electrochemical polarization, degradation, and PVAc film cytocompatibility were also tested. Our results showed that the solvent had a significant effect on coating quality. PVAc/dichloromethane solution showed a porous structure and solution concentration could control the porous size. The coatings prepared using tetrahydrofuran and dimethylformamide solvents are exceptional in their ability to generate porous morphology even at low polymer concentration. In general, the corrosion performance appears to be different on different PVAc–solvent system. Immersion tests illustrated that the porous morphology on PVAc stabilized corrosion rates. A uniform corrosion attack in artificial simulation body fluid was also exhibited. The cytocompatibility of osteoblast cells (MC3T3) revealed high adherence, proliferation, and survival on the porous structure of PVAc coated Mg alloy, which was not observed for the uncoated samples. This novel PVAc coating is a promising candidate for biodegradable implant materials, which might widen the use of Mg based implants.

  2. Two panels of steroid receptor luciferase reporter cell lines for compound profiling

    Czech Academy of Sciences Publication Activity Database

    Sedlák, David; Paguio, A.; Bartůněk, Petr

    2011-01-01

    Roč. 14, č. 2 (2011), s. 248-266 ISSN 1386-2073 R&D Projects: GA MŠk(CZ) LC06077 Institutional research plan: CEZ:AV0Z50520514 Keywords : nuclear hormone receptor * steroid receptor * cell-based luciferase reporter assay Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.785, year: 2011

  3. F-18 Labeled Diabody-Luciferase Fusion Proteins for Optical-ImmunoPET

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Anna M. [Univ. of California, Los Angeles, CA (United States)

    2013-01-18

    The goal of the proposed work is to develop novel dual-labeled molecular imaging probes for multimodality imaging. Based on small, engineered antibodies called diabodies, these probes will be radioactively tagged with Fluorine-18 for PET imaging, and fused to luciferases for optical (bioluminescence) detection. Performance will be evaluated and validated using a prototype integrated optical-PET imaging system, OPET. Multimodality probes for optical-PET imaging will be based on diabodies that are dually labeled with 18F for PET detection and fused to luciferases for optical imaging. 1) Two sets of fusion proteins will be built, targeting the cell surface markers CEA or HER2. Coelenterazine-based luciferases and variant forms will be evaluated in combination with native substrate and analogs, in order to obtain two distinct probes recognizing different targets with different spectral signatures. 2) Diabody-luciferase fusion proteins will be labeled with 18F using amine reactive [18F]-SFB produced using a novel microwave-assisted, one-pot method. 3) Sitespecific, chemoselective radiolabeling methods will be devised, to reduce the chance that radiolabeling will inactivate either the target-binding properties or the bioluminescence properties of the diabody-luciferase fusion proteins. 4) Combined optical and PET imaging of these dual modality probes will be evaluated and validated in vitro and in vivo using a prototype integrated optical-PET imaging system, OPET. Each imaging modality has its strengths and weaknesses. Development and use of dual modality probes allows optical imaging to benefit from the localization and quantitation offered by the PET mode, and enhances the PET imaging by enabling simultaneous detection of more than one probe.

  4. The type and composition of alginate and hyaluronic-based hydrogels influence the viability of stem cells of the apical papilla.

    Science.gov (United States)

    Lambricht, Laure; De Berdt, Pauline; Vanacker, Julie; Leprince, Julian; Diogenes, Anibal; Goldansaz, Hadi; Bouzin, Caroline; Préat, Véronique; Dupont-Gillain, Christine; des Rieux, Anne

    2014-12-01

    The goal of the present work was to evaluate in vitro and in vivo the influence of various types and compositions of natural hydrogels on the viability and metabolic activity of SCAPs. Two alginate, three hyaluronic-based (Corgel™) hydrogel formulations and Matrigel were characterized for their mechanical, surface and microstructure properties using rheology, X-ray photoelectron spectroscopy and scanning electron microscopy, respectively. A characterized SCAP cell line (RP89 cells) was encapsulated in the different experimental hydrogel formulations. Cells were cultured in vitro, or implanted in cyclosporine treated mice. In vitro cell viability was evaluated using a Live/Dead assay and in vitro cellular metabolic activity was evaluated with a MTS assay. In vivo cell apoptosis was evaluated by a TUNEL test and RP89 cells were identified by human mitochondria immunostaining. Hydrogel composition influenced their mechanical and surface properties, and their microstructure. In vitro cell viability was above 80% after 2 days but decreased significantly after 7 days (60-40%). Viability at day 7 was the highest in Matrigel (70%) and then in Corgel 1.5 (60%). Metabolic activity increased over time in all the hydrogels, excepted in alginate SLM. SCAPs survived after 1 week in vivo with low apoptosis (<1%). The highest number of RP89 cells was found in Corgel 5.5 (140cells/mm(2)). Collectively, these data demonstrate that SCAP viability was directly modulated by hydrogel composition and suggest that a commercially available hyaluronic acid-based formulation might be a suitable delivery vehicle for SCAP-based dental pulp regeneration strategies. Copyright © 2014 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

  5. Luciferase inactivation in the luminous marine bacterium Vibrio harveyi.

    Science.gov (United States)

    Reeve, C A; Baldwin, T O

    1981-06-01

    Luciferase was rapidly inactivated in stationary-phase cultures of the wild type of the luminous marine bacterium Vibrio harveyi, but was stable in stationary-phase cultures of mutants of V. harveyi that are nonluminous without exogenous aldehyde, termed the aldehyde-deficient mutants. The inactivation in the wild type was halted by cell lysis and was slowed or stopped by O2 deprivation or by addition of KCN and NaF or of chloramphenicol. If KCN and NaF or chloramphenicol were added to a culture before the onset of luciferase inactivation, then luciferase inactivation did not occur. However, if these inhibitors were added after the onset of luciferase inactivation, then luciferase inactivation continued for about 2 to 3 h before the inactivation process stopped. The onset of luciferase inactivation in early stationary-phase cultures of wild-type cell coincided with a slight drop in the intracellular adenosine 5'-triphosphate (ATP) level from a relatively constant log-phase value of 20 pmol of ATP per microgram of soluble cell protein. Addition of KCN and NaF to a culture shortly after this drop in ATP caused a rapid decrease in the ATP level to about 4 pmol of ATP per microgram whereas chloramphenicol added at this same time caused a transient increase in ATP level to about 25 pmol/microgram. The aldehyde-deficient mutant (M17) showed a relatively constant log-phase ATP level identical with that of the wild-type cells, but rather than decreasing in early stationary phase, the ATP level increased to a value twice that in log-phase cells. We suggest that the inactivation of luciferase is dependent on the synthesis of some factor which is produced during stationary phase and is itself unstable, and whose synthesis is blocked by chloramphenicol or cyanide plus fluoride.

  6. Flexible Measurement of Bioluminescent Reporters Using an Automated Longitudinal Luciferase Imaging Gas- and Temperature-optimized Recorder (ALLIGATOR).

    Science.gov (United States)

    Crosby, Priya; Hoyle, Nathaniel P; O'Neill, John S

    2017-12-13

    Luciferase-based reporters of cellular gene expression are in widespread use for both longitudinal and end-point assays of biological activity. In circadian rhythms research, for example, clock gene fusions with firefly luciferase give rise to robust rhythms in cellular bioluminescence that persist over many days. Technical limitations associated with photomultiplier tubes (PMT) or conventional microscopy-based methods for bioluminescence quantification have typically demanded that cells and tissues be maintained under quite non-physiological conditions during recording, with a trade-off between sensitivity and throughput. Here, we report a refinement of prior methods that allows long-term bioluminescence imaging with high sensitivity and throughput which supports a broad range of culture conditions, including variable gas and humidity control, and that accepts many different tissue culture plates and dishes. This automated longitudinal luciferase imaging gas- and temperature-optimized recorder (ALLIGATOR) also allows the observation of spatial variations in luciferase expression across a cell monolayer or tissue, which cannot readily be observed by traditional methods. We highlight how the ALLIGATOR provides vastly increased flexibility for the detection of luciferase activity when compared with existing methods.

  7. The Impact of a Values-Based Supply Chain (VBSC on Farm-Level Viability, Sustainability and Resilience: Case Study Evidence

    Directory of Open Access Journals (Sweden)

    Teresa Hooks

    2017-02-01

    Full Text Available The ‘Agriculture of the Middle’ (AotM development paradigm emphasises that in order to survive, family farms must transition from a supply chain approach to a values-based supply chain (VBSC approach, involving amendments to both product type and actor dynamics within the chain. This paper presents a qualitative case study of a beef co-operative integrated to a VBSC. We use an analytical framework of viability, sustainability and resilience to analyse impacts at farm-level. Our analysis highlights a number of positive effects on farm-level viability, sustainability and resilience. These benefits stemmed largely from improvements to market orientation, price stability, and members’ capacities in responding to problems. However, the autonomy of the co-operative was challenged by VBSC chain members, which impacted negatively on the stability of the co-operative.

  8. A new image-based tool for the high throughput phenotyping of pollen viability: evaluation of inter- and intra-cultivar diversity in grapevine.

    Science.gov (United States)

    Tello, Javier; Montemayor, María Ignacia; Forneck, Astrid; Ibáñez, Javier

    2018-01-01

    Low pollen viability may limit grapevine yield under certain conditions, causing relevant economic losses to grape-growers. It is usually evaluated by the quantification of the number of viable and non-viable pollen grains that are present in a sample after an adequate pollen grain staining procedure. Although the manual counting of both types of grains is the simplest and most sensitive approach, it is a laborious and time-demanding process. In this regard, novel image-based approaches can assist in the objective, accurate and cost-effective phenotyping of this trait. Here, we introduce PollenCounter, an open-source macro implemented as a customizable Fiji tool for the high-throughput phenotyping of pollen viability. This tool splits RGB images of stained pollen grains into its primary channels, retaining red and green color fractionated images (which contain information on total and only viable pollen grains, respectively) for the subsequent isolation and counting of the regions of interest (pollen grains). This framework was successfully used for the analysis of pollen viability of a high number of samples collected in a large collection of grapevine cultivars. Results revealed a great genetic variability, from cultivars having very low pollen viability (like Corinto Bianco; viability: 14.1 ± 1.3%) to others with a very low presence of sterile pollen grains (Cuelga; viability: 98.2 ± 0.5%). A wide range of variability was also observed among several clones of cv. Tempranillo Tinto (from 97.9 ± 0.9 to 60.6 ± 5.9%, in the first season). Interestingly, the evaluation of this trait in a second season revealed differential genotype-specific sensitivity to environment. The use of PollenCounter is expected to aid in different areas, including genetics research studies, crop improvement and breeding strategies that need of fast, precise and accurate results. Considering its flexibility, it can be used not only in grapevine, but also in other species showing

  9. Development and application of hepatitis C reporter viruses with genotype 1 to 7 core-nonstructural protein 2 (NS2) expressing fluorescent proteins or luciferase in modified JFH1 NS5A

    DEFF Research Database (Denmark)

    Gottwein, Judith M; Jensen, Tanja B; Mathiesen, Christian K

    2011-01-01

    to 2a(J6) tagged with EGFP, DsRed-Express2, mCherry, or Renilla luciferase (RLuc), yielding peak supernatant infectivity titers of 4 to 5 log(10) focus-forming units (FFU)/ml. 2a(J6) with ¿40 or ¿25 was fully viable in Huh7.5 cells. In human liver chimeric mice, 2a(J6)-EGFP¿40 acquired various...... deletions in EGFP, while 2a(J6)¿40 did not show an impaired viability. We further developed panels of JFH1-based genotype 1 to 7 core-NS2 recombinants expressing EGFP- or RLuc-NS5A¿40 fusion proteins. In cell culture, the different EGFP recombinants showed growth characteristics comparable to those...

  10. Highly specific expression of luciferase gene in lungs of naive nude mice directed by prostate-specific antigen promoter

    International Nuclear Information System (INIS)

    Li Hongwei; Li Jinzhong; Helm, Gregory A.; Pan Dongfeng

    2005-01-01

    PSA promoter has been demonstrated the utility for tissue-specific toxic gene therapy in prostate cancer models. Characterization of foreign gene overexpression in normal animals elicited by PSA promoter should help evaluate therapy safety. Here we constructed an adenovirus vector (AdPSA-Luc), containing firefly luciferase gene under the control of the 5837 bp long prostate-specific antigen promoter. A charge coupled device video camera was used to non-invasively image expression of firefly luciferase in nude mice on days 3, 7, 11 after injection of 2 x 10 9 PFU of AdPSA-Luc virus via tail vein. The result showed highly specific expression of the luciferase gene in lungs of mice from day 7. The finding indicates the potential limitations of the suicide gene therapy of prostate cancer based on selectivity of PSA promoter. By contrary, it has encouraging implications for further development of vectors via PSA promoter to enable gene therapy for pulmonary diseases

  11. Possible Overestimation of Surface Disinfection Efficiency by Assessment Methods Based on Liquid Sampling Procedures as Demonstrated by In Situ Quantification of Spore Viability

    Science.gov (United States)

    Grand, I.; Bellon-Fontaine, M.-N.; Herry, J.-M.; Hilaire, D.; Moriconi, F.-X.; Naïtali, M.

    2011-01-01

    The standard test methods used to assess the efficiency of a disinfectant applied to surfaces are often based on counting the microbial survivors sampled in a liquid, but total cell removal from surfaces is seldom achieved. One might therefore wonder whether evaluations of microbial survivors in liquid-sampled cells are representative of the levels of survivors in whole populations. The present study was thus designed to determine the “damaged/undamaged” status induced by a peracetic acid disinfection for Bacillus atrophaeus spores deposited on glass coupons directly on this substrate and to compare it to the status of spores collected in liquid by a sampling procedure. The method utilized to assess the viability of both surface-associated and liquid-sampled spores included fluorescence labeling with a combination of Syto 61 and Chemchrome V6 dyes and quantifications by analyzing the images acquired by confocal laser scanning microscopy. The principal result of the study was that the viability of spores sampled in the liquid was found to be poorer than that of surface-associated spores. For example, after 2 min of peracetic acid disinfection, less than 17% ± 5% of viable cells were detected among liquid-sampled cells compared to 79% ± 5% or 47% ± 4%, respectively, when the viability was evaluated on the surface after or without the sampling procedure. Moreover, assessments of the survivors collected in the liquid phase, evaluated using the microscopic method and standard plate counts, were well correlated. Evaluations based on the determination of survivors among the liquid-sampled cells can thus overestimate the efficiency of surface disinfection procedures. PMID:21742922

  12. Possible overestimation of surface disinfection efficiency by assessment methods based on liquid sampling procedures as demonstrated by in situ quantification of spore viability.

    Science.gov (United States)

    Grand, I; Bellon-Fontaine, M-N; Herry, J-M; Hilaire, D; Moriconi, F-X; Naïtali, M

    2011-09-01

    The standard test methods used to assess the efficiency of a disinfectant applied to surfaces are often based on counting the microbial survivors sampled in a liquid, but total cell removal from surfaces is seldom achieved. One might therefore wonder whether evaluations of microbial survivors in liquid-sampled cells are representative of the levels of survivors in whole populations. The present study was thus designed to determine the "damaged/undamaged" status induced by a peracetic acid disinfection for Bacillus atrophaeus spores deposited on glass coupons directly on this substrate and to compare it to the status of spores collected in liquid by a sampling procedure. The method utilized to assess the viability of both surface-associated and liquid-sampled spores included fluorescence labeling with a combination of Syto 61 and Chemchrome V6 dyes and quantifications by analyzing the images acquired by confocal laser scanning microscopy. The principal result of the study was that the viability of spores sampled in the liquid was found to be poorer than that of surface-associated spores. For example, after 2 min of peracetic acid disinfection, less than 17% ± 5% of viable cells were detected among liquid-sampled cells compared to 79% ± 5% or 47% ± 4%, respectively, when the viability was evaluated on the surface after or without the sampling procedure. Moreover, assessments of the survivors collected in the liquid phase, evaluated using the microscopic method and standard plate counts, were well correlated. Evaluations based on the determination of survivors among the liquid-sampled cells can thus overestimate the efficiency of surface disinfection procedures.

  13. Excited-State Dynamics of Oxyluciferin in Firefly Luciferase

    KAUST Repository

    Snellenburg, Joris J.

    2016-11-23

    The color variations of light emitted by some natural and mutant luciferases are normally attributed to collective factors referred to as microenvironment effects; however, the exact nature of these interactions between the emitting molecule (oxyluciferin) and the active site remains elusive. Although model studies of noncomplexed oxyluciferin and its variants have greatly advanced the understanding of its photochemistry, extrapolation of the conclusions to the real system requires assumptions about the polarity and proticity of the active site. To decipher the intricate excited-state dynamics, global and target analysis is performed here for the first time on the steady-state and time-resolved spectra of firefly oxyluciferin complexed with luciferase from the Japanese firefly (Luciola cruciata). The experimental steady-state and time resolved luminescence spectra of the oxyluciferin/luciferase complex in solution are compared with the broadband time-resolved firefly bioluminescence recorded in vivo. The results demonstrate that de-excitation of the luminophore results in a complex cascade of photoinduced proton transfer processes and can be interpreted by the pH dependence of the emitted light. It is confirmed that proton transfer is the central event in the spectrochemistry of this system for which any assignment of the pH dependent emission to a single chemical species would be an oversimplification.

  14. Excited-State Dynamics of Oxyluciferin in Firefly Luciferase

    KAUST Repository

    Snellenburg, Joris J.; Laptenok, Sergey P.; DeSa, Richard J.; Naumov, Pance; Solntsev, Kyril M.

    2016-01-01

    The color variations of light emitted by some natural and mutant luciferases are normally attributed to collective factors referred to as microenvironment effects; however, the exact nature of these interactions between the emitting molecule (oxyluciferin) and the active site remains elusive. Although model studies of noncomplexed oxyluciferin and its variants have greatly advanced the understanding of its photochemistry, extrapolation of the conclusions to the real system requires assumptions about the polarity and proticity of the active site. To decipher the intricate excited-state dynamics, global and target analysis is performed here for the first time on the steady-state and time-resolved spectra of firefly oxyluciferin complexed with luciferase from the Japanese firefly (Luciola cruciata). The experimental steady-state and time resolved luminescence spectra of the oxyluciferin/luciferase complex in solution are compared with the broadband time-resolved firefly bioluminescence recorded in vivo. The results demonstrate that de-excitation of the luminophore results in a complex cascade of photoinduced proton transfer processes and can be interpreted by the pH dependence of the emitted light. It is confirmed that proton transfer is the central event in the spectrochemistry of this system for which any assignment of the pH dependent emission to a single chemical species would be an oversimplification.

  15. Distinction of Gram-positive and -negative bacteria using a colorimetric microbial viability assay based on the reduction of water-soluble tetrazolium salts with a selection medium.

    Science.gov (United States)

    Tsukatani, Tadayuki; Suenaga, Hikaru; Higuchi, Tomoko; Shiga, Masanobu; Noguchi, Katsuya; Matsumoto, Kiyoshi

    2011-01-01

    Bacteria are fundamentally divided into two groups: Gram-positive and Gram-negative. Although the Gram stain and other techniques can be used to differentiate these groups, some issues exist with traditional approaches. In this study, we developed a method for differentiating Gram-positive and -negative bacteria using a colorimetric microbial viability assay based on the reduction of the tetrazolium salt {2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt} (WST-8) via 2-methyl-1,4-napthoquinone with a selection medium. We optimized the composition of the selection medium to allow the growth of Gram-negative bacteria while inhibiting the growth of Gram-positive bacteria. When the colorimetric viability assay was carried out in a selection medium containing 0.5µg/ml crystal violet, 5.0 µg/ml daptomycin, and 5.0µg/ml vancomycin, the reduction in WST-8 by Gram-positive bacteria was inhibited. On the other hand, Gram-negative bacteria produced WST-8-formazan in the selection medium. The proposed method was also applied to determine the Gram staining characteristics of bacteria isolated from various foodstuffs. There was good agreement between the results obtained using the present method and those obtained using a conventional staining method. These results suggest that the WST-8 colorimetric assay with selection medium is a useful technique for accurately differentiating Gram-positive and -negative bacteria.

  16. In Vivo Imaging of Retinoic Acid Receptor Activity using a Sodium/Iodide Symporter and Luciferase Dual Imaging Reporter Gene

    Directory of Open Access Journals (Sweden)

    Min Kyung So

    2004-07-01

    Full Text Available Retinoic acids are natural derivatives of vitamin A, and play important roles in modulating tumor cell growth by regulating differentiation, thus suggesting the potential use of these derivatives in cancer therapy and prevention. To visualize the intranuclear responses of functional retinoic acid receptors, we have developed a dual-imaging reporter gene system based on the use of sodium/iodide symporter (NIS and luciferase in cancer cell lines. NIS and luciferase genes were linked with an internal ribosome entry site, and placed under the control of an artificial cis-acting retinoic acid responsive element (pRARE/NL. After retinoic acid treatment, I-125 uptake by pRARE/NL transfected cells was found to have increased by up to about five times that of nontreated cells. The bioluminescence intensity of pRARE/NL transfected cells showed dose-dependency. In vivo luciferase images showed higher intensity in retinoic acid treated SK-RARE/NL tumors, and scintigraphic images of SK-RARE/NL tumors showed increased Tc-99m uptake after retinoic acid treatment. The NIS/luciferase imaging reporter system was sufficiently sensitive to allow the visualization of intranuclear retinoic acid receptor activity. This cis-enhancer imaging reporter system may be useful in vitro and in vivo for the evaluation of retinoic acid responses in such areas as cellular differentiation and chemoprevention.

  17. Cellular Immune Response Against Firefly Luciferase After Sleeping Beauty–Mediated Gene Transfer In Vivo

    Science.gov (United States)

    Podetz-Pedersen, Kelly M.; Vezys, Vaiva; Somia, Nikunj V.; Russell, Stephen J.

    2014-01-01

    Abstract The Sleeping Beauty (SB) transposon system has been shown to mediate new gene sequence integration resulting in long-term expression. Here the effectiveness of hyperactive SB100X transposase was tested, and we found that hydrodynamic co-delivery of a firefly luciferase transposon (pT2/CaL) along with SB100X transposase (pCMV-SB100X) resulted in remarkably sustained, high levels of luciferase expression. However, after 4 weeks there was a rapid, animal-by-animal loss of luciferase expression that was not observed in immunodeficient mice. We hypothesized that this sustained, high-level luciferase expression achieved using the SB100X transposase elicits an immune response in pT2/CaL co-administered mice, which was supported by the rapid loss of luciferase expression upon challenge of previously treated animals and in naive animals adoptively transferred with splenocytes from previously treated animals. Specificity of the immune response to luciferase was demonstrated by increased cytokine expression in splenocytes after exposure to luciferase peptide in parallel with MHC I–luciferase peptide tetramer binding. This anti-luciferase immune response observed following continuous, high-level luciferase expression in vivo clearly impacts its use as an in vivo reporter. As both an immunogen and an extremely sensitive reporter, luciferase is also a useful model system for the study of immune responses following in vivo gene transfer and expression. PMID:25093708

  18. Increased level of extracellular ATP at tumor sites: in vivo imaging with plasma membrane luciferase.

    Directory of Open Access Journals (Sweden)

    Patrizia Pellegatti

    2008-07-01

    Full Text Available There is growing awareness that tumour cells build up a "self-advantageous" microenvironment that reduces effectiveness of anti-tumour immune response. While many different immunosuppressive mechanisms are likely to come into play, recent evidence suggests that extracellular adenosine acting at A2A receptors may have a major role in down-modulating the immune response as cancerous tissues contain elevated levels of adenosine and adenosine break-down products. While there is no doubt that all cells possess plasma membrane adenosine transporters that mediate adenosine uptake and may also allow its release, it is now clear that most of extracellularly-generated adenosine originates from the catabolism of extracellular ATP.Measurement of extracellular ATP is generally performed in cell supernatants by HPLC or soluble luciferin-luciferase assay, thus it generally turns out to be laborious and inaccurate. We have engineered a chimeric plasma membrane-targeted luciferase that allows in vivo real-time imaging of extracellular ATP. With this novel probe we have measured the ATP concentration within the tumour microenvironment of several experimentally-induced tumours.Our results show that ATP in the tumour interstitium is in the hundreds micromolar range, while it is basically undetectable in healthy tissues. Here we show that a chimeric plasma membrane-targeted luciferase allows in vivo detection of high extracellular ATP concentration at tumour sites. On the contrary, tumour-free tissues show undetectable extracellular ATP levels. Extracellular ATP may be crucial for the tumour not only as a stimulus for growth but also as a source of an immunosuppressive agent such as adenosine. Our approach offers a new tool for the investigation of the biochemical composition of tumour milieu and for development of novel therapies based on the modulation of extracellular purine-based signalling.

  19. Multifractal resilience and viability

    Science.gov (United States)

    Tchiguirinskaia, I.; Schertzer, D. J. M.

    2017-12-01

    The term resilience has become extremely fashionable and there had been many attempts to provide operational definition and in fact metrics going beyond a set of more or less ad-hoc indicators. The viability theory (Aubin and Saint-Pierre, 2011) have been used to give a rather precise mathematical definition of resilience (Deffuant and Gilbert, 2011). However, it does not grasp the multiscale nature of resilience that is rather fundamental as particularly stressed by Folke et al (2010). In this communication, we first recall a preliminary attempt (Tchiguirinskaia et al., 2014) to define multifractal resilience with the help of the maximal probable singularity. Then we extend this multifractal approach to the capture basin of the viability, therefore the resilient basin. Aubin, J P, A. Bayen, and P Saint-Pierre (2011). Viability Theory. New Directions. Springer, Berlin,. Deffuant, G. and Gilbert, N. (eds) (2011) Viability and Resilience of Complex Systems. Springer Berlin.Folke, C., S R Carpenter, B Walker, M Sheffer, T Chapin, and J Rockstroem (2010). Resilience thinking: integrating re- silience, adaptability and transformability. Ecology and So- ciety, 14(4):20, Tchiguirinskaia,I., D. Schertzer, , A. Giangola-Murzyn and T. C. Hoang (2014). Multiscale resilience metrics to assess flood. Proceedings of ICCSA 2014, Normandie University, Le Havre, France -.

  20. Viability, invariance and applications

    CERN Document Server

    Carja, Ovidiu; Vrabie, Ioan I

    2007-01-01

    The book is an almost self-contained presentation of the most important concepts and results in viability and invariance. The viability of a set K with respect to a given function (or multi-function) F, defined on it, describes the property that, for each initial data in K, the differential equation (or inclusion) driven by that function or multi-function) to have at least one solution. The invariance of a set K with respect to a function (or multi-function) F, defined on a larger set D, is that property which says that each solution of the differential equation (or inclusion) driven by F and issuing in K remains in K, at least for a short time.The book includes the most important necessary and sufficient conditions for viability starting with Nagumo's Viability Theorem for ordinary differential equations with continuous right-hand sides and continuing with the corresponding extensions either to differential inclusions or to semilinear or even fully nonlinear evolution equations, systems and inclusions. In th...

  1. Fluoro-luminometric real-time measurement of bacterial viability and killing.

    Science.gov (United States)

    Lehtinen, Janne; Virta, Marko; Lilius, Esa Matti

    2003-10-01

    The viability and killing of Escherichia coli was measured on a real-time basis using a fluoro-luminometric device, which allows successive measurements of fluorescence and bioluminescence without user intervention. Bacteria were made fluorescent and bioluminescent by expression of gfp and insect luciferase (lucFF) genes. The green fluorescent protein (GFP) is a highly fluorescent, extremely stable protein, which accumulates in cells during growth, and therefore the measured fluorescence signal was proportional to the total number of cells. The luciferase reaction is dependent of ATP produced by living cells, so that the bioluminescence level was a direct measure of the viable cells. In contrast to the bacterial luciferase, the insect luciferase uses a water-soluble and nonvolatile substrate, which makes automated multi-well microplate assay possible. For the validation of the assay, the proportion of living and dead cell populations was experimentally modified by incubating E. coli cells in the presence of various ethanol concentrations. Bacterial viability and killing measured by a fluoro-luminometric assay correlated fairly well with the reference methods: conventional plate counting, optical density measurement and various flow cytometric analyses. The real-time assay described here allows following the changes in bacterial cultures and assessing the bactericidal and other effects of various chemical, immunological and physical agents simultaneously in large numbers of samples.

  2. Application of luciferase assay for ATP to antimicrobial drug susceptibility

    Science.gov (United States)

    Chappelle, E. W.; Picciolo, G. L.; Vellend, H.; Tuttle, S. A.; Barza, M. J.; Weinstein, L. (Inventor)

    1977-01-01

    The susceptibility of bacteria, particularly those derived from body fluids, to antimicrobial agents is determined in terms of an ATP index measured by culturing a bacterium in a growth medium. The amount of ATP is assayed in a sample of the cultured bacterium by measuring the amount of luminescent light emitted when the bacterial ATP is reacted with a luciferase-luciferin mixture. The sample of the cultured bacterium is subjected to an antibiotic agent. The amount of bacterial adenosine triphosphate is assayed after treatment with the antibiotic by measuring the luminescent light resulting from the reaction. The ATP index is determined from the values obtained from the assay procedures.

  3. Terminology for pregnancy loss prior to viability

    DEFF Research Database (Denmark)

    Kolte, A M; Bernardi, L A; Christiansen, O B

    2015-01-01

    Pregnancy loss prior to viability is common and research in the field is extensive. Unfortunately, terminology in the literature is inconsistent. The lack of consensus regarding nomenclature and classification of pregnancy loss prior to viability makes it difficult to compare study results from...... different centres. In our opinion, terminology and definitions should be based on clinical findings, and when possible, transvaginal ultrasound. With this Early Pregnancy Consensus Statement, it is our goal to provide clear and consistent terminology for pregnancy loss prior to viability....

  4. Agent-based model for electricity consumption and storage to evaluate economic viability of tariff arbitrage for residential sector demand response

    International Nuclear Information System (INIS)

    Zheng, Menglian; Meinrenken, Christoph J.; Lackner, Klaus S.

    2014-01-01

    Highlights: • Storage-based demand response (loadshifting) is underutilized in residential sector. • Economics (arbitrage savings versus equipment cost) are not well understood. • Stochastic demand models and real-life tariffs can illuminate economic viability. • A range of available storage options provide economically viable DR. • Daily/seasonal stochastic demand variations crucial to understanding optimum capacity. - Abstract: Demand response (DR) is one of many approaches to address temporal mismatches in demand and supply of grid electricity. More common in the commercial sector, DR usually refers to reducing consumption at certain hours or seasons, thus reducing peak demand from the grid. In the residential sector, where sophisticated appliance-level controls such as automatic dimming of lights or on-demand lowering of air conditioning are less common, building-based electricity storage to shift grid consumption from peak to off-peak times could provide DR without requiring consumers to operate their appliances on shifted or reduced schedules: Storage would be dispatched to appliances as needed while still shaving peaks on the grid. Technologically, storage and two-way-inverters are readily available to enable such residential DR. Economically, however, the situation is less clear. Specifically, are time-varying electricity tariffs available such that electricity cost reduction via arbitrage could offset manufacturing, financing, and installation costs of the required storage? To address this question we (i) devise an agent-based appliance-level stochastic model to simulate the electricity demand of an average U.S. household; (ii) loadshift the demand via simple dispatch strategies; and (iii) determine potential profits to the building owner, i.e. reduced electricity cost of the modified demand with realistic tariffs (Con Edison, NY) minus storage cost. We determine the economic viability for a range of traditional and advanced storage technologies

  5. In vivo bioimaging with tissue-specific transcription factor activated luciferase reporters.

    OpenAIRE

    Buckley, SM; Delhove, JM; Perocheau, DP; Karda, R; Rahim, AA; Howe, SJ; Ward, NJ; Birrell, MA; Belvisi, MG; Arbuthnot, P; Johnson, MR; Waddington, SN; McKay, TR

    2015-01-01

    The application of transcription factor activated luciferase reporter cassettes in vitro is widespread but potential for in vivo application has not yet been realized. Bioluminescence imaging enables non-invasive tracking of gene expression in transfected tissues of living rodents. However the mature immune response limits luciferase expression when delivered in adulthood. We present a novel approach of tissue-targeted delivery of transcription factor activated luciferase reporter lentiviruse...

  6. Luciferase-Specific Coelenterazine Analogues for Optical Contamination-Free Bioassays

    OpenAIRE

    Ryo Nishihara; Masahiro Abe; Shigeru Nishiyama; Daniel Citterio; Koji Suzuki; Sung Bae Kim

    2017-01-01

    Spectral overlaps among the multiple optical readouts commonly cause optical contamination in fluorescence and bioluminescence. To tackle this issue, we created five-different lineages of coelenterazine (CTZ) analogues designed to selectively illuminate a specific luciferase with unique luciferase selectivity. In the attempt, we found that CTZ analogues with ethynyl or styryl groups display dramatically biased bioluminescence to specific luciferases and pHs by modifying the functional groups ...

  7. [Morbidity and mortality in newborns at the limit of viability in Spain: a population-based study].

    Science.gov (United States)

    García-Muñoz Rodrigo, F; García-Alix Pérez, A; García Hernández, J A; Figueras Aloy, J

    2014-06-01

    Perinatal care in extremely immature newborns is a clinical and ethical problem of great importance for professionals and families, and requires that the available information on the chances of child survival is of the highest quality. The aim of this study was to determine the specific rates of survival at hospital discharge, and survival without major morbidity in newborns with a gestation age (GA) ≤ 26 weeks in Spain. We included live newborns ≤ 26 weeks admitted to the collaborating centers of the SEN1500 network (2004-2010). Out born patients, infants who died in delivery room, and those with congenital anomalies incompatible with life were excluded. A total of 3,236 patients were included. GA specific survival was 12.5, 13.1, 36.9, 55.7, and 71.9% at 22, 23, 24, 25, and 26 weeks of GA, respectively. Survival without severe intracranial hemorrhage, periventricular leukomalacia, bronchopulmonary dysplasia, and/or retinopathy of prematurity was 1.5, 9.5, 19.0, and 29.9% at 23, 24, 25 and 26 weeks GA, respectively. Survival without major morbidity in infants less than 23 weeks GA is exceptional, and scarce in newborns with 23 and 24 weeks GA. Infants ≥ 25 weeks GA have reasonable chances of survival and, in the absence of major malformations or other relevant complications, should be offered active resuscitation and intensive care. The continuous updating of the results of individual centers is of utmost importance, as well as their comparison with the reference population-based results. Copyright © 2013 Asociación Española de Pediatría. Published by Elsevier Espana. All rights reserved.

  8. Determination of water-soluble vitamins using a colorimetric microbial viability assay based on the reduction of water-soluble tetrazolium salts.

    Science.gov (United States)

    Tsukatani, Tadayuki; Suenaga, Hikaru; Ishiyama, Munetaka; Ezoe, Takatoshi; Matsumoto, Kiyoshi

    2011-07-15

    A method for the determination of water-soluble vitamins using a colorimetric microbial viability assay based on the reduction of the tetrazolium salt {2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8)} via 2-methyl-1,4-napthoquinone (NQ) was developed. Measurement conditions were optimized for the microbiological determination of water-soluble vitamins, such as vitamin B(6), biotin, folic acid, niacin, and pantothenic acid, using microorganisms that have a water-soluble vitamin requirement. A linear relationship between absorbance and water-soluble vitamin concentration was obtained. The proposed method was applied to determine the concentration of vitamin B(6) in various foodstuffs. There was good agreement between vitamin B(6) concentrations determined after 24h using the WST-8 colorimetric method and those obtained after 48h using a conventional method. The results suggest that the WST-8 colorimetric assay is a useful method for the rapid determination of water-soluble vitamins in a 96-well microtiter plate. Copyright © 2011 Elsevier Ltd. All rights reserved.

  9. Viability Assessment Volume 1

    International Nuclear Information System (INIS)

    1998-01-01

    Since May 1996, under its draft Civilian Radioactive Waste Management Program Plan (DOE 1996), DOE has been carrying out a 5-year program of work to support the decision in 2001 by the Secretary of Energy on whether or not to recommend the site to the President. Part of this program was to address major unresolved technical issues and to complete an assessment of the viability of the Yucca Mountain site by 1998. Affirming the DOE plans, Congress directed DOE in the 1997 Energy and Water Development Appropriations Act to provide a viability assessment of the Yucca Mountain site to Congress and the President. This Viability Assessment (VA) document is the DOE report to Congress and the President. They are expected to use the VA to make an informed decision about program direction and funding. Drawing on 15 years of scientific investigation and design work at Yucca Mountain, the VA summarizes a large technical basis of field investigations, laboratory tests, models, analyses, and engineering, described in cited references. The VA identifies the major uncertainties relevant to the technical defensibility of DOE analyses and designs, the DOE approach to managing these uncertainties, and the status of work toward the site recommendation and LA. The VA also identifies DOE plans for the remaining work, and the estimated costs of completing an LA and constructing and operating a repository. The attention to uncertainties is important because DOE must evaluate how the repository will perform during the next 10,000 years or longer. Uncertainties exist because of variability in the natural (geologic and hydrologic) systems at Yucca Mountain and because of imperfect scientific understanding of the natural processes that might affect the repository system. This is Volume 1 and it covers, Introduction and Site Characteristics, includes a high-level summary of the results of the VA and some additional background information. (The overview is bound separately.) Section 1 of Volume

  10. A novel firefly luciferase biosensor enhances the detection of apoptosis induced by ESAT-6 family proteins of Mycobacterium tuberculosis

    International Nuclear Information System (INIS)

    Shi, Junwei; Zhang, Huan; Fang, Liurong; Xi, Yongqiang; Zhou, Yanrong; Luo, Rui; Wang, Dang; Xiao, Shaobo; Chen, Huanchun

    2014-01-01

    Highlights: • We developed a novel firefly luciferase based biosensor to detect apoptosis. • The novel biosensor 233-DnaE-DEVDG was reliable, sensitive and convenient. • 233-DnaE-DEVDG faithfully indicated ESAT-6 family proteins of Mycobacterium tuberculosis induced apoptosis. • EsxA, esxT and esxL in ESAT-6 family proteins induced apoptosis. • Activation of nuclear factor-κB (NF-κB) participated in esxT-induced apoptosis. - Abstract: The activation of caspase-3 is a key surrogate marker for detecting apoptosis. To quantitate caspase-3 activity, we constructed a biosensor comprising a recombinant firefly luciferase containing a caspase-3 cleavage site. When apoptosis was induced, caspase-3 cleavage of the biosensor activated firefly luciferase by a factor greater than 25. The assay conveniently detected apoptosis in real time, indicating that it will facilitate drug discovery. We screened ESAT-6 family proteins of Mycobacterium tuberculosis and found that esxA, esxT and esxL induced apoptosis. Further, activation of nuclear factor-κB (NF-κB) and the NF-κB-regulated genes encoding tumor necrosis factor-α (TNF-α) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) participated in esxT-induced apoptosis. We conclude that this assay is useful for high-throughput screening to identify and characterize proteins and drugs that regulate apoptosis

  11. A novel firefly luciferase biosensor enhances the detection of apoptosis induced by ESAT-6 family proteins of Mycobacterium tuberculosis

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Junwei; Zhang, Huan; Fang, Liurong; Xi, Yongqiang; Zhou, Yanrong; Luo, Rui; Wang, Dang, E-mail: wangdang511@126.com; Xiao, Shaobo; Chen, Huanchun

    2014-10-03

    Highlights: • We developed a novel firefly luciferase based biosensor to detect apoptosis. • The novel biosensor 233-DnaE-DEVDG was reliable, sensitive and convenient. • 233-DnaE-DEVDG faithfully indicated ESAT-6 family proteins of Mycobacterium tuberculosis induced apoptosis. • EsxA, esxT and esxL in ESAT-6 family proteins induced apoptosis. • Activation of nuclear factor-κB (NF-κB) participated in esxT-induced apoptosis. - Abstract: The activation of caspase-3 is a key surrogate marker for detecting apoptosis. To quantitate caspase-3 activity, we constructed a biosensor comprising a recombinant firefly luciferase containing a caspase-3 cleavage site. When apoptosis was induced, caspase-3 cleavage of the biosensor activated firefly luciferase by a factor greater than 25. The assay conveniently detected apoptosis in real time, indicating that it will facilitate drug discovery. We screened ESAT-6 family proteins of Mycobacterium tuberculosis and found that esxA, esxT and esxL induced apoptosis. Further, activation of nuclear factor-κB (NF-κB) and the NF-κB-regulated genes encoding tumor necrosis factor-α (TNF-α) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) participated in esxT-induced apoptosis. We conclude that this assay is useful for high-throughput screening to identify and characterize proteins and drugs that regulate apoptosis.

  12. Luciferase-Zinc-Finger System for the Rapid Detection of Pathogenic Bacteria.

    Science.gov (United States)

    Shi, Chu; Xu, Qing; Ge, Yue; Jiang, Ling; Huang, He

    2017-08-09

    Rapid and reliable detection of pathogenic bacteria is crucial for food safety control. Here, we present a novel luciferase-zinc finger system for the detection of pathogens that offers rapid and specific profiling. The system, which uses a zinc-finger protein domain to probe zinc finger recognition sites, was designed to bind the amplified conserved regions of 16S rDNA, and the obtained products were detected using a modified luciferase. The luciferase-zinc finger system not only maintained luciferase activity but also allowed the specific detection of different bacterial species, with a sensitivity as low as 10 copies and a linear range from 10 to 10 4 copies per microliter of the specific PCR product. Moreover, the system is robust and rapid, enabling the simultaneous detection of 6 species of bacteria in artificially contaminated samples with excellent accuracy. Thus, we envision that our luciferase-zinc finger system will have far-reaching applications.

  13. High-Throughput Screening of a Luciferase Reporter of Gene Silencing on the Inactive X Chromosome.

    Science.gov (United States)

    Keegan, Alissa; Plath, Kathrin; Damoiseaux, Robert

    2018-01-01

    Assays of luciferase gene activity are a sensitive and quantitative reporter system suited to high-throughput screening. We adapted a luciferase assay to a screening strategy for identifying factors that reactivate epigenetically silenced genes. This epigenetic luciferase reporter is subject to endogenous gene silencing mechanisms on the inactive X chromosome (Xi) in primary mouse cells and thus captures the multilayered nature of chromatin silencing in development. Here, we describe the optimization of an Xi-linked luciferase reactivation assay in 384-well format and adaptation of the assay for high-throughput siRNA and chemical screening. Xi-luciferase reactivation screening has applications in stem cell biology and cancer therapy. We have used the approach described here to identify chromatin-modifying proteins and to identify drug combinations that enhance the gene reactivation activity of the DNA demethylating drug 5-aza-2'-deoxycytidine.

  14. Ozone Depletion Caused by Rocket Engine Emissions: A Fundamental Limit on the Scale and Viability of Space-Based Geoengineering Schemes

    Science.gov (United States)

    Ross, M. N.; Toohey, D.

    2008-12-01

    Emissions from solid and liquid propellant rocket engines reduce global stratospheric ozone levels. Currently ~ one kiloton of payloads are launched into earth orbit annually by the global space industry. Stratospheric ozone depletion from present day launches is a small fraction of the ~ 4% globally averaged ozone loss caused by halogen gases. Thus rocket engine emissions are currently considered a minor, if poorly understood, contributor to ozone depletion. Proposed space-based geoengineering projects designed to mitigate climate change would require order of magnitude increases in the amount of material launched into earth orbit. The increased launches would result in comparable increases in the global ozone depletion caused by rocket emissions. We estimate global ozone loss caused by three space-based geoengineering proposals to mitigate climate change: (1) mirrors, (2) sunshade, and (3) space-based solar power (SSP). The SSP concept does not directly engineer climate, but is touted as a mitigation strategy in that SSP would reduce CO2 emissions. We show that launching the mirrors or sunshade would cause global ozone loss between 2% and 20%. Ozone loss associated with an economically viable SSP system would be at least 0.4% and possibly as large as 3%. It is not clear which, if any, of these levels of ozone loss would be acceptable under the Montreal Protocol. The large uncertainties are mainly caused by a lack of data or validated models regarding liquid propellant rocket engine emissions. Our results offer four main conclusions. (1) The viability of space-based geoengineering schemes could well be undermined by the relatively large ozone depletion that would be caused by the required rocket launches. (2) Analysis of space- based geoengineering schemes should include the difficult tradeoff between the gain of long-term (~ decades) climate control and the loss of short-term (~ years) deep ozone loss. (3) The trade can be properly evaluated only if our

  15. The effect of a non-denaturing detergent and a guanidinium-based inactivation agent on the viability of Ebola virus in mock clinical serum samples.

    Science.gov (United States)

    Burton, J E; Easterbrook, L; Pitman, J; Anderson, D; Roddy, S; Bailey, D; Vipond, R; Bruce, C B; Roberts, A D

    2017-12-01

    The 2014 Ebola outbreak in West Africa required the rapid testing of clinical material for the presence of potentially high titre Ebola virus (EBOV). Safe, fast and effective methods for the inactivation of such clinical samples are required so that rapid diagnostic tests including downstream analysis by RT-qPCR or nucleotide sequencing can be carried out. One of the most commonly used guanidinium - based denaturing agents, AVL (Qiagen) has been shown to fully inactivate EBOV once ethanol is added, however this is not compatible with the use of automated nucleic acid extraction systems. Additional inactivation agents need to be identified that can be used in automated systems. A candidate inactivation agent is Triton X-100, a non-denaturing detergent that is frequently used in clinical nucleic acid extraction procedures and has previously been used for inactivation of EBOV. In this study the effect of 0.1% and 1.0% Triton X-100 (final concentration 0.08% and 0.8% respectively) alone and in combination with AVL on the viability of EBOV (10 6 TCID 50 /ml) spiked into commercially available pooled negative human serum was tested. The presence of viable EBOV in the treated samples was assessed by carrying out three serial passages of the samples in Vero E6 cells (37°C, 5% CO 2 , 1 week for each passage). At the end of each passage the cells were observed for evidence of cytopathic effect and samples were taken for rRT-PCR analysis for the presence of EBOV RNA. Before cell culture cytotoxic components of AVL and Triton X-100 were removed from the samples using size exclusion spin column technology or a hydrophobic adsorbent resin. The results of this study showed that EBOV spiked into human serum was not fully inactivated when treated with either 0.1% (v/v) Triton X-100 for 10 mins or 1.0% (v/v) Triton X-100 for 20 mins (final concentrations 0.08% and 0.8% Triton X-100 respectively). AVL alone also did not consistently provide complete inactivation. Samples treated

  16. Inefficiency in macromolecular transport of SCS-based microcapsules affects viability of primary human mesenchymal stem cells but not of immortalized cells

    DEFF Research Database (Denmark)

    Sanz-Nogués, Clara; Horan, Jason; Thompson, Kerry

    2015-01-01

    mesenchymal stem cells (hMSCs). Human MSCs are of interest in regenerative medicine applications due to pro-angiogenic, anti-inflammatory and immunomodulatory properties, which result from paracrine effects of this cell type. In the present work we have encapsulated primary hMSCs and hMSC-TERT immortalized...... nutrients and had a more detrimental effect on the viability of primary cell cultures compared to cell lines and immortalized cells. This article is protected by copyright. All rights reserved....

  17. Luciferase-Specific Coelenterazine Analogues for Optical Contamination-Free Bioassays.

    Science.gov (United States)

    Nishihara, Ryo; Abe, Masahiro; Nishiyama, Shigeru; Citterio, Daniel; Suzuki, Koji; Kim, Sung Bae

    2017-04-19

    Spectral overlaps among the multiple optical readouts commonly cause optical contamination in fluorescence and bioluminescence. To tackle this issue, we created five-different lineages of coelenterazine (CTZ) analogues designed to selectively illuminate a specific luciferase with unique luciferase selectivity. In the attempt, we found that CTZ analogues with ethynyl or styryl groups display dramatically biased bioluminescence to specific luciferases and pHs by modifying the functional groups at the C-2 and C-6 positions of the imidazopyradinone backbone of CTZ. The optical contamination-free feature was exemplified with the luciferase-specific CTZ analogues, which illuminated anti-estrogenic and rapamycin activities in a mixture of optical probes. This unique bioluminescence platform has great potential for specific and high throughput imaging of multiple optical readouts in bioassays without optical contamination.

  18. Luciferase genes cloned from the unculturable luminous bacteroid symbiont of the Caribbean flashlight fish, Kryptophanaron alfredi.

    Science.gov (United States)

    Haygood, M G; Cohn, D H

    1986-01-01

    Light organs of anomalopid (flashlight) fish contain luminous bacteroids that have never been cultured and, consequently, have been difficult to study. We have characterized the luciferase (lux) region of DNA extracted from light organs of the Caribbean flashlight fish Kryptophanaron alfredi by hybridization of cloned Vibrio harveyi lux genes to restriction-endonuclease-digested, light organ DNA. Comparison of the hybridization pattern of light organ DNA with that of DNA of a putative symbiotic isolate provides a method for identifying the authentic luminous symbiont regardless of its luminescence, and was used to reject one such isolate. Light organ DNA was further used to construct a cosmid clone bank and the luciferase genes were isolated. Unlike other bacterial luciferase genes, the genes were not expressed in Escherichia coli. When placed under the control of the E. coli trp promoter, the genes were transcribed but no luciferase was detected, suggesting a posttranscriptional block to expression.

  19. Probing the emitter site of Renilla luciferase using small organic molecules; an attempt to understand the molecular architecture of the emitter site.

    Science.gov (United States)

    Salehi, Farajollah; Emamzadeh, Rahman; Nazari, Mahboobeh; Rasa, Seyed Mohammad Mahdi

    2016-12-01

    Renilla luciferase is a sensitive enzyme and has wide applications in biotechnology such as drug screening. Previous studies have tried to show the catalytic residues, nevertheless, the accurate architecture and molecular behavior of its emitter site remains uncharacterized. In this study, the activity of Renilla luciferase, in the presence of two small organic molecules including dimethyl sulfoxide (DMSO) and isopropanol was considered and the structure was studied by circular dichroism (CD) and fluorescence spectroscopy. Moreover, the interaction of small organic molecules with the Renilla luciferase was studied using molecular dynamics simulations. Kinetics studies showed that at low concentration of DMSO (16.6-66mM) and isopropanol (19.3-76mM) the K m changed and a competitive inhibition pattern was observed. Moreover, spectroscopy studies reveled that the changes of activity of Renilla luciferase in the presence of low concentrations of small organic molecules was not associated with structural collapse or severe changes in the enzyme conformation. Molecular dynamics simulations indicated that DMSO and isopropanol, as probing molecules, were both able to bind to the emitter site and remained with the residues of the emitter site. Based on the probing data, the architecture of the emitter site in the "non-binding" model was proposed. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Characterization of CG6178 gene product with high sequence similarity to firefly luciferase in Drosophila melanogaster.

    Science.gov (United States)

    Oba, Yuichi; Ojika, Makoto; Inouye, Satoshi

    2004-03-31

    This is the first identification of a long-chain fatty acyl-CoA synthetase in Drosophila by enzymatic characterization. The gene product of CG6178 (CG6178) in Drosophila melanogaster genome, which has a high sequence similarity to firefly luciferase, has been expressed and characterized. CG6178 showed long-chain fatty acyl-CoA synthetic activity in the presence of ATP, CoA and Mg(2+), suggesting a fatty acyl adenylate is an intermediate. Recently, it was revealed that firefly luciferase has two catalytic functions, monooxygenase (luciferase) and AMP-mediated CoA ligase (fatty acyl-CoA synthetase). However, unlike firefly luciferase, CG6178 did not show luminescence activity in the presence of firefly luciferin, ATP, CoA and Mg(2+). The enzymatic properties of CG6178 including substrate specificity, pH dependency and optimal temperature were close to those of firefly luciferase and rat fatty acyl-CoA synthetase. Further, phylogenic analyses strongly suggest that the firefly luciferase gene may have evolved from a fatty acyl-CoA synthetase gene as a common ancestral gene.

  1. Cloning and characterization of luciferase from a Fijian luminous click beetle.

    Science.gov (United States)

    Mitani, Yasuo; Futahashi, Ryo; Niwa, Kazuki; Ohba, Nobuyoshi; Ohmiya, Yoshihiro

    2013-01-01

    Luminous click beetle is distributed almost exclusively in Central and South America with a single genus in Melanesia. Among these click beetles, the description of Melanesian species has been fragmentary, and its luciferase gene and phylogenetic relation to other click beetles still remain uncertain. We collected a living luminous click beetle, Photophorus jansonii in Fiji. It emits green-yellow light from two spots on the pronotum and has no ventral luminous organ. Here, we cloned a luciferase gene from this insect by RT-PCR. The deduced amino acid sequence showed high identity of ~85% to the luciferases derived from other click beetle species. The luciferase of the Fijian click beetle was produced as a recombinant protein to characterize its biochemical properties. The Km for D-luciferin and ATP were 173 and 270 μm, respectively. The luciferase was pH-insensitive and the spectrum measured at pH 8.0 showed a peak at 559 nm, which was in the range of green-yellow light as seen in the luminous spot of the living Fijian click beetle. The Fijian click beetle luciferase was assigned to the Elateridae clade by a phylogenetic analysis, but it made a clearly different branch from Pyrophorus group examined in this study. © 2013 The American Society of Photobiology.

  2. Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase

    International Nuclear Information System (INIS)

    Yoshida, Wataru; Kezuka, Aki; Murakami, Yoshiyuki; Lee, Jinhee; Abe, Koichi; Motoki, Hiroaki; Matsuo, Takafumi; Shimura, Nobuaki; Noda, Mamoru; Igimi, Shizunobu; Ikebukuro, Kazunori

    2013-01-01

    Graphical abstract: -- Highlights: •Zif268 fused to luciferase was used for E. coli O157, Salmonella and coliform detection. •Artificial zinc finger protein fused to luciferase was constructed for Norovirus detection. •An analyzer that automatically detects PCR products by zinc finger protein fused to luciferase was developed. •Target pathogens were specifically detected by the automatic analyzer with zinc finger protein fused to luciferase. -- Abstract: An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268–luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF–luciferase fusion protein. By means of the automatic analyzer with ZF–luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0 × 10 to 1.0 × 10 6 copies

  3. Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase

    Energy Technology Data Exchange (ETDEWEB)

    Yoshida, Wataru; Kezuka, Aki; Murakami, Yoshiyuki; Lee, Jinhee; Abe, Koichi [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 (Japan); Motoki, Hiroaki; Matsuo, Takafumi; Shimura, Nobuaki [System Instruments Co., Ltd., 776-2 Komiya-cho, Hachioji, Tokyo 192-0031 (Japan); Noda, Mamoru; Igimi, Shizunobu [Division of Biomedical Food Research, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501 (Japan); Ikebukuro, Kazunori, E-mail: ikebu@cc.tuat.ac.jp [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 (Japan)

    2013-11-01

    Graphical abstract: -- Highlights: •Zif268 fused to luciferase was used for E. coli O157, Salmonella and coliform detection. •Artificial zinc finger protein fused to luciferase was constructed for Norovirus detection. •An analyzer that automatically detects PCR products by zinc finger protein fused to luciferase was developed. •Target pathogens were specifically detected by the automatic analyzer with zinc finger protein fused to luciferase. -- Abstract: An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268–luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF–luciferase fusion protein. By means of the automatic analyzer with ZF–luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0 × 10 to 1.0 × 10{sup 6} copies.

  4. Algae viability over time in a ballast water sample

    Science.gov (United States)

    Gollasch, Stephan; David, Matej

    2018-03-01

    The biology of vessels' ballast water needs to be analysed for several reasons, one of these being performance tests of ballast water management systems. This analysis includes a viability assessment of phytoplankton. To overcome logistical problems to get algae sample processing gear on board of a vessel to document algae viability, samples may be transported to land-based laboratories. Concerns were raised how the storage conditions of the sample may impact algae viability over time and what the most appropriate storage conditions were. Here we answer these questions with a long-term algae viability study with daily sample analysis using Pulse-Amplitude Modulated (PAM) fluorometry. The sample was analysed over 79 days. We tested different storage conditions: fridge and room temperature with and without light. It seems that during the first two weeks of the experiment the viability remains almost unchanged with a slight downwards trend. In the continuing period, before the sample was split, a slightly stronger downwards viability trend was observed, which occurred at a similar rate towards the end of the experiment. After the sample was split, the strongest viability reduction was measured for the sample stored without light at room temperature. We concluded that the storage conditions, especially regarding temperature and light exposure, have a stronger impact on algae viability compared to the storage duration and that inappropriate storage conditions reduce algal viability. A sample storage time of up to two weeks in a dark and cool environment has little influence on the organism viability. This indicates that a two week time duration between sample taking on board a vessel and the viability measurement in a land-based laboratory may not be very critical.

  5. Skin graft viability test

    International Nuclear Information System (INIS)

    Wahner, H.W.; Robertson, J.S.

    1983-01-01

    With respect to skin pedicles (tubular pedicle or direct flap), an estimation of the blood supply is of great importance in making a decision as to when to perform the separation from the original blood supply. This decision is based on empiric observation of the normal time of healing and varies greatly with the site and the concepts of the individual surgery. A number of methods have been proposed for testing the circulation of pedicle skin flaps or tubes, and these methods all seem more complicated and less accurate than the isotope method that is described in this chapter

  6. Application of firefly luciferase assay for adenosine triphosphate (ATP) to antimicrobial drug sensitivity testing

    Science.gov (United States)

    Picciolo, G. L.; Tuttle, S. A.; Schrock, C. G.; Deming, J. W.; Barza, M. J.; Wienstein, L.; Chappelle, E. W.

    1977-01-01

    The development of a rapid method for determining microbial susceptibilities to antibiotics using the firefly luciferase assay for adenosine triphosphate (ATP) is documented. The reduction of bacterial ATP by an antimicrobial agent was determined to be a valid measure of drug effect in most cases. The effect of 12 antibiotics on 8 different bacterial species gave a 94 percent correlation with the standard Kirby-Buer-Agar disc diffusion method. A 93 percent correlation was obtained when the ATP assay method was applied directly to 50 urine specimens from patients with urinary tract infections. Urine samples were centrifuged first to that bacterial pellets could be suspended in broth. No primary isolation or subculturing was required. Mixed cultures in which one species was predominant gave accurate results for the most abundant organism. Since the method is based on an increase in bacterial ATP with time, the presence of leukocytes did not interfere with the interpretation of results. Both the incubation procedure and the ATP assays are compatible with automation.

  7. Application of the dual-luciferase reporter assay to the analysis of promoter activity in Zebrafish embryos

    Directory of Open Access Journals (Sweden)

    Mulero Victoriano

    2008-10-01

    Full Text Available Abstract Background The dual-luciferase assay has been widely used in cell lines to determine rapidly but accurately the activity of a given promoter. Although this strategy has proved very useful, it does not allow the promoter and gene function to be analyzed in the context of the whole organism. Results Here, we present a rapid and sensitive assay based on the classical dual-luciferase reporter technique which can be used as a new tool to characterize the minimum promoter region of a gene as well as the in vivo response of inducible promoters to different stimuli. We illustrate the usefulness of this system for studying both constitutive (telomerase and inducible (NF-κB-dependent promoters. The flexibility of this assay is demonstrated by induction of the NF-κB-dependent promoters using simultaneous microinjection of different pathogen-associated molecular patterns as well as with the use of morpholino-gene mediated knockdown. Conclusion This assay has several advantages compared with the classical in vitro (cell lines and in vivo (transgenic mice approaches. Among others, the assay allows a rapid and quantitative measurement of the effects of particular genes or drugs in a given promoter in the context of a whole organism and it can also be used in high throughput screening experiments.

  8. The cAMP-dependent protein kinase inhibitor H-89 attenuates the bioluminescence signal produced by Renilla Luciferase.

    Directory of Open Access Journals (Sweden)

    Katie J Herbst

    2009-05-01

    Full Text Available Investigations into the regulation and functional roles of kinases such as cAMP-dependent protein kinase (PKA increasingly rely on cellular assays. Currently, there are a number of bioluminescence-based assays, for example reporter gene assays, that allow the study of the regulation, activity, and functional effects of PKA in the cellular context. Additionally there are continuing efforts to engineer improved biosensors that are capable of detecting real-time PKA signaling dynamics in cells. These cell-based assays are often utilized to test the involvement of PKA-dependent processes by using H-89, a reversible competitive inhibitor of PKA.We present here data to show that H-89, in addition to being a competitive PKA inhibitor, attenuates the bioluminescence signal produced by Renilla luciferase (RLuc variants in a population of cells and also in single cells. Using 10 microM of luciferase substrate and 10 microM H-89, we observed that the signal from RLuc and RLuc8, an eight-point mutation variant of RLuc, in cells was reduced to 50% (+/-15% and 54% (+/-14% of controls exposed to the vehicle alone, respectively. In vitro, we showed that H-89 decreased the RLuc8 bioluminescence signal but did not compete with coelenterazine-h for the RLuc8 active site, and also did not affect the activity of Firefly luciferase. By contrast, another competitive inhibitor of PKA, KT5720, did not affect the activity of RLuc8.The identification and characterization of the adverse effect of H-89 on RLuc signal will help deconvolute data previously generated from RLuc-based assays looking at the functional effects of PKA signaling. In addition, for the current application and future development of bioluminscence assays, KT5720 is identified as a more suitable PKA inhibitor to be used in conjunction with RLuc-based assays. These principal findings also provide an important lesson to fully consider all of the potential effects of experimental conditions on a cell-based

  9. Cell banking for regulatory T cell-based therapy: strategies to overcome the impact of cryopreservation on the Treg viability and phenotype.

    Science.gov (United States)

    Gołąb, Karolina; Grose, Randall; Placencia, Veronica; Wickrema, Amittha; Solomina, Julia; Tibudan, Martin; Konsur, Evelyn; Ciepły, Kamil; Marek-Trzonkowska, Natalia; Trzonkowski, Piotr; Millis, J Michael; Fung, John; Witkowski, Piotr

    2018-02-09

    The first clinical trials with adoptive Treg therapy have shown safety and potential efficacy. Feasibility of such therapy could be improved if cells are cryopreserved and stored until optimal timing for infusion. Herein, we report the evaluation of two cell-banking strategies for Treg therapy: 1) cryopreservation of CD4 + cells for subsequent Treg isolation/expansion and 2) cryopreservation of ex-vivo expanded Tregs (CD4 + CD25 hi CD127 lo/- cells). First, we checked how cryopreservation affects cell viability and Treg markers expression. Then, we performed Treg isolation/expansion with the final products release testing. We observed substantial decrease in cell number recovery after thawing and overnight culture. This observation might be explained by the high percentage of necrotic and apoptotic cells found just after thawing. Furthermore, we noticed fluctuations in percentage of CD4 + CD25 hi CD127 - and CD4 + FoxP3 + cells obtained from cryopreserved CD4 + as well as Treg cells. However, after re-stimulation Tregs expanded well, presented a stable phenotype and fulfilled the release criteria at the end of expansions. Cryopreservation of CD4 + cells for subsequent Treg isolation/expansion and cryopreservation of expanded Tregs with re-stimulation and expansion after thawing, are promising solutions to overcome detrimental effects of cryopreservation. Both of these cell-banking strategies for Treg therapy can be applied when designing new clinical trials.

  10. Viability Test Device for anisakid nematodes

    Directory of Open Access Journals (Sweden)

    Michael Kroeger

    2018-03-01

    Full Text Available Up to now the visual inspection of mobility of isolated anisakid larvae serves as a measure of viability and possible risk of infection. This paper presents a new method to rule out unreliability – caused by the temporary immobility of the larvae and by the human uncertainty factor of visual observation. By means of a Near infrared (NIR imaging method, elastic curvature energies and geometric shape parameters were determined from contours, and used as a measure of viability. It was based on the modelling of larvae as a cylindrical membrane system. The interaction between curvatures, contraction of the longitudinal muscles, and inner pressure enabled the derivation of viability from stationary form data. From series of spectrally signed images within a narrow wavelength range, curvature data of the larvae were determined. Possible mobility of larvae was taken into account in statistical error variables. Experiments on individual living larvae, long-term observations of Anisakis larvae, and comparative studies of the staining method and the VTD measurements of larvae from the tissue of products confirmed the effectiveness of this method. The VTD differentiated clearly between live and dead nematode larvae isolated from marinated, deep-frozen and salted products. The VTD has been proven as excellent method to detect living anisakid nematode larvae in fishery products and is seen as useful tool for fish processing industry and control authorities. Keywords: Biophysics

  11. A viability analysis for a stock/price model

    Science.gov (United States)

    Jerry, Chakib; Raissi, Nadia

    2012-09-01

    We examine the conditions for the sustainability of a stock/price system based on the use of a marine renewable resource. Instead of studying the environmental and economic interactions in terms of optimal control, we focus on the viability of the system. These viability/crisis situations are defined by a set of economic state constraints. This constraints combine a guaranteed consumption and a minimum income for fishermen. Using the mathematical concept of viability kernel, we reveal that with only economics constraints we guarantee a perennial stock/price system.

  12. Bioluminescence of beetle luciferases with 6'-amino-D-luciferin analogues reveals excited keto-oxyluciferin as the emitter and phenolate/luciferin binding site interactions modulate bioluminescence colors.

    Science.gov (United States)

    Viviani, Vadim R; Neves, Deimison Rodrigues; Amaral, Danilo Trabuco; Prado, Rogilene A; Matsuhashi, Takuto; Hirano, Takashi

    2014-08-19

    Beetle luciferases produce different bioluminescence colors from green to red using the same d-luciferin substrate. Despite many studies of the mechanisms and structural determinants of bioluminescence colors with firefly luciferases, the identity of the emitters and the specific active site interactions responsible for bioluminescence color modulation remain elusive. To address these questions, we analyzed the bioluminescence spectra with 6'-amino-D-luciferin (aminoluciferin) and its 5,5-dimethyl analogue using a set of recombinant beetle luciferases that naturally elicit different colors and different pH sensitivities (pH-sensitive, Amydetes vivianii λmax=538 nm, Macrolampis sp2 λmax=564 nm; pH-insensitive, Phrixotrix hirtus λmax=623 nm, Phrixotrix vivianii λmax=546 nm, and Pyrearinus termitilluminans λmax=534 nm), a luciferase-like enzyme (Tenebrionidae, Zophobas morio λmax=613 nm), and mutants of C311 (S314). The green-yellow-emitting luciferases display red-shifted bioluminescence spectra with aminoluciferin in relation to those with D-luciferin, whereas the red-emitting luciferases displayed blue-shifted spectra. Bioluminescence spectra with 5,5-dimethylaminoluciferin, in which enolization is blocked, were almost identical to those of aminoluciferin. Fluorescence probing using 2-(4-toluidino)naphthalene-6-sulfonate and inference with aminoluciferin confirm that the luciferin binding site of the red-shifted luciferases is more polar than in the case of the green-yellow-emitting luciferases. Altogether, the results show that the keto form of excited oxyluciferin is the emitter in beetle bioluminescence and that bioluminescence colors are essentially modulated by interactions of the 6'-hydroxy group of oxyluciferin and basic moieties under the influence of the microenvironment polarity of the active site: a strong interaction between a base moiety and oxyluciferin phenol in a hydrophobic microenvironment promotes green-yellow emission, whereas a more polar

  13. The Evolution of the Bacterial Luciferase Gene Cassette (lux as a Real-Time Bioreporter

    Directory of Open Access Journals (Sweden)

    Gary Sayler

    2012-01-01

    Full Text Available The bacterial luciferase gene cassette (lux is unique among bioluminescent bioreporter systems due to its ability to synthesize and/or scavenge all of the substrate compounds required for its production of light. As a result, the lux system has the unique ability to autonomously produce a luminescent signal, either continuously or in response to the presence of a specific trigger, across a wide array of organismal hosts. While originally employed extensively as a bacterial bioreporter system for the detection of specific chemical signals in environmental samples, the use of lux as a bioreporter technology has continuously expanded over the last 30 years to include expression in eukaryotic cells such as Saccharomyces cerevisiae and even human cell lines as well. Under these conditions, the lux system has been developed for use as a biomedical detection tool for toxicity screening and visualization of tumors in small animal models. As the technologies for lux signal detection continue to improve, it is poised to become one of the first fully implantable detection systems for intra-organismal optical detection through direct marriage to an implantable photon-detecting digital chip. This review presents the basic biochemical background that allows the lux system to continuously autobioluminesce and highlights the important milestones in the use of lux-based bioreporters as they have evolved from chemical detection platforms in prokaryotic bacteria to rodent-based tumorigenesis study targets. In addition, the future of lux imaging using integrated circuit microluminometry to image directly within a living host in real-time will be introduced and its role in the development of dose/response therapeutic systems will be highlighted.

  14. Viability of infrared FEL facilities

    International Nuclear Information System (INIS)

    Schwettman, H.A.

    2004-01-01

    Infrared FELs have broken important ground in optical science in the past decade. The rapid development of optical parametric amplifiers and oscillators, and THz sources, however, has changed the competitive landscape and compelled FEL facilities to identify and exploit their unique advantages. The viability of infrared FEL facilities depends on targeting unique world-class science and providing adequate experimental beam time at competitive costs

  15. A Dual Luciferase Reporter System for B. burgdorferi Measures Transcriptional Activity during Tick-Pathogen Interactions

    Directory of Open Access Journals (Sweden)

    Philip P. Adams

    2017-05-01

    Full Text Available Knowledge of the transcriptional responses of vector-borne pathogens at the vector-pathogen interface is critical for understanding disease transmission. Borrelia (Borreliella burgdorferi, the causative agent of Lyme disease in the United States, is transmitted by the bite of infected Ixodes sp. ticks. It is known that B. burgdorferi has altered patterns of gene expression during tick acquisition, persistence and transmission. Recently, we and others have discovered in vitro expression of RNAs found internal, overlapping, and antisense to annotated open reading frames in the B. burgdorferi genome. However, there is a lack of molecular genetic tools for B. burgdorferi for quantitative, strand-specific, comparative analysis of these transcripts in distinct environments such as the arthropod vector. To address this need, we have developed a dual luciferase reporter system to quantify B. burgdorferi promoter activities in a strand-specific manner. We demonstrate that constitutive expression of a B. burgdorferi codon-optimized Renilla reniformis luciferase gene (rlucBb allows normalization of the activity of a promoter of interest when fused to the B. burgdorferi codon-optimized Photinus pyralis luciferase gene (flucBb on the same plasmid. Using the well characterized, differentially regulated, promoters for flagellin (flaBp, outer surface protein A (ospAp and outer surface protein C (ospCp, we document the efficacy of the dual luciferase system for quantitation of promoter activities during in vitro growth and in infected ticks. Cumulatively, the dual luciferase method outlined herein is the first dual reporter system for B. burgdorferi, providing a novel and highly versatile approach for strand-specific molecular genetic analyses.

  16. Third-generation Ah receptor-responsive luciferase reporter plasmids: amplification of dioxin-responsive elements dramatically increases CALUX bioassay sensitivity and responsiveness.

    Science.gov (United States)

    He, Guochun; Tsutsumi, Tomoaki; Zhao, Bin; Baston, David S; Zhao, Jing; Heath-Pagliuso, Sharon; Denison, Michael S

    2011-10-01

    2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related dioxin-like chemicals are widespread and persistent environmental contaminants that produce diverse toxic and biological effects through their ability to bind to and activate the Ah receptor (AhR) and AhR-dependent gene expression. The chemically activated luciferase expression (CALUX) system is an AhR-responsive recombinant luciferase reporter gene-based cell bioassay that has been used in combination with chemical extraction and cleanup methods for the relatively rapid and inexpensive detection and relative quantitation of dioxin and dioxin-like chemicals in a wide variety of sample matrices. Although the CALUX bioassay has been validated and used extensively for screening purposes, it has some limitations when screening samples with very low levels of dioxin-like chemicals or when there is only a small amount of sample matrix for analysis. Here, we describe the development of third-generation (G3) CALUX plasmids with increased numbers of dioxin-responsive elements, and stable transfection of these new plasmids into mouse hepatoma (Hepa1c1c7) cells has produced novel amplified G3 CALUX cell bioassays that respond to TCDD with a dramatically increased magnitude of luciferase induction and significantly lower minimal detection limit than existing CALUX-type cell lines. The new G3 CALUX cell lines provide a highly responsive and sensitive bioassay system for the detection and relative quantitation of very low levels of dioxin-like chemicals in sample extracts.

  17. Abundance, viability and culturability of Antarctic bacteria

    Digital Repository Service at National Institute of Oceanography (India)

    LokaBharathi, P.A.; DeSouza, M.J.B.D.; Nair, S.; Chandramohan, D.

    The viability of total number of bacteria decide the mineralisation rate in any ecosystem and ultimately the fertility of the region. This study aims at establishing the extent of viability in the standing stock of the Antarctic bacterial population...

  18. Tychastic measure of viability risk

    CERN Document Server

    Aubin, Jean-Pierre; Dordan, Olivier

    2014-01-01

    This book presents a forecasting mechanism of the price intervals for deriving the SCR (solvency capital requirement) eradicating the risk during the exercise period on one hand, and measuring the risk by computing the hedging exit time function associating with smaller investments the date until which the value of the portfolio hedges the liabilities on the other. This information, summarized under the term “tychastic viability measure of risk” is an evolutionary alternative to statistical measures, when dealing with evolutions under uncertainty. The book is written by experts in the field and the target audience primarily comprises research experts and practitioners.

  19. Combined image guided monitoring the pharmacokinetics of rapamycin loaded human serum albumin nanoparticles with a split luciferase reporter

    Science.gov (United States)

    Wang, Fu; Yang, Kai; Wang, Zhe; Ma, Ying; Gutkind, J. Silvio; Hida, Naoki; Niu, Gang; Tian, Jie

    2016-02-01

    Imaging guided techniques have been increasingly employed to investigate the pharmacokinetics (PK) and biodistribution of nanoparticle based drug delivery systems. In most cases, however, the PK profiles of drugs could vary significantly from those of drug delivery carriers upon administration in the blood circulation, which complicates the interpretation of image findings. Herein we applied a genetically encoded luciferase reporter in conjunction with near infrared (NIR) fluorophores to investigate the respective PK profiles of a drug and its carrier in a biodegradable drug delivery system. In this system, a prototype hydrophobic agent, rapamycin (Rapa), was encapsulated into human serum albumin (HSA) to form HSA Rapa nanoparticles, which were then labeled with Cy5 fluorophore to facilitate the fluorescence imaging of HSA carrier. Meanwhile, we employed transgenetic HN12 cells that were modified with a split luciferase reporter, whose bioluminescence function is regulated by Rapa, to reflect the PK profile of the encapsulated agent. It was interesting to discover that there existed an obvious inconsistency of PK behaviors between HSA carrier and rapamycin in vitro and in vivo through near infrared fluorescence imaging (NIFRI) and bioluminescence imaging (BLI) after treatment with Cy5 labeled HSA Rapa. Nevertheless, HSA Rapa nanoparticles manifested favorable in vivo PK and tumor suppression efficacy in a follow-up therapeutic study. The developed strategy of combining a molecular reporter and a fluorophore in this study could be extended to other drug delivery systems to provide profound insights for non-invasive real-time evaluation of PK profiles of drug-loaded nanoparticles in pre-clinical studies.Imaging guided techniques have been increasingly employed to investigate the pharmacokinetics (PK) and biodistribution of nanoparticle based drug delivery systems. In most cases, however, the PK profiles of drugs could vary significantly from those of drug delivery

  20. Step-wise addition of disulfide bridge in firefly luciferase controls color shift through a flexible loop: a thermodynamic perspective.

    Science.gov (United States)

    Nazari, Mahboobeh; Hosseinkhani, Saman; Hassani, Leila

    2013-02-01

    Multi-color bioluminescence is developed using the introduction of single/double disulfide bridges in firefly luciferase. The bioluminescence reaction, which uses luciferin, Mg(2+)-ATP and molecular oxygen to yield an electronically excited oxyluciferin, is carried out by the luciferase and emits visible light. The bioluminescence color of firefly luciferases is determined by the luciferase sequence and assay conditions. It has been proposed that the stability of a protein may increase through the introduction of a disulfide bridge that decreases the configurational entropy of unfolding. Single and double disulfide bridges are introduced into Photinus pyralis firefly luciferase to make separate mutant enzymes with a single/double bridge (C(81)-A(105)C, L(306)C-L(309)C, P(451)C-V(469)C; C(81)-A(105)C/P(451)C-V(469)C, and A(296)C-A(326)C/P(451)C-V(469)C). By introduction of disulfide bridges using site-directed mutagenesis in Photinus pyralis luciferase the color of emitted light was changed to red or kept in different extents. The bioluminescence color shift occurred with displacement of a critical loop in the luciferase structure without any change in green emitter mutants. Thermodynamic analysis revealed that among mutants, L(306)C-L(309)C shows a remarkable stability against urea denaturation and also a considerable increase in kinetic stability and a clear shift in bioluminescence spectra towards red.

  1. A mouse model of pulmonary metastasis from spontaneous osteosarcoma monitored in vivo by Luciferase imaging.

    Directory of Open Access Journals (Sweden)

    Silvia Miretti

    Full Text Available BACKGROUND: Osteosarcoma (OSA is lethal when metastatic after chemotherapy and/or surgical treatment. Thus animal models are necessary to study the OSA metastatic spread and to validate novel therapies able to control the systemic disease. We report the development of a syngeneic (Balb/c murine OSA model, using a cell line derived from a spontaneous murine tumor. METHODOLOGY: The tumorigenic and metastatic ability of OSA cell lines were assayed after orthotopic injection in mice distal femur. Expression profiling was carried out to characterize the parental and metastatic cell lines. Cells from metastases were propagated and engineered to express Luciferase, in order to follow metastases in vivo. PRINCIPAL FINDINGS: Luciferase bioluminescence allowed to monitor the primary tumor growth and revealed the appearance of spontaneous pulmonary metastases. In vivo assays showed that metastasis is a stable property of metastatic OSA cell lines after both propagation in culture and luciferase trasduction. When compared to parental cell line, both unmodified and genetically marked metastatic cells, showed comparable and stable differential expression of the enpp4, pfn2 and prkcd genes, already associated to the metastatic phenotype in human cancer. CONCLUSIONS: This OSA animal model faithfully recapitulates some of the most important features of the human malignancy, such as lung metastatization. Moreover, the non-invasive imaging allows monitoring the tumor progression in living mice. A great asset of this model is the metastatic phenotype, which is a stable property, not modifiable after genetic manipulation.

  2. A transgenic Plasmodium falciparum NF54 strain that expresses GFP-luciferase throughout the parasite life cycle.

    Science.gov (United States)

    Vaughan, Ashley M; Mikolajczak, Sebastian A; Camargo, Nelly; Lakshmanan, Viswanathan; Kennedy, Mark; Lindner, Scott E; Miller, Jessica L; Hume, Jen C C; Kappe, Stefan H I

    2012-12-01

    Plasmodium falciparum is the pathogenic agent of the most lethal of human malarias. Transgenic P. falciparum parasites expressing luciferase have been created to study drug interventions of both asexual and sexual blood stages but luciferase-expressing mosquito stage and liver stage parasites have not been created which has prevented the easy quantification of mosquito stage development (e.g. for transmission blocking interventions) and liver stage development (for interventions that prevent infection). To overcome this obstacle, we have created a transgenic P. falciparum NF54 parasite that expresses a GFP-luciferase transgene throughout the life cycle. Luciferase expression is robust and measurable at all life cycle stages, including midgut oocyst, salivary gland sporozoites and liver stages, where in vivo development is easily measurable using humanized mouse infections in conjunction with an in vivo imaging system. This parasite reporter strain will accelerate testing of interventions against pre-erythrocytic life cycle stages. Copyright © 2012 Elsevier B.V. All rights reserved.

  3. A Classification Method for Seed Viability Assessment with Infrared Thermography

    Directory of Open Access Journals (Sweden)

    Sen Men

    2017-04-01

    Full Text Available This paper presents a viability assessment method for Pisum sativum L. seeds based on the infrared thermography technique. In this work, different artificial treatments were conducted to prepare seeds samples with different viability. Thermal images and visible images were recorded every five minutes during the standard five day germination test. After the test, the root length of each sample was measured, which can be used as the viability index of that seed. Each individual seed area in the visible images was segmented with an edge detection method, and the average temperature of the corresponding area in the infrared images was calculated as the representative temperature for this seed at that time. The temperature curve of each seed during germination was plotted. Thirteen characteristic parameters extracted from the temperature curve were analyzed to show the difference of the temperature fluctuations between the seeds samples with different viability. With above parameters, support vector machine (SVM was used to classify the seed samples into three categories: viable, aged and dead according to the root length, the classification accuracy rate was 95%. On this basis, with the temperature data of only the first three hours during the germination, another SVM model was proposed to classify the seed samples, and the accuracy rate was about 91.67%. From these experimental results, it can be seen that infrared thermography can be applied for the prediction of seed viability, based on the SVM algorithm.

  4. Population viability analysis on domestic horse breeds (Equus caballus)

    DEFF Research Database (Denmark)

    Thirstrup, Janne Pia; Bach, Lars; Loeschcke, Volker

    2009-01-01

    simulation package was used for the population viability analysis. First, we investigated the future viability of these breeds based on present demographic and environmental parameters. Second, a sensitivity analysis revealed the most important variables for the viability of these breeds. Third, we examined...... concerning reproduction of the mares had the greatest impact, with the number of mares actively breeding being the most influential on the population forecasts. The results suggest that closing the Knabstrupper studbooks can be done only if increasing the number of mares actively breeding counteracts...... the loss of genetic variation attributable to such a management strategy. It is recommended, based on these results, that the number of Frederiksborg and Knabstrupper mares actively breeding must be increased to approximately 30% in the 2 breeds that are presently using only 13%, while leaving the third...

  5. Relative quantification of protein-protein interactions using a dual luciferase reporter pull-down assay system.

    Directory of Open Access Journals (Sweden)

    Shuaizheng Jia

    Full Text Available The identification and quantitative analysis of protein-protein interactions are essential to the functional characterization of proteins in the post-proteomics era. The methods currently available are generally time-consuming, technically complicated, insensitive and/or semi-quantitative. The lack of simple, sensitive approaches to precisely quantify protein-protein interactions still prevents our understanding of the functions of many proteins. Here, we develop a novel dual luciferase reporter pull-down assay by combining a biotinylated Firefly luciferase pull-down assay with a dual luciferase reporter assay. The biotinylated Firefly luciferase-tagged protein enables rapid and efficient isolation of a putative Renilla luciferase-tagged binding protein from a relatively small amount of sample. Both of these proteins can be quantitatively detected using the dual luciferase reporter assay system. Protein-protein interactions, including Fos-Jun located in the nucleus; MAVS-TRAF3 in cytoplasm; inducible IRF3 dimerization; viral protein-regulated interactions, such as MAVS-MAVS and MAVS-TRAF3; IRF3 dimerization; and protein interaction domain mapping, are studied using this novel assay system. Herein, we demonstrate that this dual luciferase reporter pull-down assay enables the quantification of the relative amounts of interacting proteins that bind to streptavidin-coupled beads for protein purification. This study provides a simple, rapid, sensitive, and efficient approach to identify and quantify relative protein-protein interactions. Importantly, the dual luciferase reporter pull-down method will facilitate the functional determination of proteins.

  6. Identification of compounds with anti-proliferative activity against Trypanosoma brucei brucei strain 427 by a whole cell viability based HTS campaign.

    Directory of Open Access Journals (Sweden)

    Melissa L Sykes

    Full Text Available Human African Trypanosomiasis (HAT is caused by two trypanosome sub-species, Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense. Drugs available for the treatment of HAT have significant issues related to difficult administration regimes and limited efficacy across species and disease stages. Hence, there is considerable need to find new alternative and less toxic drugs. An approach to identify starting points for new drug candidates is high throughput screening (HTS of large compound library collections. We describe the application of an Alamar Blue based, 384-well HTS assay to screen a library of 87,296 compounds against the related trypanosome subspecies, Trypanosoma brucei brucei bloodstream form lister 427. Primary hits identified against T.b. brucei were retested and the IC(50 value compounds were estimated for T.b. brucei and a mammalian cell line HEK293, to determine a selectivity index for each compound. The screening campaign identified 205 compounds with greater than 10 times selectivity against T.b. brucei. Cluster analysis of these compounds, taking into account chemical and structural properties required for drug-like compounds, afforded a panel of eight compounds for further biological analysis. These compounds had IC(50 values ranging from 0.22 µM to 4 µM with associated selectivity indices ranging from 19 to greater than 345. Further testing against T.b. rhodesiense led to the selection of 6 compounds from 5 new chemical classes with activity against the causative species of HAT, which can be considered potential candidates for HAT early drug discovery. Structure activity relationship (SAR mining revealed components of those hit compound structures that may be important for biological activity. Four of these compounds have undergone further testing to 1 determine whether they are cidal or static in vitro at the minimum inhibitory concentration (MIC, and 2 estimate the time to kill.

  7. Importance of Donor Chondrocyte Viability for Osteochondral Allografts.

    Science.gov (United States)

    Cook, James L; Stannard, James P; Stoker, Aaron M; Bozynski, Chantelle C; Kuroki, Keiichi; Cook, Cristi R; Pfeiffer, Ferris M

    2016-05-01

    Osteochondral allograft (OCA) transplantation provides a biological treatment option for functional restoration of large articular cartilage defects in multiple joints. While successful outcomes after OCA transplantation have been linked to viable donor chondrocytes, the importance of donor cell viability has not been comprehensively validated. To use a canine model to determine the importance of donor chondrocyte viability at the time of implantation with respect to functional success of femoral condylar OCAs based on radiographic, gross, cell viability, histologic, biochemical, and biomechanical outcome measures. Controlled laboratory study. After approval was obtained from the institutional animal care and use committee, adult female dogs (N = 16) were implanted with 8-mm cylindrical OCAs from male dogs in the lateral and medial femoral condyles of 1 knee. OCAs were preserved for 28 or 60 days after procurement, and chondrocyte viability was quantified before implantation. Two different storage media, temperatures, and time points were used to obtain a spectrum of percentage chondrocyte viability at the time of implantation. A successful outcome was defined as an OCA that was associated with graft integration, maintenance of hyaline cartilage, lack of associated cartilage disorder, and lack of fibrillation, fissuring, or fibrous tissue infiltration of the allograft based on subjective radiographic, gross, and histologic assessments at 6 months after implantation. Chondrocyte viability ranged from 23% to 99% at the time of implantation. All successful grafts had >70% chondrocyte viability at the time of implantation, and no graft with chondrocyte viability <70% was associated with a successful outcome. Live-dead stained sections and histologic findings with respect to cell morphological features suggested that successful grafts were consistently composed of viable chondrocytes in lacunae, while grafts that were not successful were composed of nonviable

  8. Economic viability of anaerobic digestion

    Energy Technology Data Exchange (ETDEWEB)

    Wellinger, A. [INFOENERGIE, Ettenhausen (Switzerland)

    1996-01-01

    The industrial application of anaerobic digestion is a relatively new, yet proven waste treatment technology. Anaerobic digestion reduces and upgrades organic waste, and is a good way to control air pollution as it reduces methane and nitrous gas emissions. For environmental and energy considerations, anaerobic digestion is a nearly perfect waste treatment process. However, its economic viability is still in question. A number of parameters - type of waste (solid or liquid), digester system, facility size, product quality and end use, environmental requirements, cost of alternative treatments (including labor), and interest rates - define the investment and operating costs of an anaerobic digestion facility. Therefore, identical facilities that treat the same amount and type of waste may, depending on location, legislation, and end product characteristics, reveal radically different costs. A good approach for evaluating the economics of anaerobic digestion is to compare it to treatment techniques such as aeration or conventional sewage treatment (for industrial wastewater), or composting and incineration (for solid organic waste). For example, the cost (per ton of waste) of in-vessel composting with biofilters is somewhat higher than that of anaerobic digestion, but the investment costs 1 1/2 to 2 times more than either composting or anaerobic digestion. Two distinct advantages of anaerobic digestion are: (1) it requires less land than either composting or incinerating, which translates into lower costs and milder environmental and community impacts (especially in densely populated areas); and (2) it produces net energy, which can be used to operate the facility or sold to nearby industries.

  9. Viability of Event Management Business in Batangas City, Philippine: Basis for Business Operation Initiatives

    OpenAIRE

    Jeninah Christia D. Borbon

    2016-01-01

    The research study on Viability of Event Management Business in Batangas City: Basis for Business Operation Initiatives aimed to assess the viability of this type of business using Thompson’s (2005) Dimension of Business Viability as its tool in order to create business operation initiatives. It provided a good framework for defining success factors in entrepreneurial operation initiatives in a specific business type – event management. This study utilized event organizers based i...

  10. Linking population viability, habitat suitability, and landscape simulation models for conservation planning

    Science.gov (United States)

    Michael A. Larson; Frank R., III Thompson; Joshua J. Millspaugh; William D. Dijak; Stephen R. Shifley

    2004-01-01

    Methods for habitat modeling based on landscape simulations and population viability modeling based on habitat quality are well developed, but no published study of which we are aware has effectively joined them in a single, comprehensive analysis. We demonstrate the application of a population viability model for ovenbirds (Seiurus aurocapillus)...

  11. Pollen viability and membrane lipid composition

    NARCIS (Netherlands)

    Bilsen, van D.G.J.L.

    1993-01-01

    In this thesis membrane lipid composition is studied in relation to pollen viability during storage. Chapter 1 reviews pollen viability, membranes in the dry state and membrane changes associated with cellular aging. This chapter is followed by a study of age-related changes in phospholipid

  12. Third-Generation Ah Receptor–Responsive Luciferase Reporter Plasmids: Amplification of Dioxin-Responsive Elements Dramatically Increases CALUX Bioassay Sensitivity and Responsiveness

    Science.gov (United States)

    He, Guochun; Tsutsumi, Tomoaki; Zhao, Bin; Baston, David S.; Zhao, Jing; Heath-Pagliuso, Sharon; Denison, Michael S.

    2011-01-01

    2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related dioxin-like chemicals are widespread and persistent environmental contaminants that produce diverse toxic and biological effects through their ability to bind to and activate the Ah receptor (AhR) and AhR-dependent gene expression. The chemically activated luciferase expression (CALUX) system is an AhR-responsive recombinant luciferase reporter gene–based cell bioassay that has been used in combination with chemical extraction and cleanup methods for the relatively rapid and inexpensive detection and relative quantitation of dioxin and dioxin-like chemicals in a wide variety of sample matrices. Although the CALUX bioassay has been validated and used extensively for screening purposes, it has some limitations when screening samples with very low levels of dioxin-like chemicals or when there is only a small amount of sample matrix for analysis. Here, we describe the development of third-generation (G3) CALUX plasmids with increased numbers of dioxin-responsive elements, and stable transfection of these new plasmids into mouse hepatoma (Hepa1c1c7) cells has produced novel amplified G3 CALUX cell bioassays that respond to TCDD with a dramatically increased magnitude of luciferase induction and significantly lower minimal detection limit than existing CALUX-type cell lines. The new G3 CALUX cell lines provide a highly responsive and sensitive bioassay system for the detection and relative quantitation of very low levels of dioxin-like chemicals in sample extracts. PMID:21775728

  13. Proof of Economic Viability of Blended Learning Business Models

    Science.gov (United States)

    Druhmann, Carsten; Hohenberg, Gregor

    2014-01-01

    The discussion on economically sustainable business models with respect to information technology is lacking in many aspects of proven approaches. In the following contribution the economic viability is valued based on a procedural model for design and evaluation of e-learning business models in the form of a case study. As a case study object a…

  14. Steady-state fluorescence and phosphorescence spectroscopic studies of bacterial luciferase tryptophan mutants.

    Science.gov (United States)

    Li, Z; Meighen, E A

    1994-09-01

    Bacterial luciferase, which catalyzes the bioluminescence reaction in luminous bacteria, consists of two nonidentical polypeptides, α and β. Eight mutants of luciferase with each of the tryptophans replaced by tyrosine were generated by site-directed mutagenesis and purified to homogeneity. The steady-state tryptophan fluorescence and low-temperature phosphorescence spectroscopic properties of these mutants were characterized. In some instances, mutation of only a single tryptophan residue resulted in large spectral changes. The tryptophan residues conserved in both the α and the β subunits exhibited distinct fluorescence emission properties, suggesting that these tryptophans have different local enviroments. The low-temperature phosphorescence data suggest that the tryptophans conserved in bot the α and the β subunits are not located at the subunit interface and/or involved in subunit interactions. The differences in the spectral properties of the mutants have provided useful information on the local environment of the individual tryptophan residues as well as on the quaternary structure of the protein.

  15. Luciferase NanoLuc as a reporter for gene expression and protein levels in Saccharomyces cerevisiae.

    Science.gov (United States)

    Masser, Anna E; Kandasamy, Ganapathi; Kaimal, Jayasankar Mohanakrishnan; Andréasson, Claes

    2016-05-01

    Reporter proteins are essential tools in the study of biological processes and are employed to monitor changes in gene expression and protein levels. Luciferases are reporter proteins that enable rapid and highly sensitive detection with an outstanding dynamic range. Here we evaluated the usefulness of the 19 kDa luciferase NanoLuc (Nluc), derived from the deep sea shrimp Oplophorus gracilirostris, as a reporter protein in yeast. Cassettes with codon-optimized genes expressing yeast Nluc (yNluc) or its destabilized derivative yNlucPEST have been assembled in the context of the dominant drug resistance marker kanMX. The reporter proteins do not impair the growth of yeast cells and exhibit half-lives of 40 and 5 min, respectively. The commercial substrate Nano-Glo® is compatible with detection of yNluc bioluminescence in yeast using standard commercial substrate. © 2016 The Authors. Yeast published by John Wiley & Sons Ltd. © 2016 The Authors. Yeast published by John Wiley & Sons Ltd.

  16. Doctors' perspectives on the viability of rural practice.

    Science.gov (United States)

    Jones, J A; Humphreys, J S; Adena, M A

    2004-01-01

    Private practitioners play a vital role in meeting the health needs of rural communities. However, the prospect of operating a private practice business in rural Australia seems to be increasingly unattractive, because many communities are forced to recruit salaried or overseas-trained doctors. This study focuses on rural practices as businesses whose viability influences their attractiveness for the recruitment and retention of practitioners. The specific objectives are to ascertain which factors contribute to or threaten practice viability in rural areas, and whether they vary according to the degree of rurality or geographical remoteness. This study is based on data collected from a national study into the viability of rural general practice undertaken jointly by the Rural Doctors Association of Australia and Monash University School of Rural Health Bendigo. The Rural Remote and Metropolitan Area (RRMA) classification was used as the indicator of rurality. The study surveyed all general practitioners practising in rural or remote regions of Australia (RRMAs 3 to 7). Only practitioners with some financial interest in the practice were selected for this analysis. Free-text responses to the two questions 'What are the key factors contributing to the viability of your practice?' and 'What factors would put the viability of your practice at risk?' were analysed using qualitative content analysis. Factors were derived iteratively through higher-level aggregation of responses. Chi-square tests were used to make comparisons across the RRMA categories. The national survey achieved a response rate of 35% of the entire population of GPs practising in RRMA 3 to 7 regions. Of these, 1050 respondents were relevant to this analysis. Seven major factors were identified by practitioners as the main contributors to practice viability. 'Practice characteristics' was nominated by 59% of respondents, followed by 'Income' (31%), 'Personal circumstances', 'Workforce' and 'Community

  17. A novel luciferase knock-in reporter system for studying transcriptional regulation of the human Sox2 gene.

    Science.gov (United States)

    Xiao, Dan; Zhang, Weifeng; Li, Yan; Liu, Kuan; Zhao, Junli; Sun, Xiaohong; Shan, Linlin; Mao, Qinwen; Xia, Haibin

    2016-02-10

    Sox2 is an important transcriptional factor that has multiple functions in stem cell maintenance and tumorigenesis. To investigate the transcriptional regulation of the Sox2 gene, a luciferase knock-in reporter system was established in HEK293 cells by placing the luciferase gene in the genome under the control of the Sox2 gene promoter using a transcription activator-like effector nuclease (TALEN)-mediated genome editing technique. PCR and Southern blot results confirmed the site-specific integration of a single copy of the exogenous luciferase gene into the genome. To prove the reliability and sensitivity of this novel luciferase knock-in system, a CRISPR/Cas transcription activation system for the Sox2 gene was constructed and applied to the knock-in system. The results indicated that luciferase activity was directly correlated with the activity of the Sox2 endogenous promoter. This novel system will be a useful tool to study the transcriptional regulation of Sox2, and has great potential in medical and industrial applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Population-specific life histories contribute to metapopulation viability

    Science.gov (United States)

    Halsey, Samniqueka J.; Bell, Timothy J.; McEachern, A. Kathryn; Pavlovic, Noel B.

    2016-01-01

    metapopulation viability. In understanding the underlying causes of the projected extinction probabilities of each population and identifying broad-scale contributions of different populations to the metapopulation, the process of pinpointing target populations is simplified. More detailed analyses can then be applied to the target populations to increase population viability and consequently metapopulation viability. Based on our research, we suggest that the best approach to improve the overall metapopulation viability is to manage the contributions to population growth for each population separately.

  19. Specific detection of peste des petits ruminants virus antibodies in sheep and goat sera by the luciferase

    International Nuclear Information System (INIS)

    Berguido, F.J.; Bodjo, S.C.; Loitsch, A.; Diallo, A.

    2016-01-01

    Full text: Peste des petits ruminants (PPR) is a contagious and often fatal transboundary animal disease affecting mostly sheep, goats and wild small ruminants. This disease is endemic in most of Africa, the Middle, Near East, and large parts of Asia. The casual agent is peste des petits ruminants virus (PPRV), which belongs to the genus Morbilivirus in the family Paramyxoviridae. This genus also includes measles virus (MV), canine distemper virus (CDV) and rinderpest virus (RPV). All are closely related viruses with serological cross reactivity. In this study, we have developed a Luciferase Immunoprecipitation System (LIPS) for the rapid detection of antibodies against PPRV in serum samples and for specific differentiation from antibodies against RPV. PPR and rinderpest (RP) serum samples were assayed by PPR-LIPS and two commercially available PPR cELISA tests. The PPR-LIPS showed high sensitivity and specificity for the samples tested and showed no cross reactivity with RPV unlike the commercial PPR cELISA tests which did not cross react with RPV. Based on the results shown in this study, PPR-LIPS is presented as a good candidate for the specific serosurveillance of PPR. (author)

  20. Development and Validation of a Novel Dual Luciferase Reporter Gene Assay to Quantify Ebola Virus VP24 Inhibition of IFN Signaling

    Directory of Open Access Journals (Sweden)

    Elisa Fanunza

    2018-02-01

    Full Text Available The interferon (IFN system is the first line of defense against viral infections. Evasion of IFN signaling by Ebola viral protein 24 (VP24 is a critical event in the pathogenesis of the infection and, hence, VP24 is a potential target for drug development. Since no drugs target VP24, the identification of molecules able to inhibit VP24, restoring and possibly enhancing the IFN response, is a goal of concern. Accordingly, we developed a dual signal firefly and Renilla luciferase cell-based drug screening assay able to quantify IFN-mediated induction of Interferon Stimulated Genes (ISGs and its inhibition by VP24. Human Embryonic Kidney 293T (HEK293T cells were transiently transfected with a luciferase reporter gene construct driven by the promoter of ISGs, Interferon-Stimulated Response Element (ISRE. Stimulation of cells with IFN-α activated the IFN cascade leading to the expression of ISRE. Cotransfection of cells with a plasmid expressing VP24 cloned from a virus isolated during the last 2014 outbreak led to the inhibition of ISRE transcription, quantified by a luminescent signal. To adapt this system to test a large number of compounds, we performed it in 96-well plates; optimized the assay analyzing different parameters; and validated the system by calculating the Z′- and Z-factor, which showed values of 0.62 and 0.53 for IFN-α stimulation assay and VP24 inhibition assay, respectively, indicative of robust assay performance.

  1. Super RLuc8: A novel engineered Renilla luciferase with a red-shifted spectrum and stable light emission.

    Science.gov (United States)

    Rahnama, Somaieh; Saffar, Behnaz; Kahrani, Zahra Fanaei; Nazari, Mahboobeh; Emamzadeh, Rahman

    2017-01-01

    Renilla luciferase is a bioluminescent enzyme which is broadly used as a reporter protein in molecular biosensors. In this study, a novel luciferase with desired light emission wavelength and thermostability is reported. The results indicated that the new luciferase, namely super RLuc8, had a red-shifted spectrum and showed stable light emission. Super RLuc8 showed a 10-fold (p-value=0.0084) increase in the thermostability at 37°C after 20min incubation, in comparison to the native enzyme. The optimum temperature of the mutant increased from 30 to 37°C. Molecular dynamics simulation analysis indicated that the increased thermostability was most probably caused by a better structural compactness and more local rigidity in the regions out of the emitter site. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Economic Viability and Marketing Strategies of Periwinkle ...

    African Journals Online (AJOL)

    Economic Viability and Marketing Strategies of Periwinkle Tympanotonus Fuscatus in Rivers State, Nigeria. ... The results indicated that marketing strategies are enroute, through harvesters (collectors), ... EMAIL FULL TEXT EMAIL FULL TEXT

  3. Supply Chain Viability for the North American Microwave Power Tube Industry

    National Research Council Canada - National Science Library

    Philippi, Therese M

    2002-01-01

    The DoD Defense Production (DPA) Act Title III program sponsored this project in response to a critical supply base issue with the objective of strengthening the supplier base to ensure it's future viability...

  4. Storage Viability and Optimization Web Service

    Energy Technology Data Exchange (ETDEWEB)

    Stadler, Michael; Marnay, Christ; Lai, Judy; Siddiqui, Afzal; Limpaitoon, Tanachai; Phan, Trucy; Megel, Olivier; Chang, Jessica; DeForest, Nicholas

    2010-10-11

    Non-residential sectors offer many promising applications for electrical storage (batteries) and photovoltaics (PVs). However, choosing and operating storage under complex tariff structures poses a daunting technical and economic problem that may discourage potential customers and result in lost carbon and economic savings. Equipment vendors are unlikely to provide adequate environmental analysis or unbiased economic results to potential clients, and are even less likely to completely describe the robustness of choices in the face of changing fuel prices and tariffs. Given these considerations, researchers at Lawrence Berkeley National Laboratory (LBNL) have designed the Storage Viability and Optimization Web Service (SVOW): a tool that helps building owners, operators and managers to decide if storage technologies and PVs merit deeper analysis. SVOW is an open access, web-based energy storage and PV analysis calculator, accessible by secure remote login. Upon first login, the user sees an overview of the parameters: load profile, tariff, technologies, and solar radiation location. Each parameter has a pull-down list of possible predefined inputs and users may upload their own as necessary. Since the non-residential sectors encompass a broad range of facilities with fundamentally different characteristics, the tool starts by asking the users to select a load profile from a limited cohort group of example facilities. The example facilities are categorized according to their North American Industry Classification System (NAICS) code. After the load profile selection, users select a predefined tariff or use the widget to create their own. The technologies and solar radiation menus operate in a similar fashion. After these four parameters have been inputted, the users have to select an optimization setting as well as an optimization objective. The analytic engine of SVOW is LBNL?s Distributed Energy Resources Customer Adoption Model (DER-CAM), which is a mixed

  5. Evaluation of Gaussia luciferase and foot-and-mouth disease virus 2A translational interrupter chimeras as polycistronic reporters for transgene expression.

    Science.gov (United States)

    Puckette, Michael; Burrage, Thomas; Neilan, John G; Rasmussen, Max

    2017-06-12

    The Gaussia princeps luciferase is used as a stand-alone reporter of transgene expression for in vitro and in vivo expression systems due to the rapid and easy monitoring of luciferase activity. We sought to simultaneously quantitate production of other recombinant proteins by transcriptionally linking the Gaussia princeps luciferase gene to other genes of interest through the foot-and-mouth disease virus 2A translational interrupter sequence. We produced six plasmids, each encoding a single open reading frame, with the foot-and-mouth disease virus 2A sequence placed either N-terminal or C-terminal to the Gaussia princeps luciferase gene. Two plasmids included novel Gaussia princeps luciferase variants with the position 1 methionine deleted. Placing a foot-and-mouth disease virus 2A translational interrupter sequence on either the N- or C-terminus of the Gaussia princeps luciferase gene did not prevent the secretion or luminescence of resulting chimeric luciferase proteins. We also measured the ability of another polycistronic plasmid vector with a 2A-luciferase sequence placed downstream of the foot-and-mouth disease virus P1 and 3C protease genes to produce of foot-and-mouth disease virus-like particles and luciferase activity from transfected cells. Incorporation of the 2A-luciferase sequence into a transgene encoding foot-and-mouth disease virus structural proteins retained luciferase activity and the ability to form virus-like particles. We demonstrated a mechanism for the near real-time, sequential, non-destructive quantitative monitoring of transcriptionally-linked recombinant proteins and a valuable method for monitoring transgene expression in recombinant vaccine constructs.

  6. Viability of lactobacillus acidophilus in various vaginal tablet formulations

    Directory of Open Access Journals (Sweden)

    Fazeli M.R.

    2006-07-01

    Full Text Available The lactobacilli which are present in vaginal fluids play an important role in prevention of vaginosis and there are considerable interests in formulation of these friendly bacteria into suitable pharmaceutical dosage forms. Formulating these microorganisms for vaginal application is a critical issue as the products should retain viability of lactobacilli during formulation and also storage. The aim of this study was to examine the viability and release of Lactobacillus acidophilus from slow-release vaginal tablets prepared by using six different retarding polymers and from two effervescent tablets prepared by using citric or adipic acid. The Carbomer–based formulations showed high initial viablility compared to those based on HPMC-LV, HPMC-HV, Polycarbophil and SCMC polymers which showed one log decrease in viable cells. All retarding polymers in slow release formulations presented a strong bacterial release at about 2 h except Carbomer polymers which showed to be poor bacterial releasers. Although effervescent formulations produced a quick bacterial release in comparison with polymer based slow-release tablets, they were less stable in cold storage. Due to the strong chelating characteristic of citric acid, the viability was quickly lost for aqueous medium of citric acid in comparison with adipic acid based effervescent tablets.

  7. Assessment of myocardial viability by MR imaging

    International Nuclear Information System (INIS)

    Sandstede, Joern J.W.

    2003-01-01

    Diagnosis of myocardial viability after infarction focuses on the prediction of functional improvement of dysfunctional myocardium after revascularization therapy. Magnetic resonance imaging provides different approaches for the detection of myocardial viability. Measurement of end-diastolic wall thickness is easy to perform and has a high sensitivity, but a low specificity, and can only be used 4 months after myocardial infarction due to infarct healing processes. Low-dose dobutamine stress has a good sensitivity with a high specificity for the prediction of wall motion improvement, but this is only true for patients with a singular dysfunctional area and only slightly depressed cardiac function. Late enhancement allows for direct visualization of necrotic or scarred tissue. By measuring the transmural extent of late enhancement, the probability of mechanical improvement can precisely be given. Imaging of microvascular obstruction by first-pass perfusion or late enhancement gives additional information on viability and patient prognosis. Metabolic imaging techniques, such as 31 P-MR spectroscopy and 23 Na-MR imaging, provide further insights into the mechanisms of myocardial infarction and viability. In conclusion, cardiac MRI offers several clinically usable approaches for the assessment of myocardial viability and will probably become the method of choice in the near future. (orig.)

  8. LIVE/DEAD YEAST VIABILITY STAINING AS A TOOL FOR IMPROVING ARTISANAL PILSNER BEER PRODUCTION

    Directory of Open Access Journals (Sweden)

    Benedetta Bottari

    2014-10-01

    Full Text Available The production of an artisanal beer, made by brewers using traditional practices on a small scale, is founded on the empirical adjustment of parameters, including yeasts handling and serial repitching. The aim of this study was to monitor yeast viability during different stages of artisanal beer productions through the Live/Dead Yeast viability staining and to correlate it with fermentation dynamics in order to increase process standardization and to maintain the quality of final products. Yeast viability and fermentation activities were evaluated during seven fermentation cycles of an artisanal pilsner beer. Yeast inoculated with higher viability performed generally better in fermentation, resulting in faster sugar consumption, faster ethanol production and stability. Handling yeast and serial repitching based on Live/Dead viability measurements, could be the key way to ensure reliable manufacture of high quality beer and to improve process standardization particularly for microbreweries, where variability of production can be a challenging point.

  9. Population viability analysis for endangered Roanoke logperch

    Science.gov (United States)

    Roberts, James H.; Angermeier, Paul; Anderson, Gregory B.

    2016-01-01

    A common strategy for recovering endangered species is ensuring that populations exceed the minimum viable population size (MVP), a demographic benchmark that theoretically ensures low long-term extinction risk. One method of establishing MVP is population viability analysis, a modeling technique that simulates population trajectories and forecasts extinction risk based on a series of biological, environmental, and management assumptions. Such models also help identify key uncertainties that have a large influence on extinction risk. We used stochastic count-based simulation models to explore extinction risk, MVP, and the possible benefits of alternative management strategies in populations of Roanoke logperch Percina rex, an endangered stream fish. Estimates of extinction risk were sensitive to the assumed population growth rate and model type, carrying capacity, and catastrophe regime (frequency and severity of anthropogenic fish kills), whereas demographic augmentation did little to reduce extinction risk. Under density-dependent growth, the estimated MVP for Roanoke logperch ranged from 200 to 4200 individuals, depending on the assumed severity of catastrophes. Thus, depending on the MVP threshold, anywhere from two to all five of the logperch populations we assessed were projected to be viable. Despite this uncertainty, these results help identify populations with the greatest relative extinction risk, as well as management strategies that might reduce this risk the most, such as increasing carrying capacity and reducing fish kills. Better estimates of population growth parameters and catastrophe regimes would facilitate the refinement of MVP and extinction-risk estimates, and they should be a high priority for future research on Roanoke logperch and other imperiled stream-fish species.

  10. Comparison of tissue viability imaging and colorimetry: skin blanching.

    Science.gov (United States)

    Zhai, Hongbo; Chan, Heidi P; Farahmand, Sara; Nilsson, Gert E; Maibach, Howard I

    2009-02-01

    Operator-independent assessment of skin blanching is important in the development and evaluation of topically applied steroids. Spectroscopic instruments based on hand-held probes, however, include elements of operator dependence such as difference in applied pressure and probe misalignment, while laser Doppler-based methods are better suited for demonstration of skin vasodilatation than for vasoconstriction. To demonstrate the potential of the emerging technology of Tissue Viability Imaging (TiVi) in the objective and operator-independent assessment of skin blanching. The WheelsBridge TiVi600 Tissue Viability Imager was used for quantification of human skin blanching with the Minolta chromameter CR 200 as an independent colorimeter reference method. Desoximetasone gel 0.05% was applied topically on the volar side of the forearm under occlusion for 6 h in four healthy adults. In a separate study, the induction of blanching in the occlusion phase was mapped using a transparent occlusion cover. The relative uncertainty in the blanching estimate produced by the Tissue Viability Imager was about 5% and similar to that of the chromameter operated by a single user and taking the a(*) parameter as a measure of blanching. Estimation of skin blanching could also be performed in the presence of a transient paradoxical erythema, using the integrated TiVi software. The successive induction of skin blanching during the occlusion phase could readily be mapped by the Tissue Viability Imager. TiVi seems to be suitable for operator-independent and remote mapping of human skin blanching, eliminating the main disadvantages of methods based on hand-held probes.

  11. Economic viability of Cernavoda project

    International Nuclear Information System (INIS)

    Condu, M.; Popescu, D.; Chirica, T.; Glodeanu, F.

    1998-01-01

    Cernavoda project started in 1978, when the CANDU technology procurement contract and other contracts for engineering, technical assistance and procurement were signed with AECL. The works on site started in 1980. In February 1981, a contract was signed with GE - USA and Ansaldo - Italy for balance of plant. After 1989 the concept for project implementation was essentially changed. The completion of Cernavoda NPP Unit-1, established as the first priority by Romanian authorities was implemented by AECL-Ansaldo Consortium (AAC). The objective of the contract signed with AAC consisted in overtaking the management of Cernavoda Unit-1 project, in order to complete, commission and operate the plant for 18 months. The plant was successfully completed, commissioned and commercially operated since December 2, 1996. As concerns Unit-2, only preservation and some remedial works were performed since 1991. In order to support the decision to complete Unit-2, a complete process was initiated: - identification of the activities and associated costs required for plan completion; - plant integration in the utility's least cost development study; - Unit-2 cash flow analysis. A joint RENEL-AECL-ANSALDO team performed a detailed analysis using Cernavoda Unit-1 as reference project and taking advantage of the experience gained during construction, commissioning and operation of the first unit. The least cost power generation development study finalized by SEP and EDF, based on a PHARE funding, ranked Cernavoda Unit-2 on the second place, after two small hydroelectric plants of 53 and 8 Mw e. A cash flow model developed by a RENEL-ANSALDO joint team shows that Cernavoda Unit-2 is an economically feasible project. The financing scenario provides that all resources will be from loans. Other arguments for completion of Cernavoda Unit-2 are: - good performances in operation of Unit-1; - the need to provide jobs for the specialists in the nuclear field; - opportunity for electricity export

  12. Traffic networks as information systems a viability approach

    CERN Document Server

    Aubin, Jean-Pierre

    2017-01-01

    This authored monograph covers a viability to approach to traffic management by advising to vehicles circulated on the network the velocity they should follow for satisfying global traffic conditions;. It presents an investigation of three structural innovations: The objective is to broadcast at each instant and at each position the advised celerity to vehicles, which could be read by auxiliary speedometers or used by cruise control devices. Namely, 1. Construct regulation feedback providing at each time and position advised velocities (celerities) for minimizing congestion or other requirements. 2. Taking into account traffic constraints of different type, the first one being to remain on the roads, to stop at junctions, etc. 3. Use information provided by the probe vehicles equipped with GPS to the traffic regulator; 4. Use other global traffic measures of vehicles provided by different types of sensors; These results are based on convex analysis, intertemporal optimization and viability theory as mathemati...

  13. Cost viability of 3D printed house in UK

    Science.gov (United States)

    Tobi, A. L. Mohd; Omar, S. A.; Yehia, Z.; Al-Ojaili, S.; Hashim, A.; Orhan, O.

    2018-03-01

    UK has been facing housing crisis due to the rising price of the property on sale. This paper will look into the viability of 3D printing technology as an alternative way for house construction on UK. The analysis will be carried out based on the data until the year of 2014 due to limited resources availability. Details cost breakdown on average size house construction cost in UK were analysed and relate to the cost viability of 3D printing technology in reducing the house price in UK. It is found that the 3D printing generates saving of up to around 35% out of total house price in UK. This cost saving comes from the 3D printed construction of walls and foundations for material and labour cost.

  14. Population Viability of Avian Endangered Species: the PVAvES Program

    National Research Council Canada - National Science Library

    Melton, Robert

    2001-01-01

    .... The program is designed to assess the viability of endangered bird species populations on U.S. Army lands. It also facilitates the comparison of alternative ecological scenarios based on different assumptions about the effects of natural or human...

  15. Assessment of tissue viability by polarization spectroscopy

    Science.gov (United States)

    Nilsson, G.; Anderson, C.; Henricson, J.; Leahy, M.; O'Doherty, J.; Sjöberg, F.

    2008-09-01

    A new and versatile method for tissue viability imaging based on polarization spectroscopy of blood in superficial tissue structures such as the skin is presented in this paper. Linearly polarized light in the visible wavelength region is partly reflected directly by the skin surface and partly diffusely backscattered from the dermal tissue matrix. Most of the directly reflected light preserves its polarization state while the light returning from the deeper tissue layers is depolarized. By the use of a polarization filter positioned in front of a sensitive CCD-array, the light directly reflected from the tissue surface is blocked, while the depolarized light returning from the deeper tissue layers reaches the detector array. By separating the colour planes of the detected image, spectroscopic information about the amount of red blood cells (RBCs) in the microvascular network of the tissue under investigation can be derived. A theory that utilizes the differences in light absorption of RBCs and bloodless tissue in the red and green wavelength region forms the basis of an algorithm for displaying a colour coded map of the RBC distribution in a tissue. Using a fluid model, a linear relationship (cc. = 0.99) between RBC concentration and the output signal was demonstrated within the physiological range 0-4%. In-vivo evaluation using transepidermal application of acetylcholine by the way of iontophoresis displayed the heterogeneity pattern of the vasodilatation produced by the vasoactive agent. Applications of this novel technology are likely to be found in drug and skin care product development as well as in the assessment of skin irritation and tissue repair processes and even ultimately in a clinic case situation.

  16. Molecular assays for determining Mycobacterium leprae viability in tissues of experimentally infected mice.

    Science.gov (United States)

    Davis, Grace L; Ray, Nashone A; Lahiri, Ramanuj; Gillis, Thomas P; Krahenbuhl, James L; Williams, Diana L; Adams, Linda B

    2013-01-01

    The inability of Mycobacterium leprae to grow on axenic media has necessitated specialized techniques in order to determine viability of this organism. The purpose of this study was to develop a simple and sensitive molecular assay for determining M. leprae viability directly from infected tissues. Two M. leprae-specific quantitative reverse transcription PCR (qRT-PCR) assays based on the expression levels of esxA, encoding the ESAT-6 protein, and hsp18, encoding the heat shock 18 kDa protein, were developed and tested using infected footpad (FP) tissues of both immunocompetent and immunocompromised (athymic nu/nu) mice. In addition, the ability of these assays to detect the effects of anti-leprosy drug treatment on M. leprae viability was determined using rifampin and rifapentine, each at 10 mg/kg for 1, 5, or 20 daily doses, in the athymic nu/nu FP model. Molecular enumeration (RLEP PCR) and viability determinations (qRT-PCR) were performed via Taqman methodology on DNA and RNA, respectively, purified from ethanol-fixed FP tissue and compared with conventional enumeration (microscopic counting of acid fast bacilli) and viability assays (radiorespirometry, viability staining) which utilized bacilli freshly harvested from the contralateral FP. Both molecular and conventional assays demonstrated growth and high viability of M. leprae in nu/nu FPs over a 4 month infection period. In contrast, viability was markedly decreased by 8 weeks in immunocompetent mice. Rifapentine significantly reduced bacterial viability after 5 treatments, whereas rifampin required up to 20 treatments for the same efficacy. Neither drug was effective after a single treatment. In addition, host gene expression was monitored with the same RNA preparations. hsp18 and esxA qRT-PCR are sensitive molecular indicators, reliably detecting viability of M. leprae in tissues without the need for bacterial isolation or immediate processing, making these assays applicable for in vivo drug screening and

  17. Viability of mesenchymal stem cells during electrospinning

    Directory of Open Access Journals (Sweden)

    G. Zanatta

    2012-02-01

    Full Text Available Tissue engineering is a technique by which a live tissue can be re-constructed and one of its main goals is to associate cells with biomaterials. Electrospinning is a technique that facilitates the production of nanofibers and is commonly used to develop fibrous scaffolds to be used in tissue engineering. In the present study, a different approach for cell incorporation into fibrous scaffolds was tested. Mesenchymal stem cells were extracted from the wall of the umbilical cord and mononuclear cells from umbilical cord blood. Cells were re-suspended in a 10% polyvinyl alcohol solution and subjected to electrospinning for 30 min under a voltage of 21 kV. Cell viability was assessed before and after the procedure by exclusion of dead cells using trypan blue staining. Fiber diameter was observed by scanning electron microscopy and the presence of cells within the scaffolds was analyzed by confocal laser scanning microscopy. After electrospinning, the viability of mesenchymal stem cells was reduced from 88 to 19.6% and the viability of mononuclear cells from 99 to 8.38%. The loss of viability was possibly due to the high viscosity of the polymer solution, which reduced the access to nutrients associated with electric and mechanical stress during electrospinning. These results suggest that the incorporation of cells during fiber formation by electrospinning is a viable process that needs more investigation in order to find ways to protect cells from damage.

  18. Construction and characterization of a recombinant yellow fever virus stably expressing Gaussia luciferase

    Directory of Open Access Journals (Sweden)

    TELISSA C. KASSAR

    Full Text Available ABSTRACT Yellow fever is an arthropod-borne viral disease that still poses high public health concerns, despite the availability of an effective vaccine. The development of recombinant viruses is of utmost importance for several types of studies, such as those aimed to dissect virus-host interactions and to search for novel antiviral strategies. Moreover, recombinant viruses expressing reporter genes may greatly facilitate these studies. Here, we report the construction of a recombinant yellow fever virus (YFV expressing Gaussia luciferase (GLuc (YFV-GLuc. We show, through RT-PCR, sequencing and measurement of GLuc activity, that stability of the heterologous gene was maintained after six passages. Furthermore, a direct association between GLuc expression and viral replication was observed (r2=0.9967, indicating that measurement of GLuc activity may be used to assess viral replication in different applications. In addition, we evaluated the use of the recombinant virus in an antiviral assay with recombinant human alfa-2b interferon. A 60% inhibition of GLuc expression was observed in cells infected with YFV-GLuc and incubated with IFN alfa-2b. Previously tested on YFV inhibition by plaque assays indicated a similar fold-decrease in viral replication. These results are valuable as they show the stability of YFV-GLuc and one of several possible applications of this construct.

  19. Autonomous bioluminescent expression of the bacterial luciferase gene cassette (lux in a mammalian cell line.

    Directory of Open Access Journals (Sweden)

    Dan M Close

    Full Text Available The bacterial luciferase (lux gene cassette consists of five genes (luxCDABE whose protein products synergistically generate bioluminescent light signals exclusive of supplementary substrate additions or exogenous manipulations. Historically expressible only in prokaryotes, the lux operon was re-synthesized through a process of multi-bicistronic, codon-optimization to demonstrate for the first time self-directed bioluminescence emission in a mammalian HEK293 cell line in vitro and in vivo.Autonomous in vitro light production was shown to be 12-fold greater than the observable background associated with untransfected control cells. The availability of reduced riboflavin phosphate (FMNH(2 was identified as the limiting bioluminescence substrate in the mammalian cell environment even after the addition of a constitutively expressed flavin reductase gene (frp from Vibrio harveyi. FMNH(2 supplementation led to a 151-fold increase in bioluminescence in cells expressing mammalian codon-optimized luxCDE and frp genes. When injected subcutaneously into nude mice, in vivo optical imaging permitted near instantaneous light detection that persisted independently for the 60 min length of the assay with negligible background.The speed, longevity, and self-sufficiency of lux expression in the mammalian cellular environment provides a viable and powerful alternative for real-time target visualization not currently offered by existing bioluminescent and fluorescent imaging technologies.

  20. Probing Bioluminescence Resonance Energy Transfer in Quantum Rod-Luciferase Nanoconjugates.

    Science.gov (United States)

    Alam, Rabeka; Karam, Liliana M; Doane, Tennyson L; Coopersmith, Kaitlin; Fontaine, Danielle M; Branchini, Bruce R; Maye, Mathew M

    2016-02-23

    We describe the necessary design criteria to create highly efficient energy transfer conjugates containing luciferase enzymes derived from Photinus pyralis (Ppy) and semiconductor quantum rods (QRs) with rod-in-rod (r/r) microstructure. By fine-tuning the synthetic conditions, CdSe/CdS r/r-QRs were prepared with two different emission colors and three different aspect ratios (l/w) each. These were hybridized with blue, green, and red emitting Ppy, leading to a number of new BRET nanoconjugates. Measurements of the emission BRET ratio (BR) indicate that the resulting energy transfer is highly dependent on QR energy accepting properties, which include absorption, quantum yield, and optical anisotropy, as well as its morphological and topological properties, such as aspect ratio and defect concentration. The highest BR was found using r/r-QRs with lower l/w that were conjugated with red Ppy, which may be activating one of the anisotropic CdSe core energy levels. The role QR surface defects play on Ppy binding, and energy transfer was studied by growth of gold nanoparticles at the defects, which indicated that each QR set has different sites. The Ppy binding at those sites is suggested by the observed BRET red-shift as a function of Ppy-to-QR loading (L), where the lowest L results in highest efficiency and furthest shift.

  1. Detecting viability transitions of umbilical cord mesenchymal stem cells by Raman micro-spectroscopy

    International Nuclear Information System (INIS)

    Bai, H; Chen, P; Fang, H; Lin, L; Tang, G Q; Mu, G G; Gong, W; Liu, Z P; Wu, H; Zhao, H; Han, Z C

    2011-01-01

    Recent research suggests that human umbilical cord derived mesenchymal stem cells (hUC-MSCs) can be promising candidates for cell-based therapy. Since large population and high viability are generally required, detecting viability transitions of these cells is crucial for their population expansion and quality control. Here, as a non-invasive method, Raman micro-spectroscopy is applied to examine hUC-MSCs with different viability. Using peak fitting and statistic t-test, the Raman peaks with obvious differences between the cells with high viability (> 90%) and low viability ( -1 , symmetric stretching of C–C in lipids at 877 cm -1 and CH deformation in proteins at 1342 cm -1 show the most significant changes (p < 0.001). When the cell viability decreases, the intensities of the former two peaks are both about doubled while that of the latter peak reduces by about 30%. Based on these results, we propose that the viability of hUC-MSCs can be characterized by these three peaks. And their intensity changes can be understood from the model of excessive reactive oxygen species interacting with the bio-macromolecules

  2. Sphingosine kinase activity is not required for tumor cell viability.

    Directory of Open Access Journals (Sweden)

    Karen Rex

    Full Text Available Sphingosine kinases (SPHKs are enzymes that phosphorylate the lipid sphingosine, leading to the formation of sphingosine-1-phosphate (S1P. In addition to the well established role of extracellular S1P as a mitogen and potent chemoattractant, SPHK activity has been postulated to be an important intracellular regulator of apoptosis. According to the proposed rheostat theory, SPHK activity shifts the intracellular balance from the pro-apoptotic sphingolipids ceramide and sphingosine to the mitogenic S1P, thereby determining the susceptibility of a cell to apoptotic stress. Despite numerous publications with supporting evidence, a clear experimental confirmation of the impact of this mechanism on tumor cell viability in vitro and in vivo has been hampered by the lack of suitable tool reagents. Utilizing a structure based design approach, we developed potent and specific SPHK1/2 inhibitors. These compounds completely inhibited intracellular S1P production in human cells and attenuated vascular permeability in mice, but did not lead to reduced tumor cell growth in vitro or in vivo. In addition, siRNA experiments targeting either SPHK1 or SPHK2 in a large panel of cell lines failed to demonstrate any statistically significant effects on cell viability. These results show that the SPHK rheostat does not play a major role in tumor cell viability, and that SPHKs might not be attractive targets for pharmacological intervention in the area of oncology.

  3. Effect of Isolation Techniques on Viability of Bovine Blood Neutrophils

    Directory of Open Access Journals (Sweden)

    P. Sláma

    2006-01-01

    Full Text Available The effect of selected isolation methods on the viability of neutrophil granulocytes (neutrophils from the blood of healthy Holstein x Bohemian Red Pied crossbred heifers was evaluated. Two methods of neutrophil isolation were used: a neutrophil isolation on the basis of hypotonic erythrocyte lysis (in two variants: after the erythrocyte lysis proper, the cells were centrifuged at either 200 g or 1000 g, and b neutrophil isolation with FACS Lysing Solution as the lysing agent. The viability of the isolated neutrophils was evaluated on the basis of apoptosis and necrosis. The results obtained with flow cytometry (FCM suggest that, from the isolation techniques used, the method based on FACS Lysing Solution impaired the neutrophil viability least. After the application of this method, 5.36 ± 2.15% of neutrophils were apoptotic and 0.51 ± 0.12% were necrotic. In contrast, when the hypotonic erythrocyte lysis was used, the proportion of apoptotic neutrophils amounted to 42.14 ± 7.12% and 49.00 ± 14.70%, respectively, and 41.12 ± 5.55% and 36.91 ± 24.38% respectively of necrotic neutrophils (P < 0.01. This was also confirmed by the light microscopy. After the isolation with FASC Lysing Solution, 1.92 ± 1.74% of neutrophils were apoptotic and 1.05 ± 0.76% were necrotic, as distinct from after the hypotonic erythrocyte lysis where 9.43 ± 3.69% of neutrophils were apoptotic and 12.67 ± 4.74% of necrotic after centrifugation at 200 g, while 12.60 ± 4.35 were apoptotic and 14.96 ± 12.64% were necrotic after centrifugation at 1000 g. It follows from the above-mentioned data that hypotonic lysis is not a suitable method for the isolation of neutrophils, as the method itself markedly affects cell viability.

  4. Inhibition of metastatic tumor growth in mouse lung by repeated administration of polyethylene glycol-conjugated catalase: quantitative analysis with firefly luciferase-expressing melanoma cells.

    Science.gov (United States)

    Hyoudou, Kenji; Nishikawa, Makiya; Umeyama, Yukari; Kobayashi, Yuki; Yamashita, Fumiyoshi; Hashida, Mitsuru

    2004-11-15

    To develop a novel and effective approach to inhibit tumor metastasis based on controlled delivery of catalase, we first evaluated the characteristics of the disposition and proliferation of tumor cells. Then, we examined the effects of polyethylene glycol-conjugated catalase (PEG-catalase) on tumor metastasis. On the basis of the results obtained, PEG-catalase was repetitively administered to completely suppress the growth of tumor cells. Murine melanoma B16-BL6 cells were stably transfected with firefly luciferase gene to obtain B16-BL6/Luc cells. These cells were injected intravenously into syngeneic C57BL/6 mice. PEG-catalase was injected intravenously, and the effect was evaluated by measuring the luciferase activity as the indicator of the number of tumor cells. At 1 hour after injection of B16-BL6/Luc cells, 60 to 90% of the injected cells were recovered in the lung. The numbers decreased to 2 to 4% at 24 hours, then increased. An injection of PEG-catalase just before inoculation significantly reduced the number of tumor cells at 24 hours. Injection of PEG-catalase at 1 or 3 days after inoculation was also effective in reducing the cell numbers. Daily dosing of PEG-catalase greatly inhibited the proliferation and the number assayed at 14 days after inoculation was not significantly different from the minimal number observed at 1 day, suggesting that the growth had been markedly suppressed by the treatment. These findings indicate that sustained catalase activity in the blood circulation can prevent the multiple processes of tumor metastasis in the lung, which could lead to a state of tumor dormancy.

  5. Development of a dual luciferase activity and fluorescamine protein assay adapted to a 384 micro-well plate format: Reducing variability in human luciferase transactivation cell lines aimed at endocrine active substances

    Science.gov (United States)

    Brennan, Jennifer; Tillitt, Donald E.

    2018-01-01

    There is a need to adapt cell bioassays to 384-well and 1536-well formats instead of the traditional 96-well format as high-throughput screening (HTS) demands increase. However, the sensitivity and performance of the bioassay must be re-verified in these higher micro-well plates, and verification of cell health must also be HT (high-throughput). We have adapted two commonly used human breast luciferase transactivation cell bioassays, the recently re-named estrogen agonist/antagonist screening VM7Luc4E2 cell bioassay (previously designated BG1Luc4E2) and the androgen/glucocorticoid screening MDA-kb2 cell bioassay, to 384-well formats for HTS of endocrine-active substances (EASs). This cost-saving adaptation includes a fast, accurate, and easy measurement of protein amount in each well via the fluorescamine assay with which to normalize luciferase activity of cell lysates without requiring any transfer of the cell lysates. Here we demonstrate that by accounting for protein amount in the cell lysates, antagonistic agents can easily be distinguished from cytotoxic agents in the MDA-kb2 and VM7Luc4E2 cell bioassays. Additionally, we demonstrate via the fluorescamine assay improved interpretation of luciferase activity in wells along the edge of the plate (the so-called “edge effect”), thereby increasing usable wells to the entire plate, not just interior wells.

  6. The disulfide-rich Metridia luciferase refolded from E. coli inclusion bodies reveals the properties of a native folded enzyme produced in insect cells.

    Science.gov (United States)

    Markova, Svetlana V; Larionova, Marina D; Gorbunova, Darya A; Vysotski, Eugene S

    2017-10-01

    The bioluminescence of a marine copepod Metridia longa is determined by a small secreted coelenterazine-dependent luciferase that uses coelenterazine as a substrate of enzymatic reaction to generate light (λ max =480nm). To date, four different isoforms of the luciferase differing in size, sequences, and properties have been cloned by functional screening. All of them contain ten conserved Cys residues that suggests up to five SS intramolecular bonds per luciferase molecule. Whereas the use of copepod luciferases as bioluminescent reporters in biomedical research in vivo is growing from year to year, their application for in vitro assays is still limited by the difficulty in obtaining significant amounts of luciferase. The most cost-effective host for producing recombinant proteins is Escherichia coli. However, prokaryotic and eukaryotic cells maintain the reductive environment in cytoplasm that hinders the disulfide bond formation and consequently the proper folding of luciferase. Here we report the expression of the MLuc7 isoform of M. longa luciferase in E. coli cells and the efficient procedure for refolding from inclusion bodies yielding a high-active monomeric protein. Furthermore, in a set of identical experiments we demonstrate that bioluminescent and structural features of MLuc7 produced in bacterial cells are identical to those of MLuc7 isoform produced from culture medium of insect cells. Although the yield of high-purity protein is only 6mg/L, the application of E. coli cells to produce the luciferase is simpler and more cost-effective than the use of insect cells. We expect that the suggested technology of Metridia luciferase production allows obtaining of sufficient amounts of protein both for the development of novel in vitro analytical assays with the use of MLuc7 as a label and for structural studies. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Influence of harvesting and processing methods on organic viability of soybean seed

    Directory of Open Access Journals (Sweden)

    Đukanović Lana

    2000-01-01

    Full Text Available Organic viability of soybean seed for three soybean varieties - elite (Bosa, ZPS 015 and Nena depending on methods of manipulation with seeds during harvesting and processing phase were determined in this paper. Trial was conducted in Zemun Polje during 1999; manual and mechanized harvesting or processing methods were applied. Seed germination was tested using ISTA methods (Standard method and Cold test. Following parameters were evaluated: germination viability, germination, rate-speed of emergence, length of hypocotile and main root Rate-speed of emergence was based on number of emerged plants per day. Length of hypocotile or root and percent of germination determined vigour index. Based on obtained results it maybe concluded that methods of seed manipulation during harvesting or processing phase were influenced on soybean seed quality parameters evaluated. Ways of seed manipulation - methods evaluated were influenced organic viability of soybean seed by decreasing germination viability, total germination and length of main root.

  8. Aerosol delivery of Akt controls protein translation in the lungs of dual luciferase reporter mice.

    Science.gov (United States)

    Tehrani, A M; Hwang, S-K; Kim, T-H; Cho, C-S; Hua, J; Nah, W-S; Kwon, J-T; Kim, J-S; Chang, S-H; Yu, K-N; Park, S-J; Bhandari, D R; Lee, K-H; An, G-H; Beck, G R; Cho, M-H

    2007-03-01

    Lung cancer has emerged as a leading cause of cancer death in the world; however, most of the current conventional therapies are not sufficiently effective in altering the progression of disease. Therefore, development of novel treatment approaches is needed. Although several genes and methods have been used for cancer gene therapy, a number of problems such as specificity, efficacy and toxicity reduce their application. This has led to re-emergence of aerosol gene delivery as a noninvasive method for lung cancer treatment. In this study, nano-sized glucosylated polyethyleneimine (GPEI) was used as a gene delivery carrier to investigate the effects of Akt wild type (WT) and kinase deficient (KD) on Akt-related signaling pathways and protein translation in the lungs of CMV- LucR-cMyc-IRES-LucF dual reporter mice. These mice are a powerful tool for the discrimination between cap-dependent/-independent protein translation. Aerosols containing self-assembled nano-sized GPEI/Akt WT or GPEI/Akt KD were delivered into the lungs of reporter mice through nose-only-inhalation-chamber with the aid of nebulizer. Aerosol delivery of Akt WT caused the increase of protein expression levels of Akt-related signals, whereas aerosol delivery of Akt KD did not. Furthermore, dual luciferase activity assay showed that aerosol delivery of Akt WT enhanced cap-dependent protein translation, whereas a reduction in cap-dependent protein translation by Akt KD was observed. Our results clearly showed that targeting Akt may be a good strategy for prevention as well as treatment of lung cancer. These studies suggest that our aerosol delivery is compatible for in vivo gene delivery which could be used as a noninvasive gene therapy in the future.

  9. The cybernetics of viability: an overview

    Science.gov (United States)

    Nechansky, Helmut

    2011-10-01

    A three-level approach to viability is developed, considering (1) living systems, (2) a niche, understood as the area within the reach of their actions, and (3) an environment. A systematic analysis of the interrelations between these levels shows that living systems emerge with matter/energy processing systems. These can add controller structures when producing excess energy. A three-sensor controller structure enables a living system to deal with unfavourable and scarce environments. Further evolution of these controller structures offers improved ways to act on niches. Maintaining niches in scarce environments can require technology or economy. So social systems emerge, which are understood as aggregates of living systems. Basic patterns of interactions within social systems are analysed. So the introduction of the notion of the niche into the discussion of viability allows us to explain phenomena ranging from properties of single living systems to societal organization.

  10. Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation.

    Science.gov (United States)

    Li, Jian-Feng; Bush, Jenifer; Xiong, Yan; Li, Lei; McCormack, Matthew

    2011-01-01

    Protein-protein interactions (PPIs) constitute the regulatory network that coordinates diverse cellular functions. There are growing needs in plant research for creating protein interaction maps behind complex cellular processes and at a systems biology level. However, only a few approaches have been successfully used for large-scale surveys of PPIs in plants, each having advantages and disadvantages. Here we present split firefly luciferase complementation (SFLC) as a highly sensitive and noninvasive technique for in planta PPI investigation. In this assay, the separate halves of a firefly luciferase can come into close proximity and transiently restore its catalytic activity only when their fusion partners, namely the two proteins of interest, interact with each other. This assay was conferred with quantitativeness and high throughput potential when the Arabidopsis mesophyll protoplast system and a microplate luminometer were employed for protein expression and luciferase measurement, respectively. Using the SFLC assay, we could monitor the dynamics of rapamycin-induced and ascomycin-disrupted interaction between Arabidopsis FRB and human FKBP proteins in a near real-time manner. As a proof of concept for large-scale PPI survey, we further applied the SFLC assay to testing 132 binary PPIs among 8 auxin response factors (ARFs) and 12 Aux/IAA proteins from Arabidopsis. Our results demonstrated that the SFLC assay is ideal for in vivo quantitative PPI analysis in plant cells and is particularly powerful for large-scale binary PPI screens.

  11. Large-scale protein-protein interaction analysis in Arabidopsis mesophyll protoplasts by split firefly luciferase complementation.

    Directory of Open Access Journals (Sweden)

    Jian-Feng Li

    Full Text Available Protein-protein interactions (PPIs constitute the regulatory network that coordinates diverse cellular functions. There are growing needs in plant research for creating protein interaction maps behind complex cellular processes and at a systems biology level. However, only a few approaches have been successfully used for large-scale surveys of PPIs in plants, each having advantages and disadvantages. Here we present split firefly luciferase complementation (SFLC as a highly sensitive and noninvasive technique for in planta PPI investigation. In this assay, the separate halves of a firefly luciferase can come into close proximity and transiently restore its catalytic activity only when their fusion partners, namely the two proteins of interest, interact with each other. This assay was conferred with quantitativeness and high throughput potential when the Arabidopsis mesophyll protoplast system and a microplate luminometer were employed for protein expression and luciferase measurement, respectively. Using the SFLC assay, we could monitor the dynamics of rapamycin-induced and ascomycin-disrupted interaction between Arabidopsis FRB and human FKBP proteins in a near real-time manner. As a proof of concept for large-scale PPI survey, we further applied the SFLC assay to testing 132 binary PPIs among 8 auxin response factors (ARFs and 12 Aux/IAA proteins from Arabidopsis. Our results demonstrated that the SFLC assay is ideal for in vivo quantitative PPI analysis in plant cells and is particularly powerful for large-scale binary PPI screens.

  12. Transgenic Mouse Model Harboring the Transcriptional Fusion Ccl20-Luciferase as a Novel Reporter of Pro-Inflammatory Response

    Science.gov (United States)

    Crispo, Martina; Van Maele, Laurye; Tabareau, Julien; Cayet, Delphine; Errea, Agustina; Ferreira, Ana María; Rumbo, Martin; Sirard, Jean Claude

    2013-01-01

    The chemokine CCL20, the unique ligand of CCR6 functions as an attractant of immune cells. Expression of CCL20 is induced by Toll-like Receptor (TLR) signaling or proinflammatory cytokine stimulation. However CCL20 is also constitutively produced at specific epithelial sites of mucosa. This expression profile is achieved by transcriptional regulation. In the present work we characterized regulatory features of mouse Ccl20 gene. Transcriptional fusions between the mouse Ccl20 promoter and the firefly luciferase (luc) encoding gene were constructed and assessed in in vitro and in vivo assays. We found that liver CCL20 expression and luciferase activity were upregulated by systemic administration of the TLR5 agonist flagellin. Using shRNA and dominant negative form specific for mouse TLR5, we showed that this expression was controlled by TLR5. To address in situ the regulation of gene activity, a transgenic mouse line harboring a functional Ccl20-luc fusion was generated. The luciferase expression was highly concordant with Ccl20 expression in different tissues. Our data indicate that the transgenic mouse model can be used to monitor activation of innate response in vivo. PMID:24265691

  13. Development of microLIPS (Luciferase Immunoprecipitation Systems): a novel microfluidic assay for rapid serum antibody detection

    Science.gov (United States)

    Chandrangsu, Matt; Burbelo, Peter D.; Iadarola, Michael J.; Smith, Paul D.; Morgan, Nicole Y.

    2012-06-01

    There is considerable interest in the development of rapid, point-of-care antibody detection for the diagnosis of infectious and auto-immune diseases. In this paper, we present work on the development of a self-contained microfluidic format for the Luciferase Immunoprecipitation Systems (LIPS) assay. Whereas the majority of immunoassays for antigen-specific antibodies employ either bacteria- or yeast-expressed proteins and require the use of secondary antibodies, the LIPS technique uses a fusion protein comprised of a Renilla luciferase reporter and the antigen of interest produced via mammalian cell culture, ensuring the addition of mammalian post-translational modifications. Patient serum is mixed with the fusion protein and passed over immobilized Protein A/G; after washing, the only remaining luciferase-tagged antigens are those retained by specific antibodies. These can be quantitatively measured using chemiluminescence upon the introduction of coelenterazine. The assay has been successfully employed for a wide variety of diseases in a microwell format. We report on a recent demonstration of rapid HSV-2 diagnosis with the LIPS assay in a microfluidic format, using one microliter of serum and obtaining results in under ten minutes. We will also discuss recent progress on two fronts, both aimed at the deployment of this technology in the field: first, simplifying assay operation through the automation of flow control using power-free means; and second, efforts to increase signal levels, primarily through strategies to increase antibody binding capacity, in order to move towards portable battery powered electronics.

  14. Design and introduction of a disulfide bridge in firefly luciferase: increase of thermostability and decrease of pH sensitivity.

    Science.gov (United States)

    Imani, Mehdi; Hosseinkhani, Saman; Ahmadian, Shahin; Nazari, Mahboobeh

    2010-08-01

    The thermal sensitivity and pH-sensitive spectral properties of firefly luciferase have hampered its application in a variety of fields. It is proposed that the stability of a protein can be increased by introduction of disulfide bridge that decreases the configurational entropy of unfolding. A disulfide bridge is introduced into Photinus pyralis firefly luciferase to make two separate mutant enzymes with a single bridge. Even though the A103C/S121C mutant showed remarkable thermal stability, its specific activity decreased, whereas the A296C/A326C mutant showed tremendous thermal stability, relative pH insensitivity and 7.3-fold increase of specific activity. Moreover, the bioluminescence emission spectrum of A296C/A326C was resistant against higher temperatures (37 degrees C). Far-UV CD analysis showed slight secondary structure changes for both mutants. Thermal denaturation analysis showed that conformational stabilities of A103C/S121C and A296C/A326C are more than native firefly luciferase. It is proposed that since A296 and A326 are situated in the vicinity of the enzyme active site microenvironment in comparison with A103 and S121, the formation of a disulfide bridge in this region has more impact on enzyme kinetic characteristics.

  15. Myocardial viability assessment using nuclear imaging

    International Nuclear Information System (INIS)

    Matsunari, Ichiro; Hisada, Kinichi; Taki, Junichi; Nakajima, Kenichi; Tonami, Norihisa

    2003-01-01

    Myocardial assessment continues to be an issue in patients with coronary artery disease and left ventricular dysfunction. Nuclear imaging has long played an important role in this field. In particular, PET imaging using 18 F-fluorodeoxyglucose is regarded as the metabolic gold standard of tissue viability, which has been supported by a wide clinical experience. Viability assessment using SPECT techniques has gained more wide-spread clinical acceptance than PET, because it is more widely available at lower cost. Moreover, technical advances in SPECT technology such as gated-SPECT further improve the diagnostic accuracy of the test. However, other imaging techniques such as dobutamine echocardiography have recently emerged as competitors to nuclear imaging. It is also important to note that they sometimes may work in a complementary fashion to nuclear imaging, indicating that an appropriate use of these techniques may significantly improve their overall accuracy. In keeping these circumstances in mind, further efforts are necessary to further improve the diagnostic performance of nuclear imaging as a reliable viability test. (author) 107 refs

  16. A comparison of cryopreservation methods: Slow-cooling vs. rapid-cooling based on cell viability, oxidative stress, apoptosis, and CD34+ enumeration of human umbilical cord blood mononucleated cells

    Directory of Open Access Journals (Sweden)

    Sandra Ferry

    2011-09-01

    Full Text Available Abstract Background The finding of human umbilical cord blood as one of the most likely sources of hematopoietic stem cells offers a less invasive alternative for the need of hematopoietic stem cell transplantation. Due to the once-in-a-life time chance of collecting it, an optimum cryopreservation method that can preserve the life and function of the cells contained is critically needed. Methods Until now, slow-cooling has been the routine method of cryopreservation; however, rapid-cooling offers a simple, efficient, and harmless method for preserving the life and function of the desired cells. Therefore, this study was conducted to compare the effectiveness of slow- and rapid-cooling to preserve umbilical cord blood of mononucleated cells suspected of containing hematopoietic stem cells. The parameters used in this study were differences in cell viability, malondialdehyde content, and apoptosis level. The identification of hematopoietic stem cells themselves was carried out by enumerating CD34+ in a flow cytometer. Results Our results showed that mononucleated cell viability after rapid-cooling (91.9% was significantly higher than that after slow-cooling (75.5%, with a p value = 0.003. Interestingly, the malondialdehyde level in the mononucleated cell population after rapid-cooling (56.45 μM was also significantly higher than that after slow-cooling (33.25 μM, with a p value p value = 0.138. However, CD34+ enumeration was much higher in the population that underwent slow-cooling (23.32 cell/μl than in the one that underwent rapid-cooling (2.47 cell/μl, with a p value = 0.001. Conclusions Rapid-cooling is a potential cryopreservation method to be used to preserve the umbilical cord blood of mononucleated cells, although further optimization of the number of CD34+ cells after rapid-cooling is critically needed.

  17. Financial viability of the Sonora-Baja California interconnection line

    International Nuclear Information System (INIS)

    Alonso, G.; Ortega, G.

    2017-09-01

    In the Development Program of the National Electricity Sector 2015-2029, an electric interconnection line between Sonora and Baja California (Mexico) is proposed, this study analyzes the financial viability of this interconnection line based on the maximum hourly and seasonal energy demand between both regions and proposes alternatives for the supply of electric power that supports the economic convenience of this interconnection line. The results show that additional capacity is required in Sonora to cover the maximum demands of both regions since in the current condition of the National Electric System the interconnection line is not justified. (Author)

  18. The effects of storage conditions on the viability of ...

    African Journals Online (AJOL)

    SARAKA DANIEL

    2015-01-07

    Jan 7, 2015 ... E-mail: ndsaraka@yahoo.fr, ndsaraka@gmail.com. Tel: 225 ..... sample viability and control of contaminants that may mask the ... viability and composition of the Escherichia coli flora in faecal ... Microbiologie alimentaire, 8e.

  19. Pollen diversity, viability and floral structure of some Musa genotypes

    African Journals Online (AJOL)

    Pollen diversity, viability and floral structure of some Musa genotypes. ... This experiment was designed to study the floral structure, pollen morphology and the potential pollen viability of five Musa genotypes obtained ... HOW TO USE AJOL.

  20. Noninvasive Real-Time Assessment of Cell Viability in a Three-Dimensional Tissue.

    Science.gov (United States)

    Mahfouzi, Seyed Hossein; Amoabediny, Ghassem; Doryab, Ali; Safiabadi-Tali, Seyed Hamid; Ghanei, Mostafa

    2018-04-01

    Maintaining cell viability within 3D tissue engineering scaffolds is an essential step toward a functional tissue or organ. Assessment of cell viability in 3D scaffolds is necessary to control and optimize tissue culture process. Monitoring systems based on respiration activity of cells (e.g., oxygen consumption) have been used in various cell cultures. In this research, an online monitoring system based on respiration activity was developed to monitor cell viability within acellular lung scaffolds. First, acellular lung scaffolds were recellularized with human umbilical cord vein endothelial cells, and then, cell viability was monitored during a 5-day period. The real-time monitoring system generated a cell growth profile representing invaluable information on cell viability and proliferative states during the culture period. The cell growth profile obtained by the monitoring system was consistent with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis and glucose consumption measurement. This system provided a means for noninvasive, real-time, and repetitive investigation of cell viability. Also, we showed the applicability of this monitoring system by introducing shaking as an operating parameter in a long-term culture.

  1. The examination of urine samples for pathogenic microbes by the luciferase assay for ATP. 1: The effect of the presence of fungi, fungal like bacteria and kidney cells in urine samples

    Science.gov (United States)

    Bush, V. N.

    1973-01-01

    A method for accurately determining urinary tract infections in man is introduced. The method is based on adenosine triphosphate (ATP) concentration in urine samples after removing nonbacterial ATP. Adenosine triphosphate concentration is measured from the bioluminescent reaction of luciferase when mixed with ATP. An examination was also made of the effectiveness of rupturing agents on monkey kidney cells Candia albicans, a Rhodotorula species, and a Streptomyces species in determining whether these cells could contribute ATP to the bacterial ATP value of a urine sample.

  2. Echinococcus metacestode: in search of viability markers.

    Science.gov (United States)

    Gottstein, Bruno; Wang, Junhua; Blagosklonov, Oleg; Grenouillet, Frédéric; Millon, Laurence; Vuitton, Dominique A; Müller, Norbert

    2014-01-01

    Epidemiological studies have demonstrated that most humans infected with Echinococcus spp. exhibit resistance to disease. When infection leads to disease, the parasite is partially controlled by host immunity: in case of immunocompetence, the normal alveolar echinococcosis (AE) or cystic echinococcosis (CE) situation, the metacestode grows slowly, and first clinical signs appear years after infection; in case of impaired immunity (AIDS; other immunodeficiencies), uncontrolled proliferation of the metacestode leads to rapidly progressing disease. Assessing Echinococcus multilocularis viability in vivo following therapeutic interventions in AE patients may be of tremendous benefit when compared with the invasive procedures used to perform biopsies. Current options are F18-fluorodeoxyglucose-positron emission tomography (FDG-PET), which visualizes periparasitic inflammation due to the metabolic activity of the metacestode, and measurement of antibodies against recEm18, a viability-associated protein, that rapidly regresses upon metacestode inactivation. For Echinococcus granulosus, similar prognosis-associated follow-up parameters are still lacking but a few candidates may be listed. Other possible markers include functional and diffusion-weighted Magnetic Resonance Imaging (MRI), and measurement of products from the parasite (circulating antigens or DNA), and from the host (inflammation markers, cytokines, or chemokines). Even though some of them have been promising in pilot studies, none has been properly validated in an appropriate number of patients until now to be recommended for further use in clinical settings. There is therefore still a need to develop reliable tools for improved viability assessment to provide the sufficient information needed to reliably withdraw anti-parasite benzimidazole chemotherapy, and a basis for the development of new alternative therapeutic tools. © B. Gottstein et al., published by EDP Sciences, 2014.

  3. Comparing the effects of tetrabromobisphenol-A, bisphenol A, and their potential replacement alternatives, TBBPA-bis(2,3-dibromopropyl ether) and bisphenol S, on cell viability and messenger ribonucleic acid expression in chicken embryonic hepatocytes.

    Science.gov (United States)

    Ma, Melissa; Crump, Doug; Farmahin, Reza; Kennedy, Sean W

    2015-02-01

    A market for alternative brominated flame retardants (BFRs) has emerged recently due to the phase out of persistent and inherently toxic BFRs. Several of these replacement compounds have been detected in environmental matrices, including wild birds. A chicken embryonic hepatocyte (CEH) assay was utilized to assess the effects of the BFR, tetrabromobisphenol-A (TBBPA), and its replacement alternative, tetrabromobisphenol A bis(2,3-dibromopropyl ether [TBBPA-DBPE]) on cell viability and messenger ribonucleic acid (mRNA) expression. Bisphenol A (BPA) and 1 of its replacement alternatives, bisphenol S (BPS), were also screened for effects. Both TBBPA and BPA decreased CEH viability with calculated median lethal concentration (LC50) values of 40.6 μM and 61.7 μM, respectively. However, the replacement alternatives, TBBPA-DBPE and BPS, did not affect cell viability (up to 300 μM). Effects on mRNA expression were determined using an Avian ToxChip polymerse chain reaction (PCR) array and a real-time (RT)-PCR assay for the estrogen-responsive genes, apolipoproteinII (ApoII) and vitellogenin (Vtg). A luciferase reporter gene assay was used to assess dioxin-like effects. Tetrabromobisphenol-A altered mRNA levels of 4 genes from multiple toxicity pathways and increased luciferase activity in the luciferase reporter gene assay, whereas its alternative, TBBPA-DBPE, only altered 1 gene on the array, Cyp1a4, and increased luciferase activity. At 300 μM, a concentration that decreased cell viability for TBBPA and BPA, the BPA replacement, BPS, altered the greatest number of transcripts, including both ApoII and Vtg. Bisphenol A exposure did not alter any genes on the array but did up-regulate Vtg at 10 μM. Characterization of the potential toxicological and molecular-level effects of these compounds will ideally be useful to chemical regulators tasked with assessing the risk of new and existing chemicals. © 2014 SETAC.

  4. Adenovirus-assisted lipofection: efficient in vitro gene transfer of luciferase and cytosine deaminase to human smooth muscle cells.

    Science.gov (United States)

    Kreuzer, J; Denger, S; Reifers, F; Beisel, C; Haack, K; Gebert, J; Kübler, W

    1996-07-01

    Smooth muscle cells (SMC) are a central cell type involved in multiple processes of coronary artery diseases including restenosis and therefore are major target cells for different aspects of gene transfer. Previous attempts to transfect primary arterial cells using different techniques like liposomes, CaPO4 and electroporation resulted in only low transfection efficiency. The development of recombinant adenoviruses dramatically improved the delivery of foreign genes into different cell types including SMC. However, cloning and identification of recombinants remain difficult and time-consuming techniques. The present study demonstrates that a complex consisting of reporter plasmid encoding firefly luciferase (pLUC), polycationic liposomes and replication-deficient adenovirus was able to yield very high in vitro transfection of primary human smooth muscle cells under optimized conditions. The technique of adenovirus-assisted lipofection (AAL) increases transfer and expression of plasmid DNA in human smooth muscle cells in vitro up to 1000-fold compared to lipofection. To verify the applicability of AAL for gene transfer into human smooth muscle cells we studied a gene therapy approach to suppress proliferation of SMC in vitro, using the prokaryotic cytosine deaminase gene (CD) which enables transfected mammalian cells to deaminate 5-fluorocytosine (5-FC) to the highly toxic 5-fluorouracil (5-FU). The effect of a transient CD expression on RNA synthesis was investigated by means of a cotransfection with a RSV-CD expression plasmid and the luciferase reporter plasmid. Western blot analysis demonstrated high expression of CD protein in transfected SMC. Cotransfected SMC demonstrated two-fold less luciferase activity in the presence of 5-FC (5 mmol/l) after 48 h compared to cells transfected with a non-CD coding plasmid. The data demonstrate that a transient expression of CD could be sufficient to reduce the capacity of protein synthesis in human SMC. This simple and

  5. A dual-color luciferase assay system reveals circadian resetting of cultured fibroblasts by co-cultured adrenal glands.

    Directory of Open Access Journals (Sweden)

    Takako Noguchi

    Full Text Available In mammals, circadian rhythms of various organs and tissues are synchronized by pacemaker neurons in the suprachiasmatic nucleus (SCN of the hypothalamus. Glucocorticoids released from the adrenal glands can synchronize circadian rhythms in other tissues. Many hormones show circadian rhythms in their plasma concentrations; however, whether organs outside the SCN can serve as master synchronizers to entrain circadian rhythms in target tissues is not well understood. To further delineate the function of the adrenal glands and the interactions of circadian rhythms in putative master synchronizing organs and their target tissues, here we report a simple co-culture system using a dual-color luciferase assay to monitor circadian rhythms separately in various explanted tissues and fibroblasts. In this system, circadian rhythms of organs and target cells were simultaneously tracked by the green-emitting beetle luciferase from Pyrearinus termitilluminans (ELuc and the red-emitting beetle luciferase from Phrixothrix hirtus (SLR, respectively. We obtained tissues from the adrenal glands, thyroid glands, and lungs of transgenic mice that expressed ELuc under control of the promoter from a canonical clock gene, mBmal1. The tissues were co-cultured with Rat-1 fibroblasts as representative target cells expressing SLR under control of the mBmal1 promoter. Amplitudes of the circadian rhythms of Rat-1 fibroblasts were potentiated when the fibroblasts were co-cultured with adrenal gland tissue, but not when co-cultured with thyroid gland or lung tissue. The phases of Rat-1 fibroblasts were reset by application of adrenal gland tissue, whereas the phases of adrenal gland tissue were not influenced by Rat-1 fibroblasts. Furthermore, the effect of the adrenal gland tissue on the fibroblasts was blocked by application of a glucocorticoid receptor (GR antagonist. These results demonstrate that glucocorticoids are strong circadian synchronizers for fibroblasts and that

  6. Myocardial Viability on Cardiac Magnetic Resonance.

    Science.gov (United States)

    Souto, Ana Luiza Mansur; Souto, Rafael Mansur; Teixeira, Isabella Cristina Resende; Nacif, Marcelo Souto

    2017-05-01

    The study of myocardial viability is of great importance in the orientation and management of patients requiring myocardial revascularization or angioplasty. The technique of delayed enhancement (DE) is accurate and has transformed the study of viability into an easy test, not only for the detection of fibrosis but also as a binary test detecting what is viable or not. On DE, fibrosis equal to or greater than 50% of the segmental area is considered as non-viable, whereas that below 50% is considered viable. During the same evaluation, cardiac magnetic resonance (CMR) may also use other techniques for functional and perfusion studies to obtain a global evaluation of ischemic heart disease. This study aims to highlight the current concepts and broadly emphasize the use of CMR as a method that over the last 20 years has become a reference in the detection of infarction and assessment of myocardial viability. Resumo O estudo de viabilidade miocárdica é de grande importância para a orientação e manejo de pacientes que necessitam de cirurgia de revascularização miocárdica ou angioplastia. A técnica de realce tardio (RT) é precisa e transformou o estudo de viabilidade em um teste fácil, não só para a detecção de fibrose, mas também como um modelo binário para a detecção do que é ou não é viável. Uma fibrose identificada pelo RT é considerada como não viável quando igual ou maior do que 50% da área segmentar e como viável quando menor que 50%. A ressonância magnética cardíaca (RMC) também pode lançar mão de outras técnicas para estudo funcional e de perfusão para uma avaliação global da doença isquêmica do coração no mesmo exame. Este estudo tem como objetivo destacar os conceitos atuais e enfatizar amplamente o uso da RMC como um método que nos últimos 20 anos se tornou referência na detecção de infarto e avaliação de viabilidade miocárdica.

  7. Gene silencing in primary and metastatic tumors by small interfering RNA delivery in mice: quantitative analysis using melanoma cells expressing firefly and sea pansy luciferases.

    Science.gov (United States)

    Takahashi, Yuki; Nishikawa, Makiya; Kobayashi, Naoki; Takakura, Yoshinobu

    2005-07-20

    Silencing of oncogenes or other genes contributing to tumor malignancy or progression by RNA interference (RNAi) offers a promising approach to treating tumor patients. To achieve RNAi-based tumor therapy, a small interfering RNA (siRNA) or siRNA-expressing vector needs to be delivered to tumor cells, but little information about its in vivo delivery has been reported. In this study, we examined whether the expression of the target gene in tumor cells can be suppressed by the delivery of RNAi effectors to primary and metastatic tumor cells. To quantitatively evaluate the RNAi effects in tumor cells, mouse melanoma B16-BL6 cells were stably transfected with both firefly (a model target gene) and sea pansy (an internal standard gene) luciferase genes to obtain B16-BL6/dual Luc cells. The target gene expression in subcutaneous primary tumors of B16-BL6/dual Luc cells was significantly suppressed by direct injection of the RNAi effectors followed by electroporation. The expression in metastatic hepatic tumors was also significantly reduced by an intravenous injection of either RNAi effector by the hydrodynamics-based procedure. These results indicate that the both RNAi effectors have a potential to silence target gene in tumor cells in vivo when successfully delivered to tumor cells.

  8. Exercise regulates breast cancer cell viability

    DEFF Research Database (Denmark)

    Dethlefsen, Christine; Lillelund, Christian; Midtgaard, Julie

    2016-01-01

    .003) and cytokines. Yet, these systemic adaptations had no effect on breast cancer cell viability in vitro. During 2 h of acute exercise, increases in serum lactate (6-fold, p ... no impact. Our data question the prevailing dogma that training-dependent baseline reductions in risk factors mediate the protective effect of exercise on breast cancer. Instead, we propose that the cancer protection is driven by accumulative effects of repeated acute exercise responses.......Purpose: Exercise decreases breast cancer risk and disease recurrence, but the underlying mechanisms are unknown. Training adaptations in systemic factors have been suggested as mediating causes. We aimed to examine if systemic adaptations to training over time, or acute exercise responses...

  9. Crystal structure of the bacterial luciferase/flavin complex provides insight into the function of the beta subunit.

    Science.gov (United States)

    Campbell, Zachary T; Weichsel, Andrzej; Montfort, William R; Baldwin, Thomas O

    2009-07-07

    Bacterial luciferase from Vibrio harveyi is a heterodimer composed of a catalytic alpha subunit and a homologous but noncatalytic beta subunit. Despite decades of enzymological investigation, structural evidence defining the active center has been elusive. We report here the crystal structure of V. harveyi luciferase bound to flavin mononucleotide (FMN) at 2.3 A. The isoalloxazine ring is coordinated by an unusual cis-Ala-Ala peptide bond. The reactive sulfhydryl group of Cys106 projects toward position C-4a, the site of flavin oxygenation. This structure also provides the first data specifying the conformations of a mobile loop that is crystallographically disordered in both prior crystal structures [(1995) Biochemistry 34, 6581-6586; (1996) J. Biol. Chem. 271, 21956 21968]. This loop appears to be a boundary between solvent and the active center. Within this portion of the protein, a single contact was observed between Phe272 of the alpha subunit, not seen in the previous structures, and Tyr151 of the beta subunit. Substitutions at position 151 on the beta subunit caused reductions in activity and total quantum yield. Several of these mutants were found to have decreased affinity for reduced flavin mononucleotide (FMNH(2)). These findings partially address the long-standing question of how the beta subunit stabilizes the active conformation of the alpha subunit, thereby participating in the catalytic mechanism.

  10. Advances and Challenges in Viability Detection of Foodborne Pathogens

    Directory of Open Access Journals (Sweden)

    Dexin Zeng

    2016-11-01

    Full Text Available Foodborne outbreaks are a serious public health and food safety concern worldwide. There is a great demand for rapid, sensitive, specific, and accurate methods to detect microbial pathogens in foods. Conventional methods based on cultivation of pathogens have been the gold standard protocols; however, they take up to a week to complete. Molecular assays such as polymerase chain reaction (PCR, sequencing, microarray technologies have been widely used in detection of foodborne pathogens. Among molecular assays, PCR technology conventional and real-time PCR (qPCR is most commonly used in the foodborne pathogen detection because of its high sensitivity and specificity. However, a major drawback of PCR is its inability to differentiate the DNA from dead and viable cells, and this is a critical factor for the food industry, regulatory agencies and the consumer. To remedy this shortcoming, researchers have used biological dyes such as ethidium monoazide (EMA and propidium monoazide (PMA to pretreat samples before DNA extraction to intercalate the DNA of dead cells in food samples, and then proceed with regular DNA preparation and qPCR. By combining PMA treatment with qPCR (PMA-qPCR, scientists have applied this technology to detect viable cells of various bacterial pathogens in foods. The incorporation of PMA into PCR-based assays for viability detection of pathogens in foods has increased significantly in the last decade. On the other hand, some downsides with this approach have been noted, particularly to achieve complete suppression of signal of DNA from the dead cells present in some particular food matrix. Nowadays, there is a tendency of more and more researchers adapting this approach for viability detection; and a few commercial kits based on PMA are available in the market. As time goes on, more scientists apply this approach to a broader range of pathogen detections, this viability approach (PMA or other chemicals such as platinum compound

  11. Engineering the metal sensitive sites in Macrolampis sp2 firefly luciferase and use as a novel bioluminescent ratiometric biosensor for heavy metals.

    Science.gov (United States)

    Gabriel, Gabriele V M; Viviani, Vadim R

    2016-12-01

    Most luminescent biosensors for heavy metals are fluorescent and rely on intensity measurements, whereas a few are ratiometric and rely on spectral changes. Bioluminescent biosensors for heavy metals are less common. Firefly luciferases have been coupled to responsive promoters for mercury and arsenium, and used as light on biosensors. Firefly luciferase bioluminescence spectrum is naturally sensitive to heavy metal cations such as zinc and mercury and to pH. Although pH sensitivity of firefly luciferases was shown to be useful for ratiometric estimation of intracellular pH, its potential use for ratiometric estimation of heavy metals was never considered. Using the yellow-emitting Macrolampis sp2 firefly luciferase and site-directed mutagenesis, we show that the residues H310 and E354 constitute two critical sites for metal sensitivity that can be engineered to increase sensitivity to zinc, nickel, and mercury. A linear relationship between cation concentration and the ratio of bioluminescence intensities at 550 and 610 nm allowed, for the first time, the ratiometric estimation of heavy metals concentrations down to 0.10 mM, demonstrating the potential applicability of firefly luciferases as enzymatic and intracellular ratiometric metal biosensors.

  12. Viability of Event Management Business in Batangas City, Philippine: Basis for Business Operation Initiatives

    Directory of Open Access Journals (Sweden)

    Jeninah Christia D. Borbon

    2016-11-01

    Full Text Available The research study on Viability of Event Management Business in Batangas City: Basis for Business Operation Initiatives aimed to assess the viability of this type of business using Thompson’s (2005 Dimension of Business Viability as its tool in order to create business operation initiatives. It provided a good framework for defining success factors in entrepreneurial operation initiatives in a specific business type – event management. This study utilized event organizers based in Batangas, a southern popular province, which also is a great popular destination for many types of events. Findings showed that the event management business in Batangas City is generally a personal event type of business whose year of operation ranges from one to three years, mostly link to church or reception venues and usually offers on the day coordination. In the assessment of its perceived viability, it was found out that this type of business is moderately viable in terms of market, technical, business model, management model, economic and financial, and exit strategy. Among all the dimensions tested, only market, management model, economic and financial, and exit strategy showed significant relationship with the profile variables of the event management business. From the enumerated problems encountered, those that got the highest rate were demanding clients, overbooking of reservation/exceeding number of guests and failure to meet spectators and/or competitors expectations. And, the recommended business operation initiatives were based on the weaknesses discovered using Thompson’s Dimension of Business Viability Model.

  13. Reconstruction of boundary conditions from internal conditions using viability theory

    KAUST Repository

    Hofleitner, Aude; Claudel, Christian G.; Bayen, Alexandre M.

    2012-01-01

    This article presents a method for reconstructing downstream boundary conditions to a HamiltonJacobi partial differential equation for which initial and upstream boundary conditions are prescribed as piecewise affine functions and an internal condition is prescribed as an affine function. Based on viability theory, we reconstruct the downstream boundary condition such that the solution of the Hamilton-Jacobi equation with the prescribed initial and upstream conditions and reconstructed downstream boundary condition satisfies the internal value condition. This work has important applications for estimation in flow networks with unknown capacity reductions. It is applied to urban traffic, to reconstruct signal timings and temporary capacity reductions at intersections, using Lagrangian sensing such as GPS devices onboard vehicles.

  14. Reconstruction of boundary conditions from internal conditions using viability theory

    KAUST Repository

    Hofleitner, Aude

    2012-06-01

    This article presents a method for reconstructing downstream boundary conditions to a HamiltonJacobi partial differential equation for which initial and upstream boundary conditions are prescribed as piecewise affine functions and an internal condition is prescribed as an affine function. Based on viability theory, we reconstruct the downstream boundary condition such that the solution of the Hamilton-Jacobi equation with the prescribed initial and upstream conditions and reconstructed downstream boundary condition satisfies the internal value condition. This work has important applications for estimation in flow networks with unknown capacity reductions. It is applied to urban traffic, to reconstruct signal timings and temporary capacity reductions at intersections, using Lagrangian sensing such as GPS devices onboard vehicles.

  15. Development of functional genomic tools in trematodes: RNA interference and luciferase reporter gene activity in Fasciola hepatica.

    Directory of Open Access Journals (Sweden)

    Gabriel Rinaldi

    2008-07-01

    Full Text Available The growing availability of sequence information from diverse parasites through genomic and transcriptomic projects offer new opportunities for the identification of key mediators in the parasite-host interaction. Functional genomics approaches and methods for the manipulation of genes are essential tools for deciphering the roles of genes and to identify new intervention targets in parasites. Exciting advances in functional genomics for parasitic helminths are starting to occur, with transgene expression and RNA interference (RNAi reported in several species of nematodes, but the area is still in its infancy in flatworms, with reports in just three species. While advancing in model organisms, there is a need to rapidly extend these technologies to other parasites responsible for several chronic diseases of humans and cattle. In order to extend these approaches to less well studied parasitic worms, we developed a test method for the presence of a viable RNAi pathway by silencing the exogenous reporter gene, firefly luciferase (fLUC. We established the method in the human blood fluke Schistosoma mansoni and then confirmed its utility in the liver fluke Fasciola hepatica. We transformed newly excysted juveniles of F. hepatica by electroporation with mRNA of fLUC and three hours later were able to detect luciferase enzyme activity, concentrated mainly in the digestive ceca. Subsequently, we tested the presence of an active RNAi pathway in F. hepatica by knocking down the exogenous luciferase activity by introduction into the transformed parasites of double-stranded RNA (dsRNA specific for fLUC. In addition, we tested the RNAi pathway targeting an endogenous F. hepatica gene encoding leucine aminopeptidase (FhLAP, and observed a significant reduction in specific mRNA levels. In summary, these studies demonstrated the utility of RNAi targeting reporter fLUC as a reporter gene assay to establish the presence of an intact RNAi pathway in helminth

  16. Human CRF2 α and β splice variants: pharmacological characterization using radioligand binding and a luciferase gene expression assay

    International Nuclear Information System (INIS)

    Ardati, A.; Goetschy, V.; Gottowick, J.; Henriot, S.; Deuschle, U.; Kilpatrick, G.J.; Valdenaire, O.

    1999-01-01

    Corticotropin releasing factor (CRF) receptors belong to the super-family of G protein-coupled receptors. These receptors are classified into two subtypes (CRF 1 and CRF 2 ). Both receptors are positively coupled to adenylyl cyclase but they have a distinct pharmacology and distribution in brain. Two isoforms belonging to the CRF 2 subtype receptors, CRF 2α and CRF 2β , have been identified in rat and man. The neuropeptides CRF and urocortin mediate their actions through this CRF G protein-coupled receptor family. In this report, we describe the pharmacological characterization of the recently identified hCRF 2β receptor. We have used radioligand binding with [ 125 I]-tyr 0 -sauvagine and a gene expression assay in which the firefly luciferase gene expression is under the control of cAMP responsive elements. Association kinetics of [ 125 I]-tyr 0 -sauvagine binding to the hCRF 2β receptor were monophasic while dissociation kinetics were biphasic, in agreement with the kinetics results obtained with the hCRF 2α receptor. Saturation binding analysis revealed two affinity states in HEK 293 cells with binding parameters in accord with those determined kinetically and with parameters obtained with the hCRF 2α receptor. A non-hydrolysable GTP analog, Gpp(NH)p, reduced the high affinity binding of [ 125 I]-tyr 0 -sauvagine to both hCRF 2 receptor isoforms in a similar manner. The rank order of potency of CRF agonist peptides in competition experiments was identical for both hCRF 2 α-helical CRF (9-41) oCRF). Similarly, agonist potency was similar for the two isoforms when studied using the luciferase gene reporter system. The peptide antagonist α-helical CRF (9-41) exhibited a non-competitive antagonism of urocortin-stimulated luciferase expression with both hCRF 2 receptor isoforms. Taken together, these results indicate that the pharmacological profiles of the CRF 2 splice variants are identical. This indicates that the region of the N-terminus that varies

  17. Viability of fuel cells for car production

    Energy Technology Data Exchange (ETDEWEB)

    Buchel, J.-P. [Renault, Trappes (France); Lisse, J.-P. [P.S.A., Trappes (France); Bernard, S. [Alten, Trappes (France)

    2000-07-01

    The two French car manufacturers PSA Peugeot Citroen and Renault both sell pure electric cars in an effort to reduce pollutants and carbon dioxide emissions. In addition, they have each studied fuel cell car prototypes in relation to the FEVER program for Renault and the HYDRO-GEN program for PSA. In 1999, the two manufacturers joined forces in a common program to evaluate the technical, economical and environmental viability of the fuel cell vehicle potential. The joint program has active contributions by Air Liquid, the French Atomic Energy Agency, De Nora Fuel Cells, Elf-Antar-France, Totalfina and Valeo. This paper highlighted many of the components of this program and the suitability of this new technology for industrial production at a cost competitive price. Certain automotive constraints have to be considered to propose vehicles which could provide good performance in varying temperature and operating conditions. Safety is also an important concern given that the vehicles are powered by hydrogen and a high voltage power source. Another challenges is the choice of the fuel and the economic cost of a new refueling infrastructure. Recycling was suggested as a means to recover expensive fuel cell system components such as precious catalysts, bipolar plates, membranes and other main specific parts of the fuel cell vehicle. This paper also discussed issues regarding the thermal management of the fuel cell power plant and air conditioning of the vehicles. figs.

  18. CONSERVATION OF THE VIABILITY AND VIGOR OF Araucaria angustifolia (Bert. O. Kuntze SEEDS DURING THE STORAGE

    Directory of Open Access Journals (Sweden)

    Cristhyane Garcia

    2014-12-01

    Full Text Available The conservation of Araucaria seeds is widely compromised in function of their recalcitrant feature, which hampers the planning of recovery actions of the degraded populations. Therefore, the objective of this study was to monitor the physiological changes in Araucaria seeds under controlled storage conditions, in order to get insights as to the viability and vigor conservation. The physiological quality of freshly harvested seeds was evaluated and every 60 days throughout the 180 days-storage period in laboratory ambient without thermal control, refrigerator (5 ° C, and freezer (-18 ° C until the final period of 180 days. After each sampling period, the seed viability (germination and tetrazolium tests and vigor (artificial aging, germination speed index – IVG and electrical conductivity were assessed. A reduction in the normal seedlings percentage was noticed over the period of storage of Araucaria seeds. The conservation in freezer and the lack of thermal control caused a complete loss of the seed viability at 60 and 180 days of storage, respectively. However, the refrigerator storage promoted the conservation of seed viability, with 64% germination after 180 days of storage, an event associated with the reduction of the metabolic activity of seeds. Based on the viability and vigor tests, it was concluded that storage in refrigerator provided longer storage periods to Araucaria seeds in comparison to the other storage conditions herein studied.

  19. Towards an understanding of the influence of national culture on organisational viability: An exploratory study

    Directory of Open Access Journals (Sweden)

    Awuzie Bankole O.

    2017-06-01

    Full Text Available ABSTRACT Viability connotes a system’s ability to become ultra-stable through effective self-regulation of its internal processes and information processing among its subsystems. Applying this to an infrastructure delivery system (IDS context, this study proposes that an IDS can successfully deliver on client requirements only if they attain and maintain viability. Research into the influence of National Culture (NC on an IDS’s viability appears to be lacking; hence this study. Adopting a multi-case study, qualitative research design, this study explores three IDSs involved in the delivery of infrastructure projects in two different NC contexts; Nigeria and the United Kingdom. 25 semi-structured interviews were conducted across the cases to provide for an in-depth understanding of existing interactions between participants in these delivery systems: client/project sponsor; main contractor and sub-contractors and to understand the influence of the prevailing national culture on such interactions, if any. Findings indicate that NC in project delivery environments influence the ability of IDSs to attain viability, especially as it pertains to the sustenance of Team Quality Attributes (TWQ within the system. Based on these findings, it is expected that in modelling IDSs for viability, adequate consideration should be given to the prevailing NC by project managers and planners.

  20. Effects of hydrostatic pressure and supercritical carbon dioxide on the viability of Botryococcus braunii algae cells.

    Science.gov (United States)

    Yildiz-Ozturk, Ece; Ilhan-Ayisigi, Esra; Togtema, Arnoud; Gouveia, Joao; Yesil-Celiktas, Ozlem

    2018-05-01

    In bio-based industries, Botryococcus braunii is identified as a potential resource for production of hydrocarbons having a wide range of applications in chemical and biopolymer industries. For a sustainable production platform, the algae cultivation should be integrated with downstream processes. Ideally the algae are not harvested, but the product is isolated while cultivation and growth is continued especially if the doubling time is slow. Consequently, hydrocarbons can be extracted while keeping the algae viable. In this study, the effects of pressure on the viability of B. braunii cells were tested hydrostatically and under supercritical CO 2 conditions. Viability was determined by light microscopy, methylene blue uptake and by re-cultivation of the algae after treatments to follow the growth. It was concluded that supercritical CO 2 was lethal to the algae, whereas hydrostatic pressure treatments up to 150 bar have not affected cell viability and recultivation was successful. Copyright © 2018 Elsevier Ltd. All rights reserved.

  1. In Vivo Determination of Vitamin D Function Using Transgenic Mice Carrying a Human Osteocalcin Luciferase Reporter Gene

    Directory of Open Access Journals (Sweden)

    Tomoko Nakanishi

    2013-01-01

    Full Text Available Vitamin D is an essential factor for ossification, and its deficiency causes rickets. Osteocalcin, which is a noncollagenous protein found in bone matrix and involved in mineralization and calcium ion homeostasis, is one of the major bone morphogenetic markers and is used in the evaluation of osteoblast maturation and osteogenic activation. We established transgenic mouse line expressing luciferase under the control of a 10-kb osteocalcin enhancer/promoter sequence. Using these transgenic mice, we evaluated the active forms of vitamins D2 and D3 for their bone morphogenetic function by in vivo bioluminescence. As the result, strong activity for ossification was observed with 1α,25-hydroxyvitamin D3. Our mouse system can offer a feasible detection method for assessment of osteogenic activity in the development of functional foods and medicines by noninvasive screening.

  2. Constant pH Accelerated Molecular Dynamics Investigation of the pH Regulation Mechanism of Dinoflagellate Luciferase.

    Science.gov (United States)

    Donnan, Patrick H; Ngo, Phong D; Mansoorabadi, Steven O

    2018-01-23

    The bioluminescence reaction in dinoflagellates involves the oxidation of an open-chain tetrapyrrole by the enzyme dinoflagellate luciferase (LCF). The activity of LCF is tightly regulated by pH, where the enzyme is essentially inactive at pH ∼8 and optimally active at pH ∼6. Little is known about the mechanism of LCF or the structure of the active form of the enzyme, although it has been proposed that several intramolecularly conserved histidine residues in the N-terminal region are important for the pH regulation mechanism. Here, constant pH accelerated molecular dynamics was employed to gain insight into the conformational activation of LCF induced by acidification.

  3. A rapid method for the determination of microbial susceptibility using the firefly luciferase assay for adenosine triphosphate (ATP)

    Science.gov (United States)

    Vellend, H.; Tuttle, S. A.; Barza, M.; Weinstein, L.; Picciolo, G. L.; Chappelle, E. W.

    1975-01-01

    Luciferase assay for adenosine triphosphate (ATP) was optimized for pure bacteria in broth in order to evaluate if changes in bacterial ATP content could be used as a rapid measure of antibiotic effect on microorganisms. Broth cultures of log phase bacteria were incubated at 310 K (37 C) for 2.5 hours at antimicrobial concentrations which resulted in the best discrimination between sensitive and resistant strains. Eighty-seven strains of 11 bacterial species were studied for their susceptibility to 12 commonly used antimicrobial agents: ampicillin, Penicillin G, nafcillin, carbenicillin, cephalothin, tetracycline, erythromycin, clindamycin, gentamicin, nitrofurantoin, colistin, and chloramplenicol. The major advantage of the ATP system over existing methods of rapid microbial susceptibility testing is that the assay can be made specific for bacterial ATP.

  4. Pollen viability and germination in Jatropha ribifolia and Jatropha ...

    African Journals Online (AJOL)

    The aim of this work is to assess pollen viability using the staining technique and in vitro germination with different concentrations of sucrose in Jatropha ribifolia and Jatropha mollissima, contributing to the knowledge of the reproductive biology and subsidizing their conservation, management and utilization. Pollen viability ...

  5. Sperm viability staining in ecology and evolution: potential pitfalls

    DEFF Research Database (Denmark)

    Holman, Luke

    2009-01-01

    The causes and consequences of variation in sperm quality, survival and ageing are active areas of research in ecology and evolution. In order to address these topics, many recent studies have measured sperm viability using fluorescent staining. Although sperm viability staining has produced a nu...

  6. Viability of dielectrophoretically trapped neuronal cortical cells in culture

    NARCIS (Netherlands)

    Heida, Tjitske; Vulto, P; Rutten, Wim; Marani, Enrico

    2001-01-01

    Negative dielectrophoretic trapping of neural cells is an efficient way to position neural cells on the electrode sites of planar micro-electrode arrays. The preservation of viability of the neural cells is essential for this approach. This study investigates the viability of postnatal cortical rat

  7. Flow cytometric assessment of viability of lactic acid bacteria

    NARCIS (Netherlands)

    Bunthof, C.J.; Bloemen, K.; Breeuwer, P.; Rombouts, F.M.; Abee, T.

    2001-01-01

    The viability of lactic acid bacteria is crucial for their applications as dairy starters and as probiotics. We investigated the usefulness of flow cytometry (FCM) for viability assessment of lactic acid bacteria. The esterase substrate carboxyfluorescein diacetate (cFDA) and the dye exclusion DNA

  8. Availability of elements for heterogeneous catalysis: Predicting the industrial viability of novel catalysts

    DEFF Research Database (Denmark)

    Laursen, Anders Bo; Sehested, Jens; Chorkendorff, Ib

    2018-01-01

    Growing concern regarding the sustainability of the chemical industry has driven the development of more efficient catalytic reactions. First-generation estimates of catalyst viability are based on crustal abundance, which has severe limitations. Herein, we propose a second-generation approach to...

  9. Fluorescence techniques to detect and to assess viability of plant pathogenic bacteria

    NARCIS (Netherlands)

    Chitarra, L.G.

    2001-01-01

    Plant pathogenic bacteria cause major economic losses in commercial crop production worldwide every year. The current methods used to detect and to assess the viability of bacterial pathogens and to test seed lots or plants for contamination are usually based on plate assays or on

  10. Pollen Viability and Autogamy Fitness in Bauhinia forficata Link (Fabaceae

    Directory of Open Access Journals (Sweden)

    Luana Camila Capitani

    2018-05-01

    Full Text Available ABSTRACT Bauhinia forficata (Fabaceae occurs in many phytophysiognomies of southern Brazil, however its ecological relevance is not well understood. The present study was developed in the Central Depression of Rio Grande do Sul and aimed to determine variations in pollen viability along flowering, ability to perform autogamy and dye efficiency for the viability test. Pollen viability was determined by colorimetry as well as the ability to perform autogamy by isolating floral buds, being evaluated in eleven matrices. Average pollen viability was 81.43%, with the highest average value obtained with the dye 2,3,5- Triphenyltetrazolium Chloride (TTC (84.11%. Safranin was not a good indicator at the tested concentration. No correlation was found between pollen viability and flowering time. The species demonstrated an inability to perform autogamy.

  11. Economic viability of geriatric hip fracture centers.

    Science.gov (United States)

    Clement, R Carter; Ahn, Jaimo; Mehta, Samir; Bernstein, Joseph

    2013-12-01

    Management of geriatric hip fractures in a protocol-driven center can improve outcomes and reduce costs. Nonetheless, this approach has not spread as broadly as the effectiveness data would imply. One possible explanation is that operating such a center is not perceived as financially worthwhile. To assess the economic viability of dedicated hip fracture centers, the authors built a financial model to estimate profit as a function of costs, reimbursement, and patient volume in 3 settings: an average US hip fracture program, a highly efficient center, and an academic hospital without a specific hip fracture program. Results were tested with sensitivity analysis. A local market analysis was conducted to assess the feasibility of supporting profitable hip fracture centers. The results demonstrate that hip fracture treatment only becomes profitable when the annual caseload exceeds approximately 72, assuming costs characteristic of a typical US hip fracture program. The threshold of profitability is 49 cases per year for high-efficiency hip fracture centers and 151 for the urban academic hospital under review. The largest determinant of profit is reimbursement, followed by costs and volume. In the authors’ home market, 168 hospitals offer hip fracture care, yet 85% fall below the 72-case threshold. Hip fracture centers can be highly profitable through low costs and, especially, high revenues. However, most hospitals likely lose money by offering hip fracture care due to inadequate volume. Thus, both large and small facilities would benefit financially from the consolidation of hip fracture care at dedicated hip fracture centers. Typical US cities have adequate volume to support several such centers.

  12. Is cell viability always directly related to corrosion resistance of stainless steels?

    International Nuclear Information System (INIS)

    Salahinejad, E.; Ghaffari, M.; Vashaee, D.; Tayebi, L.

    2016-01-01

    It has been frequently reported that cell viability on stainless steels is improved by increasing their corrosion resistance. The question that arises is whether human cell viability is always directly related to corrosion resistance in these biostable alloys. In this work, the microstructure and in vitro corrosion behavior of a new class of medical-grade stainless steels were correlated with adult human mesenchymal stem cell viability. The samples were produced by a powder metallurgy route, consisting of mechanical alloying and liquid-phase sintering with a sintering aid of a eutectic Mn–Si alloy at 1050 °C for 30 and 60 min, leading to nanostructures. In accordance with transmission electron microscopic studies, the additive particles for the sintering time of 30 min were not completely melted. Electrochemical impedance spectroscopic experiments suggested the higher corrosion resistance for the sample sintered for 60 min; however, a better cell viability on the surface of the less corrosion-resistant sample was unexpectedly found. This behavior is explained by considering the higher ion release rate of the Mn–Si additive material, as preferred sites to corrosion attack based on scanning electron microscopic observations, which is advantageous to the cells in vitro. In conclusion, cell viability is not always directly related to corrosion resistance in stainless steels. Typically, the introduction of biodegradable and biocompatible phases to biostable alloys, which are conventionally anticipated to be corrosion-resistant, can be advantageous to human cell responses similar to biodegradable metals. - Highlights: • Cell viability vs. corrosion resistance for medical-grade stainless steels • The stainless steel samples were prepared by powder metallurgy. • Unpenetrated additive played a critical role in the correlation.

  13. Is cell viability always directly related to corrosion resistance of stainless steels?

    Energy Technology Data Exchange (ETDEWEB)

    Salahinejad, E., E-mail: salahinejad@kntu.ac.ir [Faculty of Materials Science and Engineering, K.N. Toosi University of Technology, Tehran (Iran, Islamic Republic of); Ghaffari, M. [Bruker AXS Inc., 5465 East Cheryl Parkway, Madison, WI 53711 (United States); Vashaee, D. [Electrical and Computer Engineering Department, North Carolina State University, Raleigh, NC 27606 (United States); Tayebi, L. [Department of Developmental Sciences, Marquette University School of Dentistry, Milwaukee, WI 53201 (United States); Department of Engineering Science, University of Oxford, Oxford OX1 3PJ (United Kingdom)

    2016-05-01

    It has been frequently reported that cell viability on stainless steels is improved by increasing their corrosion resistance. The question that arises is whether human cell viability is always directly related to corrosion resistance in these biostable alloys. In this work, the microstructure and in vitro corrosion behavior of a new class of medical-grade stainless steels were correlated with adult human mesenchymal stem cell viability. The samples were produced by a powder metallurgy route, consisting of mechanical alloying and liquid-phase sintering with a sintering aid of a eutectic Mn–Si alloy at 1050 °C for 30 and 60 min, leading to nanostructures. In accordance with transmission electron microscopic studies, the additive particles for the sintering time of 30 min were not completely melted. Electrochemical impedance spectroscopic experiments suggested the higher corrosion resistance for the sample sintered for 60 min; however, a better cell viability on the surface of the less corrosion-resistant sample was unexpectedly found. This behavior is explained by considering the higher ion release rate of the Mn–Si additive material, as preferred sites to corrosion attack based on scanning electron microscopic observations, which is advantageous to the cells in vitro. In conclusion, cell viability is not always directly related to corrosion resistance in stainless steels. Typically, the introduction of biodegradable and biocompatible phases to biostable alloys, which are conventionally anticipated to be corrosion-resistant, can be advantageous to human cell responses similar to biodegradable metals. - Highlights: • Cell viability vs. corrosion resistance for medical-grade stainless steels • The stainless steel samples were prepared by powder metallurgy. • Unpenetrated additive played a critical role in the correlation.

  14. Link overlap, viability, and mutual percolation in multiplex networks

    International Nuclear Information System (INIS)

    Min, Byungjoon; Lee, Sangchul; Lee, Kyu-Min; Goh, K.-I.

    2015-01-01

    Many real-world complex systems are best modeled by multiplex networks. The multiplexity has proved to have broad impact on the system’s structure and function. Most theoretical studies on multiplex networks to date, however, have largely ignored the effect of the link overlap across layers despite strong empirical evidences for its significance. In this article, we investigate the effect of the link overlap in the viability of multiplex networks, both analytically and numerically. After a short recap of the original multiplex viability study, the distinctive role of overlapping links in viability and mutual connectivity is emphasized and exploited for setting up a proper analytic framework. A rich phase diagram for viability is obtained and greatly diversified patterns of hysteretic behavior in viability are observed in the presence of link overlap. Mutual percolation with link overlap is revisited as a limit of multiplex viability problem, and the controversy between existing results is clarified. The distinctive role of overlapping links is further demonstrated by the different responses of networks under random removals of overlapping and non-overlapping links, respectively, as well as under several link-removal strategies. Our results show that the link overlap facilitates the viability and mutual percolation; at the same time, the presence of link overlap poses a challenge in analytical approaches to the problem

  15. Viability and Virulence of Entomopathogenic Nematodes Exposed to Ultraviolet Radiation.

    Science.gov (United States)

    Shapiro-Ilan, David I; Hazir, Selcuk; Lete, Luis

    2015-09-01

    Entomopathogenic nematodes (EPNs) can be highly effective biocontrol agents, but their efficacy can be reduced due to exposure to environmental stress such as from ultraviolet (UV) radiation. Our objectives were to 1) compare UV tolerance among a broad array of EPN species, and 2) investigate the relationship between reduced nematode viability (after exposure to UV) and virulence. Nematodes exposed to a UV radiation (254 nm) for 10 or 20 min were assessed separately for viability (survival) and virulence to Galleria mellonella. We compared 9 different EPN species and 15 strains: Heterorhabditis bacteriophora (Baine, fl11, Oswego, and Vs strains), H. floridensis (332), H. georgiana (Kesha), H. indica (HOM1), H. megidis (UK211), Steinernema carpocapsae (All, Cxrd, DD136, and Sal strains), S. feltiae (SN), S. rarum (17C&E), and S. riobrave (355). In viability assessments, steinernematids, particularly strains of S. carpocapsae, generally exhibited superior UV tolerance compared with the heterorhabditids. However, some heterorhabditids tended to be more tolerant than others, e.g., H. megidis and H. bacteriophora (Baine) were most susceptible and H. bacteriophora (Vs) was the only heterorhabditid that did not exhibit a significant effect after 10 min of exposure. All heterorhabditids experienced reduced viability after 20 min exposure though several S. carpocapsae strains did not. In total, after 10 or 20 min exposure, the viability of seven nematode strains did not differ from their non-UV exposed controls. In virulence assays, steinernematids (particularly S. carpocapsae strains) also tended to exhibit higher UV tolerance. However, in contrast to the viability measurements, all nematodes experienced a reduction in virulence relative to their controls. Correlation analysis revealed that viability among nematode strains is not necessarily related to virulence. In conclusion, our results indicate that the impact of UV varies substantially among EPNs, and viability alone

  16. The differential effects of heat-shocking on the viability of spores from Bacillus anthracis, Bacillus subtilis, and Clostridium sporogenes after treatment with peracetic acid- and glutaraldehyde-based disinfectants.

    Science.gov (United States)

    March, Jordon K; Pratt, Michael D; Lowe, Chinn-Woan; Cohen, Marissa N; Satterfield, Benjamin A; Schaalje, Bruce; O'Neill, Kim L; Robison, Richard A

    2015-10-01

    This study investigated (1) the susceptibility of Bacillus anthracis (Ames strain), Bacillus subtilis (ATCC 19659), and Clostridium sporogenes (ATCC 3584) spores to commercially available peracetic acid (PAA)- and glutaraldehyde (GA)-based disinfectants, (2) the effects that heat-shocking spores after treatment with these disinfectants has on spore recovery, and (3) the timing of heat-shocking after disinfectant treatment that promotes the optimal recovery of spores deposited on carriers. Suspension tests were used to obtain inactivation kinetics for the disinfectants against three spore types. The effects of heat-shocking spores after disinfectant treatment were also determined. Generalized linear mixed models were used to estimate 6-log reduction times for each spore type, disinfectant, and heat treatment combination. Reduction times were compared statistically using the delta method. Carrier tests were performed according to AOAC Official Method 966.04 and a modified version that employed immediate heat-shocking after disinfectant treatment. Carrier test results were analyzed using Fisher's exact test. PAA-based disinfectants had significantly shorter 6-log reduction times than the GA-based disinfectant. Heat-shocking B. anthracis spores after PAA treatment resulted in significantly shorter 6-log reduction times. Conversely, heat-shocking B. subtilis spores after PAA treatment resulted in significantly longer 6-log reduction times. Significant interactions were also observed between spore type, disinfectant, and heat treatment combinations. Immediately heat-shocking spore carriers after disinfectant treatment produced greater spore recovery. Sporicidal activities of disinfectants were not consistent across spore species. The effects of heat-shocking spores after disinfectant treatment were dependent on both disinfectant and spore species. Caution must be used when extrapolating sporicidal data of disinfectants from one spore species to another. Heat

  17. R/L, a double reporter mouse line that expresses luciferase gene upon Cre-mediated excision, followed by inactivation of mRFP expression.

    Science.gov (United States)

    Jia, Junshuang; Lin, Xiaolin; Lin, Xia; Lin, Taoyan; Chen, Bangzhu; Hao, Weichao; Cheng, Yushuang; Liu, Yu; Dian, Meijuan; Yao, Kaitai; Xiao, Dong; Gu, Weiwang

    2016-10-01

    The Cre/loxP system has become an important tool for the conditional gene knockout and conditional gene expression in genetically engineered mice. The applications of this system depend on transgenic reporter mouse lines that provide Cre recombinase activity with a defined cell type-, tissue-, or developmental stage-specificity. To develop a sensitive assay for monitoring Cre-mediated DNA excisions in mice, we generated Cre-mediated excision reporter mice, designated R/L mice (R/L: mRFP(monomeric red fluorescent protein)/luciferase), express mRFP throughout embryonic development and adult stages, while Cre-mediated excision deletes a loxP-flanked mRFP reporter gene and STOP sequence, thereby activating the expression of the second reporter gene luciferase, as assayed by in vivo and ex vivo bioluminescence imaging. After germ line deletion of the floxed mRFP and STOP sequence in R/L mice by EIIa-Cre mice, the resulting luciferase transgenic mice in which the loxP-mRFP-STOP-loxP cassette is excised from all cells express luciferase in all tissues and organs examined. The expression of luciferase transgene was activated in liver of RL/Alb-Cre double transgenic mice and in brain of RL/Nestin-Cre double transgenic mice when R/L reporter mice were mated with Alb-Cre mice and Nestin-Cre mice, respectively. Our findings reveal that the double reporter R/L mouse line is able to indicate the occurrence of Cre-mediated excision from early embryonic to adult lineages. Taken together, these findings demonstrate that the R/L mice serve as a sensitive reporter for Cre-mediated DNA excision both in living animals and in organs, tissues, and cells following necropsy.

  18. Gamma camera based Positron Emission Tomography: a study of the viability on quantification; Tomografia por emissao de positrons com sistemas PET/SPECT: um estudo da viabilidade de quantifizacao

    Energy Technology Data Exchange (ETDEWEB)

    Pozzo, Lorena

    2005-07-01

    Positron Emission Tomography (PET) is a Nuclear Medicine imaging modality for diagnostic purposes. Pharmaceuticals labeled with positron emitters are used and images which represent the in vivo biochemical process within tissues can be obtained. The positron/electron annihilation photons are detected in coincidence and this information is used for object reconstruction. Presently, there are two types of systems available for this imaging modality: the dedicated systems and those based on gamma camera technology. In this work, we utilized PET/SPECT systems, which also allows for the traditional Nuclear Medicine studies based on single photon emitters. There are inherent difficulties which affect quantification of activity and other indices. They are related to the Poisson nature of radioactivity, to radiation interactions with patient body and detector, noise due to statistical nature of these interactions and to all the detection processes, as well as the patient acquisition protocols. Corrections are described in the literature and not all of them are implemented by the manufacturers: scatter, attenuation, random, decay, dead time, spatial resolution, and others related to the properties of each equipment. The goal of this work was to assess these methods adopted by two manufacturers, as well as the influence of some technical characteristics of PET/SPECT systems on the estimation of SUV. Data from a set of phantoms were collected in 3D mode by one camera and 2D, by the other. We concluded that quantification is viable in PET/SPECT systems, including the estimation of SUVs. This is only possible if, apart from the above mentioned corrections, the camera is well tuned and coefficients for sensitivity normalization and partial volume corrections are applied. We also verified that the shapes of the sources used for obtaining these factors play a role on the final results and should be delt with carefully in clinical quantification. Finally, the choice of the region

  19. Viability of and Escherichia coli O157:H7 and Listeria monocytogenes in a delicatessen appetizer (yogurt-based) salad as affected by citrus extract (Citrox©) and storage temperature.

    Science.gov (United States)

    Tsiraki, Maria I; Yehia, Hany M; Elobeid, Tahra; Osaili, Tareq; Sakkas, Hercules; Savvaidis, Ioannis N

    2018-02-01

    The antimicrobial effect of citrus extract (at 1 mL/kg [C1] and 2 mL/kg [C2]) on naturally occurring microbiota and inoculated pathogens (E. coli O157:H7 and L. monocytogenes at ca. 6 log cfu/g) in the traditional Greek yogurt-based salad Tzatziki stored at 4, 10, or 21 °C, was examined. Lactic acid bacteria (LAB) were high (8.0-8.5 log cfu/g) and varied only minimally for both the control (untreated) and the citrus extract-treated salad samples, whereas the higher citrus extract concentration yielded the lowest yeast populations, irrespective of temperature, during the entire storage period. Populations of inoculated E. coli (6 log cfu/g) declined in both untreated and citrus extract-treated samples from day 0-70, 35, and 15 at 4, 10, and 21 °C, respectively. Citrus extract had a significant effect on the survival of the inoculated E. coli O157:H7, with reductions of 2.8-4.8 log cfu/g in the citrus extract-treated samples at the end of the storage period. Our data show that L. monocytogenes survived in both untreated and citrus extract-treated samples during the entire storage period, irrespective of the storage temperature. The higher concentration of citrus extract had a significant effect on the survival of L. monocytogenes in the treated samples, and reductions of 1.5-3.0 logs were noted on final day 70, 35 and 15 at 4, 10 and 21 °C, respectively. The results of our study demonstrated the potential of citrus extract as a natural compound that can control the growth of food-borne pathogenic bacteria, such as E. coli O157:H7 and L. monocytogenes in Tzatziki, a yogurt-based salad. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Scaffold Architecture Controls Insulinoma Clustering, Viability, and Insulin Production

    Science.gov (United States)

    Blackstone, Britani N.; Palmer, Andre F.; Rilo, Horacio R.

    2014-01-01

    Recently, in vitro diagnostic tools have shifted focus toward personalized medicine by incorporating patient cells into traditional test beds. These cell-based platforms commonly utilize two-dimensional substrates that lack the ability to support three-dimensional cell structures seen in vivo. As monolayer cell cultures have previously been shown to function differently than cells in vivo, the results of such in vitro tests may not accurately reflect cell response in vivo. It is therefore of interest to determine the relationships between substrate architecture, cell structure, and cell function in 3D cell-based platforms. To investigate the effect of substrate architecture on insulinoma organization and function, insulinomas were seeded onto 2D gelatin substrates and 3D fibrous gelatin scaffolds with three distinct fiber diameters and fiber densities. Cell viability and clustering was assessed at culture days 3, 5, and 7 with baseline insulin secretion and glucose-stimulated insulin production measured at day 7. Small, closely spaced gelatin fibers promoted the formation of large, rounded insulinoma clusters, whereas monolayer organization and large fibers prevented cell clustering and reduced glucose-stimulated insulin production. Taken together, these data show that scaffold properties can be used to control the organization and function of insulin-producing cells and may be useful as a 3D test bed for diabetes drug development. PMID:24410263

  1. Viability of bull semen extended with commercial semen extender ...

    African Journals Online (AJOL)

    Andrea Raseona

    stored at 24 °C. Sperm motility parameters, morphology, and viability were analysed ... body size, slow average daily weight gain, decreased fertility, extended .... were determined by counting a total of 200 spermatozoa per each stained slide.

  2. Evaluation of pollen viability, stigma receptivity and fertilization ...

    African Journals Online (AJOL)

    AJL

    2013-11-13

    Nov 13, 2013 ... by field artificial pollination were analyzed in this study. The maximum pollen viability .... the day before anthesis to avoid self-pollination. Subsequently, between ..... The Lagerstroemia handbook/checklist. Ameri. Association ...

  3. Influence of gamma irradiation on pollen viability, germination ability ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-12-15

    Dec 15, 2009 ... minor cross-incompatibilities and physiological studies of incompatibility ... campanula shape of the flowers attractive for insect. (bees are ..... irradiation of cacao (Theobroma cacao L.) pollen: Effect on pollen grain viability ...

  4. In vitro pollen quantity, viability and germination tests in quince

    African Journals Online (AJOL)

    use

    2011-11-21

    Nov 21, 2011 ... rootstock pollens were collected in April from the unopened pink balloon-stage flowers on mature trees ..... their pollen characteristics to be used in cross breeding ... viability and germination levels for pistachio and terebinth in.

  5. Equine ovarian tissue viability after cryopreservation and in vitro culture

    Science.gov (United States)

    The efficiency of several cryoprotective agents were compared using both slow-freezing and vitrification methods. Results indicate that the viability of ovarian tissue cells increases when DMSO (slow-freezing) and ethylene glycol (vitrification) are used....

  6. Approximate viability for nonlinear evolution inclusions with application to controllability

    Directory of Open Access Journals (Sweden)

    Omar Benniche

    2016-12-01

    Full Text Available We investigate approximate viability for a graph with respect to fully nonlinear quasi-autonomous evolution inclusions. As application, an approximate null controllability result is given.

  7. Puget Sound steelhead life cycle model analyses - Population Viability Analysis

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This research was initiated by the Puget Sound Steelhead Technical Recovery Team to develop viability criteria for threatened Puget Sound steelhead and to support...

  8. STUDY ON POLLEN VIABILITY AS BIOINDICATOR OF AIR QUALITY

    Directory of Open Access Journals (Sweden)

    Florentina ŞTEFLEA

    2012-01-01

    Full Text Available The aim of this research is to estimate the relationship between pollen viability and atmospheric pollution (in polluted and non-polluted conditions. The study was carried out in the city of Timisoara. Two areas, with different intensity of road traffic (very high and absent but all characterized by the presence of the same plant species, were selected. The pollen of herbaceous spontaneous species, arboreal species and a shrub species was used (Robinia pseudacacia, Aesculus x carnea, Catalpa bignonioides, Albizzia julibrissin, Rosa canina, Sambucus nigra, Malva neglecta, Ranunculus acer, Trifolium repens, Cichorium intybus. The pollen of these species was treated with TTC (2, 3, 5 Tryphenil-Tetrazolium-Chloride staining solution and viability was then estimated by light microscopy. The results of the mean pollen viability percentage of the examined species are reported. Pollen viability of herbaceous plants is significantly different between the two environments.

  9. Pollen diversity, viability and floral structure of some Musa genotypes

    African Journals Online (AJOL)

    Prof. Ogunji

    Pollen diversity, viability and floral structure of some Musa genotypes ... at the Faculty of Agriculture & Natural Resources Management farm, Ebonyi State University,. Abakaliki. ..... Roots, tuber, plantains and bananas in human nutrition. Rome,.

  10. Viabilidade da estimativa de QI a partir dos subtestes Vocabulário e Cubos da WAIS-III Viability of IQ estimates based on Vocabulary and Block Design WAIS-III subtests

    Directory of Open Access Journals (Sweden)

    Flávia Wagner

    2010-08-01

    Full Text Available Objetivou-se avaliar a viabilidade de estimar o Quociente de Inteligência (QI de adultos por meio de duas estratégias que utilizam uma forma curta constituída pelos subtestes Vocabulário e Cubos da Escala Wechsler de Inteligência para Adultos (WAIS-III. Participaram do estudo 77 homens e mulheres, com idade média de 39,1 anos (dp=13,4. Foi aplicada a WAIS-III e questionários para cumprir os critérios de inclusão dos participantes sem prejuízos neurológicos e psiquiátricos. Para avaliar a concordância entre as duas estratégias de estimar o QI, utilizou-se o método estatístico proposto por Bland e Altman. Como resultado, encontrou-se que as estimativas de QI geradas pelas duas estratégias apresentam concordância com a medida do QI realizada pela aplicação da escala completa apenas em alguns dos casos avaliados. Sugere-se cautela na utilização dessas estratégias para estimar o QI de adultos.The aim of this study was to evaluate the feasibility of estimating the Intelligence Quotient (IQ in adults based on two strategies that use Vocabulary and Block Design subtests of Wechsler Adult Intelligence Scale (WAIS-III. Seventy-seven men and women participated of the study, with mean age of 39.1 (sd=13.4. WAIS-III subtests were administered and questionnaires were used to verify inclusion criteria for participants without neurological or psychiatric impairments. A statistical method proposed by Bland and Altman was used to evaluate the agreement between both strategies to estimate IQ. Results suggest that IQ estimates derived from both strategies only agree with the IQ derived from the full battery in a few cases of the sample. Considering these findings, caution is suggested when using these strategies to estimate adults' IQ.

  11. Demographic rates and population viability of black bears in Louisiana

    Science.gov (United States)

    Laufenberg, Jared S.; Clark, Joseph D.; Hooker, Michael J.; Lowe, Carrie L.; O'Connell-Goode, Kaitlin C.; Troxler, Jesse C.; Davidson, Maria M.; Chamberlain, Michael J.; Chandler, Richard B.

    2015-01-01

    observed 43 yearling litters for 33 females in the TRB and 21 yearling litters for 19 females in the TRC. The estimated number of cubs and number of yearlings produced per breeding adult female was 0.47 and 0.20, respectively, in the TRB and 0.32 and 0.18 in the TRC. On the basis of matrix projection models, asymptotic growth rates ranged from 1.053 to 1.078 for the TRB and from 1.005 to 1.062 for the TRC, depending on how we treated unresolved fates of adult females. Persistence probabilities estimated from stochastic population models based on telemetry data ranged from 0.997 to 0.998 for the TRC subpopulation depending on model assumptions and were >0.999 for the TRB regardless of model assumptions. We extracted DNA from hair collected at baited, barbed-wire enclosures in the TRB, UARB, and LARB to determine individual identities for capture-mark-recapture (CMR) analysis. We used those detection histories to estimate apparent survival (φ), per-capita recruitment (f), abundance (N), realized growth rate (λ), and long-term viability, based on Bayesian hierarchical modeling methods that allowed estimation of temporal process variance and parameter uncertainty. Based on 23,312 hair samples, annual N for females in the TRB ranged from 133 to 164 during 2006–2012, depending on year and how detection heterogeneity was modeled. Geometric mean of λ ranged from 0.996 to 1.002. In the UARB, we collected 11,643 hair samples from 2007 to 2012, from which estimates of N for females ranged from 23 to 43 during the study period, depending on detection heterogeneity model. The geometric mean of λ ranged from 1.038 to 1.059. Estimated N for females in LARB ranged from 69 to 96, and annual λ ranged from 0.80 to 1.11 based on 3,698 hair samples collected during 2010–2012, also depending on year and heterogeneity model. Probabilities of persistence over 100 years for the TRC and TRB based on stochastic matrix projection models that used vital rate estimates from telemetry data were

  12. Stand-Alone Solar Organic Rankine Cycle Water Pumping System and Its Economic Viability in Nepal

    OpenAIRE

    Suresh Baral; Kyung Chun Kim

    2015-01-01

    The current study presents the concept of a stand-alone solar organic Rankine cycle (ORC) water pumping system for rural Nepalese areas. Experimental results for this technology are presented based on a prototype. The economic viability of the system was assessed based on solar radiation data of different Nepalese geographic locations. The mechanical power produced by the solar ORC is coupled with a water pumping system for various applications, such as drinking and irrigation. The thermal ef...

  13. Enhancement of sperm motility and viability by turmeric by-product dietary supplementation in roosters.

    Science.gov (United States)

    Yan, Wenjing; Kanno, Chihiro; Oshima, Eiki; Kuzuma, Yukiko; Kim, Sung Woo; Bai, Hanako; Takahashi, Masashi; Yanagawa, Yojiro; Nagano, Masashi; Wakamatsu, Jun-Ichi; Kawahara, Manabu

    2017-10-01

    Improving sperm motility and viability are major goals to improve efficiency in the poultry industry. In this study, the effects of supplemental dietary turmeric by-product (TBP) from commercial turmeric production on sperm motility, viability, and antioxidative status were examined in domestic fowl. Mature Rhode Island Red roosters were divided into two groups - controls (groupC) without TBP administration and test subjects (groupT) fed a basal diet supplemented with 0.8g of TBP/day in a temperature-controlled rearing facility (Experiment 1) and 1.6g/day under heat stress (Experiment 2) for 4 weeks. In Experiment 1, TBP dietary supplementation increased the sperm motility variables straight-line velocity, curvilinear velocity, and linearity based on a computer-assisted semen analysis, 2 weeks following TBP supplementation. In Experiment 2, using flow cytometry, sperm viability at 3 and 4 weeks following TBP supplementation was greater in Group T than C, and this increase was consistent with a reduction in reactive oxygen species (ROS) production at 2 and 4 weeks. The results of both experiments clearly demonstrate that dietary supplementation with TBP enhanced sperm motility in the controlled-temperature conditions as well as sperm viability, and reduced ROS generation when heat stress prevailed. Considering its potential application in a range of environments, TBP may serve as an economical and potent antioxidant to improve rooster fertility. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Viability and Biochemical Content Changes in Seed Storage of Jabon Putih (Anthocephalus Cadamba (Roxb Miq.

    Directory of Open Access Journals (Sweden)

    Naning Yuniarti

    2015-08-01

    Full Text Available Seed deterioration is the process of deteriorated seed in view of viability that changed the physiological and chemical content. The aim of this research is to find out the effect of moisture content and storage on viability and biochemical content of jabon putih seed. Completely Randomized Design was used for decreasing moisture content based on seed drying time for 0, 24, 48, 72, 96, 120 hours, and Completely Randomized Factorial Design was used for the combination of moisture content (drying time for 0, 24, 48, 72, 96, 120 hours and room storage (ambient room, air-conditioned room, refrigerator. The results of this research are: (1 seed drying time and seed storage affected the change of seed viability and biochemical content, (2 seed drying time and seed storage influence significantly the value of moisture content, germination percentage, and biochemical content (lipid, carbohydrate, protein, (3 The longer time of seed drying and seed storage has decreased the seed moisture content, germination percentage, and the carbohydrate content, but it has increased the content of lipid and protein, and (4 the seeds were stored in refrigerator has better viability compared to dry cold storage and ambient room.

  15. Solubilization and folding of a fully active recombinant Gaussia luciferase with native disulfide bonds by using a SEP-Tag.

    Science.gov (United States)

    Rathnayaka, Tharangani; Tawa, Minako; Nakamura, Takashi; Sohya, Shihori; Kuwajima, Kunihiro; Yohda, Masafumi; Kuroda, Yutaka

    2011-12-01

    Gaussia luciferase (GLuc) is the smallest known bioluminescent protein and is attracting much attention as a potential reporter protein. However, its 10 disulfide bond forming cysteines have hampered the efficient production of recombinant GLuc and thus limited its use in bio-imaging application. Here, we demonstrate that the addition of a short solubility enhancement peptide tag (SEP-Tag) to the C-terminus of GLuc (GLuc-C9D) significantly increased the fraction of soluble protein at a standard expression temperature. The expression time was much shorter, and the final yield of GLuc-C9D was significantly higher than with our previous pCold expression system. Reversed phase HPLC indicated that the GLuc-C9D variant folded with a single disulfide bond pattern after proper oxidization. Further, the thermal denaturation of GLuc-C9D was completely reversible, and its secondary structure content remained unchanged until 40°C as assessed by CD spectroscopy. The (1)H-NMR spectrum of GLuc indicated sharp well dispersed peaks typical for natively folded proteins. GLuc-C9D bioluminescence activity was strong and fully retained even after incubation at high temperatures. These results suggest that solubilization using SEP-Tags can be useful for producing large quantities of proteins containing multiple disulfide bonds. Copyright © 2011. Published by Elsevier B.V.

  16. High-throughput screening of effective siRNAs using luciferase-linked chimeric mRNA.

    Directory of Open Access Journals (Sweden)

    Shen Pang

    Full Text Available The use of siRNAs to knock down gene expression can potentially be an approach to treat various diseases. To avoid siRNA toxicity the less transcriptionally active H1 pol III promoter, rather than the U6 promoter, was proposed for siRNA expression. To identify highly efficacious siRNA sequences, extensive screening is required, since current computer programs may not render ideal results. Here, we used CCR5 gene silencing as a model to investigate a rapid and efficient screening approach. We constructed a chimeric luciferase-CCR5 gene for high-throughput screening of siRNA libraries. After screening approximately 900 shRNA clones, 12 siRNA sequences were identified. Sequence analysis demonstrated that most (11 of the 12 sequences of these siRNAs did not match those identified by available siRNA prediction algorithms. Significant inhibition of CCR5 in a T-lymphocyte cell line and primary T cells by these identified siRNAs was confirmed using the siRNA lentiviral vectors to infect these cells. The inhibition of CCR5 expression significantly protected cells from R5 HIV-1JRCSF infection. These results indicated that the high-throughput screening method allows efficient identification of siRNA sequences to inhibit the target genes at low levels of expression.

  17. Experimental evidence for the physiological role of bacterial luciferase in the protection of cells against oxidative stress.

    Science.gov (United States)

    Szpilewska, Hanna; Czyz, Agata; Wegrzyn, Grzegorz

    2003-11-01

    The origin and function of bioluminescence was considered a problematic question of the Charles Darwin theory. Early evolution of bacterial luminescence and its current physiological importance seem to be especially mysterious. Recently, it was proposed that stimulation of DNA repair may be a physiological role for production of light by bacterial cells. On the other hand, it was also proposed that primary role of luminescent systems could be detoxification of the deleterious oxygen derivatives. Although some previous results might suggest that this hypothesis can be correct, until now experimental evidence for such a mechanism operating in bacterial cells and having physiological importance was generally lacking. Here we demonstrate that in the presence of various oxidants (hydrogen peroxide, cumene hydroperoxide, t-butyl hydroperoxide, and ferrous ions) at certain concentrations in the culture medium, growth of Vibrio harveyi mutants luxA and luxB, but not of the mutant luxD, is severely impaired relative to wild-type bacteria. This deleterious effect of oxidants on the mutants luxA and luxB could be significantly reduced by addition of the antioxidants A-TEMPO or 40H-TEMPO. We conclude that bacterial luciferase may indeed play a physiological role in the protection of cells against oxidative stress.

  18. Sonodelivery Facilitates Sustained Luciferase Expression from an Episomal Vector in Skeletal Muscle

    Directory of Open Access Journals (Sweden)

    Manoel Figueiredo Neto

    2015-07-01

    Full Text Available Successful gene delivery to skeletal muscle is a desirable goal, not only for treating muscle diseases, but also for immunization, treatment of metabolic disorders, and/or delivering gene expression that can treat systemic conditions, such as bone metastatic cancer, for example. Although naked DNA uptake into skeletal muscle is possible, it is largely inefficient in the absence of additional chemical or physical delivery methods. We describe a system for delivery of non-viral or plasmid DNA to skeletal muscle using ultrasound-assisted sonoporation of a nanoplex combining plasmid DNA and a branched polymer based on poly(cyclooctene-graft-oligopeptide. The materials and methods described herein promise to advance the field of sonodelivery and of gene delivery to muscle for therapeutic applications since a simple system is presented that enables long-term gene expression in vivo with the promise of a minimal inflammatory gene expression profile.

  19. Preservation of seed viability during 25 years of storage under standard genebank conditions

    NARCIS (Netherlands)

    Treuren, van R.; Groot, de E.C.; Hintum, van T.J.L.

    2013-01-01

    Maintaining sufficient viability is critical to the sustainability of ex situ conserved seed collections. For this reason, accessions are regenerated when viability falls below a predefined threshold. Viability is monitored by determining the germination ability of accessions at predefined time

  20. Determination of Lactic Acid Bacteria Viability in the Small Intestine of Catfish (Pangasius djambal by Using the 32P Radioisotope

    Directory of Open Access Journals (Sweden)

    I. Sugoro

    2015-04-01

    Full Text Available The viability of probiotics is important to be determined, as is its probiotic potency in the small instestine of fish. The result can be used as a basis to determine the feeding frequency of the probiotics to the fish.The aim of this study is to gain information about the viability of lactic acid bacteria (LAB in the small intestine of fish by using the 32P isotope technique. Catfish (Pangasius djambal was used as a test fish, and the LAB with the code of P2.1 PTB was the subject of the experiment. Before its viability was tested, the LAB had been labelled with radioisotope 32P, then mixed into catfish feed. Its viability could be determined by counting the activity of 32P. The results showed that the percentage of LAB viability in the small intestine of catfish declined until day 7. The percentage of LAB viability was decreased at an amount of 30% at day 3. Based on this result, the feeding frequency of LAB P2.1 PTB is every 3 days.

  1. Determination of Lactic Acid Bacteria Viability in the Small Intestine of Catfish (Pangasius djambal by Using the 32P Radioisotope

    Directory of Open Access Journals (Sweden)

    I. Sugoro

    2015-10-01

    Full Text Available The viability of probiotics is important to be determined, as is its probiotic potency in the small instestine of fish. The result can be used as a basis to determine the feeding frequency of the probiotics to the fish.The aim of this study is to gain information about the viability of lactic acid bacteria (LAB in the small intestine of fish by using the 32P isotope technique. Catfish (Pangasius djambal was used as a test fish, and the LAB with the code of P2.1 PTB was the subject of the experiment. Before its viability was tested, the LAB had been labelled with radioisotope 32P, then mixed into catfish feed. Its viability could be determined by counting the activity of 32P. The results showed that the percentage of LAB viability in the small intestine of catfish declined until day 7. The percentage of LAB viability was decreased at an amount of 30% at day 3. Based on this result, the feeding frequency of LAB P2.1 PTB is every 3 days. Received: 04 October 2014 Revised: 26 March 2015; Accepted: 05 April 2015

  2. Determination of lactic acid bacteria viability in the small intestine of catfish (Pangasius Djambal) by using the 32P radioisotope

    International Nuclear Information System (INIS)

    Sugoro, I.; Citraresmini, A.; Murni, A.P.; Fairuz, D.

    2015-01-01

    The viability of probiotics is important to be determined, as is its probiotics potency in the small intestine of fish. The result can be used as a basis to determine the feeding frequency of the probiotics to the fish. The aim of this study is to gain information about the viability of lactic acid bacteria (LAB) in the small intestine of fish by using the 32 P isotope technique. Catfish (Pangasius djambal) was used as a test fish, and the LAB with the code of P2.1 PTB was the subject of the experiment. Before its viability was tested, the LAB had been labelled with radioisotope 32 P, then mixed into catfish feed. Its viability could be determined by counting the activity of 32 P. The results showed that the percentage of LAB viability in the small intestine of catfish declined until day 7. The percentage of LAB viability was decreased at an amount of 30% at day 3. Based on this result, the feeding frequency of LAB P2.1 PTB is every 3 days. (author)

  3. Evaluating the Viability of Mobile Learning to Enhance Management Training

    Directory of Open Access Journals (Sweden)

    Iain Macdonald

    2011-04-01

    Full Text Available A qualitative research project was conducted to test the viability of augmenting an e-learning program for workplace learners using mobile content delivered through smart phones. Ten learners taking a six week web-based e-learning course were given smart phones which enabled them to access approximately 70% of the course content, in addition to having access to the full course via a standard e-learning website. Mobile content was provided in a variety of forms, including text, audio and video files, a mobile multiple-choice quiz website, and links to streaming videos. Study participants who were regular users of mobile phones found the mobile learning materials to be user-friendly, offering increased convenience and flexibility. Use of the mobile content tended to increase as learners spent more time in their day away from locations where Internet-linked computers could be found. Video was found to be the most effective means of presenting content, followed by audio and text. The most promising role of mobile learning appears to be to augment rather than replace e-learning or blended learning.

  4. Incorporating parametric uncertainty into population viability analysis models

    Science.gov (United States)

    McGowan, Conor P.; Runge, Michael C.; Larson, Michael A.

    2011-01-01

    Uncertainty in parameter estimates from sampling variation or expert judgment can introduce substantial uncertainty into ecological predictions based on those estimates. However, in standard population viability analyses, one of the most widely used tools for managing plant, fish and wildlife populations, parametric uncertainty is often ignored in or discarded from model projections. We present a method for explicitly incorporating this source of uncertainty into population models to fully account for risk in management and decision contexts. Our method involves a two-step simulation process where parametric uncertainty is incorporated into the replication loop of the model and temporal variance is incorporated into the loop for time steps in the model. Using the piping plover, a federally threatened shorebird in the USA and Canada, as an example, we compare abundance projections and extinction probabilities from simulations that exclude and include parametric uncertainty. Although final abundance was very low for all sets of simulations, estimated extinction risk was much greater for the simulation that incorporated parametric uncertainty in the replication loop. Decisions about species conservation (e.g., listing, delisting, and jeopardy) might differ greatly depending on the treatment of parametric uncertainty in population models.

  5. Viability of biocompatible and biodegradable seeds production with incorporated radionuclides

    International Nuclear Information System (INIS)

    Roberto, W.S.; Pereira, M.M.; Vasconcelos, W.L.; Campos, T.P.R.

    2000-01-01

    The present work aims the development of radioactive seeds, biocompatible and biodegradable, with the objective of adding options in the cancer treatment. The work focus on the production of seeds biodegradable that incorporate radioisotopes with half life inferior than the degradation time of the material. The idea of producing devices with biodegradable materials impregnated with radioisotopes of short half life will offer new possibilities in the cancer treatment, since they can be used following the same procedures of the permanent interstitial brachytherapy, but using degradable materials compatible with the physiological environment. It will be discussed in particular the possible application of these seeds in the treatment of prostate cancer. A review of the subject and a preliminary evaluation of the viability of production of the seeds will be presented. The method of production of the seeds is based on the incorporation of Iodine and Samarium in glass matrixes obtained by sol-gel processing. X-ray fluorescence was done in the samples produced and the incorporation of Iodine and Samarium atoms was confirmed. (author)

  6. Mobile dental operations: capital budgeting and long-term viability.

    Science.gov (United States)

    Arevalo, Oscar; Chattopadhyay, Amit; Lester, Harold; Skelton, Judy

    2010-01-01

    The University of Kentucky College of Dentistry (UKCD) runs a large mobile dental operation. Economic conditions dictate that as the mobile units age it will be harder to find donors willing or able to provide the financial resources for asset replacement. In order to maintain current levels of access for the underserved, consideration of replacement is paramount. A financial analysis for a new mobile unit was conducted to determine self-sustainability, return on investment (ROI), and feasibility of generating a cash reserve for its replacement in 12 years. Information on clinical income, operational and replacement costs, and capital costs was collected. A capital budgeting analysis (CBA) was conducted using the Net Present Value (NPV) methodology in four different scenarios. Depreciation funding was calculated by transferring funds from cash inflows and reinvested to offset depreciation at fixed compound interest. A positive ROI was obtained for two scenarios. He depreciation fund did not generate a cash reserve sufficient to replace the mobile unit. Mobile dental programs can play a vital role in providing access to care to underserved populations and ensuring their mission requires long-term planning. Careful financial viability and CBA based on sound assumptions are excellent decision-making tools.

  7. Nuclear hydrogen: An assessment of product flexibility and market viability

    International Nuclear Information System (INIS)

    Botterud, Audun; Yildiz, Bilge; Conzelmann, Guenter; Petri, Mark C.

    2008-01-01

    Nuclear energy has the potential to play an important role in the future energy system as a large-scale source of hydrogen without greenhouse gas emissions. Thus far, economic studies of nuclear hydrogen tend to focus on the levelized cost of hydrogen without accounting for the risks and uncertainties that potential investors would face. We present a financial model based on real options theory to assess the profitability of different nuclear hydrogen production technologies in evolving electricity and hydrogen markets. The model uses Monte Carlo simulations to represent uncertainty in future hydrogen and electricity prices. It computes the expected value and the distribution of discounted profits from nuclear hydrogen production plants. Moreover, the model quantifies the value of the option to switch between hydrogen and electricity production, depending on what is more profitable to sell. We use the model to analyze the market viability of four potential nuclear hydrogen technologies and conclude that flexibility in output product is likely to add significant economic value for an investor in nuclear hydrogen. This should be taken into account in the development phase of nuclear hydrogen technologies

  8. Assessment of Viability Appraisal Practice by Estate Surveyors and Valuers in Lagos Metropolis, Nigeria

    Directory of Open Access Journals (Sweden)

    Oyetunji Abiodun Kolawale

    2017-01-01

    Full Text Available A capital investment appraisal is a means of ensuring value for money. It is not meant to provide an indication of profit or loss, but rather a comparison of costs in relation to those areas where there is an opportunity or an inclination for change. The decision to invest in a project is based on the expectation of future returns since a rational investor does not expect to incur loss on his capital outlay. This decision making is usually hinged on advice obtained during pre-investment appraisal. This study is aimed at evaluating the mode of practicing investment viability appraisal by Estate Surveyors and Valuers in Lagos metropolis. The data for the study was obtained by randomly administering structured questionnaires on eighty-seven (87 practicing Estate Surveyors and Valuers in the study area and the findings were analyzed through descriptive statistical tools such as the Likert scale to present the result. Findings revealed that the payback period is the most adopted appraisal technique in practice as evidenced with a mean score of 3.57. The objective (s of the investor is also the most significant factor being considered while selecting appraisal technique with a mean score of 3.83 while the problems of actual return varying from the expected return and also difficulty in the repayment of loans always result when a wrong viability technique is employed. The success of any viability study goes beyond knowing the objective (s of the investor; therefore, it was recommended that appraisers should ensure that they are equipped with adequate knowledge required for the execution of feasibility and on viability studies because knowing the right viability criteria for a particular objective will help in advising an investor on a course of action that will best achieve the developer’s objective.

  9. Is cell viability always directly related to corrosion resistance of stainless steels?

    Science.gov (United States)

    Salahinejad, E; Ghaffari, M; Vashaee, D; Tayebi, L

    2016-05-01

    It has been frequently reported that cell viability on stainless steels is improved by increasing their corrosion resistance. The question that arises is whether human cell viability is always directly related to corrosion resistance in these biostable alloys. In this work, the microstructure and in vitro corrosion behavior of a new class of medical-grade stainless steels were correlated with adult human mesenchymal stem cell viability. The samples were produced by a powder metallurgy route, consisting of mechanical alloying and liquid-phase sintering with a sintering aid of a eutectic Mn-Si alloy at 1050 °C for 30 and 60 min, leading to nanostructures. In accordance with transmission electron microscopic studies, the additive particles for the sintering time of 30 min were not completely melted. Electrochemical impedance spectroscopic experiments suggested the higher corrosion resistance for the sample sintered for 60 min; however, a better cell viability on the surface of the less corrosion-resistant sample was unexpectedly found. This behavior is explained by considering the higher ion release rate of the Mn-Si additive material, as preferred sites to corrosion attack based on scanning electron microscopic observations, which is advantageous to the cells in vitro. In conclusion, cell viability is not always directly related to corrosion resistance in stainless steels. Typically, the introduction of biodegradable and biocompatible phases to biostable alloys, which are conventionally anticipated to be corrosion-resistant, can be advantageous to human cell responses similar to biodegradable metals. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Use of fluorescent redox indicators to evaluate cell proliferation and viability

    DEFF Research Database (Denmark)

    Rasmussen, E.S.

    1999-01-01

    The performance of two cell viability test kits based on the use of redox indicators yielding fluorescent products, the AlamarBlue assay and a resazurin-based in vitro toxicology assay kit from Sigma, was compared in the present study. Cultures of human neonatal foreskin fibroblasts were exposed...... for 168 h of continuous exposure, but showed equal levels of cytostatic effects in cultures with a low initial cell density after 72 h of exposure. Similar characteristics of the dye solutions were observed by high-performance Liquid chromatography (HPLC) separation and UV spectroscopy, and the major...... components were tentatively identified as resazurin and resorufin. The AlamarBlue assay has gained wide application as a cell viability indicator that allows continuous monitoring of cell proliferation or cytotoxicity in human and animal cells, bacteria, and fungi, but no studies with the deliberate use...

  11. Assessment of the viability of skin grafts

    International Nuclear Information System (INIS)

    Wahner, H.W.

    1988-01-01

    A number of tests are available to monitor the blood flow in free and distant pedicle skin grafts. The information from these tests aids in the development of measures to enhance vascularization and is occasionally needed to make clinical decisions in patients with distant pedicle grafts. Measurements of the disappearance of an intradermally injected small amount of 133 Xe allows determination of a clearance rate and blood flow before and after clamping the original blood supply through the base. With /sup 99m/Tc, which is generally more readily available, a flow index and block index can be determined. Clinically both procedures give equally good results in determining a safe time for pedicle base separation. The fluorescein test allows assessment of regional blood flow distribution within the pedicle

  12. A Luciferase-Expressing Leishmania braziliensis Line That Leads to Sustained Skin Lesions in BALB/c Mice and Allows Monitoring of Miltefosine Treatment Outcome.

    Directory of Open Access Journals (Sweden)

    Adriano C Coelho

    2016-05-01

    Full Text Available Leishmania braziliensis is the most prevalent species isolated from patients displaying cutaneous and muco-cutaneous leishmaniasis in South America. However, there are difficulties for studying L. braziliensis pathogenesis or response to chemotherapy in vivo due to the natural resistance of most mouse strains to infection with these parasites. The aim of this work was to develop an experimental set up that could be used to assess drug efficacy against L. braziliensis. The model was tested using miltefosine.A L. braziliensis line, originally isolated from a cutaneous leishmaniasis patient, was passaged repeatedly in laboratory rodents and further genetically manipulated to express luciferase. Once collected from a culture of parasites freshly transformed from amastigotes, 106 wild type or luciferase-expressing stationary phase promastigotes were inoculated subcutaneously in young BALB/c mice or golden hamsters. In both groups, sustained cutaneous lesions developed at the site of inoculation, no spontaneous self- healing being observed 4 months post-inoculation, if left untreated. Compared to the wild type line features, no difference was noted for the luciferase-transgenic line. Infected animals were treated with 5 or 15 mg/kg/day miltefosine orally for 15 days. At the end of treatment, lesions had regressed and parasites were not detected. However, relapses were observed in animals treated with both doses of miltefosine.Here we described experimental settings for a late-healing model of cutaneous leishmaniasis upon inoculation of a luciferase-expressing L. braziliensis line that can be applied to drug development projects. These settings allowed the monitoring of the transient efficacy of a short-term miltefosine administration.

  13. Drug Plant Seed Viability Preservation by Cryoconservation

    Directory of Open Access Journals (Sweden)

    Alexandra Sh. Dodonova

    2013-01-01

    Full Text Available The article considers the possibility of cryopreservation of seeds of several species of drug plants in Central Kazakhstan – Tanacetum ulutavicum, Niedzwedzkia semiretschenskia, Rhaponticum carthamoides. To increase the amount of viable seeds after liquid nitrogen freezing, we used different defrosting temperatures, deposited seeds with different moisture contents and used different containers for cryopreservation. Recommendations, concerning conditions of cryopreservation of seeds of these drug plant species were developed, basing on the obtained results.

  14. Warm Water Bath Stimulates Phase-Shifts of the Peripheral Circadian Clocks in PER2::LUCIFERASE Mouse

    Science.gov (United States)

    Kuriki, Daisuke; Haraguchi, Atsushi; Shibata, Shigenobu

    2014-01-01

    Circadian clocks in the peripheral tissues of mice are known to be entrained by pulse stimuli such as restricted feeding, novel wheel running, and several other agents. However, there are no reports on high temperature pulse-mediated entrainment on the phase-shift of peripheral clocks in vivo. Here we show that temperature treatment of mice for two days at 41°C, instead of 37°C, for 1–2 h during the inactive period, using a temperature controlled water bath stimulated phase-advance of peripheral clocks in the kidney, liver, and submandibular gland of PER2::LUCIFERASE mice. On the other hand, treatment for 2 days at 35°C ambient room temperature for 2 h did not cause a phase-advance. Maintenance of mice at 41°C in a water bath, sustained the core body temperature at 40–41°C. However, the use of 37°C water bath or the 35°C ambient room temperature elevated the core body temperature to 38.5°C, suggesting that at least a core body temperature of 40–41°C is necessary to cause phase-advance under light-dark cycle conditions. The temperature pulse stimulation at 41°C, instead of 37°C water bath for 2 h led to the elevated expression of Per1 and Hsp70 in the peripheral tissue of mice. In summary, the present study demonstrates that transient high temperature pulse using water bath during daytime causes phase-advance in mouse peripheral clocks in vivo. The present results suggest that hot water bath may affect the phase of peripheral clocks. PMID:24933288

  15. Aerosol-delivered programmed cell death 4 enhanced apoptosis, controlled cell cycle and suppressed AP-1 activity in the lungs of AP-1 luciferase reporter mice.

    Science.gov (United States)

    Hwang, S-K; Jin, H; Kwon, J T; Chang, S-H; Kim, T H; Cho, C-S; Lee, K H; Young, M R; Colburn, N H; Beck, G R; Yang, H-S; Cho, M-H

    2007-09-01

    The long-term survival of lung cancer patients treated with conventional therapies remains poor and therefore the need for novel approaches remains high. This has led to the re-emergence of aerosol delivery as a therapeutic intervention. In this study, glucosylated polyethylenimine (GPEI) was used as carrier to investigate programmed cell death 4 (PDCD4) and PDCD4 mutant (D418A), an eIF4A-binding mutant, on PDCD4-related signaling and activator protein-1 (AP-1) activity in the lungs of AP-1 luciferase reporter mice. After confirming the efficiency of GPEI as a carrier in lungs, the effects of aerosol-delivered PDCD4 were investigated in AP-1 luciferase reporter mice. Aerosol delivery of GPEI/PDCD4 through a nose-only inhalation facilitated the apoptosis of lungs whereas aerosol PDCD4 mutant did not. Also, such aerosol delivery regulated proteins relevant to cell-cycle control and suppressed AP-1 activity. Results obtained by western blot analysis, immunohistochemistry, luciferase assay and deoxynucleotidyl-transferase-mediated nick end labeling study suggest that combined actions such as facilitating apoptosis, controlling cell cycle and suppression of AP-1 activity by PDCD4 may provide useful tool for designing lung tumor prevention and treatment by which PDCD4 functions as a transformation suppressor in the future.

  16. Plasmodium falciparum transfected with ultra bright NanoLuc luciferase offers high sensitivity detection for the screening of growth and cellular trafficking inhibitors.

    Directory of Open Access Journals (Sweden)

    Mauro F Azevedo

    Full Text Available Drug discovery is a key part of malaria control and eradication strategies, and could benefit from sensitive and affordable assays to quantify parasite growth and to help identify the targets of potential anti-malarial compounds. Bioluminescence, achieved through expression of exogenous luciferases, is a powerful tool that has been applied in studies of several aspects of parasite biology and high throughput growth assays. We have expressed the new reporter NanoLuc (Nluc luciferase in Plasmodium falciparum and showed it is at least 100 times brighter than the commonly used firefly luciferase. Nluc brightness was explored as a means to achieve a growth assay with higher sensitivity and lower cost. In addition we attempted to develop other screening assays that may help interrogate libraries of inhibitory compounds for their mechanism of action. To this end parasites were engineered to express Nluc in the cytoplasm, the parasitophorous vacuole that surrounds the intraerythrocytic parasite or exported to the red blood cell cytosol. As proof-of-concept, these parasites were used to develop functional screening assays for quantifying the effects of Brefeldin A, an inhibitor of protein secretion, and Furosemide, an inhibitor of new permeation pathways used by parasites to acquire plasma nutrients.

  17. Viability tests of LIPI-MC mould collection in ampoule of L-drying preservation after one year of storage at 5ºC

    Directory of Open Access Journals (Sweden)

    MUHAMMAD ILYAS

    2007-01-01

    Full Text Available A study on viability test of several LIPI-MC mould isolates in ampoule of L-drying preservation after one year storage at 5º C had been conducted. In this study, cell survival level of 34 ampoules number from eight mould generas and 17 species had been counted. The objective of this study was to observe the survival or viability level of several mould isolates in ampoule L-drying preservation after one year storage at 5º C. The measurement of viability level was based on the average of colony forming unit density (CFU/ml. The result showed that there were seven mould isolates have hingh viabilitiy, 18 isolates have medium viabilitiy, two isolates have low viability, and five isolates have lost its viability.

  18. Multispectral imaging of organ viability during uterine transplantation surgery

    Science.gov (United States)

    Clancy, Neil T.; Saso, Srdjan; Stoyanov, Danail; Sauvage, Vincent; Corless, David J.; Boyd, Michael; Noakes, David E.; Thum, Meen-Yau; Ghaem-Maghami, Sadaf; Smith, J. R.; Elson, Daniel S.

    2014-02-01

    Uterine transplantation surgery has been proposed as a treatment for permanent absolute uterine factor infertility (AUFI) in the case of loss of the uterus. Due to the complexity of the vasculature correct reanastomosis of the blood supply during transplantation surgery is a crucial step to ensure reperfusion and viability of the organ. While techniques such as fluorescent dye imaging have been proposed to visualise perfusion there is no gold standard for intraoperative visualisation of tissue oxygenation. In this paper results from a liquid crystal tuneable filter (LCTF)-based multispectral imaging (MSI) laparoscope are described. The system was used to monitor uterine oxygen saturation (SaO2) before and after transplantation. Results from surgeries on two animal models (rabbits and sheep) are presented. A feature-based registration algorithm was used to correct for misalignment induced by breathing or peristalsis in the tissues of interest prior to analysis. An absorption spectrum was calculated at each spatial pixel location using reflectance data from a reference standard, and the relative contributions from oxy- and deoxyhaemoglobin were calculated using a least squares regression algorithm with non-negativity constraints. Results acquired during animal surgeries show that cornual oxygenation changes are consistent with those observed in point measurements taken using a pulse oximeter, showing reduced SaO2 following reanastomosis. Values obtained using the MSI laparoscope were lower than those taken with the pulse oximeter, which may be due to the latter's use of the pulsatile arterial blood signal. Future work incorporating immunological test results will help to correlate SaO2 levels with surgical outcomes.

  19. Methodological Aspects of Modeling Development and Viability of Systems and Counterparties in the Digital Economy

    Directory of Open Access Journals (Sweden)

    Vitlinskyy Valdemar V.

    2018-03-01

    Full Text Available The aim of the article is to study and generalize methodological approaches to modeling economic development and viability of economic systems with consideration for risk, changing their goals, status, and behavior in the digital economy. The definition of categories of economic development and viability is offered, the directions of their research by means of mathematical modeling are grounded. The system of characteristics and markers of the external economic environment under conditions of digitalization of economic activity is analyzed. The theoretical foundations and methodology for mathematical modeling of development of economic systems as well as ensuring their viability and security under conditions of introducing infrastructure of information society and digital economy on the principles of the information and knowledge approach are considered. It is proved that in an information society, predictive model technologies are a growing safety resource. There studied prerequisites for replacing the traditional integration concept of evaluation, analysis, modeling, management, and administration of economic development based on a threat-oriented approach to the definition of security protectors, information, and knowledge. There proposed a concept of creating a database of models for examining trends and patterns of economic development, which, unlike traditional trend models of dynamics, identifies and iteratively conceptualizes processes based on a set of knowledgeable predictors based on the use of data mining and machine learning tools, including in-depth training.

  20. Viability of seeds of two representatives from Apocynaceae family

    Directory of Open Access Journals (Sweden)

    Y.A. Aviekin

    2015-05-01

    Full Text Available The viability of some Apocynaceae seeds depending on the duration of storage under conditions of low temperature was studied. Extracted embryos from the seeds of Pachypodium lamerei Drake and Adenium obesum (Forssk. Roem. et Schult with different storage history were analyzed. Embryos were stained by acidic fuchsine what allows identification of viable and destructed cells. Destructed cells were stained much more intensively, while viable cells remained unstained. Observed results showed dependence of viability of P. lamerei and A. obesum seeds from term of storage. It was obtained that the seeds of investigated succulent species, just like in many other tropical plants, rapidly lost their viability and should be described as microbiotic.

  1. The Viability of Photocatalysis for Air Purification

    Directory of Open Access Journals (Sweden)

    Stephen O. Hay

    2015-01-01

    Full Text Available Photocatalytic oxidation (PCO air purification technology is reviewed based on the decades of research conducted by the United Technologies Research Center (UTRC and their external colleagues. UTRC conducted basic research on the reaction rates of various volatile organic compounds (VOCs. The knowledge gained allowed validation of 1D and 3D prototype reactor models that guided further purifier development. Colleagues worldwide validated purifier prototypes in simulated realistic indoor environments. Prototype products were deployed in office environments both in the United States and France. As a result of these validation studies, it was discovered that both catalyst lifetime and byproduct formation are barriers to implementing this technology. Research is ongoing at the University of Connecticut that is applicable to extending catalyst lifetime, increasing catalyst efficiency and extending activation wavelength from the ultraviolet to the visible wavelengths. It is critical that catalyst lifetime is extended to realize cost effective implementation of PCO air purification.

  2. ECONOMIC NATIONALISM’S VIABILITY UNDER GLOBALIZATION

    Directory of Open Access Journals (Sweden)

    Elena Mihaela ILIESCU

    2015-07-01

    Full Text Available The path undertaken by the world economy is irreversible - the world economic system is a system based on interdependencies, cooperation and multilateralism but economic openness is not full. Each country, in order to protect their national interest call, in different proportion, depending on the circumstances and of the economic-social and political interests, for different forms of the economic nationalism, forms that have adapted continuously to the demands required for integrating on the foreign markets. The recent global economic crisis intensified the rhetoric and the economic nationalism’s practices but it is not about rebirth, but of renewal, of remodeling the nationalist policies, globalization being a premise of the new economic nationalism. The scope of this paper is to emphasize using empirical data the fact that nationalism and globalization, from an economical point of view, are not antagonistic policies, they coexist and influence each other, both generating contradictory effects, in terms of provided opportunities and risks.

  3. The economic viability of fusion power

    International Nuclear Information System (INIS)

    Ward, D.J.; Cook, I.; Lechon, Y.; Saez, R.

    2005-01-01

    Although fusion power is being developed because of its large resource base, low environmental impact and high levels of intrinsic safety, it is important to investigate the economics of a future fusion power plant to check that the electricity produced can, in fact, have a market. The direct cost of electricity of a fusion power plant and its key dependencies on the physics and technology assumptions, are calculated, as are the materials requirements. The other important aspect of costs, the external costs which can arise from effects such as pollution, accidents and waste are also given. Fusion is found to offer the prospect of a new energy source with acceptable direct costs and very low external costs. This places fusion in a strong position in a future energy market, especially one in which environmental constraints become increasingly important

  4. Planar cell polarity gene expression correlates with tumor cell viability and prognostic outcome in neuroblastoma

    International Nuclear Information System (INIS)

    Dyberg, Cecilia; Papachristou, Panagiotis; Haug, Bjørn Helge; Lagercrantz, Hugo; Kogner, Per; Ringstedt, Thomas; Wickström, Malin; Johnsen, John Inge

    2016-01-01

    The non-canonical Wnt/Planar cell polarity (PCP) signaling pathway is a major player in cell migration during embryonal development and has recently been implicated in tumorigenesis. Transfections with cDNA plasmids or siRNA were used to increase and suppress Prickle1 and Vangl2 expression in neuroblastoma cells and in non-tumorigenic cells. Cell viability was measured by trypan blue exclusion and protein expression was determined with western blotting. Transcriptional activity was studied with luciferase reporter assay and mRNA expression with real-time RT-PCR. Immunofluorescence stainings were used to study the effects of Vangl2 overexpression in non-tumorigenic embryonic cells. Statistical significance was tested with t-test or one-way ANOVA. Here we show that high expression of the PCP core genes Prickle1 and Vangl2 is associated with low-risk neuroblastoma, suppression of neuroblastoma cell growth and decreased Wnt/β-catenin signaling. Inhibition of Rho-associated kinases (ROCKs) that are important in mediating non-canonical Wnt signaling resulted in increased expression of Prickle1 and inhibition of β-catenin activity in neuroblastoma cells. In contrast, overexpression of Vangl2 in MYC immortalized neural stem cells induced accumulation of active β-catenin and decreased the neural differentiation marker Tuj1. Similarly, genetically modified mice with forced overexpression of Vangl2 in nestin-positive cells showed decreased Tuj1 differentiation marker during embryonal development. Our experimental data demonstrate that high expression of Prickle1 and Vangl2 reduce the growth of neuroblastoma cells and indicate different roles of PCP proteins in tumorigenic cells compared to normal cells. These results suggest that the activity of the non-canonical Wnt/PCP signaling pathway is important for neuroblastoma development and that manipulation of the Wnt/PCP pathway provides a possible therapy for neuroblastoma. The online version of this article (doi:10.1186/s

  5. Present assessment of myocardial viability by nuclear imaging.

    Science.gov (United States)

    Saha, G B; MacIntyre, W J; Brunken, R C; Go, R T; Raja, S; Wong, C O; Chen, E Q

    1996-10-01

    Prospective delineation of viable from nonviable myocardium in patients with coronary artery disease in an important factor in deciding whether a patient should be revascularized or treated medically. Two common techniques--single-photon emission computed tomography (SPECT) and positron-emission computed tomography (PET)--are used in nuclear medicine using various radiopharmaceuticals for the detection of myocardial viability in patients. Thallium-201 (201Tl) and technetium-99m (99mTc)-sestamibi are the common radiopharmaceuticals used in different protocols using SPECT, whereas fluoride-18 (18F)-fluorodeoxyglucose (FDG) and rubidium-82 (82Rb) are most widely used in PET. The SPECT protocols involve stress/redistribution, stress/redistribution/reinjection, and rest/redistribution imaging techniques. Many studies have compared the results of 201Tl and (99mTc)-sestamibi SPECT with those of FDG PET; in some studies, concordant results have been found between delayed thallium and FDG results, indicating that 201Tl, although considered a perfusion agent, shows myocardial viability. Discordant results in a number of studies have been found between sestamibi and FDG, suggesting that the efficacy of sestamibi as a viability marker has yet to be established. Radiolabeled fatty acids such as iodine-123 (123I)-para-iodophenylpentadecanoic acid and carbon-11 (11C)-palmitic acid have been used for the assessment of myocardial viability with limited success. 11C-labeled acetate is a good marker of oxidative metabolism in the heart and has been used to predict the reversibility of wall motion abnormalities. (18F)-FDG is considered the marker of choice for myocardial viability, although variable results are obtained under different physiological conditions. Detection of myocardial viability can be greatly improved by developing new equipment and radiopharmaceuticals of better quality.

  6. Economic Viability of Brewery Spent Grain as a Biofuel

    Energy Technology Data Exchange (ETDEWEB)

    Morrow, Charles [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2016-01-01

    This report summarizes an investigation into the technical feasibility and economic viability of use grain wastes from the beer brewing process as fuel to generate the heat needed in subsequent brewing process. The study finds that while use of spent grain as a biofuel is technically feasible, the economics are not attractive. Economic viability is limited by the underuse of capital equipment. The investment in heating equipment requires a higher utilization that the client brewer currently anticipates. It may be possible in the future that changing factors may swing the decision to a more positive one.

  7. Radiopeptide internalisation and externalization assays: cell viability and radioligand integrity.

    Science.gov (United States)

    Naqvi, Syed Ali Raza; Sosabowski, Jane K; Nagra, Saeed Ahamad; Ishfaq, Malik M; Mather, Stephen J; Matzow, Torkjel

    2011-01-01

    Various aspects of radiopeptide receptor-mediated cell internalisation and externalization assays were assessed, including the integrity of externalized peptides and the effect of varying the pH and incubation time of the acid wash step (to remove surface receptor-bound ligand) on efficacy and cell viability. The observed intact proportion of externalized peptide was 5-10%, and acid wash buffers with pH 2.8 or below were found to be detrimental to cell viability and integrity, particularly following prolonged incubation times. Copyright © 2010 Elsevier Ltd. All rights reserved.

  8. Radiopeptide internalisation and externalisation assays: Cell viability and radioligand integrity

    International Nuclear Information System (INIS)

    Raza Naqvi, Syed Ali; Sosabowski, Jane K.; Ahamad Nagra, Saeed; Ishfaq, Malik M.; Mather, Stephen J.; Matzow, Torkjel

    2011-01-01

    Various aspects of radiopeptide receptor-mediated cell internalisation and externalisation assays were assessed, including the integrity of externalised peptides and the effect of varying the pH and incubation time of the acid wash step (to remove surface receptor-bound ligand) on efficacy and cell viability. The observed intact proportion of externalised peptide was 5-10%, and acid wash buffers with pH 2.8 or below were found to be detrimental to cell viability and integrity, particularly following prolonged incubation times.

  9. Effect of Chlorine on Giardia lamblia Cyst Viability

    OpenAIRE

    Jarroll, Edward L.; Bingham, Alan K.; Meyer, Ernest A.

    1981-01-01

    The effect of chlorine concentration on Giardia lamblia cyst viability was tested under a variety of conditions. The ability of Giardia cysts to undergo excystation was used as the criterion of viability. The experimental variables employed included temperature (25, 15, and 5°C), pH (6, 7, and 8), chlorine-cyst contact time (10, 30, and 60 min), and chlorine concentration (1 to 8 mg/liter). In the pH range studied, cyst survival generally was observed to increase as buffer pH increased. Water...

  10. Challenge testing of gametes to enhance their viability

    DEFF Research Database (Denmark)

    Callesen, Henrik

    2010-01-01

    of survival mechanism that enables them to come through the process. The details of the mechanism remain unknown but, if identified, it could have immense potential as a new way to improve the viability of embryos produced by ART. However, few publications describe systematic ways to challenge test gametes...... and then to use the results as a basis for improving gamete viability. Furthermore, new methods to monitor the reactions of gametes to such challenge tests are needed. In the present review, these two issues are discussed, as are some of the conditions necessary before a challenge test protocol can be part...

  11. Economic viability of crude glycerin in diets for lambs finished in feedlot

    Directory of Open Access Journals (Sweden)

    Mauriceia Costa Carvalho Barros

    2015-02-01

    Full Text Available The aim was to evaluate the economic viability of increasing levels of crude glycerin (CG in diets for finishing lambs. The trial was carried out at Southwest State University of Bahia, Itapetinga-BA. Twenty five crossbred, Santa Inês x Dorper, with 24 ± 2,0 kg, were housed in individual pens. The experimental design was completely randomized with five treatments and five replications. Treatments consisted of increasing levels of dietary CG (0, 2.65, 5.33, 8.06 and 10.84% and the roughage used was the Tifton 85 hay. Diets were formulated to meet the nutritional requeriments, aiming a gain of 200 g day-1. The analysis was based on use of economic indicators Net Present Value (NPV and Internal Rate of Return (IRR. The animals dry matter intake decreased linearly (P0.05 on performance and meat production results. None of the treatments showed economic viability.

  12. Effect of alcohols on filamentation, growth, viability and biofilm development in Candida albicans.

    Science.gov (United States)

    Chauhan, Nitin M; Shinde, Ravikumar B; Karuppayil, S Mohan

    2013-12-01

    In this study we report the potential of alcohols as morphogenetic regulators in Candida albicans. All the alcohols tested influenced various modes of growth like planktonic as well as biofilm forms. Viability was affected at high concentrations. Among the alcohols, the response of C. albicans to amyl alcohol (pentanol) was noteworthy. Amyl alcohol at a concentration 0.5% which was not inhibitory to growth and viability specifically inhibited morphogenetic switching from yeast to hyphal forms. It also inhibited normal biofilm development favoring yeast dominated biofilms. Based on this study we hypothesize that alcohols produced under anaerobic conditions may not favor biofilm development and support dissemination of yeast cells. Since anaerobic conditions are not found to favor production of quorum sensing molecules like farnesol, the alcohols may play a role in morphogenetic regulation.

  13. A rapid, sensitive, and cost-efficient assay to estimate viability of potato cyst nematodes.

    Science.gov (United States)

    van den Elsen, Sven; Ave, Maaike; Schoenmakers, Niels; Landeweert, Renske; Bakker, Jaap; Helder, Johannes

    2012-02-01

    Potato cyst nematodes (PCNs) are quarantine organisms, and they belong to the economically most relevant pathogens of potato worldwide. Methodologies to assess the viability of their cysts, which can contain 200 to 500 eggs protected by the hardened cuticle of a dead female, are either time and labor intensive or lack robustness. We present a robust and cost-efficient viability assay based on loss of membrane integrity upon death. This assay uses trehalose, a disaccharide present at a high concentration in the perivitelline fluid of PCN eggs, as a viability marker. Although this assay can detect a single viable egg, the limit of detection for regular field samples was higher, ≈10 viable eggs, due to background signals produced by other soil components. On the basis of 30 nonviable PCN samples from The Netherlands, a threshold level was defined (ΔA(trehalose) = 0.0094) below which the presence of >10 viable eggs is highly unlikely (true for ≈99.7% of the observations). This assay can easily be combined with a subsequent DNA-based species determination. The presence of trehalose is a general phenomenon among cyst nematodes; therefore, this method can probably be used for (for example) soybean, sugar beet, and cereal cyst nematodes as well.

  14. Novel Cell Preservation Technique to Extend Bovine In Vitro White Blood Cell Viability.

    Directory of Open Access Journals (Sweden)

    Emilie L Laurin

    Full Text Available Although cell-mediated immunity based diagnostics can be integral assays for early detection of various diseases of dairy cows, processing of blood samples for these tests is time-sensitive, often within 24 hours of collection, to maintain white blood cell viability. Therefore, to improve utility and practicality of such assays, the objective of this study was to assess the use of a novel white blood cell preservation technology in whole bovine blood. Blood samples from ten healthy cows were each divided into an unpreserved control sample and a test sample preserved with commercially-available cell transport medium. Samples were maintained at room temperature and stimulated with the mitogens pokeweed and concanavalinA, as well as with interleukin-12 p40. Stimulation was completed on days 1, 5, and 8 post-sampling. Viability of white blood cells was assessed through interferon gamma production determined with a commercial enzyme linked immunosorbent assay. In addition, mononuclear cell viability was assessed with propidium iodide flow cytometry. Greater interferon gamma production was observed on days 5 and 8 post-collection in preserved samples, with both pokeweed and concanavalinA stimulating positive interferon gamma production on day 5 post-collection. A greater proportion of the amount of interferon gamma produced on day 1 continued to be produced on days 5 and 8 post-collection with concanavalinA stimulation (with or without interleukin 12 as compared to pokeweed stimulation. Additionally, viable mononuclear cells were still present at eight days post-collection, with a higher mean proportion detected at days 5 and 8 in all stimulated preserved samples. This practical and simple method to extend in vitro white blood cell viability could benefit the efficient utilization of cell-based blood tests in ruminants.

  15. Economic viability of new launched school lunch programmes

    DEFF Research Database (Denmark)

    Jensen, Jørgen Dejgård; Smed, Sinne; Mørkbak, Morten Raun

    2013-01-01

    activities related to the schools’ support and the users’ feeling of ownership, as well as internal professionalism and leadership in the implementation of the school lunch programme are important for the viability of the programme. Strong performance on the latter factors might to some extent compensate...

  16. A comparison of assays measuring the viability of Legionella ...

    Science.gov (United States)

    Background: The relatively high prevalence of Legionella pneumophila in premise plumbing systems has been widely reported. Published reports indicate Legionella has a comparatively high resistance to chlorine and moreover has the ability to grow in phagocytic amoeba which could provide additional protection in chlorinated drinking water distribution systems. Copper-Silver (Cu-Ag) ionization treatment systems are commercially available for use in large building water systems to help control the risks from Legionella bacteria. The objectives of this study were to develop and optimize Legionella viability assays and use them to investigate the viability of Legionella bacteria after exposure to water treated with coppper and silver ions. Methods: Log phase L. pneumophila cells were used in all experiments and were generated by incubation at 35C for 48 hours in buffered yeast extract broth. Viability assays used included plating on buffered charcoal yeast extract agar to determine the number of culturable cells and treating cells with propidium monoazide (PMA) or ethidium monoazide (EMA) followed by quantitative PCR targeting mip gene of L. pneumophila. The qPCR viability assays were optimized using L. pneumophila inactivated by heat treatment at 65C for 60 min. The effectiveness of Cu-Ag ionization treatment was studied by inoculating L. pneumonia at 105 CFU/mL in water collected directly from a building water system that employed this technology and incubat

  17. Assessment of bacterial endospore viability with fluorescent dyes.

    Science.gov (United States)

    Laflamme, C; Lavigne, S; Ho, J; Duchaine, C

    2004-01-01

    To validate three fluorescence viability assays designed primarily for vegetative cells on pure Bacillus endospores. Purified fresh and gamma-irradiated Bacillus endospores (Bacillus cereus, B. coagulans and two strains of B. subtilis) were used. The viability assays were: 5-cyano-2,3-diotolyl tetrazolium chloride (CTC) to test respiratory activity and early germination, DiBAC4(3) and Live/Dead BacLight to measure membrane energization and permeabilization, respectively. Gamma irradiation treatment completely eliminated spore culturability and was used as negative control. The untreated spores showed respiratory activity after 1 h of incubation and this was characteristic of almost 100% of spores after 24 h. The membrane potential assessment gave no answer about spore viability. A lower proportion of untreated spores had permeabilized membrane compared with gamma-irradiated spores using Live/Dead BacLight (P plate count. This study shows that fluorescence tests could be applied to assess viability in potentially pathogenic Bacillus spore preparations within 1 h.

  18. The economic and social viability of Tanzanian Wildlife Management Areas

    DEFF Research Database (Denmark)

    Homewood, Katherine; Bluwstein, Jevgeniy; Lund, Jens Friis

    This policy brief contributes to assessing the economic and social viability of Tanzania’s Wildlife Management Areas (WMAs) through preliminary findings by the ‘Poverty and ecosystem Impacts of Tanzania’s Wildlife Management Areas’ (PIMA) project, focusing on benefits, costs, and their distribution...

  19. Comparison of Germination and Viability Tests for Southern Hardwood Seed

    Science.gov (United States)

    F. T. Bonner; J. L. Gammage

    1967-01-01

    This paper summarizes a 3-year evaluation of 10 methods for testing germinability and viability of the seed of six species of southern hardwood. In five of the methods, the seeds were germinated. In the others, visual, biochemical, or physical properties were the criteria. Cutting tests were best for sweetgum and Nuttall oak seed, while cutting or water germination...

  20. The effects of storage conditions on the viability of ...

    African Journals Online (AJOL)

    Long-terms recoverability of enteropathogens is necessary for future epidemiological studies to screen stool samples when conditions do not permit immediate processing. The aim of this study was to determine the viability and the recoverability of three enteropathogens bacteria (Yersinia enterocolitica, Vibrio cholerae O: 1 ...

  1. Financial viability of district mutual health insurance schemes of ...

    African Journals Online (AJOL)

    Since its implementation, unsubstantiated reports indicate increasing health care and administra-tive costs of the various DMHIS across the country without any corresponding increase in the premium level. We sought to assess the financial viability of the DMHIS in Lawra (LDMHIS) and Sissala East (SEDMHIS) districts, ...

  2. Sewage sludge pasteurization by gamma radiation: financial viability case studies

    International Nuclear Information System (INIS)

    Swinwood, J.F.; Kotler, J.

    1990-01-01

    This paper examines the financial viability of sewage sludge pasteurization by gamma radiation, by examining the following three North American scenarios: 1. Small volume sewage treatment plant experiencing high sludge disposal costs; 2. Large volume sewage treatment plant experiencing low sludge disposal costs; 3. Large volume sewage treatment plant experiencing high sludge disposal costs. (author)

  3. Sewage sludge pasteurization by gamma radiation: financial viability case studies

    Energy Technology Data Exchange (ETDEWEB)

    Swinwood, J.F.; Kotler, J. (Nordion International Inc., Kanata, Ontario (Canada))

    1990-01-01

    This paper examines the financial viability of sewage sludge pasteurization by gamma radiation, by examining the following three North American scenarios: 1. Small volume sewage treatment plant experiencing high sludge disposal costs; 2. Large volume sewage treatment plant experiencing low sludge disposal costs; 3. Large volume sewage treatment plant experiencing high sludge disposal costs. (author).

  4. Effect of Pretreatments on Seed Viability During Fruit Development ...

    African Journals Online (AJOL)

    Fiifi Baidoo

    picked from standing trees and/or forest floors, attain maximum viability and ... increase in germination potential (60%) of seeds treated with polyethylene .... Key: 60* = There was no germination 60 days after sowing; MC = Moisture content; Germ. ...... Paper presented at the pre-germplasm collection meeting on Irvingia.

  5. Effect of salt hyperosmotic stress on yeast cell viability

    Directory of Open Access Journals (Sweden)

    Logothetis Stelios

    2007-01-01

    Full Text Available During fermentation for ethanol production, yeasts are subjected to different kinds of physico-chemical stresses such as: initially high sugar concentration and low temperature; and later, increased ethanol concentrations. Such conditions trigger a series of biological responses in an effort to maintain cell cycle progress and yeast cell viability. Regarding osmostress, many studies have been focused on transcriptional activation and gene expression in laboratory strains of Saccharomyces cerevisiae. The overall aim of this present work was to further our understanding of wine yeast performance during fermentations under osmotic stress conditions. Specifically, the research work focused on the evaluation of NaCl-induced stress responses of an industrial wine yeast strain S. cerevisiae (VIN 13, particularly with regard to yeast cell growth and viability. The hypothesis was that osmostress conditions energized specific genes to enable yeast cells to survive under stressful conditions. Experiments were designed by pretreating cells with different sodium chloride concentrations (NaCl: 4%, 6% and 10% w/v growing in defined media containing D-glucose and evaluating the impact of this on yeast growth and viability. Subsequent fermentation cycles took place with increasing concentrations of D-glucose (20%, 30%, 40% w/v using salt-adapted cells as inocula. We present evidence that osmostress induced by mild salt pre-treatments resulted in beneficial influences on both cell viability and fermentation performance of an industrial wine yeast strain.

  6. The viability of business data mining in the sports environment ...

    African Journals Online (AJOL)

    Data mining can be viewed as the process of extracting previously unknown information from large databases and utilising this information to make crucial business decisions (Simoudis, 1996: 26). This paper considers the viability of using data mining tools and techniques in sports, particularly with regard to mining the ...

  7. Potential carbon credit and community expectations towards viability ...

    African Journals Online (AJOL)

    Nicholaus family

    marginal (incremental) revenues from forest carbon stock as well as the conceptual trend of forest biomass indicates that, there is ... Key words: Carbon stock payments, community preferences and REDD+ project viability. INTRODUCTION .... following criteria were used in selecting respondents especially households: 1) ...

  8. PBMC and MDAMB-231 cellular viability after telecobalt irradiation

    International Nuclear Information System (INIS)

    Andrade, Lidia M.; Campos, Tarcisio P.R.

    2002-01-01

    Radiotherapy by gamma rays are used for cancer treatment. Ionizing radiation effects on cells has been investigated. Studies in vitro were developed using tumor cell lines and ionizing radiation. The aim of this research was to test the cellular viability response of two cell types through MTT assay: human peripheral blood mononuclear cell (PBMC) and human breast carcinoma cell line MDAMB-231. These cells were irradiated with 60 Co source Theratron 80 radiotherapy machine from Atomic Energy Canada Limited with 20 x 20 cm field at 136.4 cGy/min, surface source distance 70 cm. Culture flasks contained 10 4 , 10 5 and 10 6 cells were irradiated with 100 Gy, 25 Gy, and 50 Gy using non fractionated doses. Cellular viability were evaluated 1h, 24h, 48h and 72h after irradiation and samples were measured by optical density at 595nm. Our results show that PBMC cells present lower cellular viability post irradiation. On the other hand, MDAMB-231 cells maintain viability after 50 Gy irradiation at 72h indicating cellular radioresistance. (author)

  9. Sewage sludge pasteurization by gamma radiation: Financial viability case studies

    Science.gov (United States)

    Swinwood, Jean F.; Kotler, Jiri

    This paper examines the financial viability of sewage sludge pasteurization by gamma radiation, by examining the following three North American scenarios: 1) Small volume sewage treatment plant experiencing high sludge disposal costs. 2) Large volume sewage treatment plant experiencing low sludge disposal costs. 3) Large volume sewage treatment plant experiencing high sludge disposal costs.

  10. Morphology and viability of castor bean genotypes pollen grains

    Directory of Open Access Journals (Sweden)

    Maria Selma Alves Silva Diamantino

    2016-01-01

    Full Text Available The objective of this work was to characterize the morphology and viability of the pollen of 15 genotypes of castor bean (Ricinus communis L. and to generate information that can assist in the selection of highly promising male parents for future use in genetic improvement programs aimed at producing seeds for oil extraction. Acetolysis and scanning electron microscopy was used to characterize the morphology of the pollen. The viability of the pollen grains was estimated by in vitro germination and colorimetric analysis (acetocarmine 2% and 2, 3, 5-triphenyltetrazolium chloride 1%. For the in vitro germination, pollen grains were grown in 10 types of solidified culture medium consisting of different concentrations of sucrose, boric acid, calcium nitrate, magnesium sulfate and potassium nitrate. The pollen grains had the following characteristics: medium size, isopolar and subspheroidal shape, radial symmetry, circular ambit, 3-colporate, elongated endoapertures, tectate exine and granulated sexine. The acetocarmine dye overestimated pollen viability. The media M5 and M8 were the most efficient at promoting the germination of pollen grains. The studied genotypes had high levels of viability and can therefore be used as male parents in genetic improvement programs.

  11. An evaluation of the effects of long term cryopreservation, cause of death, and time between death and donation on heart valve leaflet viability

    International Nuclear Information System (INIS)

    Strachan, K.

    1999-01-01

    The protocol for cryopreservation of allograft heart valves at the Donor Tissue Bank of Victoria was based on validation studies on the viability of the heart valve leaflets at the time of processing. The heart block is removed within 24 hours of death and the aor-tic and pulmonary valves trimmed immediately following retrieval. Following this processing, the valves are incubated in antibiotics at 30 degree C for 6 to 8 hours before being frozen in 10% DMSO at a controlled rate. A sample of tricuspid valve leaflet is placed in Krebs solution at the time of trimfning and is used for viability studies. Leaflet viability studies have been perfon-ned on all heart valves retrieved from 1993 to the present day at the Donor Tissue Bank of Victoria. Viability involves a qualitative assessment of the cellular outgrowth by leaflet fibroblasts, this assessment ranging from '-' for no outgrowth to '++++' for maximum outgrowth. Surgeons do not request valves with any particular viability and will use them whether they are viable or not. This evaluation was to determine the effects of long-term cryopreservation, cause of death, and also time lapse of heart removal following death on the viability of the retrieved leaflets. The aim of investigating the effects of long-term cryopreservation was to determine whether there was any correlation between initial viability and viability following storage for several months to several years. It was also decided to investigate whether there was any correlation between time length between death and heart retrieval and the viability. It was also thought that the cause of death may have had an effect on the viability, for example, did death by carbon monoxide poisoning have an effect on the viability of heart valve cells. Heart valves, which had been cryopreserved but could not be transplanted for various reasons were used to study the effects of cryopreservation in this study. These were thawed according to protocol and a sample of the valve

  12. Heterozygous effects of irradiated chromosomes on viability in Drosophila melanogaster

    International Nuclear Information System (INIS)

    Simmons, M.J.

    1976-01-01

    Two large experiments were conducted in order to evaluate the heterozygous effects of irradiated chromosomes on viability. Mutations were accumulated on several hundred second chromosomes by delivering doses of 2,500R over either two or four generations for total x-ray exposures of 5,000R or 10,000R. Chromosomes treated with 5,000R were screened for lethals after the first treatment, and surviving nonlethals were used to generate families of fully treated chromosomes. The members of these families shared the effects of the first irradiation, but differed with respect to those of the second. The chromosomes treated with 10,000R were not grouped into families since mutations were accumulated independently on each chromosome in that experiment. Heterozygous effects on viability of the irradiated chromosomes were tested in both isogenic (homozygous) and nonisogenic (heterozygous) genetic backgrounds. In conjunction with these tests, homozygous viabilities were determined by the marked-inversion technique. This permitted a separation of the irradiated chromosomes into those which were drastic when made homozygous and those which were not. The results indicate that drastic chromosomes have deleterious effects in heterozygous condition, since viability was reduced by 2 to 4 percent in tests performed with the 10,000R chromosomes, and by 1 percent in those involving the 5,000R material. Within a series of tests, the effects were more pronounced when the genetic background was homozygous. These results suggest that the mutants induced by high doses of x-rays are principally drastic ones which show deleterious effects on viability in heterozygous condition

  13. Non-destructive technique for determining the viability of soybean (Glycine max) seeds using FT-NIR spectroscopy.

    Science.gov (United States)

    Kusumaningrum, Dewi; Lee, Hoonsoo; Lohumi, Santosh; Mo, Changyeun; Kim, Moon S; Cho, Byoung-Kwan

    2018-03-01

    The viability of seeds is important for determining their quality. A high-quality seed is one that has a high capability of germination that is necessary to ensure high productivity. Hence, developing technology for the detection of seed viability is a high priority in agriculture. Fourier transform near-infrared (FT-NIR) spectroscopy is one of the most popular devices among other vibrational spectroscopies. This study aims to use FT-NIR spectroscopy to determine the viability of soybean seeds. Viable and artificial ageing seeds as non-viable soybeans were used in this research. The FT-NIR spectra of soybean seeds were collected and analysed using a partial least-squares discriminant analysis (PLS-DA) to classify viable and non-viable soybean seeds. Moreover, the variable importance in projection (VIP) method for variable selection combined with the PLS-DA was employed. The most effective wavelengths were selected by the VIP method, which selected 146 optimal variables from the full set of 1557 variables. The results demonstrated that the FT-NIR spectral analysis with the PLS-DA method that uses all variables or the selected variables showed good performance based on the high value of prediction accuracy for soybean viability with an accuracy close to 100%. Hence, FT-NIR techniques with a chemometric analysis have the potential for rapidly measuring soybean seed viability. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  14. Data bank for economic viability calculation of energy sources for a typical rural community at the Brazil Northern and Northeastern

    International Nuclear Information System (INIS)

    Menzel, Francine; Sabundjian, Gaiane; Vanni, Silvia Regina

    2009-01-01

    This work elaborates a data bank containing information relevant relative to energy sources in Brazil with viability and sustainability, The data bank was elaborated using the computer program Excel, where all the references are linked to the articles and to the correspondent sites. This data bank was the base for the development or the Program for the Calculation of the Economic Viability of the Alternative Energies Solar, Aeolian and Biomass (PEASEB), which results were compared to the energy generated by innovator and compact reactors (IRIS)

  15. Nucleic acid stains as indicators of Giardia muris viability following cyst inactivation.

    Science.gov (United States)

    Taghi-Kilani, R; Gyürék, L L; Millard, P J; Finch, G R; Belosevic, M

    1996-06-01

    A reliable viability assay for Giardia is required for the development of disinfection process design criteria and pathogen monitoring by water treatment utilities. Surveys of single-staining nucleic acid dyes (stain dead parasites only), and double-staining vital dye kits from Molecular Probes (stain live and dead parasites) were conducted to assess the viability of untreated, heat-killed, and chemically inactivated Giardia muris cysts. Nucleic acid staining results were compared to those of in vitro excystation and animal infectivity. Nucleic acid stain, designated as SYTO-9, was considered the best among the single-staining dyes for its ability to stain dead cysts brightly and its relatively slow decay rate of visible light emission following DNA binding. SYTO-9 staining was correlated to animal infectivity. A Live/Dead BacLight was found to be the better of 2 double-staining viability kits tested. Logarithmic survival ratios based on SYTO-9 and Live/Dead BacLight were compared to excystation and infectivity results for G. muris cysts exposed to ozone or free chlorine. The results indicate that SYTO-9 and Live/Dead BacLight staining is stable following treatment of cysts with chemical disinfectants.

  16. Optimizing conservation strategies for Mexican freetailed bats: a population viability and ecosystem services approach

    Science.gov (United States)

    Wiederholt, Ruscena; Lopez-Hoffman, Laura; Svancara, Colleen; McCracken, Gary; Thogmartin, Wayne E.; Diffendorfer, James E.; Mattson, Brady; Bagstad, Kenneth J.; Cryan, Paul; Russell, Amy; Semmens, Darius J.; Rodrigo A. Medellín,

    2015-01-01

    Conservation planning can be challenging due to the need to balance biological concerns about population viability with social concerns about the benefits biodiversity provide to society, often while operating under a limited budget. Methods and tools that help prioritize conservation actions are critical for the management of at-risk species. Here, we use a multi-attribute utility function to assess the optimal maternity roosts to conserve for maintaining the population viability and the ecosystem services of a single species, the Mexican free-tailed bat (Tadarida brasiliensis mexicana). Mexican free-tailed bats provide ecosystem services such as insect pest-suppression in agricultural areas and recreational viewing opportunities, and may be threatened by climate change and development of wind energy. We evaluated each roost based on five attributes: the maternity roost’s contribution to population viability, the pest suppression ecosystem services to the surrounding area provided by the bats residing in the roost, the ecotourism value of the roost, the risks posed to each roost structure, and the risks posed to the population of bats residing in each roost. We compared several scenarios that prioritized these attributes differently, hypothesizing that the set of roosts with the highest rankings would vary according to the conservation scenario. Our results indicate that placing higher values on different roost attributes (e.g. population importance over ecosystem service value) altered the roost rankings. We determined that the values placed on various conservation objectives are an important determinant of habitat planning.

  17. A Comparison of Three Methods for the Analysis of Skin Flap Viability: Reliability and Validity.

    Science.gov (United States)

    Tim, Carla Roberta; Martignago, Cintia Cristina Santi; da Silva, Viviane Ribeiro; Dos Santos, Estefany Camila Bonfim; Vieira, Fabiana Nascimento; Parizotto, Nivaldo Antonio; Liebano, Richard Eloin

    2018-05-01

    Objective: Technological advances have provided new alternatives to the analysis of skin flap viability in animal models; however, the interrater validity and reliability of these techniques have yet to be analyzed. The present study aimed to evaluate the interrater validity and reliability of three different methods: weight of paper template (WPT), paper template area (PTA), and photographic analysis. Approach: Sixteen male Wistar rats had their cranially based dorsal skin flap elevated. On the seventh postoperative day, the viable tissue area and the necrotic area of the skin flap were recorded using the paper template method and photo image. The evaluation of the percentage of viable tissue was performed using three methods, simultaneously and independently by two raters. The analysis of interrater reliability and viability was performed using the intraclass correlation coefficient and Bland Altman Plot Analysis was used to visualize the presence or absence of systematic bias in the evaluations of data validity. Results: The results showed that interrater reliability for WPT, measurement of PTA, and photographic analysis were 0.995, 0.990, and 0.982, respectively. For data validity, a correlation >0.90 was observed for all comparisons made between the three methods. In addition, Bland Altman Plot Analysis showed agreement between the comparisons of the methods and the presence of systematic bias was not observed. Innovation: Digital methods are an excellent choice for assessing skin flap viability; moreover, they make data use and storage easier. Conclusion: Independently from the method used, the interrater reliability and validity proved to be excellent for the analysis of skin flaps' viability.

  18. The economic and financing viability of the OPR-1000 construction at Muria Peninsula

    International Nuclear Information System (INIS)

    Moch Djoko Birmano; Mochamad Nasrullah

    2005-01-01

    The study of the economic and financing viability of the OPR-1000 construction at Muria Peninsula have been done. This study is carried out with steps as follows: updating of newest OPR-1000 technical and economic data; survey of financing sources; the calculation of electricity generation cost, electricity tariff, construction cost of OPR-1000; and calculation of OPR-1000 financing viability by using feasibility criteria such as Net Present Value (NPV), Internal Rate of Return (IRR) and capital payback period. The calculation of generation cost, electricity tariff, construction cost and financing viability criteria (IRR, NPV, Payback Period)by using KEPCO Spread Sheet. From the results of calculations how that the electricity generation cost is 4.0866 cent/kWh, while the electricity tariff is 6.6399 cent/kWh(after Value Added Tax). The total construction cost of OPR-1000 is US $4,092.09 million (included interest and financial fee, excluded initial nuclear fuel). The calculation of financing viability in base case obtained that value of IRR, NPV and Payback Period for total investment is 10.37%. US$90.52 million and 12.11 years respectively. From these results can be concluded that with the electricity tariff of 6.640 cent/kWh or 0.0664 US$/kWh, basically this project of OPR-1000 construction is very feasible and beneficial. From investment side, this project can attract investor because the rate of capital return is high enough, profit in the end of economic life is big enough and payback period is short. (author)

  19. Testing a dual-fluorescence assay to monitor the viability of filamentous cyanobacteria.

    Science.gov (United States)

    Johnson, Tylor J; Hildreth, Michael B; Gu, Liping; Zhou, Ruanbao; Gibbons, William R

    2015-06-01

    Filamentous cyanobacteria are currently being engineered to produce long-chain organic compounds, including 3rd generation biofuels. Because of their filamentous morphology, standard methods to quantify viability (e.g., plate counts) are not possible. This study investigated a dual-fluorescence assay based upon the LIVE/DEAD® BacLight™ Bacterial Viability Kit to quantify the percent viability of filamentous cyanobacteria using a microplate reader in a high throughput 96-well plate format. The manufacturer's protocol calls for an optical density normalization step to equalize the numbers of viable and non-viable cells used to generate calibration curves. Unfortunately, the isopropanol treatment used to generate non-viable cells released a blue pigment that altered absorbance readings of the non-viable cell solution, resulting in an inaccurate calibration curve. Thus we omitted this optical density normalization step, and carefully divided cell cultures into two equal fractions before the isopropanol treatment. While the resulting calibration curves had relatively high correlation coefficients, their use in various experiments resulted in viability estimates ranging from below 0% to far above 100%. We traced this to the apparent inaccuracy of the propidium iodide (PI) dye that was to stain only non-viable cells. Through further analysis via microplate reader, as well as confocal and wide-field epi-fluorescence microscopy, we observed non-specific binding of PI in viable filamentous cyanobacteria. While PI will not work for filamentous cyanobacteria, it is possible that other fluorochrome dyes could be used to selectively stain non-viable cells. This will be essential in future studies for screening mutants and optimizing photobioreactor system performance for filamentous cyanobacteria. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. An Optimized Injectable Hydrogel Scaffold Supports Human Dental Pulp Stem Cell Viability and Spreading

    Directory of Open Access Journals (Sweden)

    T. D. Jones

    2016-01-01

    Full Text Available Introduction. HyStem-C™ is a commercially available injectable hydrogel composed of polyethylene glycol diacrylate (PEGDA, hyaluronan (HA, and gelatin (Gn. These components can be mechanically tuned to enhance cell viability and spreading. Methods. The concentration of PEGDA with an added disulfide bond (PEGSSDA was varied from 0.5 to 8.0% (w/v to determine the optimal concentration for injectable clinical application. We evaluated the cell viability of human dental pulp stem cells (hDPSCs embedded in 2% (w/v PEGSSDA-HA-Gn hydrogels. Volume ratios of HA : Gn from 100 : 0 to 25 : 75 were varied to encourage hDPSC spreading. Fibronectin (Fn was added to our model to determine the effect of extracellular matrix protein concentration on hDPSC behavior. Results. Our preliminary data suggests that the hydrogel gelation time decreased as the PEGSSDA cross-linker concentration increased. The PEGSSDA-HA-Gn was biocompatible with hDPSCs, and increased ratios of HA : Gn enhanced cell viability for 14 days. Additionally, cell proliferation with added fibronectin increased significantly over time at concentrations of 1.0 and 10.0 μg/mL in PEGDA-HA-Gn hydrogels, while cell spreading significantly increased at Fn concentrations of 0.1 μg/mL. Conclusions. This study demonstrates that PEG-based injectable hydrogels maintain hDPSC viability and facilitate cell spreading, mainly in the presence of extracellular matrix (ECM proteins.

  1. VIABILITY OF SOME APPLIED COST SYSTEMS IN MANUFACTURING FIRMS: EGYPT'S CASE

    Directory of Open Access Journals (Sweden)

    Karim Mamdouh ABBAS

    2015-04-01

    Full Text Available The present study aims to analyze viability of some applied cost systems. The population of the study is some Egyptian manufacturing firms. The number of received questionnaires was 385. The results indicated that , according to the application extent indicator, the currently cost systems in Egyptian manufacturing firms may be descendingly arranged as follows: Activity Based Costing (ABC system, Target Costing (TC system , Resource Consumption Accounting (RCA system, Activity Based Management (ABM ,Other systems, Theory of Constraints (TOC and Value Chain (VC analysis.

  2. The application of flow cytometry and fluorescent probe technology for detection and assessment of viability of plant pathogenic bacteria

    NARCIS (Netherlands)

    Chitarra, L.G.; Bulk, van den R.W.

    2003-01-01

    Conventional methods to detect and assess the viability of plant pathogenic bacteria are usually based on plating assays or serological techniques. Plating assays provide information about the number of viable cells, expressed as colony-forming units, but are time-consuming and laborious.

  3. Detection of mRNA by reverse-transcription PCR as an indicator of viability in Phytophthora ramorum

    Science.gov (United States)

    A. Chimento; S.O. Cacciola; M. Garbelotto

    2011-01-01

    In the last few decades, the use of molecular tools has greatly improved the efficiency of plant disease diagnosis. However, one of the major setbacks of most molecular diagnostic approaches is their inability to differentiate between dead and viable pathogens. We propose a new strategy for the detection of plant pathogens, based on the use of mRNA as a viability...

  4. Population viability analysis of the Endangered shortnose sturgeon

    Energy Technology Data Exchange (ETDEWEB)

    Jager, Yetta [ORNL; Bevelhimer, Mark S [ORNL; Peterson, Douglas L. [University of Georgia, Athens, GA

    2011-07-01

    This study used population viability analysis (PVA) to partition the influences of potential threats to the endangered shortnose sturgeon (Acipenser brevirostrum). A workshop brought together experts to help identify potential threats including groundwater withdrawal, poor water quality, saltwater intrusion, mercury effects, harvest as by-catch, and sedimentation of spawning habitat. During the course of the project, we eliminated some threats and added new ones. Groundwater withdrawal was dismissed after a study failed to identify connection with groundwater and the majority of pumping is from a confined aquifer. We also eliminated activities on Fort Stewart as influences on spawning habitat because any successful spawning must occur upstream of Fort Stewart. We added climate change to the list of threats based on our assessment of temperature effects and expectations of sea-level rise. Our study highlighted the role of populations in nearby rivers in providing metapopulation support, raising the concern that the population in the Ogeechee River acts as a demographic sink. As part of this study, we carried out a field sampling study to analyze effects of training activities on headwater streams. We developed a new methodology for sampling design as part of this effort and used a mixed-modeling approach to identify relationships between land cover-land use, including those associated with military training activity and water quality. We found that tank training was associated with higher suspended sediment and equipment training was associated with higher organic carbon) and water quality. We detected effects of training on suspended sediment and organic carbon. We also carried out a field sampling effort in the Canoochee and Ogeechee Rivers. In the Ogeechee River, we found that dissolved oxygen in 40% of measurements during summer were below 4 mg L-1. To evaluate mercury as a potential threat, we developed a mercury uptake model and analyzed mercury levels in

  5. Viability and vigour of ageing winter wheat grains

    Directory of Open Access Journals (Sweden)

    Stanisław Grzesiuk

    2014-01-01

    Full Text Available The viability and vigour of ageing winter wheat caryopses of the cvs. Grana and Jana were tested. Viability was determined on the basis of germination capacity and rate, and vigour on the basis of the over-all activity of hydrogenases in the sprouts, exudate conductometry, analysis of sprout growth, oxygen uptake and mitochondrial protein content in the sprouts. What is called energy (or rate of germination and over-all dehydrogenase activity in embryos and sprouts and the electroconductivity of exudates were found to be very good measures of the vigour of ageing caryopses. The latter two indices of vigour should be determined at a strictly defined moment of swelling and germination. Good measures of caryopse vigour are also respiration during swelling and at the beginning of germination and mitochondrial protein content in the sprouts or seedlings. There is a high correlation between the vigour of ageing grain and its bioenergetic indices.

  6. Cosmological viability conditions for f(T) dark energy models

    Energy Technology Data Exchange (ETDEWEB)

    Setare, M.R.; Mohammadipour, N., E-mail: rezakord@ipm.ir, E-mail: N.Mohammadipour@uok.ac.ir [Department of Science, University of Kurdistan, Sanandaj (Iran, Islamic Republic of)

    2012-11-01

    Recently f(T) modified teleparallel gravity where T is the torsion scalar has been proposed as the natural gravitational alternative for dark energy. We perform a detailed dynamical analysis of these models and find conditions for the cosmological viability of f(T) dark energy models as geometrical constraints on the derivatives of these models. We show that in the phase space exists two cosmologically viable trajectory which (i) The universe would start from an unstable radiation point, then pass a saddle standard matter point which is followed by accelerated expansion de sitter point. (ii) The universe starts from a saddle radiation epoch, then falls onto the stable matter era and the system can not evolve to the dark energy dominated epoch. Finally, for a number of f(T) dark energy models were proposed in the more literature, the viability conditions are investigated.

  7. Usefulness of radionuclide scintiphotography to evaluate preserved kidney viability

    International Nuclear Information System (INIS)

    Sato, Koshi; Yokota, Kazuhiko; Uchida, Hisanori

    1987-01-01

    GAMMA imaging of the renal cortical microcirculation is a safe and non-invasive method for assessment of kidney viability before transplantation. We used trifluoperazine (TFP), urokinase and verapamil from 24 to 120 hour kidney preservation in dogs. For these preserved kidneys, we used radionuuclide scintiphotography to evaluate kidney viability. After preservation, these kidneys were perfused with technitium -99m labeled microspheres, and imaging of the renal vasculature was obtained by scintigraphy. The distribution of the microspheres was assessed visually and by computer analysis. Modified Collins' solution perfused kidneys show very poor cortical uptake with marked increase in uptake in the hilar region after preservation. In contrast, cortical flow remained relatively well preserved in kidneys perfused and preserved by use of modified Collins' solotion with TFP, urokinase and urokinase + verapamil. There was a direct correlation between these results and the capacity of kidneys treated in the same fashion to sustain life after retransplantation into the original host. (author)

  8. Nuclear Power Options Viability Study. Volume 4. Bibliography

    Energy Technology Data Exchange (ETDEWEB)

    Trauger, D B; White, J D; Sims, J W [eds.

    1986-09-01

    Documents in the Nuclear Power Options Viability Study (NPOVS) bibliography are classified under one of four headings or categories as follows: nuclear options; light water reactors; liquid metal reactors; and high temperature reactors. The collection and selection of these documents, beginning early in 1984 and continuing through March of 1986, was carried out in support of the study's objective: to explore the viabilities of several nuclear electric power generation options for commercial deployment in the United States between 2000 and 2010. There are approximately 550 articles, papers, reports, and books in the bibliography that have been selected from some 2000 surveyed. The citations have been made computer accessible to facilitate rapid on-line retrieval by keyword, author, corporate author, title, journal name, or document number.

  9. Nuclear Power Options Viability Study. Volume 4. Bibliography

    International Nuclear Information System (INIS)

    Trauger, D.B.; White, J.D.; Sims, J.W.

    1986-09-01

    Documents in the Nuclear Power Options Viability Study (NPOVS) bibliography are classified under one of four headings or categories as follows: nuclear options; light water reactors; liquid metal reactors; and high temperature reactors. The collection and selection of these documents, beginning early in 1984 and continuing through March of 1986, was carried out in support of the study's objective: to explore the viabilities of several nuclear electric power generation options for commercial deployment in the United States between 2000 and 2010. There are approximately 550 articles, papers, reports, and books in the bibliography that have been selected from some 2000 surveyed. The citations have been made computer accessible to facilitate rapid on-line retrieval by keyword, author, corporate author, title, journal name, or document number

  10. Determination of intestinal viability by Doppler ultrasonography in venous infarction.

    Science.gov (United States)

    Cooperman, M; Martin, E W; Carey, L C

    1980-01-01

    The accuracy of Doppler ultrasound in predicting the viability of ischemic intestine secondary to venous obstruction was assessed. Twenty loops of ischemic intestine were created in dogs by temporarily obstructing venous return from the bowel. Doppler arterial flow signals within the intestine quickly disappeared following venous occlusion. In ten segments, arterial signals promptly returned following release of venous occlusion. Nine of these ten segments were viable at reoperation 24 hours later. In ten segments, no arterial signals could be detected following release of venous occlusion, and only one segment proved to be viable. Doppler ultrasound findings were far more accurate in distinguishing between viable and nonviable intestine thatn were clinical guides to intestinal viability. PMID:7352777

  11. A flow cytometric method for assessing viability of intraerythrocytic hemoparasites.

    Science.gov (United States)

    Wyatt, C R; Goff, W; Davis, W C

    1991-06-24

    We have developed a rapid, reliable method of evaluating growth and viability of intraerythrocytic protozoan hemoparasites. The assay involves the selective uptake and metabolic conversion of hydroethidine to ethidium by live parasites present in intact erythrocytes. The red fluorescence imparted by ethidium intercalated into the DNA of the parasite permits the use of flow cytometry to distinguish infected erythrocytes with viable parasites from uninfected erythrocytes and erythrocytes containing dead parasites. Comparison of the fluorochromasia technique of enumerating the number and viability of hemoparasites in cultured erythrocytes with enumeration in Giemsa-stained films and uptake of [3H]hypoxanthine demonstrated the fluorochromasia technique yields comparable results. Studies with the hemoparasite, Babesia bovis, have shown the fluorochromasia technique can also be used to monitor the effect of parasiticidal drugs on parasites in vitro. The cumulative studies with the fluorochromasia assay suggest the assay will also prove useful in investigations focused on analysis of the immune response to hemoparasites and growth in vitro.

  12. Edible flowers - antioxidant activity and impact on cell viability

    OpenAIRE

    Kuceková, Zdenka; Mlček, Jiří; Humpolíček, Petr; Rop, Otakar

    2013-01-01

    The phenolic compound composition, antioxidant activity and impact on cell viability of edible flower extracts of Allium schoenoprasum; Bellis perennis; Cichorium intybus; Rumex acetosa; Salvia pratensis; Sambucus nigra; Taraxacum officinale; Tragopogon pratensis; Trifolium repens and Viola arvensis was examined for the first time. Total phenolic content of the flowers of these plants fell between 11.72 and 42.74 mg of tannin equivalents/kg of dry matter. Antioxidant activity ranged from 35.5...

  13. Effect of storage materials on viability and proximate composition of ...

    African Journals Online (AJOL)

    Viability of the G. kola seeds stored in polyethylene bag (PB) was highest (100%) followed by that of the seeds stored in fresh plantain leaf (FPL) (86.67 ± 13.33%); cement bag paper (CBP) (73.33 ± 17.64%); dry plantain leaf (DPL) (60.00%) and the control which was exposed to the ambient temperature in the laboratory ...

  14. Economic viability of biogas technology in a Bangladesh village

    International Nuclear Information System (INIS)

    Biswas, W.K.; Lucas, N.J.D.

    1997-01-01

    We estimate energy consumption for domestic cooking and biogas energy resources for 21 clusters of households in a village. Data were analyzed on a cluster basis, with investments shared. Under the present conditions, biogas technology would not be economically viable. Economic analysis involving viability tools including additional benefits of biogas technology indicate that creating a market for local biogas would make such a project feasible. (Author)

  15. Incisional abdominal hernia repair with concomitant abdominoplasty: Maintaining umbilical viability

    OpenAIRE

    Robert Phan; Elan Kaplan; Jemma K. Porrett; Yik-Hong Ho; Warren M. Rozen

    2018-01-01

    Introduction: Abdominoplasty and abdominal hernia repair are often carried out in two-stage procedures, and those describing single-stage surgery require careful dissection to preserve often only partial blood supply to the umbilicus to maintain its viability. This paper aims to describe the surgical method of laparoscopic umbilical hernia repair in association with abdominoplasty. Case presentation: A patient presents with an incisional hernia at a previous periumbilical port site of size 14...

  16. Economic viability of wind and solar energy for industrial use

    International Nuclear Information System (INIS)

    Lashkari, Z.F.

    1994-01-01

    Non conventional energy sources have begun to move from fringes of technological possibility towards commercial viability. Out of the four sources, i e. solar, wind, biogas and minimicro hydel the first two viz. wind and solar energy are of relevance for industries in western region of India. This has to be seen in the context of developments in technology and hence economics both worldwide and in India. (author)

  17. Human periodontal ligament cell viability in milk and milk substitutes.

    Science.gov (United States)

    Pearson, Robert M; Liewehr, Frederick R; West, Leslie A; Patton, William R; McPherson, James C; Runner, Royce R

    2003-03-01

    The purpose of this study was to determine the efficacy of several milk substitutes compared to whole milk in maintaining the viability of human periodontal ligament (PDL) cells on avulsed teeth. PDL cells were obtained from freshly extracted, healthy third molars and cultured in Eagle's minimal essential media (EMEM). The cells were plated onto 24-well culture plates and allowed to attach for 24 h. EMEM was replaced with refrigerated whole milk (positive control), reconstituted powdered milk, evaporated milk, or one of two baby formulas (Similac or Enfamil). Tap water served as the negative control. Tissue culture plates were incubated with the experimental media at 37 degrees C for 1, 2, 4, or 8 h. Cell viability was determined by a cell proliferation assay (CellTiter 96 AQ Assay), with absorbance read at 450 nM. A two-way ANOVA (p effect on PDL cell viability between any of the materials and whole milk. At 2 h, Enfamil and Similac performed significantly better than whole milk, whereas evaporated milk performed worse. At 4 h, Enfamil performed better than whole milk, whereas all other milk substitutes performed worse. At 8 h, all substitutes performed worse than whole milk. These results suggest that Enfamil, which is supplied in powder form that does not require special storage and has a shelf life of 18 months, is a more effective storage medium for avulsed teeth than pasteurized milk for at least 4 h.

  18. Effect of storage temperature on Streptococcus mutans viability

    Directory of Open Access Journals (Sweden)

    Ana Lídia Soares COTA

    2018-04-01

    Full Text Available Abstract Introduction Proper storage conditions and maintenance of viable biological material plays an important role in microbiological research, allowing for the opportunity to conduct future studies. Objective To evaluate the viability of Streptococcus mutans strains that were previously grown and stored under different temperatures for approximately eight years. Material and method In this study, we evaluated 393 bacterial isolates that were stored in a freezer at -80°C (G1 and 200 isolates stored in a freezer at -20°C (G2. Aliquots of each sample were plated on blood agar and mitis-salivarius bacitracin sucrose agar-solidified medium. After incubating under microaerophilic conditions in an incubator at 37°C for 72 hours, the presence, morphology and purity of bacterial growth was observed. The data were analyzed by means of descriptive statistics. Result Microbial viability was observed in almost all samples (99.7% in G1, whereas all isolates stored at -20°C were considered inviable. Conclusion The viability of S. mutans is influenced by the storage temperature of the samples, and the strains remain viable when stored under ideal temperature conditions (-80°C, even when stored for a long period of time.

  19. Ca-Lignosulphonate and sclerotial viability of Sclerotinia sclerotiorum

    Directory of Open Access Journals (Sweden)

    MATTEO MONTANARI

    2012-01-01

    Full Text Available Lignosulphonates, low cost by-products of the pulping process, have shown suppressive effects against some diseases caused by soil-borne pathogens. In this study, the effect of 1.5% v/v calcium lignosulphonate (Ca-Ls amendment to two commercial potting mixes (peat + coconut fibres; PC; and municipal compost + peat + pumice; MCPP on the viability of Sclerotinia sclerotiorum sclerotia was investigated. Sclerotia were buried in the Ca-Ls amended substrates for 30 days. Non-amended PC and MCPP, sterile sand and sterile PC with and without Ca-Ls were used as controls. The viability of sclerotia recovered from PC and MCPP amended with Ca-Ls was reduced by 50 and 42% respectively compared to control treatments. Ca-Ls amendment decreased sclerotial viability by enhancing the activity of the indigenous mycoparasitic fungi, Fusarium oxysporum, Mucor spp. and Trichoderma spp. The biocontrol ability of Ca-Ls against sclerotia was due to the stimulation of microbial activity and is, therefore, strictly dependent on the microbial composition of the substrate.

  20. Turnover of Glycerolipid Metabolite Pool and Seed Viability

    Directory of Open Access Journals (Sweden)

    Xiao-Long Hu

    2018-05-01

    Full Text Available Hydration–dehydration cycles can frequently cause stress to seeds, but can also be used to improve germination. However, the molecular basis of the stress caused is poorly understood. Herein, we examine the effects of hydration–dehydration cycles on seed viability and profile the membrane glycerolipid molecular species. We find that seed viability was not affected during the first two cycles, but significantly decreased as further cycles were applied, until all viability was lost. The abundances of seven glycerolipid classes increased and decreased through hydration and dehydration, respectively, but the phosphatidic acid and diacylglycerol abundances changed in the opposite sense, while total glycerolipid contents remained constant. This suggests that during hydration–dehydration cycles, turnover of glycerolipid metabolite pools take place, while no significant lipid synthesis or degradation is involved. As further hydration–dehydration cycles occurred, lipid unsaturation increased, plastidic lipids decreased, and phosphatidylserine acyl chains lengthened. The latter two could be lethal for seeds. Our findings reveal a novel model of membrane lipid changes, and provide new insights into the responses of seeds to hydration–dehydration cycles.

  1. Drying process strongly affects probiotics viability and functionalities.

    Science.gov (United States)

    Iaconelli, Cyril; Lemetais, Guillaume; Kechaou, Noura; Chain, Florian; Bermúdez-Humarán, Luis G; Langella, Philippe; Gervais, Patrick; Beney, Laurent

    2015-11-20

    Probiotic formulations are widely used and are proposed to have a variety of beneficial effects, depending on the probiotic strains present in the product. The impact of drying processes on the viability of probiotics is well documented. However, the impact of these processes on probiotics functionality remains unclear. In this work, we investigated variations in seven different bacterial markers after various desiccation processes. Markers were composed of four different viability evaluation (combining two growth abilities and two cytometric measurements) and in three in vitro functionalities: stimulation of IL-10 and IL-12 production by PBMCs (immunomodulation) and bacterial adhesion to hexadecane. We measured the impact of three drying processes (air-drying, freeze-drying and spray-drying), without the use of protective agents, on three types of probiotic bacteria: Bifidobacterium bifidum, Lactobacillus plantarum and Lactobacillus zeae. Our results show that the bacteria respond differently to the three different drying processes, in terms of viability and functionality. Drying methods produce important variations in bacterial immunomodulation and hydrophobicity, which are correlated. We also show that adherence can be stimulated (air-drying) or inhibited (spray-drying) by drying processes. Results of a multivariate analysis show no direct correlation between bacterial survival and functionality, but do show a correlation between probiotic responses to desiccation-rewetting and the process used to dry the bacteria. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Seca da semente de café ao sol Effect of sunlight on coffee seed viability

    Directory of Open Access Journals (Sweden)

    Oswaldo Bacchi

    1955-01-01

    Full Text Available O presente trabalho diz respeito a estudos realizados com o objetivo de observar a influência da luz solar sôbre a capacidade germinativa da semente despolpada de café e verificar, portanto, a possibilidade de se efetuar a seca desta semente exclusivamente ao sol. Além de um ensaio preliminar, foram efetuados mais outros dois, compreendendo, ao todo, duas secas ao sol, duas em um secador a baixa temperatura (±28°C, uma a sombra e uma em um secador a alta temperatura (±40°C. Pelas correlações entre as porcentagens de germinação e de umidade, que foram estabelecidas após diferentes períodos de secagem, foram tiradas as duas seguintes conclusões, que são as principais com relação ao objetivo dêstes ensaios : 1 Os raios solares são prejudiciais à capacidade germinativa da semente de café; sua nocividade acha-se, porém, intimamente ligada ao teor de umidade na semente. 2 À vista da conclusão acima, verifica-se que a semente despolpada de café e, muito provavelmente, também a não despolpada, podem ser perfeitamente sêcas a pleno sol sem que haja prejuízo para sua vitalidade, uma vez que sejam recolhidas antes que o seu teor de umidade caia abaixo do ponto crítico de 8% - 9%.In this paper the author deals with the question related to the coffee seed drying, with special attention to the effect of the sun radiations on seed viability. Based on the results obtained, the following conclusions were drawn : (a The sun rays have no specific influence on the viability of hulled coffee seed ; their effect is indirect and related to loss of moisture by the seed. (b Coffee seed can be dried in the sunlight without loss of viability if the water content does not drop under 8% to 9%. Below this critical level seed germination falls rapidly. (c The moisture content of coffee seed dried „in the sun may fall to about 7% in a short time with the consequent injury to viability. (d Under most conditions in the state of São Paulo

  3. A Luciferase Reporter Gene Assay to Measure Ebola Virus Viral Protein 35-Associated Inhibition of Double-Stranded RNA-Stimulated, Retinoic Acid-Inducible Gene 1-Mediated Induction of Interferon β.

    Science.gov (United States)

    Cannas, Valeria; Daino, Gian Luca; Corona, Angela; Esposito, Francesca; Tramontano, Enzo

    2015-10-01

    During Ebola virus (EBOV) infection, the type I interferon α/β (IFN-α/β) innate immune response is suppressed by EBOV viral protein 35 (VP35), a validated drug target. Identification of EBOV VP35 inhibitors requires a cellular system able to assess the VP35-based inhibitory functions of viral double-stranded RNA (dsRNA) IFN-β induction. We established a miniaturized luciferase gene reporter assay in A549 cells that measures IFN-β induction by viral dsRNA and is dose-dependently inhibited by VP35 expression. When compared to influenza A virus NS1 protein, EBOV VP35 showed improved inhibition of viral dsRNA-based IFN-β induction. This assay can be used to screen for EBOV VP35 inhibitors. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  4. Viability in holder of irradiated cells: distinguish between repair and cell multiplication

    International Nuclear Information System (INIS)

    Araujo, A.C. de.

    1980-01-01

    In experiments in which liquid holding recovery (LHR) was measured, the majority of cellular population is formed by non-viable cells and cell multiplication may be important for LHR expression. In order to distinguish between recuperation of viability (true LHR) and cell multiplication, it was necessary to employ improved plating techniques and a fluctuation test based on Poisson distribution. Our results are an indication that this fluctuation test, used together with the traditional method, is a good tool to distinguish repair from cell multiplication. (author)

  5. Design measures in evolutionary water cooled reactors to optimize for economic viability

    International Nuclear Information System (INIS)

    Oh, S.J.; Yu, S.K.W.; Appell, B.

    1999-01-01

    Since the mid 1980s, there have been various efforts to develop evolutionary water cooled reactors based on the current operating plant experience. To sustain and improve the economic viability, particular attention has been paid to the following aspects in developing evolutionary water cooled reactors: design simplification and increased operating margins, standardization in design as well as construction and operation, integration of operating plant insights, and consideration of safety, operability and constructability during the design stage. This paper reviews each item and discusses several examples from some of the evolutionary water cooled reactors being developed. (author)

  6. Analyze the economic and environmental viability in distributed generation of electric power from renewable sources

    International Nuclear Information System (INIS)

    Jantim Neto, Humberto

    2010-01-01

    This paper brings a brief of economical and social environmental analysis about distributed electric's energy generation, based on a comparison to centralized generation. The motivation of the proposed analysis has its origin on a reflection about politics and scheming directed to Brazilian's energy sector. This study has renewable energy resources as setting, represented for Belo Monte generation's plant and undertaking registered on the Reservation's Energy Auction 2010. The study took into account economics and technical aspects, whereas the viability analysis was formed from benefits got from different forms of electric's generation. The conclusions of this shows that distributed electric's energy generation may have economics and socio environment benefits over centralized generation. (author)

  7. Safety-related topics from the Nuclear Power Options Viability Study

    International Nuclear Information System (INIS)

    Trauger, D.B.; White, J.D.

    1986-01-01

    The Nuclear Power Options Viability Study (NPOVS) evaluated innovative reactor concepts, and this article reviews NPOVS findings, with emphasis on safety and licensing. The reactors studied were of light-water, liquid-metal, and helium-cooled concepts, and most were of modular design. Prelicensed standard plants offer an important step toward regulatory stability and early licensing approvals with public participation before major expenditures. Advanced reactors with passive safety features offer the possibility of performance-based regulation. The concepts studied appear to be potentially viable, but more complete designs will be required before economic evaluations can be definitive

  8. Comment Response on the Final Report: Peer Review of the Total System Performance Assessment-Viability Assessment (TSPA-VA)

    International Nuclear Information System (INIS)

    Pendleton, M. W.

    1999-01-01

    The Management and Operating Contractor established a Performance Assessment Peer Review Panel (hereinafter ''the Panel'') at the request of the U.S. Department of Energy Yucca Mountain Site Characterization Office. The objectives of the peer review were to provide: (1) A formal, independent evaluation and critique of Viability Assessment of a Repository at Yucca Mountain: Total System Performance Assessment, Volume 3 (DOE 1998a; hereinafter ''Total System Performance Assessment-Viability Assessment'') that was conducted in support of the Viability Assessment of a Repository at Yucca Mountain (DOE 1998b). (2) Suggestions for improvements as the U.S. Department of Energy prepares to develop the documentation for a Total System Performance Assessment to support a potential License Application. The Panel conducted a phased review over a two-year period to observe the development and, ultimately, to review the Total System Performance Assessment-Viability Assessment (DOE 1998a). During the development of the Total System Performance Assessment-Viability Assessment (DOE 1998a), the Panel submitted three Interim Reports (Whipple et al., 1997a, 1997b, and 1998) to the Management and Operating Contractor with recommendations and comments on the process models, model abstractions, and draft documentation for the Total System Performance Assessment-Viability Assessment (DOE 1998a). The Panel's Final Report Total System Performance Assessment Peer Review Panel (Whipple et al. 1999; hereinafter ''Final Report'') on the Total System Performance Assessment-Viability Assessment (DOE 1998a) is based primarily on the completed Total System Performance Assessment-Viability Assessment (DOE 1998a), the Total System Performance Assessment-Viability Assessment (TSPA-VA) Analyses Technical Basis Document (CRWMS M and O 1998), and the cited references. The Final Report (Whipple et al. 1999) includes the major points from the three Interim Reports (Whipple et al. 1997a, 1997b, and 1998

  9. Effect of surface organic coatings of cellulose nanocrystals on the viability of mammalian cell lines

    Directory of Open Access Journals (Sweden)

    Jimenez AS

    2017-09-01

    Full Text Available Ambar S Jimenez,1 Francesca Jaramillo,1 Usha D Hemraz,2 Yaman Boluk,3 Karina Ckless,1 Rajesh Sunasee1 1Department of Chemistry, State University of New York at Plattsburgh, Plattsburgh, NY, USA; 2National Research Council, Montreal, QC, Canada, 3Department of Civil & Environmental Engineering, University of Alberta and National Institute for Nanotechnology, National Research Council, Edmonton, AB, Canada Abstract: Cellulose nanocrystals (CNCs have emerged as promising candidates for a number of bio-applications. Surface modification of CNCs continues to gain significant research interest as it imparts new properties to the surface of the nanocrystals for the design of multifunctional CNCs-based materials. A small chemical surface modification can potentially lead to drastic behavioral changes of cell-material interactions thereby affecting the intended bio-application. In this work, unmodified CNCs were covalently decorated with four different organic moieties such as a diaminobutane fragment, a cyclic oligosaccharide (β-cyclodextrin, a thermoresponsive polymer (poly[N-isopropylacrylamide], and a cationic aminomethacrylamide-based polymer using different synthetic covalent methods. The effect of surface coatings of CNCs and the respective dose-response of the above organic moieties on the cell viability were evaluated on mammalian cell cultures (J774A.1 and MFC-7, using 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assays. Overall, the results indicated that cells exposed to surface-coated CNCs for 24 h did not display major changes in cell viability, membrane permeability as well as cell morphology. However, with longer exposure, all these parameters were somewhat affected, which appears not to be correlated with either anionic or cationic surface coatings of CNCs used in this study. Keywords: cellulose nanocrystals, surface coating, cell viability, MTT, LDH

  10. Determination of relative assay response factors for toxic chlorinated and brominated dioxins/furans using an enzyme immunoassay (EIA) and a chemically-activated luciferase gene expression cell bioassay (CALUX).

    Science.gov (United States)

    Samara, Fatin; Gullett, Brian K; Harrison, Robert O; Chu, Andrew; Clark, George C

    2009-04-01

    Determination of toxic activity requires knowledge of both the concentration and toxicity to evaluate the risk for adverse human health and environmental effects. A chemically-activated luciferase gene expression cell bioassay system (CALUX) and an antibody-based method enzyme immunoassay (EIA) were used to detect the dioxin-like response of several polybrominated, polychlorinated, and polybrominated/chlorinated dibenzo-p-dioxins/furans (PBDDs/Fs, PCDDs/Fs, and PBCDDs/Fs, respectively). It has been suggested that the biological activity of the brominated and mixed bromo/chloro compounds is similar to their chlorinated analogues (measured by binding to the Ah receptor). PBDD/F, PCDD/F, and PBCDD/F laboratory standards exhibited biological activity ranging over three orders of magnitude. The highest relative potency (REP) values from CALUX analysis, when compared to 2,3,7,8-TCDD, were 2,3,7,8-TBDD at 0.99 (+/-0.07), 1,2,3,7,8-PeCDD at 0.69, and 2-Br-3,7,8-TriCDD at 0.72 (+/-0.02). Cross-reactivities were calculated using EIA for several PBDDs/Fs and PBCDDs. The highest percent cross-reactivity was found for 2,3,7,8-TBDD at 138 (+/-34%), and 2,3,7-TriBDD at 84 (+/-36%).

  11. A business model design framework for viability : a business ecosystem approach

    NARCIS (Netherlands)

    D'Souza, Austin; Velthuijsen, Hugo; Wortmann, J.C.; Huitema, George

    2015-01-01

    Purpose: To facilitate the design of viable business models by proposing a novel business model design framework for viability. Design: A design science research method is adopted to develop a business model design framework for viability. The business model design framework for viability is

  12. Projecting the success of plant restoration with population viability analysis

    Science.gov (United States)

    Bell, T.J.; Bowles, M.L.; McEachern, A.K.; Brigham, C.A.; Schwartz, M.W.

    2003-01-01

    Conserving viable populations of plant species requires that they have high probabilities of long-term persistence within natural habitats, such as a chance of extinction in 100 years of less than 5% (Menges 1991, 1998; Brown 1994; Pavlik 1994; Chap. 1, this Vol.). For endangered and threatened species that have been severely reduces in range and whose habitats have been fragmented, important species conservation strategies may include augmenting existing populations or restoring new viable populations (Bowles and Whelan 1994; Chap. 2, this Vol.). Restoration objectives may include increasing population numbers to reduce extinction probability, deterministic manipulations to develop a staged cohort structure, or more complex restoration of a desired genetic structure to allow outcrossing or increase effective population size (DeMauro 1993, 1994; Bowles et al. 1993, 1998; Pavlik 1994; Knapp and Dyer 1998; Chap. 2, this Vol.). These efforts may require translocation of propagules from existing (in situ) populations, or from ex situ botanic gardens or seed storage facilities (Falk et al. 1996; Guerrant and Pavlik 1998; Chap. 2, this Vol.). Population viability analysis (PVA) can provide a critical foundation for plant restoration, as it models demographic projections used to evaluate the probability of population persistence and links plant life history with restoration strategies. It is unknown how well artificially created populations will meet demographic modeling requirements (e.g., due to artificial cohort transitions) and few, if any, PVAs have been applied to restorations. To guide application of PVA to restored populations and to illustrate potential difficulties, we examine effects of planting different life stages, model initial population sizes needed to achieve population viability, and compare demographic characteristics between natural and restored populations. We develop and compare plant population restoration viability analysis (PRVA) case studies of

  13. Viability Tests for Fresh and Stored Haemopoietic Cells

    Energy Technology Data Exchange (ETDEWEB)

    Fliedner, T. M. [Abteilung fuer klinische Physiologie, Zentrum fuer Klinische Grundlagenforschung, Universitaet Ulm, Ulm, Federal Republic of Germany (Germany)

    1969-07-15

    This paper reviews current methods of measurement of the viability of fresh and stored haemopoietic cells. The life expectancy of granulocytes and monocytes after transfusion can be studied by in-vitro labelling with {sup 3}H-DFP and subsequent autoradiography. The evaluation of data in about 30 patients with various haemopoietic conditions indicates a wide variation of the disappearance half-time of granulocytes. {sup 3}H-cytidine labels essentially all lymphocytes in vitro, predominantly in their RNA. Transfusion of {sup 3}H-cytidine-labelled lymphocytes enables one to measure the lower limit of their life-expectancy as well as their rate of RNA metabolism. If bone-marrow cells are labelled in vitro with {sup 3}H-thymidine and subsequently transfused, their capability to circulate, to reach the haemopoietic tissue of the host, to proliferate and to mature can be demonstrated. However, the repopulating capacity of frozen and thawed marrow is independent of the ability of {sup 3}H-TDR-labelled marrow cells to circulate, proliferate and mature. It is assumed that bone-marrow cells capable of repopulating depleted haemopoietic tissue are resting under steady-state conditions and can be labelled by means of {sup 3}H-TDR only using special conditions. Thus the only viability tests for fresh and stored bone-marrow cells at present appear to be bioassay methods at the animal experimental level. The results indicate the need for the development of reliable viability tests for stem cells applicable in both experimental and clinical conditions. (author)

  14. Cost Assessment Methodology and Economic Viability of Tidal Energy Projects

    Directory of Open Access Journals (Sweden)

    Eva Segura

    2017-11-01

    Full Text Available The exploitation of technologies with which to harness the energy from ocean currents will have considerable possibilities in the future thanks to their enormous potential for electricity production and their high predictability. In this respect, the development of methodologies for the economic viability of these technologies is fundamental to the attainment of a consistent quantification of their costs and the discovery of their economic viability, while simultaneously attracting investment in these technologies. This paper presents a methodology with which to determine the economic viability of tidal energy projects, which includes a technical study of the life-cycle costs into which the development of a tidal farm can be decomposed: concept and definition, design and development, manufacturing, installation, operation and maintenance and dismantling. These cost structures are additionally subdivided by considering their sub-costs and bearing in mind the main components of the tidal farm: the nacelle, the supporting tidal energy converter structure and the export power system. Furthermore, a technical study is developed in order to obtain an estimation of the annual energy produced (and, consequently, the incomes generated if the electric tariff is known by considering its principal attributes: the characteristics of the current, the ability of the device to capture energy and its ability to convert and export the energy. The methodology has been applied (together with a sensibility analysis to the particular case of a farm composed of first generation tidal energy converters in one of the Channel Island Races, the Alderney Race, in the U.K., and the results have been attained by means of the computation of engineering indexes, such as the net present value, the internal rate of return, the discounted payback period and the levelized cost of energy, which indicate that the proposed project is economically viable for all the case studies.

  15. Determination of donor corneal tissue viability by 35S incorporation

    International Nuclear Information System (INIS)

    Zlatarev, G.; Golijski, P.; Mechkarski, S.

    1978-01-01

    Effects of various storage conditions on corneal tissue viability have been studied comparatively using swine eyeballs and recording the rate of 35 S-labelled sodium sulphate binding to polysaccharides. With 6% dextran solution (Hemodex) injected into the anterior bulbus chamber of the donor, the rate of labelled compound binding remaines unaltered within the first five days, whereas with humid preservation techniques at 4 deg C binding ability is almost halved. Advantages of the proposed method of preservation are discussed with a view to clinical testing. (A.B.)

  16. Reduction of irradiated tumor cells viability under effect of hyperglycemia

    International Nuclear Information System (INIS)

    Meshcherikova, V.V.; Voloshina, E.A.

    1983-01-01

    On Ehrlich carcinoma cells adapted to growth in vivo and in vitro, cellular mechanisms of short-term hyperglycemia effect have been studied. It has been found that SH by itself leads to the loss of viability of a part of cells of ELD solid tumors manifesting during the first 24 hours upon irradiation according to the interphase death type. Tumor cell radiation injuries arising under the effect of irradiation, usually non realized up to the first division, under SH conditions potentiate its injury effect. The phenomena observed explain partially selective injury of tumoral cells in the course of irradiation under SH conditions which testifies to the prospects of its use in clinics

  17. Viability of Eggshells Ash Affecting the Setting Time of Cement

    OpenAIRE

    Fazeera Ujin; Kamran Shavarebi Ali; Zarina Yasmin Hanur Harith

    2016-01-01

    This research paper reports on the feasibility and viability of eggshells ash and its effects on the water content and setting time of cement. An experiment was carried out to determine the quantity of water required in order to follow standard cement paste of normal consistency in accordance with MS EN 196-3:2007. The eggshells ash passing the 90µm sieve was used in the investigation. Eggshells ash with percentage of 0%, 0.1%, 0.5%, 1.0%, 1.5% and 2.0% were constituted to replace the cement....

  18. Methods for isolation and viability assessment of biological organisms

    Science.gov (United States)

    Letant, Sonia Edith; Baker, Sarah Elyse; Bond, Tiziana; Chang, Allan Shih-Ping

    2015-02-03

    Isolation of biological or chemical organisms can be accomplished using a surface enhanced Raman scattering (SERS) system. The SERS system can be a single or a stacked plurality of photonic crystal membranes with noble-metal lined through pores for flowing analyte potentially containing the biological or chemical organisms. The through pores can be adapted to trap individual biological or chemical organisms and emit SERS spectra, which can then be detected by a detector and further analyzed for viability of the biological or chemical organism.

  19. Viability of human corneal keratocytes during organ culture

    DEFF Research Database (Denmark)

    Møller-Pedersen, T; Møller, H J

    1996-01-01

    The viability of human corneal keratocytes was assessed during four weeks of 'closed system' organ culture at 31 degrees C. After 28 days of culturing, the entire keratocyte population was still alive and viable because all cells incorporated uridine; a parameter for RNA-synthesis. During the first...... of keratan sulphate proteoglycan suggested that approximately 1% of the total content was lost during the period. In conclusion, our current organ culture technique can maintain a viable keratocyte population for four weeks; a viable stroma can be grafted within this period....

  20. Effect of microemulsions on cell viability of human dermal fibroblasts

    Science.gov (United States)

    Li, Juyi; Mironava, Tatsiana; Simon, Marcia; Rafailovich, Miriam; Garti, Nissim

    Microemulsions are optically clear, thermostable and isotropic mixture consisting of water, oil and surfactants. Their advantages of ease preparation, spontaneous formation, long-term stability and enhanced solubility of bioactive materials make them great potentials as vehicles in food and pharmaceutical applications. In this study, comparative in vitro cytotoxicity tests were performed to select a best formulation of microemulsion with the least toxicity for human dermal fibroblasts. Three different kinds of oils and six different kinds of surfactants were used to form microemulsions by different ratios. The effect of oil type and surfactant type as well as their proportions on cell proliferation and viability were tested.

  1. Oestrogenic activity of a textile industrial wastewater treatment plant effluent evaluated by the E-screen test and MELN gene-reporter luciferase assay

    Energy Technology Data Exchange (ETDEWEB)

    Schiliro, Tiziana, E-mail: tiziana.schiliro@unito.it [Department of Public Health and Microbiology, University of Torino, Via Santena 5bis, 10126 Torino (Italy); Porfido, Arianna [Department of Public Health and Microbiology, University of Torino, Via Santena 5bis, 10126 Torino (Italy); Spina, Federica; Varese, Giovanna Cristina [Department of Life Sciences and Systems Biology, University of Torino, Viale Mattioli 25, 10125 Torino (Italy); Gilli, Giorgio [Department of Public Health and Microbiology, University of Torino, Via Santena 5bis, 10126 Torino (Italy)

    2012-08-15

    This study quantified the biological oestrogenic activity in the effluent of a textile industrial wastewater treatment plant (IWWTP) in northwestern Italy. Samples of the IWWTP effluent were collected monthly, both before and after tertiary treatment (ozonation). After solid phase extraction, all samples were subjected to two in vitro tests of total estrogenic activity, the human breast cancer cell line (MCF-7 BUS) proliferation assay, or E-screen test, and the luciferase-transfected human breast cancer cell line (MELN) gene-reporter assay, to measure the 17{beta}-oestradiol equivalent quantity (EEQ). In the E-screen test, the mean EEQ values were 2.35 {+-} 1.68 ng/L pre-ozonation and 0.72 {+-} 0.58 ng/L post-ozonation; in the MELN gene-reporter luciferase assay, the mean EEQ values were 4.18 {+-} 3.54 ng/L pre-ozonation and 2.53 {+-} 2.48 ng/L post-ozonation. These results suggest that the post-ozonation IWWTP effluent had a lower oestrogenic activity (simple paired t-tests, p < 0.05). The average reduction of estrogenic activity of IWWTP effluent after ozonation was 67 {+-} 26% and 52 {+-} 27% as measured by E-screen test and MELN gene-reporter luciferase assay, respectively. There was a positive and significant correlation between the two tests (Rho S = 0.650, p = 0.022). This study indicates that the environmental risk is low because oestrogenic substances are deposited into the river via IWWTP at concentrations lower than those at which chronic exposure has been reported to affect the endocrine system of living organisms. -- Highlights: Black-Right-Pointing-Pointer The two in vitro tests are suited for oestrogenic activity assessment in textile WWTP. Black-Right-Pointing-Pointer There is a significant correlation between the results of the two in vitro tests. Black-Right-Pointing-Pointer The oestrogenic activity of the effluent is reduced by ozonation. Black-Right-Pointing-Pointer The input of estrogenic substances into the river via textile WWTP is low.

  2. A luciferase reporter gene assay and aryl hydrocarbon receptor 1 genotype predict the LD50 of polychlorinated biphenyls in avian species

    International Nuclear Information System (INIS)

    Manning, Gillian E.; Farmahin, Reza; Crump, Doug; Jones, Stephanie P.; Klein, Jeff; Konstantinov, Alex; Potter, Dave; Kennedy, Sean W.

    2012-01-01

    Birds differ in sensitivity to the embryotoxic effects of polychlorinated biphenyls (PCBs), which complicates environmental risk assessments for these chemicals. Recent research has shown that the identities of amino acid residues 324 and 380 in the avian aryl hydrocarbon receptor 1 (AHR1) ligand binding domain (LBD) are primarily responsible for differences in avian species sensitivity to selected dibenzo-p-dioxins and furans. A luciferase reporter gene (LRG) assay was developed in our laboratory to measure AHR1-mediated induction of a cytochrome P450 1A5 reporter gene in COS-7 cells transfected with different avian AHR1 constructs. In the present study, the LRG assay was used to measure the concentration-dependent effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and PCBs 126, 77, 105 and 118 on luciferase activity in COS-7 cells transfected with AHR1 constructs representative of 86 avian species in order to predict their sensitivity to PCB-induced embryolethality and the relative potency of PCBs in these species. The results of the LRG assay indicate that the identity of amino acid residues 324 and 380 in the AHR1 LBD are the major determinants of avian species sensitivity to PCBs. The relative potency of PCBs did not differ greatly among AHR1 constructs. Luciferase activity was significantly correlated with embryolethality data obtained from the literature (R 2 ≥ 0.87, p < 0.0001). Thus, the LRG assay in combination with the knowledge of a species' AHR1 LBD sequence can be used to predict PCB-induced embryolethality in potentially any avian species of interest without the use of lethal methods on a large number of individuals. -- Highlights: ► PCB embryolethality in birds can be predicted from a species' AHR1 genotype. ► The reporter gene assay is useful for predicting species sensitivity to PCBs. ► The relative potency of PCBs does not appear to differ between AHR1 genotypes. ► Contamination of PCB 105 and PCB 118 did not affect their relative

  3. Human CRF{sub 2} {alpha} and {beta} splice variants: pharmacological characterization using radioligand binding and a luciferase gene expression assay

    Energy Technology Data Exchange (ETDEWEB)

    Ardati, A. [Rhone-Poulenc Rorer, Cardiovascular Biology, NW4, 500 Arcola Road, Collegeville, PA (United States); Goetschy, V.; Gottowick, J.; Henriot, S.; Deuschle, U.; Kilpatrick, G.J. [Central Nervous System, Pharma Division, F. Hoffmann-La Roche AG, CH-4070 Basel (Switzerland); Valdenaire, O. [Cardiovascular Research, Pharma Division, F. Hoffmann-La Roche AG, CH-4070 Basel (Switzerland)

    1999-03-14

    Corticotropin releasing factor (CRF) receptors belong to the super-family of G protein-coupled receptors. These receptors are classified into two subtypes (CRF{sub 1} and CRF{sub 2}). Both receptors are positively coupled to adenylyl cyclase but they have a distinct pharmacology and distribution in brain. Two isoforms belonging to the CRF{sub 2} subtype receptors, CRF{sub 2{alpha}} and CRF{sub 2{beta}}, have been identified in rat and man. The neuropeptides CRF and urocortin mediate their actions through this CRF G protein-coupled receptor family. In this report, we describe the pharmacological characterization of the recently identified hCRF{sub 2{beta}} receptor. We have used radioligand binding with [{sup 125}I]-tyr{sup 0}-sauvagine and a gene expression assay in which the firefly luciferase gene expression is under the control of cAMP responsive elements. Association kinetics of [{sup 125}I]-tyr{sup 0}-sauvagine binding to the hCRF{sub 2{beta}} receptor were monophasic while dissociation kinetics were biphasic, in agreement with the kinetics results obtained with the hCRF{sub 2{alpha}} receptor. Saturation binding analysis revealed two affinity states in HEK 293 cells with binding parameters in accord with those determined kinetically and with parameters obtained with the hCRF{sub 2{alpha}} receptor. A non-hydrolysable GTP analog, Gpp(NH)p, reduced the high affinity binding of [{sup 125}I]-tyr{sup 0}-sauvagine to both hCRF{sub 2} receptor isoforms in a similar manner. The rank order of potency of CRF agonist peptides in competition experiments was identical for both hCRF{sub 2}{alpha}-helical CRF{sub (9-41)}oCRF). Similarly, agonist potency was similar for the two isoforms when studied using the luciferase gene reporter system. The peptide antagonist {alpha}-helical CRF{sub (9-41)} exhibited a non-competitive antagonism of urocortin-stimulated luciferase expression with both hCRF{sub 2} receptor isoforms. Taken together, these results indicate that the

  4. A luciferase reporter gene assay and aryl hydrocarbon receptor 1 genotype predict the LD{sub 50} of polychlorinated biphenyls in avian species

    Energy Technology Data Exchange (ETDEWEB)

    Manning, Gillian E., E-mail: gmann017@uottawa.ca [Centre for Advanced Research in Environmental Genomics, Department of Biology, University of Ottawa, Ottawa, ON, Canada K1N 6N5 (Canada); Environment Canada, National Wildlife Research Centre, Ottawa, ON, Canada K1A 0H3 (Canada); Farmahin, Reza, E-mail: mfarm070@uottawa.ca [Centre for Advanced Research in Environmental Genomics, Department of Biology, University of Ottawa, Ottawa, ON, Canada K1N 6N5 (Canada); Environment Canada, National Wildlife Research Centre, Ottawa, ON, Canada K1A 0H3 (Canada); Crump, Doug, E-mail: doug.crump@ec.gc.ca [Environment Canada, National Wildlife Research Centre, Ottawa, ON, Canada K1A 0H3 (Canada); Jones, Stephanie P., E-mail: stephanie.jones@ec.gc.ca [Environment Canada, National Wildlife Research Centre, Ottawa, ON, Canada K1A 0H3 (Canada); Klein, Jeff, E-mail: jeffery@well-labs.com [Wellington Laboratories Inc., Research Division, Guelph, ON, Canada N1G 3M5 (Canada); Konstantinov, Alex, E-mail: alex@well-labs.com [Wellington Laboratories Inc., Research Division, Guelph, ON, Canada N1G 3M5 (Canada); Potter, Dave, E-mail: dpotter@well-labs.com [Wellington Laboratories Inc., Research Division, Guelph, ON, Canada N1G 3M5 (Canada); Kennedy, Sean W., E-mail: sean.kennedy@ec.gc.ca [Centre for Advanced Research in Environmental Genomics, Department of Biology, University of Ottawa, Ottawa, ON, Canada K1N 6N5 (Canada); Environment Canada, National Wildlife Research Centre, Ottawa, ON, Canada K1A 0H3 (Canada)

    2012-09-15

    Birds differ in sensitivity to the embryotoxic effects of polychlorinated biphenyls (PCBs), which complicates environmental risk assessments for these chemicals. Recent research has shown that the identities of amino acid residues 324 and 380 in the avian aryl hydrocarbon receptor 1 (AHR1) ligand binding domain (LBD) are primarily responsible for differences in avian species sensitivity to selected dibenzo-p-dioxins and furans. A luciferase reporter gene (LRG) assay was developed in our laboratory to measure AHR1-mediated induction of a cytochrome P450 1A5 reporter gene in COS-7 cells transfected with different avian AHR1 constructs. In the present study, the LRG assay was used to measure the concentration-dependent effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and PCBs 126, 77, 105 and 118 on luciferase activity in COS-7 cells transfected with AHR1 constructs representative of 86 avian species in order to predict their sensitivity to PCB-induced embryolethality and the relative potency of PCBs in these species. The results of the LRG assay indicate that the identity of amino acid residues 324 and 380 in the AHR1 LBD are the major determinants of avian species sensitivity to PCBs. The relative potency of PCBs did not differ greatly among AHR1 constructs. Luciferase activity was significantly correlated with embryolethality data obtained from the literature (R{sup 2} ≥ 0.87, p < 0.0001). Thus, the LRG assay in combination with the knowledge of a species' AHR1 LBD sequence can be used to predict PCB-induced embryolethality in potentially any avian species of interest without the use of lethal methods on a large number of individuals. -- Highlights: ► PCB embryolethality in birds can be predicted from a species' AHR1 genotype. ► The reporter gene assay is useful for predicting species sensitivity to PCBs. ► The relative potency of PCBs does not appear to differ between AHR1 genotypes. ► Contamination of PCB 105 and PCB 118 did not affect

  5. Human islet viability and function is maintained during high density shipment in silicone rubber membrane vessels

    Science.gov (United States)

    Kitzmann, Jennifer P; Pepper, Andrew R; Lopez, Boris G; Pawlick, Rena; Kin, Tatsuya; O’Gorman, Doug; Mueller, Kathryn R; Gruessner, Angelika C; Avgoustiniatos, Efstathios S; Karatzas, Theodore; Szot, Greg L; Posselt, Andrew M; Stock, Peter G; Wilson, John R; Shapiro, AM; Papas, Klearchos K

    2014-01-01

    The shipment of human islets from processing centers to distant laboratories is beneficial for both research and clinical applications. The maintenance of islet viability and function in transit is critically important. Gas-permeable silicone rubber membrane (SRM) vessels reduce the risk of hypoxia-induced death or dysfunction during high-density islet culture or shipment. SRM vessels may offer additional advantages: they are cost-effective (fewer flasks, less labor needed), safer (lower contamination risk), and simpler (culture vessel can also be used for shipment). Human islets(IE) were isolated from two manufacturing centers and shipped in 10cm2 surface area SRM vessels in temperature and pressure controlled containers to a distant center following at least two days of culture (n = 6). Three conditions were examined: low density (LD), high density (HD), and a micro centrifuge tube negative control (NC). LD was designed to mimic the standard culture density for human islet preparations (200 IE/cm2), while HD was designed to have a 20-fold higher tissue density, which would enable the culture of an entire human isolation in 1–3 vessels. Upon receipt, islets were assessed for viability, measured by oxygen consumption rate normalized to DNA content (OCR/DNA), and quantity, measured by DNA, and, when possible, potency and function with dynamic glucose-stimulated insulin secretion (GSIS) measurements and transplants in immunodeficient B6 rag mice. Post-shipment OCR/DNA was not reduced in HD versus LD, and was substantially reduced in the NC condition. HD islets exhibited normal function post-shipment. Based on the data we conclude that entire islet isolations (up to 400,000 IE) may be shipped using a single, larger SRM vessel with no negative effect on viability and ex vivo and in vivo function. PMID:25131090

  6. Analysis of the Interactions of Botanical Extract Combinations Against the Viability of Prostate Cancer Cell Lines

    Directory of Open Access Journals (Sweden)

    Lynn S. Adams

    2006-01-01

    Full Text Available Herbal medicines are often combinations of botanical extracts that are assumed to have additive or synergistic effects. The purpose of this investigation was to compare the effect of individual botanical extracts with combinations of extracts on prostate cell viability. We then modeled the interactions between botanical extracts in combination isobolographically. Scutellaria baicalensis, Rabdosia rubescens, Panax-pseudo ginseng, Dendranthema morifolium, Glycyrrhiza uralensis and Serenoa repens were collected, taxonomically identified and extracts prepared. Effects of the extracts on cell viability were quantitated in prostate cell lines using a luminescent ATP cell viability assay. Combinations of two botanical extracts of the four most active extracts were tested in the 22Rv1 cell line and their interactions assessed using isobolographic analysis. Each extract significantly inhibited the proliferation of prostate cell lines in a time- and dose-dependent manner except repens. The most active extracts, baicalensis, D. morifolium, G. uralensis and R. rubescens were tested as two-extract combinations. baicalensis and D. morifolium when combined were additive with a trend toward synergy, whereas D. morifolium and R. rubescens together were additive. The remaining two-extract combinations showed antagonism. The four extracts together were significantly more effective than the two-by-two combinations and the individual extracts alone. Combining the four herbal extracts significantly enhanced their activity in the cell lines tested compared with extracts alone. The less predictable nature of the two-way combinations suggests a need for careful characterization of the effects of each individual herb based on their intended use.

  7. Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory.

    Science.gov (United States)

    Bowey-Dellinger, Kristen; Dixon, Luke; Ackerman, Kristin; Vigueira, Cynthia; Suh, Yewseok K; Lyda, Todd; Sapp, Kelli; Grider, Michael; Crater, Dinene; Russell, Travis; Elias, Michael; Coffield, V McNeil; Segarra, Verónica A

    2017-01-01

    Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented-one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.

  8. Economic viability of the piauçu Leporinus macrocephalus (Garavello & Britski, 1988 production

    Directory of Open Access Journals (Sweden)

    Takahashi Leonardo Susumu

    2004-01-01

    Full Text Available Brazilian fish farms presented an accelerated development during the early 90's, mainly because of the increase in fee-fishing operations. To meet the demand of this market, fish production and supply became excessive and, as a consequence, the number of fee-fishing operations, farmers and the final selling price, decreased. This study analyzes the technical aspects, production cost, profitability and economic viability of the production of piauçu (L. macrocephalus in ponds, based on information from a rural property. Feeding and fingerling costs amount to approximately 47.1% of the total production cost, representing together with the final selling price the most important factor affecting profitability. The payback period was 8.3 years, the liquid present value US$ 291.07, the internal return margin 9%, and the income-outcome ratio was 1.01, which represents an unattractive investment as a projection based on current conditions. The improvement in productive efficiency enhances the economic valuation index, and that the relative magnitude of cost and income are the most important points for the economic viability of the studied farm.

  9. Exposure to the proton scavenger glycine under alkaline conditions induces Escherichia coli viability loss.

    Directory of Open Access Journals (Sweden)

    Donna Vanhauteghem

    Full Text Available Our previous work described a clear loss of Escherichia coli (E. coli membrane integrity after incubation with glycine or its N-methylated derivatives N-methylglycine (sarcosine and N,N-dimethylglycine (DMG, but not N,N,N-trimethylglycine (betaine, under alkaline stress conditions. The current study offers a thorough viability analysis, based on a combination of real-time physiological techniques, of E. coli exposed to glycine and its N-methylated derivatives at alkaline pH. Flow cytometry was applied to assess various physiological parameters such as membrane permeability, esterase activity, respiratory activity and membrane potential. ATP and inorganic phosphate concentrations were also determined. Membrane damage was confirmed through the measurement of nucleic acid leakage. Results further showed no loss of esterase or respiratory activity, while an instant and significant decrease in the ATP concentration occurred upon exposure to either glycine, sarcosine or DMG, but not betaine. There was a clear membrane hyperpolarization as well as a significant increase in cellular inorganic phosphate concentration. Based on these results, we suggest that the inability to sustain an adequate level of ATP combined with a decrease in membrane functionality leads to the loss of bacterial viability when exposed to the proton scavengers glycine, sarcosine and DMG at alkaline pH.

  10. Exposure to the Proton Scavenger Glycine under Alkaline Conditions Induces Escherichia coli Viability Loss

    Science.gov (United States)

    Vanhauteghem, Donna; Janssens, Geert Paul Jules; Lauwaerts, Angelo; Sys, Stanislas; Boyen, Filip; Cox, Eric; Meyer, Evelyne

    2013-01-01

    Our previous work described a clear loss of Escherichia coli (E. coli) membrane integrity after incubation with glycine or its N-methylated derivatives N-methylglycine (sarcosine) and N,N-dimethylglycine (DMG), but not N,N,N-trimethylglycine (betaine), under alkaline stress conditions. The current study offers a thorough viability analysis, based on a combination of real-time physiological techniques, of E. coli exposed to glycine and its N-methylated derivatives at alkaline pH. Flow cytometry was applied to assess various physiological parameters such as membrane permeability, esterase activity, respiratory activity and membrane potential. ATP and inorganic phosphate concentrations were also determined. Membrane damage was confirmed through the measurement of nucleic acid leakage. Results further showed no loss of esterase or respiratory activity, while an instant and significant decrease in the ATP concentration occurred upon exposure to either glycine, sarcosine or DMG, but not betaine. There was a clear membrane hyperpolarization as well as a significant increase in cellular inorganic phosphate concentration. Based on these results, we suggest that the inability to sustain an adequate level of ATP combined with a decrease in membrane functionality leads to the loss of bacterial viability when exposed to the proton scavengers glycine, sarcosine and DMG at alkaline pH. PMID:23544135

  11. Improvement on The Ellis and Roberts Viability Model

    Directory of Open Access Journals (Sweden)

    Guoyan Zhou

    2016-05-01

    Full Text Available With data sets of germination percent and storage time of seed lot of wheat and sorghum stored at three different storage temperature(t, °C with three different water content (m, % of seeds, together with data set of buckwheat and lettuce reported in literatures, the possibility that seed survival curve were transformed into line by survival proportion and the relationship that logarithm of average viability period (logp50 and standard deviation of seed death distribution in time (δwith t, m and interaction between t and m were analysed. Result indicated that survival proportion transformed seed survival curve to line were much easier than the probability adopted by Ellis and Roberts, and the most important factor affecting logp50 and δ of seed lot was interaction between t and m. Thus, Ellis and Roberts viability model were suggested to be improved as Ki=Vi-p/10K-CWT (t×m to predict longevity of seed lot with initial germination percent unknown, a new model of Gi/G0=A-P/10K-CWT(t×m was constructed to predict longevity of seed lot with initial germination percent already known.

  12. A Review: The Probiotic Bacteria Viability under Different Conditions

    Directory of Open Access Journals (Sweden)

    Lavinia Florina CALINOIU

    2016-11-01

    Full Text Available This review summarized the current knowledge on probiotics and on the effects that different conditions have under this type of bacteria. The purpose of this review was to evaluate the survival rate/resistance or viability of different probiotic bacteria under several conditions, such as: processing, food composition, storage, freezing, thawing, refrigeration, temperature, oxygen, pH, gastrointestinal environment and package. Nowadays, the demand on probiotic functional foods is increasing rapidly, as the consumers became more aware about the potential health benefits, due to the fact that probiotics help in maintaining the balance and composition of intestinal flora and protect it from pathogens. A daily ingestion of 108–109 CFU ml−1 probiotic microorganisms is crucial in order to be able to demonstrate an effect in our organism, considering the dose and the effect of storage/gastrointestinal environments on the probiotic viability. Microencapsulation of probiotics in different polysaccharides was proven to be an ideal way to preserve and protect the cells from detrimental factors during processing, storage or resistance in the gastrointestinal transit, as many studies demonstrate it. There is a general interest in the improvement of the physical and mechanical stability of the polymers used in probiotics encapsulation, to ensure high population of probiotics not only in food during storage, but also after gastrointestinal digestion. Also, the carrier plays a very important role and should be carefully examined.

  13. Resuscitation at the limits of viability--an Irish perspective.

    LENUS (Irish Health Repository)

    Khan, R A

    2012-02-01

    BACKGROUND: Advances in neonatal care continue to lower the limit of viability. Decision making in this grey zone remains a challenging process. OBJECTIVE: To explore the opinions of healthcare providers on resuscitation and outcome in the less than 28-week preterm newborn. DESIGN\\/METHODS: An anonymous postal questionnaire was sent to health care providers working in maternity units in the Republic of Ireland. Questions related to neonatal management of the extreme preterm infant, and estimated survival and long-term outcome. RESULTS: The response rate was 55% (74% obstetricians and 70% neonatologists). Less than 1% would advocate resuscitation at 22 weeks, 10% of health care providers advocate resuscitation at 23 weeks gestation, 80% of all health care providers would resuscitate at 24 weeks gestation. 20% of all health care providers would advocate cessation of resuscitation efforts on 22-25 weeks gestation at 5 min of age. 65% of Neonatologists and 54% trainees in Paediatrics would cease resuscitation at 10 min of age. Obstetricians were more pessimistic about survival and long term outcome in newborns delivered between 23 and 27 weeks when compared with neonatologists. This difference was also observed in trainees in paediatrics and obstetrics. CONCLUSION: Neonatologists, trainees in paediatrics and neonatal nurses are generally more optimistic about outcome than their counterparts in obstetrical care and this is reflected in a greater willingness to provide resuscitation efforts at the limits of viability.

  14. New small molecules targeting apoptosis and cell viability in osteosarcoma.

    Directory of Open Access Journals (Sweden)

    Doris Maugg

    Full Text Available Despite the option of multimodal therapy in the treatment strategies of osteosarcoma (OS, the most common primary malignant bone tumor, the standard therapy has not changed over the last decades and still involves multidrug chemotherapy and radical surgery. Although successfully applied in many patients a large number of patients eventually develop recurrent or metastatic disease in which current therapeutic regimens often lack efficacy. Thus, new therapeutic strategies are urgently needed. In this study, we performed a phenotypic high-throughput screening campaign using a 25,000 small-molecule diversity library to identify new small molecules selectively targeting osteosarcoma cells. We could identify two new small molecules that specifically reduced cell viability in OS cell lines U2OS and HOS, but affected neither hepatocellular carcinoma cell line (HepG2 nor primary human osteoblasts (hOB. In addition, the two compounds induced caspase 3 and 7 activity in the U2OS cell line. Compared to conventional drugs generally used in OS treatment such as doxorubicin, we indeed observed a greater sensitivity of OS cell viability to the newly identified compounds compared to doxorubicin and staurosporine. The p53-negative OS cell line Saos-2 almost completely lacked sensitivity to compound treatment that could indicate a role of p53 in the drug response. Taken together, our data show potential implications for designing more efficient therapies in OS.

  15. Effects of drinking desalinated seawater on cell viability and proliferation.

    Science.gov (United States)

    Macarrão, Camila Longhi; Bachi, André Luis Lacerda; Mariano, Mario; Abel, Lucia Jamli

    2017-06-01

    Desalination of seawater is becoming an important means to address the increasing scarcity of freshwater resources in the world. Seawater has been used as drinking water in the health, food, and medical fields and various beneficial effects have been suggested, although not confirmed. Given the presence of 63 minerals and trace elements in drinking desalinated seawater (63 DSW), we evaluated their effects on the behavior of tumorigenic and nontumorigenic cells through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and annexin-V-fluorescein isothiocyanate/propidium iodide staining. Our results showed that cell viability and proliferation in the presence of 63 DSW were significantly greater than in mineral water and in the presence of fetal bovine serum in a dose-dependent manner. Furthermore, 63 DSW showed no toxic effect on murine embryonic fibroblast (NIH-3T3) and murine melanoma (B16-F10) cells. In another assay, we also showed that pre-treatment of non-adherent THP-1 cells with 63 DSW reduces apoptosis incidence, suggesting a protective effect against cell death. We conclude that cell viability and proliferation were improved by the mineral components of 63 DSW and this effect can guide further studies on health effects associated with DSW consumption.

  16. Proteus mirabilis viability after lithotripsy of struvite calculi

    Science.gov (United States)

    Prabakharan, Sabitha; Teichman, Joel M. H.; Spore, Scott S.; Sabanegh, Edmund; Glickman, Randolph D.; McLean, Robert J. C.

    2000-05-01

    Urinary calculi composed of struvite harbor urease-producing bacteria within the stone. The photothermal mechanism of holmium:YAG lithotripsy is uniquely different than other lithotripsy devices. We postulated that bacterial viability of struvite calculi would be less for calculi fragmented with holmium:YAG irradiation compared to other lithotripsy devices. Human calculi of known struvite composition (greater than 90% magnesium ammonium phosphate hexahydrate) were incubated with Proteus mirabilis. Calculi were fragmented with no lithotripsy (controls), or shock wave, intracorporeal ultrasonic, electrohydraulic, pneumatic, holmium:YAG or pulsed dye laser lithotripsy. After lithotripsy, stone fragments were sonicated and specimens were serially plated for 48 hours at 38 C. Bacterial counts and the rate of bacterial sterilization were compared. Median bacterial counts (colony forming units per ml) were 8 X 106 in controls and 3 X 106 in shock wave, 3 X 107 in ultrasonic, 4 X 105 in electrohydraulic, 8 X 106 in pneumatic, 5 X 104 in holmium:YAG and 1 X 106 in pulsed dye laser lithotripsy, p less than 0.001. The rate of bacterial sterilization was 50% for holmium:YAG lithotripsy treated stones versus 0% for each of the other cohorts, p less than 0.01. P. mirabilis viability is less after holmium:YAG irradiation compared to other lithotripsy devices.

  17. Metastasizing, Luciferase Transduced MAT-Lu Rat Prostate Cancer Models: Follow up of Bolus and Metronomic Therapy with Doxorubicin as Model Drug

    International Nuclear Information System (INIS)

    Jantscheff, Peter; Esser, Norbert; Geipel, Andreas; Woias, Peter; Ziroli, Vittorio; Goldschmidtboing, Frank; Massing, Ulrich

    2011-01-01

    The most fatal outcomes of prostate carcinoma (PCa) result from hormone-refractory variants of the tumor, especially from metastatic spread rather than from primary tumor burden. The goal of the study was to establish and apply rat MAT-Lu prostate cancer tumor models for improved non-invasive live follow up of tumor growth and metastasis by in vivo bioluminescence. We established luciferase transduced MAT-Lu rat PCa cells and studied tumor growth and metastatic processes in an ectopic as well as orthotopic setting. An intravenous bolus treatment with doxorubicin was used to demonstrate the basic applicability of in vivo imaging to follow up therapeutic intervention in these models. In vitro analysis of tissue homogenates confirmed major metastatic spread of subcutaneous tumors into the lung. Our sensitive method, however, for the first time detects metastasis also in lymph node (11/24), spleen (3/24), kidney (4/24), liver (5/24), and bone tissue (femur or spinal cord - 5/20 and 12/20, respectively). Preliminary data of orthotopic implantation (three animals) showed metastatic invasion to investigated organs in all animals but with varying preference (e.g., to lymph nodes). Intravenous bolus treatment of MAT-Lu PCa with doxorubicin reduced subcutaneous tumor growth by about 50% and the number of animals affected by metastatic lesions in lymph nodes (0/4), lung (3/6) or lumbar spine (0/2), as determined by in vivo imaging and in vitro analysis. Additionally, the possible applicability of the luciferase transduced MAT-Lu model(s) to study basic principles of metronomic therapies via jugular vein catheter, using newly established active microport pumping systems, is presented

  18. Metastasizing, Luciferase Transduced MAT-Lu Rat Prostate Cancer Models: Follow up of Bolus and Metronomic Therapy with Doxorubicin as Model Drug

    Energy Technology Data Exchange (ETDEWEB)

    Jantscheff, Peter, E-mail: jantscheff@tumorbio.uni-freiburg.de [Tumour Biology Center, Clinical Research, Department Lipids & Liposomes, Breisacher Str.117, D-79106 Freiburg (Germany); Esser, Norbert [ProQinase GmbH, Breisacher Str. 117, D-79106 Freiburg (Germany); Geipel, Andreas; Woias, Peter [Laboratory for Design of Microsystems, Department of Microsystems Engineering (IMTEK), Georges-Köhler-Allee 106, D-79110 Freiburg (Germany); Ziroli, Vittorio [Tumour Biology Center, Clinical Research, Department Lipids & Liposomes, Breisacher Str.117, D-79106 Freiburg (Germany); Goldschmidtboing, Frank [Laboratory for Design of Microsystems, Department of Microsystems Engineering (IMTEK), Georges-Köhler-Allee 106, D-79110 Freiburg (Germany); Massing, Ulrich [Tumour Biology Center, Clinical Research, Department Lipids & Liposomes, Breisacher Str.117, D-79106 Freiburg (Germany)

    2011-06-17

    The most fatal outcomes of prostate carcinoma (PCa) result from hormone-refractory variants of the tumor, especially from metastatic spread rather than from primary tumor burden. The goal of the study was to establish and apply rat MAT-Lu prostate cancer tumor models for improved non-invasive live follow up of tumor growth and metastasis by in vivo bioluminescence. We established luciferase transduced MAT-Lu rat PCa cells and studied tumor growth and metastatic processes in an ectopic as well as orthotopic setting. An intravenous bolus treatment with doxorubicin was used to demonstrate the basic applicability of in vivo imaging to follow up therapeutic intervention in these models. In vitro analysis of tissue homogenates confirmed major metastatic spread of subcutaneous tumors into the lung. Our sensitive method, however, for the first time detects metastasis also in lymph node (11/24), spleen (3/24), kidney (4/24), liver (5/24), and bone tissue (femur or spinal cord - 5/20 and 12/20, respectively). Preliminary data of orthotopic implantation (three animals) showed metastatic invasion to investigated organs in all animals but with varying preference (e.g., to lymph nodes). Intravenous bolus treatment of MAT-Lu PCa with doxorubicin reduced subcutaneous tumor growth by about 50% and the number of animals affected by metastatic lesions in lymph nodes (0/4), lung (3/6) or lumbar spine (0/2), as determined by in vivo imaging and in vitro analysis. Additionally, the possible applicability of the luciferase transduced MAT-Lu model(s) to study basic principles of metronomic therapies via jugular vein catheter, using newly established active microport pumping systems, is presented.

  19. Metastasizing, Luciferase Transduced MAT‑Lu Rat Prostate Cancer Models: Follow up of Bolus and Metronomic Therapy with Doxorubicin as Model Drug

    Directory of Open Access Journals (Sweden)

    Peter Woias

    2011-06-01

    Full Text Available The most fatal outcomes of prostate carcinoma (PCa result from hormone-refractory variants of the tumor, especially from metastatic spread rather than from primary tumor burden. The goal of the study was to establish and apply rat MAT-Lu prostate cancer tumor models for improved non-invasive live follow up of tumor growth and metastasis by in vivo bioluminescence. We established luciferase transduced MAT-Lu rat PCa cells and studied tumor growth and metastatic processes in an ectopic as well as orthotopic setting. An intravenous bolus treatment with doxorubicin was used to demonstrate the basic applicability of in vivo imaging to follow up therapeutic intervention in these models. In vitro analysis of tissue homogenates confirmed major metastatic spread of subcutaneous tumors into the lung. Our sensitive method, however, for the first time detects metastasis also in lymph node (11/24, spleen (3/24, kidney (4/24, liver (5/24, and bone tissue (femur or spinal cord - 5/20 and 12/20, respectively. Preliminary data of orthotopic implantation (three animals showed metastatic invasion to investigated organs in all animals but with varying preference (e.g., to lymph nodes. Intravenous bolus treatment of MAT-Lu PCa with doxorubicin reduced subcutaneous tumor growth by about 50% and the number of animals affected by metastatic lesions in lymph nodes (0/4, lung (3/6 or lumbar spine (0/2, as determined by in vivo imaging and in vitro analysis. Additionally, the possible applicability of the luciferase transduced MAT-Lu model(s to study basic principles of metronomic therapies via jugular vein catheter, using newly established active microport pumping systems, is presented.

  20. Disruption of bbe02 by Insertion of a Luciferase Gene Increases Transformation Efficiency of Borrelia burgdorferi and Allows Live Imaging in Lyme Disease Susceptible C3H Mice.

    Directory of Open Access Journals (Sweden)

    Kamfai Chan

    Full Text Available Lyme disease is the most prevalent tick-borne disease in North America and Europe. The causative agent, Borrelia burgdorferi persists in the white-footed mouse. Infection with B. burgdorferi can cause acute to persistent multisystemic Lyme disease in humans. Some disease manifestations are also exhibited in the mouse model of Lyme disease. Genetic manipulation of B. burgdorferi remains difficult. First, B. burgdorferi contains a large number of endogenous plasmids with unique sequences encoding unknown functions. The presence of these plasmids needs to be confirmed after each genetic manipulation. Second, the restriction modification defense systems, including that encoded by bbe02 gene lead to low transformation efficiency in B. burgdorferi. Therefore, studying the molecular basis of Lyme pathogenesis is a challenge. Furthermore, investigation of the role of a specific B. burgdorferi protein throughout infection requires a large number of mice, making it labor intensive and expensive. To overcome the problems associated with low transformation efficiency and to reduce the number of mice needed for experiments, we disrupted the bbe02 gene of a highly infectious and pathogenic B. burgdorferi strain, N40 D10/E9 through insertion of a firefly luciferase gene. The bbe02 mutant shows higher transformation efficiency and maintains luciferase activity throughout infection as detected by live imaging of mice. Infectivity and pathogenesis of this mutant were comparable to the wild-type N40 strain. This mutant will serve as an ideal parental strain to examine the roles of various B. burgdorferi proteins in Lyme pathogenesis in the mouse model in the future.

  1. The location and nature of general anesthetic binding sites on the active conformation of firefly luciferase; a time resolved photolabeling study.

    Directory of Open Access Journals (Sweden)

    Sivananthaperumal Shanmugasundararaj

    Full Text Available Firefly luciferase is one of the few soluble proteins that is acted upon by a wide variety of general anesthetics and alcohols; they inhibit the ATP-driven production of light. We have used time-resolved photolabeling to locate the binding sites of alcohols during the initial light output, some 200 ms after adding ATP. The photolabel 3-azioctanol inhibited the initial light output with an IC50 of 200 µM, close to its general anesthetic potency. Photoincorporation of [(3H]3-azioctanol into luciferase was saturable but weak. It was enhanced 200 ms after adding ATP but was negligible minutes later. Sequencing of tryptic digests by HPLC-MSMS revealed a similar conformation-dependence for photoincorporation of 3-azioctanol into Glu-313, a residue that lines the bottom of a deep cleft (vestibule whose outer end binds luciferin. An aromatic diazirine analog of benzyl alcohol with broader side chain reactivity reported two sites. First, it photolabeled two residues in the vestibule, Ser-286 and Ile-288, both of which are implicated with Glu-313 in the conformation change accompanying activation. Second, it photolabeled two residues that contact luciferin, Ser-316 and Ser-349. Thus, time resolved photolabeling supports two mechanisms of action. First, an allosteric one, in which anesthetics bind in the vestibule displacing water molecules that are thought to be involved in light output. Second, a competitive one, in which anesthetics bind isosterically with luciferin. This work provides structural evidence that supports the competitive and allosteric actions previously characterized by kinetic studies.

  2. Using a split luciferase assay (SLA) to measure the kinetics of cell-cell fusion mediated by herpes simplex virus glycoproteins.

    Science.gov (United States)

    Saw, Wan Ting; Matsuda, Zene; Eisenberg, Roselyn J; Cohen, Gary H; Atanasiu, Doina

    2015-11-15

    Herpes simplex virus (HSV) entry and cell-cell fusion require the envelope proteins gD, gH/gL and gB. We propose that receptor-activated conformational changes to gD activate gH/gL, which then triggers gB (the fusogen) into an active form. To study this dynamic process, we have adapted a dual split protein assay originally developed to study the kinetics of human immunodeficiency virus (HIV) mediated fusion. This assay uses a chimera of split forms of renilla luciferase (RL) and green fluorescent protein (GFP). Effector cells are co-transfected with the glycoproteins and one of the split reporters. Receptor-bearing target cells are transfected with the second reporter. Co-culture results in fusion and restoration of RL, which can convert a membrane permeable substrate into a luminescent product, thereby enabling one to monitor initiation and extent of fusion in live cells in real time. Restoration of GFP can also be studied by fluorescence microscopy. Two sets of split reporters have been developed: the original one allows one to measure fusion kinetics over hours whereas the more recent version was designed to enhance the sensitivity of RL activity allowing one to monitor both initiation and rates of fusion in minutes. Here, we provide a detailed, step-by-step protocol for the optimization of the assay (which we call the SLA for split luciferase assay) using the HSV system. We also show several examples of the power of this assay to examine both the initiation and kinetics of cell-cell fusion by wild type forms of gD, gB, gH/gL of both serotypes of HSV as well as the effect of mutations and antibodies that alter the kinetics of fusion. The SLA can be applied to other viral systems that carry out membrane fusion. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Evaluation of goat milk as storage media to preserve viability of human periodontal ligament cells in vitro.

    Science.gov (United States)

    Ulusoy, Ayça Tuba; Kalyoncuoglu, Elif; Kaya, Senay; Cehreli, Zafer Cavit

    2016-08-01

    The purpose of this study was to evaluate the effectiveness of goat milk as a storage media for maintenance of periodontal ligament (PDL) cell viability of avulsed teeth and compare it with commonly used and/or investigated storage media. PDL cells were obtained from the root surface of healthy premolars and were cultured in Eagle's maintenance medium (EMM). Cell cultures were treated with the following storage media: tap water (negative control); EMM (positive control); Hank's balanced salt solution; ultra high temperature (UHT) long-shelf-life lactose-free cow milk; UHT long-shelf-life whole cow milk; UHT long-shelf-life skimmed cow milk; UHT long-shelf-life soy milk; UHT long-shelf-life goat milk, UHT long-shelf-life follow on milk with probiotic, 20% propolis, and egg white. Culture plates were incubated with experimental media at 20°C for 1, 3, 6, 12, and 24 h. PDL cell viability was assessed by tetrazolium salt-based colorimetric (MTT) assay at each test period. One-way anova was used to evaluate the effects of storage solutions at each time point, followed by post hoc Duncan's multiple comparison test (P = 0.05). A dendrogram was constructed to show the arrangement of hierarchical clustering. Goat milk displayed the highest capacity to maintain cell viability at all test intervals (P milk with the probiotic showed the lowest time-dependent PDL cell viability among all test media (P milks, HBSS performed significantly less effectively in maintaining PDL cell viability during the entire test period (P milk can be recommended as a suitable storage medium for avulsed teeth. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  4. Ovulation order mediates a trade-off between pre-hatching and post-hatching viability in an altricial bird.

    Directory of Open Access Journals (Sweden)

    Keith W Sockman

    2008-03-01

    Full Text Available Simultaneously dependent siblings often compete for parentally provided resources. This competition may lead to mortality, the probability of which may be a function, in part, of the individual offspring's production order. In birds, serial ovulation followed by hatching asynchrony of simultaneous dependents leads to differences in post-hatching survival that largely depend on ovulation (laying order. This has led to the widespread assumption that early-laid eggs are of greater value and therefore should possess different maternally manipulated characteristics than later-laid eggs. However, this perspective ignores the potential effect of laying order on pre-hatching viability, an effect which some studies suggest should offset the effect of laying order on post-hatching viability. I examined the relationship between laying order and hatching and fledging probability in wild, free-living Lincoln's sparrows (Melospiza lincolnii. In broods with complete hatching success, first-laid and therefore first-hatched offspring had the highest probability of fledging, and fledging probability declined with increasing laying order. However, first-laid eggs were less likely than later-laid eggs to hatch. This effect of laying order on pre-hatching viability seemed to offset that on post-hatching viability, and, consistently, maternal investment in egg size varied little if at all with respect to laying order. These results suggest that ovulation order mediates a trade-off between pre-hatching and post-hatching viability and should encourage a re-evaluation of the solitary role post-embryonic survival often plays when researchers make assumptions about the value of propagules based on the order in which they are produced.

  5. The relationship between myocardial blood flow and myocardial viability after reperfusion. Myocardial viability assessed by 15O-water-PET

    International Nuclear Information System (INIS)

    Tsukagoshi, Joichi

    1994-01-01

    The purpose of this study was to examine the relationship between myocardial blood flow and myocardial viability in the ischemic canine myocardium after reperfusion. Transient ischemia was induced by 60-, 90-, and 180-minute occlusion of the left anterior descending coronary artery. Myocardial blood flow (MBF) was measured in the areas in which regional contractility was severely impaired (ehocardiographically akinetic or dyskinetic) in the early reperfusion period by 15 O-water positron emission tomography (PET) 12 hours and 4 weeks after reperfusion. An MBF ratio of ischemic to nonischemic regions 12 hours after reperfusion was inversely correlated with the amount of histologically determined tissue necrosis (r=-0.74). The regional contractility recovered 4 weeks later in the areas where an MBF ratio was 0.48 or greater, but did not recover in the areas with a lower MBF ratio. Thus, myocardial viability can be appropriately predicted in the early phase of myocardial perfusion by PET with 15 O-water even in the absence of metabolic imaging. (author)

  6. Rapid assessment of repair of ultraviolet DNA damage with a modified host-cell reactivation assay using a luciferase reporter gene and correlation with polymorphisms of DNA repair genes in normal human lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Qiao Yawei; Spitz, Margaret R.; Guo Zhaozheng; Hadeyati, Mohammad; Grossman, Lawrence; Kraemer, Kenneth H.; Wei Qingyi

    2002-11-30

    As DNA repair plays an important role in genetic susceptibility to cancer, assessment of the DNA repair phenotype is critical for molecular epidemiological studies of cancer. In this report, we compared use of the luciferase (luc) reporter gene in a host-cell reactivation (HCR) (LUC) assay of repair of ultraviolet (UV) damage to DNA to use of the chloramphenicol (cat) gene-based HCR (CAT) assay we used previously for case-control studies. We performed both the assays on cryopreserved lymphocytes from 102 healthy non-Hispanic white subjects. There was a close correlation between DNA repair capacity (DRC) as measured by the LUC and CAT assays. Although these two assays had similar variation, the LUC assay was faster and more sensitive. We also analyzed the relationship between DRC and the subjects' previously determined genotypes for four polymorphisms of two nucleotide-excision repair (NER) genes (in intron 9 of xeroderma pigmentosum (XP) C and exons 6, 10 and 23 of XPD) and one polymorphism of a base-excision repair gene in exon 10 of X-ray complementing group 1 (XRCC1). The DRC was significantly lower in subjects homozygous for one or more polymorphisms of the two NER genes than in subjects with other genotypes (P=0.010). In contrast, the polymorphic XRCC1 allele had no significant effect on DRC. These results suggest that the post-UV LUC assay measures NER phenotype and that polymorphisms of XPC and XPD genes modulate DRC. For population studies of the DNA repair phenotype, many samples need to be evaluated, and so the LUC assay has several advantages over the CAT assay: the LUC assay was more sensitive, had less variation, was not radioactive, was easier to perform, and required fewer cryopreserved cells. These features make the LUC-based HCR assay suitable for molecular epidemiological studies.

  7. Cytotoxicity and Effects on Cell Viability of Nickel Nanowires

    KAUST Repository

    Rodriguez, Jose E.

    2013-05-01

    Recently, magnetic nanoparticles are finding an increased use in biomedical applications and research. Nanobeads are widely used for cell separation, biosensing and cancer therapy, among others. Due to their properties, nanowires (NWs) are gaining ground for similar applications and, as with all biomaterials, their cytotoxicity is an important factor to be considered before conducting biological studies with them. In this work, the cytotoxic effects of nickel NWs (Ni NWs) were investigated in terms of cell viability and damage to the cellular membrane. Ni NWs with an average diameter of 30-34 nm were prepared by electrodeposition in nanoporous alumina templates. The templates were obtained by a two-step anodization process with oxalic acid on an aluminum substrate. Characterization of NWs was done using X-Ray diffraction (XRD) and energy dispersive X-Ray analysis (EDAX), whereas their morphology was observed with scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Cell viability studies were carried out on human colorectal carcinoma cells HCT 116 by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) cell proliferation colorimetric assay, whereas the lactate dehydrogenase (LDH) homogenous membrane fluorimetric assay was used to measure the degree of cell membrane rupture. The density of cell seeding was calculated to obtain a specific cell number and confluency before treatment with NWs. Optical readings of the cell-reduced MTT products were measured at 570 nm, whereas fluorescent LDH membrane leakage was recorded with an excitation wavelength of 525 nm and an emission wavelength of 580 - 640 nm. The effects of NW length, cell exposure time, as well as NW:cell ratio, were evaluated through both cytotoxic assays. The results show that cell viability due to Ni NWs is affected depending on both exposure time and NW number. On the other hand, membrane rupture and leakage was only significant at later exposure times. Both

  8. Evaluation of implementation viability gap funding (VGF) policy on toll road investment in Indonesia

    Science.gov (United States)

    Mahani, Iris; Tamin, Rizal Z.; Pribadi, Krishna S.; Wibowo, Andreas

    2017-11-01

    VGF policy for toll road investment in Indonesia must be reviewed. Since 2012 the Government of Indonesia (GOI) has issued viability gap funding (VGF) policy for PPP infrastructure project through ministry of finance decision (PMK) No.223/2012. One of VGF purpose is to improve the financial feasibility. In the toll road investment in Indonesia, the implementation of this policy has some problems. This study aimed to evaluate the policy by seeking implementation constraints so can be given an alternative. This research was conducted qualitatively, included aspects of implementation process VGF policy. The analysis process is based on literature study and in-depth interviews to related parties include business entity, ministry of finance, and the ministry of public works, Indonesia Toll Road Authority (BPJT) and professional societies. The literature review conducted by reviewing existing policies and best practices in countries that already practice VGF. The conclusion of this study are 1) There is a conflict of regulation in viability gap funding (VGF) for toll road investment in Indonesia; 2) If Government of Indonesia (GOI) want implement construction grant as VGF, so the regulation must improve in time limited for submission and clearly define limited given in regulation; 3) If GOI want implement partial construction as VGF, so the regulation must be improve in guideline for submission and given.

  9. Succinate modulates Ca(2+) transient and cardiomyocyte viability through PKA-dependent pathway.

    Science.gov (United States)

    Aguiar, Carla J; Andrade, Vanessa L; Gomes, Enéas R M; Alves, Márcia N M; Ladeira, Marina S; Pinheiro, Ana Cristina N; Gomes, Dawidson A; Almeida, Alvair P; Goes, Alfredo M; Resende, Rodrigo R; Guatimosim, Silvia; Leite, M Fatima

    2010-01-01

    GPR91 is an orphan G-protein-coupled receptor (GPCR) that has been characterized as a receptor for succinate, a citric acid cycle intermediate, in several tissues. In the heart, the role of succinate is unknown. We now report that rat ventricular cardiomyocytes express GPR91. We found that succinate, through GPR91, increases the amplitude and the rate of decline of global Ca(2+) transient, by increasing the phosphorylation levels of ryanodine receptor and phospholamban, two well known Ca(2+) handling proteins. The effects of succinate on Ca(2+) transient were abolished by pre-treatment with adenylyl cyclase and cAMP-dependent protein kinase (PKA) inhibitors. Direct PKA activation by succinate was further confirmed using a FRET-based A-kinase activity reporter. Additionally, succinate decreases cardiomyocyte viability through a caspase-3 activation pathway, effect also prevented by PKA inhibition. Taken together, these observations show that succinate acts as a signaling molecule in cardiomyocytes, modulating global Ca(2+) transient and cell viability through a PKA-dependent pathway. 2009 Elsevier Ltd. All rights reserved.

  10. Evaluation of Periodontal Ligament Cell Viability in Three Different Storage Media: An in Vitro Study

    Directory of Open Access Journals (Sweden)

    Meenakshi Sharma

    2016-01-01

    Full Text Available Objectives: This study was undertaken to evaluate the viability of periodontal ligament (PDL cells of avulsed teeth in three different storage media.Materials and Methods: Forty-five premolars extracted for orthodontic therapeutic purposes were randomly and equally divided into three groups based on storage media used [Group I: milk (control; Group II: aloe vera (experimental; Group III: egg white (experimental]. Following extractions, the teeth were placed in one of the three different storage media for 30 minutes, following which the scrapings of the PDL from these teeth were collected in Falcon tubes containing collagenase enzyme in 2.5 mL of phosphate buffered saline. The tubes were subsequently incubated for 30 minutes and centrifuged for five minutes at 800 rpm. The obtained PDL cells were stained with Trypan Blue and were observed under optical microscope. The percentage of viable cells was calculated.Results: Aloe vera showed the highest percentage of viable cells (114.3±8.0, followed by egg white (100.9±6.3 and milk (101.1±7.3.Conclusion: Within the limitations of this study, it appears that aloe vera maintains PDL cell viability better than egg white or milk.

  11. Repeatability and reproducibility of Population Viability Analysis (PVA and the implications for threatened species management

    Directory of Open Access Journals (Sweden)

    Clare Morrison

    2016-08-01

    Full Text Available Conservation triage focuses on prioritizing species, populations or habitats based on urgency, biodiversity benefits, recovery potential as well as cost. Population Viability Analysis (PVA is frequently used in population focused conservation prioritizations. The critical nature of many of these management decisions requires that PVA models are repeatable and reproducible to reliably rank species and/or populations quantitatively. This paper assessed the repeatability and reproducibility of a subset of previously published PVA models. We attempted to rerun baseline models from 90 publicly available PVA studies published between 2000-2012 using the two most common PVA modelling software programs, VORTEX and RAMAS-GIS. Forty percent (n = 36 failed, 50% (45 were both repeatable and reproducible, and 10% (9 had missing baseline models. Repeatability was not linked to taxa, IUCN category, PVA program version used, year published or the quality of publication outlet, suggesting that the problem is systemic within the discipline. Complete and systematic presentation of PVA parameters and results are needed to ensure that the scientific input into conservation planning is both robust and reliable, thereby increasing the chances of making decisions that are both beneficial and defensible. The implications for conservation triage may be far reaching if population viability models cannot be reproduced with confidence, thus undermining their intended value.

  12. Gamma Irradiation Effects on Oil Palm (Elaies Guineensis) Pollen Viability, Fruits and Bunch Formations

    International Nuclear Information System (INIS)

    Aida Nazlyn Nazari; Azhar Mohamad; Shuhaimi Shamsuddin

    2014-01-01

    Assessing performance and genetic diversity of the wild material of oil palm is important for understanding genetic structure of natural oil palm populations towards improvement of the crops. This information is important for oil palm breeding programs, and also for continued ex-situ conservation of the germplasm and breeding program in Malaysia. Mutation induction is one of the alternative ways in creating variants for selection in the breeding program. In this study, evaluation on the effect of irradiated pollen towards pollen viability, bunches formation and no. of parthenocarpic fruits were conducted. Series of acute gamma radiation at dose 0, 10, 20, 40, 50, 100, 200, 300, 500, 100 and 2000 Gy were exposed to Elaies guineensis pollen. Increasing level of irradiation higher than 200 Grays (Gy) affects pollen viability based on the 12 consecutive days observation evaluated in situ conditions. Besides, at this level of irradiation (> 200 Gy), the pollen tube formation were disrupted and subsequently unable the pollen to reach the ovule which cause the embryo to aborted and contributed to the formation of parthenocarpic fruits and rotten bunches. These observations suggested that at low levels of irradiation (< 200 Gy) may damage only part of the generative nucleus while maintaining its capacity to fertilise the egg cells and lead to hybridization. (author)

  13. Toward an integrated approach to nutritional quality, environmental sustainability, and economic viability: research and measurement gaps.

    Science.gov (United States)

    Herforth, Anna; Frongillo, Edward A; Sassi, Franco; Mclean, Mireille Seneclauze; Arabi, Mandana; Tirado, Cristina; Remans, Roseline; Mantilla, Gilma; Thomson, Madeleine; Pingali, Prabhu

    2014-12-01

    Nutrition is affected by numerous environmental and societal causes. This paper starts with a simple framework based on three domains: nutritional quality, economic viability, and environmental sustainability, and calls for an integrated approach in research to simultaneously account for all three. It highlights limitations in the current understanding of each domain, and how they influence one another. Five research topics are identified: measuring the three domains (nutritional quality, economic viability, environmental sustainability); modeling across disciplines; furthering the analysis of food systems in relation to the three domains; connecting climate change and variability to nutritional quality; and increasing attention to inequities among population groups in relation to the three domains. For an integrated approach to be developed, there is a need to identify and disseminate available metrics, modeling techniques, and tools to researchers, practitioners, and policy makers. This is a first step so that a systems approach that takes into account potential environmental and economic trade-offs becomes the norm in analyzing nutrition and food-security patterns. Such an approach will help fill critical knowledge gaps and will guide researchers seeking to define and address specific research questions in nutrition in their wider socioeconomic and environmental contexts. © 2014 New York Academy of Sciences.

  14. Different methods to quantify Listeria monocytogenesbiofilms cells showed different profile in their viability

    Directory of Open Access Journals (Sweden)

    Lizziane Kretli Winkelströter

    2015-03-01

    Full Text Available Listeria monocytogenes is a foodborne pathogen able to adhere and to form biofilms in several materials commonly present in food processing plants. The aim of this study was to evaluate the resistance of Listeria monocytogenes attached to abiotic surface, after treatment with sanitizers, by culture method, microscopy and Quantitative Real Time Polymerase Chain Reaction (qPCR. Biofilms of L. monocytogenes were obtained in stainless steel coupons immersed in Brain Heart Infusion Broth, under agitation at 37 °C for 24 h. The methods selected for this study were based on plate count, microscopic count with the aid of viability dyes (CTC-DAPI, and qPCR. Results of culture method showed that peroxyacetic acid was efficient to kill sessile L. monocytogenes populations, while sodium hypochlorite was only partially effective to kill attached L. monocytogenes (p < 0.05. When, viability dyes (CTC/DAPI combined with fluorescence microscopy and qPCR were used and lower counts were found after treatments (p < 0.05. Selective quantification of viable cells of L. monocytogenes by qPCR using EMA revelead that the pre-treatment with EMA was not appropriate since it also inhibited amplification of DNA from live cells by ca. 2 log. Thus, the use of CTC counts was the best method to count viable cells in biofilms.

  15. Perfusion Computed Tomography for the Assessment of Myocardial Viability — a Case Series

    Directory of Open Access Journals (Sweden)

    Morariu Mirabela

    2016-06-01

    Full Text Available Myocardial viability plays an important role in preventing the development of left ventricular remodeling following an acute myocardial infarction. A preserved viability in the infarcted area has been demonstrated to be associated with a lower amplitude of the remodeling process, while the extent of the non-viable myocardium is directly correlated with the amplitude of the remodeling process. A number of methods are currently in use for the quantification of the viable myocardium, and some of them are based on the estimation of myocardial perfusion during pharmacologic stress. 64-slice Multi-detector Computed Tomography (MDCT during vasodilator stress test, associated with CT Coronary Angiography (CCTA has a high diagnostic accuracy in evaluating myocardial perfusion. In this article, we present a sequence of 3 clinical cases that presented with symptoms of myocardial ischemia, who underwent 64-slice MDCT imaging at rest and during adenosine stress test, in order to assess the extent of the hypoperfused myocardial areas. Coronary artery anatomy and the Coronary Calcium Score was assessed for all 3 patients by performing CT Coronary Angiography. The combination of CT Angiography and adenosine stress CT myocardial perfusion imaging can accurately detect atherosclerosic lesions that cause perfusion abnormalities, compared with the combination of invasive angiography and single-photon emission computed tomography (SPECT.

  16. Challenges in Learning for Company‘s Financial Viability Assessment and Management

    Directory of Open Access Journals (Sweden)

    Nadezda Koleda

    2015-06-01

    Full Text Available Many entrepreneurs in Latvia are limited in choice of tools for the improvement of their financial literacy. Therefore the training course “Financial literacy of entrepreneurs in the field of financial viability management” was elaborated and tested with the aim to improve the course and reveal the challenges facing different categories of learners. The research was formative as it combined the promotion of learners and trainers’ competencies and behaviours with the research process throughout all the five training cycles. The research methods applied were chosen based on the inner logics and aims of the teaching and learning processes: videorecording of learners’ activities and analysis of the videomaterials; analysis of the learners’ work and reflections. Out of the identified sixteen challenges faced by learners, three were especially crucial and caused further difficulties in coping with different tasks on assessment and management of company’s financial viability. It was concluded that there may be three categories of learners whose training should be organised in different modes taking into account the level of their preparedness in accountancy, tax accounting and financial analysis.

  17. E-cigarettes: voltage- and concentration-dependent loss in human lung adenocarcinoma viability.

    Science.gov (United States)

    Otręba, Michał; Kośmider, Leon; Knysak, Jakub; Warncke, Jared D; Sobczak, Andrzej

    2018-04-17

    E-cigarettes are used by millions of people despite the fact that the harmful effect of aerosol emitted from these products to the human organism is still not clear. In this paper, toxicity of vapor generated using different solutions and battery output voltage on A549 cells viability is presented. The obtained EC 50 values for commercially available propylene glycol/glycerol solution 1:1 e-liquids based on 3.2 V (0.127%), 4.0 V (0.112%) and 4.8 V (0.038%) were about 1.5-4.5 times higher than in tobacco smoke (0.0086%). Furthermore, it was shown that the increase of battery output voltage decreased A549 cell viability. In addition, commercially available extracts were more cytotoxic than laboratory made extracts. Owing to the expansiveness of e-cigarettes, it is very important to estimate their impact on public health. Our results not only confirm less cytotoxicity of e-liquid aerosol than cigarette smoke, but also demonstrate that solutions used in e-liquids and, for the first time, battery output voltage have a significant impact on cytotoxicity of e-cigarette vapor. Thus, the results of this study are very important for the current and future legal regulations on e-cigarettes. Copyright © 2018 John Wiley & Sons, Ltd.

  18. Edaravone enhances the viability of ischemia/reperfusion flaps.

    Science.gov (United States)

    Zhang, Dong-Yi; Kang, Shen-Song; Zhang, Zheng-Wen; Wu, Rui

    2017-02-01

    The purpose of the experiment was to study the efficacy of edaravone in enhancing flap viability after ischemia/reperfusion (IR) and its mechanism. Forty-eight adult male SD rats were randomly divided into 3 groups: control group (n=16), IR group (n=16), and edaravone-treated IR group (n=16). An island flap at left lower abdomen (6.0 cm×3.0 cm in size), fed by the superficial epigastric artery and vein, was created in each rat of all the three groups. The arterial blood flow of flaps in IR group and edaravone-treated IR group was blocked for 10 h, and then the blood perfusion was restored. From 15 min before reperfusion, rats in the edaravone-treated IR group were intraperitoneally injected with edaravone (10 mg/kg), once every 12 h, for 3 days. Rats in the IR group and control group were intraperitoneally injected with saline, with the same method and frequency as the rats in the edaravone-treated IR group. In IR group and edaravone-treated IR group, samples of flaps were harvested after reperfusion of the flaps for 24 h. In the control group, samples of flaps were harvested 34 h after creation of the flaps. The content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) were determined, and changes in organizational structure and infiltration of inflammatory cells were observed by hematoxylin-eosin (HE) staining, apoptotic cells of vascular wall were marked by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, and the apoptotic rate of cells in vascular wall was calculated. The ultrastructural changes of vascular endothelial cells were observed by transmission electron microscopy (TEM). Seven days after the operation, we calculated the flap viability of each group, and marked vessels of flaps by immunohistochemical staining for calculating the average number of subcutaneous vessels. The results showed that the content of MDA, the number of multicore inflammatory cells and apoptotic rate of cells in vascular wall

  19. Technical viability and development needs for waste forms and facilities

    Energy Technology Data Exchange (ETDEWEB)

    Pegg, I.; Gould, T.

    1996-05-01

    The objective of this breakout session was to provide a forum to discuss technical issues relating to plutonium-bearing waste forms and their disposal facilities. Specific topics for discussion included the technical viability and development needs associated with the waste forms and/or disposal facilities. The expected end result of the session was an in-depth (so far as the limited time would allow) discussion of key issues by the session participants. The session chairs expressed allowance for, and encouragement of, alternative points of view, as well as encouragement for discussion of any relevant topics not addressed in the paper presentations. It was not the intent of this session to recommend or advocate any one technology over another.

  20. Cosmological viability of theories with massive spin-2 fields

    Energy Technology Data Exchange (ETDEWEB)

    Koennig, Frank

    2017-03-30

    Theories of spin-2 fields take on a particular role in modern physics. They do not only describe the mediation of gravity, the only theory of fundamental interactions of which no quantum field theoretical description exists, it furthermore was thought that they necessarily predict massless gauge bosons. Just recently, a consistent theory of a massive graviton was constructed and, subsequently, generalized to a bimetric theory of two interacting spin-2 fields. This thesis studies both the viability and consequences at cosmological scales in massive gravity as well as bimetric theories. We show that all consistent models that are free of gradient and ghost instabilities behave like the cosmological standard model, LCDM. In addition, we construct a new theory of massive gravity which is stable at both classical background and quantum level, even though it suffers from the Boulware-Deser ghost.

  1. Microbial viability in preparations packaged for single use.

    Science.gov (United States)

    Obayashi, Akiko; Oie, Shigeharu; Kamiya, Akira

    2003-05-01

    We evaluated microbial viability in preparations packaged for single use only which mandate that residual solution be discarded such as albumin and globulin preparations as blood products, preparations containing albumin (such as urokinase and interferon), fat emulsions, and a preparation containing fat emulsions (propofol). In most preparations, Serratia marcescens and Burkholderia cepacia proliferated rapidly at 30 degrees C. However, in globulin preparations containing 1-2.25% glycine to prevent protein degradation (Gamma-Venin P, Venilon-I, Globulin Injection, and Ahlbulin), no growth of S. marcescens and B. cepacia was detected over 24 h at 30 degrees C. For globulin preparations containing 1-2.25% glycine, the injunction to "Discard residual solution after the package has been used" in the package inserts can be revised to "It is possible to use residual solution within 24 h after the package has been used with storage in a cool place."

  2. Modelling the viability of heat recovery from combined sewers.

    Science.gov (United States)

    Abdel-Aal, M; Smits, R; Mohamed, M; De Gussem, K; Schellart, A; Tait, S

    2014-01-01

    Modelling of wastewater temperatures along a sewer pipe using energy balance equations and assuming steady-state conditions was achieved. Modelling error was calculated, by comparing the predicted temperature drop to measured ones in three combined sewers, and was found to have an overall root mean squared error of 0.37 K. Downstream measured wastewater temperature was plotted against modelled values; their line gradients were found to be within the range of 0.9995-1.0012. The ultimate aim of the modelling is to assess the viability of recovering heat from sewer pipes. This is done by evaluating an appropriate location for a heat exchanger within a sewer network that can recover heat without impacting negatively on the downstream wastewater treatment plant (WWTP). Long sewers may prove to be more viable for heat recovery, as heat lost can be reclaimed before wastewater reaching the WWTP.

  3. Viability report for the ByWater Lakes project

    Energy Technology Data Exchange (ETDEWEB)

    Lowry, Thomas Stephen [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Klise, Geoffrey Taylor [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Passell, Howard David [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Peplinski, William J. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2013-10-01

    This report presents the results from the hydrological, ecological, and renewable energy assessments conducted by Sandia National Laboratories at the ByWater Lakes site in Espanola, New Mexico for ByWater Recreation LLC and Avanyu Energy Services through the New Mexico small business assistance (NMSBA) program. Sandia's role was to assess the viability and provide perspective for enhancing the site to take advantage of renewable energy resources, improve and sustain the natural systems, develop a profitable operation, and provide an asset for the local community. Integral to this work was the identification the pertinent data and data gaps as well as making general observations about the potential issues and concerns that may arise from further developing the site. This report is informational only with no consideration with regards to the business feasibility of the various options that ByWater and Avanyu may be pursuing.

  4. Phenomenological viability of orbifold models with three Higgs families

    International Nuclear Information System (INIS)

    Escudero, Nicolas; Munoz, Carlos; Teixeira, Ana M.

    2006-01-01

    We discuss the phenomenological viability of string multi-Higgs doublet models, namely a scenario of heterotic Z 3 orbifolds with two Wilson lines, which naturally predicts three supersymmetric families of matter and Higgs fields. We study the orbifold parameter space, and discuss the compatibility of the predicted Yukawa couplings with current experimental data. We address the implications of tree-level flavour changing neutral processes in constraining the Higgs sector of the model, finding that viable scenarios can be obtained for a reasonably light Higgs spectrum. We also take into account the tree-level contributions to indirect CP violation, showing that the experimental value of ε K can be accommodated in the present framework

  5. Relationship between humidity and influenza A viability in droplets and implications for influenza's seasonality.

    Directory of Open Access Journals (Sweden)

    Wan Yang

    Full Text Available Humidity has been associated with influenza's seasonality, but the mechanisms underlying the relationship remain unclear. There is no consistent explanation for influenza's transmission patterns that applies to both temperate and tropical regions. This study aimed to determine the relationship between ambient humidity and viability of the influenza A virus (IAV during transmission between hosts and to explain the mechanisms underlying it. We measured the viability of IAV in droplets consisting of various model media, chosen to isolate effects of salts and proteins found in respiratory fluid, and in human mucus, at relative humidities (RH ranging from 17% to 100%. In all media and mucus, viability was highest when RH was either close to 100% or below ∼50%. When RH decreased from 84% to 50%, the relationship between viability and RH depended on droplet composition: viability decreased in saline solutions, did not change significantly in solutions supplemented with proteins, and increased dramatically in mucus. Additionally, viral decay increased linearly with salt concentration in saline solutions but not when they were supplemented with proteins. There appear to be three regimes of IAV viability in droplets, defined by humidity: physiological conditions (∼100% RH with high viability, concentrated conditions (50% to near 100% RH with lower viability depending on the composition of media, and dry conditions (<50% RH with high viability. This paradigm could help resolve conflicting findings in the literature on the relationship between IAV viability in aerosols and humidity, and results in human mucus could help explain influenza's seasonality in different regions.

  6. Dendritic Cells Promote Pancreatic Viability in Mice with Acute Pancreatitis

    Science.gov (United States)

    Bedrosian, Andrea S.; Nguyen, Andrew H.; Hackman, Michael; Connolly, Michael K.; Malhotra, Ashim; Ibrahim, Junaid; Cieza-Rubio, Napoleon E.; Henning, Justin R.; Barilla, Rocky; Rehman, Adeel; Pachter, H. Leon; Medina-Zea, Marco V.; Cohen, Steven M.; Frey, Alan B.; Acehan, Devrim; Miller, George

    2011-01-01

    Background & Aims Acute pancreatitis increases morbidity and mortality from organ necrosis by mechanisms that are incompletely understood. Dendritic cells (DCs) can promote or suppress inflammation, depending on their subtype and context. We investigated the roles of DC in development of acute pancreatitis. Methods Acute pancreatitis was induced in CD11c.DTR mice using caerulein or L-arginine; DCs were depleted by administration of diphtheria toxin. Survival was analyzed using Kaplan-Meier analysis. Results Numbers of MHC II+CD11c+DC increased 100-fold in pancreas of mice with acute pancreatitis, to account for nearly 15% of intra-pancreatic leukocytes. Intra-pancreatic DC acquired an immune phenotype in mice with acute pancreatitis; they expressed higher levels of MHC II and CD86 and increased production of interleukin-6, membrane cofactor protein (MCP)-1, and tumor necrosis factor (TNF)-α. However, rather than inducing an organ-destructive inflammatory process, DC were required for pancreatic viability; the exocrine pancreas died in mice that were depleted of DC and challenged with caerulein or L-arginine. All mice with pancreatitis that were depleted of DC died from acinar cell death within 4 days. Depletion of DC from mice with pancreatitis resulted in neutrophil infiltration and increased levels of systemic markers of inflammation. However, the organ necrosis associated with depletion of DC did not require infiltrating neutrophils, activation of NF-κB, or signaling by mitogen-activated protein kinase or TNF-α. Conclusions DC are required for pancreatic viability in mice with acute pancreatitis and might protect organs against cell stress. PMID:21801698

  7. High speed measurement of corn seed viability using hyperspectral imaging

    Science.gov (United States)

    Ambrose, Ashabahebwa; Kandpal, Lalit Mohan; Kim, Moon S.; Lee, Wang-Hee; Cho, Byoung-Kwan

    2016-03-01

    Corn is one of the most cultivated crops all over world as food for humans as well as animals. Optimized agronomic practices and improved technological interventions during planting, harvesting and post-harvest handling are critical to improving the quantity and quality of corn production. Seed germination and vigor are the primary determinants of high yield notwithstanding any other factors that may play during the growth period. Seed viability may be lost during storage due to unfavorable conditions e.g. moisture content and temperatures, or physical damage during mechanical processing e.g. shelling, or over heating during drying. It is therefore vital for seed companies and farmers to test and ascertain seed viability to avoid losses of any kind. This study aimed at investigating the possibility of using hyperspectral imaging (HSI) technique to discriminate viable and nonviable corn seeds. A group of corn samples were heat treated by using microwave process while a group of seeds were kept as control group (untreated). The hyperspectral images of corn seeds of both groups were captured between 400 and 2500 nm wave range. Partial least squares discriminant analysis (PLS-DA) was built for the classification of aged (heat treated) and normal (untreated) corn seeds. The model showed highest classification accuracy of 97.6% (calibration) and 95.6% (prediction) in the SWIR region of the HSI. Furthermore, the PLS-DA and binary images were capable to provide the visual information of treated and untreated corn seeds. The overall results suggest that HSI technique is accurate for classification of viable and non-viable seeds with non-destructive manner.

  8. Economic and financial viability of small-scale dairy systems in central Mexico: economic scenario 2010-2018.

    Science.gov (United States)

    Posadas-Domínguez, R R; Callejas-Juárez, N; Arriaga-Jordán, C M; Martínez-Castañeda, F E

    2016-12-01

    A simulation Monte Carlo model was used to assess the economic and financial viability of 130 small-scale dairy farms in central Mexico, through a Representative Small-Scale Dairy Farm. Net yields were calculated for a 9-year planning horizon by means of simulated values for the distribution of input and product prices taking 2010 as base year and considering four scenarios which were compared against the scenario of actual production. The other scenarios were (1) total hiring in of needed labour; (2) external purchase of 100 % of inputs and (3) withdrawal of subsidies to production. A stochastic modelling approach was followed to determine the scenario with the highest economic and financial viability. Results show a viable economic and financial situation for the real production scenario, as well as the scenarios for total hiring of labour and of withdrawal of subsidies, but the scenario when 100 % of feed inputs for the herd are bought-in was not viable.

  9. Biomass viability: An experimental study and the development of an empirical mathematical model for submerged membrane bioreactor.

    Science.gov (United States)

    Zuthi, M F R; Ngo, H H; Guo, W S; Nghiem, L D; Hai, F I; Xia, S Q; Zhang, Z Q; Li, J X

    2015-08-01

    This study investigates the influence of key biomass parameters on specific oxygen uptake rate (SOUR) in a sponge submerged membrane bioreactor (SSMBR) to develop mathematical models of biomass viability. Extra-cellular polymeric substances (EPS) were considered as a lumped parameter of bound EPS (bEPS) and soluble microbial products (SMP). Statistical analyses of experimental results indicate that the bEPS, SMP, mixed liquor suspended solids and volatile suspended solids (MLSS and MLVSS) have functional relationships with SOUR and their relative influence on SOUR was in the order of EPS>bEPS>SMP>MLVSS/MLSS. Based on correlations among biomass parameters and SOUR, two independent empirical models of biomass viability were developed. The models were validated using results of the SSMBR. However, further validation of the models for different operating conditions is suggested. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Determination of viability of preserved skin in low temperature

    International Nuclear Information System (INIS)

    Yang Hong Chang; Hao Zheng Ming; Zao Xiao Chun

    1999-01-01

    The skin from fresh human cadavers was stored in 4-18 degree C refrigerator. Before it was grafted for treatment of burn patients, it was quickly put into 40 degree C water and bring back to a former condition. The survival rate of skin was related with time and temperature of store. We used oxygen consumption to observe the change of viability of preserved skin. Oxygen consumption of skin was observed with apparatus made in the 304th Hospital of Peoples Liberation Army. The operating temperature was 5 - 45 degree C. Determination range was 0 - 199 mm Hg, resolving power of digital display was I mm Hg, instrumental error < 0.5 s'. Fresh human cadavers skin was made into 0.3 - 0.4 mm thick piece. Cleaned with NaCl 0.9% for three time. Then it was kept in neomycin solution for fifteen minutes. Then cut into 0.5 x 0.5 cm slices and stored in neomycin (2mg/ml). The skin was stored in 4 degree C refrigerator for five different periods (1, 2, 3, 5 and 7 days). Then the Oxygen consumption was determined immediately. The oxygen consumption was also determined before and after it was stored for 24 hours. After the skin was stored in 4 and -18 degree C for 24 hours the oxygen consumption was determined immediately. The prepared skin, which was stored in ordinary refrigerator, was useful and simple. The preserved skin was grafted onto the bum patient and survival rate was high and in short time. But the result showed the viability of preserved skin reduced with time. The result showed that the oxygen consumption of skin, which was stored at 4 degree C, on the fifth day was 62.23% and on day 7 was 30.5%. The study showed that the preserved skin which was stored at 4 degree C for five days was better while the vitality of skin evidently reduced after seven days and the survival rate was low. The oxygen consumption of preserved skin that was stored in -18 degree C refrigerator for 24 hours was 100%. But in 4 degree C refrigerator it was 89.1%. The result showed that the

  11. Identifying genetic variants that affect viability in large cohorts.

    Directory of Open Access Journals (Sweden)

    Hakhamanesh Mostafavi

    2017-09-01

    Full Text Available A number of open questions in human evolutionary genetics would become tractable if we were able to directly measure evolutionary fitness. As a step towards this goal, we developed a method to examine whether individual genetic variants, or sets of genetic variants, currently influence viability. The approach consists in testing whether the frequency of an allele varies across ages, accounting for variation in ancestry. We applied it to the Genetic Epidemiology Research on Adult Health and Aging (GERA cohort and to the parents of participants in the UK Biobank. Across the genome, we found only a few common variants with large effects on age-specific mortality: tagging the APOE ε4 allele and near CHRNA3. These results suggest that when large, even late-onset effects are kept at low frequency by purifying selection. Testing viability effects of sets of genetic variants that jointly influence 1 of 42 traits, we detected a number of strong signals. In participants of the UK Biobank of British ancestry, we found that variants that delay puberty timing are associated with a longer parental life span (P~6.2 × 10-6 for fathers and P~2.0 × 10-3 for mothers, consistent with epidemiological studies. Similarly, variants associated with later age at first birth are associated with a longer maternal life span (P~1.4 × 10-3. Signals are also observed for variants influencing cholesterol levels, risk of coronary artery disease (CAD, body mass index, as well as risk of asthma. These signals exhibit consistent effects in the GERA cohort and among participants of the UK Biobank of non-British ancestry. We also found marked differences between males and females, most notably at the CHRNA3 locus, and variants associated with risk of CAD and cholesterol levels. Beyond our findings, the analysis serves as a proof of principle for how upcoming biomedical data sets can be used to learn about selection effects in contemporary humans.

  12. Monitoring the viability of citrus rootstocks seeds stored under refrigeration

    Directory of Open Access Journals (Sweden)

    Sérgio Alves de Carvalho

    2013-03-01

    Full Text Available The citrus nursery tree is produced through the bud grafting process, in which rootstock is usually grown from seed germination. The objective of this research was to evaluate, in two dissimilar environmental conditions, the viability and polyembryony expression of five citrus rootstocks seeds stored in different periods under refrigeration. The rootstock varieties evaluated were: Rangpur lime (Citrus limonia Osb. cv. Limeira, Trifoliate orange (Poncirus trifoliata Raf. cv. Limeira, Citrumelo (P. trifoliata x C. paradisi Macf. cv. Swingle, Sunki mandarin (C. sunki Hort. ex Tanaka and Volkamer lemon (C. volkameriana Ten. & Pasq. cv. Catania 2. The experimental design was the randomized blocks in a 11 x 5 x 2 factorial scheme, evaluating from time zero to the tenth month of storage, the five varieties of rootstock in two environments: germination and growth B.O.D type chamber (Biological Oxygen Demand - Eletrolab Brand Model FC 122 at 25 °C; and greenhouse seedbed with partial temperature control (22 °C to 36 °C and humidity control (75-85%. The plot had 24 seeds in four replicates, using trays with substrate in greenhouse and Petri dishes with filter paper in B.O.D. chamber. The seed germination rate and polyembryony expression were evaluated monthly. It was concluded that Trifoliate and Citrumelo Swingle seeds can be stored for up to seven months, while Volkamer lemon, Rangpur lime and Sunki seeds can be stored for up to ten months. The polyembryony expression rate was slightly higher when measured in greenhouse than in B.O.D. chamber and remained stable in both environments until the seventh month, from which dropped sharply. Citrumelo Swingle seeds expressed the highest polyembryony rate (18.8%, followed by Rangpur lime and Volkamer lemon (average value of 13.7%, Sunki (9.4% and Trifoliate (3.2%. Despite some differences among varieties, the viability of rootstock stored seeds can be monitored either in the greenhouse or in B

  13. Population Viability Analysis of feral raccoon dog (Nyctereutes procyonoides) in Denmark

    DEFF Research Database (Denmark)

    Rømer, Anna Elisabeth; Nørgaard, Louise Solveig; Mikkelsen, Dorthe Malene Götz

    2015-01-01

    that more 2 efficient and intensive actions are needed to reach the goal of the DAP, aiming at eradicating the breeding population of raccoon dogs in Denmark within 2015. Simulations suggested that around 950 individuals should be culled a year from 2012 to 2015. Sensitivity analysis that was performed......To assess the effects of actions implemented by the Danish Action Plan (DAP) for eradication of the raccoon dog, the population dynamics of the raccoon dog in Denmark was simulated. A population viability analysis (PVA) was generated with the stochastic simulation program, VORTEX, based...... and reach an assessed carrying capacity of 30 000 individuals with no intervention within 10 years. Simulations of the current culling strategy showed that the raccoon dog in Denmark would reach the carrying capacity with only a few years' delay compared to simulations with no intervention. This indicates...

  14. Technic and economic viability study on exploitation of lunar 3He resource

    International Nuclear Information System (INIS)

    Deng Baiquan

    1995-01-01

    From the energetics point of view, the technic and economic viability study on exploitation of lunar 3 He for fuelling the fusion reactor burning D- 3 He has been carried out. This study is divided into the following sections: analysis of solar wind parameters and estimation of potential quantity 3 He in the lunar regolith, the cost evaluation of mining He of lunar soil; the energy cost calculation of He extraction by vacuum heating degassing during lunar day, the cost calculation of cryogenic isotopic separation 3 He/ 4 He during the lunar night, the energy cost for earth/moon transportation of liquid 3 He, the energy payback calculation of fusion power burning 3 He based lunar source, and finally the comparison of the energy multiplication with that for 235 U production of nuclear fuel and for coal mining. The comparisons of cost of electricity between D- 3 He and D-T fuel cycle for different reactor types have been discussed

  15. Growth and viability of Aedes albopictus cell line in vitro after cesium-137 gamma irradiation

    International Nuclear Information System (INIS)

    Blakely, E.A.

    1975-01-01

    The radiosensitivity of the cultured mosquito cell line Aedes albopictus (Skuse) was investigated. Population growth was followed by total cell counts and by viable cell counts on aliquots of cultures exposed to various doses of gamma radiation during exponential growth. Viable cell determinations were based on the cellular exclusion of the dye, alcian blue, in a procedure adapted to the insect cells in culture. Viability determinations in the irradiated exponential cultures indicated that initially there was some increase in the gestation, suggesting that gonadal steroids may have unusual effects on uterine physiology and biochemistry in this species. Consequently, studies were undertaken to elucidate some of the basic responses of hamster uteri to estradiol benzoate and progesterone under conditions of protein malnutrition, actinomycin D administration and corticosterone injection. Furthermore, the effects of gonadal steroids on uteri of pregnant ovariectomized hamsters were studied

  16. Viability analysis of EMBRAPA's swine manure treatment system

    Energy Technology Data Exchange (ETDEWEB)

    Miele, Marcelo; Kunz, Airton; Seganfredo, Milton Antonio; Steinmetz, Ricardo [EMBRAPA Suinos e Aves, Concordia, SC (Brazil)], E-mail: mmiele@cnpsa.embrapa.br; Bortoli, Marcelo [Universidade Federal de Santa Catarina (UFSC), Florianopolis, SC (Brazil)

    2008-07-01

    The swine manure treatment is based on several technologies, such as pond systems, composting, biodigesters, and compact process like the EMBRAPA's Swine Manure Treatment System. The last one can reach high quality effluent with reduced pollution, although it demands higher investment and costs. For this reason it is necessary to manage revenues with byproducts and carbon credit markets. The aim of this study was to present a viability analysis of EMBRAPA's swine manure treatment system considering its potential revenues facing higher costs. Using market prices and measured data from a prototype running in south Brazil, the study calculated costs, revenues, profit, net present value and internal rate return. It also estimated these indicators assuming scale economies. The results showed that the investment can pay its costs by its insertion in the carbon credit market and other revenues like the energy substitution. It also showed the importance to reach scale economies. (author)

  17. Viability and resilience of complex systems concepts, methods and case studies from ecology and society

    CERN Document Server

    Deffuant, Guillaume

    2011-01-01

    One common characteristic of a complex system is its ability to withstand major disturbances and the capacity to rebuild itself. Understanding how such systems demonstrate resilience by absorbing or recovering from major external perturbations requires both quantitative foundations and a multidisciplinary view of the topic. This book demonstrates how new methods can be used to identify the actions favouring the recovery from perturbations on a variety of examples including the dynamics of bacterial biofilms, grassland savannahs, language competition and Internet social networking sites. The reader is taken through an introduction to the idea of resilience and viability and shown the mathematical basis of the techniques used to analyse systems. The idea of individual or agent-based modelling of complex systems is introduced and related to analytically tractable approximations of such models. A set of case studies illustrates the use of the techniques in real applications, and the final section describes how on...

  18. Impact of different spray-drying conditions on the viability of wine Saccharomyces cerevisiae strains.

    Science.gov (United States)

    Aponte, Maria; Troianiello, Gabriele Danilo; Di Capua, Marika; Romano, Raffaele; Blaiotta, Giuseppe

    2016-01-01

    Spray-drying (SD) is widely considered a suitable method to preserve microorganisms, but data regarding yeasts are still scanty. In this study, the effect of growing media, process variables and carriers over viability of a wild wine Saccharomyces (S.) cerevisiae LM52 was evaluated. For biomass production, the strain was grown (batch and fed-batch fermentation) in a synthetic, as well as in a beet sugar molasses based-medium. Drying of cells resuspended in several combinations of soluble starch and maltose was performed at different inlet and outlet temperatures. Under the best conditions-suspension in soluble starch plus maltose couplet to inlet and outlet temperatures of 110 and 55 °C, respectively-the loss of viability of S. cerevisiae LM52 was 0.8 ± 0.1 and 0.5 ± 0.2 Log c.f.u. g(-1) for synthetic and molasses-based medium, respectively. Similar results were obtained when S. cerevisiae strains Zymoflore F15 and EC1118, isolated from commercial active dry yeast (ADY), were tested. Moreover, powders retained a high vitality and showed good fermentation performances up to 6 month of storage, at both 4 and -20 °C. Finally, fermentation performances of different kinds of dried formulates (SD and ADY) compared with fresh cultures did not show significant differences. The procedure proposed allowed a small-scale production of yeast in continuous operation with relatively simple equipment, and may thus represent a rapid response-on-demand for the production of autochthonous yeasts for local wine-making.

  19. Using Bayesian Population Viability Analysis to Define Relevant Conservation Objectives.

    Directory of Open Access Journals (Sweden)

    Adam W Green

    Full Text Available Adaptive management provides a useful framework for managing natural resources in the face of uncertainty. An important component of adaptive management is identifying clear, measurable conservation objectives that reflect the desired outcomes of stakeholders. A common objective is to have a sustainable population, or metapopulation, but it can be difficult to quantify a threshold above which such a population is likely to persist. We performed a Bayesian metapopulation viability analysis (BMPVA using a dynamic occupancy model to quantify the characteristics of two wood frog (Lithobates sylvatica metapopulations resulting in sustainable populations, and we demonstrate how the results could be used to define meaningful objectives that serve as the basis of adaptive management. We explored scenarios involving metapopulations with different numbers of patches (pools using estimates of breeding occurrence and successful metamorphosis from two study areas to estimate the probability of quasi-extinction and calculate the proportion of vernal pools producing metamorphs. Our results suggest that ≥50 pools are required to ensure long-term persistence with approximately 16% of pools producing metamorphs in stable metapopulations. We demonstrate one way to incorporate the BMPVA results into a utility function that balances the trade-offs between ecological and financial objectives, which can be used in an adaptive management framework to make optimal, transparent decisions. Our approach provides a framework for using a standard method (i.e., PVA and available information to inform a formal decision process to determine optimal and timely management policies.

  20. Viability of microencapsulated Lactobacillus casei in synbiotic mayonnaise

    Directory of Open Access Journals (Sweden)

    Lieu, M.D.

    2017-07-01

    Full Text Available In this study, whey protein, maltodextrin and GOS (Galacto-oligosaccharides used as microencapsulating agents to protect Lactobacillus casei during spray-drying and mayonnaise storage. The morphology of microcapsules, pH charges, the survival rate during mayonnaise storage as well as survival in simulated gastric fluid (SGF and intestinal fluid (SIF was tested in this study. The results indicated that whey protein showed a protective effect better than maltodextrin during spray-drying. The particles showed spherical shape and typical concavity of all samples and encapsulating agents were not affected by the size and surface structure of particles. The pH charges were not significantly different in all mayonnaise samples in this test. The viability of free cell L. casei after 6 weeks storage was significant decrease about 4 log CFU/g compared to 1.55 to 3.27 log CFU/g in the mayonnaise samples containing microcapsules in which maltodextrin showed the lowest of L. casei survival rate. In SGF and SIF conditions, maltodextrin act as prebiotic sufficiently which do not need adding GOS. The combination of whey protein and maltodextrin in which maltodextrin plays a role as supporting agents for the spray-drying process as well as prebiotic potential, while whey protein with high buffer property which enhancing the survival rate of L. casie in low pH.

  1. A Method for Quantitative Determination of Biofilm Viability

    Directory of Open Access Journals (Sweden)

    Maria Strømme

    2012-06-01

    Full Text Available In this study we present a scheme for quantitative determination of biofilm viability offering significant improvement over existing methods with metabolic assays. Existing metabolic assays for quantifying viable bacteria in biofilms usually utilize calibration curves derived from planktonic bacteria, which can introduce large errors due to significant differences in the metabolic and/or growth rates of biofilm bacteria in the assay media compared to their planktonic counterparts. In the presented method we derive the specific growth rate of Streptococcus mutans bacteria biofilm from a series of metabolic assays using the pH indicator phenol red, and show that this information could be used to more accurately quantify the relative number of viable bacteria in a biofilm. We found that the specific growth rate of S. mutans in biofilm mode of growth was 0.70 h−1, compared to 1.09 h−1 in planktonic growth. This method should be applicable to other bacteria types, as well as other metabolic assays, and, for example, to quantify the effect of antibacterial treatments or the performance of bactericidal implant surfaces.

  2. Chemicals and lemon essential oil effect on Alicyclobacillus acidoterrestris viability

    Directory of Open Access Journals (Sweden)

    Maria Cristina Maldonado

    2013-12-01

    Full Text Available Alicyclobacillus acidoterrestris is considered to be one of the important target microorganisms in the quality control of acidic canned foods. There is an urgent need to develop a suitable method for inhibiting or controlling the germination and outgrowth of A.acidoterrestris in acidic drinks. The aim of this work was to evaluate the chemicals used in the lemon industry (sodium benzoate, potassium sorbate, and lemon essential oil as a natural compound, against a strain of A.acidoterrestris in MEB medium and in lemon juice concentrate. The results pointed out that sodium benzoate (500-1000-2000 ppm and lemon essential oil (0.08- 0.12- 0.16% completely inhibited the germination of A. acidoterrestris spores in MEB medium and LJC for 11 days. Potassium sorbate (600-1200 ppm was more effective to inhibit the growth of the microbial target in lemon juice than in MEB medium. The effect of sodium benzoate, potassium sorbate and essential oil was sporostatic in MEB and LJC as they did not affect spore viability.

  3. Open pit coal exploitation viability. Margarita mine. Case of study

    International Nuclear Information System (INIS)

    Veloza, Julia; Molina, Jorge; Mejia, Humberto

    2006-01-01

    This paper provides an analysis of financial viability, planning and design for the new coal open pit exploitation for La Margarita mine, with coal-resources estimated on 440.139,7 ton. Dimension, design and economic evaluation were possible by three exploitation methods: (multiple bench, open cast contour, and terraces). Net present values (NVP) were calculated: $c 817,5; $c 518,5 and $c 645,2 respectively for each method (given in million current Colombian pesos $. $c 2380 are equivalent to $us 1) and rate of return (ROR) 78,33%; 34,0% and 38,62% respectively for each method. These indicators served as a parameter to choose the multiple bench method, which should be recalculated because it was necessary to work jointly with two pits and making feasible the project. in addition a general environmental evaluation was done, which is vital for the exploitation. Important impacts on the flower, animals, air, water were found, and measures of control, prevention and mitigation were stated. it is expected that this paper can be useful as a technical-economical support for the development of the open pit exploitation in the margarita mine

  4. Assaying Cellular Viability Using the Neutral Red Uptake Assay.

    Science.gov (United States)

    Ates, Gamze; Vanhaecke, Tamara; Rogiers, Vera; Rodrigues, Robim M

    2017-01-01

    The neutral red uptake assay is a cell viability assay that allows in vitro quantification of xenobiotic-induced cytotoxicity. The assay relies on the ability of living cells to incorporate and bind neutral red, a weak cationic dye, in lysosomes. As such, cytotoxicity is expressed as a concentration-dependent reduction of the uptake of neutral red after exposure to the xenobiotic under investigation. The neutral red uptake assay is mainly used for hazard assessment in in vitro toxicology applications. This method has also been introduced in regulatory recommendations as part of 3T3-NRU-phototoxicity-assay, which was regulatory accepted in all EU member states in 2000 and in the OECD member states in 2004 as a test guideline (TG 432). The present protocol describes the neutral red uptake assay using the human hepatoma cell line HepG2, which is often employed as an alternative in vitro model for human hepatocytes. As an example, the cytotoxicity of acetaminophen and acetyl salicylic acid is assessed.

  5. A note on the viability of Gauss-Bonnet cosmology

    International Nuclear Information System (INIS)

    Chingangbam, R.; Sami, M.; Tretyakov, P.V.; Toporensky, A.V.

    2008-01-01

    In this Letter, we analyze the viability of a vacuum Gauss-Bonnet cosmology by examining the dynamics of the homogeneous and anisotropic background in 4+1 dimensions. The trajectories of the system either originate from the standard singularity or from non-standard type, the later is characterized by the divergence of time derivative of the Hubble parameters for its finite value. At the onset, the system should relax to Einstein phase at late times as the effect of Gauss-Bonnet term becomes negligible in the low energy regime. However, we find that most of the trajectories emerging from the standard big-bang singularity lead to future re-collapse whereas the system beginning its evolution from the non-standard singularity enters the Kasner regime at late times. This leads to the conclusion that the measure of trajectories giving rise to a smooth evolution from a standard singularity to the Einstein phase is negligibly small for generic initial conditions

  6. Bone graft viability evaluated by three phase bone scan

    International Nuclear Information System (INIS)

    Ljiljana Jaukovic Rajko Spaic; Marijan Novakovic; Srbislav Stosic

    2004-01-01

    Bone defects resulting war injury can be replaced by microvascular bone grafts from fibula. Aim: The aim of this study was to assess the value of three phase (3P) bone scintigraphy in the early detection of the bone graft complications. Method: 3P bone scans were performed in four patients (two after mandible reconstruction with micro vascular fibular bone grafts, one after fibular transplantation for ulnar and one with humeral reconstruction). First dynamic phase scan was performed immediately after iv injection of 740 MBq Tc- 99m DPD, acquiring 15 two seconds duration frames. Second, early static scan was performed during next 300 seconds, and third, delayed scan three hours later. All scans were obtained under the bone graft region. The scans were evaluated using ROI under graft region and the corresponding contra lateral area. Blood flow in graft region was determined using first phase scan, and tracer uptake in the same region was determined using second and third phase scans. Results: in all patients blood flow in graft region was particularly normal. Tracer uptake in one of two patients with mandible reconstruction was diffusely increased in graft, strongly suggesting infection; In the other patient delayed scan showed no tracer uptake in graft center .Both patients with ulnar and humeral reconstruction showed only slightly decreased tracer uptake in bone grafts. 3 phase bone scintigraphy may play a role in the evaluation of bone graft viability by predicting the infection and necrosis. (authors)

  7. Viability and heat resistance of murine norovirus on bread.

    Science.gov (United States)

    Takahashi, Michiko; Takahashi, Hajime; Kuda, Takashi; Kimura, Bon

    2016-01-04

    Contaminated bread was the cause of a large-scale outbreak of norovirus disease in Japan in 2014. Contamination of seafood and uncooked food products by norovirus has been reported several times in the past; however the outbreak resulting from the contamination of bread products was unusual. A few reports on the presence of norovirus on bread products are available; however there have been no studies on the viability and heat resistance of norovirus on breads, which were investigated in this study. ce:italic>/ce:italic> strain 1 (MNV-1), a surrogate for human norovirus, was inoculated directly on 3 types of bread, but the infectivity of MNV-1 on bread samples was almost unchanged after 5days at 20°C. MNV-1 was inoculated on white bread that was subsequently heated in a toaster for a maximum of 2min. The results showed that MNV-1 remained viable if the heating period was insufficient to inactivate. In addition, bread dough contaminated with MNV-1 was baked in the oven. Our results indicated that MNV-1 may remain viable on breads if the heating duration or temperature is insufficient. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Viability of osteocytes in bone autografts harvested for dental implantology

    International Nuclear Information System (INIS)

    Guillaume, Bernard; Gaudin, Christine; Georgeault, Sonia; Mallet, Romain; Basle, Michel F; Chappard, Daniel

    2009-01-01

    Bone autograft remains a very useful and popular way for filling bone defects. In maxillofacial surgery or implantology, it is used to increase the volume of the maxilla or mandible before placing dental implants. Because there is a noticeable delay between harvesting the graft and its insertion in the receiver site, we evaluated the morphologic changes at the light and transmission electron microscopy levels. Five patients having an autograft (bone harvested from the chin) were enrolled in the study. A small fragment of the graft was immediately fixed after harvesting and a second one was similarly processed at the end of the grafting period when bone has been stored at room temperature for a 20 min ± 33 s period in saline. A net increase in the number of osteocyte lacunae filled with cellular debris was observed (+41.5%). However no cytologic alteration could be observed in the remaining osteocytes. The viability of these cells is known to contribute to the success of autograft in association with other less well-identified factors.

  9. Tissue viability monitoring: a multi-sensor wearable platform approach

    Science.gov (United States)

    Mathur, Neha; Davidson, Alan; Buis, Arjan; Glesk, Ivan

    2016-12-01

    Health services worldwide are seeking ways to improve patient care for amputees suffering from diabetes, and at the same time reduce costs. The monitoring of residual limb temperature, interface pressure and gait can be a useful indicator of tissue viability in lower limb amputees especially to predict the occurrence of pressure ulcers. This is further exacerbated by elevated temperatures and humid micro environment within the prosthesis which encourages the growth of bacteria and skin breakdown. Wearable systems for prosthetic users have to be designed such that the sensors are minimally obtrusive and reliable enough to faithfully record movement and physiological signals. A mobile sensor platform has been developed for use with the lower limb prosthetic users. This system uses an Arduino board that includes sensors for temperature, gait, orientation and pressure measurements. The platform transmits sensor data to a central health authority database server infrastructure through the Bluetooth protocol at a suitable sampling rate. The data-sets recorded using these systems are then processed using machine learning algorithms to extract clinically relevant information from the data. Where a sensor threshold is reached a warning signal can be sent wirelessly together with the relevant data to the patient and appropriate medical personnel. This knowledge is also useful in establishing biomarkers related to a possible deterioration in a patient's health or for assessing the impact of clinical interventions.

  10. Agronomic viability of New Zealand spinach and kale intercropping.

    Science.gov (United States)

    Cecílio, Arthur B; Bianco, Matheus S; Tardivo, Caroline F; Pugina, Gabriel C M

    2017-01-01

    The intercropping is a production system that aims to provide increased yield with less environmental impact, due to greater efficiency in the use of natural resources and inputs involved in the production process. An experiment was carried out to evaluate the agronomic viability of kale and New Zealand spinach intercropping as a function of the spinach transplanting time. (0, 14, 28, 42, 56, 70, 84 and 98 days after transplanting of the kale). The total yield (TY) and yield per harvest (YH) of the kale in intercropping did not differ from those obtained in monoculture. The spinach TY was influenced by the transplanting time, the earlier the transplanting, the higher the TY. The spinach YH was not influenced by the transplanting time, but rather by the cultivation system. In intercropping, the spinach YH was 13.5% lower than in monoculture. The intercropping was agronomically feasible, since the land use efficiency index, which was not influenced by the transplanting time, had an average value of 1.71, indicating that the intercropping produced 71% more kale and spinach than the same area in monoculture. Competitiveness coefficient, aggressiveness and yield loss values showed that kale is the dominating species and spinach is the dominated.

  11. Adverse respiratory outcome after premature rupture of membranes before viability.

    Science.gov (United States)

    Verspyck, Eric; Bisson, Violene; Roman, Horace; Marret, Stéphane

    2014-03-01

    To determine whether preterm premature rupture of membranes (PPROM) before 24 weeks is an independent risk factor for poor outcome in preterm neonates. A retrospective comparative cohort study was conducted, including viable premature infants born between 25 and 34-weeks gestation. Each preterm case with early PPROM was matched with two preterm controls of the same gestational age at birth, sex and birth date and who were born spontaneously with intact membranes. Logistic regression was performed to identify independent risk factors associated with composite respiratory and perinatal adverse outcomes for the overall population of preterm infants. Thirty-five PPROM cases were matched with 70 controls. Extreme prematurity (26-28 weeks) was an independent risk factor for composite perinatal adverse outcomes [odds ratio (OR) 43.9; p = 0.001]. Extreme prematurity (OR 42.9; p = 0.001), PPROM (OR 7.1; p = 0.01), male infant (OR 5.2; p = 0.02) and intrauterine growth restriction (IUGR, OR 4.8; p = 0.04) were factors for composite respiratory adverse outcomes. Preterm premature rupture of membranes before viability represents an independent risk factor for composite respiratory adverse outcomes in preterm neonates. Extreme prematurity may represent the main risk factor for both composite respiratory and perinatal adverse outcomes. ©2013 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

  12. Identification and characterization of the Luc2-type luciferase in the Japanese firefly, Luciola parvula, involved in a dim luminescence in immobile stages.

    Science.gov (United States)

    Bessho-Uehara, Manabu; Oba, Yuichi

    2017-09-01

    Nocturnal Japanese fireflies, Luciola parvula, emit from their lanterns a yellow light, one of the most red-shifted colors found among fireflies. Previously, we isolated and characterized two different types of luciferase gene, Luc1 and Luc2, from the fireflies Luciola cruciata and Luciola lateralis; Luc1 is responsible for the green-yellow luminescence of larval and adult lanterns, whereas Luc2 is responsible for the dim greenish glow of eggs and pupal bodies. The biological role of firefly lanterns in adults is related to sexual communication, but why the eggs and pupae glow remains uncertain. In this study, we isolated the gene Luc2 from L. parvula, and compared its expression profiles and enzymatic characteristics with those of Luc1. A semi-quantitative reverse transcription polymerase chain reaction showed that Luc1 was predominantly expressed in larvae, prepupae, pupae and adults, whereas Luc2 was expressed in eggs, prepupae, pupae and adult females. Enzymatic analyses showed that the luminescent color of Luc1 matches the visual sensitivity of L. parvula eyes, whereas that of Luc2 is very different from it. These results suggest that the biological role of Luc2 expressed in immobile stages is not intraspecific communication. Copyright © 2017 John Wiley & Sons, Ltd.

  13. [Establishment of an iRFP and luciferase dual-color fluorescence-traced hepatocellular carcinoma transplantation model in nude mice].

    Science.gov (United States)

    Li, Hongjun; Yang, Tianhua; Huang, Yanping; Liu, Mingzhu; Qin, Zhongqiang; Chu, Fei; Li, Zhenghong; Li, Yonghai

    2017-11-01

    Objective To establish a hepatocellular carcinoma xenograft model in nude mice which could stably express gene and be monitored dynamically. Methods We first constructed the lentiviral particles containing luciferase (Luc) and near-infrared fluorescent protein (iRFP) and puromycin resistance gene, and then transduced them into the HepG2 hepatoma cells. The cell line stably expressing Luc and iRFP genes were screened and inoculated into nude mice to establish xenograft tumor model. Tumor growth was monitored using in vivo imaging system. HE staining and immunohistochemistry were used to evaluate the pathological features and tumorigenic ability. Results HepG2 cells stably expressing iRFP and Luc were obtained; with the engineered cell line, xenograft model was successfully established with the features of proper tumor developing time and high rate of tumor formation as well as typical pathological features as showed by HE staining and immunohistochemistry. Conclusion Hepatocellular carcinoma model in nude mice with the features of stable gene expression and dynamical monitoring has been established successfully with the HepG2-iRFP-Luc cell line.

  14. A dual function fusion protein of Herpes simplex virus type 1 thymidine kinase and firefly luciferase for noninvasive in vivo imaging of gene therapy in malignant glioma.

    Science.gov (United States)

    Söling, Ariane; Theiss, Christian; Jungmichel, Stephanie; Rainov, Nikolai G

    2004-08-04

    BACKGROUND: Suicide gene therapy employing the prodrug activating system Herpes simplex virus type 1 thymidine kinase (HSV-TK)/ ganciclovir (GCV) has proven to be effective in killing experimental brain tumors. In contrast, glioma patients treated with HSV-TK/ GCV did not show significant treatment benefit, most likely due to insufficient transgene delivery to tumor cells. Therefore, this study aimed at developing a strategy for real-time noninvasive in vivo monitoring of the activity of a therapeutic gene in brain tumor cells. METHODS: The HSV-TK gene was fused to the firefly luciferase (Luc) gene and the fusion construct HSV-TK-Luc was expressed in U87MG human malignant glioma cells. Nude mice with subcutaneous gliomas stably expressing HSV-TK-Luc were subjected to GCV treatment and tumor response to therapy was monitored in vivo by serial bioluminescence imaging. Bioluminescent signals over time were compared with tumor volumes determined by caliper. RESULTS: Transient and stable expression of the HSV-TK-Luc fusion protein in U87MG glioma cells demonstrated close correlation of both enzyme activities. Serial optical imaging of tumor bearing mice detected in all cases GCV induced death of tumor cells expressing the fusion protein and proved that bioluminescence can be reliably used for repetitive and noninvasive quantification of HSV-TK/ GCV mediated cell kill in vivo. CONCLUSION: This approach may represent a valuable tool for the in vivo evaluation of gene therapy strategies for treatment of malignant disease.

  15. A simple viability analysis for unicellular cyanobacteria using a new autofluorescence assay, automated microscopy, and ImageJ

    Directory of Open Access Journals (Sweden)

    Schulze Katja

    2011-11-01

    Full Text Available Abstract Background Currently established methods to identify viable and non-viable cells of cyanobacteria are either time-consuming (eg. plating or preparation-intensive (eg. fluorescent staining. In this paper we present a new and fast viability assay for unicellular cyanobacteria, which uses red chlorophyll fluorescence and an unspecific green autofluorescence for the differentiation of viable and non-viable cells without the need of sample preparation. Results The viability assay for unicellular cyanobacteria using red and green autofluorescence was established and validated for the model organism Synechocystis sp. PCC 6803. Both autofluorescence signals could be observed simultaneously allowing a direct classification of viable and non-viable cells. The results were confirmed by plating/colony count, absorption spectra and chlorophyll measurements. The use of an automated fluorescence microscope and a novel ImageJ based image analysis plugin allow a semi-automated analysis. Conclusions The new method simplifies the process of viability analysis and allows a quick and accurate analysis. Furthermore results indicate that a combination of the new assay with absorption spectra or chlorophyll concentration measurements allows the estimation of the vitality of cells.

  16. Evaluation of Escherichia coli viability by flow cytometry: A method for determining bacterial responses to antibiotic exposure.

    Science.gov (United States)

    Boi, Paola; Manti, Anita; Pianetti, Anna; Sabatini, Luigia; Sisti, Davide; Rocchi, Marco Bruno; Bruscolini, Francesca; Galluzzi, Luca; Papa, Stefano

    2015-01-01

    In this study, we check for the presence of specific resistance genes by polymerase chain reaction (PCR) and then we used flow cytometry (FCM) to evaluate antibiotic-induced effects in different strains of Escherichia coli (E. coli). The presence of resistance genes was investigated by PCR in 10 strains of E. coli isolated from Foglia River. Bacterial responses to different antibiotics were also tested with FCM techniques by evaluating both the degree of decrease in viability and the light scatter changes in all of the strains. PCR revealed that only one strain exhibits the presence of one resistance gene. Despite this, analyses of strains using FCM evidenced the presence of viable subpopulations after antibiotic treatment. Furthermore, analyses of scatter signals revealed profound changes in the Forward Scatter and Side Scatter of the bacterial populations as a consequence of antibiotic exposure, confirming the viability and membrane potential data. The riverine strains were in general less sensitive to antibiotics than the reference strain (ATCC 25922). Antibiotic resistance is a widespread phenomena. The multiparametric approach based on FCM used in this study, providing results about different aspects (cell viability, membrane potential, light scatter changes), may overcome the limitation of PCR and could represent an adequate method for the evaluation of bacteria responses to antibiotic exposure. © 2014 International Clinical Cytometry Society.

  17. Moringa oleifera's Nutritious Aqueous Leaf Extract Has Anticancerous Effects by Compromising Mitochondrial Viability in an ROS-Dependent Manner.

    Science.gov (United States)

    Madi, Niveen; Dany, Mohammed; Abdoun, Salah; Usta, Julnar

    2016-01-01

    Moringa oleifera (MO) is an important dietary component for many populations in West Africa and the Indian subcontinent. In addition to its highly nutritious value, almost all parts of this plant have been widely used in folk medicine in curing infectious, cardiovascular, gastrointestinal, hepatic, and other diseases. Evidence-based research supported its versatile medicinal properties; however, more rigorous research is required to establish it in cancer therapy. As such, in this study we aim to investigate the in vitro anticancerous effect of Moringa oleifera's aqueous leaf extract. Moringa extract was prepared by soaking pulverized leaves in hot water mimicking the people's mode of the leaf drink preparation. Several assays were used to study the effect of different percentage concentrations of the extract on viability of A549 cells; levels of adenosine triphosphate (ATP), reactive oxygen species (ROS), and glutathione (GSH) generated; as well as percentage of lactate dehydrogenase (LDH) released at different time points. In addition to mitochondrial membrane potential, apoptotic events were assessed using western blotting for apoptotic markers and immunoflourescent flourescent labeled inhibitor of caspases (FLICA) assay. MO extract treatment resulted in a significant decrease in mitochondrial membrane potential (1 hour) and ATP levels (3 hours), followed by an increase in (6 hours) ROS, caspase activation, proapoptotic proteins expression (p53, SMAC/Diablo, AIF), and PARP-1 cleavage. This eventually resulted in decreased GSH levels and a decrease in viability. The cytotoxic effect was prevented upon pretreatment with antioxidant N-acetyl-cysteine. MO decreased as well the viability of HepG2, CaCo2, Jurkat, and HEK293 cells. Our findings identify a plant extract with an anticancerous effect on cancer cell lines. MO extract exerts its cytotoxic effect in A549 cancer cells by affecting mitochondrial viability and inducing apoptosis in an ROS-dependent manner.

  18. Identifying rapidly parasiticidal anti-malarial drugs using a simple and reliable in vitro parasite viability fast assay.

    Science.gov (United States)

    Linares, María; Viera, Sara; Crespo, Benigno; Franco, Virginia; Gómez-Lorenzo, María G; Jiménez-Díaz, María Belén; Angulo-Barturen, Íñigo; Sanz, Laura María; Gamo, Francisco-Javier

    2015-11-05

    The emergence of Plasmodium falciparum resistance to artemisinins threatens to undermine the effectiveness of artemisinin-based combination anti-malarial therapy. Developing suitable drugs to replace artemisinins requires the identification of new compounds that display rapid parasite killing kinetics. However, no current methods fully meet the requirements to screen large compound libraries for candidates with such properties. This study describes the development and validation of an in vitro parasite viability fast assay for identifying rapidly parasiticidal anti-malarial drugs. Parasite killing kinetics were determined by first culturing unlabelled erythrocytes with P. falciparum in the presence of anti-malarial drugs for 24 or 48 h. After removing the drug, samples were added to erythrocytes pre-labelled with intracellular dye to allow their subsequent identification. The ability of viable parasites to re-establish infection in labelled erythrocytes could then be detected by two-colour flow cytometry after tagging of parasite DNA. Thus, double-stained erythrocytes (with the pre-labelled intracellular dye and the parasite DNA dye) result only after establishment of new infections by surviving parasites. The capacity of the test anti-malarial drugs to eliminate viable parasites within 24 or 48 h could, therefore, be determined. The parasite viability fast assay could be completed within 48 h following drug treatment and distinguished between rapidly parasiticidal anti-malarial drugs versus those acting more slowly. The assay was validated against ten standard anti-malarial agents with known properties and results correlated well with established methods. An abbreviated assay, suitable for adaption to medium-high throughput screening, was validated and applied against a set of 20 compounds retrieved from the publically available Medicines for Malaria Venture 'Malaria Box'. The quantification of new infections to determine parasite viability offers important

  19. Viability of chondrocytes seeded onto a collagen I/III membrane for matrix-induced autologous chondrocyte implantation.

    Science.gov (United States)

    Hindle, Paul; Hall, Andrew C; Biant, Leela C

    2014-11-01

    Cell viability is crucial for effective cell-based cartilage repair. The aim of this study was to determine the effect of handling the membrane during matrix-induced autologous chondrocyte implantation surgery on the viability of implanted chondrocytes. Images were acquired under five conditions: (i) Pre-operative; (ii) Handled during surgery; (iii) Cut edge; (iv) Thumb pressure applied; (v) Heavily grasped with forceps. Live and dead cell stains were used. Images were obtained for cell counting and morphology. Mean cell density was 6.60 × 10(5) cells/cm(2) (5.74-7.11 × 10(5) ) in specimens that did not have significant trauma decreasing significantly in specimens that had been grasped with forceps (p < 0.001) or cut (p = 0.004). Cell viability on delivery grade membrane was 75.1%(72.4-77.8%). This dropped to 67.4%(64.1-69.7%) after handling (p = 0.002), 56.3%(51.5-61.6%) after being thumbed (p < 0.001) and 28.8%(24.7-31.2%) after crushing with forceps (p < 0.001). When cut with scissors there was a band of cell death approximately 275 µm in width where cell viability decreased to 13.7%(10.2-18.2%, p < 0.001). Higher magnification revealed cells without the typical rounded appearance of chondrocytes. We found that confocal laser-scanning microscope (CLSM) can be used to quantify and image the fine morphology of cells on a matrix-induced autologous chondrocyte implantation (MACI) membrane. Careful handling of the membrane is essential to minimise chondrocyte death during surgery. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  20. Ginseng (Panax quinquefolius and Licorice (Glycyrrhiza uralensis Root Extract Combinations Increase Hepatocarcinoma Cell (Hep-G2 Viability

    Directory of Open Access Journals (Sweden)

    David G. Popovich

    2011-01-01

    Full Text Available The combined cytoactive effects of American ginseng (Panax quinquefolius and licorice (Glycyrrhiza uralensis root extracts were investigated in a hepatocarcinoma cell line (Hep-G2. An isobolographic analysis was utilized to express the possibility of synergistic, additive or antagonistic interaction between the two extracts. Both ginseng and licorice roots are widely utilized in traditional Chinese medicine preparations to treat a variety of ailments. However, the effect of the herbs in combination is currently unknown in cultured Hep-G2 cells. Ginseng (GE and licorice (LE extracts were both able to reduce cell viability. The LC50 values, after 72 h, were found to be 0.64 ± 0.02 mg/mL (GE and 0.53 ± 0.02 mg/mL (LE. An isobologram was plotted, which included five theoretical LC50s calculated, based on the fixed fraction method of combination ginseng to licorice extracts to establish a line of additivity. All combinations of GE to LE (1/5, 1/3, 1/2, 2/3, 4/5 produced an effect on Hep-G2 cell viability but they were all found to be antagonistic. The LC50 of fractions 1/3, 1/2, 2/3 were 23%, 21% and 18% above the theoretical LC50. Lactate dehydrogenase release indicated that as the proportion of GE to LE increased beyond 50%, the influence on membrane permeability increased. Cell-cycle analysis showed a slight but significant arrest at the G1 phase of cell cycle for LE. Both GE and LE reduced Hep-G2 viability independently; however, the combinations of both extracts were found to have an antagonistic effect on cell viability and increased cultured Hep-G2 survival.

  1. 75 FR 42474 - The Future of Aviation Advisory Committee (FAAC) Subcommittee on Competitiveness and Viability...

    Science.gov (United States)

    2010-07-21

    ... of Aviation Advisory Committee (FAAC) Subcommittee on Competitiveness and Viability; Notice of... Transportation, announces the second meeting of the FAAC Subcommittee on Competitiveness and Viability, which... recommendations to the Secretary of Transportation to ensure the competitiveness of the U.S. aviation industry and...

  2. Effect of chlorinated ethene conversion on viability and activity of Methylosinus trichosporium OB3b

    NARCIS (Netherlands)

    van Hylckama Vlieg, Johan E.T.; Koning, Wim de; Janssen, Dick B.

    1997-01-01

    The effect of transformation of chlorinated ethenes on the cell viability of Methylosinus trichosporium OB3b was investigated. A comparison of the loss of viability with the decrease in transformation rates shelved that for the monooxygenase-mediated transformation of all chlorinated ethenes except

  3. Stochastic modelling of the economic viability of on-farm co-digestion of pig manure and food waste in Ireland

    International Nuclear Information System (INIS)

    Dennehy, C.; Lawlor, P.G.; Gardiner, G.E.; Jiang, Y.; Shalloo, L.; Zhan, X.

    2017-01-01

    Highlights: •Assessed economic viability of on-farm manure mono- and co-digestion. •Assessed three farm sizes: 521 sows; 2607 sows; and 5214 sows. •Mono-digestion of manure alone not economically viable. •Co-digestion viable on small farms as food waste likely to be sourced. •Viability on larger farms dependent on securing sufficient food waste. -- Abstract: The majority of studies analysing the economic potential of biogas systems utilise deterministic models to assess the viability of a system using fixed inputs. However, changes in market conditions can significantly affect the viability of biogas plants, and need to be accounted for. This study assessed the economic potential of undertaking on-farm anaerobic co-digestion of food waste (FW) and pig manure (PM) using both deterministic and stochastic modelling approaches. The financial viability of three co-digestion plants sized to treat PM generated from 521, 2607 and 5214 sow integrated units was assessed. Under current market conditions the largest co-digestion scenario modelled was found to be unviable. Stochastic modelling of four key input variables (FW availability, renewable electricity tariff, gate fees and digestate disposal costs) was undertaken to assess the sensitivity of project viability to changes in market conditions. Due to the high likelihood of accessing sufficient FW, the smallest co-digestion scenario was found to be the least sensitive to any future changes in market conditions. Due to its potential to treat greater amounts of FW than the smallest scenario, a co-digestion plant designed for a 2607 sow farm had the highest revenue generating potential under optimal market conditions; however, it was more sensitive to changes in FW availability than the smaller scenario. This study illustrates the need for farm-based biogas plant projects to secure long-term, stable supplies of co-substrates and to size plants’ capacity based on the availability of the co-substrates which drive

  4. Rapid, portable and cost-effective yeast cell viability and concentration analysis using lensfree on-chip microscopy and machine learning

    KAUST Repository

    Feizi, Alborz; Zhang, Yibo; Greenbaum, Alon; Guziak, Alex; Luong, Michelle; Chan, Raymond Yan Lok; Berg, Brandon; Ozkan, Haydar; Luo, Wei; Wu, Michael; Wu, Yichen; Ozcan, Aydogan

    2016-01-01

    and cost-effective automatic yeast analysis platform (AYAP), which can rapidly measure cell concentration and viability. AYAP is based on digital in-line holography and on-chip microscopy and rapidly images a large field-of-view of 22.5 mm2. This lens

  5. Optimising the economic viability of grid-connected photovoltaic systems

    International Nuclear Information System (INIS)

    Mondol, Jayanta Deb; Yohanis, Yigzaw G; Norton, Brian

    2009-01-01

    The impact of photovoltaic (PV) array size, orientation, inclination, load profile, electricity buying price, feed-in tariffs, PV/inverter sizing ratio ('sizing ratio') and PV/inverter cost ratio ('cost ratio') on the economic viability of a grid-connected PV system was investigated using a validated TRNSYS simulation model. The results showed that the fractional load met directly by a PV system depends on matching between PV supply and building load profile, sizing ratio and PV inclination. The profitability of a grid-connected PV system increases if the PV system is sized to reduce excess PV electrical energy fed to the grid when the feed-in tariff is lower than electricity buying price. The effect of feed-in tariffs on PV saving for selected European countries has been shown. The cost of the PV electricity depends on sizing ratio, PV and inverter lifetimes, cost ratio, PV inclination and financial parameters. The effect of cost ratio on the optimum PV/inverter sizing ratio is less significant when the cost ratio lies within 7-11. The minimum PV electricity cost at low and high insolation conditions were obtained for sizing ratios of 1.6 and 1.2, respectively. The lowest PV electricity cost was found for surface slopes within 30-40 for the selected European locations. The PV electricity cost for cost ratio of 5 and 13 varied from 0.44-0.85 EURkWh -1 to 0.38-0.76 EURkWh -1 , respectively within high to low insolation conditions when the PV module unit cost, market discount rate, PV size, PV lifetime and inverter lifetime were assumed to be 6.5 EURW p -1 , 3%, 13 kW p , 20 years and 10 years, respectively. (author)

  6. Myocardial viability assessed by Tl-201 SPECT. Redistribution versus reinjection

    International Nuclear Information System (INIS)

    Chalela, William Azem; Pimentel, Flavio Ferrarini de Oliveira; Uchida, Augusto Hiroshi; Bottega, Augusto; Ramires, Jose Antonio Franchine; Izaki, Marisa; Moraes, Aguinaldo Pereira; Soares Junior, Jose; Giorgi, Maria C. Pinto; Moffa, Paulo Jorge; Bellotti, Giovanni; Giovanni Guido Cerri; Meneghetti, Jose Claudio

    1994-01-01

    The purpose of this study was to verify if a third series of images acquired by reinjection thallium-201, 24 h after conventional myocardial perfusion with the radioisotope, improves the identification of myocardial viability segments. The methods: we studied 30 patients, mean age 57.7 ±9.4 years, with old myocardial infarction using thallium (Tl)-201 SPECT, and we obtained three series of images (stress, redistribution after 4 h and reinjection after 24 h. Cardiac images were divided in 5 segments (apical, lateral, anterior, septal and inferior) and each one received a value by a score system according to the Tl-201 myocardial uptake (0=normal uptake; 1=mild hypoperfusion; 2=moderate hypoperfusion; 3=severe hypoperfusion or no myocardial uptake). We considered viable myocardium when the uptake of Tl-201 in the segment related to te myocardial infarction increases at least 1 point in two different axis of Tl-201 SPECT. The results: seven (23,3%) patients demonstrated increase of Tl-201 uptake only at reinjection images, showing a high efficacy of the method. Nine (30%) patients showed persistent hypoperfusion at all series of images suggesting only fibrosis in the are related to the infarction. Fourteen (46,7%) patients showed increase of Tl-201 concentration at redistribution images; among these patients, six showed improvement of myocardial uptake at reinjection. This condition was interpreted as regional chronic ischemic process: hibernating myocardium. The conclusion was that Tl-201 hypoperfusion at redistribution images without significant changes in relation to the stress images do not represent fibrosis at all. The reinjection technic was better than conventional redistribution in the detection of viable myocardium. This data allows a better therapeutic orientation. (author)

  7. Aspergillus fumigatus viability drives allergic responses to inhaled conidia.

    Science.gov (United States)

    Nayak, Ajay P; Croston, Tara L; Lemons, Angela R; Goldsmith, W T; Marshall, Nikki B; Kashon, Michael L; Germolec, Dori R; Beezhold, Donald H; Green, Brett J

    2018-04-13

    Aspergillus fumigatus induced allergic airway disease has been shown to involve conidial germination in vivo but the immunological mechanisms remain uncharacterized. A subchronic murine exposure model was used to examine the immunological mediators that are regulated in response to either culturable or non-culturable A. fumigatus conidia. Female B6C3F1/N mice were repeatedly dosed via inhalation with 1 x 105 viable or heat inactivated conidia (HIC), twice a week for 13 weeks (26 exposures). Control mice inhaled HEPA-filtered air. The influence of A. fumigatus conidial germination on the pulmonary immunopathological outcomes was evaluated by flow cytometry analysis of cellular infiltration in the airways, assessment of lung mRNA expression, and quantitative proteomics and histopathology of whole lung tissue. Repeated inhalation of viable conidia, but not HIC, resulted in allergic inflammation marked by vascular remodeling, extensive eosinophilia, and accumulation of alternatively activated macrophages (AAMs) in the murine airways. More specifically, mice that inhaled viable conidia resulted in a mixed TH1 and TH2 (IL-13) cytokine response. Recruitment of eosinophils corresponded with increased Ccl11 transcripts. Furthermore, genes associated with M2 or alternatively activated macrophage polarization (e.g. Arg1, Chil3 and Retnla) were significantly upregulated in viable A. fumigatus exposed mice. In mice inhaling HIC, CD4+ T cells expressing IFN-γ (TH1) dominated the lymphocytic infiltration. Quantitative proteomics of the lung revealed metabolic reprogramming accompanied by mitochondrial dysfunction and endoplasmic reticulum stress stimulated by oxidative stress from repetitive microbial insult. Our studies demonstrate that A. fumigatus conidial viability in vivo is critical to the immunopathological presentation of chronic fungal allergic disease. Copyright © 2018. Published by Elsevier Inc.

  8. COST IMPACT OF ROD CONSOLIDATION ON THE VIABILITY ASSESSMENT DESIGN

    International Nuclear Information System (INIS)

    D. Lancaster

    1999-01-01

    The cost impact to the Civilian Radioactive Waste Management System of using rod consolidation is evaluated. Previous work has demonstrated that the fuel rods of two assemblies can be packed into a canister that can fit into the same size space as that used to store a single assembly. The remaining fuel assembly hardware can be compacted into the same size canisters with a ratio of 1 hardware canister per each 6 to 12 assemblies. Transportation casks of the same size as currently available can load twice the number of assemblies by placing the compacted assemblies in the slots currently designed for a single assembly. Waste packages similarly could contain twice the number of assemblies; however, thermal constraints would require considering either a low burnup or cooling. The analysis evaluates the impact of rod consolidation on CRWMS costs for consolidation at prior to transportation and for consolidation at the Monitored Geological Repository surface facility. For this study, no design changes were made to either the transport casks or waste packages. Waste package designs used for the Viability Assessment design were employed but derated to make the thermal limits. A logistics analysis of the waste was performed to determine the number of each waste package with each loading. A review of past rod consolidation experience found cost estimates which range from $10/kgU to $32/kgU. $30/kgU was assumed for rod consolidation costs prior to transportation. Transportation cost savings are about $17/kgU and waste package cost savings are about $21/kgU. The net saving to the system is approximately $500 million if the consolidation is performed prior to transportation. If consolidation were performed at the repository surface facilities, it would cost approximately $15/kgU. No transportation savings would be realized. The net savings for consolidation at the repository site would be about $400 million dollars

  9. A model to determine the economic viability of water fluoridation.

    Science.gov (United States)

    Kroon, Jeroen; van Wyk, Philippus Johannes

    2012-01-01

    In view of concerns expressed by South African local authorities the aim of this study was to develop a model to determine whether water fluoridation is economically viable to reduce dental caries in South Africa. Microsoft Excel software was used to develop a model to determine economic viability of water fluoridation for 17 water providers from all nine South African provinces. Input variables for this model relate to chemical cost, labor cost, maintenance cost of infrastructure, opportunity cost, and capital depreciation. The following output variables were calculated to evaluate the cost of water fluoridation: per capita cost per year, cost-effectiveness and cost-benefit. In this model it is assumed that the introduction of community water fluoridation can reduce caries prevalence by an additional 15 percent and that the savings in cost of treatment will be equal to the average fee for a two surface restoration. Water providers included in the study serve 53.5 percent of the total population of South Africa. For all providers combined chemical cost contributes 64.5 percent to the total cost, per capita cost per year was $0.36, cost-effectiveness was calculated as $11.41 and cost-benefit of the implementation of water fluoridation was 0.34. This model confirmed that water fluoridation is an economically viable option to prevent dental caries in South African communities, as well as conclusions over the last 10 years that water fluoridation leads to significant cost savings and remains a cost-effective measure for reducing dental caries, even when the caries-preventive effectiveness is modest. © 2012 American Association of Public Health Dentistry.

  10. Detection and viability of Lactococcus lactis throughout cheese ripening.

    Directory of Open Access Journals (Sweden)

    Marianna Ruggirello

    Full Text Available Recent evidences highlighted the presence of Lactococcus lactis during late cheese ripening. For this reason, the role of this microorganism, well known as dairy starter, should be reconsidered throughout cheese manufacturing and ripening. Thus, the main objective of this study was to develop a RT-qPCR protocol for the detection, quantification and determination of the viability of L. lactis in ripened cheese samples by direct analysis of microbial nucleic acids. Standard curves were constructed for the specific quantification of L. lactis in cheese matrices and good results in terms of selectivity, correlation coefficient and efficiency were obtained. Thirty-three ripened cheeses were analyzed and, on the basis of RNA analysis, twelve samples showed 106 to 108 CFU of L. lactis per gram of product, thirteen from 103 to 105 CFU/g, and in eight cheeses, L. lactis was not detected. Traditional plating on M17 medium led to loads ranging from 105 to 109 CFU/g, including the cheese samples where no L. lactis was found by RT-qPCR. From these cheeses, none of the colonies isolated on M17 medium was identified as L. lactis species. These data could be interpreted as a lack of selectivity of M17 medium where colony growth is not always related to lactococcal species. At the same time, the absence or low abundance of L. lactis isolates on M17 medium from cheese where L. lactis was detected by RT-qPCR support the hypothesis that L. lactis starter populations are mainly present in viable but not culturable state during ripening and, for this reason, culture-dependent methods have to be supplemented with direct analysis of cheese.

  11. Detection and Viability of Lactococcus lactis throughout Cheese Ripening

    Science.gov (United States)

    Cocolin, Luca

    2014-01-01

    Recent evidences highlighted the presence of Lactococcus lactis during late cheese ripening. For this reason, the role of this microorganism, well known as dairy starter, should be reconsidered throughout cheese manufacturing and ripening. Thus, the main objective of this study was to develop a RT-qPCR protocol for the detection, quantification and determination of the viability of L. lactis in ripened cheese samples by direct analysis of microbial nucleic acids. Standard curves were constructed for the specific quantification of L. lactis in cheese matrices and good results in terms of selectivity, correlation coefficient and efficiency were obtained. Thirty-three ripened cheeses were analyzed and, on the basis of RNA analysis, twelve samples showed 106 to 108 CFU of L. lactis per gram of product, thirteen from 103 to 105 CFU/g, and in eight cheeses, L. lactis was not detected. Traditional plating on M17 medium led to loads ranging from 105 to 109 CFU/g, including the cheese samples where no L. lactis was found by RT-qPCR. From these cheeses, none of the colonies isolated on M17 medium was identified as L. lactis species. These data could be interpreted as a lack of selectivity of M17 medium where colony growth is not always related to lactococcal species. At the same time, the absence or low abundance of L. lactis isolates on M17 medium from cheese where L. lactis was detected by RT-qPCR support the hypothesis that L. lactis starter populations are mainly present in viable but not culturable state during ripening and, for this reason, culture-dependent methods have to be supplemented with direct analysis of cheese. PMID:25503474

  12. Rapid-viability PCR method for detection of live, virulent Bacillus anthracis in environmental samples.

    Science.gov (United States)

    Létant, Sonia E; Murphy, Gloria A; Alfaro, Teneile M; Avila, Julie R; Kane, Staci R; Raber, Ellen; Bunt, Thomas M; Shah, Sanjiv R

    2011-09-01

    In the event of a biothreat agent release, hundreds of samples would need to be rapidly processed to characterize the extent of contamination and determine the efficacy of remediation activities. Current biological agent identification and viability determination methods are both labor- and time-intensive such that turnaround time for confirmed results is typically several days. In order to alleviate this issue, automated, high-throughput sample processing methods were developed in which real-time PCR analysis is conducted on samples before and after incubation. The method, referred to as rapid-viability (RV)-PCR, uses the change in cycle threshold after incubation to detect the presence of live organisms. In this article, we report a novel RV-PCR method for detection of live, virulent Bacillus anthracis, in which the incubation time was reduced from 14 h to 9 h, bringing the total turnaround time for results below 15 h. The method incorporates a magnetic bead-based DNA extraction and purification step prior to PCR analysis, as well as specific real-time PCR assays for the B. anthracis chromosome and pXO1 and pXO2 plasmids. A single laboratory verification of the optimized method applied to the detection of virulent B. anthracis in environmental samples was conducted and showed a detection level of 10 to 99 CFU/sample with both manual and automated RV-PCR methods in the presence of various challenges. Experiments exploring the relationship between the incubation time and the limit of detection suggest that the method could be further shortened by an additional 2 to 3 h for relatively clean samples.

  13. Influence of low temperature and frost duration on Phytophthora alni subsp. alni viability

    Energy Technology Data Exchange (ETDEWEB)

    Cerny, K.; Filipova, N.; Strnadova, V.

    2012-11-01

    Limits on the survival of P. alni subsp. alni (PAA) due to low temperature can be expected based on previously published laboratory and field studies. This study presents a laboratory experiment to test the influence of low temperature and frost duration on PAA viability. Ten PAA isolates were incubated at different temperatures (-0.1, -2.5, -5.0, -7.5, and -10.0 degree centigrade) and frost durations (0 - 7, 14, 21, and 28 days). A regression analysis confirmed the significant influence of both factors (low temperature and frost duration, and their interaction) on the survival of the pathogen under laboratory conditions. The survival and failure time analysis showed that the survival of the pathogen differs significantly after mild frost (all the isolates tested survived temperatures between -0.1 and -5.0 degree centigrade during the entire testing period) and heavy frost (the pathogen died after 21 days of incubation at -7.5 degree centigrade and after 2 days at -10.0 degree centigrade). Moreover, the viability of the pathogen decreased significantly if the temperature of -5.0 degree centigrade was maintained for at least 1 week and the temperature of -7.5 degree centigrade persisted in laboratory conditions for at least 4 days. The results of the study proved the pathogen to be very sensitive to heavy frost. The low-temperature limits for PAA occur regularly in Central Europe in January. It is probable that these temperatures can reduce PAA populations in diseased black alder stems. The climate change characterised by increases in the lowest minimum winter temperatures in Central Europe (as hypothesised by IPCC) may pose a significant risk for affected alder population in the area. (Author) 21 refs.

  14. Emerging economic viability of grid defection in a northern climate using solar hybrid systems

    International Nuclear Information System (INIS)

    Kantamneni, Abhilash; Winkler, Richelle; Gauchia, Lucia; Pearce, Joshua M.

    2016-01-01

    High demand for photovoltaic (PV), battery, and small-scale combined heat and power (CHP) technologies are driving a virtuous cycle of technological improvements and cost reductions in off-grid electric systems that increasingly compete with the grid market. Using a case study in the Upper Peninsula of Michigan, this paper quantifies the economic viability of off-grid PV+battery+CHP adoption and evaluates potential implications for grid-based utility models. The analysis shows that already some households could save money by switching to a solar hybrid off-grid system in comparison to the effective electric rates they are currently paying. Across the region by 2020, 92% of seasonal households and ~75% of year-round households are projected to meet electricity demands with lower costs. Furthermore, ~65% of all Upper Peninsula single-family owner-occupied households will both meet grid parity and be able to afford the systems by 2020. The results imply that economic circumstances could spur a positive feedback loop whereby grid electricity prices continue to rise and increasing numbers of customers choose alternatives (sometimes referred to as a “utility death spiral”), particularly in areas with relatively high electric utility rates. Utility companies and policy makers must take the potential for grid defection seriously when evaluating energy supply strategies. - Highlights: •Quantifies the economic viability of off-grid hybrid photovoltaic (PV) systems. •PV is backed up with batteries and combined heat and power (CHP). •Case study in Michigan by household size (energy demand) and income. •By 2020, majority of single-family owner-occupied households can defect. •To prevent mass-scale grid defection policies needed for grid-tied PV systems.

  15. Diversity in the dry land mixed system and viability of dairy sheep farming

    Directory of Open Access Journals (Sweden)

    Jose Rivas

    2015-05-01

    Full Text Available Castilla La Mancha is a Spanish region where sheep farming system is traditionally pasture-based. Recently, this territory has undergone a recession of dairy sheep activity, which changed the type and intensity of land utilization and led to environmental and landscape degradation. The present study analyzed the diversity and viability of dairy sheep of mixed systems. Multivariate analysis was conducted on 157 dairy sheep farms, factor analysis selected 3 productivity factors (level of intensification, land use, size and family labour, and cluster analysis classified farms into three groups. Group 1, smallholders – with the smallest size (405.5 ewes and 564.7 ha, lowest area in ownership (1.5%, and agriculture activity (6.5% crops area: family farms (90.8% highly dependent on external inputs. Group 2, large-scale farms (1058.7 ewes and 1755.1 ha – with the lowest stocking rate (0.14 livestock unit/ha and productivity: nonfamily farms (39.1% with low area in ownership (4.1% and agriculture activity (7.6%. Group 3, mixed-technified – with the highest levels of technology and least use of family labour (27.0%: large-scale farms (1387.4 ewes and 955.8 ha, combining milk production with agricultural activities (55.7% crops area, with the highest area in ownership (63.1% and the best productivity performance. In conclusion, the dry land mixed system of Castilla La Mancha showed diversity of farms. Improving viability requires a systemic approach where the key tool is grazing, allowing the mixed system to be consolidated as a model that enhances the positive impact of livestock on the environment in the Mediterranean basin.

  16. Consequences of team charter quality: Teamwork mental model similarity and team viability in engineering design student teams

    Science.gov (United States)

    Conway Hughston, Veronica

    Since 1996 ABET has mandated that undergraduate engineering degree granting institutions focus on learning outcomes such as professional skills (i.e. solving unstructured problems and working in teams). As a result, engineering curricula were restructured to include team based learning---including team charters. Team charters were diffused into engineering education as one of many instructional activities to meet the ABET accreditation mandates. However, the implementation and execution of team charters into engineering team based classes has been inconsistent and accepted without empirical evidence of the consequences. The purpose of the current study was to investigate team effectiveness, operationalized as team viability, as an outcome of team charter implementation in an undergraduate engineering team based design course. Two research questions were the focus of the study: a) What is the relationship between team charter quality and viability in engineering student teams, and b) What is the relationship among team charter quality, teamwork mental model similarity, and viability in engineering student teams? Thirty-eight intact teams, 23 treatment and 15 comparison, participated in the investigation. Treatment teams attended a team charter lecture, and completed a team charter homework assignment. Each team charter was assessed and assigned a quality score. Comparison teams did not join the lecture, and were not asked to create a team charter. All teams completed each data collection phase: a) similarity rating pretest; b) similarity posttest; and c) team viability survey. Findings indicate that team viability was higher in teams that attended the lecture and completed the charter assignment. Teams with higher quality team charter scores reported higher levels of team viability than teams with lower quality charter scores. Lastly, no evidence was found to support teamwork mental model similarity as a partial mediator of the team charter quality on team viability

  17. The effect of CO2 laser beam welded AISI 316L austenitic stainless steel on the viability of fibroblast cells, in vitro.

    Science.gov (United States)

    Köse, Ceyhun; Kaçar, Ramazan; Zorba, Aslı Pınar; Bağırova, Melahat; Allahverdiyev, Adil M

    2016-03-01

    It has been determined by the literature research that there is no clinical study on the in vivo and in vitro interaction of the cells with the laser beam welded joints of AISI 316L biomaterial. It is used as a prosthesis and implant material and that has adequate mechanical properties and corrosion resistance characteristics. Therefore, the interaction of the CO2 laser beam welded samples and samples of the base metal of AISI 316L austenitic stainless steel with L929 fibroblast cells as an element of connective tissue under in vitro conditions has been studied. To study the effect of the base metal and the laser welded test specimens on the viability of the fibroblast cells that act as an element of connective tissues in the body, they were kept in DMEMF-12 medium for 7, 14, 28 days and 18 months. The viability study was experimentally studied using the MTT method for 7, 14, 28 days. In addition, the direct interaction of the fibroblast cells seeded on 6 different plates with the samples was examined with an inverted microscope. The MTT cell viability experiment was repeated on the cells that were in contact with the samples. The statistical relationship was analyzed using a Tukey test for the variance with the GraphPad statistics software. The data regarding metallic ion release were identified with the ICP-MS method after the laser welded and main material samples were kept in cell culture medium for 18 months. The cell viability of the laser welded sample has been detected to be higher than that of the base metal and the control based on 7th day data. However, the laser welded sample's viability of the fibroblast cells has diminished by time during the test period of 14 and 28 days and base metal shows better viability when compared to the laser welded samples. On the other hand, the base metal and the laser welded sample show better cell viability effect when compared to the control group. According to the ICP-MS results of the main material and laser welded

  18. Impact of age and diagnosis on viability during centrifugation and cryopreservation of peripheral blood stem cell products.

    Science.gov (United States)

    Civriz Bozdag, S; Bay, M; Ayyıldız, E; Topcuoglu, P; Ilhan, O

    2012-08-01

    The viability of the hematopoietic stem cells infused to the patient is important for transplant outcome. We evaluated 31 peripheral blood stem cell product collected from 15 patients. We aimed to check the viabilities of the cells from patients with different age and diagnosis, in different stages of the cryopreservation procedure. We showed a markedly decreased viability rate after centrifugation and addition of DMSO. Percentages of viabilities were similar between young and old patients in each step. Type of hematological malignancy did not make a significant influence on the viability. High speed centrifugation has a negative impact on the viability. Copyright © 2012 Elsevier Ltd. All rights reserved.

  19. Population viability analysis: using a modeling tool to assess the viability of tapir populations in fragmented landscapes.

    Science.gov (United States)

    Medici, Emília Patrícia; Desbiez, Arnaud Leonard Jean

    2012-12-01

    A population viability analysis (PVA) was conducted of the lowland tapir populations in the Atlantic Forest of the Pontal do Paranapanema region, Brazil, including Morro do Diabo State Park (MDSP) and surrounding forest fragments. Results from the model projected that the population of 126 tapirs in MDSP is likely to persist over the next 100 years; however, 200 tapirs would be required to maintain a viable population. Sensitivity analysis showed that sub-adult mortality and adult mortality have the strongest influence on the dynamics of lowland tapir populations. High road-kill has a major impact on the MDSP tapir population and can lead to population extinction. Metapopulation modeling showed that dispersal of tapirs from MDSP to the surrounding fragments can be detrimental to the overall metapopulation, as fragments act as sinks. Nevertheless, the model showed that under certain conditions the maintenance of the metapopulation dynamics might be determinant for the persistence of tapirs in the region, particularly in the smaller fragments. The establishment of corridors connecting MDSP to the forest fragments models resulted in an increase in the stochastic growth rate, making tapirs more resilient to threats and catastrophes, but only if rates of mortality were not increased when using corridors. The PVA showed that the conservation of tapirs in the Pontal region depends on: the effective protection of MDSP; maintenance and, whenever possible, enhancement of the functional connectivity of the landscape, reducing mortality during dispersal and threats in the unprotected forest fragments; and neutralization of all threats affecting tapirs in the smaller forest fragments. © 2012 Wiley Publishing Asia Pty Ltd, ISZS and IOZ/CAS.

  20. Framework for Assessing Viability of Threatened and Endangered Chinook Salmon and Steelhead in the Sacramento–San Joaquin Basin

    Directory of Open Access Journals (Sweden)

    Steven T. Lindley

    2007-02-01

    Full Text Available Protected evolutionarily significant units (ESUs of salmonids require objective and measurable criteria for guiding their recovery. In this report, we develop a method for assessing population viability and two ways to integrate these population-level assessments into an assessment of ESU viability. Population viability is assessed with quantitative extinction models or criteria relating to population size, population growth rate, the occurrence of catastrophic declines, and the degree of hatchery influence. ESU viability is assessed by examining the number and distribution of viable populations across the landscape and their proximity to sources of catastrophic disturbance. Central Valley spring-run and winter-run Chinook salmon ESUs are not currently viable, according to the criteria-based assessment. In both ESUs, extant populations may be at low risk of extinction, but these populations represent a small portion of the historical ESUs, and are vulnerable to catastrophic disturbance. The winter-run Chinook salmon ESU, in the extreme case, is represented by a single population that spawns outside of its historical spawning range. We are unable to assess the status of the Central Valley steelhead ESU with our framework because almost all of its roughly 80 populations are classified as data deficient. The few exceptions are those populations with a closely associated hatchery, and the naturally-spawning fish in these streams are at high risk of extinction. Population monitoring in this ESU is urgently needed. Global and regional climate change poses an additional risk to the survival of salmonids in the Central Valley. A literature review suggests that by 2100, mean summer temperatures in the Central Valley region may increase by 2-8°C, precipitation will likely shift to more rain and less snow, with significant declines in total precipitation possible, and hydrographs will likely change, especially the the southern Sierra Nevada mountains

  1. Space-time analysis of gravitropism in etiolated Arabidopsis hypocotyls using bioluminescence imaging of the IAA19 promoter fusion with a destabilized luciferase reporter.

    Science.gov (United States)

    Yamamoto, Kotaro T; Watahiki, Masaaki K; Matsuzaki, Jun; Satoh, Soichirou; Shimizu, Hisayo

    2017-07-01

    Imaging analysis was carried out during the gravitropic response of etiolated Arabidopsis hypocotyls, using an IAA19 promoter fusion of destabilized luciferase as a probe. From the bright-field images we obtained the local deflection angle to the vertical, A, local curvature, C, and the partial derivative of C with respect to time, [Formula: see text]. These were determined every 19.9 µm along the curvilinear length of the hypocotyl, at ~10 min intervals over a period of ~6 h after turning hypocotyls through 90° to the horizontal. Similarly from the luminescence images we measured the luminescence intensity of the convex and concave flanks of the hypocotyl as well as along the median of the hypocotyl, to determine differential expression of auxin-inducible IAA19. Comparison of these parameters as a function of time and curvilinear length shows that the gravitropic response is composed of three successive elements: the first and second curving responses and a decurving response (autostraightening). The maximum of the first curving response occurs when A is 76° along the entire length of the hypocotyl, suggesting that A is the sole determinant in this response; in contrast, the decurving response is a function of both A and C, as predicted by the newly-proposed graviproprioception model (Bastien et al., Proc Natl Acad Sci USA 110:755-760, 2013). Further, differential expression of IAA19, with higher expression in the convex flank, is observed at A = 44°, and follows the Sachs' sine law. This also suggests that IAA19 is not involved in the first curving response. In summary, the gravitropic response of Arabidopsis hypocotyls consists of multiple elements that are each determined by separate principles.

  2. Effects of caffeine on circadian phase, amplitude and period evaluated in cells in vitro and peripheral organs in vivo in PER2::LUCIFERASE mice

    Science.gov (United States)

    Narishige, Seira; Kuwahara, Mari; Shinozaki, Ayako; Okada, Satoshi; Ikeda, Yuko; Kamagata, Mayo; Tahara, Yu; Shibata, Shigenobu

    2014-01-01

    Background and Purpose Caffeine is one of the most commonly used psychoactive substances. Circadian rhythms consist of the main suprachiasmatic nucleus (SCN) clocks and peripheral clocks. Although caffeine lengthens circadian rhythms and modifies phase changes in SCN-operated rhythms, the effects on caffeine on the phase, period and amplitude of peripheral organ clocks are not known. In addition, the role of cAMP/Ca2+ signalling in effects of caffeine on rhythm has not been fully elucidated. Experimental Approach We examined whether chronic or transient application of caffeine affects circadian period/amplitude and phase by evaluating bioluminescence rhythm in PER2::LUCIFERASE knock-in mice. Circadian rhythms were monitored in vitro using fibroblasts and ex vivo and in vivo for monitoring of peripheral clocks. Key Results Chronic application of caffeine (0.1–10 mM) increased period and amplitude in vitro. Transient application of caffeine (10 mM) near the bottom of the decreasing phase of bioluminescence rhythm caused phase advance in vitro. Caffeine (0.1%) intake caused a phase delay under light–dark or constant dark conditions, suggesting a period-lengthening effect in vivo. Caffeine (20 mg·kg−1) at daytime or at late night-time caused phase advance or delay in bioluminescence rhythm in the liver and kidney respectively. The complicated roles of cAMP/Ca2+ signalling may be involved in the caffeine-induced increase of period and amplitude in vitro. Conclusions and Implications Caffeine affects circadian rhythm in mice by lengthening the period and causing a phase shift of peripheral clocks. These results suggest that caffeine intake with food/drink may help with food-induced resetting of peripheral circadian clocks. PMID:25160990

  3. DNA repair in human fibroblasts, as reflected by host-cell reactivation of a transfected UV-irradiated luciferase gene, is not related to donor age

    International Nuclear Information System (INIS)

    Merkle, Thomas J.; O'Brien, Katherine; Brooks, Philip J.; Tarone, Robert E.; Robbins, Jay H.

    2004-01-01

    The effect of donor age on the ability of mammalian cells to repair ultraviolet (UV)-induced DNA damage has been studied using several approaches, most recently via assays that measure the host-cell reactivation (HCR) of UV-irradiated reporter gene-containing plasmid vectors following their transfection into cells. Plasmid HCR assays indirectly quantify a cell line's ability to perform nucleotide excision repair (NER) by measuring the enzyme activity of the repaired reporter gene, e.g., chloramphenical acetyltransferase (cat) or luciferase (luc), and are useful in studies investigating whether increasing age may be a risk factor for the deficient repair of potentially cancer-causing, sunlight-induced, DNA lesions in skin cells. In our study, we quantified the DNA repair ability of cultured, nontransformed, human skin fibroblast lines through their HCR of a transfected UV-C-irradiated plasmid containing luc. HCR was measured at various times after transfection in five lines from normal donors of ages 21-96 years, and from one donor who had xeroderma pigmentosum (XP). The normal lines displayed increasing HCR at successive post-transfection time points and showed no significant correlation between HCR and donor age. The XP-A line, known to be markedly deficient in NER of UV-induced DNA damage, showed minimal evidence of HCR compared to the normal lines. To further assess potential variation in HCR with donor age, fibroblast lines from five old donors, ages 84-94 years, were compared with lines from five young donors, ages 17-26 years. While significant differences in HCR were found between some lines, no significant difference was found between the young and old age groups (P=0.44). Our study provides no indication that the higher incidence of skin cancer observed with increasing age is due to an age-related decrease in the ability to repair UV-induced DNA damage

  4. DNA repair in human fibroblasts, as reflected by host-cell reactivation of a transfected UV-irradiated luciferase gene, is not related to donor age

    Energy Technology Data Exchange (ETDEWEB)

    Merkle, Thomas J.; O' Brien, Katherine; Brooks, Philip J.; Tarone, Robert E.; Robbins, Jay H

    2004-10-04

    The effect of donor age on the ability of mammalian cells to repair ultraviolet (UV)-induced DNA damage has been studied using several approaches, most recently via assays that measure the host-cell reactivation (HCR) of UV-irradiated reporter gene-containing plasmid vectors following their transfection into cells. Plasmid HCR assays indirectly quantify a cell line's ability to perform nucleotide excision repair (NER) by measuring the enzyme activity of the repaired reporter gene, e.g., chloramphenical acetyltransferase (cat) or luciferase (luc), and are useful in studies investigating whether increasing age may be a risk factor for the deficient repair of potentially cancer-causing, sunlight-induced, DNA lesions in skin cells. In our study, we quantified the DNA repair ability of cultured, nontransformed, human skin fibroblast lines through their HCR of a transfected UV-C-irradiated plasmid containing luc. HCR was measured at various times after transfection in five lines from normal donors of ages 21-96 years, and from one donor who had xeroderma pigmentosum (XP). The normal lines displayed increasing HCR at successive post-transfection time points and showed no significant correlation between HCR and donor age. The XP-A line, known to be markedly deficient in NER of UV-induced DNA damage, showed minimal evidence of HCR compared to the normal lines. To further assess potential variation in HCR with donor age, fibroblast lines from five old donors, ages 84-94 years, were compared with lines from five young donors, ages 17-26 years. While significant differences in HCR were found between some lines, no significant difference was found between the young and old age groups (P=0.44). Our study provides no indication that the higher incidence of skin cancer observed with increasing age is due to an age-related decrease in the ability to repair UV-induced DNA damage.

  5. Kaempferol nanoparticles achieve strong and selective inhibition of ovarian cancer cell viability

    Science.gov (United States)

    Luo, Haitao; Jiang, Bingbing; Li, Bingyun; Li, Zhaoliang; Jiang, Bing-Hua; Chen, Yi Charlie

    2012-01-01

    Ovarian cancer is one of the leading causes of cancer death for women throughout the Western world. Kaempferol, a natural flavonoid, has shown promise in the chemoprevention of ovarian cancer. A common concern about using dietary supplements for chemoprevention is their bioavailability. Nanoparticles have shown promise in increasing the bioavailability of some chemicals. Here we developed five different types of nanoparticles incorporating kaempferol and tested their efficacy in the inhibition of viability of cancerous and normal ovarian cells. We found that positively charged nanoparticle formulations did not lead to a significant reduction in cancer cell viability, whereas nonionic polymeric nanoparticles resulted in enhanced reduction of cancer cell viability. Among the nonionic polymeric nanoparticles, poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) nanoparticles incorporating kaempferol led to significant reduction in cell viability of both cancerous and normal cells. Poly(DL-lactic acid-co-glycolic acid) (PLGA) nanoparticles incorporating kaempferol resulted in enhanced reduction of cancer cell viability together with no significant reduction in cell viability of normal cells compared with kaempferol alone. Therefore, both PEO-PPO-PEO and PLGA nanoparticle formulations were effective in reducing cancer cell viability, while PLGA nanoparticles incorporating kaempferol had selective toxicity against cancer cells and normal cells. A PLGA nanoparticle formulation could be advantageous in the prevention and treatment of ovarian cancers. On the other hand, PEO-PPO-PEO nanoparticles incorporating kaempferol were more effective inhibitors of cancer cells, but they also significantly reduced the viability of normal cells. PEO-PPO-PEO nanoparticles incorporating kaempferol may be suitable as a cancer-targeting strategy, which could limit the effects of the nanoparticles on normal cells while retaining their potency against cancer cells. We

  6. Assessment of the technical viability of reactor options for plutonium disposition

    International Nuclear Information System (INIS)

    Primm, R.T. III.

    1996-01-01

    Various reactor concepts for the disposition of surplus Pu have been proposed by reactor vendors; not all have attained the same level of technical viability. Studies were performed to differentiate between reactor concepts by devising a quantitative index for technical viability. For a quantitative assessment, three issues required resolution: the definition of a technical maturity scale, the treatment of ''subjective'' factors which cannot be easily represented in a quantitative format, and the protocol for producing a single technical viability figure-of-merit for each alternative. Alternatives involving the use of foreign facilities were found to be the most technically viable

  7. Study progress of cardiac MRI technology in assessment of myocardial viability after myocardial infarction

    International Nuclear Information System (INIS)

    Wang Jing; Zhang Hao

    2013-01-01

    Acute myocardial infarction (AMI) is one of the most common diseases that cause disability and death around the world. Correctly and effectively assessing the myocardial viability after myocardial infarction can reduce the disabled rate and mortality rate. At present, many methods could be used to assess myocardial viability. The cardiac magnetic resonance imaging (CMR) technology has a lot of advantages compared to other methods. In this paper, we reviewed the research progress of CMR in assessment of myocardial viability after myocardial infarction, and compared CMR with other technologies. (authors)

  8. Present trends in the detection of myocardial viability using nuclear cardiology tests

    International Nuclear Information System (INIS)

    Peix Gonzalez, Amalia; Garcia Barreto, David

    1999-01-01

    The myocardial viability diagnosis is important for those who will undergo myocardial revascularization whether by surgery or coronary angioplasty. Our purpose is to present some of the present trends in the detection of myocardial viability using nuclear cardiology tests. Emphasis is made on the estimation of radiopharmaceutical uptake and the use of vasodilators in perfusion scintigraphy mainly with technetium-labeled compounds. Also, the current possibilities for a myocardial metabolism study using single-photon emission-computed tomography as well as some clinical implications of myocardial viability are set forth

  9. Viability of Staphylococcus xylosus isolated from artisanal sausages for application as starter cultures in meat products.

    Science.gov (United States)

    Fiorentini, Angela Maria; Sawitzki, Maristela Cortez; Bertol, Teresinha Marisa; Sant'anna, Ernani S

    2009-01-01

    Viability of Staphylococcus xylosus isolated from artisanal sausages for application as starter cultures in meat products Viability of Staphylococcus xylosus strains AD1 and U5 isolated from natural fermented sausages was investigated as starter cultures in fermented sausages produced in the South Region of Brazil. The study demonstrated that the Staphylococcus xylosus strains AD1 and U5 showed significant growth during fermentation, stability over freeze-dried process, negative reaction for staphylococcal enterotoxins and viability for using as a single-strain culture or associated with lactic acid bacteria for production of fermented sausages.

  10. The viability of biological treatment at Ibi wastewater treatment station; Viabilidad del tratamiento biologico de aguas residuales

    Energy Technology Data Exchange (ETDEWEB)

    Flor Garcia, M.V.; Morenilla Martinez, J.J.; Ruiz Zapata, R.

    1996-06-01

    In this study, we have proved the viability of biological treatment of leaving waters from Ibi Wastewater Treatment Station, where water is subject to the action of coagulant agents, following a physical and chemical process. the experience was based on wastewater treatment by using activated sludge, at experimental scale in a pilot plant. During the experiments, we controlled the main parameters which indicate treatment success; namely, Suspended Solids (SS), pH, Biochemical Oxygen Demand (BOD) and Chemical Oxygen Demand (COD), in addition to other substances such as nutrients and toxicants, and inhibitors of bio metabolism. (Author) 6 refs.

  11. Rapid, portable and cost-effective yeast cell viability and concentration analysis using lensfree on-chip microscopy and machine learning

    KAUST Repository

    Feizi, Alborz

    2016-09-24

    Monitoring yeast cell viability and concentration is important in brewing, baking and biofuel production. However, existing methods of measuring viability and concentration are relatively bulky, tedious and expensive. Here we demonstrate a compact and cost-effective automatic yeast analysis platform (AYAP), which can rapidly measure cell concentration and viability. AYAP is based on digital in-line holography and on-chip microscopy and rapidly images a large field-of-view of 22.5 mm2. This lens-free microscope weighs 70 g and utilizes a partially-coherent illumination source and an opto-electronic image sensor chip. A touch-screen user interface based on a tablet-PC is developed to reconstruct the holographic shadows captured by the image sensor chip and use a support vector machine (SVM) model to automatically classify live and dead cells in a yeast sample stained with methylene blue. In order to quantify its accuracy, we varied the viability and concentration of the cells and compared AYAP\\'s performance with a fluorescence exclusion staining based gold-standard using regression analysis. The results agree very well with this gold-standard method and no significant difference was observed between the two methods within a concentration range of 1.4 × 105 to 1.4 × 106 cells per mL, providing a dynamic range suitable for various applications. This lensfree computational imaging technology that is coupled with machine learning algorithms would be useful for cost-effective and rapid quantification of cell viability and density even in field and resource-poor settings.

  12. Rapid, portable and cost-effective yeast cell viability and concentration analysis using lensfree on-chip microscopy and machine learning.

    Science.gov (United States)

    Feizi, Alborz; Zhang, Yibo; Greenbaum, Alon; Guziak, Alex; Luong, Michelle; Chan, Raymond Yan Lok; Berg, Brandon; Ozkan, Haydar; Luo, Wei; Wu, Michael; Wu, Yichen; Ozcan, Aydogan

    2016-11-01

    Monitoring yeast cell viability and concentration is important in brewing, baking and biofuel production. However, existing methods of measuring viability and concentration are relatively bulky, tedious and expensive. Here we demonstrate a compact and cost-effective automatic yeast analysis platform (AYAP), which can rapidly measure cell concentration and viability. AYAP is based on digital in-line holography and on-chip microscopy and rapidly images a large field-of-view of 22.5 mm 2 . This lens-free microscope weighs 70 g and utilizes a partially-coherent illumination source and an opto-electronic image sensor chip. A touch-screen user interface based on a tablet-PC is developed to reconstruct the holographic shadows captured by the image sensor chip and use a support vector machine (SVM) model to automatically classify live and dead cells in a yeast sample stained with methylene blue. In order to quantify its accuracy, we varied the viability and concentration of the cells and compared AYAP's performance with a fluorescence exclusion staining based gold-standard using regression analysis. The results agree very well with this gold-standard method and no significant difference was observed between the two methods within a concentration range of 1.4 × 10 5 to 1.4 × 10 6 cells per mL, providing a dynamic range suitable for various applications. This lensfree computational imaging technology that is coupled with machine learning algorithms would be useful for cost-effective and rapid quantification of cell viability and density even in field and resource-poor settings.

  13. A machine vision system for automated non-invasive assessment of cell viability via dark field microscopy, wavelet feature selection and classification

    Directory of Open Access Journals (Sweden)

    Friehs Karl

    2008-10-01

    Full Text Available Abstract Background Cell viability is one of the basic properties indicating the physiological state of the cell, thus, it has long been one of the major considerations in biotechnological applications. Conventional methods for extracting information about cell viability usually need reagents to be applied on the targeted cells. These reagent-based techniques are reliable and versatile, however, some of them might be invasive and even toxic to the target cells. In support of automated noninvasive assessment of cell viability, a machine vision system has been developed. Results This system is based on supervised learning technique. It learns from images of certain kinds of cell populations and trains some classifiers. These trained classifiers are then employed to evaluate the images of given cell populations obtained via dark field microscopy. Wavelet decomposition is performed on the cell images. Energy and entropy are computed for each wavelet subimage as features. A feature selection algorithm is implemented to achieve better performance. Correlation between the results from the machine vision system and commonly accepted gold standards becomes stronger if wavelet features are utilized. The best performance is achieved with a selected subset of wavelet features. Conclusion The machine vision system based on dark field microscopy in conjugation with supervised machine learning and wavelet feature selection automates the cell viability assessment, and yields comparable results to commonly accepted methods. Wavelet features are found to be suitable to describe the discriminative properties of the live and dead cells in viability classification. According to the analysis, live cells exhibit morphologically more details and are intracellularly more organized than dead ones, which display more homogeneous and diffuse gray values throughout the cells. Feature selection increases the system's performance. The reason lies in the fact that feature

  14. The viability of balancing wind generation with large scale energy storage

    International Nuclear Information System (INIS)

    Nyamdash, Batsaikhan; Denny, Eleanor; O'Malley, Mark

    2010-01-01

    This paper studies the impact of combining wind generation and dedicated large scale energy storage on the conventional thermal plant mix and the CO 2 emissions of a power system. Different strategies are proposed here in order to explore the best operational strategy for the wind and storage system in terms of its effect on the net load. Furthermore, the economic viability of combining wind and large scale storage is studied. The empirical application, using data for the Irish power system, shows that combined wind and storage reduces the participation of mid-merit plants and increases the participation of base-load plants. Moreover, storage negates some of the CO 2 emissions reduction of the wind generation. It was also found that the wind and storage output can significantly reduce the variability of the net load under certain operational strategies and the optimal strategy depends on the installed wind capacity. However, in the absence of any supporting mechanism none of the storage devices were economically viable when they were combined with the wind generation on the Irish power system. - Research Highlights: → Energy storage would displace the peaking and mid-merit plants generations by the base-load plants generations. Energy storage may negate the CO 2 emissions reduction that is due to the increased wind generations. →Energy storage reduces the variation of the net load. →Under certain market conditions, merchant type energy storage is not viable.

  15. Study of the economic viability of the innovative nuclear reactor SMART in Brazil

    International Nuclear Information System (INIS)

    Escanhoela, Cordelia Mara Fazzio; Lima, Ana Cecilia de Souza; Sabundjian, Gaianê

    2017-01-01

    The main objective of this study is to evaluate the economic viability of the installation and operation of the innovative System - Integrated Modular Advanced Reactor (SMART) in Brazil. SMART, developed by the Korea Atomic Energy Research Institute (KAERI), is a small and modular Power Water Reactor (PWR), presents electric power of 100 MW and thermal power of 330 MW; it has a passive safety system and integral refrigeration configuration, characteristics that, allied with modularization, simplification and technological improvements, give SMART greater reliability and economy when compared to conventional reactors. SMART presents, in addition to electricity production, the functions of seawater desalination and district heat generation. The research is based on projections of energy demand in the medium and long term with emphasis on electricity and search for the reduction of greenhouse gases. These previsions indicate the need for energy expansion and diversification of the current sources in Brazil, predominantly water sources. The methodology used is based on the cost of electric generation, production capacity and construction time of SMART, adopting the investment model similar to the Angra 3 plant and the use of mirrored costs between the plants. The feasibility of the project was evaluated through the financial criteria: Internal Rate of Return (IRR), Net Present Value (NPV) and Weighted Average Cost of Capital (WACC), whose revenue should be generated through a tariff passed on to the consumer. (author)

  16. Study of the economic viability of the innovative nuclear reactor SMART in Brazil

    Energy Technology Data Exchange (ETDEWEB)

    Escanhoela, Cordelia Mara Fazzio; Lima, Ana Cecilia de Souza; Sabundjian, Gaianê, E-mail: liafazzio@hotmail.com, E-mail: aclima@ipen.br, E-mail: gdjian@ipen.br [Centro de Engenharia Nuclear - CEN, Instituto de Pesquisas Energéticas e Nucleares (IPEN/CNEN-SP), São Paulo, SP (Brazil)

    2017-07-01

    The main objective of this study is to evaluate the economic viability of the installation and operation of the innovative System - Integrated Modular Advanced Reactor (SMART) in Brazil. SMART, developed by the Korea Atomic Energy Research Institute (KAERI), is a small and modular Power Water Reactor (PWR), presents electric power of 100 MW and thermal power of 330 MW; it has a passive safety system and integral refrigeration configuration, characteristics that, allied with modularization, simplification and technological improvements, give SMART greater reliability and economy when compared to conventional reactors. SMART presents, in addition to electricity production, the functions of seawater desalination and district heat generation. The research is based on projections of energy demand in the medium and long term with emphasis on electricity and search for the reduction of greenhouse gases. These previsions indicate the need for energy expansion and diversification of the current sources in Brazil, predominantly water sources. The methodology used is based on the cost of electric generation, production capacity and construction time of SMART, adopting the investment model similar to the Angra 3 plant and the use of mirrored costs between the plants. The feasibility of the project was evaluated through the financial criteria: Internal Rate of Return (IRR), Net Present Value (NPV) and Weighted Average Cost of Capital (WACC), whose revenue should be generated through a tariff passed on to the consumer. (author)

  17. The effect of CO{sub 2} laser beam welded AISI 316L austenitic stainless steel on the viability of fibroblast cells, in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Köse, Ceyhun, E-mail: ceyhun.kose@gop.edu.tr [Faculty of Natural Sciences and Engineering, Department of Mechanical Engineering, Gaziosmanpaşa University, Tokat (Turkey); Kaçar, Ramazan, E-mail: rkacar@karabuk.edu.tr [Faculty of Technology Department of Manufacturing Engineering, Karabuk University, Karabuk 78050 (Turkey); Zorba, Aslı Pınar, E-mail: aslipinarzorba@gmail.com [Graduate School of Natural and Applied Sciences, Department of Bioengineering Cell Culture and Tissue Engineering, Yıldız Technical University, Istanbul (Turkey); Bağırova, Melahat, E-mail: mbagir@yildiz.edu.tr [Department of Bioengineering Cell Culture and Tissue Engineering, Yıldız Technical University, Istanbul (Turkey); Allahverdiyev, Adil M., E-mail: adil@yildiz.edu.tr [Department of Bioengineering Cell Culture and Tissue Engineering, Yıldız Technical University, Istanbul (Turkey)

    2016-03-01

    It has been determined by the literature research that there is no clinical study on the in vivo and in vitro interaction of the cells with the laser beam welded joints of AISI 316L biomaterial. It is used as a prosthesis and implant material and that has adequate mechanical properties and corrosion resistance characteristics. Therefore, the interaction of the CO{sub 2} laser beam welded samples and samples of the base metal of AISI 316L austenitic stainless steel with L929 fibroblast cells as an element of connective tissue under in vitro conditions has been studied. To study the effect of the base metal and the laser welded test specimens on the viability of the fibroblast cells that act as an element of connective tissues in the body, they were kept in DMEMF-12 medium for 7, 14, 28 days and 18 months. The viability study was experimentally studied using the MTT method for 7, 14, 28 days. In addition, the direct interaction of the fibroblast cells seeded on 6 different plates with the samples was examined with an inverted microscope. The MTT cell viability experiment was repeated on the cells that were in contact with the samples. The statistical relationship was analyzed using a Tukey test for the variance with the GraphPad statistics software. The data regarding metallic ion release were identified with the ICP-MS method after the laser welded and main material samples were kept in cell culture medium for 18 months. The cell viability of the laser welded sample has been detected to be higher than that of the base metal and the control based on 7th day data. However, the laser welded sample's viability of the fibroblast cells has diminished by time during the test period of 14 and 28 days and base metal shows better viability when compared to the laser welded samples. On the other hand, the base metal and the laser welded sample show better cell viability effect when compared to the control group. According to the ICP-MS results of the main material and

  18. NPP Cernavoda unit 2 economic viability: The challenge for an advanced financing scheme

    International Nuclear Information System (INIS)

    Condu, M.; Popescu, D.

    1999-01-01

    The completion of Cernavoda Unit-2 being no doubt, the viability of the project was analyzed and strengthened. Economic justification of the decision to complete Unit-2 is described, including assessment of its safety design guides, design requirements and engineering design solutions in the light of changes in codes, guideline, standards and actual regulatory requirements. Cost-benefit analysis demonstrates the economic viability of the project

  19. Determination of Complement-Mediated Killing of Bacteria by Viability Staining and Bioluminescence

    OpenAIRE

    Virta, Marko; Lineri, Sanna; Kankaanpää, Pasi; Karp, Matti; Peltonen, Karita; Nuutila, Jari; Lilius, Esa-Matti

    1998-01-01

    Complement-mediated killing of bacteria was monitored by flow cytometric, luminometric, and conventional plate counting methods. A flow cytometric determination of bacterial viability was carried out by using dual staining with a LIVE/DEAD BacLight bacterial viability kit. In addition to the viable cell population, several other populations emerged in the fluorescence histogram, and there was a dramatic decrease in the total cell count in the light-scattering histogram in the course of the co...

  20. The Effect of Method and Germination Paper Substrate on Viability of Eucalyptus Pellita F. Mull Seed

    OpenAIRE

    Yuniarti, Naning; Megawati, Megawati; Leksono, Budi

    2017-01-01

    Improper seed handling of Eucalyptus pellita will reduce seed quality, so as to improve the viability of the seed proper handling techniques are needed. To investigate the seed germination potency as a result of seed handling germination seed tests are needed. The purpose of this study was to examine the effect of germination method and paper substrate on the viability of E. pellita seeds. Seeds used in this study were from seedling seed orchard in South Sumatra, South Kalimantan, and Riau. ...

  1. Hospital board effectiveness: relationships between board training and hospital financial viability.

    Science.gov (United States)

    Molinari, C; Morlock, L; Alexander, J; Lyles, C A

    1992-01-01

    This study examined whether hospital governing boards that invest in board education and training are more informed and effective decision-making bodies. Measures of hospital financial viability (i.e., selected financial ratios and outcomes) are used as indicators of hospital board effectiveness. Board participation in educational programs was significantly associated with improved profitability, liquidity, and occupancy levels, suggesting that investment in the education of directors is likely to enhance hospital viability and thus increase board effectiveness.

  2. Viability of Lactobacillus acidophilus and Lactobacillus casei in fermented milk products during refrigerated storage.

    Science.gov (United States)

    Nighswonger, B D; Brashears, M M; Gilliland, S E

    1996-02-01

    The viability was investigated of five strains of Lactobacillus acidophilus and one strain of Lactobacillus casei that were added as adjuncts to yogurt and cultured buttermilk during 28 d of refrigerated storage at 5 to 7 degrees C. A modification of LBS (Lactobacillus selection) agar was used for the enumeration of L. acidophilus and L. casei. The medium allowed the colony formation of the adjunct bacteria while preventing colony formation of the traditional yogurt or buttermilk starter cultures. At each sampling period, colonies from the selective agar medium were isolated for confirmation of identity to confirm that only L. acidophilus and L. casei were enumerated, that their characteristics did not change during storage, or both. The strains of L. acidophilus varied in both cultured products. In buttermilk, L. acidophilus MUH-41, O-16, and L-1 exhibited no significant loss in viability, but strains 43121 and La-5 did. No significant loss in viability of L. acidophilus MUH-41 and L-1 occurred in yogurt prepared using culture CM2; however, strains 43121, O-16, and La-5 lost viability. In the yogurt prepared using culture YC-4, L. acidophilus 43121 exhibited no significant loss in viability, but MUH-41, O-16, L-1, and La-5 did. There was no loss in viability of L. casei GG during storage of any of the cultured products.

  3. An updated methodology to review developing-country vaccine manufacturer viability.

    Science.gov (United States)

    Luter, Nicholas; Kumar, Ritu; Hozumi, Dai; Lorenson, Tina; Larsen, Shannon; Gowda, Bhavya; Batson, Amie

    2017-07-05

    In 1997, Milstien, Batson, and Meaney published "A Systematic Method for Evaluating the Potential Viability of Local Vaccine Producers." The paper identified characteristics of successful vaccine manufacturers and developed a viability framework to evaluate their performance. This paper revisits the original study after two decades to determine the ability of the framework to predict manufacturer success. By reconstructing much of the original dataset and conducting in-depth interviews, the authors developed informed views on the continued viability of manufacturers in low- and middle-income country markets. Considering the marked changes in the market and technology landscape since 1997, the authors find the viability framework to be predictive and a useful lens through which to evaluate manufacturer success or failure. Of particular interest is how incumbent and potentially new developing-country vaccine manufacturers enter and sustain production in competitive international markets and how they integrate (or fail to integrate) new technology into the production process. Ultimately, most manufacturers will need to meet global quality standards to be viable. As governments and donors consider investments in vaccine producers, the updated viability factors will be a useful tool in evaluating the prospects of manufacturers over the mid to long term. The paper emphasizes that while up-front investments are important, other critical factors-including investments in a national regulatory authority, manufacturer independence, and ability to adapt and adopt new technology-are necessary to ensure viability. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  4. Monitoring change in refractive index of cytosol of animal cells on affinity surface under osmotic stimulus for label-free measurement of viability.

    Science.gov (United States)

    Park, Jina; Jin, Sung Il; Kim, Hyung Min; Ahn, Junhyoung; Kim, Yeon-Gu; Lee, Eun Gyo; Kim, Min-Gon; Shin, Yong-Beom

    2015-02-15

    We demonstrated that a metal-clad waveguide (MCW)-based biosensor can be applied to label-free measurements of viability of adherent animal cells with osmotic stimulation in real time. After Chinese hamster ovary (CHO) and human embryonic kidney cell 293 (HEK293) cells were attached to a Concanavalin A (Con A)-modified sensor surface, the magnitudes of cell responses to non-isotonic stimulation were compared between live and dead cells. The live cells exhibited a change in the refractive index (RI) of the cytosol caused by a redistribution of water through the cell membrane, which was induced by the osmotic stimulus, but the dead cells did not. Moreover, the normalized change in the RI measured via the MCW sensor was linearly proportional to the viability of attached cells and the resolution in monitoring cell viability was about 0.079%. Therefore, the viability of attached animal cells can be measured without labels by observing the relative differences in the RI of cytosol in isotonic and non-isotonic buffers. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Commercial viability of hybrid vehicles : best household use and cross national considerations.

    Energy Technology Data Exchange (ETDEWEB)

    Santini, D. J.; Vyas, A. D.

    1999-07-16

    Japanese automakers have introduced hybrid passenger cars in Japan and will soon do so in the US. In this paper, we report how we used early computer simulation model results to compare the commercial viability of a hypothetical near-term (next decade) hybrid mid-size passenger car configuration under varying fuel price and driving patterns. The fuel prices and driving patterns evaluated are designed to span likely values for major OECD nations. Two types of models are used. One allows the ''design'' of a hybrid to a specified set of performance requirements and the prediction of fuel economy under a number of possible driving patterns (called driving cycles). Another provides an estimate of the incremental cost of the hybrid in comparison to a comparably performing conventional vehicle. In this paper, the models are applied to predict the NPV cost of conventional gasoline-fueled vehicles vs. parallel hybrid vehicles. The parallel hybrids are assumed to (1) be produced at high volume, (2) use nickel metal hydride battery packs, and (3) have high-strength steel bodies. The conventional vehicle also is assumed to have a high-strength steel body. The simulated vehicles are held constant in many respects, including 0-60 time, engine type, aerodynamic drag coefficient, tire rolling resistance, and frontal area. The hybrids analyzed use the minimum size battery pack and motor to meet specified 0-60 times. A key characteristic affecting commercial viability is noted and quantified: that hybrids achieve the most pronounced fuel economy increase (best use) in slow, average-speed, stop-and-go driving, but when households consistently drive these vehicles under these conditions, they tend to travel fewer miles than average vehicles. We find that hours driven is a more valuable measure than miles. Estimates are developed concerning hours of use of household vehicles versus driving cycle, and the pattern of minimum NPV incremental cost (or benefit) of

  6. 99mTc-GHA: A promising agent for assessing tumour viability

    International Nuclear Information System (INIS)

    Choudhury, P.S.; Gupta, A.; Sharma, P.K.; Bhatia, S.; Nambiar, U.; Jena, A.

    2004-01-01

    Full text: Biochemical investigations have a major role to play in the management of primary malignancies. Major biochemical changes occur during cancerous transformation including changes in the energy metabolism of the cell. Changes can take place in terms of utilization of glucose and other substrates, protein synthesis and expression of antigens and receptors. Changes also take place in disruption of transport mechanism across cell membranes and other physiological boundaries like blood brain barrier. Anatomical imaging is aimed at the diagnosis of tumours whereas biochemical imaging is better suited for tissue characterization. We report here the utility of 99mTc GHA as a radiopharmaceutical for assessing tumour viability. Evaluation was done for 44 patients who had undergone 72 studies based on objective parameters like histopathological correlation and serial clinical and other radiological evaluation in long-term follow-up. All the evaluated patients had undergone magnetic resonance imaging (MRI) of the local site within one week of 99m GHA study and had some sort of morphological abnormality and suspicious contrast behaviour. 41 patients had cerebral lesions (37 primary brain tumours during initial diagnosis, evaluation of residual or recurrent disease and 4 patients of brain metastasis) and 3 patients had Ewing's Sarcoma evaluated during the course of treatment. 29 patients of suspected brain tumour (21 primary and 8 residual / recurrence) had surgical decompression along with 2/4 in metastatic brain lesions. Out of the 29 patients who underwent surgery, 28 patients had histopathology positive for malignancy. 27 patients showed abnormal GHA concentration corresponding to the morphological abnormality (1 false negative). One patient of pseudo-tumour also showed avid tracer concentration (false positive). 2/4 patients of solitary metastatic foci in brain, with positive GHA scan, underwent surgery and showed positive histopathology. 2/4 patients treated by

  7. Influence of different grained powders and pellets made of Niobium and Ti-42Nb on human cell viability

    Energy Technology Data Exchange (ETDEWEB)

    Markhoff, Jana, E-mail: markhoffj@gmail.com [University Medicine Rostock, Department of Orthopedics, Biomechanics and Implant Technology Laboratory, Doberaner Strasse 142, 18057 Rostock (Germany); Weinmann, Markus [H.C. Starck Tantalum and Niobium GmbH, Im Schleeke 78-91, 38642 Goslar (Germany); Schulze, Christian; Bader, Rainer [University Medicine Rostock, Department of Orthopedics, Biomechanics and Implant Technology Laboratory, Doberaner Strasse 142, 18057 Rostock (Germany)

    2017-04-01

    Nowadays, biomaterials can be used to maintain or replace several functions of the human body if necessary. Titanium and its alloys, i.e. Ti6Al4V are the most common materials (70 to 80%) used for structural orthopedic implants due to their unique combination of good mechanical properties, corrosion resistance and biocompatibility. Addition of β-stabilizers, e.g. niobium, can improve the mechanical properties of such titanium alloys further, simultaneously offering excellent biocompatibility. In this in vitro study, human osteoblasts and fibroblasts were cultured on different niobium specimens (Nb Amperit, Nb Ampertec), Nb sheets and Ti-42Nb (sintered and 3D-printed by selective laser melting, SLM) and compared with forged Ti6Al4V specimens. Furthermore, human osteoblasts were incubated with particulates of the Nb and Ti-42Nb specimens in three concentrations over four and seven days to imitate influence of wear debris. Thereby, the specimens with the roughest surfaces, i.e. Ti-42Nb and Nb Ampertec, revealed excellent and similar results for both cell types concerning cell viability and collagen synthesis superior to forged Ti6Al4V. Examinations with particulate debris disclosed a dose-dependent influence of all powders with Nb Ampertec showing the highest decrease of cell viability and collagen synthesis. Furthermore, interleukin synthesis was only slightly increased for all powders. In summary, Nb Ampertec (sintered Nb) and Ti-42Nb materials seem to be promising alternatives for medical applications compared to common materials like forged or melted Ti6Al4V. - Highlights: • Titanium and its alloys most common materials used for structural orthopedic implants • Addition of β-stabilizers to improve mechanical properties • Roughest surfaces, Nb ampertec and Ti-42Nb, with excellent results concerning cell viability and collagen synthesis • No cell-specific differences between human osteoblasts and fibroblasts • Niobium based powders with dose- and partly

  8. Technical and economic viability of electric power plants on the basis of renewable energy resources regarding hierarchical structure

    Directory of Open Access Journals (Sweden)

    Balzannikov Mikhail

    2017-01-01

    Full Text Available The article deals with power stations working on the basis of non-renewable energy resources and finite resources which will inevitably come to depletion in the future. These installations produce considerable negative impact on the environment, including air pollution. It is noted that considerable amounts of emissions of harmful substances accounts for the share of small thermal installations which aren’t always considered in calculations of pollution. The author specifies that emission reduction of harmful substances should be achieved due to wider use of environmentally friendly renewable energy resources. It is recommended to use hierarchical structure with the priority of ecological and social conditions of the region for technical and economic viability of consumers’ power supply systems and installations, based on renewable energy resources use. At the same time the author suggests considering federal, regional and object levels of viability. It is recommended to consider the main stages of lifecycle of an object for object level: designing, construction, operation, reconstruction of an object and its preservation. The author shows the example of calculation of power plant efficiency, based on renewable energy resources during its reconstruction, followed by power generation increase.

  9. Usefulness of dynamic contrast-enhanced MRI in the evaluation of the viability of acute scaphoid fracture

    Energy Technology Data Exchange (ETDEWEB)

    Larribe, Maud [Hopital La Conception, Service d' imagerie medicale, Marseille (France); Hopital Sainte Marguerite, Service d' imagerie medicale, Marseille (France); Gay, Andre [Hopital La Conception, Service de chirurgie de la main, Marseille (France); Freire, Veronique [Centre hospitalier de l' Universite de Montreal, Department of Radiology, Notre-Dame Hospital, Montreal, QC (Canada); Bouvier, Corinne [Hopital La Timone, Service d' anatomopathologie, Marseille (France); Chagnaud, Christophe; Souteyrand, Philippe [Hopital La Conception, Service d' imagerie medicale, Marseille (France)

    2014-12-15

    To evaluate the usefulness of dynamic gadolinium-enhanced magnetic resonance imaging (MRI) for assessing the viability of the proximal pole of the scaphoid in patients with acute scaphoid fractures. Eighteen consecutive patients with acute scaphoid fracture who underwent dynamic gadolinium-enhanced MRI 7 days or less before surgery were prospectively included between August 2011 and December 2012. All patients underwent MR imaging with unenhanced images, enhanced images, and dynamic enhanced images. A radiologist first classified the MRI results as necrotic or viable based on T1- and T2-weighted images only, followed by a second blinded interpretation, this time including analysis of pre- and post-gadolinium administration images and a third blinded interpretation based on the time-intensity curve of the dynamic enhanced study. The standard of reference was the histologic assessment of a cylindrical specimen of the proximal pole obtained during surgery in all patients. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated for unenhanced, enhanced, and dynamic gadolinium-enhanced MRI studies. The sensitivity, specificity, PPV, and NPV were 67, 67, 50, and 80 % for unenhanced images, 83, 100, 100, and 92 for enhanced images, and 83, 92, 83, and 92 for dynamic contrast-enhanced images. Our data are consistent with previously reported data supporting contrast-enhanced MRI for assessment of viability, and showing that dynamic imaging with time-intensity curve analysis does not provide additional predictive value over standard delayed enhanced imaging for acute scaphoid fracture. (orig.)

  10. Usefulness of dynamic contrast-enhanced MRI in the evaluation of the viability of acute scaphoid fracture

    International Nuclear Information System (INIS)

    Larribe, Maud; Gay, Andre; Freire, Veronique; Bouvier, Corinne; Chagnaud, Christophe; Souteyrand, Philippe

    2014-01-01

    To evaluate the usefulness of dynamic gadolinium-enhanced magnetic resonance imaging (MRI) for assessing the viability of the proximal pole of the scaphoid in patients with acute scaphoid fractures. Eighteen consecutive patients with acute scaphoid fracture who underwent dynamic gadolinium-enhanced MRI 7 days or less before surgery were prospectively included between August 2011 and December 2012. All patients underwent MR imaging with unenhanced images, enhanced images, and dynamic enhanced images. A radiologist first classified the MRI results as necrotic or viable based on T1- and T2-weighted images only, followed by a second blinded interpretation, this time including analysis of pre- and post-gadolinium administration images and a third blinded interpretation based on the time-intensity curve of the dynamic enhanced study. The standard of reference was the histologic assessment of a cylindrical specimen of the proximal pole obtained during surgery in all patients. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated for unenhanced, enhanced, and dynamic gadolinium-enhanced MRI studies. The sensitivity, specificity, PPV, and NPV were 67, 67, 50, and 80 % for unenhanced images, 83, 100, 100, and 92 for enhanced images, and 83, 92, 83, and 92 for dynamic contrast-enhanced images. Our data are consistent with previously reported data supporting contrast-enhanced MRI for assessment of viability, and showing that dynamic imaging with time-intensity curve analysis does not provide additional predictive value over standard delayed enhanced imaging for acute scaphoid fracture. (orig.)

  11. Fenton mediated ultrasonic disintegration of sludge biomass: Biodegradability studies, energetic assessment, and its economic viability.

    Science.gov (United States)

    Kavitha, S; Rajesh Banu, J; IvinShaju, C D; Kaliappan, S; Yeom, Ick Tae

    2016-12-01

    Mechanical disintegration of sludge through ultrasonication demands high energy and cost. Therefore, in the present study, a comprehensive investigation was performed to analyze the potential of a novel method, fenton mediated sonic disintegration (FSD). In FSD process, extracellular polymeric substance (EPS) of sludge was first removed via fenton treatment. It was subsequently disintegrated via ultrasonication. Energetic assessment and economic analysis were then performed using net energy and cost gain (spent) as key factor to evaluate the practical viability of the FSD process. FSD was found to be superior over sonic disintegration based on its higher sludge solubilization (34.4% vs. 23.2%) and methane production potential (0.3gCOD/gCOD vs. 0.2gCOD/gCOD). Both energy analysis and cost assessment of the present study revealed that FSD could reduce the energy demand of ultrasonication considerably with a positive net profit of about 44.93USD/Ton of sludge. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Cell viability and MRI performance of highly efficient polyol-coated magnetic nanoparticles

    International Nuclear Information System (INIS)

    Arteaga-Cardona, Fernando; Gutiérrez-García, Eric; Hidalgo-Tobón, Silvia; López-Vasquez, Ciro; Brito-Barrera, Yazmín A.; Flores-Tochihuitl, Julia; Angulo-Molina, Aracely; Reyes-Leyva, Julio R.; González-Rodríguez, Roberto; Coffer, Jeffery L.; Pal, Umapada

    2016-01-01

    This work aimed at determining conditions that would allow us to control the size of the NPs and create a system with characteristics apt for biomedical applications. We describe a comprehensive study on the synthesis and physical characterization of two highly sensitive sets of triethylene glycol (TREG) and polyethylene glycol (PEG)-coated superparamagnetic iron oxide nanoparticles (SPIONs) to be evaluated for use as magnetic resonance (MR) contrast agents. The ferrofluids demonstrated excellent colloidal stability in deionized water at pH 7.0 as indicated by dynamic light scattering (DLS) data. The magnetic relaxivities, r_2, were measured on a 1.5 T clinical MRI instrument. Values in the range from 205 to 257 mM"−"1 s"−"1 were obtained, varying proportionally to the SPIONs’ sizes and coating nature. Further in vitro cell viability tests and in vivo biodistribution analyses of the intravenously administered nanoparticles showed that the prepared systems have good biocompatibility and migrate to several organs, mainly the meninges, spleen, and liver. Based on these results, our findings demonstrated the potential utility of these nanosystems as clinical contrast agents for MR imaging.

  13. Viability study for the implantation of an incineration unit for low level radioactive wastes

    International Nuclear Information System (INIS)

    Andrade, Andre Wagner Oliani

    1995-01-01

    Incineration have been a world-wide accepted volume reduction technique for combustible materials due to its high efficiency and excellent results. This technique is used since the last century as an alternative to reduce cities garbage and during the last four decades for the hazardous wastes. The nuclear industry is also involved in this technique development related to the low level radioactive waste management. There are different types of incineration installations and the definition of the right system is based on a criterious survey of its main characteristics, related to the rad wastes as well technical, economical and burocratic parameters. After the autonomous Brazilian nuclear programme development and the onlook of the future intensive nuclear energy uses, a radwaste generation increase is expected. One of the installations where these radwastes volumes are awaited to be high is the Experimental Center of ARAMAR (CEA). Nuclear reactors for propulsion and power generation have been developed in CEA beyond other nuclear combustible cycle activities. In this panorama it is important to evaluate the incineration role in CEA installations, as a volume reduction technique for an appropriate radioactive wastes management implementation. In this work main aspects related to the low level radwaste incineration systems were up rised. This information are important to a coherent viability study and also to give a clear and impartial about a topic that is still non discussed in the national scenery. (author)

  14. Extracellular vesicle associated long non-coding RNAs functionally enhance cell viability

    Directory of Open Access Journals (Sweden)

    Chris Hewson

    2016-10-01

    Full Text Available Cells communicate with one another to create microenvironments and share resources. One avenue by which cells communicate is through the action of exosomes. Exosomes are extracellular vesicles that are released by one cell and taken up by neighbouring cells. But how exosomes instigate communication between cells has remained largely unknown. We present evidence here that particular long non-coding RNA molecules are preferentially packaged into exosomes. We also find that a specific class of these exosome associated non-coding RNAs functionally modulate cell viability by direct interactions with l-lactate dehydrogenase B (LDHB, high-mobility group protein 17 (HMG-17, and CSF2RB, proteins involved in metabolism, nucleosomal architecture and cell signalling respectively. Knowledge of this endogenous cell to cell pathway, those proteins interacting with exosome associated non-coding transcripts and their interacting domains, could lead to a better understanding of not only cell to cell interactions but also the development of exosome targeted approaches in patient specific cell-based therapies. Keywords: Non-coding RNA, Extracellular RNA, Exosomes, Retroelement, Pseudogene

  15. Economic viability of geothermal energy usage in comparison to renewable and conventional energy systems

    International Nuclear Information System (INIS)

    Schaumann, G.

    2002-01-01

    This comprehensive lecture given by Prof. Dr. Gunter Schaumann in Bad Duerkheim, Germany, discusses the use of geothermal energy in relationship to other forms of renewable energy sources and conventional energy technologies used to provide heat, power and motive force. The characteristics of geothermal energy from various sources and examples of its possible use are discussed. In particular, the paper deals with deep geothermal energy, which can provide heating energy for district heating schemes, if necessary with the help of heat pumps. The prospects of such a use of geothermal energy in the next 50 years in various suitable regions in Germany is discussed and the associated prerequisites are listed. The present situation concerning the use of geothermal energy in Germany is examined. An example of a geothermal heating power station that also features a gas-fired combined heat and power installation, a heat pump and a peak-load boiler is given. Also, the generation of electrical power using the Organic Rankine Cycle is discussed. The factors influencing the economic viability of geothermal power stations are discussed in detail and the resulting energy prices are compared with conventional plants. The paper gives details of the calculation of investment and energy costs for heat and power generation and presents figures based on exemplary installations

  16. Economic viability of potted chrysanthemums production in Atibaia, São Paulo state

    Directory of Open Access Journals (Sweden)

    Caio Shigueaki Shiroto

    2016-08-01

    Full Text Available The segment of flower production in Brazil has shown remarkable development in recent years. The chrysanthemum is a product of extensive sales throughout Brazil and the diversity of types and colors, resistance to transport, excellent durability, and its easy adaptation to different regions make it as one of the main products in the various markets. This study aimed to evaluate the cost and economic viability of commercial production of potted chrysanthemums in the Atibaia, São Paulo State. For the total cost of production (6,413 vases/month expenses cuttings accounted for 36.4% of inputs and 26.4% of the EOC (effective operational cost, followed by labor, with 16% of the TOC (total operational cost achieving a profitability index 27.7%. It was found, based on cash flow, an IRR (internal rate of return of 10.27% IRR (internal rate of return already for the 6th productive year, showing attractive results for this segment considering the improving producer profitability is proportional to better production indicators. Note that to get a higher return activity, more efficient managements are required, resulting in lower losses and higher operating earnings, being necessary to take into account the cost management and production system are also essential to success in cultivation.

  17. The effects of beryllium metal particles on the viability and function of cultured rat alveolar macrophages

    International Nuclear Information System (INIS)

    Finch, G.L.; Lowther, W.T.; Hoover, M.D.; Brooks, A.L.

    1988-01-01

    Rat pulmonary alveolar macrophages (PAM) were exposed in vitro to beryllium metal particles. The particles used were relatively large (Be-II) and small (Be-V) size fractions of beryllium metal obtained from an aerosol cyclone, and a beryllium metal aerosol generated by laser vaporization of beryllium metal in an argon atmosphere (Be-L). Glass beads (GB) were used as a negative control particle. The endpoints examined included cell killing (trypan blue dye exclusion) and phagocytic ability (sheep red blood cell uptake). Phagocytic ability was inhibited by beryllium particles at concentrations that did not cause appreciable cell killing. Results based on the mass concentration of particles in culture medium were transformed by the amount of specific surface area of the particles to permit expression of toxicity on the basis of amount of surface area of particles per unit volume of culture medium. On a mass concentration basis, the order of cytotoxicity was Be-L > Be-V ∼ Be-II > GB; for inhibition of phagacytosis, the cytotoxicity order was Be-L ∼ Be-V > Be-II > GB. On a surface area concentration basis, the order of toxicity for viability was altered to Be-II > Be-L ∼ Be-V (with GB indeterminant) and to Be-V > Be-II ∼ Be-L > GB for inhibition of phagocytosis. We conclude that there are factors in addition to specific surface area that influence the expression of toxic effects in cultured PAM. (author)

  18. Cell viability and MRI performance of highly efficient polyol-coated magnetic nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Arteaga-Cardona, Fernando [Universidad de las Américas de Puebla, Departamento de Ciencias Químico-Biológicas (Mexico); Gutiérrez-García, Eric [Instituto Literario, Universidad Autónoma del Estado de México (Mexico); Hidalgo-Tobón, Silvia, E-mail: shid@xanum.uam.mx [Universidad Autónoma Metropolitana, Departamento de Física (Mexico); López-Vasquez, Ciro; Brito-Barrera, Yazmín A. [Universidad de las Américas de Puebla, Departamento de Ciencias Químico-Biológicas (Mexico); Flores-Tochihuitl, Julia [Benemérita Universidad Autónoma de Puebla, Facultad de Estomatología (Mexico); Angulo-Molina, Aracely [Universidad de Sonora, Departamento de Ciencias Químico-Biológicas (Mexico); Reyes-Leyva, Julio R. [Instituto Mexicano del Seguro Social, Centro de Investigación Biomédica de Oriente (CIBIOR) (Mexico); González-Rodríguez, Roberto; Coffer, Jeffery L. [Texas Christian University, Department of Chemistry (United States); Pal, Umapada [Benemérita Universidad Autónoma de Puebla, Apdo, Instituto de Física (Mexico); and others

    2016-11-15

    This work aimed at determining conditions that would allow us to control the size of the NPs and create a system with characteristics apt for biomedical applications. We describe a comprehensive study on the synthesis and physical characterization of two highly sensitive sets of triethylene glycol (TREG) and polyethylene glycol (PEG)-coated superparamagnetic iron oxide nanoparticles (SPIONs) to be evaluated for use as magnetic resonance (MR) contrast agents. The ferrofluids demonstrated excellent colloidal stability in deionized water at pH 7.0 as indicated by dynamic light scattering (DLS) data. The magnetic relaxivities, r{sub 2}, were measured on a 1.5 T clinical MRI instrument. Values in the range from 205 to 257 mM{sup −1} s{sup −1} were obtained, varying proportionally to the SPIONs’ sizes and coating nature. Further in vitro cell viability tests and in vivo biodistribution analyses of the intravenously administered nanoparticles showed that the prepared systems have good biocompatibility and migrate to several organs, mainly the meninges, spleen, and liver. Based on these results, our findings demonstrated the potential utility of these nanosystems as clinical contrast agents for MR imaging.

  19. Examination of viability and quality of ovarian tissue after cryopreservation using simple laboratory methods in ewe

    Directory of Open Access Journals (Sweden)

    Guerin Jean F

    2011-06-01

    Full Text Available Abstract Background The objective of the present study is to assess viability tests and to evaluate follicle ovarian tissue quality after freezing-thawing procedures. Methods Ewe's ovaries were harvested at the slaughterhouse, after dissection each ovarian specimen was divided into two groups: fresh tissue (control group and frozen tissue. In the first part of the study, the follicles viability was assessed by trypan blue staining, calcein AM/ethidium homodimer-1 staining (LIVE/DEAD viability/cytotoxicity kit, Molecular Probes and morphology in the two groups. In the second part of the study the quality of the whole ovarian tissue was evaluated by the quantification of the release of lactate dehydrogenase measurement (Cytotoxicity Detection kit ROCHE, DNA fragmentation by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL in primordial and primary follicles (ApopDETEK Kit system Enzo and morphology in the two groups. 100 Follicles (primordial and primary were counted on both fresh and frozen hemiovary to assess this various tests. Results Ovarian follicle viability assessment was similar using trypan blue or calcein/ethidium staining. Follicles showed a decreased viability after freezing-thawing. After cryopreservation, a significant correlation between the percentage of normal follicles and viability rate was found using trypan blue (r = 0.82, p Conclusion We suggest the use of trypan blue staining for the histological assessment of viability, the use of LDH assay for the cytotoxicity assessement and finally the use of DNA fragmentation assessment to valid different freezing-thawing protocols.

  20. Viability and biomass of Micrococcus luteus DE2008 at different salinity concentrations determined by specific fluorochromes and CLSM-image analysis.

    Science.gov (United States)

    Puyen, Zully M; Villagrasa, Eduard; Maldonado, Juan; Esteve, Isabel; Solé, Antonio

    2012-01-01

    In previous studies, our group developed a method based on Confocal Laser Scanning Microscopy and Image Analysis (CLSM-IA) to analyze the diversity and biomass of cyanobacteria in microbial mats. However, this method cannot be applied to heterotrophic microorganisms, as these do not have autofluorescence. In this article, we present a method that combines CLSM-IA and Hoechst 33342 and SYTOX Green fluorochromes (FLU-CLSM-IA) to determine the viability and biomass of Micrococcus luteus DE2008, isolated from a saline microbial mat (Ebro Delta, Tarragona, Spain). The method has been applied to assess the effect of salinity on this microorganism. A reduction in viability and biomass (live cells) was observed as the salt concentration increases. The largest effect was at 100‰ NaCl with a cell death of 27.25% and a decrease in total and individual biomass of 39.75 and 0.009 mgC/cm(3), respectively, both with respect to optimal growth (10 ‰ NaCl). On the other hand, another important contribution of this article was that combining the FLU-CLSM-IA results with those achieved by plate counts enabled us to determine, for first time, the viability and the total biomass of the "dormant cells" (66.75% of viability and 40.59 mgC/cm(3) of total biomass at 100‰ NaCl). FLU-CLSM-IA is an efficient, fast, and reliable method for making a total count of cells at pixel level, including the dormant cells, to evaluate the viability and the biomass of a hetetrophic microorganism, M. luteus DE2008.

  1. A simple method to measure cell viability in proliferation and cytotoxicity assays

    Directory of Open Access Journals (Sweden)

    Ricardo Carneiro Borra

    2009-09-01

    Full Text Available Resazurin dye has been broadly used as indicator of cell viability in several types of assays for evaluation of the biocompatibility of medical and dental materials. Mitochondrial enzymes, as carriers of diaphorase activities, are probably responsible for the transference of electrons from NADPH + H+ to resazurin, which is reduced to resorufin. The level of reduction can be quantified by spectrophotometers since resazurin exhibits an absorption peak at 600 ηm and resorufin at 570 ηm wavelengths. However, the requirement of a spectrophotometer and specific filters for the quantification could be a barrier to many laboratories. Digital cameras containing red, green and blue filters, which allow the capture of red (600 to 700 ηm and green (500 to 600 ηm light wavelengths in ranges bordering on resazurin and resorufin absorption bands, could be used as an alternative method for the assessment of resazurin and resorufin concentrations. Thus, our aim was to develop a simple, cheap and precise method based on a digital CCD camera to measure the reduction of resazurin. We compared the capability of the CCD-based method to distinguish different concentrations of L929 and normal Human buccal fibroblast cell lines with that of a conventional microplate reader. The correlation was analyzed through the Pearson coefficient. The results showed a strong association between the measurements of the method developed here and those made with the microplate reader (r² = 0.996; p < 0.01 and with the cellular concentrations (r² = 0.965; p < 0.01. We concluded that the developed Colorimetric Quantification System based on CCD Images allowed rapid assessment of the cultured cell concentrations with simple equipment at a reduced cost.

  2. Determination of pollen viability, germination ratios and morphology ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-12-03

    Dec 3, 2008 ... Key words: Flower biology, malatya, Prunus armeniaca, scanning electron