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Sample records for high content cellular

  1. Quantitative high content imaging of cellular adaptive stress response pathways in toxicity for chemical safety assessment.

    Science.gov (United States)

    Wink, Steven; Hiemstra, Steven; Huppelschoten, Suzanna; Danen, Erik; Niemeijer, Marije; Hendriks, Giel; Vrieling, Harry; Herpers, Bram; van de Water, Bob

    2014-03-17

    Over the past decade, major leaps forward have been made on the mechanistic understanding and identification of adaptive stress response landscapes underlying toxic insult using transcriptomics approaches. However, for predictive purposes of adverse outcome several major limitations in these approaches exist. First, the limited number of samples that can be analyzed reduces the in depth analysis of concentration-time course relationships for toxic stress responses. Second these transcriptomics analysis have been based on the whole cell population, thereby inevitably preventing single cell analysis. Third, transcriptomics is based on the transcript level, totally ignoring (post)translational regulation. We believe these limitations are circumvented with the application of high content analysis of relevant toxicant-induced adaptive stress signaling pathways using bacterial artificial chromosome (BAC) green fluorescent protein (GFP) reporter cell-based assays. The goal is to establish a platform that incorporates all adaptive stress pathways that are relevant for toxicity, with a focus on drug-induced liver injury. In addition, cellular stress responses typically follow cell perturbations at the subcellular organelle level. Therefore, we complement our reporter line panel with reporters for specific organelle morphometry and function. Here, we review the approaches of high content imaging of cellular adaptive stress responses to chemicals and the application in the mechanistic understanding and prediction of chemical toxicity at a systems toxicology level.

  2. High content analysis at single cell level identifies different cellular responses dependent on nanomaterial concentrations.

    Science.gov (United States)

    Manshian, Bella B; Munck, Sebastian; Agostinis, Patrizia; Himmelreich, Uwe; Soenen, Stefaan J

    2015-09-08

    A mechanistic understanding of nanomaterial (NM) interaction with biological environments is pivotal for the safe transition from basic science to applied nanomedicine. NM exposure results in varying levels of internalized NM in different neighboring cells, due to variances in cell size, cell cycle phase and NM agglomeration. Using high-content analysis, we investigated the cytotoxic effects of fluorescent quantum dots on cultured cells, where all effects were correlated with the concentration of NMs at the single cell level. Upon binning the single cell data into different categories related to NM concentration, this study demonstrates, for the first time, that quantum dots activate both cytoprotective and cytotoxic mechanisms, resulting in a zero net result on the overall cell population, yet with significant effects in cells with higher cellular NM levels. Our results suggest that future NM cytotoxicity studies should correlate NM toxicity with cellular NM numbers on the single cell level, as conflicting mechanisms in particular cell subpopulations are commonly overlooked using classical toxicological methods.

  3. High content analysis at single cell level identifies different cellular responses dependent on nanomaterial concentrations

    Science.gov (United States)

    Manshian, Bella B.; Munck, Sebastian; Agostinis, Patrizia; Himmelreich, Uwe; Soenen, Stefaan J.

    2015-09-01

    A mechanistic understanding of nanomaterial (NM) interaction with biological environments is pivotal for the safe transition from basic science to applied nanomedicine. NM exposure results in varying levels of internalized NM in different neighboring cells, due to variances in cell size, cell cycle phase and NM agglomeration. Using high-content analysis, we investigated the cytotoxic effects of fluorescent quantum dots on cultured cells, where all effects were correlated with the concentration of NMs at the single cell level. Upon binning the single cell data into different categories related to NM concentration, this study demonstrates, for the first time, that quantum dots activate both cytoprotective and cytotoxic mechanisms, resulting in a zero net result on the overall cell population, yet with significant effects in cells with higher cellular NM levels. Our results suggest that future NM cytotoxicity studies should correlate NM toxicity with cellular NM numbers on the single cell level, as conflicting mechanisms in particular cell subpopulations are commonly overlooked using classical toxicological methods.

  4. Normalizing for individual cell population context in the analysis of high-content cellular screens

    Directory of Open Access Journals (Sweden)

    Knapp Bettina

    2011-12-01

    Full Text Available Abstract Background High-content, high-throughput RNA interference (RNAi offers unprecedented possibilities to elucidate gene function and involvement in biological processes. Microscopy based screening allows phenotypic observations at the level of individual cells. It was recently shown that a cell's population context significantly influences results. However, standard analysis methods for cellular screens do not currently take individual cell data into account unless this is important for the phenotype of interest, i.e. when studying cell morphology. Results We present a method that normalizes and statistically scores microscopy based RNAi screens, exploiting individual cell information of hundreds of cells per knockdown. Each cell's individual population context is employed in normalization. We present results on two infection screens for hepatitis C and dengue virus, both showing considerable effects on observed phenotypes due to population context. In addition, we show on a non-virus screen that these effects can be found also in RNAi data in the absence of any virus. Using our approach to normalize against these effects we achieve improved performance in comparison to an analysis without this normalization and hit scoring strategy. Furthermore, our approach results in the identification of considerably more significantly enriched pathways in hepatitis C virus replication than using a standard analysis approach. Conclusions Using a cell-based analysis and normalization for population context, we achieve improved sensitivity and specificity not only on a individual protein level, but especially also on a pathway level. This leads to the identification of new host dependency factors of the hepatitis C and dengue viruses and higher reproducibility of results.

  5. Cellular effect of high doses of silica-coated quantum dot profiled with high throughput gene expression analysis and high content cellomics measurements.

    Science.gov (United States)

    Zhang, Tingting; Stilwell, Jackie L; Gerion, Daniele; Ding, Lianghao; Elboudwarej, Omeed; Cooke, Patrick A; Gray, Joe W; Alivisatos, A Paul; Chen, Fanqing Frank

    2006-04-01

    Quantum dots (Qdots) are now used extensively for labeling in biomedical research, and this use is predicted to grow because of their many advantages over alternative labeling methods. Uncoated Qdots made of core/shell CdSe/ZnS are toxic to cells because of the release of Cd2+ ions into the cellular environment. This problem has been partially overcome by coating Qdots with polymers, poly(ethylene glycol) (PEG), or other inert molecules. The most promising coating to date, for reducing toxicity, appears to be PEG. When PEG-coated silanized Qdots (PEG-silane-Qdots) are used to treat cells, toxicity is not observed, even at dosages above 10-20 nM, a concentration inducing death when cells are treated with polymer or mercaptoacid coated Qdots. Because of the importance of Qdots in current and future biomedical and clinical applications, we believe it is essential to more completely understand and verify this negative global response from cells treated with PEG-silane-Qdots. Consequently, we examined the molecular and cellular response of cells treated with two different dosages of PEG-silane-Qdots. Human fibroblasts were exposed to 8 and 80 nM of these Qdots, and both phenotypic as well as whole genome expression measurements were made. PEG-silane-Qdots did not induce any statistically significant cell cycle changes and minimal apoptosis/necrosis in lung fibroblasts (IMR-90) as measured by high content image analysis, regardless of the treatment dosage. A slight increase in apoptosis/necrosis was observed in treated human skin fibroblasts (HSF-42) at both the low and the high dosages. We performed genome-wide expression array analysis of HSF-42 exposed to doses 8 and 80 nM to link the global cell response to a molecular and genetic phenotype. We used a gene array containing approximately 22,000 total probe sets, containing 18,400 probe sets from known genes. Only approximately 50 genes (approximately 0.2% of all the genes tested) exhibited a statistically significant

  6. Integrating high-content imaging and chemical genetics to probe host cellular pathways critical for Yersinia pestis infection.

    Directory of Open Access Journals (Sweden)

    Krishna P Kota

    Full Text Available The molecular machinery that regulates the entry and survival of Yersinia pestis in host macrophages is poorly understood. Here, we report the development of automated high-content imaging assays to quantitate the internalization of virulent Y. pestis CO92 by macrophages and the subsequent activation of host NF-κB. Implementation of these assays in a focused chemical screen identified kinase inhibitors that inhibited both of these processes. Rac-2-ethoxy-3 octadecanamido-1-propylphosphocholine (a protein Kinase C inhibitor, wortmannin (a PI3K inhibitor, and parthenolide (an IκB kinase inhibitor, inhibited pathogen-induced NF-κB activation and reduced bacterial entry and survival within macrophages. Parthenolide inhibited NF-κB activation in response to stimulation with Pam3CSK4 (a TLR2 agonist, E. coli LPS (a TLR4 agonist or Y. pestis infection, while the PI3K and PKC inhibitors were selective only for Y. pestis infection. Together, our results suggest that phagocytosis is the major stimulus for NF-κB activation in response to Y. pestis infection, and that Y. pestis entry into macrophages may involve the participation of protein kinases such as PI3K and PKC. More importantly, the automated image-based screening platform described here can be applied to the study of other bacteria in general and, in combination with chemical genetic screening, can be used to identify host cell functions facilitating the identification of novel antibacterial therapeutics.

  7. Cellular Effect of High Doses of Silica-Coated Quantum Dot Profiled with High Throughput Gene Expression Analysis and High Content Cellomics Measurements

    OpenAIRE

    Zhang, Tingting; Stilwell, Jackie L.; Gerion, Daniele; Ding, Lianghao; Elboudwarej, Omeed; Cooke, Patrick A.; Gray, Joe W.; Alivisatos, A. Paul; Chen, Fanqing Frank

    2006-01-01

    Quantum dots (Qdots) are now used extensively for labeling in biomedical research, and this use is predicted to grow because of their many advantages over alternative labeling methods. Uncoated Qdots made of core/shell CdSe/ZnS are toxic to cells because of the release of Cd2+ ions into the cellular environment. This problem has been partially overcome by coating Qdots with polymers, poly(ethylene glycol) (PEG), or other inert molecules. The most promising coating to date, for reducing toxici...

  8. Cellular content and biosynthesis of polyamines during rooster spermatogenesis.

    Science.gov (United States)

    Oliva, R; Vidal, S; Mezquita, C

    1982-11-15

    The natural polyamines spermine and spermidine, and the diamine putrescine, were extracted from rooster testis cells separated by sedimentation at unit gravity, and from vas-deferens spermatozoa. The ratios spermine/DNA and spermidine/DNA were kept relatively constant throughout spermatogenesis, whereas the ratio putrescine/DNA rose in elongated spermatids. The cellular content of spermine, spermidine and putrescine decreased markedly in mature spermatozoa. Two rate-limiting enzymes in the biosynthetic pathway of polyamines, ornithine decarboxylase and S-adenosyl-L-methionine decarboxylase, showed their highest activities at the end of spermiogenesis and were not detectable in vas-deferens spermatozoa. A marked reduction in cell volume during spermiogenesis without a parallel decrease in the cellular content of polyamines suggests the possibility that the marked changes in chromatin composition and structure occurring in rooster late spermatids could take place in an ambience of high polyamine concentration.

  9. Cellular content of biomolecules in sub-seafloor microbial communities

    Science.gov (United States)

    Braun, Stefan; Morono, Yuki; Becker, Kevin W.; Hinrichs, Kai-Uwe; Kjeldsen, Kasper U.; Jørgensen, Bo B.; Lomstein, Bente Aa.

    2016-09-01

    Microbial biomolecules, typically from the cell envelope, can provide crucial information about distribution, activity, and adaptations of sub-seafloor microbial communities. However, when cells die these molecules can be preserved in the sediment on timescales that are likely longer than the lifetime of their microbial sources. Here we provide for the first time measurements of the cellular content of biomolecules in sedimentary microbial cells. We separated intact cells from sediment matrices in samples from surficial, deeply buried, organic-rich, and organic-lean marine sediments by density centrifugation. Amino acids, amino sugars, muramic acid, and intact polar lipids were analyzed in both whole sediment and cell extract, and cell separation was optimized and evaluated in terms of purity, separation efficiency, taxonomic resemblance, and compatibility to high-performance liquid chromatography and mass spectrometry for biomolecule analyses. Because cell extracts from density centrifugation still contained considerable amounts of detrital particles and non-cellular biomolecules, we further purified cells from two samples by fluorescence-activated cell sorting (FACS). Cells from these highly purified cell extracts had an average content of amino acids and lipids of 23-28 fg cell-1 and 2.3 fg cell-1, respectively, with an estimated carbon content of 19-24 fg cell-1. In the sediment, the amount of biomolecules associated with vegetative cells was up to 70-fold lower than the total biomolecule content. We find that the cellular content of biomolecules in the marine subsurface is up to four times lower than previous estimates. Our approach will facilitate and improve the use of biomolecules as proxies for microbial abundance in environmental samples and ultimately provide better global estimates of microbial biomass.

  10. Cellular content of biomolecules in sub-seafloor microbial communities

    DEFF Research Database (Denmark)

    Braun, Stefan; Morono, Yuki; Becker, Kevin W.;

    2016-01-01

    Microbial biomolecules, typically from the cell envelope, can provide crucial information about distribution, activity, and adaptations of sub-seafloor microbial communities. However, when cells die these molecules can be preserved in the sediment on timescales that are likely longer than...... the lifetime of their microbial sources. Here we provide for the first time measurements of the cellular content of biomolecules in sedimentary microbial cells. We separated intact cells from sediment matrices in samples from surficial, deeply buried, organic-rich, and organic-lean marine sediments by density...... and mass spectrometry for biomolecule analyses. Because cell extracts from density centrifugation still contained considerable amounts of detrital particles and non-cellular biomolecules, we further purified cells from two samples by fluorescence-activated cell sorting (FACS). Cells from these highly...

  11. Regulation and quantification of cellular mitochondrial morphology and content.

    Science.gov (United States)

    Tronstad, Karl J; Nooteboom, Marco; Nilsson, Linn I H; Nikolaisen, Julie; Sokolewicz, Maciek; Grefte, Sander; Pettersen, Ina K N; Dyrstad, Sissel; Hoel, Fredrik; Willems, Peter H G M; Koopman, Werner J H

    2014-01-01

    Mitochondria play a key role in signal transduction, redox homeostasis and cell survival, which extends far beyond their classical functioning in ATP production and energy metabolism. In living cells, mitochondrial content ("mitochondrial mass") depends on the cell-controlled balance between mitochondrial biogenesis and degradation. These processes are intricately linked to changes in net mitochondrial morphology and spatiotemporal positioning ("mitochondrial dynamics"), which are governed by mitochondrial fusion, fission and motility. It is becoming increasingly clear that mitochondrial mass and dynamics, as well as its ultrastructure and volume, are mechanistically linked to mitochondrial function and the cell. This means that proper quantification of mitochondrial morphology and content is of prime importance in understanding mitochondrial and cellular physiology in health and disease. This review first presents how cellular mitochondrial content is regulated at the level of mitochondrial biogenesis, degradation and dynamics. Next we discuss how mitochondrial dynamics and content can be analyzed with a special emphasis on quantitative live-cell microscopy strategies.

  12. Effects of defined mixtures of persistent organic pollutants (POPs) on multiple cellular responses in the human hepatocarcinoma cell line, HepG2, using high content analysis screening.

    Science.gov (United States)

    Wilson, Jodie; Berntsen, Hanne Friis; Zimmer, Karin Elisabeth; Frizzell, Caroline; Verhaegen, Steven; Ropstad, Erik; Connolly, Lisa

    2016-03-01

    Persistent organic pollutants (POPs) are toxic substances, highly resistant to environmental degradation, which can bio-accumulate and have long-range atmospheric transport potential. Most studies focus on single compound effects, however as humans are exposed to several POPs simultaneously, investigating exposure effects of real life POP mixtures on human health is necessary. A defined mixture of POPs was used, where the compound concentration reflected its contribution to the levels seen in Scandinavian human serum (total mix). Several sub mixtures representing different classes of POPs were also constructed. The perfluorinated (PFC) mixture contained six perfluorinated compounds, brominated (Br) mixture contained seven brominated compounds, chlorinated (Cl) mixture contained polychlorinated biphenyls and also p,p'-dichlorodiphenyldichloroethylene, hexachlorobenzene, three chlordanes, three hexachlorocyclohexanes and dieldrin. Human hepatocarcinoma (HepG2) cells were used for 2h and 48h exposures to the seven mixtures and analysis on a CellInsight™ NXT High Content Screening platform. Multiple cytotoxic endpoints were investigated: cell number, nuclear intensity and area, mitochondrial mass and membrane potential (MMP) and reactive oxygen species (ROS). Both the Br and Cl mixtures induced ROS production but did not lead to apoptosis. The PFC mixture induced ROS production and likely induced cell apoptosis accompanied by the dissipation of MMP. Synergistic effects were evident for ROS induction when cells were exposed to the PFC+Br mixture in comparison to the effects of the individual mixtures. No significant effects were detected in the Br+Cl, PFC+Cl or total mixtures, which contain the same concentrations of chlorinated compounds as the Cl mixture plus additional compounds; highlighting the need for further exploration of POP mixtures in risk assessment. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Hierarchical Cellular Structures in High-Capacity Cellular Communication Systems

    CERN Document Server

    Jain, R K; Agrawal, N K

    2011-01-01

    In the prevailing cellular environment, it is important to provide the resources for the fluctuating traffic demand exactly in the place and at the time where and when they are needed. In this paper, we explored the ability of hierarchical cellular structures with inter layer reuse to increase the capacity of mobile communication network by applying total frequency hopping (T-FH) and adaptive frequency allocation (AFA) as a strategy to reuse the macro and micro cell resources without frequency planning in indoor pico cells [11]. The practical aspects for designing macro- micro cellular overlays in the existing big urban areas are also explained [4]. Femto cells are inducted in macro / micro / pico cells hierarchical structure to achieve the required QoS cost effectively.

  14. Relationship between histologic grade and cytofluorometric cellular DNA and RNA content in primary bone tumors.

    Science.gov (United States)

    Takeshita, H; Kusuzaki, K; Kuzuhara, A; Tsuji, Y; Ashihara, T; Gebhardt, M C; Mankin, H J; Springfield, D S; Hirasawa, Y

    2001-01-01

    The diagnosis and grading of bone tumors remains a challenging problem. We studied the relationship between histologic grade and cytofluorometric cellular DNA and RNA content in 108 primary bone tumors. The data included DNA ploidy, mean DNA content (MDC), S-phase fraction (SPF), mean RNA content (MRC) and RNA/DNA ratio (RDR; MRC/MDC) which represents the RNA content normalized for the DNA content. Benign tumors had a diploid stem line with low MDC (mean; 1.04), low SPF (0.9), high MRC (2.41) and high RDR (2.31). Giant cell tumors of bone, which are locally aggressive benign tumors, showed diploidy with relatively higher MDC (1.07, p osteosarcomas revealed aneuploidy with remarkably higher MDC (1.70 in osteosarcomas, p < 0.01) and SPF (6.5, p < 0.01), but lower RDR (1.70, p < 0.01). In contrast, small cell sarcomas, such as Ewing's sarcomas, showed diploidy with low MDC (1.11 in Ewing's sarcomas, N.S.) and SPF (2.5, p < 0.01) and extremely low RDR (1.34, p < 0.01). The RDR value was higher in well-differentiated tumors than in primitive tumors, rendering it useful in grading bone tumors with a diploid stem line. By combining the RDR value with the MDC value, 96% of diploid sarcomas could be distinguished from benign tumors. These results indicate that cellular DNA and RNA content analysis may be of value in assessing the malignant potential of diploid as well as aneuploid bone sarcomas.

  15. Regulation and quantification of cellular mitochondrial morphology and content

    NARCIS (Netherlands)

    Tronstad, K.J.; Nooteboom, M.; Nilsson, L.I.; Nikolaisen, J.; Sokolewicz, M.; Grefte, S.; Pettersen, I.K.; Dyrstad, S.; Hoel, F.; Willems, P.H.G.M.; Koopman, W.J.H.

    2014-01-01

    Mitochondria play a key role in signal transduction, redox homeostasis and cell survival, which extends far beyond their classical functioning in ATP production and energy metabolism. In living cells, mitochondrial content ("mitochondrial mass") depends on the cell-controlled balance between mitocho

  16. Regulation and quantification of cellular mitochondrial morphology and content

    NARCIS (Netherlands)

    Tronstad, K.J.; Nooteboom, M.; Nilsson, L.I.; Nikolaisen, J.; Sokolewicz, M.; Grefte, S.; Pettersen, I.K.; Dyrstad, S.; Hoel, F.; Willems, P.H.G.M.; Koopman, W.J.H.

    2014-01-01

    Mitochondria play a key role in signal transduction, redox homeostasis and cell survival, which extends far beyond their classical functioning in ATP production and energy metabolism. In living cells, mitochondrial content ("mitochondrial mass") depends on the cell-controlled balance between

  17. Influence of Bottom Ash Replacements as Fine Aggregate on the Property of Cellular Concrete with Various Foam Contents

    Directory of Open Access Journals (Sweden)

    Patchara Onprom

    2015-01-01

    Full Text Available This research focuses on evaluating the feasibility of utilizing bottom ash from coal burning power plants as a fine aggregate in cellular concrete with various foam contents. Flows of all mixtures were controlled within 45 ± 5% and used foam content at 30%, 40%, 50%, 60%, and 70% by volume of mixture. Bottom ash from Mae Moh power plant in Thailand was used to replace river sand at the rates of 0%, 25%, 50%, 75%, and 100% by volume of sand. Compressive strength, water absorption, and density of cellular concretes were determined at the ages of 7, 14, and 28 days. Nonlinear regression technique was developed to construct the mathematical models for predicting the compressive strength, water absorption, and density of cellular concrete. The results revealed that the density of cellular concrete decreased while the water absorption increased with an increase in replacement level of bottom ash. From the experimental results, it can be concluded that bottom ash can be used as fine aggregate in the cellular concrete. In addition, the nonlinear regression models give very high degree of accuracy (R2>0.99.

  18. Shedding light on filovirus infection with high-content imaging.

    Science.gov (United States)

    Pegoraro, Gianluca; Bavari, Sina; Panchal, Rekha G

    2012-08-01

    Microscopy has been instrumental in the discovery and characterization of microorganisms. Major advances in high-throughput fluorescence microscopy and automated, high-content image analysis tools are paving the way to the systematic and quantitative study of the molecular properties of cellular systems, both at the population and at the single-cell level. High-Content Imaging (HCI) has been used to characterize host-virus interactions in genome-wide reverse genetic screens and to identify novel cellular factors implicated in the binding, entry, replication and egress of several pathogenic viruses. Here we present an overview of the most significant applications of HCI in the context of the cell biology of filovirus infection. HCI assays have been recently implemented to quantitatively study filoviruses in cell culture, employing either infectious viruses in a BSL-4 environment or surrogate genetic systems in a BSL-2 environment. These assays are becoming instrumental for small molecule and siRNA screens aimed at the discovery of both cellular therapeutic targets and of compounds with anti-viral properties. We discuss the current practical constraints limiting the implementation of high-throughput biology in a BSL-4 environment, and propose possible solutions to safely perform high-content, high-throughput filovirus infection assays. Finally, we discuss possible novel applications of HCI in the context of filovirus research with particular emphasis on the identification of possible cellular biomarkers of virus infection.

  19. Shedding Light on Filovirus Infection with High-Content Imaging

    Directory of Open Access Journals (Sweden)

    Rekha G. Panchal

    2012-08-01

    Full Text Available Microscopy has been instrumental in the discovery and characterization of microorganisms. Major advances in high-throughput fluorescence microscopy and automated, high-content image analysis tools are paving the way to the systematic and quantitative study of the molecular properties of cellular systems, both at the population and at the single-cell level. High-Content Imaging (HCI has been used to characterize host-virus interactions in genome-wide reverse genetic screens and to identify novel cellular factors implicated in the binding, entry, replication and egress of several pathogenic viruses. Here we present an overview of the most significant applications of HCI in the context of the cell biology of filovirus infection. HCI assays have been recently implemented to quantitatively study filoviruses in cell culture, employing either infectious viruses in a BSL-4 environment or surrogate genetic systems in a BSL-2 environment. These assays are becoming instrumental for small molecule and siRNA screens aimed at the discovery of both cellular therapeutic targets and of compounds with anti-viral properties. We discuss the current practical constraints limiting the implementation of high-throughput biology in a BSL-4 environment, and propose possible solutions to safely perform high-content, high-throughput filovirus infection assays. Finally, we discuss possible novel applications of HCI in the context of filovirus research with particular emphasis on the identification of possible cellular biomarkers of virus infection.

  20. Cellular content of ubiquitin and formation of ubiquitin conjugates during chicken spermatogenesis.

    Science.gov (United States)

    Agell, N; Mezquita, C

    1988-03-15

    Ubiquitin was purified from chicken testis and its content, biosynthesis and formation of conjugates was determined in germinal cells at successive stages of spermatogenesis. Free ubiquitin increased markedly during spermatogenesis, reaching its maximum level in early spermatids. High levels of ubiquitin were still present in late spermatids but were not detectable in mature spermatozoa. Biosynthesis of ubiquitin occurred in vitro in a fraction containing meiotic and pre-meiotic cells, and during spermiogenesis, in early and late spermatids. The cellular content of free ubiquitin increased after ATP depletion, especially in early spermatids. Lysates of chicken testis cells, particularly those obtained from spermatids, were able to form nuclear (24 and 27 kDa) and extranuclear (55-90 kDa) ubiquitin conjugates in vitro. The presence of increasing levels of ubiquitin and ubiquitin conjugates in chicken spermatids may suggest a possible involvement of this protein in the marked changes of protein turnover, chromatin structure and cell-cell interactions that spermatids undergo during spermiogenesis.

  1. Effect of cellular inositol content on ethanol tolerance of Saccharomyces cerevisiae in sake brewing.

    Science.gov (United States)

    Furukawa, Keiji; Kitano, Hideyuki; Mizoguchi, Haruhiko; Hara, Shodo

    2004-01-01

    The effect of cellular inositol content on the ethanol tolerance of sake yeast was investigated. In a static culture of strain K901 in a synthetic medium, when cells were grown in the presence of inositol in limited amount (L-cells), the inositol content of cells decreased by one-third that of cells grown in the presence of inositol in sufficient amount (H-cells). L-cells exhibited a higher death rate constant than H-cells in the presence of 12-20% ethanol, while no difference in specific ethanol production rate in the presence of 0-18% ethanol between the two cell types was observed. L-cells leaked more intracellular components, such as nucleotides, phosphate and potassium, in the presence of ethanol than H-cells. L-cells exhibited a lower intracellular pH value than H-cells, which represented the lowering of cell vitality by the decrease in H(+) extrusion activity. Furthermore, the plasma membrane H(+)-ATPase activity of L-cells was approximately one-half of that of H-cells. Therefore, it was considered that the decrease in viability in the presence of ethanol due to inositol limitation results from the lowering of H(+)-ATPase activity, which maintains the permeability barrier of the yeast membrane, ensuring the homeostasis of ions in the cytoplasm of yeast cells. It is assumed that the lowering of H(+)-ATPase activity due to inositol limitation is caused by the change in lipid environment of the enzyme, which is affected by inositol-containing glycerophospholipids such as phosphatidylinositol (PI), because in the PI-saturated mixed micellar assay system, the difference in H(+)-ATPase activity between L- and H-cells disappeared. In the early stage of sake mash, inositol limitation lowers the ethanol tolerance due to the decrease in H(+)-ATPase activity as in static culture. In the final stage of sake mash, the disruption of the ino1 gene responsible for inositol synthesis, resulted in a decrease in cell density. Furthermore, the ino1 disruptant, which was not

  2. Modulation of the cellular content of metabolites in adipocytes by insulin.

    Science.gov (United States)

    Qiao, Yuhang; Tomonaga, Shozo; Matsui, Tohru; Funaba, Masayuki

    2016-03-15

    Although the insulin-mediated cell signaling pathway has been extensively examined, changes in the cellular content of metabolites currently remain unclear. We herein examined metabolite contents in 3T3-L1 adipocytes treated with insulin using a metabolomic analysis. Fifty-four compounds were detected, and the contents of metabolites from the citric acid cycle increased in response to the insulin treatment for 4 h, which was sensitive to U0126 and LY294002, inhibitors for mitogen-activated protein kinase kinase-1 and phosphoinositide 3-kinase, respectively. The cellular contents of fumaric acid and malic acid were increased more by insulin than those of citric acid and succinic acid. Time-course changes in metabolites from the citric acid cycle exhibited oscillations with a 2-h cycle. A metabolic pathway analysis also indicated that insulin affected the metabolism of alanine, aspartate and glutamate, as well as that of arginine and proline. The contents of free amino acids were slightly decreased by the insulin treatment, while the co-treatment with U0126 and LY294002 abrogated these insulin-mediated decreases. The present study revealed the unexpected accumulation of citric acid cycle metabolites in adipocytes by insulin. Our results indicate the usefulness of metabolomic analyses for obtaining a more comprehensive understanding of the regulation of metabolic pathways in cell-culture systems. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  3. Development of automatic image analysis methods for high-throughput and high-content screening

    NARCIS (Netherlands)

    Di, Zi

    2013-01-01

    This thesis focuses on the development of image analysis methods for ultra-high content analysis of high-throughput screens where cellular phenotype responses to various genetic or chemical perturbations that are under investigation. Our primary goal is to deliver efficient and robust image analysis

  4. Development of automatic image analysis methods for high-throughput and high-content screening

    NARCIS (Netherlands)

    Di, Zi

    2013-01-01

    This thesis focuses on the development of image analysis methods for ultra-high content analysis of high-throughput screens where cellular phenotype responses to various genetic or chemical perturbations that are under investigation. Our primary goal is to deliver efficient and robust image analysis

  5. Cellular uptake and cytotoxic potential of respirable bentonite particles with different quartz contents and chemical modifications in human lung fibroblasts

    Energy Technology Data Exchange (ETDEWEB)

    Geh, Stefan; Rettenmeier, Albert W.; Dopp, Elke [University Hospital, Institute of Hygiene and Occupational Medicine, Essen (Germany); Yuecel, Raif [University Hospital, Institute of Cell Biology (Cancer Research), Essen (Germany); Duffin, Rodger [Institute of Environmental Health Research (IUF), Duesseldorf (Germany); University of Edinburgh, ELEGI COLT Lab, Scotland (United Kingdom); Albrecht, Catrin; Borm, Paul J.A. [Institute of Environmental Health Research (IUF), Duesseldorf (Germany); Armbruster, Lorenz [Verein fuer Technische Sicherheit und Umweltschutz e.V., Gotha (Germany); Raulf-Heimsoth, Monika; Bruening, Thomas [Research Institute for Occupational Medicine of the Institutions for Statutory Accident Insurance and Prevention (BGFA), Bochum (Germany); Hoffmann, Eik [University of Rostock, Institute of Biology, Department of Cell Biology and Biosystems Technology, Rostock (Germany)

    2006-02-01

    Considering the biological reactivity of pure quartz in lung cells, there is a strong interest to clarify the cellular effects of respirable siliceous dusts, like bentonites. In the present study, we investigated the cellular uptake and the cytotoxic potential of bentonite particles (Oe< 10 {mu}m) with an {alpha}-quartz content of up to 6% and different chemical modifications (activation: alkaline, acidic, organic) in human lung fibroblasts (IMR90). Additionally, the ability of the particles to induce apoptosis in IMR90-cells and the hemolytic activity was tested. All bentonite samples were tested for endotoxins with the in vitro-Pyrogen test and were found to be negative. Cellular uptake of particles by IMR90-cells was studied by transmission electron microscopy (TEM). Cytotoxicity was analyzed in IMR90-cells by determination of viable cells using flow cytometry and by measuring of the cell respiratory activity. Induced apoptotic cells were detected by AnnexinV/Propidiumiodide-staining and gel electrophoresis. Our results demonstrate that activated bentonite particles are better taken up by IMR90-cells than untreated (native) bentonite particles. Also, activated bentonite particles with a quartz content of 5-6% were more cytotoxic than untreated bentonites or bentonites with a quartz content lower than 4%. The bentonite samples induced necrotic as well as apoptotic cell death. In general, bentonites showed a high membrane-damaging potential shown as hemolytic activity in human erythrocytes. We conclude that cellular effects of bentonite particles in human lung cells are enhanced after chemical treatment of the particles. The cytotoxic potential of the different bentonites is primarily characterized by a strong lysis of the cell membrane. (orig.)

  6. A high-efficiency cellular extraction system for biological proteomics

    OpenAIRE

    2015-01-01

    Recent developments in quantitative high-resolution mass spectrometry have led to significant improvements in the sensitivity and specificity of biochemical analyses of cellular reactions, protein-protein interactions, and small molecule drug discovery. These approaches depend on cellular proteome extraction that preserves native protein activities. Here, we systematically analyzed mechanical methods of cell lysis and physical protein extraction to identify those that maximize the extraction ...

  7. Polyamines regulate cell growth and cellular methylglyoxal in high-glucose medium independently of intracellular glutathione.

    Science.gov (United States)

    Kwak, Min-Kyu; Lee, Mun-Hyoung; Park, Seong-Jun; Shin, Sang-Min; Liu, Rui; Kang, Sa-Ouk

    2016-03-01

    Polyamines can presumably inhibit protein glycation, when associated with the methylglyoxal inevitably produced during glycolysis. Herein, we hypothesized a nonenzymatic interaction between putrescine and methylglyoxal in putrescine-deficient or -overexpressing Dictyostelium cells in high-glucose medium, which can control methylglyoxal production. Putrescine was essentially required for growth rescue accompanying methylglyoxal detoxification when cells underwent growth defect and cell cycle G1-arrest when supplemented with high glucose. Furthermore, methylglyoxal regulation by putrescine seemed to be a parallel pathway independent of the changes in cellular glutathione content in high-glucose medium. Consequently, we suggest that Dictyostelium cells need polyamines for normal growth and cellular methylglyoxal regulation.

  8. Analysis of Cellular DNA Content by Flow and Laser Scanning Cytometry

    Science.gov (United States)

    Darzynkiewicz, Zbigniew; Halicka, H. Dorota; Zhao, Hong

    2010-01-01

    This chapter covers several aspects of methodology of DNA content analysis in individual cells that is most commonly used for assessment of DNA ploidy and for enumeration of cells in particular phases of the cell cycle. Briefly presented are general principles of instrumentation and cell analysis by flow- and laser scanning- cytometry. Described are major methods designed to stain DNA with fluorochromes in live cells, in detergent-permeabilized cells, in cells fixed prior to DNA staining as well as in nuclei of cells isolated from paraffin-embedded tissues. Briefly addressed are approaches to estimate cellular DNA content in conjunction with cellular immunophenotype. Discussed are factors that affect accuracy of DNA content measurement such as: (i) differences in chromatin structure of the analyzed cells that restrict DNA accessibility to fluorochromes, (ii) stoichiometry of interaction between fluorochromes and DNA in chromatin and (iii) chemical mass action law defining dependency of fluorochrome binding to DNA in relation to fluorochrome concentration and number of potential binding sites in a sample. Described also are controls used to ensure accuracy of DNA ploidy determination, the principles in ploidy assessment and possible pitfalls in analysis. PMID:20687474

  9. High content analysis in amyotrophic lateral sclerosis.

    Science.gov (United States)

    Rinaldi, Federica; Motti, Dario; Ferraiuolo, Laura; Kaspar, Brian K

    2017-04-01

    Amyotrophic lateral sclerosis (ALS) is a devastating disease characterized by the progressive loss of motor neurons. Neurons, astrocytes, oligodendrocytes and microglial cells all undergo pathological modifications in the onset and progression of ALS. A number of genes involved in the etiopathology of the disease have been identified, but a complete understanding of the molecular mechanisms of ALS has yet to be determined. Currently, people affected by ALS have a life expectancy of only two to five years from diagnosis. The search for a treatment has been slow and mostly unsuccessful, leaving patients in desperate need of better therapies. Until recently, most pre-clinical studies utilized the available ALS animal models. In the past years, the development of new protocols for isolation of patient cells and differentiation into relevant cell types has provided new tools to model ALS, potentially more relevant to the disease itself as they directly come from patients. The use of stem cells is showing promise to facilitate ALS research by expanding our understanding of the disease and help to identify potential new therapeutic targets and therapies to help patients. Advancements in high content analysis (HCA) have the power to contribute to move ALS research forward by combining automated image acquisition along with digital image analysis. With modern HCA machines it is possible, in a period of just a few hours, to observe changes in morphology and survival of cells, under the stimulation of hundreds, if not thousands of drugs and compounds. In this article, we will summarize the major molecular and cellular hallmarks of ALS, describe the advancements provided by the in vitro models developed in the last few years, and review the studies that have applied HCA to the ALS field to date. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Growth and cellular ion content of a salt-sensitive symbiotic system Azolla pinnata-Anabaena azollae under NaCl stress.

    Science.gov (United States)

    Rai, Vandna; Sharma, Naveen Kumar; Rai, Ashwani K

    2006-09-01

    Salinity, at a concentration of 10 mM NaCl affected the growth of Azolla pinnata-Anabaena azollae association and became lethal at 40 mM. Plants exposed up to 30 mM NaCl exhibited longer roots than the control, especially during the beginning of incubation. Average root number in plants exposed to 10 and 20 mM NaCl remained almost the same as in control. A further rise in NaCl concentration to 30 mM reduced the root number, and roots shed off at 40 mM NaCl. Presence of NaCl in the nutrient solution increased the cellular Na+ of the intact association exhibiting differential accumulation by individual partners, while it reduced the cellular Ca2+ level. However, cellular K+ content did not show significant change. Cellular Na+ based on fresh weight of respective individual partners (host tissues and cyanobiont) remained higher in the host tissues than the cyanobiont, while reverse was true for K+ and Ca2+ contents. The contribution of A. azollae in the total cellular ion content of the association was a little because of meagre contribution of the cyanobiont mass (19-21%). High salt sensitivity of Azolla-Anabaena complex is due to an inability of the association to maintain low Na+ and high Ca2+ cellular level.

  11. Immunochemical determination of cellular content of translation release factor RF4 in Escherichia coli

    DEFF Research Database (Denmark)

    Andersen, Lars Dyrskjøt; Manuel Palacios Moreno, Juan; Clark, Brian F. C.

    1999-01-01

    of the stop codons, and RF3 is known to accelerate the overall termination process. Release factor RF4 is a protein involved in the release of the mRNA and tRNA from the ribosomal complex. Furthermore, RF4 is involved in the proofreading in the elongation step of protein biosynthesis. The cellular contents...... of RF1, RF2, and RF3 were determined earlier. Here we report the cellular content of RF4 in Escherichia coli to be approximately 16,500 molecules per cell. The cells were grown in a rich medium and harvested in the beginning of the exponential growth phase. The quantifications were performed by using......The biosynthesis of proteins in prokaryotes is terminated when a stop codon is present in the A-site of the 70S ribosomal complex. Four different translation termination factors are known to participate in the termination process. Release factor RF1 and RF2 are responsible for the recognition...

  12. High-throughput screening for modulators of cellular contractile force

    CERN Document Server

    Park, Chan Young; Tambe, Dhananjay; Chen, Bohao; Lavoie, Tera; Dowell, Maria; Simeonov, Anton; Maloney, David J; Marinkovic, Aleksandar; Tschumperlin, Daniel J; Burger, Stephanie; Frykenberg, Matthew; Butler, James P; Stamer, W Daniel; Johnson, Mark; Solway, Julian; Fredberg, Jeffrey J; Krishnan, Ramaswamy

    2014-01-01

    When cellular contractile forces are central to pathophysiology, these forces comprise a logical target of therapy. Nevertheless, existing high-throughput screens are limited to upstream signaling intermediates with poorly defined relationship to such a physiological endpoint. Using cellular force as the target, here we screened libraries to identify novel drug candidates in the case of human airway smooth muscle cells in the context of asthma, and also in the case of Schlemm's canal endothelial cells in the context of glaucoma. This approach identified several drug candidates for both asthma and glaucoma. We attained rates of 1000 compounds per screening day, thus establishing a force-based cellular platform for high-throughput drug discovery.

  13. A High-Efficiency Cellular Extraction System for Biological Proteomics.

    Science.gov (United States)

    Dhabaria, Avantika; Cifani, Paolo; Reed, Casie; Steen, Hanno; Kentsis, Alex

    2015-08-07

    Recent developments in quantitative high-resolution mass spectrometry have led to significant improvements in the sensitivity and specificity of the biochemical analyses of cellular reactions, protein-protein interactions, and small-molecule-drug discovery. These approaches depend on cellular proteome extraction that preserves native protein activities. Here, we systematically analyzed mechanical methods of cell lysis and physical protein extraction to identify those that maximize the extraction of cellular proteins while minimizing their denaturation. Cells were mechanically disrupted using Potter-Elvehjem homogenization, probe- or adaptive-focused acoustic sonication, and were in the presence of various detergents, including polyoxyethylene ethers and esters, glycosides, and zwitterions. Using fluorescence spectroscopy, biochemical assays, and mass spectrometry proteomics, we identified the combination of adaptive focused acoustic (AFA) sonication in the presence of a binary poloxamer-based mixture of octyl-β-glucoside and Pluronic F-127 to maximize the depth and yield of the proteome extraction while maintaining native protein activity. This binary poloxamer extraction system allowed for native proteome extraction comparable in coverage to the proteomes extracted using denaturing SDS or guanidine-containing buffers, including the efficient extraction of all major cellular organelles. This high-efficiency cellular extraction system should prove useful for a variety of cell biochemical studies, including structural and functional proteomics.

  14. High-throughput microcavitation bubble induced cellular mechanotransduction

    Science.gov (United States)

    Compton, Jonathan Lee

    Focused pulsed laser irradiation allows for the deposition of energy with high spatial and temporal resolution. These attributes provide an optimal tool for non-contact manipulation in cellular biology such as laser microsurgery, cell membrane permeabilization, as well as targeted cell death. In this thesis we investigate the direct physical effects produced by laser- generated microcavitation bubbles in adherent cell cultures. We examine how variation in pulse durations (180 ps - 6ns) and pulse energy (0.5 - 40 mu;J) affect microcavitation bubble (mu;CB) generated cell lysis, necrosis, and molecular delivery. To compare the effects of pulse duration we employ classical fluid dynamics modeling to quantify the perturbation caused on cell populations from mu;CB generated microTsunamis (a transient microscale burst of hydrodynamic shear stress). Through time-resolved imaging we capture the mu;CB dynamics at various energies and pulse durations. Moreover, the mathematical modeling provides information regarding the cellular exposure to time varying shear stress and impulse as a function of radial location from the mu;CB center. We demonstrate that the resultant cellular effect can be predicted based on the total impulse across a two order of magnitude span of pulse duration and pulse energy. We also examine the region of cells beyond the zone of molecular delivery to investigate possible cellular reactions to mu;Tsunami exposure. Our studies have shown that cellular mechanotransduction occurs within cell populations spanning an area of up to 1 mm2 surrounding the mu;CB. Visualization of mechanotransduction is achieved through the visualization of intracellular calcium signaling via fluorescence microscopy that occurs due to the ability of the muTsunami generated shear stresses to stimulate G-protein coupled receptors at the apical cell surface. Moreover, we have shown that the observed signaling can be attenuated in a dose-dependent manner using 2-APB which is a known

  15. A novel cellular automata based technique for visual multimedia content encryption

    Science.gov (United States)

    Chatzichristofis, Savvas A.; Mitzias, Dimitris A.; Sirakoulis, Georgios Ch.; Boutalis, Yiannis S.

    2010-11-01

    This paper proposes a new method for visual multimedia content encryption using Cellular Automata (CA). The encryption scheme is based on the application of an attribute of the CLF XOR filter, according to which the original content of a cellular neighborhood can be reconstructed following a predetermined number of repeated applications of the filter. The encryption is achieved using a key image of the same dimensions as the image being encrypted. This technique is accompanied by the one-time pad (OTP) encryption method, rendering the proposed method reasonably powerful, given the very large number of resultant potential security keys. The method presented here makes encryption possible in cases where there is more than one image with the use of just one key image. A further significant characteristic of the proposed method is that it demonstrates how techniques from the field of image retrieval can be used in the field of image encryption. The proposed method is further strengthened by the fact that the resulting encrypted image for a given key image is different each time. The encryption result depends on the structure of an artificial image produced by the superposition of four 1-D CA time-space diagrams as well as from a CA random number generator. A semi-blind source separation algorithm is used to decrypt the encrypted image. The result of the decryption is a lossless representation of the encrypted image. Simulation results demonstrate the effectiveness of the proposed encryption method. The proposed method is implemented in C# and is available online through the img(Rummager) application.

  16. COPPER STRESS ON CELLULAR CONTENTS AND FATTY ACID PROFILES IN CHLORELLA SPECIES

    Directory of Open Access Journals (Sweden)

    G. Sibi

    2014-01-01

    Full Text Available Higher photosynthetic efficiency and biomass production with rapid growth makes microalgae as potential candidates over other energy crops in many applications. Heavy metals influence the production of secondary metabolites and lipd content of microalgae in particular. A study was conducted using six Chlorella species under heavy metal exposure to evaluate the copper stress on biomass, cellular and lipid contents. Preliminary growth studies indicated the growth tolerance levels of Chlorella in the presence of copper at 4.0 mg L-1 concentration. The total chlorophyll, protein and lipid content of the isolates were 1.7-3.45%, 0.43-0.70 mg g-1 and 0.02-0.11 mg g-1 respectively. Gas Chromatography-Mass Spectroscopy analysis revealed that the percent composition of fatty acids varied among the species studied and the major group of fatty acids were C16:0, C18:1 and C18:2. Highest percent of fatty acids were found in C. vulgaris, C. protothecoides and C. pyrenoidosa. Copper have an impact on Chlorella species where biomass content was directly proportional to the lipid productivity. The results reflects the fact that copper stress on Chlorella species as the evidence of lipid production in both qualitative and quantitative manner. In conclusion, Chlorella species can be used for the sustainable producion of renewable energy through copper stress and removal of copper from aqueous solutions.

  17. Phenolic contents and cellular antioxidant activity of Chinese hawthorn "Crataegus pinnatifida".

    Science.gov (United States)

    Wen, Lingrong; Guo, Xingbo; Liu, Rui Hai; You, Lijun; Abbasi, Arshad Mehmood; Fu, Xiong

    2015-11-01

    It is evident from various epidemiological studies that consumption of fruits and vegetables is essential to maintain health and in the disease prevention. Present study was designed to examine phenolic contents and antioxidant properties of three varieties of Crataegus pinnatifida (Chinese hawthorn). Shanlihong variety exhibited elevated levels of total phenolics and flavonoid contents, including free and bond phenolics. Procyanidin B2 was most abundant phenolic compound in all samples, followed by epicatechin, chlorogenic acid, hyperoside, and isoquercitrin. The free ORAC values, and free hydro-PSC values were 398.3-555.8 μmol TE/g DW, and 299.1-370.9 μmol VCE/g DW, respectively. Moreover, the free cellular antioxidant activity (CAA) values were 678-1200 μmol of QE/100 g DW in the no PBS wash protocol, and 345.9-532.9 μmol of QE/100 g DW in the PBS wash protocol. C. pinnatifida fruit could be valuable to promote consumer health. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Copper oxide nanoparticles stimulate glycolytic flux and increase the cellular contents of glutathione and metallothioneins in cultured astrocytes.

    Science.gov (United States)

    Bulcke, Felix; Dringen, Ralf

    2015-01-01

    Copper oxide nanoparticles (CuO-NPs) are frequently used for industrial or medical applications and are known for their high toxic potential. As little is known so far on the consequences of an exposure of brain cells to such particles, we applied CuO-NPs to cultured primary rat astrocytes and investigated whether such particles affect cell viability and alter their metabolic properties. Astrocytes efficiently accumulated CuO-NPs in a time- and concentration-dependent manner. The cells remained viable during a 24 h incubation with 100 µM copper in the form of CuO-NPs, while higher concentrations of CuO-NPs severely compromised the cell viability. Astrocytes that were exposed for 24 h to 100 µM CuO-NPs showed significantly enhanced extracellular lactate concentrations and increased cellular levels of glutathione and metallothioneins. The CuO-NP-induced increase in lactate release and metallothionein content were prevented by the presence of the membrane-permeable copper chelator tetrathiomolybdate, while this chelator increased already in the absence of CuO-NPs the cellular glutathione content. After removal of the CuO-NPs following a 24 h pre-incubation with 100 µM CuO-NPs, astrocytes maintained during a further 6 h incubation an elevated glycolytic lactate release and exported significantly more glutathione than control cells that had been pre-incubated without CuO-NPs. These data suggest that copper ions which are liberated from internalized CuO-NPs stimulate glycolytic flux as well as the synthesis of glutathione and metallothioneins in cultured viable astrocytes.

  19. New aluminium alloys with high lithium content

    Energy Technology Data Exchange (ETDEWEB)

    Schemme, K.; Velten, B.

    1989-06-01

    Since the early 80's there have been made great efforts to replace the high strength aluminium alloys for the aircraft and space industry by a new generation of aluminium-lithium alloys. The attractivity of this kind of alloys could be increased by a further reduction of their density, caused by an increasing lithium content (/ge/ 5 wt.% Li). Therefore binary high-lithium containing alloys with low density are produced and metallografically investigated. A survey of their strength and wear behavior is given by using tensile tests and pin abrasing tests. (orig.).

  20. Assessing the implications of cellular network performance on mobile content access

    DEFF Research Database (Denmark)

    Kaup, Fabian; Michelinakis, Foivos; Bui, Nicola

    2016-01-01

    that the association of mobile devices to a point of presence (PoP) within the operator's network can influence the end-to-end performance by a large extent. Given the collected data, a model predicting the PoP assignment and its resulting RTT leveraging Markov chain and machine learning approaches is developed. RTT......Mobile applications such as VoIP, (live) gaming, or video streaming have diverse QoS requirements ranging from low delay to high throughput. The optimization of the network quality experienced by end-users requires detailed knowledge of the expected network performance. Also, the achieved service...... quality is affected by a number of factors, including network operator and available technologies. However, most studies measuring the cellular network do not consider the performance implications of network configuration and management. To this end, this paper reports about an extensive data set...

  1. Information management for high content live cell imaging

    Directory of Open Access Journals (Sweden)

    White Michael RH

    2009-07-01

    Full Text Available Abstract Background High content live cell imaging experiments are able to track the cellular localisation of labelled proteins in multiple live cells over a time course. Experiments using high content live cell imaging will generate multiple large datasets that are often stored in an ad-hoc manner. This hinders identification of previously gathered data that may be relevant to current analyses. Whilst solutions exist for managing image data, they are primarily concerned with storage and retrieval of the images themselves and not the data derived from the images. There is therefore a requirement for an information management solution that facilitates the indexing of experimental metadata and results of high content live cell imaging experiments. Results We have designed and implemented a data model and information management solution for the data gathered through high content live cell imaging experiments. Many of the experiments to be stored measure the translocation of fluorescently labelled proteins from cytoplasm to nucleus in individual cells. The functionality of this database has been enhanced by the addition of an algorithm that automatically annotates results of these experiments with the timings of translocations and periods of any oscillatory translocations as they are uploaded to the repository. Testing has shown the algorithm to perform well with a variety of previously unseen data. Conclusion Our repository is a fully functional example of how high throughput imaging data may be effectively indexed and managed to address the requirements of end users. By implementing the automated analysis of experimental results, we have provided a clear impetus for individuals to ensure that their data forms part of that which is stored in the repository. Although focused on imaging, the solution provided is sufficiently generic to be applied to other functional proteomics and genomics experiments. The software is available from: fhttp://code.google.com/p/livecellim/

  2. Computer vision for high content screening.

    Science.gov (United States)

    Kraus, Oren Z; Frey, Brendan J

    2016-01-01

    High Content Screening (HCS) technologies that combine automated fluorescence microscopy with high throughput biotechnology have become powerful systems for studying cell biology and drug screening. These systems can produce more than 100 000 images per day, making their success dependent on automated image analysis. In this review, we describe the steps involved in quantifying microscopy images and different approaches for each step. Typically, individual cells are segmented from the background using a segmentation algorithm. Each cell is then quantified by extracting numerical features, such as area and intensity measurements. As these feature representations are typically high dimensional (>500), modern machine learning algorithms are used to classify, cluster and visualize cells in HCS experiments. Machine learning algorithms that learn feature representations, in addition to the classification or clustering task, have recently advanced the state of the art on several benchmarking tasks in the computer vision community. These techniques have also recently been applied to HCS image analysis.

  3. Automation in high-content flow cytometry screening.

    Science.gov (United States)

    Naumann, U; Wand, M P

    2009-09-01

    High-content flow cytometric screening (FC-HCS) is a 21st Century technology that combines robotic fluid handling, flow cytometric instrumentation, and bioinformatics software, so that relatively large numbers of flow cytometric samples can be processed and analysed in a short period of time. We revisit a recent application of FC-HCS to the problem of cellular signature definition for acute graft-versus-host-disease. Our focus is on automation of the data processing steps using recent advances in statistical methodology. We demonstrate that effective results, on par with those obtained via manual processing, can be achieved using our automatic techniques. Such automation of FC-HCS has the potential to drastically improve diagnosis and biomarker identification.

  4. High-content analysis for drug delivery and nanoparticle applications.

    Science.gov (United States)

    Brayden, David J; Cryan, Sally-Ann; Dawson, Kenneth A; O'Brien, Peter J; Simpson, Jeremy C

    2015-08-01

    High-content analysis (HCA) provides quantitative multiparametric cellular fluorescence data. From its origins in discovery toxicology, it is now addressing fundamental questions in drug delivery. Nanoparticles (NPs), polymers, and intestinal permeation enhancers are being harnessed in drug delivery systems to modulate plasma membrane properties and the intracellular environment. Identifying comparative mechanistic cytotoxicity on sublethal events is crucial to expedite the development of such systems. NP uptake and intracellular routing pathways are also being dissected using chemical and genetic perturbations, with the potential to assess the intracellular fate of targeted and untargeted particles in vitro. As we discuss here, HCA is set to make a major impact in preclinical delivery research by elucidating the intracellular pathways of NPs and the in vitro mechanistic-based toxicology of formulation constituents.

  5. High content screening in microfluidic devices

    Science.gov (United States)

    Cheong, Raymond; Paliwal, Saurabh; Levchenko, Andre

    2011-01-01

    Importance of the field Miniaturization is key to advancing the state-of-the-art in high content screening (HCS), in order to enable dramatic cost savings through reduced usage of expensive biochemical reagents and to enable large-scale screening on primary cells. Microfluidic technology offers the potential to enable HCS to be performed with an unprecedented degree of miniaturization. Areas covered in this review This perspective highlights a real-world example from the authors’ work of HCS assays implemented in a highly miniaturized microfluidic format. Advantages of this technology are discussed, including cost savings, high throughput screening on primary cells, improved accuracy, the ability to study complex time-varying stimuli, and ease of automation, integration, and scaling. What the reader will gain The reader will understand the capabilities of a new microfluidics-based platform for HCS, and the advantages it provides over conventional plate-based HCS. Take home message Microfluidics technology will drive significant advancements and broader usage and applicability of HCS in drug discovery. PMID:21852997

  6. Effect of X irradiation on a heterotransplanted human colonic carcinoma before and after a change in the cellular DNA content

    DEFF Research Database (Denmark)

    Spang-Thomsen, M; Vindeløv, L L; Nielsen, A

    1983-01-01

    A spontaneous change of cellular DNA content occurred in a hyperdiploid human colonic carcinoma grown in nude mice. After the change to hyperpentaploidy the tumor was exposed to single-dose X irradiation, and the effects on growth curves and on the cell cycle, determined by flow cytometric DNA...... analysis (FCM), were compared to results obtained with the tumor prior to the evolutionary event. The results showed that the radiation effects on growth rate and on cell kinetics had changed after the change in cellular DNA content. In the hyperpentaploid tumor the irradiation had no effect...... a partial synchronization of accumulated cells, whereas no synchronization effect was found in the hyperdiploid tumor. The redistribution time was 8-10 days for both tumors. The results indicate that clonal evolution may affect radiosensitivity, and that FCM analysis may prove to be a valuable method...

  7. Community structure, cellular rRNA content, and activity of sulfate-reducing bacteria in marine Arctic sediments

    DEFF Research Database (Denmark)

    Ravenschlag, K.; Sahm, K.; Knoblauch, C.;

    2000-01-01

    The community structure of sulfate-reducing bacteria (SRB) of a marine Arctic sediment (Smeerenburg-fjorden, Svalbard) a-as characterized by both fluorescence in situ hybridization (FISH) and rRNA slot blot hybridization by using group- and genus-specific 16S rRNA-targeted oligonucleotide probes...... that FISH and rRNA slot blot hybridization gave comparable results. Furthermore, a combination of the two methods allowed us to calculate specific cellular rRNA contents with respect to localization in the sediment profile. The rRNA contents of Desulfosarcina-Desulfococcus cells were highest in the first 5...... mm of the sediment (0.9 and 1.4 fg, respectively) and decreased steeply with depth, indicating that maximal metabolic activity occurred close to the surface, Based on SRB cell numbers, cellular sulfate reduction rates were calculated. The rates were highest in the surface layer (0.14 fmol cell(-1...

  8. Proteomic Analysis of the Marine Cyanobacterium Synechococcus WH8102 and Implications for Estimates of the Cellular Iron Content

    Science.gov (United States)

    Saito, M. A.; Bertrand, E. M.; Bulygin, V.; Moran, D.; Waterbury, J. B.

    2008-12-01

    The proteome of the marine cyanobacterium Synechococcus WH8102 was analyzed by nanospray liquid chromatography mass spectrometry (nLC-MS) with two major goals: to provide a first examination of the relative abundance of the most abundant proteins in this important microbe and to provide the necessary mass spectra for future quantification of biogeochemically significant proteins. Analyses of 37 nLC-MS runs of whole cell tryptic digestions and SDS-PAGE gel separated tryptic digestions resulted in a total of 636 proteins identified, 376 identified with two or more tryptic peptides. The identifications used the Sequest algorithm with stringent data filters on 54003 observed peptides, 3066 of which were unique, with a false positive rate of 2.2%. These measured proteins represent ~ 25.2% (14.8% with >= 2 peptides) of the open reading frames (ORFs) in the genome, similar to or higher than the percentage found in other cyanobacterial proteome studies thus far. The relative abundance of the more abundant proteins in the proteome was examined using the exponentially modified protein abundance index from a single nLC-MS run that identified 372 proteins (14.7% of the ORFs) from 7743 observed peptides (1224 unique peptides). Estimates of the relative abundance showed the photosynthesis and respiration category contributing approximately 32% of the total detected protein, hypothetical proteins contributing about 16%, and translation about 12%. Of biogeochemical interest, multiple types of nitrogen assimilation systems were observed to be simultaneously expressed as proteins, only 5 of the 21 B12 biosynthesis proteins were identified likely due to low abundance, and the metalloproteins metallothionein and nickel superoxide dismutase were relatively abundant. In contrast to previous predictions of a high photosystem I: photosystem II ratio of approximately 3 in the cyanobacteria and a resultant high cellular iron content, the ratio of the average relative abundances of all

  9. Durum Wheat Roots Adapt to Salinity Remodeling the Cellular Content of Nitrogen Metabolites and Sucrose

    Science.gov (United States)

    Annunziata, Maria Grazia; Ciarmiello, Loredana F.; Woodrow, Pasqualina; Maximova, Eugenia; Fuggi, Amodio; Carillo, Petronia

    2017-01-01

    Plants are currently experiencing increasing salinity problems due to irrigation with brackish water. Moreover, in fields, roots can grow in soils which show spatial variation in water content and salt concentration, also because of the type of irrigation. Salinity impairs crop growth and productivity by inhibiting many physiological and metabolic processes, in particular nitrate uptake, translocation, and assimilation. Salinity determines an increase of sap osmolality from about 305 mOsmol kg−1 in control roots to about 530 mOsmol kg−1 in roots under salinity. Root cells adapt to salinity by sequestering sodium in the vacuole, as a cheap osmoticum, and showing a rearrangement of few nitrogen-containing metabolites and sucrose in the cytosol, both for osmotic adjustment and oxidative stress protection, thus providing plant viability even at low nitrate levels. Mainly glycine betaine and sucrose at low nitrate concentration, and glycine betaine, asparagine and proline at high nitrate levels can be assumed responsible for the osmotic adjustment of the cytosol, the assimilation of the excess of ammonium and the scavenging of ROS under salinity. High nitrate plants with half of the root system under salinity accumulate proline and glutamine in both control and salt stressed split roots, revealing that osmotic adjustment is not a regional effect in plants. The expression level and enzymatic activities of asparagine synthetase and Δ1-pyrroline-5-carboxylate synthetase, as well as other enzymatic activities of nitrogen and carbon metabolism, are analyzed. PMID:28119716

  10. [Effect of synchronous perfusion of NaN3 in changes in content of cholinergic neurotransmitter in medial prefrontal cortex and hippocampal extra-cellular fluid].

    Science.gov (United States)

    Zhang, Mei-Yu; Sun, Dan-Dan; Liu, Yang; Cui, Yue; Zhao, Xiao-Liang; Zhang, Ying; Wang, Zhi-Guo; Wang, Dan-Qiao

    2014-02-01

    To observe the effect of synchronous perfusion of specific respiratory chain complex IV inhibitor sodium azide (NaN3) in brain on rat ventromedial prefrontal cortex (mPFC) and acetylcholine (ACh) and choline (Ch) contents in hippocampal extra-cellular fluid, and establish the AD rat model induced by mitochondrial acute injury. The synchronous dual-probe dual-channel brain microdialysis sampling technology was applied to synchronously perfuse modified Ringer's solution containing NaN3 (50 micro mol L-1) and neostigmine (2 micro mol L-1) into mPFC and hippocampus of conscious, freely moving normal rats, and continuously collect dialysates from different encephalic areas. Dynamic contents of ACh and Ch were determined by high performance liquid chromatography-post-column immobilized enzyme reactor-electrochemical process. ACh and Ch contents in mPFC extracellular fluid of normal rats were higher than that in hippocampus. During the process of perfusion, NaN3 could significantly reduce ACh in mPFC/hippocampal extra-cellular fluid, but remarkably increase Ch, and constantly inhibit the recovery of ACh and Ch contents in mPFC/hippocampus. The synchronous perfusion of NaN3in rat mPFC and hippocampus can injure functions of the cholinergic nerve projection area, and cause the acute AD model with ACh and Ch metabolic disorders. This model can be used in pathogenetic and pharmacological studies.

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

    Directory of Open Access Journals (Sweden)

    Gisela M F Vaz

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

  12. Pungency in paprika (Capsicum annuum). 1. Decrease of capsaicinoid content following cellular disruption.

    Science.gov (United States)

    Kirschbaum-Titze, Petra; Hiepler, Constanze; Mueller-Seitz, Erika; Petz, Michael

    2002-02-27

    The capsaicinoid content in fruits of Capsicum annuum decreased within several days to a level of only 10% of the starting value when cells were disrupted by homogenization. This decrease was not observed in fruits that were carefully cut into halves. The analysis of one half made it possible to determine the reference content at time zero for the second half. A much lower decrease was observed when minced fruits were stored under nitrogen, whereas storage under oxygen resulted in considerable losses of capsaicinoids, indicating oxidative processes as a cause for the decrease of capsaicinoid content.

  13. High-content single-cell analysis on-chip using a laser microarray scanner.

    Science.gov (United States)

    Zhou, Jing; Wu, Yu; Lee, Sang-Kwon; Fan, Rong

    2012-12-07

    High-content cellomic analysis is a powerful tool for rapid screening of cellular responses to extracellular cues and examination of intracellular signal transduction pathways at the single-cell level. In conjunction with microfluidics technology that provides unique advantages in sample processing and precise control of fluid delivery, it holds great potential to transform lab-on-a-chip systems for high-throughput cellular analysis. However, high-content imaging instruments are expensive, sophisticated, and not readily accessible. Herein, we report on a laser scanning cytometry approach that exploits a bench-top microarray scanner as an end-point reader to perform rapid and automated fluorescence imaging of cells cultured on a chip. Using high-content imaging analysis algorithms, we demonstrated multiplexed measurements of morphometric and proteomic parameters from all single cells. Our approach shows the improvement of both sensitivity and dynamic range by two orders of magnitude as compared to conventional epifluorescence microscopy. We applied this technology to high-throughput analysis of mesenchymal stem cells on an extracellular matrix protein array and characterization of heterotypic cell populations. This work demonstrates the feasibility of a laser microarray scanner for high-content cellomic analysis and opens up new opportunities to conduct informative cellular analysis and cell-based screening in the lab-on-a-chip systems.

  14. Evolutionary Computational Method of Facial Expression Analysis for Content-based Video Retrieval using 2-Dimensional Cellular Automata

    CERN Document Server

    Geetha, P

    2010-01-01

    In this paper, Deterministic Cellular Automata (DCA) based video shot classification and retrieval is proposed. The deterministic 2D Cellular automata model captures the human facial expressions, both spontaneous and posed. The determinism stems from the fact that the facial muscle actions are standardized by the encodings of Facial Action Coding System (FACS) and Action Units (AUs). Based on these encodings, we generate the set of evolutionary update rules of the DCA for each facial expression. We consider a Person-Independent Facial Expression Space (PIFES) to analyze the facial expressions based on Partitioned 2D-Cellular Automata which capture the dynamics of facial expressions and classify the shots based on it. Target video shot is retrieved by comparing the similar expression is obtained for the query frame's face with respect to the key faces expressions in the database video. Consecutive key face expressions in the database that are highly similar to the query frame's face, then the key faces are use...

  15. Modification of Cellular Cholesterol Content Affects Traction Force, Adhesion and Cell Spreading.

    Science.gov (United States)

    Norman, Leann L; Oetama, Ratna J; Dembo, Micah; Byfield, F; Hammer, Daniel A; Levitan, Irena; Aranda-Espinoza, Helim

    2010-06-01

    Cellular cholesterol is a critical component of the plasma membrane, and plays a key role in determining the physical properties of the lipid bilayer, such as elasticity, viscosity, and permeability. Surprisingly, it has been shown that cholesterol depletion increases cell stiffness, not due to plasma membrane stiffening, but rather, due to the interaction between the actin cytoskeleton and the plasma membrane. This indicates that traction stresses of the acto-myosin complex likely increase during cholesterol depletion. Here we use force traction microscopy to quantify the forces individual cells are exerting on the substrate, and total internal reflection fluorescence microscopy as well as interference reflection microscopy to observe cell-substrate adhesion and spreading. We show that single cells depleted of cholesterol produce larger traction forces and have large focal adhesions compared to untreated or cholesterol-enriched cells. Cholesterol depletion also causes a decrease in adhesion area for both single cells and monolayers. Spreading experiments illustrate a decrease in spreading area for cholesterol-depleted cells, and no effect on cholesterol-enriched cells. These results demonstrate that cholesterol plays an important role in controlling and regulating the cell-substrate interactions through the actin-plasma membrane complex, cell-cell adhesion, and spreading.

  16. Network-Coded Content Delivery in Femtocaching-Assisted Cellular Networks

    KAUST Repository

    Shnaiwer, Yousef N.

    2015-12-06

    Next-generation cellular networks are expected to be assisted by femtocaches (FCs), which collectively store the most popular files for the clients. Given any arbitrary non-fragmented placement of such files, a strict no-latency constraint, and clients\\' prior knowledge, new file download requests could be efficiently handled by both the FCs and the macrocell base station (MBS) using opportunistic network coding (ONC). In this paper, we aim to find the best allocation of coded file downloads to the FCs so as to minimize the MBS involvement in this download process. We first formulate this optimization problem over an ONC graph, and show that it is NP-hard. We then propose a greedy approach that maximizes the number of files downloaded by the FCs, with the goal to reduce the download share of the MBS. This allocation is performed using a dual conflict ONC graph to avoid conflicts among the FC downloads. Simulations show that our proposed scheme almost achieves the optimal performance and significantly saves on the MBS bandwidth.

  17. Cast Iron With High Carbon Content

    Science.gov (United States)

    Curreri, P. A.; Hendrix, J. C.; Stefanescu, D. M.

    1986-01-01

    Method proposed for solidifying high-carbon cast iron without carbon particles segregating at upper surface. Solidification carried out in low gravity, for example on airplane flying free-fall parabolic trajectory. Many different microstructures obtained by proposed technique, and percentage by weight of carbon retained in melt much higher than at present.

  18. Quantitative characterization of mitosis-blocked tetraploid cells using high content analysis.

    Science.gov (United States)

    Grove, Linnette E; Ghosh, Richik N

    2006-08-01

    A range of cellular evidence supporting a G1 tetraploidy checkpoint was obtained from different assay methods including flow cytometry, immunoblotting, and microscopy. Cancer research would benefit if these cellular properties could instead be measured by a single, quantitative, automated assay method, such as high content analysis (HCA). Thus, nocodazole-treated cells were fluorescently labeled for different cell cycle-associated properties, including DNA content, retinoblastoma (Rb) and histone H3 phosphorylation, p53 and p21(WAF1) expression, nuclear and cell sizes, and cell morphology, and automatically imaged, analyzed, and correlated using HCA. HCA verified that nocodazole-induced mitosis block resulted in tetraploid cells. Rb and histone H3 were maximally hyperphosphorylated by 24 h of nocodazole treatment, accompanied by cell and nuclear size decreases and cellular rounding. Cells remained tetraploid and mononucleated with longer treatments, but other targets reverted to G1 levels, including Rb and histone H3 dephosphorylation accompanied by cellular respreading. This was accompanied by increased p53 and p21(WAF1) expression levels. The range of effects accompanying nocodazole-induced block of mitosis and the resulting tetraploid cells' reversal to a pseudo-G1 state can be quantitatively measured by HCA in an automated manner, recommending this assay method for the large-scale biology challenges of modern cancer drug discovery.

  19. A Study of cellular phone possessors' sex consciousness and sex action in high school

    OpenAIRE

    池田, かよ子; 久保田, 美雪; 渡邊, 典子; Ikeda, Kayoko; Kubota, Miyuki; Watanabe, Noriko

    2004-01-01

    The purpose of this survey for 304 high school students (116 boys and 188 girls) is to make sure how high school students who possess cellular phones use their phones, what they use their phones for, to what extent they get information about sex, how far they feel conscious of sex and their sex action. (1) About 60% of the girls have cellular phones, which is larger than the rate of the boys who have cellularphones. (2) As for the frequency of using cellular phones, about 80% of the girls use...

  20. Cognitive radio networks efficient resource allocation in cooperative sensing, cellular communications, high-speed vehicles, and smart grid

    CERN Document Server

    Jiang, Tao; Cao, Yang

    2015-01-01

    PrefaceAcknowledgmentsAbout the AuthorsIntroductionCognitive Radio-Based NetworksOpportunistic Spectrum Access NetworksCognitive Radio Networks with Cooperative SensingCognitive Radio Networks for Cellular CommunicationsCognitive Radio Networks for High-Speed VehiclesCognitive Radio Networks for a Smart GridContent and OrganizationTransmission Slot Allocation in an Opportunistic Spectrum Access NetworkSingle-User Single-Channel System ModelProbabilistic Slot Allocation SchemeOptimal Probabilistic Slot AllocationBaseline PerformanceExponential DistributionHyper-Erlang DistributionPerformance An

  1. High-content imaging with micropatterned multiwell plates reveals influence of cell geometry and cytoskeleton on chromatin dynamics.

    Science.gov (United States)

    Harkness, Ty; McNulty, Jason D; Prestil, Ryan; Seymour, Stephanie K; Klann, Tyler; Murrell, Michael; Ashton, Randolph S; Saha, Krishanu

    2015-10-01

    Understanding the mechanisms underpinning cellular responses to microenvironmental cues requires tight control not only of the complex milieu of soluble signaling factors, extracellular matrix (ECM) connections and cell-cell contacts within cell culture, but also of the biophysics of human cells. Advances in biomaterial fabrication technologies have recently facilitated detailed examination of cellular biophysics and revealed that constraints on cell geometry arising from the cellular microenvironment influence a wide variety of human cell behaviors. Here, we create an in vitro platform capable of precise and independent control of biochemical and biophysical microenvironmental cues by adapting microcontact printing technology into the format of standard six- to 96-well plates to create MicroContact Printed Well Plates (μCP Well Plates). Automated high-content imaging of human cells seeded on μCP Well Plates revealed tight, highly consistent control of single-cell geometry, cytoskeletal organization, and nuclear elongation. Detailed subcellular imaging of the actin cytoskeleton and chromatin within live human fibroblasts on μCP Well Plates was then used to describe a new relationship between cellular geometry and chromatin dynamics. In summary, the μCP Well Plate platform is an enabling high-content screening technology for human cell biology and cellular engineering efforts that seek to identify key biochemical and biophysical cues in the cellular microenvironment.

  2. The aluminium content of infant formulas remains too high

    OpenAIRE

    2013-01-01

    Background Recent research published in this journal highlighted the issue of the high content of aluminium in infant formulas. The expectation was that the findings would serve as a catalyst for manufacturers to address a significant problem of these, often necessary, components of infant nutrition. It is critically important that parents and other users have confidence in the safety of infant formulas and that they have reliable information to use in choosing a product with a lower content ...

  3. Establishing a High-content Analysis Method for Tubulin Polymerization to Evaluate Both the Stabilizing and Destabilizing Activities of Compounds.

    Science.gov (United States)

    Sum, Chi Shing; Nickischer, Debra; Lei, Ming; Weston, Andrea; Zhang, Litao; Schweizer, Liang

    2014-01-01

    Microtubules are important components of the cellular cytoskeleton that play roles in various cellular processes such as vesicular transport and spindle formation during mitosis. They are formed by an ordered organization of α-tubulin and β-tubulin hetero-polymers. Altering microtubule polymerization has been known to be the mechanism of action for a number of therapeutically important drugs including taxanes and epothilones. Traditional cell-based assays for tubulin-interacting compounds rely on their indirect effects on cell cycle and/or cell proliferation. Direct monitoring of compound effects on microtubules is required to dissect detailed mechanisms of action in a cellular setting. Here we report a high-content assay platform to monitor tubulin polymerization status by directly measuring the acute effects of drug candidates on the cellular tubulin network with the capability to dissect the mechanisms of action. This high-content analysis distinguishes in a quantitative manner between compounds that act as tubulin stabilizers versus those that are tubulin destabilizers. In addition, using a multiplex approach, we expanded this analysis to simultaneously monitor physiological cellular responses and associated cellular phenotypes.

  4. Phase-field simulation of formation of cellular dendrites and fine cellular structures at high growth velocities during directional solidification of Ti56Al44 alloy

    Institute of Scientific and Technical Information of China (English)

    LI Xin-zhong; GUO Jing-jie; SU Yan-qing; WU Shi-ping; FU Heng-zhi

    2005-01-01

    A phase-field model whose free energy of the solidification system derived from the Calphad thermodynamic modeling of phase diagram was used to simulate formation of cellular dendrites and fine cellular structures of Ti56Al44 alloy during directional solidification at high growth velocities. The liquid-solid phase transition of L→β was chosen. The dynamics of breakdown of initially planar interfaces into cellular dendrites and fine cellular structures were shown firstly at two growth velocities. Then the unidirectional free growths of two initial nucleations evolving to fine cellular dendrites were investigated. The tip splitting phenomenon is observed and the negative temperature gradient in the liquid represents its supercooling directional solidification. The simulation results show the realistic evolution of interfaces and microstructures and they agree with experimental one.

  5. Effects of the breed, sex and age on cellular content and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel

    OpenAIRE

    Giraldo Carlos E; López Catalina; Álvarez María E; Samudio Ismael J; Prades Marta; Carmona Jorge U

    2013-01-01

    Abstract Background There is no information on the effects of the breed, gender and age on the cellular content and growth factor (GF) release from equine pure-platelet rich plasma (P-PRP) and pure-platelet rich gel (P-PRG). The objectives of this study were: 1) to compare the cellular composition of P-PRP with whole blood and platelet poor plasma (PPP); 2) to compare the concentration of transforming GF beta 1 (TGF-β1) and platelet derived GF isoform BB (PDGF-BB) between P-PRP treated with n...

  6. EFFECT OF CARBON CONTENT ON MICROSTRUCTURE AND PROPERTIES OF HIGH STRENGTH AND HIGH ELONGATION STEELS

    Institute of Scientific and Technical Information of China (English)

    Y.Chen; X.Chen; 等

    2003-01-01

    The microstructure and mechanical properties of new kind of hot-rolled high strength and high elongation steels with retained austenite were studied by discussing the in-fluence of different carbon content.The research results indicate that carbon content has a significant effect on retaining austenite and consequently resulting in high elon-gation.Besides,new findings about relationship between carbon content and retained austenite as well as properties were discussed in the paper.

  7. Evaluation of dissolution rate on high plutonium content MOX fuel

    Energy Technology Data Exchange (ETDEWEB)

    Sugaya, Shinichi; Kurita, Ichiro; Endo, Hideo; Higuchi, Hidetoshi; Kihara, Yoshiyuki [Japan Nuclear Cycle Development Inst., Tokai Works, Tokai, Ibaraki (Japan); Ogasawara, Masahiro; Shinada, Masanori; Kowata, Masato [Inspection Development Company Ltd., Tokai, Ibaraki (Japan)

    2002-06-01

    The dissolution rate of high Pu content MOX fuel into nitric acid was measured as a function of Pu content. MOX fuel samples, pressed and sintered, were dissolved in 7 M of boiling nitric acid, and the dissolution rate was measured by analyzing the Pu and U concentration in the solution. The dissolution rate of MOX fuel tended to decrease with the increase in the Pu content and was reduced after 6 hours of dissolution. These results agreed well with previous ones, but the dissolution rate was 3-6 times faster than those. It is estimated that the cause of this difference was due to underestimation of the surface area of MOX fuel powder and the difference of the MOX O/M ratio. (author)

  8. Developmental toxicity assay using high content screening of zebrafish embryos.

    Science.gov (United States)

    Lantz-McPeak, Susan; Guo, Xiaoqing; Cuevas, Elvis; Dumas, Melanie; Newport, Glenn D; Ali, Syed F; Paule, Merle G; Kanungo, Jyotshna

    2015-03-01

    Typically, time-consuming standard toxicological assays using the zebrafish (Danio rerio) embryo model evaluate mortality and teratogenicity after exposure during the first 2 days post-fertilization. Here we describe an automated image-based high content screening (HCS) assay to identify the teratogenic/embryotoxic potential of compounds in zebrafish embryos in vivo. Automated image acquisition was performed using a high content microscope system. Further automated analysis of embryo length, as a statistically quantifiable endpoint of toxicity, was performed on images post-acquisition. The biological effects of ethanol, nicotine, ketamine, caffeine, dimethyl sulfoxide and temperature on zebrafish embryos were assessed. This automated developmental toxicity assay, based on a growth-retardation endpoint should be suitable for evaluating the effects of potential teratogens and developmental toxicants in a high throughput manner. This approach can significantly expedite the screening of potential teratogens and developmental toxicants, thereby improving the current risk assessment process by decreasing analysis time and required resources.

  9. Cellular transformation by human papillomaviruses: Lessons learned by comparing high- and low-risk viruses

    Science.gov (United States)

    Klingelhutz, Aloysius J.; Roman, Ann

    2013-01-01

    The oncogenic potential of papillomaviruses (PVs) has been appreciated since the 1930s yet the mechanisms of virally-mediated cellular transformation are still being revealed. Reasons for this include: a) the oncoproteins are multifunctional, b) there is an ever-growing list of cellular interacting proteins, c) more than one cellular protein may bind to a given region of the oncoprotein, and d) there is only limited information on the proteins encoded by the corresponding non-oncogenic PVs. The perspective of this review will be to contrast the activities of the viral E6 and E7 proteins encoded by the oncogenic human PVs (termed high-risk HPVs) to those encoded by their non-oncogenic counterparts (termed low-risk HPVs) in an attempt to sort out viral life cycle-related functions from oncogenic functions. The review will emphasize lessons learned from the cell culture studies of the HPVs causing mucosal/genital tract cancers. PMID:22284986

  10. Using Shared Memory As A Cache In High Performance Cellular Automata Water Flow Simulations

    Directory of Open Access Journals (Sweden)

    Paweł Topa

    2013-01-01

    Full Text Available Graphics processors (GPU -- Graphic Processor Units recently have gained a lot of interest as an efficient platform for general-purpose computation. Cellular Automata approach which is inherently parallel gives the opportunity to implement high performance simulations. This paper presents how shared memory in GPU can be used to improve performance for Cellular Automata models. In our previous works, we proposed algorithms for Cellular Automata model that use only a GPU global memory. Using a profiling tool, we found bottlenecks in our approach. We introduce modifications that takes an advantage of fast shared memory. The modified algorithm is presented in details, and the results of profiling and performance test are demonstrated. Our unique achievement is comparing the efficiency of the same algorithm working with a global and shared memory.

  11. Separation/enrichment of the low-content high molecular weight natural protein using protein-imprinted polymers with ARPCs

    Institute of Scientific and Technical Information of China (English)

    XIA JianJun; LONG Yi; GUO MinJie; WANG Ying; MI HuaiFeng

    2009-01-01

    We introduce a new method for separation/enrichment of the low-content cellular protein in high molecular weight on the basis of molecular imprinting.The template protein,bacterial cloned immunoglobulin binding protein (BiP),was selectively assembled with assistant recognition polymer chains (ARPCs) from their library,which consists of numerous limited length polymer chains with randomly distributed recognition and immobilizing sites.The assemblies of proteins and ARPCs were adsorbed by porous polymeric beads and immobilized by cross-linking polymerization.After the template was removed,the synthesized imprinted polymer was used to adsorb authentic BiP from endoplasmic reticulum (ER) extract,and its proportional content was enriched 45 times.It is the first time that the low-content cellular natural protein,whose molecular weight reaches 78 kDa,is enrichd by molecular imprinting.

  12. Separation/enrichment of the low-content high molecular weight natural protein using protein-imprinted polymers with ARPCs

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    We introduce a new method for separation/enrichment of the low-content cellular protein in high mo-lecular weight on the basis of molecular imprinting. The template protein, bacterial cloned immu-noglobulin binding protein (BiP), was selectively assembled with assistant recognition polymer chains (ARPCs) from their library, which consists of numerous limited length polymer chains with randomly distributed recognition and immobilizing sites. The assemblies of proteins and ARPCs were adsorbed by porous polymeric beads and immobilized by cross-linking polymerization. After the template was removed, the synthesized imprinted polymer was used to adsorb authentic BiP from endoplasmic re-ticulum (ER) extract, and its proportional content was enriched 45 times. It is the first time that the low-content cellular natural protein, whose molecular weight reaches 78 kDa, is enriched by molecular imprinting.

  13. Molecular Dynamics of Materials Possessing High Energy Content.

    Science.gov (United States)

    1988-01-26

    I -RI90 634 MOLECULAR DYNAMICS OF MATERIALS POSSESSING HIGH ENERGY 1/1 r CONTENTCU) COLUMBIA UNIV MENd YORK N J TURRO 26 JAN GO I RFOSR-TR-88-0168...Bolling Air Force Base, D.C. 2 61102F_ 2303 I B2 11 T,TL.E (Inciuoe Security Classification) Molecular Dynamics of Materials Possessing High Energy...York 10027 (212) 280-2175 TITLE: MOLECULAR DYNAMICS OF MATERIALS POSSESSING HIGH ENERGY CONTENT .. 0 0 88 2 ... "" ’% ,i u , . .. .. ....... ŝ" ;! ,i

  14. Changes in protein and nonprotein thiol contents in bladder, kidney and liver of mice by the pesticide sodium-o-phenylphenol and their possible role in cellular toxicity.

    Science.gov (United States)

    Narayan, S; Roy, D

    1992-02-01

    Acute treatment of mice with Na-o-phenylphenol or phenylbenzoquinone, an electrophilic metabolite of o-phenylphenol, resulted in differential depletion of contents of protein and nonprotein thiols in bladder, kidney and liver. Maximum decrease in the levels of protein and nonprotein reduced thiols was observed in bladder (by both agents) and was followed by kidney (by both agents) and liver (phenylbenzoquinone only). The reason for this differential changes in reduced thiol contents remains to be understood. The content of protein and nonprotein disulfides was higher in bladder of mice treated with Na-o-phenylphenol compared to that observed in untreated mice bladder. Phenyl 2,5'-p-benzoquinone mediated in vivo depletion of nonprotein and protein thiols suggests that Na-o-phenylphenol treatment may decrease in vivo thiols via the formation of phenylbenzoquinone. Increased disulfide formation is considered to represent an index of oxidative stress produced by chemical. Increases in the level of protein and nonprotein disulfides in bladder suggest as observed in this study that administration of Na-o-phenylphenol to mice produced oxidative stress in bladder. Products of redox cycling of xenobiotics are known to cause cellular toxicity via altering the homeostasis of thiol status. Therefore, it is concluded that decreases in protein thiol contents either via alkylation and/or oxidation of sulfhydryl groups of proteins and increases in disulfide contents presumably by products of redox cycling of Na-o-phenylphenol may play a role in Na-o-phenylphenol-induced cellular toxicity.

  15. Analysis of genome content evolution in pvc bacterial super-phylum: assessment of candidate genes associated with cellular organization and lifestyle.

    Science.gov (United States)

    Kamneva, Olga K; Knight, Stormy J; Liberles, David A; Ward, Naomi L

    2012-01-01

    The Planctomycetes, Verrucomicrobia, Chlamydiae (PVC) super-phylum contains bacteria with either complex cellular organization or simple cell structure; it also includes organisms of different lifestyles (pathogens, mutualists, commensal, and free-living). Genome content evolution of this group has not been studied in a systematic fashion, which would reveal genes underlying the emergence of PVC-specific phenotypes. Here, we analyzed the evolutionary dynamics of 26 PVC genomes and several outgroup species. We inferred HGT, duplications, and losses by reconciliation of 27,123 gene trees with the species phylogeny. We showed that genome expansion and contraction have driven evolution within Planctomycetes and Chlamydiae, respectively, and balanced each other in Verrucomicrobia and Lentisphaerae. We also found that for a large number of genes in PVC genomes the most similar sequences are present in Acidobacteria, suggesting past and/or current ecological interaction between organisms from these groups. We also found evidence of shared ancestry between carbohydrate degradation genes in the mucin-degrading human intestinal commensal Akkermansia muciniphila and sequences from Acidobacteria and Bacteroidetes, suggesting that glycoside hydrolases are transferred laterally between gut microbes and that the process of carbohydrate degradation is crucial for microbial survival within the human digestive system. Further, we identified a highly conserved genetic module preferentially present in compartmentalized PVC species and possibly associated with the complex cell plan in these organisms. This conserved machinery is likely to be membrane targeted and involved in electron transport, although its exact function is unknown. These genes represent good candidates for future functional studies.

  16. Aggressive content of high school students' TV viewing.

    Science.gov (United States)

    Yoon, Jina S; Somers, Cheryl L

    2003-12-01

    The purpose of this study was to examine high school students' exposure to television programming with aggressive content and to explore whether consumption of aggressive TV varied by sex and ethnicity. Participants were 472 boys and girls from two high schools, one urban and one suburban. Definitions of both direct and indirect aggression were used to rate TV programs, and the participants' exposure to both was assessed. Analysis yielded a statistically significant effect for sex but not ethnicity as girls watched more TV programs containing indirect aggression. Also, exposure to aggressive TV content peaked in Grade 10 and fell sharply thereafter. The importance of educating adolescents about the images they view is highlighted. Implications for research are discussed.

  17. Method for creating high carbon content products from biomass oil

    Science.gov (United States)

    Parker, Reginald; Seames, Wayne

    2012-12-18

    In a method for producing high carbon content products from biomass, a biomass oil is added to a cracking reactor vessel. The biomass oil is heated to a temperature ranging from about 100.degree. C. to about 800.degree. C. at a pressure ranging from about vacuum conditions to about 20,700 kPa for a time sufficient to crack the biomass oil. Tar is separated from the cracked biomass oil. The tar is heated to a temperature ranging from about 200.degree. C. to about 1500.degree. C. at a pressure ranging from about vacuum conditions to about 20,700 kPa for a time sufficient to reduce the tar to a high carbon content product containing at least about 50% carbon by weight.

  18. The aluminium content of infant formulas remains too high.

    Science.gov (United States)

    Chuchu, Nancy; Patel, Bhavini; Sebastian, Blaise; Exley, Christopher

    2013-10-08

    Recent research published in this journal highlighted the issue of the high content of aluminium in infant formulas. The expectation was that the findings would serve as a catalyst for manufacturers to address a significant problem of these, often necessary, components of infant nutrition. It is critically important that parents and other users have confidence in the safety of infant formulas and that they have reliable information to use in choosing a product with a lower content of aluminium. Herein, we have significantly extended the scope of the previous research and the aluminium content of 30 of the most widely available and often used infant formulas has been measured. Both ready-to-drink milks and milk powders were subjected to microwave digestion in the presence of 15.8 M HNO3 and 30% w/v H2O2 and the aluminium content of the digests was measured by TH GFAAS. Both ready-to-drink milks and milk powders were contaminated with aluminium. The concentration of aluminium across all milk products ranged from ca 100 to 430 μg/L. The concentration of aluminium in two soya-based milk products was 656 and 756 μg/L. The intake of aluminium from non-soya-based infant formulas varied from ca 100 to 300 μg per day. For soya-based milks it could be as high as 700 μg per day. All 30 infant formulas were contaminated with aluminium. There was no clear evidence that subsequent to the problem of aluminium being highlighted in a previous publication in this journal that contamination had been addressed and reduced. It is the opinion of the authors that regulatory and other non-voluntary methods are now required to reduce the aluminium content of infant formulas and thereby protect infants from chronic exposure to dietary aluminium.

  19. Engineering properties for high kitchen waste content municipal solid waste

    Institute of Scientific and Technical Information of China (English)

    Wu Gao; Yunmin Chen; Liangtong Zhan; Xuecheng Bian

    2015-01-01

    Engineering properties of municipal solid waste (MSW) depend largely on the waste’s initial composition and degree of degradation. MSWs in developing countries usually have a high kitchen waste content (called HKWC MSW). After comparing and analyzing the laboratory and field test results of physical composition, hydraulic properties, gas generation and gas permeability, and mechanical properties for HKWC MSW and low kitchen waste content MSW (called LKWC MSW), the following findings were obtained: (1) HKWC MSW has a higher initial water content (IWC) than LKWC MSW, but the field ca-pacities of decomposed HKWC and LKWC MSWs are similar; (2) the hydraulic conductivity and gas permeability for HKWC MSW are both an order of magnitude smaller than those for LKWC MSW; (3) compared with LKWC MSW, HKWC MSW has a higher landfill gas (LFG) generation rate but a shorter duration and a lower potential capacity; (4) the primary compression feature for decomposed HKWC MSW is similar to that of decomposed LKWC MSW, but the compression induced by degradation of HKWC MSW is greater than that of LKWC MSW; and (5) the shear strength of HKWC MSW changes significantly with time and strain. Based on the differences of engineering properties between these two kinds of MSWs, the geo-environmental issues in HKWC MSW landfills were analyzed, including high leachate production, high leachate mounds, low LFG collection efficiency, large settlement and slope stability problem, and corresponding advice for the management and design of HKWC MSW landfills was recommended.

  20. Liquid alternative diesel fuels with high hydrogen content

    Energy Technology Data Exchange (ETDEWEB)

    Hancsok, Jenoe; Varga, Zoltan; Eller, Zoltan; Poelczmann, Gyoergy [Pannonia Univ., Veszprem (Hungary). MOL Dept. of Hydrocarbon Processing; Kasza, Tamas [MOL Hungarian Oil and Gas Plc., Szazhalombatta (Hungary)

    2013-06-01

    Mobility is a keystone of the sustainable development. In the operation of the vehicles as the tools of mobility internal combustion engines, so thus Diesel engines will play a remarkable role in the next decades. Beside fossil fuels - used for power these engines - liquid alternative fuels have higher and higher importance, because of their known advantages. During the presentation the categorization possibilities based on the chronology of their development and application will be presented. The importance of fuels with high hydrogen content will be reviewed. Research and development activity in the field of such kind of fuels will be presented. During this developed catalytic systems and main performance properties of the product will be presented which were obtained in case of biogasoils produced by special hydrocracking of natural triglycerides and in case of necessity followed by isomerization; furthermore in case of synthetic biogasoils obtained by the isomerization hydrocracking of Fischer-Tropsch paraffins produced from biomass based synthesis gas. Excellent combustion properties (cetane number > 65-75), good cold flow properties and reduced harmful material emission due to the high hydrogen content (C{sub n}H{sub 2n+2}) are highlighted. Finally production possibilities of linear and branched paraffins based on lignocelluloses are briefly reviewed. Summarizing it was concluded that liquid hydrocarbons with high isoparaffin content are the most suitable fuels regarding availability, economical and environmental aspects, namely the sustainable development. (orig.)

  1. Engineering properties for high kitchen waste content municipal solid waste

    Directory of Open Access Journals (Sweden)

    Wu Gao

    2015-12-01

    Full Text Available Engineering properties of municipal solid waste (MSW depend largely on the waste's initial composition and degree of degradation. MSWs in developing countries usually have a high kitchen waste content (called HKWC MSW. After comparing and analyzing the laboratory and field test results of physical composition, hydraulic properties, gas generation and gas permeability, and mechanical properties for HKWC MSW and low kitchen waste content MSW (called LKWC MSW, the following findings were obtained: (1 HKWC MSW has a higher initial water content (IWC than LKWC MSW, but the field capacities of decomposed HKWC and LKWC MSWs are similar; (2 the hydraulic conductivity and gas permeability for HKWC MSW are both an order of magnitude smaller than those for LKWC MSW; (3 compared with LKWC MSW, HKWC MSW has a higher landfill gas (LFG generation rate but a shorter duration and a lower potential capacity; (4 the primary compression feature for decomposed HKWC MSW is similar to that of decomposed LKWC MSW, but the compression induced by degradation of HKWC MSW is greater than that of LKWC MSW; and (5 the shear strength of HKWC MSW changes significantly with time and strain. Based on the differences of engineering properties between these two kinds of MSWs, the geo-environmental issues in HKWC MSW landfills were analyzed, including high leachate production, high leachate mounds, low LFG collection efficiency, large settlement and slope stability problem, and corresponding advice for the management and design of HKWC MSW landfills was recommended.

  2. Fibrosis in Chronic Hepatitis C: Correlation between Immunohistochemically-Assessed Virus Load with Steatosis and Cellular Iron Content

    Directory of Open Access Journals (Sweden)

    Maha Akl

    2016-11-01

    CONCLUSION: Grade of inflammation is the factor affecting fibrosis in CHC. The degree of liver damage is not related to cellular viral load or iron load. Steatosis is associated with higher inflammation and fibrosis. VEGF by IHC is correlated with overexpression of VEGFR2 by FISH.

  3. Functional genomic and high-content screening for target discovery and deconvolution

    Science.gov (United States)

    Heynen-Genel, Susanne; Pache, Lars; Chanda, Sumit K

    2014-01-01

    Introduction Functional genomic screens apply knowledge gained from the sequencing of the human genome toward rapid methods of identifying genes involved in cellular function based on a specific phenotype. This approach has been made possible through the use of advances in both molecular biology and automation. The utility of this approach has been further enhanced through the application of image-based high content screening, an automated microscopy and quantitative image analysis platform. These approaches can significantly enhance acquisition of novel targets for drug discovery. Areas covered Both the utility and potential issues associated with functional genomic screening approaches are discussed along with examples that illustrate both. The considerations for high content screening applied to functional genomics are also presented. Expert opinion Functional genomic and high content screening are extremely useful in the identification of new drug targets. However, the technical, experimental, and computational parameters have an enormous influence on the results. Thus, although new targets are identified, caution should be applied toward interpretation of screening data in isolation. Genomic screens should be viewed as an integral component of a target identification campaign that requires both the acquisition of orthogonal data, as well as a rigorous validation strategy. PMID:22860749

  4. Contents

    Directory of Open Access Journals (Sweden)

    Editor IJRED

    2012-11-01

    Full Text Available International Journal of Renewable Energy Development www.ijred.com Volume 1             Number 3            October 2012                ISSN 2252- 4940   CONTENTS OF ARTICLES page Design and Economic Analysis of a Photovoltaic System: A Case Study 65-73 C.O.C. Oko , E.O. Diemuodeke, N.F. Omunakwe, and E. Nnamdi     Development of Formaldehyde Adsorption using Modified Activated Carbon – A Review 75-80 W.D.P Rengga , M. Sudibandriyo and M. Nasikin     Process Optimization for Ethyl Ester Production in Fixed Bed Reactor Using Calcium Oxide Impregnated Palm Shell Activated Carbon (CaO/PSAC 81-86 A. Buasri , B. Ksapabutr, M. Panapoy and N. Chaiyut     Wind Resource Assessment in Abadan Airport in Iran 87-97 Mojtaba Nedaei       The Energy Processing by Power Electronics and its Impact on Power Quality 99-105 J. E. Rocha and B. W. D. C. Sanchez       First Aspect of Conventional Power System Assessment for High Wind Power Plants Penetration 107-113 A. Merzic , M. Music, and M. Rascic   Experimental Study on the Production of Karanja Oil Methyl Ester and Its Effect on Diesel Engine 115-122 N. Shrivastava,  , S.N. Varma and M. Pandey  

  5. A Microfabricated 96-Well 3D Assay Enabling High-Throughput Quantification of Cellular Invasion Capabilities

    Science.gov (United States)

    Hao, Rui; Wei, Yuanchen; Li, Chaobo; Chen, Feng; Chen, Deyong; Zhao, Xiaoting; Luan, Shaoliang; Fan, Beiyuan; Guo, Wei; Wang, Junbo; Chen, Jian

    2017-01-01

    This paper presents a 96-well microfabricated assay to study three-dimensional (3D) invasion of tumor cells. A 3D cluster of tumor cells was first generated within each well by seeding cells onto a micro-patterned surface consisting of a central fibronectin-coated area that promotes cellular attachment, surrounded by a poly ethylene glycol (PEG) coated area that is resistant to cellular attachment. Following the formation of the 3D cell clusters, a 3D collagen extracellular matrix was formed in each well by thermal-triggered gelation. Invasion of the tumor cells into the extracellular matrix was subsequently initiated and monitored. Two modes of cellular infiltration were observed: A549 cells invaded into the extracellular matrix following the surfaces previously coated with PEG molecules in a pseudo-2D manner, while H1299 cells invaded into the extracellular matrix in a truly 3D manner including multiple directions. Based on the processing of 2D microscopic images, a key parameter, namely, equivalent invasion distance (the area of invaded cells divided by the circumference of the initial cell cluster) was obtained to quantify migration capabilities of these two cell types. These results validate the feasibility of the proposed platform, which may function as a high-throughput 3D cellular invasion assay. PMID:28240272

  6. New approach to modulate retinal cellular toxic effects of high glucose using marine epa and dha

    Directory of Open Access Journals (Sweden)

    Fagon Roxane

    2011-06-01

    Full Text Available Abstract Background Protective effects of omega-3 fatty acids against cellular damages of high glucose were studied on retinal pigmented epithelial (RPE cells. Methods Retinal epithelial cells were incubated with omega-3 marine oils rich in EPA and DHA and then with high glucose (25 mM for 48 hours. Cellular responses were compared to normal glucose (5 mM: intracellular redox status, reactive oxygen species (ROS, mitochondrial succinate deshydrogenase activity, inflammatory cytokines release and caveolin-1 expression were evaluated using microplate cytometry, ELISA and flow cytometry techniques. Fatty acids incorporation in retinal cell membranes was analysed using chromatography. Results Preincubation of the cells with fish oil decreased ROS overproduction, mitochondrial alterations and TNFα release. These protective effects could be attributed to an increase in caveolin-1 expression induced by marine oil. Conclusion Marine formulations rich in omega-3 fatty acids represent a promising therapeutic approach for diabetic retinopathy.

  7. Content

    DEFF Research Database (Denmark)

    Keiding, Tina Bering

    Aim, content and methods are fundamental categories of both theoretical and practical general didactics. A quick glance in recent pedagogical literature on higher education, however, reveals a strong preoccupation with methods, i.e. how teaching should be organized socially (Biggs & Tang, 2007......; Race, 2001; Ramsden, 2003). This trend appears closely related to the ‘from-teaching-to-learning’ movement, which has had a strong influence on pedagogy since the early nineties (Keiding, 2007; Terhart, 2003). Another interpretation of the current interest in methodology can be derived from...... for selection of content (Klafki, 1985, 2000; Myhre, 1961; Nielsen, 2006). These attempts all share one feature, which is that criteria for selection of content appear very general and often, more or less explicitly, deal with teaching at the first Bologna-cycle; i.e. schooling at the primary and lower...

  8. Fruit characterization of high oil content avocado varieties

    Directory of Open Access Journals (Sweden)

    Gómez-López Vicente Manuel

    2002-01-01

    Full Text Available To expand the data bank on avocado (Persea americana Mill. varieties all over the world, and to select good varieties for commercial or improving purposes, a partial fruit characterization of 13 high oil content (11.23-18.80% was performed. The chosen varieties are growing in a Venezuelan germplasm bank: Fuerte, Peruano, Lula, Ortega, Red Collinson, Alcemio, Araira 1, Pope, Ettinger, Gripiña 5, Barker, Duke, and Ryan. They were characterized for pulp oil and moisture; weight (whole fruit, seed, pulp and peel; length, width and fruit shape; peel characteristics (roughness, color and hand peeling; and ripeness time. The variety Ryan showed the highest oil content (18.80% and calorific value (191 Kcal/100 g wet flesh. Avocado varieties grown in Venezuela have generally less oil content and are generally lighter than those from other countries. Most of the varieties had low pulp proportion, and were pyriform, with rough green peel and difficult to hand peel. Red Collinson had an uncommon reddish peel. The ripening time was between 4 and 10 days after harvest.

  9. EFFECT OF CARBON CONTENT ON MICROSTRUCTURE AND PROPERTIES OF HIGH STRENGTH AND HIGH ELONGATION STEELS

    Institute of Scientific and Technical Information of China (English)

    Y. Chen; X. Chen; P.H. Li; S.K. Pu; Z.X. Yuan; B.F. Xu; D.X. Lou; A.M. Guo; S.B.Zhou

    2003-01-01

    The microstructure and mechanical properties of new kind of hot-rolled high strengthand high elongation steels with retained austenite were studied by discussing the in-fluence of different carbon content. The research results indicate that carbon contenthas a significant effect on retaining austenite and consequently resulting in high elon-gation. Besides, new findings about relationship between carbon content and retainedaustenite as well as properties were discussed in the paper.

  10. Image analysis benchmarking methods for high-content screen design.

    Science.gov (United States)

    Fuller, C J; Straight, A F

    2010-05-01

    The recent development of complex chemical and small interfering RNA (siRNA) collections has enabled large-scale cell-based phenotypic screening. High-content and high-throughput imaging are widely used methods to record phenotypic data after chemical and small interfering RNA treatment, and numerous image processing and analysis methods have been used to quantify these phenotypes. Currently, there are no standardized methods for evaluating the effectiveness of new and existing image processing and analysis tools for an arbitrary screening problem. We generated a series of benchmarking images that represent commonly encountered variation in high-throughput screening data and used these image standards to evaluate the robustness of five different image analysis methods to changes in signal-to-noise ratio, focal plane, cell density and phenotype strength. The analysis methods that were most reliable, in the presence of experimental variation, required few cells to accurately distinguish phenotypic changes between control and experimental data sets. We conclude that by applying these simple benchmarking principles an a priori estimate of the image acquisition requirements for phenotypic analysis can be made before initiating an image-based screen. Application of this benchmarking methodology provides a mechanism to significantly reduce data acquisition and analysis burdens and to improve data quality and information content.

  11. High resolution DNA content measurements of mammalian sperm

    Energy Technology Data Exchange (ETDEWEB)

    Pinkel, D.; Lake, S.; Gledhill, B.L.; Van Dilla, M.A.; Stephenson, D.; Watchmaker, G.

    1982-01-01

    The high condensation and flat shape of the mammalian sperm nucleus present unique difficulties to flow cytometric measurement of DNA content. Chromatin compactness makes quantitative fluorescent staining for DNA difficult and causes a high index of refraction. The refractive index makes optical measurements sensitive to sperm head orientation. We demonstrate that the optical problems can be overcome using the commercial ICP22 epiillumination flow cytometer (Ortho Instruments, Westwood, MA) or a specially built cell orientating flow cytometer (OFCM). The design and operation of the OFCM are described. Measurements of the angular dependence of fluorescence from acriflavine stained rabbit sperm show that it is capable of orienting flat sperm with a tolerance of +-7/sup 0/. Differences in the angular dependence for the similarly shaped bull and rabbit sperm allow discrimination of these cells. We show that DNA staining with 4-6 diamidino-2-phenylindole (DAPI) or an ethidium bromide mithramycin combination allows resolution of the X and Y populations in mouse sperm. They have also been successful with sperm from the bull, ram, rabbit, and boar. Reliable results with human sperm are not obtained. The accuracy of the staining and measurement techniques are verified by the correct determination of the relative content of these two populations in sperm from normal mice and those with the Cattanach (7 to X) translocation. Among the potential uses of these techniques are measurement of DNA content errors induced in sperm due to mutagen exposure, and assessment of the fractions of X and Y sperm in semen that may have one population artifically enriched.

  12. Strong and moldable cellulose magnets with high ferrite nanoparticle content.

    Science.gov (United States)

    Galland, Sylvain; Andersson, Richard L; Ström, Valter; Olsson, Richard T; Berglund, Lars A

    2014-11-26

    A major limitation in the development of highly functional hybrid nanocomposites is brittleness and low tensile strength at high inorganic nanoparticle content. Herein, cellulose nanofibers were extracted from wood and individually decorated with cobalt-ferrite nanoparticles and then for the first time molded at low temperature (ferrite and cellulose material. A nanocomposite with 70 wt % ferrite, 20 wt % cellulose nanofibers, and 10 wt % epoxy showed a modulus of 12.6 GPa, a tensile strength of 97 MPa, and a strain at failure of ca. 4%. Magnetic characterization was performed in a vibrating sample magnetometer, which showed that the coercivity was unaffected and that the saturation magnetization was in proportion with the ferrite content. The used ferrite, CoFe2O4, is a magnetically hard material, demonstrated by that the composite material behaved as a traditional permanent magnet. The presented processing route is easily adaptable to prepare millimeter-thick and moldable magnetic objects. This suggests that the processing method has the potential to be scaled-up for industrial use for the preparation of a new subcategory of magnetic, low-cost, and moldable objects based on cellulose nanofibers.

  13. The Gray Institute 'open' high-content, fluorescence lifetime microscopes.

    Science.gov (United States)

    Barber, P R; Tullis, I D C; Pierce, G P; Newman, R G; Prentice, J; Rowley, M I; Matthews, D R; Ameer-Beg, S M; Vojnovic, B

    2013-08-01

    We describe a microscopy design methodology and details of microscopes built to this 'open' design approach. These demonstrate the first implementation of time-domain fluorescence microscopy in a flexible automated platform with the ability to ease the transition of this and other advanced microscopy techniques from development to use in routine biology applications. This approach allows easy expansion and modification of the platform capabilities, as it moves away from the use of a commercial, monolithic, microscope body to small, commercial off-the-shelf and custom made modular components. Drawings and diagrams of our microscopes have been made available under an open license for noncommercial use at http://users.ox.ac.uk/~atdgroup. Several automated high-content fluorescence microscope implementations have been constructed with this design framework and optimized for specific applications with multiwell plates and tissue microarrays. In particular, three platforms incorporate time-domain FLIM via time-correlated single photon counting in an automated fashion. We also present data from experiments performed on these platforms highlighting their automated wide-field and laser scanning capabilities designed for high-content microscopy. Devices using these designs also form radiation-beam 'end-stations' at Oxford and Surrey Universities, showing the versatility and extendibility of this approach. © 2013 The Authors. Journal of Microscopy published by John Wiley & Sons Ltd on behalf of Royal Microscopical Society.

  14. Cytomegalovirus Destruction of Focal Adhesions Revealed in a High-Throughput Western Blot Analysis of Cellular Protein Expression† ▿

    OpenAIRE

    Stanton, Richard James; McSharry, Brian Patrick; Rickards, Carole Ruth; Wang, Edward Chung Yern; Tomasec, Peter; Wilkinson, Gavin William Grahame

    2007-01-01

    Human cytomegalovirus (HCMV) systematically manages the expression of cellular functions, rather than exerting the global shutoff of host cell protein synthesis commonly observed with other herpesviruses during the lytic cycle. While microarray technology has provided remarkable insights into viral control of the cellular transcriptome, HCMV is known to encode multiple mechanisms for posttranscriptional and posttranslation regulation of cellular gene expression. High-throughput Western blotti...

  15. Runaway electron dynamics in tokamak plasmas with high impurity content

    Science.gov (United States)

    Martín-Solís, J. R.; Loarte, A.; Lehnen, M.

    2015-09-01

    The dynamics of high energy runaway electrons is analyzed for plasmas with high impurity content. It is shown that modified collision terms are required in order to account for the collisions of the relativistic runaway electrons with partially stripped impurity ions, including the effect of the collisions with free and bound electrons, as well as the scattering by the full nuclear and the electron-shielded ion charge. The effect of the impurities on the avalanche runaway growth rate is discussed. The results are applied, for illustration, to the interpretation of the runaway electron behavior during disruptions, where large amounts of impurities are expected, particularly during disruption mitigation by massive gas injection. The consequences for the electron synchrotron radiation losses and the resulting runaway electron dynamics are also analyzed.

  16. High-content screening of functional genomic libraries.

    Science.gov (United States)

    Rines, Daniel R; Tu, Buu; Miraglia, Loren; Welch, Genevieve L; Zhang, Jia; Hull, Mitchell V; Orth, Anthony P; Chanda, Sumit K

    2006-01-01

    Recent advances in functional genomics have enabled genome-wide genetic studies in mammalian cells. These include the establishment of high-throughput transfection and viral propagation methodologies, the production of large-scale cDNA and siRNA libraries, and the development of sensitive assay detection processes and instrumentation. The latter has been significantly facilitated by the implementation of automated microscopy and quantitative image analysis, collectively referred to as high-content screening (HCS), toward cell-based functional genomics application. This technology can be applied to whole genome analysis of discrete molecular and phenotypic events at the level of individual cells and promises to significantly expand the scope of functional genomic analyses in mammalian cells. This chapter provides a comprehensive guide for curating and preparing function genomics libraries and performing HCS at the level of the genome.

  17. Multiplexed high-content analysis of mitochondrial morphofunction using live-cell microscopy.

    Science.gov (United States)

    Iannetti, Eligio F; Smeitink, Jan A M; Beyrath, Julien; Willems, Peter H G M; Koopman, Werner J H

    2016-09-01

    Mitochondria have a central role in cellular (patho)physiology, and they display a highly variable morphology that is probably coupled to their functional state. Here we present a protocol that allows unbiased and automated quantification of mitochondrial 'morphofunction' (i.e., morphology and membrane potential), cellular parameters (size, confluence) and nuclear parameters (number, morphology) in intact living primary human skin fibroblasts (PHSFs). Cells are cultured in 96-well plates and stained with tetramethyl rhodamine methyl ester (TMRM), calcein-AM (acetoxy-methyl ester) and Hoechst 33258. Next, multispectral fluorescence images are acquired using automated microscopy and processed to extract 44 descriptors. Subsequently, the descriptor data are subjected to a quality control (QC) algorithm based upon principal component analysis (PCA) and interpreted using univariate, bivariate and multivariate analysis. The protocol requires a time investment of ∼4 h distributed over 2 d. Although it is specifically developed for PHSFs, which are widely used in preclinical research, the protocol is portable to other cell types and can be scaled up for implementation in high-content screening.

  18. Dense and Cellular Zirconia Produced by Gel Casting with Agar: Preparation and High Temperature Characterization

    Directory of Open Access Journals (Sweden)

    Jean-Marc Tulliani

    2013-01-01

    Full Text Available A modified gel-casting process was developed to produce both dense and highly porous (40% volume yttria tetragonal zirconia polycrystal (Y-TZP using agar, a natural polysaccharide, as gelling agent. A fugitive phase, made of commercial polyethylene spheres, was added to the ceramic suspension before gelling to produce cellular ceramic structures. The characterization of the microstructural features of both dense and cellular ceramics was carried out by FEG SEM analysis of cross-sections produced by focused ion beam. The mechanical properties of the components were characterized at room temperature by nanoindentation tests in continuous stiffness measurement mode, by investigating the direct effect of the presence of residual microporosity. The presence of a diffuse residual microporosity from incomplete gel deaeration resulted in a decay of the bending strength and of the elastic modulus. The mechanical behavior of both dense and cellular zirconia (in terms of elastic modulus, flexural strength, and deformation at rupture was investigated by performing four-point bending tests at the temperature of 1500°C.

  19. High content screening for G protein-coupled receptors using cell-based protein translocation assays

    DEFF Research Database (Denmark)

    Grånäs, Charlotta; Lundholt, Betina Kerstin; Heydorn, Arne

    2005-01-01

    G protein-coupled receptors (GPCRs) have been one of the most productive classes of drug targets for several decades, and new technologies for GPCR-based discovery promise to keep this field active for years to come. While molecular screens for GPCR receptor agonist- and antagonist-based drugs...... as valuable discovery tools for several years. The application of high content cell-based screening to GPCR discovery has opened up additional possibilities, such as direct tracking of GPCRs, G proteins and other signaling pathway components using intracellular translocation assays. These assays provide...... the capability to probe GPCR function at the cellular level with better resolution than has previously been possible, and offer practical strategies for more definitive selectivity evaluation and counter-screening in the early stages of drug discovery. The potential of cell-based translocation assays for GPCR...

  20. The dust content of QSO hosts at high redshift

    CERN Document Server

    Calura, F; Vignali, C; Pozzi, F; Pipino, A; Matteucci, F

    2013-01-01

    Infrared observations of high-z quasar (QSO) hosts indicate the presence of large masses of dust in the early universe. When combined with other observables, such as neutral gas masses and star formation rates, the dust content of z~6 QSO hosts may help constraining their star formation history. We have collected a database of 58 sources from the literature discovered by various surveys and observed in the FIR. We have interpreted the available data by means of chemical evolution models for forming proto-spheroids, investigating the role of the major parameters regulating star formation and dust production. For a few systems, given the derived small dynamical masses, the observed dust content can be explained only assuming a top-heavy initial mass function, an enhanced star formation efficiency and an increased rate of dust accretion. However, the possibility that, for some systems, the dynamical mass has been underestimated cannot be excluded. If this were the case, the dust mass can be accounted for by stan...

  1. Automated analysis of high-content microscopy data with deep learning.

    Science.gov (United States)

    Kraus, Oren Z; Grys, Ben T; Ba, Jimmy; Chong, Yolanda; Frey, Brendan J; Boone, Charles; Andrews, Brenda J

    2017-04-18

    Existing computational pipelines for quantitative analysis of high-content microscopy data rely on traditional machine learning approaches that fail to accurately classify more than a single dataset without substantial tuning and training, requiring extensive analysis. Here, we demonstrate that the application of deep learning to biological image data can overcome the pitfalls associated with conventional machine learning classifiers. Using a deep convolutional neural network (DeepLoc) to analyze yeast cell images, we show improved performance over traditional approaches in the automated classification of protein subcellular localization. We also demonstrate the ability of DeepLoc to classify highly divergent image sets, including images of pheromone-arrested cells with abnormal cellular morphology, as well as images generated in different genetic backgrounds and in different laboratories. We offer an open-source implementation that enables updating DeepLoc on new microscopy datasets. This study highlights deep learning as an important tool for the expedited analysis of high-content microscopy data. © 2017 The Authors. Published under the terms of the CC BY 4.0 license.

  2. Detection of abnormally high amygdalin content in food by an enzyme immunoassay.

    Science.gov (United States)

    Cho, A-Yeon; Yi, Kye Sook; Rhim, Jung-Hyo; Kim, Kyu-Il; Park, Jae-Young; Keum, Eun-Hee; Chung, Junho; Oh, Sangsuk

    2006-04-30

    Amygdalin is a cyanogenic glycoside compound which is commonly found in the pits of many fruits and raw nuts. Although amygdalin itself is not toxic, it can release cyanide (CN) after hydrolysis when the pits and nuts are crushed, moistened and incubated, possibly within the gastrointestinal tract. CN reversibly inhibits cellular oxidizing enzymes and cyanide poisoning generates a range of clinical symptoms. As some pits and nuts may contain unusually high levels of amygdalin such that there is a sufficient amount to induce critical CN poisoning in humans, the detection of abnormal content of amygdalin in those pits and nuts can be a life-saving measure. Although there are various methods to detect amygdalin in food extracts, an enzyme immunoassay has not been developed for this purpose. In this study we immunized New Zealand White rabbits with an amygdalin-KLH (keyhole limpet hemocyanin) conjugate and succeeded in raising anti-sera reactive to amygdalin, proving that amygdalin can behave as a hapten in rabbits. Using this polyclonal antibody, we developed a competition enzyme immunoassay for determination of amygdalin concentration in aqueous solutions. This technique was able to effectively detect abnormally high amygdalin content in various seeds and nuts. In conclusion, we proved that enzyme immunoassay can be used to determine the amount of amygdalin in food extracts, which will allow automated analysis with high throughput.

  3. HIGH TEMPERATURE EFFECT ON TOMATO (Lycopersicon esculentum PIGMENT AND PROTEIN CONTENT AND CELLULAR VIABILITY

    Directory of Open Access Journals (Sweden)

    Daymi Camejo

    2001-01-01

    Full Text Available Cultivares de tomate con diferente sensibilidad a las altas temperaturas (Campbell-28 y Amalia y el tipo silvestre Nagcarlang, fueron expuestos a diferentes condiciones de estrés (45oC-dos horas de exposición, 45oC-tres horas de exposición y 25oC como control. Los contenidos de clorofila “a”, “b” y total fueron determinados, así como los de carotenoides y proteinas totales solubles. La relación clorofila “a”/“b” y clorofila/carotenoides fue estimada a partir de los resultados obtenidos. Las variables evaluadas fueron modificadas según el cultivar y la condición de estrés. La selección de Nagcarlang como modelo de termotolerancia permite establecer que variables tales como: relación de clorofila “a”/“b”, clorofila/carotenoides, contenido de proteínas totales solubles y viabilidad celular, pudieran ser útiles para describir cultivares tolerantes a las altas temperaturas, dependiendo del tiempo de exposición al estrés.

  4. Semisolid Slurry Preparation of Die Steel with High Chromium Content

    Institute of Scientific and Technical Information of China (English)

    MAO Wei-min; ZHAO Ai-min; ZHANG Li-juan; ZHONG Xue-you

    2004-01-01

    The semisolid slurry preparation of die steels Cr12 and Cr12MoV with high chromium content was studied. The results show that the semisolid slurry of both steels with solid of 40 %-60 % can be made by electromagnetic stirring method and is easy to be discharged from the bottom little hole of the stirring chamber. The sizes of the spherical primary austenite in the slurry of die steels Cr12 and Cr12MoV are 50-100 μm and 80-150 μm, respectively. The homogeneous temperature field and solute field for both steel melts are obtained. The strong temperature fluctuation in the melt with many fine primary austenite grains occurs and the remelting of the secondary arm roots at the same time is accelerated because of the electromagnetic stirring. These are the most important reasons for deposition of spherical primary austenite grains.

  5. High-Content Screening for Quantitative Cell Biology.

    Science.gov (United States)

    Mattiazzi Usaj, Mojca; Styles, Erin B; Verster, Adrian J; Friesen, Helena; Boone, Charles; Andrews, Brenda J

    2016-08-01

    High-content screening (HCS), which combines automated fluorescence microscopy with quantitative image analysis, allows the acquisition of unbiased multiparametric data at the single cell level. This approach has been used to address diverse biological questions and identify a plethora of quantitative phenotypes of varying complexity in numerous different model systems. Here, we describe some recent applications of HCS, ranging from the identification of genes required for specific biological processes to the characterization of genetic interactions. We review the steps involved in the design of useful biological assays and automated image analysis, and describe major challenges associated with each. Additionally, we highlight emerging technologies and future challenges, and discuss how the field of HCS might be enhanced in the future.

  6. Preparation of Garlic Powder with High Allicin Content

    Institute of Scientific and Technical Information of China (English)

    LI Yu; XU Shi-ying

    2007-01-01

    Garlic powder with high allicin content was prepared using microwave-vacuum and vacuum drying as well as microencapsulation to protect alliinase activity throughout the stomach and improve the ratio of alliin transforming into allicin. The results showed that the optimal drying condition was 376.1 W for 3 min, 282.1 W for 3 min, 188 W for 9 min, and 94 W for 3 min. The thiosulfinates retention after drying was 90.2%. Following drying, the garlic powder was microencapsulated by modified fluidized bed technique. Scanning electron microscope revealed good integrity and core materials that were embedded in the microcapsules. Studies on the release kinetics of microencapsulated garlic granulates in vitro using simulated intestinal fluid indicated that release of garlic powder could be controlled in the intestine by passing stomach conditions.

  7. Oxy-combustion of high water content fuels

    Science.gov (United States)

    Yi, Fei

    As the issues of global warming and the energy crisis arouse extensive concern, more and more research is focused on maximizing energy efficiency and capturing CO2 in power generation. To achieve this, in this research, we propose an unconventional concept of combustion - direct combustion of high water content fuels. Due to the high water content in the fuels, they may not burn under air-fired conditions. Therefore, oxy-combustion is applied. Three applications of this concept in power generation are proposed - direct steam generation for the turbine cycle, staged oxy-combustion with zero flue gas recycle, and oxy-combustion in a low speed diesel-type engine. The proposed processes could provide alternative approaches to directly utilize fuels which intrinsically have high water content. A large amount of energy to remove the water, when the fuels are utilized in a conventional approach, is saved. The properties and difficulty in dewatering high water content fuels (e.g. bioethanol, microalgae and fine coal) are summarized. These fuels include both renewable and fossil fuels. In addition, the technique can also allow for low-cost carbon capture due to oxy-combustion. When renewable fuel is utilized, the whole process can be carbon negative. To validate and evaluate this concept, the research focused on the investigation of the flame stability and characteristics for high water content fuels. My study has demonstrated the feasibility of burning fuels that have been heavily diluted with water in a swirl-stabilized burner. Ethanol and 1-propanol were first tested as the fuels and the flame stability maps were obtained. Flame stability, as characterized by the blow-off limit -- the lowest O2 concentration when a flame could exist under a given oxidizer flow rate, was determined as a function of total oxidizer flow rate, fuel concentration and nozzle type. Furthermore, both the gas temperature contour and the overall ethanol concentration in the droplets along the

  8. High content analysis of phagocytic activity and cell morphology with PuntoMorph.

    Science.gov (United States)

    Al-Ali, Hassan; Gao, Han; Dalby-Hansen, Camilla; Peters, Vanessa Ann; Shi, Yan; Brambilla, Roberta

    2017-11-01

    Phagocytosis is essential for maintenance of normal homeostasis and healthy tissue. As such, it is a therapeutic target for a wide range of clinical applications. The development of phenotypic screens targeting phagocytosis has lagged behind, however, due to the difficulties associated with image-based quantification of phagocytic activity. We present a robust algorithm and cell-based assay system for high content analysis of phagocytic activity. The method utilizes fluorescently labeled beads as a phagocytic substrate with defined physical properties. The algorithm employs statistical modeling to determine the mean fluorescence of individual beads within each image, and uses the information to conduct an accurate count of phagocytosed beads. In addition, the algorithm conducts detailed and sophisticated analysis of cellular morphology, making it a standalone tool for high content screening. We tested our assay system using microglial cultures. Our results recapitulated previous findings on the effects of microglial stimulation on cell morphology and phagocytic activity. Moreover, our cell-level analysis revealed that the two phenotypes associated with microglial activation, specifically cell body hypertrophy and increased phagocytic activity, are not highly correlated. This novel finding suggests the two phenotypes may be under the control of distinct signaling pathways. We demonstrate that our assay system outperforms preexisting methods for quantifying phagocytic activity in multiple dimensions including speed, accuracy, and resolution. We provide a framework to facilitate the development of high content assays suitable for drug screening. For convenience, we implemented our algorithm in a standalone software package, PuntoMorph. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. CONTENTS

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    The Development and Evolution of the Idea of the Mandate of Heaven in the Zhou Dynasty The changes in the idea of Mandate of Heaven during the Shang and Zhou dynasties are of great significance in the course of the development of traditional Chinese culture. The quickening and awakening of the humanistic spirit was not the entire content of the Zhou idea of Mandate of Heaven. In the process of annihilating the Shang dynasty and setting up their state, the Zhou propagated the idea of the Mandate of Heaven out of practical needs. Their idea of the Mandate of Heaven was not very different from that of the Shang. From the Western Zhou on, the Zhou idea of Mandate of Heaven by no means developed in a linear way along a rational track. The intermingling of rationality and irrationality and of awakening and non-awakening remained the overall state of the Zhou intellectual superstructure after their "spiritual awakening".

  10. NeuriteQuant: An open source toolkit for high content screens of neuronal Morphogenesis

    Directory of Open Access Journals (Sweden)

    Hwang Eric

    2011-10-01

    Full Text Available Abstract Background To date, some of the most useful and physiologically relevant neuronal cell culture systems, such as high density co-cultures of astrocytes and primary hippocampal neurons, or differentiated stem cell-derived cultures, are characterized by high cell density and partially overlapping cellular structures. Efficient analytical strategies are required to enable rapid, reliable, quantitative analysis of neuronal morphology in these valuable model systems. Results Here we present the development and validation of a novel bioinformatics pipeline called NeuriteQuant. This tool enables fully automated morphological analysis of large-scale image data from neuronal cultures or brain sections that display a high degree of complexity and overlap of neuronal outgrowths. It also provides an efficient web-based tool to review and evaluate the analysis process. In addition to its built-in functionality, NeuriteQuant can be readily extended based on the rich toolset offered by ImageJ and its associated community of developers. As proof of concept we performed automated screens for modulators of neuronal development in cultures of primary neurons and neuronally differentiated P19 stem cells, which demonstrated specific dose-dependent effects on neuronal morphology. Conclusions NeuriteQuant is a freely available open-source tool for the automated analysis and effective review of large-scale high-content screens. It is especially well suited to quantify the effect of experimental manipulations on physiologically relevant neuronal cultures or brain sections that display a high degree of complexity and overlap among neurites or other cellular structures.

  11. Punicalagin exerts protective effect against high glucose-induced cellular stress and neural tube defects.

    Science.gov (United States)

    Zhong, Jianxiang; Reece, E Albert; Yang, Peixin

    2015-11-13

    Maternal diabetes-induced birth defects remain a significant health problem. Studying the effect of natural compounds with antioxidant properties and minimal toxicities on diabetic embryopathy may lead to the development of new and safe dietary supplements. Punicalagin is a primary polyphenol found in pomegranate juice, which possesses antioxidant, anti-inflammatory and anti-tumorigenic properties, suggesting a protective effect of punicalagin on diabetic embryopathy. Here, we examined whether punicalagin could reduce high glucose-induced neural tube defects (NTDs), and if this rescue occurs through blockage of cellular stress and caspase activation. Embryonic day 8.5 (E8.5) mouse embryos were cultured for 24 or 36 h with normal (5 mM) glucose or high glucose (16.7 mM), in presence or absence of 10 or 20 μM punicalagin. 10 μM punicalagin slightly reduced NTD formation under high glucose conditions; however, 20 μM punicalagin significantly inhibited high glucose-induced NTD formation. Punicalagin suppressed high glucose-induced lipid peroxidation marker 4-hydroxynonenal, nitrotyrosine-modified proteins, and lipid peroxides. Moreover, punicalagin abrogated endoplasmic reticulum stress by inhibiting phosphorylated protein kinase ribonucleic acid (RNA)-like ER kinase (p-PERK), phosphorylated inositol-requiring protein-1α (p-IRE1α), phosphorylated eukaryotic initiation factor 2α (p-eIF2α), C/EBP-homologous protein (CHOP), binding immunoglobulin protein (BiP) and x-box binding protein 1 (XBP1) mRNA splicing. Additionally, punicalagin suppressed high glucose-induced caspase 3 and caspase 8 cleavage. Punicalagin reduces high glucose-induced NTD formation by blocking cellular stress and caspase activation. These observations suggest punicalagin supplements could mitigate the teratogenic effects of hyperglycemia in the developing embryo, and possibly prevent diabetes-induced NTDs.

  12. Optical High Content Nanoscopy of Epigenetic Marks Decodes Phenotypic Divergence in Stem Cells

    Science.gov (United States)

    Kim, Joseph J.; Bennett, Neal K.; Devita, Mitchel S.; Chahar, Sanjay; Viswanath, Satish; Lee, Eunjee A.; Jung, Giyoung; Shao, Paul P.; Childers, Erin P.; Liu, Shichong; Kulesa, Anthony; Garcia, Benjamin A.; Becker, Matthew L.; Hwang, Nathaniel S.; Madabhushi, Anant; Verzi, Michael P.; Moghe, Prabhas V.

    2017-01-01

    While distinct stem cell phenotypes follow global changes in chromatin marks, single-cell chromatin technologies are unable to resolve or predict stem cell fates. We propose the first such use of optical high content nanoscopy of histone epigenetic marks (epi-marks) in stem cells to classify emergent cell states. By combining nanoscopy with epi-mark textural image informatics, we developed a novel approach, termed EDICTS (Epi-mark Descriptor Imaging of Cell Transitional States), to discern chromatin organizational changes, demarcate lineage gradations across a range of stem cell types and robustly track lineage restriction kinetics. We demonstrate the utility of EDICTS by predicting the lineage progression of stem cells cultured on biomaterial substrates with graded nanotopographies and mechanical stiffness, thus parsing the role of specific biophysical cues as sensitive epigenetic drivers. We also demonstrate the unique power of EDICTS to resolve cellular states based on epi-marks that cannot be detected via mass spectrometry based methods for quantifying the abundance of histone post-translational modifications. Overall, EDICTS represents a powerful new methodology to predict single cell lineage decisions by integrating high content super-resolution nanoscopy and imaging informatics of the nuclear organization of epi-marks. PMID:28051095

  13. TGF-β1-mediated differentiation of fibroblasts is associated with increased mitochondrial content and cellular respiration.

    Science.gov (United States)

    Negmadjanov, Ulugbek; Godic, Zarko; Rizvi, Farhan; Emelyanova, Larisa; Ross, Gracious; Richards, John; Holmuhamedov, Ekhson L; Jahangir, Arshad

    2015-01-01

    Cytokine-dependent activation of fibroblasts to myofibroblasts, a key event in fibrosis, is accompanied by phenotypic changes with increased secretory and contractile properties dependent on increased energy utilization, yet changes in the energetic profile of these cells are not fully described. We hypothesize that the TGF-β1-mediated transformation of myofibroblasts is associated with an increase in mitochondrial content and function when compared to naive fibroblasts. Cultured NIH/3T3 mouse fibroblasts treated with TGF-β1, a profibrotic cytokine, or vehicle were assessed for transformation to myofibroblasts (appearance of α-smooth muscle actin [α-SMA] stress fibers) and associated changes in mitochondrial content and functions using laser confocal microscopy, Seahorse respirometry, multi-well plate reader and biochemical protocols. Expression of mitochondrial-specific proteins was determined using western blotting, and the mitochondrial DNA quantified using Mitochondrial DNA isolation kit. Treatment with TGF-β1 (5 ng/mL) induced transformation of naive fibroblasts into myofibroblasts with a threefold increase in the expression of α-SMA (6.85 ± 0.27 RU) compared to cells not treated with TGF-β1 (2.52 ± 0.11 RU). TGF-β1 exposure increased the number of mitochondria in the cells, as monitored by membrane potential sensitive dye tetramethylrhodamine, and expression of mitochondria-specific proteins; voltage-dependent anion channels (0.54 ± 0.05 vs. 0.23 ± 0.05 RU) and adenine nucleotide transporter (0.61 ± 0.11 vs. 0.22 ± 0.05 RU), as well as mitochondrial DNA content (530 ± 12 μg DNA/106 cells vs. 307 ± 9 μg DNA/106 cells in control). TGF-β1 treatment was associated with an increase in mitochondrial function with a twofold increase in baseline oxygen consumption rate (2.25 ± 0.03 vs. 1.13 ± 0.1 nmol O2/min/106 cells) and FCCP-induced mitochondrial respiration (2.87 ± 0.03 vs. 1.46 ± 0.15 nmol O2/min/106 cells). TGF-β1 induced differentiation

  14. TGF-β1-mediated differentiation of fibroblasts is associated with increased mitochondrial content and cellular respiration.

    Directory of Open Access Journals (Sweden)

    Ulugbek Negmadjanov

    Full Text Available Cytokine-dependent activation of fibroblasts to myofibroblasts, a key event in fibrosis, is accompanied by phenotypic changes with increased secretory and contractile properties dependent on increased energy utilization, yet changes in the energetic profile of these cells are not fully described. We hypothesize that the TGF-β1-mediated transformation of myofibroblasts is associated with an increase in mitochondrial content and function when compared to naive fibroblasts.Cultured NIH/3T3 mouse fibroblasts treated with TGF-β1, a profibrotic cytokine, or vehicle were assessed for transformation to myofibroblasts (appearance of α-smooth muscle actin [α-SMA] stress fibers and associated changes in mitochondrial content and functions using laser confocal microscopy, Seahorse respirometry, multi-well plate reader and biochemical protocols. Expression of mitochondrial-specific proteins was determined using western blotting, and the mitochondrial DNA quantified using Mitochondrial DNA isolation kit.Treatment with TGF-β1 (5 ng/mL induced transformation of naive fibroblasts into myofibroblasts with a threefold increase in the expression of α-SMA (6.85 ± 0.27 RU compared to cells not treated with TGF-β1 (2.52 ± 0.11 RU. TGF-β1 exposure increased the number of mitochondria in the cells, as monitored by membrane potential sensitive dye tetramethylrhodamine, and expression of mitochondria-specific proteins; voltage-dependent anion channels (0.54 ± 0.05 vs. 0.23 ± 0.05 RU and adenine nucleotide transporter (0.61 ± 0.11 vs. 0.22 ± 0.05 RU, as well as mitochondrial DNA content (530 ± 12 μg DNA/106 cells vs. 307 ± 9 μg DNA/106 cells in control. TGF-β1 treatment was associated with an increase in mitochondrial function with a twofold increase in baseline oxygen consumption rate (2.25 ± 0.03 vs. 1.13 ± 0.1 nmol O2/min/106 cells and FCCP-induced mitochondrial respiration (2.87 ± 0.03 vs. 1.46 ± 0.15 nmol O2/min/106 cells.TGF-β1 induced

  15. High content cell-based assay for the inflammatory pathway

    Science.gov (United States)

    Mukherjee, Abhishek; Song, Joon Myong

    2015-07-01

    Cellular inflammation is a non-specific immune response to tissue injury that takes place via cytokine network orchestration to maintain normal tissue homeostasis. However chronic inflammation that lasts for a longer period, plays the key role in human diseases like neurodegenerative disorders and cancer development. Understanding the cellular and molecular mechanisms underlying the inflammatory pathways may be effective in targeting and modulating their outcome. Tumor necrosis factor alpha (TNF-α) is a pro-inflammatory cytokine that effectively combines the pro-inflammatory features with the pro-apoptotic potential. Increased levels of TNF-α observed during acute and chronic inflammatory conditions are believed to induce adverse phenotypes like glucose intolerance and abnormal lipid profile. Natural products e. g., amygdalin, cinnamic acid, jasmonic acid and aspirin have proven efficacy in minimizing the TNF-α induced inflammation in vitro and in vivo. Cell lysis-free quantum dot (QDot) imaging is an emerging technique to identify the cellular mediators of a signaling cascade with a single assay in one run. In comparison to organic fluorophores, the inorganic QDots are bright, resistant to photobleaching and possess tunable optical properties that make them suitable for long term and multicolor imaging of various components in a cellular crosstalk. Hence we tested some components of the mitogen activated protein kinase (MAPK) pathway during TNF-α induced inflammation and the effects of aspirin in HepG2 cells by QDot multicolor imaging technique. Results demonstrated that aspirin showed significant protective effects against TNF-α induced cellular inflammation. The developed cell based assay paves the platform for the analysis of cellular components in a smooth and reliable way.

  16. Aldolase B knockdown prevents high glucose-induced methylglyoxal overproduction and cellular dysfunction in endothelial cells.

    Directory of Open Access Journals (Sweden)

    Jianghai Liu

    Full Text Available We used cultured endothelial cells as a model to examine whether up-regulation of aldolase B and enhanced methylglyoxal (MG formation play an important role in high glucose-induced overproduction of advanced glycosylation endproducts (AGEs, oxidative stress and cellular dysfunction. High glucose (25 mM incubation up-regulated mRNA levels of aldose reductase (an enzyme converting glucose to fructose and aldolase B (a key enzyme that catalyzes MG formation from fructose and enhanced MG formation in human umbilical vein endothelial cells (HUVECs and HUVEC-derived EA. hy926 cells. High glucose-increased MG production in EA. hy926 cells was completely prevented by siRNA knockdown of aldolase B, but unaffected by siRNA knockdown of aldolase A, an enzyme responsible for MG formation during glycolysis. In addition, inhibition of cytochrome P450 2E1 or semicarbazide-sensitive amine oxidase which produces MG during the metabolism of lipid and proteins, respectively, did not alter MG production. Both high glucose (25 mM and MG (30, 100 µM increased the formation of N(ε-carboxyethyl-lysine (CEL, a MG-induced AGE, oxidative stress (determined by the generation of oxidized DCF, H(2O(2, protein carbonyls and 8-oxo-dG, O-GlcNAc modification (product of the hexosamine pathway, membrane protein kinase C activity and nuclear translocation of NF-κB in EA. hy926 cells. However, the above metabolic and signaling alterations induced by high glucose were completely prevented by knockdown of aldolase B and partially by application of aminoguanidine (a MG scavenger or alagebrium (an AGEs breaker. In conclusion, efficient inhibition of aldolase B can prevent high glucose-induced overproduction of MG and related cellular dysfunction in endothelial cells.

  17. An oral multispecies biofilm model for high content screening applications

    Science.gov (United States)

    Kommerein, Nadine; Stumpp, Sascha N.; Müsken, Mathias; Ehlert, Nina; Winkel, Andreas; Häussler, Susanne; Behrens, Peter; Buettner, Falk F. R.; Stiesch, Meike

    2017-01-01

    Peri-implantitis caused by multispecies biofilms is a major complication in dental implant treatment. The bacterial infection surrounding dental implants can lead to bone loss and, in turn, to implant failure. A promising strategy to prevent these common complications is the development of implant surfaces that inhibit biofilm development. A reproducible and easy-to-use biofilm model as a test system for large scale screening of new implant surfaces with putative antibacterial potency is therefore of major importance. In the present study, we developed a highly reproducible in vitro four-species biofilm model consisting of the highly relevant oral bacterial species Streptococcus oralis, Actinomyces naeslundii, Veillonella dispar and Porphyromonas gingivalis. The application of live/dead staining, quantitative real time PCR (qRT-PCR), scanning electron microscopy (SEM) and urea-NaCl fluorescence in situ hybridization (urea-NaCl-FISH) revealed that the four-species biofilm community is robust in terms of biovolume, live/dead distribution and individual species distribution over time. The biofilm community is dominated by S. oralis, followed by V. dispar, A. naeslundii and P. gingivalis. The percentage distribution in this model closely reflects the situation in early native plaques and is therefore well suited as an in vitro model test system. Furthermore, despite its nearly native composition, the multispecies model does not depend on nutrient additives, such as native human saliva or serum, and is an inexpensive, easy to handle and highly reproducible alternative to the available model systems. The 96-well plate format enables high content screening for optimized implant surfaces impeding biofilm formation or the testing of multiple antimicrobial treatment strategies to fight multispecies biofilm infections, both exemplary proven in the manuscript. PMID:28296966

  18. Cellular mechanisms of reduced sarcoplasmic reticulum Ca2+ content in L-thyroxin-induced rat ventricular hypertrophy

    Institute of Scientific and Technical Information of China (English)

    Lai-jing SONG; Guan-lei WANG; Jie LIU; Qin-ying QIU; Jing-hua OU; Yong-yuan GUAN

    2008-01-01

    Aim:To examine how the sarcoplasmic reticulum (SR) Ca2+ content changes and the underlying mechanism in L-thyroxin-induced cardiac hypertrophy. Methods:Echocardiography was used to confirm the establishment of the cardiac hypertro-phy model. The confocal microscopy and fluorescent indicator Fluo-3 was ap-plied to examine the intracellular Ca2+ concentration ([Ca2+]I), the Ca2+ sparks, and the caffeine-induced Ca2+ transient in freshly isolated cardiac ventricular myocytes. The activity of sarcolemmal and SR Ca2+-ATPase 2a (SERCA2a) in the ventricular tissue was also measured, respectively. Results:L-thyroxin (1 mg/kg injection for 10 d) induces left ventricular cardiac hypertrophy with normal myocardial function. The decreased caffeine-induced Ca2+ transient in the Ca2+-free solution was detected. The spontaneous Ca2+ sparks in hypertrophied myocytes occurred more frequently than in normal cells, with similar duration and spatial spread, but smaller amplitude. Then the basal [Ca2+]I increase was observed in quiescent left ventricular myocytes from hyperthyroidism rats. The activity of sarcolemmal and SR Ca2+-ATPase was decreased in the hypertrophied ventricle tissue. Conclusion:The results suggested that the reduced SR Ca2+ content may be associated with an increased Ca2+ leak and reduced SERCA2a activity, contributing to abnormal intracellular Ca2+ handling during hypertrophy in hyperthyroidism rats.

  19. Highly efficient enzyme encapsulation in a protein nanocage: towards enzyme catalysis in a cellular nanocompartment mimic

    Science.gov (United States)

    Schoonen, Lise; Nolte, Roeland J. M.; van Hest, Jan C. M.

    2016-07-01

    The study of enzyme behavior in small nanocompartments is crucial for the understanding of biocatalytic processes in the cellular environment. We have developed an enzymatic conjugation strategy to attach a model enzyme to the interior of a cowpea chlorotic mottle virus capsid. It is shown that with this methodology high encapsulation efficiencies can be achieved. Additionally, we demonstrate that the encapsulation does not affect the enzyme performance in terms of a decreased activity or a hampered substrate diffusion. Finally, it is shown that the encapsulated enzymes are protected against proteases. We believe that our strategy can be used to study enzyme kinetics in an environment that approaches physiological conditions.The study of enzyme behavior in small nanocompartments is crucial for the understanding of biocatalytic processes in the cellular environment. We have developed an enzymatic conjugation strategy to attach a model enzyme to the interior of a cowpea chlorotic mottle virus capsid. It is shown that with this methodology high encapsulation efficiencies can be achieved. Additionally, we demonstrate that the encapsulation does not affect the enzyme performance in terms of a decreased activity or a hampered substrate diffusion. Finally, it is shown that the encapsulated enzymes are protected against proteases. We believe that our strategy can be used to study enzyme kinetics in an environment that approaches physiological conditions. Electronic supplementary information (ESI) available: Experimental procedures for the cloning, expression, and purification of all proteins, as well as supplementary figures and calculations. See DOI: 10.1039/c6nr04181g

  20. Fluorescence Modified Chitosan-Coated Magnetic Nanoparticles for High-Efficient Cellular Imaging

    Directory of Open Access Journals (Sweden)

    Nie Fang

    2009-01-01

    Full Text Available Abstract Labeling of cells with nanoparticles for living detection is of interest to various biomedical applications. In this study, novel fluorescent/magnetic nanoparticles were prepared and used in high-efficient cellular imaging. The nanoparticles coated with the modified chitosan possessed a magnetic oxide core and a covalently attached fluorescent dye. We evaluated the feasibility and efficiency in labeling cancer cells (SMMC-7721 with the nanoparticles. The nanoparticles exhibited a high affinity to cells, which was demonstrated by flow cytometry and magnetic resonance imaging. The results showed that cell-labeling efficiency of the nanoparticles was dependent on the incubation time and nanoparticles’ concentration. The minimum detected number of labeled cells was around 104by using a clinical 1.5-T MRI imager. Fluorescence and transmission electron microscopy instruments were used to monitor the localization patterns of the magnetic nanoparticles in cells. These new magneto-fluorescent nanoagents have demonstrated the potential for future medical use.

  1. Label-free imaging of cellular malformation using high resolution photoacoustic microscopy

    Science.gov (United States)

    Chen, Zhongjiang; Li, Bingbing; Yang, Sihua

    2014-09-01

    A label-free high resolution photoacoustic microscopy (PAM) system for imaging cellular malformation is presented. The carbon fibers were used to testify the lateral resolution of the PAM. Currently, the lateral resolution is better than 2.7 μm. The human normal red blood cells (RBCs) were used to prove the imaging capability of the system, and a single red blood cell was mapped with high contrast. Moreover, the iron deficiency anemia RBCs were clearly distinguished from the cell morphology by using the PAM. The experimental results demonstrate that the photoacoustic microscopy system can accomplish label-free photoacoustic imaging and that it has clinical potential for use in the detection of erythrocytes and blood vessels malformation.

  2. Downregulation of cellular protective factors of rumen epithelium in goats fed high energy diet.

    Directory of Open Access Journals (Sweden)

    Manfred Hollmann

    Full Text Available Energy-rich diets can challenge metabolic and protective functions of the rumen epithelial cells, but the underlying factors are unclear. This study sought to evaluate proteomic changes of the rumen epithelium in goats fed a low, medium, or high energy diet. Expression of protein changes were compared by two-dimensional differential gel electrophoresis followed by protein identification with matrix assisted laser desorption ionisation tandem time-of-flight mass spectrometry. Of about 2,000 spots commonly detected in all gels, 64 spots were significantly regulated, which were traced back to 24 unique proteins. Interestingly, the expression profiles of several chaperone proteins with important cellular protective functions such as heat shock cognate 71 kDa protein, peroxiredoxin-6, serpin H1, protein disulfide-isomerase, and selenium-binding protein were collectively downregulated in response to high dietary energy supply. Similar regulation patterns were obtained for some other proteins involved in transport or metabolic functions. In contrast, metabolic enzymes like retinal dehydrogenase 1 and ATP synthase subunit beta, mitochondrial precursor were upregulated in response to high energy diet. Lower expressions of chaperone proteins in the rumen epithelial cells in response to high energy supply may suggest that these cells were less protected against the potentially harmful rumen toxic compounds, which might have consequences for rumen and systemic health. Our findings also suggest that energy-rich diets and the resulting acidotic insult may render rumen epithelial cells more vulnerable to cellular damage by attenuating their cell defense system, hence facilitating the impairment of rumen barrier function, typically observed in energy-rich fed ruminants.

  3. Downregulation of Cellular Protective Factors of Rumen Epithelium in Goats Fed High Energy Diet

    Science.gov (United States)

    Hollmann, Manfred; Miller, Ingrid; Hummel, Karin; Sabitzer, Sonja; Metzler-Zebeli, Barbara U.; Razzazi-Fazeli, Ebrahim; Zebeli, Qendrim

    2013-01-01

    Energy-rich diets can challenge metabolic and protective functions of the rumen epithelial cells, but the underlying factors are unclear. This study sought to evaluate proteomic changes of the rumen epithelium in goats fed a low, medium, or high energy diet. Expression of protein changes were compared by two-dimensional differential gel electrophoresis followed by protein identification with matrix assisted laser desorption ionisation tandem time-of-flight mass spectrometry. Of about 2,000 spots commonly detected in all gels, 64 spots were significantly regulated, which were traced back to 24 unique proteins. Interestingly, the expression profiles of several chaperone proteins with important cellular protective functions such as heat shock cognate 71 kDa protein, peroxiredoxin-6, serpin H1, protein disulfide-isomerase, and selenium-binding protein were collectively downregulated in response to high dietary energy supply. Similar regulation patterns were obtained for some other proteins involved in transport or metabolic functions. In contrast, metabolic enzymes like retinal dehydrogenase 1 and ATP synthase subunit beta, mitochondrial precursor were upregulated in response to high energy diet. Lower expressions of chaperone proteins in the rumen epithelial cells in response to high energy supply may suggest that these cells were less protected against the potentially harmful rumen toxic compounds, which might have consequences for rumen and systemic health. Our findings also suggest that energy-rich diets and the resulting acidotic insult may render rumen epithelial cells more vulnerable to cellular damage by attenuating their cell defense system, hence facilitating the impairment of rumen barrier function, typically observed in energy-rich fed ruminants. PMID:24349094

  4. Synthetic High-Density Lipoprotein-Like Nanocarrier Improved Cellular Transport of Lysosomal Cholesterol in Human Sterol Carrier Protein-Deficient Fibroblasts.

    Science.gov (United States)

    Nam, Da-Eun; Kim, Ok-Kyung; Park, Yoo Kyoung; Lee, Jeongmin

    2016-01-01

    Sterol carrier protein-2 (SCP-2), which is not found in tissues of people with Zellweger syndrome, facilitates the movement of cholesterol within cells, resulting in abnormal accumulation of cholesterol in SCP-2-deficient cells. This study investigated whether synthetic high-density lipoprotein-like nanocarrier (sHDL-NC) improves the cellular transport of lysosomal cholesterol to plasma membrane in SCP-2-deficient fibroblasts. Human SCP-2-deficient fibroblasts were incubated with [(3)H-cholesterol]LDL as a source of cholesterol and sHDL-NC. The cells were fractionated by centrifugation permit tracking of [(3)H]-cholesterol from lysosome into plasma membrane. Furthermore, cellular content of cholesteryl ester as a storage form and mRNA expression of low-density lipoprotein (LDL) receptor were measured to support the cholesterol transport to plasma membrane. Incubation with sHDL-NC for 8 h significantly increased uptake of [(3)H]-cholesterol to lysosome by 53% and further enhanced the transport of [(3)H]-cholesterol to plasma membrane by 32%. Treatment with sHDL-NC significantly reduced cellular content of cholesteryl ester and increased mRNA expression of LDL receptor (LDL-R). In conclusion, sHDL-NC enables increased transport of lysosomal cholesterol to plasma membrane. In addition, these data were indirectly supported by decreased cellular content of cholesteryl ester and increased gene expression of LDL-R. Therefore, sHDL-NC may be a useful vehicle for transporting cholesterol, which may help to prevent accumulation of cholesterol in SCP-2-deficient fibroblasts.

  5. [Nicotinic acid increases cellular transport of high density lipoprotein cholesterol in patients with hypoalphalipoproteinemia].

    Science.gov (United States)

    Figueroa, Catalina; Droppelmann, Katherine; Quiñones, Verónica; Amigo, Ludwig; Mendoza, Camila; Serrano, Valentina; Véjar, Margarita; Maiz, Alberto; Rigotti, Attilio

    2015-09-01

    Plasma high density lipoproteins (HDL) are involved in reverse cholesterol transport mediated by the scavenger receptor class B type I (SR-BI). Nicotinic acid increases HDL cholesterol levels, even though its specific impact on SR-BI dependent-cellular cholesterol transport remains unknown. To determine the effect of nicotinic acid on HDL particle functionality in cholesterol efflux and uptake mediated by SR-BI in cultured cells in hypoalphalipoproteinemic patients. In a pilot study, eight patients with low HDL (≤ 40 mg/dL) were treated with extended release nicotinic acid. HDL cholesterol and phospholipid levels, HDL2 and HDL3 fractions and HDL particle sizes were measured at baseline and post-therapy. Before and after nicotinic acid treatment, HDL particles were used for cholesterol transport studies in cells transfected with SR-BI. Nicotinic acid treatment raised total HDL cholesterol and phospholipids, HDL2 levels as well as HDL particle size. Nicotinic acid significantly increased HDL cholesterol efflux and uptake capacity mediated by SR-BI in cultured cells. Nicotinic acid therapy increases SR-BI-dependent HDL cholesterol transport in cultured cells, establishing a new cellular mechanism by which this lipid-lowering drug appears to modulate HDL metabolism in patients with hypoalphalipoproteinemia.

  6. High-content, high-throughput screening for the identification of cytotoxic compounds based on cell morphology and cell proliferation markers.

    Directory of Open Access Journals (Sweden)

    Heather L Martin

    Full Text Available Toxicity is a major cause of failure in drug discovery and development, and whilst robust toxicological testing occurs, efficiency could be improved if compounds with cytotoxic characteristics were identified during primary compound screening. The use of high-content imaging in primary screening is becoming more widespread, and by utilising phenotypic approaches it should be possible to incorporate cytotoxicity counter-screens into primary screens. Here we present a novel phenotypic assay that can be used as a counter-screen to identify compounds with adverse cellular effects. This assay has been developed using U2OS cells, the PerkinElmer Operetta high-content/high-throughput imaging system and Columbus image analysis software. In Columbus, algorithms were devised to identify changes in nuclear morphology, cell shape and proliferation using DAPI, TOTO-3 and phosphohistone H3 staining, respectively. The algorithms were developed and tested on cells treated with doxorubicin, taxol and nocodazole. The assay was then used to screen a novel, chemical library, rich in natural product-like molecules of over 300 compounds, 13.6% of which were identified as having adverse cellular effects. This assay provides a relatively cheap and rapid approach for identifying compounds with adverse cellular effects during screening assays, potentially reducing compound rejection due to toxicity in subsequent in vitro and in vivo assays.

  7. Elevated glucose concentration changes the content and cellular localization of AMPA receptors in the retina but not in the hippocampus.

    Science.gov (United States)

    Castilho, A F; Liberal, J T; Baptista, F I; Gaspar, J M; Carvalho, A L; Ambrósio, A F

    2012-09-06

    Diabetic retinopathy and diabetic encephalopathy are two common complications of diabetes mellitus. The impairment of glutamatergic neurotransmission in the retina and hippocampus has been suggested to be involved in the pathogenesis of these diabetic complications. In this study, we investigated the effect of elevated glucose concentration and diabetes on the protein content and surface expression of AMPA receptor subunits in the rat retina and hippocampus. We have used two models, cultured retinal and hippocampal cells exposed to elevated glucose concentration and an animal model of streptozotocin-induced type 1 diabetes. The immunoreactivity of GluA1, GluA2 and GluA4 was evaluated by Western blot and immunocytochemistry. The levels of these subunits at the plasma membrane were evaluated by biotinylation and purification of plasma membrane-associated proteins. Elevated glucose concentration increased the total levels of GluA2 subunit of AMPA receptors in retinal neural cells, but not of the subunits GluA1 or GluA4. However, at the plasma membrane, elevated glucose concentration induced an increase of all AMPA receptor subunits. In cultured hippocampal neurons, elevated glucose concentration did not induce significant alterations in the levels of AMPA receptor subunits. In the retinas of diabetic rats there were no persistent changes in the levels of AMPA receptor subunits comparing to aged-matched control retinas. Also, no consistent changes were detected in the levels of GluA1, GluA2 or GluA4 in the hippocampus of diabetic rats. We demonstrate that elevated glucose concentration induces early changes in AMPA receptor subunits, mainly in GluA2 subunit, in retinal neural cells. Conversely, hippocampal neurons seem to remain unaffected by elevated glucose concentration, concerning the expression of AMPA receptors, suggesting that AMPA receptors are more susceptible to the stress caused by elevated glucose concentration in retinal cells than in hippocampal neurons.

  8. High-content analysis of sequential events during the early phase of influenza A virus infection.

    Science.gov (United States)

    Banerjee, Indranil; Yamauchi, Yohei; Helenius, Ari; Horvath, Peter

    2013-01-01

    Influenza A virus (IAV) represents a worldwide threat to public health by causing severe morbidity and mortality every year. Due to high mutation rate, new strains of IAV emerge frequently. These IAVs are often drug-resistant and require vaccine reformulation. A promising approach to circumvent this problem is to target host cell determinants crucial for IAV infection, but dispensable for the cell. Several RNAi-based screens have identified about one thousand cellular factors that promote IAV infection. However, systematic analyses to determine their specific functions are lacking. To address this issue, we developed quantitative, imaging-based assays to dissect seven consecutive steps in the early phases of IAV infection in tissue culture cells. The entry steps for which we developed the assays were: virus binding to the cell membrane, endocytosis, exposure to low pH in endocytic vacuoles, acid-activated fusion of viral envelope with the vacuolar membrane, nucleocapsid uncoating in the cytosol, nuclear import of viral ribonucleoproteins, and expression of the viral nucleoprotein. We adapted the assays to automated microscopy and optimized them for high-content screening. To quantify the image data, we performed both single and multi-parametric analyses, in combination with machine learning. By time-course experiments, we determined the optimal time points for each assay. Our quality control experiments showed that the assays were sufficiently robust for high-content analysis. The methods we describe in this study provide a powerful high-throughput platform to understand the host cell processes, which can eventually lead to the discovery of novel anti-pathogen strategies.

  9. Remote Monitoring of Soil Water Content, Temperature, and Heat Flow Using Low-Cost Cellular (3G) IoT Technology

    Science.gov (United States)

    Ham, J. M.

    2016-12-01

    New microprocessor boards, open-source sensors, and cloud infrastructure developed for the Internet of Things (IoT) can be used to create low-cost monitoring systems for environmental research. This project describes two applications in soil science and hydrology: 1) remote monitoring of the soil temperature regime near oil and gas operations to detect the thermal signature associated with the natural source zone degradation of hydrocarbon contaminants in the vadose zone, and 2) remote monitoring of soil water content near the surface as part of a global citizen science network. In both cases, prototype data collection systems were built around the cellular (2G/3G) "Electron" microcontroller (www.particle.io). This device allows connectivity to the cloud using a low-cost global SIM and data plan. The systems have cellular connectivity in over 100 countries and data can be logged to the cloud for storage. Users can view data real time over any internet connection or via their smart phone. For both projects, data logging, storage, and visualization was done using IoT services like Thingspeak (thingspeak.com). The soil thermal monitoring system was tested on experimental plots in Colorado USA to evaluate the accuracy and reliability of different temperature sensors and 3D printed housings. The soil water experiment included comparison opens-source capacitance-based sensors to commercial versions. Results demonstrate the power of leveraging IoT technology for field research.

  10. High concentrations of H2O2 make aerobic glycolysis energetically more favourable than cellular respiration.

    Directory of Open Access Journals (Sweden)

    Hamid R Molavian

    2016-08-01

    Full Text Available Since the original observation of the Warburg Effect in cancer cells, over eight decades ago, the major question of why aerobic glycolysis is favored over oxidative phosphorylation has remained unresolved. An understanding of this phenomenon may well be the key to the development of more effective cancer therapies. In this paper, we use a semi-empirical method to throw light on this puzzle. We show that aerobic glycolysis is in fact energetically more favorable than oxidative phosphorylation for concentrations of peroxide (H2O2 above some critical threshold value. The fundamental reason for this is the activation and high engagement of the pentose phosphate pathway (PPP in response to the production of reactive oxygen species H2O2 by mitochondria and the high concentration of H2O2 (produced by mitochondria and other sources. This makes oxidative phosphorylation an inefficient source of energy since it leads (despite high levels of ATP production to a concomitant high energy consumption in order to respond to the hazardous waste products resulting from cellular processes associated with this metabolic pathway. We also demonstrate that the high concentration of H2O2 results in an increased glucose consumption, and also increases the lactate production in the case of glycolysis.

  11. Janus Separator of Polypropylene-Supported Cellular Graphene Framework for Sulfur Cathodes with High Utilization in Lithium-Sulfur Batteries.

    Science.gov (United States)

    Peng, Hong-Jie; Wang, Dai-Wei; Huang, Jia-Qi; Cheng, Xin-Bing; Yuan, Zhe; Wei, Fei; Zhang, Qiang

    2016-01-01

    Owing to the conversion chemistry of the sulfur cathode, the lithium-sulfur (Li-S) batteries exhibit high theoretical energy density. However, the intrinsic mobile redox centers during the sulfur/Li2S-to-lithium polysulfides solid-to-liquid phase transition induce low sulfur utilization and poor cycling life. Herein, the Janus separator of mesoporous cellular graphene framework (CGF)/polypropylene membrane to promote the utilization of sulfur cathode is introduced. The porous polypropylene membrane serves as an insulating substrate in contact with lithium anode while CGFs that possess high electrical conductivity of 100 S cm(-1), a large mesopore volume of 3.1 cm(3) g(-1), and a huge surface area of 2120 m(2) g(-1) are adhered on cathode side to reactivate the shuttling-back polysulfides and to preserve the ion channels. Therefore, the Li-S cell with the "two-face" CGF Janus separator exhibit a high initial capacity of 1109 mAh g(-1) and superior capacity preserved upon 800 mAh g(-1) after 250 cycles at 0.2 C, which is 40% higher on sulfur utilization efficiency than the corresponding results with routine polypropylene separators. There are significant improvements on capacity as well as electrochemical kinetics. A very high areal capacity of 5.5 mAh cm(-2) combined with high sulfur content of 80% and areal loading amount of 5.3 mg cm(-2) is achieved for such advanced configuration. The negative impact of shuttle mechanism on lowering the utilization of sulfur and overall energy density of a Li-S battery is well eliminated by applying CGF separators. Consequently, employing carbonaceous materials as Janus face of separators enlightens new opportunities for improving the utilization of active materials and energy density of devices that involve complex phase evolution and conversion electrochemistry.

  12. Janus Separator of Polypropylene‐Supported Cellular Graphene Framework for Sulfur Cathodes with High Utilization in Lithium–Sulfur Batteries

    Science.gov (United States)

    Peng, Hong‐Jie; Wang, Dai‐Wei; Cheng, Xin‐Bing; Yuan, Zhe; Wei, Fei

    2016-01-01

    Owing to the conversion chemistry of the sulfur cathode, the lithium–sulfur (Li–S) batteries exhibit high theoretical energy density. However, the intrinsic mobile redox centers during the sulfur/Li2S‐to‐lithium polysulfides solid‐to‐liquid phase transition induce low sulfur utilization and poor cycling life. Herein, the Janus separator of mesoporous cellular graphene framework (CGF)/polypropylene membrane to promote the utilization of sulfur cathode is introduced. The porous polypropylene membrane serves as an insulating substrate in contact with lithium anode while CGFs that possess high electrical conductivity of 100 S cm−1, a large mesopore volume of 3.1 cm3 g−1, and a huge surface area of 2120 m2 g−1 are adhered on cathode side to reactivate the shuttling‐back polysulfides and to preserve the ion channels. Therefore, the Li–S cell with the “two‐face” CGF Janus separator exhibit a high initial capacity of 1109 mAh g−1 and superior capacity preserved upon 800 mAh g−1 after 250 cycles at 0.2 C, which is 40% higher on sulfur utilization efficiency than the corresponding results with routine polypropylene separators. There are significant improvements on capacity as well as electrochemical kinetics. A very high areal capacity of 5.5 mAh cm−2 combined with high sulfur content of 80% and areal loading amount of 5.3 mg cm−2 is achieved for such advanced configuration. The negative impact of shuttle mechanism on lowering the utilization of sulfur and overall energy density of a Li–S battery is well eliminated by applying CGF separators. Consequently, employing carbonaceous materials as Janus face of separators enlightens new opportunities for improving the utilization of active materials and energy density of devices that involve complex phase evolution and conversion electrochemistry.

  13. A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity

    Science.gov (United States)

    Anderl, Janet L; Redpath, Stella; Ball, Andrew J

    2009-01-01

    High Content Analysis (HCA) assays combine cells and detection reagents with automated imaging and powerful image analysis algorithms, allowing measurement of multiple cellular phenotypes within a single assay. In this study, we utilized HCA to develop a novel assay for neurotoxicity. Neurotoxicity assessment represents an important part of drug safety evaluation, as well as being a significant focus of environmental protection efforts. Additionally, neurotoxicity is also a well-accepted in vitro marker of the development of neurodegenerative diseases such as Alzheimer's and Parkinson's diseases. Recently, the application of HCA to neuronal screening has been reported. By labeling neuronal cells with βIII-tubulin, HCA assays can provide high-throughput, non-subjective, quantitative measurements of parameters such as neuronal number, neurite count and neurite length, all of which can indicate neurotoxic effects. However, the role of astrocytes remains unexplored in these models. Astrocytes have an integral role in the maintenance of central nervous system (CNS) homeostasis, and are associated with both neuroprotection and neurodegradation when they are activated in response to toxic substances or disease states. GFAP is an intermediate filament protein expressed predominantly in the astrocytes of the CNS. Astrocytic activation (gliosis) leads to the upregulation of GFAP, commonly accompanied by astrocyte proliferation and hypertrophy. This process of reactive gliosis has been proposed as an early marker of damage to the nervous system. The traditional method for GFAP quantitation is by immunoassay. This approach is limited by an inability to provide information on cellular localization, morphology and cell number. We determined that HCA could be used to overcome these limitations and to simultaneously measure multiple features associated with gliosis - changes in GFAP expression, astrocyte hypertrophy, and astrocyte proliferation - within a single assay. In co

  14. A neuronal and astrocyte co-culture assay for high content analysis of neurotoxicity.

    Science.gov (United States)

    Anderl, Janet L; Redpath, Stella; Ball, Andrew J

    2009-05-05

    High Content Analysis (HCA) assays combine cells and detection reagents with automated imaging and powerful image analysis algorithms, allowing measurement of multiple cellular phenotypes within a single assay. In this study, we utilized HCA to develop a novel assay for neurotoxicity. Neurotoxicity assessment represents an important part of drug safety evaluation, as well as being a significant focus of environmental protection efforts. Additionally, neurotoxicity is also a well-accepted in vitro marker of the development of neurodegenerative diseases such as Alzheimer's and Parkinson's diseases. Recently, the application of HCA to neuronal screening has been reported. By labeling neuronal cells with betaIII-tubulin, HCA assays can provide high-throughput, non-subjective, quantitative measurements of parameters such as neuronal number, neurite count and neurite length, all of which can indicate neurotoxic effects. However, the role of astrocytes remains unexplored in these models. Astrocytes have an integral role in the maintenance of central nervous system (CNS) homeostasis, and are associated with both neuroprotection and neurodegradation when they are activated in response to toxic substances or disease states. GFAP is an intermediate filament protein expressed predominantly in the astrocytes of the CNS. Astrocytic activation (gliosis) leads to the upregulation of GFAP, commonly accompanied by astrocyte proliferation and hypertrophy. This process of reactive gliosis has been proposed as an early marker of damage to the nervous system. The traditional method for GFAP quantitation is by immunoassay. This approach is limited by an inability to provide information on cellular localization, morphology and cell number. We determined that HCA could be used to overcome these limitations and to simultaneously measure multiple features associated with gliosis - changes in GFAP expression, astrocyte hypertrophy, and astrocyte proliferation - within a single assay. In co

  15. Boron dipyrromethene (BODIPY) functionalized carbon nano-onions for high resolution cellular imaging

    Science.gov (United States)

    Bartelmess, Juergen; de Luca, Elisa; Signorelli, Angelo; Baldrighi, Michele; Becce, Michele; Brescia, Rosaria; Nardone, Valentina; Parisini, Emilio; Echegoyen, Luis; Pompa, Pier Paolo; Giordani, Silvia

    2014-10-01

    Carbon nano-onions (CNOs) are an exciting class of carbon nanomaterials, which have recently demonstrated a facile cell-penetration capability. In the present work, highly fluorescent boron dipyrromethene (BODIPY) dyes were covalently attached to the surface of CNOs. The introduction of this new carbon nanomaterial-based imaging platform, made of CNOs and BODIPY fluorophores, allows for the exploration of synergetic effects between the two building blocks and for the elucidation of its performance in biological applications. The high fluorescence intensity exhibited by the functionalized CNOs translates into an excellent in vitro probe for the high resolution imaging of MCF-7 human breast cancer cells. It was also found that the CNOs, internalized by the cells by endocytosis, localized in the lysosomes and did not show any cytotoxic effects. The presented results highlight CNOs as excellent platforms for biological and biomedical studies due to their low toxicity, efficient cellular uptake and low fluorescence quenching of attached probes.Carbon nano-onions (CNOs) are an exciting class of carbon nanomaterials, which have recently demonstrated a facile cell-penetration capability. In the present work, highly fluorescent boron dipyrromethene (BODIPY) dyes were covalently attached to the surface of CNOs. The introduction of this new carbon nanomaterial-based imaging platform, made of CNOs and BODIPY fluorophores, allows for the exploration of synergetic effects between the two building blocks and for the elucidation of its performance in biological applications. The high fluorescence intensity exhibited by the functionalized CNOs translates into an excellent in vitro probe for the high resolution imaging of MCF-7 human breast cancer cells. It was also found that the CNOs, internalized by the cells by endocytosis, localized in the lysosomes and did not show any cytotoxic effects. The presented results highlight CNOs as excellent platforms for biological and biomedical

  16. Changes and significance of SIRT3 expression in cellular senescence induced by high glucose

    Directory of Open Access Journals (Sweden)

    Bin ZHANG

    2011-09-01

    Full Text Available Objective To investigate the role of the silent information regulator 3(SIRT3 in the decrepitude process of human diploid fibroblasts(WI-38 induced by high glucose.Methods The WI-38 cells [population doublings(PDs,20-32] were cultured in media containing different concentrations of glucose as follows: low glucose(3.34mmol/L,LG,normal glucose(5.56mmol/L,NG,and high glucose(25mmol/L,HG.The protein expression levels of p21,p27,catalase,MnSOD,and SIRT3 were evaluated through Western blot.The double-label immunofluorescence assay was used to detect the location and expression of SIRT3,MnSOD,and senescence-associated heterochromatin foci(SAHF in the WI-38 cells.The ROS level was determined with fluorescent probe.Results The results from the Western blot showed that the protein expression of SIRT3,catalase,and MnSOD decreased significantly in the HG group compared with the LG and NG groups(P 0.05.SIRT3 and MnSOD were highly expressed in the cytoplasm.The ROS levels in the HG group were elevated compared with those in the LG and NG groups.Conclusion SIRT3 may play an important role in cellular senescence induced by high glucose in human diploid fibroblasts.

  17. Effects of the breed, sex and age on cellular content and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel

    Directory of Open Access Journals (Sweden)

    Giraldo Carlos E

    2013-02-01

    Full Text Available Abstract Background There is no information on the effects of the breed, gender and age on the cellular content and growth factor (GF release from equine pure-platelet rich plasma (P-PRP and pure-platelet rich gel (P-PRG. The objectives of this study were: 1 to compare the cellular composition of P-PRP with whole blood and platelet poor plasma (PPP; 2 to compare the concentration of transforming GF beta 1 (TGF-β1 and platelet derived GF isoform BB (PDGF-BB between P-PRP treated with non-ionic detergent (P-PRP+NID, P-PRG (activated with calcium gluconate -CG-, PPP+NID, PPP gel (PPG, and plasma and; 3 to evaluate and to correlate the effect of the breed, gender and age on the cellular and GF concentration for each blood component. Forty adult horses, 20 Argentinean Creole Horses (ACH and, 20 Colombian Creole Horses (CCH were included. Data were analyzed by parametric (i.e.: t-test, one way ANOVA and non parametric (Kruskal-Wallis test, Wilcoxon test tests. Correlation analysis was also performed by using the Spearman and Pearson tests. A p ≤ 0.05 was set as significant for all tests. All the blood components were compared for platelet (PLT, leukocyte (WBC, TGF-β1 and PDGF-BB concentrations. The effect of the breed, gender and age on these variables was analyzed. A P ≤ 0.05 was accepted as significant for all the tests. Results PLT counts were 1.8 and 0.6 times higher in P-PRP than in whole blood and PPP, respectively; WBC counts were 0.5 and 0.1 times lower in P-PRP, in comparison with whole blood and PPP, respectively. TGF-β1 and PDGF-BB concentrations were 2.3 and 262 times higher, respectively, in P-PRG than in plasma, and 0.59 and 0.48 times higher, respectively, in P-PRG than in PPG. P-PRG derived from CCH females or young horses presented significantly (P Conclusions Our results indicated that P-PRP obtained by a manual method was affected by intrinsic factors such as the breed, gender and age. Equine practitioners should be

  18. Cellular graphene aerogel combines ultralow weight and high mechanical strength: A highly efficient reactor for catalytic hydrogenation.

    Science.gov (United States)

    Zhang, Bingxing; Zhang, Jianling; Sang, Xinxin; Liu, Chengcheng; Luo, Tian; Peng, Li; Han, Buxing; Tan, Xiuniang; Ma, Xue; Wang, Dong; Zhao, Ning

    2016-05-12

    The construction of three-dimensional graphene aerogels (GAs) is of great importance owing to their outstanding properties for various applications. Up to now, the combination of ultralow weight and super mechanical strength for GA remains a great challenge. Here we demonstrate the fabrication of cellular GAs by a facile, easily controlled and versatile route, i.e. the chemical reduction of graphene oxide assemblies at oil-water interface under a mild condition (70 °C). The GA is ultralight (with density <3 mg cm(-3)) yet mechanically resilient because the walls of the cell closely pack in a highly ordered manner to maximize mechanical strength. The GA has been utilized as an appealing reactor for catalytic hydrogenation, which exhibited great advantages such as large oil absorption capability, exceptional catalytic activity, ease of product separation and high stability.

  19. Theoretical analyses of cellular transmembrane voltage in suspensions induced by high-frequency fields.

    Science.gov (United States)

    Zou, Yong; Wang, Changzhen; Peng, Ruiyun; Wang, Lifeng; Hu, Xiangjun

    2015-04-01

    A change of the transmembrane voltage is considered to cause biophysical and biochemical responses in cells. The present study focuses on the cellular transmembrane voltage (Δφ) induced by external fields. We detail analytical equations for the transmembrane voltage induced by external high-frequency (above the relaxation frequency of the cell membrane) fields on cells of a spherical shape in suspensions and layers. At direct current (DC) and low frequencies, the cell membrane was assumed to be non-conductive under physiologic conditions. However, with increasing frequency, the permittivity of the cytoplasm/extracellular medium and conductivity of the membrane must be accounted for. Our main work is to extend application of the analytical solution of Δφ to the high-frequency range. We first introduce the transmembrane voltage generated by DC and low-frequency exposures on a single cell. Then, we focus on cell suspensions exposed to high-frequency fields. Using the effective medium theory and the reasonable assumption, the approximate analytical solution of Δφ on cells in suspensions and layers can be derived. Phenomenological effective medium theory equations cannot be used to calculate the local electric field of cell suspensions, so we raised a possible solution based on the Bergman theory. Copyright © 2014. Published by Elsevier B.V.

  20. An objective method for High Dynamic Range source content selection

    DEFF Research Database (Denmark)

    Narwaria, Manish; Mantel, Claire; Da Silva, Matthieu Perreira

    2014-01-01

    component of such validation studies is the selection of a challenging and balanced set of source (reference) HDR content. In order to facilitate this, we present an objective method based on the premise that a more challenging HDR scene encapsulates higher contrast, and as a result will show up more...

  1. Toward high-content screening of mitochondrial morphology and membrane potential in living cells.

    Science.gov (United States)

    Iannetti, Eligio F; Willems, Peter H G M; Pellegrini, Mina; Beyrath, Julien; Smeitink, Jan A M; Blanchet, Lionel; Koopman, Werner J H

    2015-06-01

    Mitochondria are double membrane organelles involved in various key cellular processes. Governed by dedicated protein machinery, mitochondria move and continuously fuse and divide. These "mitochondrial dynamics" are bi-directionally linked to mitochondrial and cell functional state in space and time. Due to the action of the electron transport chain (ETC), the mitochondrial inner membrane displays a inside-negative membrane potential (Δψ). The latter is considered a functional readout of mitochondrial "health" and required to sustain normal mitochondrial ATP production and mitochondrial fusion. During the last decade, live-cell microscopy strategies were developed for simultaneous quantification of Δψ and mitochondrial morphology. This revealed that ETC dysfunction, changes in Δψ and aberrations in mitochondrial structure often occur in parallel, suggesting they are linked potential targets for therapeutic intervention. Here we discuss how combining high-content and high-throughput strategies can be used for analysis of genetic and/or drug-induced effects at the level of individual organelles, cells and cell populations. This article is part of a Directed Issue entitled: Energy Metabolism Disorders and Therapies.

  2. Evaluation of biological activities of highly diluted nucleotide sequences by using cellular models

    Directory of Open Access Journals (Sweden)

    Pierre Dorfman

    2012-09-01

    Full Text Available Background: highly diluted specific nucleic acids (SNA®, designed to modulate viral and cytokine genes expression, are currently used in Micro-Immunotherapy to treat viral infections and immune disorders. Although some preliminary studies have showed clinical benefit of these homeopathic preparations [1], no experimental data are available to explain their mechanism of action. Aims: to investigate the in vitro effect of two sets of highly diluted (HD SNA targeting i latent/lytic Epstein-Barr virus (SNA EBV and ii TNF-α and its receptor p55 involved in rheumatoid arthritis (SNA RA on cellular models. Methodology: serial homeopathic dilutions of SNA EBV and SNA RA (15cH-18cH were tested on a EBV-positive B-lymphoblastoid (B95-8 and on a LPS-stimulated macrophage (THP1 cell lines respectively, in comparison with agitated/diluted water and scramble DNA sequences prepared in the same conditions (negative controls. For B95-8 proliferative model, high mobility group box 1 protein (HMGB1 was used as reference. Analyzed biological parameters on B95-8 were i cell proliferation measured after 24 and 48h of incubation with HD SNA and ii expression of the EBV ZEBRA protein in response to TGF-β by Western-blotting (T+24h. For THP1 model, TNF-α synthesis and release were determined by RT-qPCR and ELISA (protein, after stimulation by LPS (1µg/ml and HD SNA co-administration. Results: we demonstrated that HD SNA RA significantly down-regulated TNF-α synthesis and release. This biological activity was showed to be specific (no effect of HD scramble SNA and related to the level of dilution (maximal effect with higher dilutions. Unexpectedly, a biological effect of agitated/diluted water was also detected in both cellular models. For B95-8 model, this effect resulted in a significant decrease of B95-8 proliferation (comparable to the HMGB1 reference and an inhibition of ZEBRA expression. Similarly, a reproducible

  3. Defining high-detail hazard maps by a cellular automata approach: application to Mount Etna (Italy

    Directory of Open Access Journals (Sweden)

    William Spataro

    2011-12-01

    Full Text Available The individuation of areas that are more likely to be affected by new events in volcanic regions is of fundamental relevance for the mitigation of the possible consequences, both in terms of loss of human life and material properties. Here, we describe a methodology for defining flexible high-detail lava-hazard maps and a technique for the validation of the results obtained. The methodology relies on: (i an accurate analysis of the past behavior of the volcano; (ii a new version of the SCIARA model for lava-flow simulation (based on the macroscopic cellular automata paradigm; and (iii high-performance parallel computing for increasing computational efficiency. The new release of the SCIARA model introduces a Bingham-like rheology as part of the minimization algorithm of the differences for the determination of outflows from a generic cell, and an improved approach to lava cooling. The method is here applied to Mount Etna, the most active volcano in Europe, and applications to land-use planning and hazard mitigation are presented.

  4. High resolution light-sheet based high-throughput imaging cytometry system enables visualization of intra-cellular organelles

    Directory of Open Access Journals (Sweden)

    Raju Regmi

    2014-09-01

    Full Text Available Visualization of intracellular organelles is achieved using a newly developed high throughput imaging cytometry system. This system interrogates the microfluidic channel using a sheet of light rather than the existing point-based scanning techniques. The advantages of the developed system are many, including, single-shot scanning of specimens flowing through the microfluidic channel at flow rate ranging from micro- to nano- lit./min. Moreover, this opens-up in-vivo imaging of sub-cellular structures and simultaneous cell counting in an imaging cytometry system. We recorded a maximum count of 2400 cells/min at a flow-rate of 700 nl/min, and simultaneous visualization of fluorescently-labeled mitochondrial network in HeLa cells during flow. The developed imaging cytometry system may find immediate application in biotechnology, fluorescence microscopy and nano-medicine.

  5. High resolution simulations of energy absorption in dynamically loaded cellular structures

    Science.gov (United States)

    Winter, R. E.; Cotton, M.; Harris, E. J.; Eakins, D. E.; McShane, G.

    2017-03-01

    Cellular materials have potential application as absorbers of energy generated by high velocity impact. CTH, a Sandia National Laboratories Code which allows very severe strains to be simulated, has been used to perform very high resolution simulations showing the dynamic crushing of a series of two-dimensional, stainless steel metal structures with varying architectures. The structures are positioned to provide a cushion between a solid stainless steel flyer plate with velocities ranging from 300 to 900 m/s, and an initially stationary stainless steel target. Each of the alternative architectures under consideration was formed by an array of identical cells each of which had a constant volume and a constant density. The resolution of the simulations was maximised by choosing a configuration in which one-dimensional conditions persisted for the full period over which the specimen densified, a condition which is most readily met by impacting high density specimens at high velocity. It was found that the total plastic flow and, therefore, the irreversible energy dissipated in the fully densified energy absorbing cell, increase (a) as the structure becomes more rodlike and less platelike and (b) as the impact velocity increases. Sequential CTH images of the deformation processes show that the flow of the cell material may be broadly divided into macroscopic flow perpendicular to the compression direction and jetting-type processes (microkinetic flow) which tend to predominate in rod and rodlike configurations and also tend to play an increasing role at increased strain rates. A very simple analysis of a configuration in which a solid flyer impacts a solid target provides a baseline against which to compare and explain features seen in the simulations. The work provides a basis for the development of energy absorbing structures for application in the 200-1000 m/s impact regime.

  6. High-resolution fiber-optic microendoscopy for in situ cellular imaging.

    Science.gov (United States)

    Pierce, Mark; Yu, Dihua; Richards-Kortum, Rebecca

    2011-01-11

    Many biological and clinical studies require the longitudinal study and analysis of morphology and function with cellular level resolution. Traditionally, multiple experiments are run in parallel, with individual samples removed from the study at sequential time points for evaluation by light microscopy. Several intravital techniques have been developed, with confocal, multiphoton, and second harmonic microscopy all demonstrating their ability to be used for imaging in situ. With these systems, however, the required infrastructure is complex and expensive, involving scanning laser systems and complex light sources. Here we present a protocol for the design and assembly of a high-resolution microendoscope which can be built in a day using off-the-shelf components for under US$5,000. The platform offers flexibility in terms of image resolution, field-of-view, and operating wavelength, and we describe how these parameters can be easily modified to meet the specific needs of the end user. We and others have explored the use of the high-resolution microendoscope (HRME) in in vitro cell culture, in excised and living animal tissues, and in human tissues in vivo. Users have reported the use of several different fluorescent contrast agents, including proflavine, benzoporphyrin-derivative monoacid ring A (BPD-MA), and fluoroscein, all of which have received full, or investigational approval from the FDA for use in human subjects. High-resolution microendoscopy, in the form described here, may appeal to a wide range of researchers working in the basic and clinical sciences. The technique offers an effective and economical approach which complements traditional benchtop microscopy, by enabling the user to perform high-resolution, longitudinal imaging in situ.

  7. An enhanced high-speed multi-digit BCD adder using quantum-dot cellular automata

    Science.gov (United States)

    Ajitha, D.; Ramanaiah, K. V.; Sumalatha, V.

    2017-02-01

    The advent of development of high-performance, low-power digital circuits is achieved by a suitable emerging nanodevice called quantum-dot cellular automata (QCA). Even though many efficient arithmetic circuits were designed using QCA, there is still a challenge to implement high-speed circuits in an optimized manner. Among these circuits, one of the essential structures is a parallel multi-digit decimal adder unit with significant speed which is very attractive for future environments. To achieve high speed, a new correction logic formulation method is proposed for single and multi-digit BCD adder. The proposed enhanced single-digit BCD adder (ESDBA) is 26% faster than the carry flow adder (CFA)-based BCD adder. The multi-digit operations are also performed using the proposed ESDBA, which is cascaded innovatively. The enhanced multi-digit BCD adder (EMDBA) performs two 4-digit and two 8-digit BCD addition 50% faster than the CFA-based BCD adder with the nominal overhead of the area. The EMDBA performs two 4-digit BCD addition 24% faster with 23% decrease in the area, similarly for 8-digit operation the EMDBA achieves 36% increase in speed with 21% less area compared to the existing carry look ahead (CLA)-based BCD adder design. The proposed multi-digit adder produces significantly less delay of (N –1) + 3.5 clock cycles compared to the N* One digit BCD adder delay required by the conventional BCD adder method. It is observed that as per our knowledge this is the first innovative proposal for multi-digit BCD addition using QCA.

  8. Impedance matching network for high frequency ultrasonic transducer for cellular applications.

    Science.gov (United States)

    Kim, Min Gon; Yoon, Sangpil; Kim, Hyung Ham; Shung, K Kirk

    2016-02-01

    An approach for the design of an impedance matching network (IMN) for high frequency ultrasonic transducers with large apertures based on impedance analysis for cellular applications is presented in this paper. The main objectives were to maximize energy transmission from the excitation source to the ultrasonic transducers for cell manipulation and to achieve low input parameters for the safe operation of an ultrasonic transducer because the piezoelectric material in high frequency ultrasonic transducers is prone to breakage due to its being extremely thin. Two ultrasonic transducers, which were made of lithium niobate single crystal with the thickness of 15 μm, having apertures of 4.3 mm (fnumber=1.23) and 2.6mm (fnumber=0.75) were tested. L-type IMN was selected for high sensitivity and compact design of the ultrasonic transducers. The target center frequency was chosen as the frequency where the electrical admittance (|Y|) and phase angle (θz) from impedance analysis was maximal and zero, respectively. The reference center frequency and reference echo magnitude were selected as the center frequency and echo magnitude, measured by pulse-echo testing, of the ultrasonic transducer without IMN. Initial component values and topology of IMN were determined using the Smith chart, and pulse-echo testing was analyzed to verify the performance of the ultrasonic transducers with and without IMN. After several iterations between changing component values and topology of IMN, and pulse-echo measurement of the ultrasonic transducer with IMN, optimized component values and topology of IMN were chosen when the measured center frequency from pulse-echo testing was comparable to the target frequency, and the measured echo magnitude was at least 30% larger than the reference echo magnitude. Performance of an ultrasonic transducer with and without IMN was tested by observing a tangible dent on the surface of a plastic petridish and single cell response after an acoustic pulse was

  9. Portal vein glucose entry triggers a coordinated cellular response that potentiates hepatic glucose uptake and storage in normal but not high-fat/high-fructose-fed dogs.

    Science.gov (United States)

    Coate, Katie C; Kraft, Guillaume; Irimia, Jose M; Smith, Marta S; Farmer, Ben; Neal, Doss W; Roach, Peter J; Shiota, Masakazu; Cherrington, Alan D

    2013-02-01

    The cellular events mediating the pleiotropic actions of portal vein glucose (PoG) delivery on hepatic glucose disposition have not been clearly defined. Likewise, the molecular defects associated with postprandial hyperglycemia and impaired hepatic glucose uptake (HGU) following consumption of a high-fat, high-fructose diet (HFFD) are unknown. Our goal was to identify hepatocellular changes elicited by hyperinsulinemia, hyperglycemia, and PoG signaling in normal chow-fed (CTR) and HFFD-fed dogs. In CTR dogs, we demonstrated that PoG infusion in the presence of hyperinsulinemia and hyperglycemia triggered an increase in the activity of hepatic glucokinase (GK) and glycogen synthase (GS), which occurred in association with further augmentation in HGU and glycogen synthesis (GSYN) in vivo. In contrast, 4 weeks of HFFD feeding markedly reduced GK protein content and impaired the activation of GS in association with diminished HGU and GSYN in vivo. Furthermore, the enzymatic changes associated with PoG sensing in chow-fed animals were abolished in HFFD-fed animals, consistent with loss of the stimulatory effects of PoG delivery. These data reveal new insight into the molecular physiology of the portal glucose signaling mechanism under normal conditions and to the pathophysiology of aberrant postprandial hepatic glucose disposition evident under a diet-induced glucose-intolerant condition.

  10. Cellular Stress Response Gene Expression During Upper and Lower Body High Intensity Exercises

    Science.gov (United States)

    Kochanowicz, Andrzej; Sawczyn, Stanisław; Niespodziński, Bartłomiej; Mieszkowski, Jan; Kochanowicz, Kazimierz

    2017-01-01

    Objectives The aim was to compare the effect of upper and lower body high-intensity exercise on chosen genes expression in athletes and non-athletes. Method Fourteen elite male artistic gymnasts (EAG) aged 20.6 ± 3.3 years and 14 physically active men (PAM) aged 19.9 ± 1.0 years performed lower and upper body 30 s Wingate Tests. Blood samples were collected before, 5 and 30 minutes after each effort to assess gene expression via PCR. Results Significantly higher mechanical parameters after lower body exercise was observed in both groups, for relative power (8.7 ± 1.2 W/kg in gymnasts, 7.2 ± 1.2 W/kg in controls, p = 0.01) and mean power (6.7 ± 0.7 W/kg in gymnasts, 5.4 ± 0.8 W/kg in controls, p = 0.01). No differences in lower versus upper body gene expression were detected for all tested genes as well as between gymnasts and physical active man. For IL-6 m-RNA time-dependent effect was observed. Conclusions Because of no significant differences in expression of genes associated with cellular stress response the similar adaptive effect to exercise may be obtained so by lower and upper body exercise. PMID:28141870

  11. Cellular lightweight concrete containing high-calcium fly ash and natural zeolite

    Science.gov (United States)

    Jitchaiyaphum, Khamphee; Sinsiri, Theerawat; Jaturapitakkul, Chai; Chindaprasirt, Prinya

    2013-05-01

    Cellular lightweight concrete (CLC) with the controlled density of approximately 800 kg/m3 was made from a preformed foam, Type-I Portland cement (OPC), fly ash (FA), or natural zeolite (NZ), and its compressive strength, setting time, water absorption, and microstructure of were tested. High-calcium FA and NZ with the median particle sizes of 14.52 and 7.72 μm, respectively, were used to partially replace OPC at 0, 10wt%, 20wt%, and 30wt% of the binder (OPC and pozzolan admixture). A water-to-binder mass ratio (W/B) of 0.5 was used for all mixes. The testing results indicated that CLC containing 10wt% NZ had the highest compressive strength. The replacement of OPC with NZ decreased the total porosity and air void size but increased the capillary porosity of the CLC. The incorporation of a suitable amount of NZ decreased the setting time, total porosity, and pore size of the paste compared with the findings with the same amount of FA. The total porosity and cumulative pore volume decreased, whereas the gel and capillary pores increased as a result of adding both pozzolans at all replacement levels. The water absorption increased as the capillary porosity increased; this effect depended on the volume of air entrained and the type or amount of pozzolan.

  12. Cellular and molecular alterations in human epithelial cells transformed by high let radiation

    Science.gov (United States)

    Hei, T. K.; Piao, C. Q.; Sutter, T.; Willey, J. C.; Suzuki, K.

    An understanding of the radiobiological effects of high LET radiation is essential for human risk estimation and radiation protection. In the present study, we show that a single, 30 cGy dose of 150 keV/mum ^4He ions can malignantly transform human papillomavirus immortalized human bronchial epithelial [BEP2D] cells. Transformed cells produce progressively growing tumors in nude mice. The transformation frequency by the single dose of alpha particles is estimated to be approximately 4 x 10^-7. Based on the average cross-sectional area of BEP2D cells, it can be calculated that a mean traversal of 1.4 particles per cell is sufficient to induce tumorigenic conversion of these cells 3 to 4 months post-irradiation. Tumorigenic BEP2D cells overexpress mutated p53 tumor suppressor oncoproteins in addition to the cell cycle control gene cyclin D1 and D2. This model provides an opportunity to study the cellular and molecular changes at the various stages in radiation carcinogenesis involving human cells.

  13. High water content in primitive continental flood basalts

    Science.gov (United States)

    Xia, Qun-Ke; Bi, Yao; Li, Pei; Tian, Wei; Wei, Xun; Chen, Han-Lin

    2016-01-01

    As the main constituent of large igneous provinces, the generation of continental flood basalts (CFB) that are characterized by huge eruption volume (>105 km3) within short time span (basaltic melts and the partition coefficient of H2O between cpx and basaltic melt. The arc-like H2O content (4.82 ± 1.00 wt.%) provides the first clear evidence that H2O plays an important role in the generation of CFB. PMID:27143196

  14. Decomposition kinetics of expanded austenite with high nitrogen contents

    DEFF Research Database (Denmark)

    Christiansen, Thomas; Somers, Marcel A. J.

    2006-01-01

    This paper addresses the decomposition kinetics of synthesized homogeneous expanded austenite formed by gaseous nitriding of stainless steel AISI 304L and AISI 316L with nitrogen contents up to 38 at.% nitrogen. Isochronal annealing experiments were carried out in both inert (N2) and reducing (H2......) atmospheres. Differential thermal analysis (DTA) and thermogravimetry were applied for identification of the decomposition reactions and X-ray diffraction analysis was applied for phase analysis. CrN precipitated upon annealing; the activation energies are 187 kJ/mol and 128 kJ/mol for AISI 316L and AISI 304L...

  15. Identifying and quantifying heterogeneity in high content analysis: application of heterogeneity indices to drug discovery.

    Directory of Open Access Journals (Sweden)

    Albert H Gough

    Full Text Available One of the greatest challenges in biomedical research, drug discovery and diagnostics is understanding how seemingly identical cells can respond differently to perturbagens including drugs for disease treatment. Although heterogeneity has become an accepted characteristic of a population of cells, in drug discovery it is not routinely evaluated or reported. The standard practice for cell-based, high content assays has been to assume a normal distribution and to report a well-to-well average value with a standard deviation. To address this important issue we sought to define a method that could be readily implemented to identify, quantify and characterize heterogeneity in cellular and small organism assays to guide decisions during drug discovery and experimental cell/tissue profiling. Our study revealed that heterogeneity can be effectively identified and quantified with three indices that indicate diversity, non-normality and percent outliers. The indices were evaluated using the induction and inhibition of STAT3 activation in five cell lines where the systems response including sample preparation and instrument performance were well characterized and controlled. These heterogeneity indices provide a standardized method that can easily be integrated into small and large scale screening or profiling projects to guide interpretation of the biology, as well as the development of therapeutics and diagnostics. Understanding the heterogeneity in the response to perturbagens will become a critical factor in designing strategies for the development of therapeutics including targeted polypharmacology.

  16. Anti-cancer agents in Saudi Arabian herbals revealed by automated high-content imaging

    KAUST Repository

    Hajjar, Dina

    2017-06-13

    Natural products have been used for medical applications since ancient times. Commonly, natural products are structurally complex chemical compounds that efficiently interact with their biological targets, making them useful drug candidates in cancer therapy. Here, we used cell-based phenotypic profiling and image-based high-content screening to study the mode of action and potential cellular targets of plants historically used in Saudi Arabia\\'s traditional medicine. We compared the cytological profiles of fractions taken from Juniperus phoenicea (Arar), Anastatica hierochuntica (Kaff Maryam), and Citrullus colocynthis (Hanzal) with a set of reference compounds with established modes of action. Cluster analyses of the cytological profiles of the tested compounds suggested that these plants contain possible topoisomerase inhibitors that could be effective in cancer treatment. Using histone H2AX phosphorylation as a marker for DNA damage, we discovered that some of the compounds induced double-strand DNA breaks. Furthermore, chemical analysis of the active fraction isolated from Juniperus phoenicea revealed possible anti-cancer compounds. Our results demonstrate the usefulness of cell-based phenotypic screening of natural products to reveal their biological activities.

  17. A High-Content Larval Zebrafish Brain Imaging Method for Small Molecule Drug Discovery

    Science.gov (United States)

    Liu, Harrison; Chen, Steven; Huang, Kevin; Kim, Jeffrey; Mo, Han; Iovine, Raffael; Gendre, Julie; Pascal, Pauline; Li, Qiang; Sun, Yaping; Dong, Zhiqiang; Arkin, Michelle; Guo, Su

    2016-01-01

    Drug discovery in whole-organisms such as zebrafish is a promising approach for identifying biologically-relevant lead compounds. However, high content imaging of zebrafish at cellular resolution is challenging due to the difficulty in orienting larvae en masse such that the cell type of interest is in clear view. We report the development of the multi-pose imaging method, which uses 96-well round bottom plates combined with a standard liquid handler to repose the larvae within each well multiple times, such that an image in a specific orientation can be acquired. We have validated this method in a chemo-genetic zebrafish model of dopaminergic neuron degeneration. For this purpose, we have developed an analysis pipeline that identifies the larval brain in each image and then quantifies neuronal health in CellProfiler. Our method achieves a SSMD* score of 6.96 (robust Z’-factor of 0.56) and is suitable for screening libraries up to 105 compounds in size. PMID:27732643

  18. Effects of Silica Aerogel Content on Microstructural and Mechanical Properties of Poly(methyl methacry-late)/Silica Aerogel Dual-scale Cellular Foams Processed in Supercritical Carbon Dioxide

    Institute of Scientific and Technical Information of China (English)

    GU Xiaoli; LUO Guoqiang; ZHANG Ruizhi; ZHANG Jian; LI Meijuan; SHEN Qiang; WANG Jin; ZHANG Lianmeng

    2016-01-01

    A novel poly(methyl-methacrylate)/silica aerogel (PMMA/SA) dual-scale cellular foam was synthesized with internal mixing followed by the supercritical carbon dioxide foaming process. The effects of silica aerogel content on the microstructural and mechanical performance of the foams were investigated by SEM, TEM analysis, and mechanical tests. The experimental results suggest that the employment of silica aerogel granule as addictive can distinctly improve the morphological feature as well as the mechanical performance in comparison to neat PMMA foam by uniformizing cell size distribution, decreasing cell size and increasing cell density. And dual-scale cells including micrometric cells of 3-10 μm and nanometric cells of about 50nm existed in the structure of foams resulting from the retained original framework structure of silica aerogel, which has not been described in other studies with the addition of various fillers. Furthermore, the mechanical strength was signiifcantly elevated even with a small amount of silica aerogel resulting from the unique microstructure, decreased cell size and enhanced cell walls. The compressive strength was 18.12 MPa and the lfexural strength was 18.90 MPa by adding 5wt% and 2wt% silica aerogel, respectively. These results demonstrate the potential to synthesize PMMA/SA dual-scale cellular foams to be used as structural materials with the advantages of low density and high strength.

  19. Multiparametric Cell Cycle Analysis Using the Operetta High-Content Imager and Harmony Software with PhenoLOGIC.

    Directory of Open Access Journals (Sweden)

    Andrew J Massey

    Full Text Available High-content imaging is a powerful tool for determining cell phenotypes at the single cell level. Characterising the effect of small molecules on cell cycle distribution is important for understanding their mechanism of action especially in oncology drug discovery but also for understanding potential toxicology liabilities. Here, a high-throughput phenotypic assay utilising the PerkinElmer Operetta high-content imager and Harmony software to determine cell cycle distribution is described. PhenoLOGIC, a machine learning algorithm within Harmony software was employed to robustly separate single cells from cell clumps. DNA content, EdU incorporation and pHH3 (S10 expression levels were subsequently utilised to separate cells into the various phases of the cell cycle. The assay is amenable to multiplexing with an additional pharmacodynamic marker to assess cell cycle changes within a specific cellular sub-population. Using this approach, the cell cycle distribution of γH2AX positive nuclei was determined following treatment with DNA damaging agents. Likewise, the assay can be multiplexed with Ki67 to determine the fraction of quiescent cells and with BrdU dual labelling to determine S-phase duration. This methodology therefore provides a relatively cheap, quick and high-throughput phenotypic method for determining accurate cell cycle distribution for small molecule mechanism of action and drug toxicity studies.

  20. High-content, high-throughput analysis of cell cycle perturbations induced by the HSP90 inhibitor XL888.

    Directory of Open Access Journals (Sweden)

    Susan K Lyman

    Full Text Available BACKGROUND: Many proteins that are dysregulated or mutated in cancer cells rely on the molecular chaperone HSP90 for their proper folding and activity, which has led to considerable interest in HSP90 as a cancer drug target. The diverse array of HSP90 client proteins encompasses oncogenic drivers, cell cycle components, and a variety of regulatory factors, so inhibition of HSP90 perturbs multiple cellular processes, including mitogenic signaling and cell cycle control. Although many reports have investigated HSP90 inhibition in the context of the cell cycle, no large-scale studies have examined potential correlations between cell genotype and the cell cycle phenotypes of HSP90 inhibition. METHODOLOGY/PRINCIPAL FINDINGS: To address this question, we developed a novel high-content, high-throughput cell cycle assay and profiled the effects of two distinct small molecule HSP90 inhibitors (XL888 and 17-AAG [17-allylamino-17-demethoxygeldanamycin] in a large, genetically diverse panel of cancer cell lines. The cell cycle phenotypes of both inhibitors were strikingly similar and fell into three classes: accumulation in M-phase, G2-phase, or G1-phase. Accumulation in M-phase was the most prominent phenotype and notably, was also correlated with TP53 mutant status. We additionally observed unexpected complexity in the response of the cell cycle-associated client PLK1 to HSP90 inhibition, and we suggest that inhibitor-induced PLK1 depletion may contribute to the striking metaphase arrest phenotype seen in many of the M-arrested cell lines. CONCLUSIONS/SIGNIFICANCE: Our analysis of the cell cycle phenotypes induced by HSP90 inhibition in 25 cancer cell lines revealed that the phenotypic response was highly dependent on cellular genotype as well as on the concentration of HSP90 inhibitor and the time of treatment. M-phase arrest correlated with the presence of TP53 mutations, while G2 or G1 arrest was more commonly seen in cells bearing wt TP53. We draw

  1. Determination of Stabiliser Contents in Advanced Gun Propellants by Reverse Phase High Performance Liquid Chromatography

    Science.gov (United States)

    1994-03-01

    HIGH PERFORMANCE LIQUID CHROMATOGRAPHY N"m A.R. TURNER AND A. WHITE...TO biEPROOU.; AND SELL THIS REPORT Determination of Stabiliser Contents in Advanced Gun Propellants by Reverse Phase High Performance Liquid Chromatography A.R...8217/......... .. Availability Cooes Dist Avaiardlo A-i Determination of Stabiliser Contents in Advanced Gun Propellants by Reverse Phase High Performance Liquid Chromatography

  2. Electric-arc synthesis of soot with high content of higher fullerenes in parallel arc

    Science.gov (United States)

    Dutlov, A. E.; Nekrasov, V. M.; Sergeev, A. G.; Bubnov, V. P.; Kareev, I. E.

    2016-12-01

    Soot with a relatively high content of higher fullerenes (C76, C78, C80, C82, C84, C86, etc.) is synthesized in a parallel arc upon evaporation of pure carbon electrodes. The content of higher fullerenes in soot extract amounts to 13.8 wt % when two electrodes are simultaneously burnt in electric-arc reactor. Such a content is comparable with the content obtained upon evaporation of composite graphite electrodes with potassium carbonate impurity.

  3. Pedagogical content knowledge and preparation of high school physics teachers

    Directory of Open Access Journals (Sweden)

    Eugenia Etkina

    2010-08-01

    Full Text Available This paper contains a scholarly description of pedagogical practices of the Rutgers Physics/Physical Science Teacher Preparation program. The program focuses on three aspects of teacher preparation: knowledge of physics, knowledge of pedagogy, and knowledge of how to teach physics (pedagogical content knowledge—PCK. The program has been in place for 7 years and has a steady production rate of an average of six teachers per year who remain in the profession. The main purpose of the paper is to provide information about a possible structure, organization, and individual elements of a program that prepares physics teachers. The philosophy of the program and the coursework can be implemented either in a physics department or in a school of education. The paper provides details about the program course work and teaching experiences and suggests ways to adapt it to other local conditions.

  4. Cellular immune profiling after sequential clofarabine and lenalidomide for high risk myelodysplastic syndromes and acute myeloid leukemia.

    Science.gov (United States)

    Jain, Prachi; Klotz, Jeffrey; Dunavin, Neil; Lu, Kit; Koklanaris, Eleftheria; Draper, Debbie; Superata, Jeanine; Chinian, Fariba; Yu, Quan; Keyvanfar, Keyvan; Wong, Susan; Muranski, Pawel; Barrett, A John; Ito, Sawa; Battiwalla, Minoo

    2017-01-01

    Patients with high risk myelodysplastic syndromes (MDS) and acute myelogenous leukemia (AML) are commonly older with multiple co-morbidities, rendering them unsuitable for intensive induction chemotherapy or transplantation. We report preliminary cellular immune profiling of four cases receiving sequential clofarabine and lenalidomide for high risk MDS and AML in a phase I study. Our results highlight the potential of immune profiling for monitoring immune-modifying agents in high risk MDS and AML.

  5. Cellular immune profiling after sequential clofarabine and lenalidomide for high risk myelodysplastic syndromes and acute myeloid leukemia

    Directory of Open Access Journals (Sweden)

    Prachi Jain

    2017-01-01

    Full Text Available Patients with high risk myelodysplastic syndromes (MDS and acute myelogenous leukemia (AML are commonly older with multiple co-morbidities, rendering them unsuitable for intensive induction chemotherapy or transplantation. We report preliminary cellular immune profiling of four cases receiving sequential clofarabine and lenalidomide for high risk MDS and AML in a phase I study. Our results highlight the potential of immune profiling for monitoring immune-modifying agents in high risk MDS and AML.

  6. A multi-channel high time resolution detector for high content imaging

    CERN Document Server

    Lapington, J S; Miller, G M; Ashton, T J R; Jarron, P; Despeisse, M; Powolny, F; Howorth, J; Milnes, J

    2009-01-01

    Medical imaging has long benefited from advances in photon counting detectors arising from space and particle physics. We describe a microchannel plate-based detector system for high content (multi-parametric) analysis, specifically designed to provide a step change in performance and throughput for measurements in imaged live cells and tissue for the ‘omics’. The detector system integrates multi-channel, high time resolution, photon counting capability into a single miniaturized detector with integrated ASIC electronics, comprising a fast, low power amplifier discriminator and TDC for every channel of the discrete pixel electronic readout, and achieving a pixel density improvement of order two magnitudes compared with current comparable devices. The device combines high performance, easy reconfigurability, and economy within a compact footprint. We present simulations and preliminary measurements in the context of our ultimate goals of 20 ps time resolution with multi-channel parallel analysis (1024 chan...

  7. High-order sliding mode control of a DC motor drive via a switched controlled multi-cellular converter

    Science.gov (United States)

    Djemaï, M.; Busawon, K.; Benmansour, K.; Marouf, A.

    2011-11-01

    In this article, we present a high-order sliding mode controller of a DC motor drive connected to a multi-cellular converter. More specifically, we design a second-order (super-twisting) control algorithm for the speed regulation of a DC motor. For this, a switching control for the multi-cellular converter is derived in order to supply the correct reference value for the speed regulation. A practical implementation of the controller is realised using a laboratory set-up. The performance and the validity of the controller are shown experimentally.

  8. In vivo effects of high-dose steroids on nucleic acid content of immunocompetent cells of renal allograft recipients

    Energy Technology Data Exchange (ETDEWEB)

    Walle, A.J.; Wong, G.Y.; Suthanthiran, M.; Rubin, A.L.; Stenzel, K.H.

    1988-03-01

    High-dose steroids administered to renal allograft recipients for treatment of acute graft rejection episodes may affect cell cycle progression of peripheral blood mononuclear (PBM) cells. DNA synthesis and cellular DNA and RNA contents of PBM cells were measured in 8 patients during clinically stable periods, and in another 10 patients both during acute rejection episodes and during 7 days of administration of high-dose steroids. Improved renal function documented successful reversal of the rejection episodes in the 10 patients. Compared with the stable patients, the rejecting patients had higher numbers of cells undergoing clonal expansion--namely, higher proportions of G1-cells and of proliferating, or S, G2, and M (SG2M) cells. Steroid treatment had no acute effects on proportions of G1 or SG2M cells in vivo or on incorporation of /sup 3/H thymidine by PBM cells in vitro. However, cells in the prereplicative compartment of the cell cycle (G0/1 cells) had significantly lower RNA content within 7 days of treatment with high doses of steroids. The results suggest that steroids do not acutely influence the posttranscriptional synthesis and the contents of nucleic acids of cells undergoing clonal expansion in vivo. The prereplicative phase of allogeneically stimulated PBM cells of renal allograft recipients may therefore be the cell cycle phase most sensitive to steroids in vivo.

  9. Thermal Analysis of ZPPR High Pu Content Stored Fuel

    Directory of Open Access Journals (Sweden)

    Charles W. Solbrig

    2014-01-01

    Full Text Available The Zero Power Physics Reactor (ZPPR operated from April 18, 1969, until 1990. ZPPR operated at low power for testing nuclear reactor designs. This paper examines the temperature of Pu content ZPPR fuel while it is in storage. Heat is generated in the fuel due to Pu and Am decay and is a concern for possible cladding damage. Damage to the cladding could lead to fuel hydriding and oxidizing. A series of computer simulations were made to determine the range of temperatures potentially occuring in the ZPPR fuel. The maximum calculated fuel temperature is 292°C (558°F. Conservative assumptions in the model intentionally overestimate temperatures. The stored fuel temperatures are dependent on the distribution of fuel in the surrounding storage compartments, the heat generation rate of the fuel, and the orientation of fuel. Direct fuel temperatures could not be measured but storage bin doors, storage sleeve doors, and storage canister temperatures were measured. Comparison of these three temperatures to the calculations indicates that the temperatures calculated with conservative assumptions are, as expected, higher than the actual temperatures. The maximum calculated fuel temperature with the most conservative assumptions is significantly below the fuel failure criterion of 600°C (1,112°F.

  10. High dynamic range images for enhancing low dynamic range content

    OpenAIRE

    Banterle, Francesco; Dellepiane, Matteo; Scopigno, Roberto

    2011-01-01

    This poster presents a practical system for enhancing the quality of Low Dynamic Range (LDR) videos using High Dynamic Range (HDR) background images. Our technique relies on the assumption that the HDR information is static in the video footage. This assumption can be valid in many scenarios where moving subjects are the main focus of the footage and do not have to interact with moving light sources or highly reflective objects. Another valid scenario is teleconferencing via webcams, where th...

  11. Social interactions of eating behaviour among high school students: a cellular automata approach.

    Science.gov (United States)

    Dabbaghian, Vahid; Mago, Vijay K; Wu, Tiankuang; Fritz, Charles; Alimadad, Azadeh

    2012-10-09

    Overweight and obesity in children and adolescents is a global epidemic posing problems for both developed and developing nations. The prevalence is particularly alarming in developed nations, such as the United States, where approximately one in three school-aged adolescents (ages 12-19) are overweight or obese. Evidence suggests that weight gain in school-aged adolescents is related to energy imbalance exacerbated by the negative aspects of the school food environment, such as presence of unhealthy food choices. While a well-established connection exists between the food environment, presently there is a lack of studies investigating the impact of the social environment and associated interactions of school-age adolescents. This paper uses a mathematical modelling approach to explore how social interactions among high school adolescents can affect their eating behaviour and food choice. In this paper we use a Cellular Automata (CA) modelling approach to explore how social interactions among school-age adolescents can affect eating behaviour, and food choice. Our CA model integrates social influences and transition rules to simulate the way individuals would interact in a social community (e.g., school cafeteria). To replicate these social interactions, we chose the Moore neighbourhood which allows all neighbours (eights cells in a two-dimensional square lattice) to influence the central cell. Our assumption is that individuals belong to any of four states; Bring Healthy, Bring Unhealthy, Purchase Healthy, and Purchase Unhealthy, and will influence each other according to parameter settings and transition rules. Simulations were run to explore how the different states interact under varying parameter settings. This study, through simulations, illustrates that students will change their eating behaviour from unhealthy to healthy as a result of positive social and environmental influences. In general, there is one common characteristic of changes across time

  12. A Study of the Efficiency of High-strength, Steel, Cellular-core Sandwich Plates in Compression

    Science.gov (United States)

    Johnson, Aldie E , Jr; Semonian, Joseph W

    1956-01-01

    Structural efficiency curves are presented for high-strength, stainless-steel, cellular-core sandwich plates of various proportions subjected to compressive end loads for temperatures of 80 F and 600 F. Optimum proportions of sandwich plates for any value of the compressive loading intensity can be determined from the curves. The efficiency of steel sandwich plates of optimum proportions is compared with the efficiency of solid plates of high-strength steel and aluminum and titanium alloys at the two temperatures.

  13. Asymmetric-detection time-stretch optical microscopy (ATOM) for ultrafast high-contrast cellular imaging in flow

    CERN Document Server

    Wong, Terence T W; Ho, Kenneth K Y; Tang, Matthew Y H; Robles, Joseph D F; Wei, Xiaoming; Chan, Antony C S; Tang, Anson H L; Lam, Edmund Y; Wong, Kenneth K Y; Chan, Godfrey C F; Shum, Ho Cheung; Tsia, Kevin K

    2013-01-01

    Accelerating imaging speed in optical microscopy is often realized at the expense of image contrast, image resolution, and detection sensitivity- a common predicament for advancing high-speed and high-throughput cellular imaging. We here demonstrate a new imaging approach, called asymmetric-detection time-stretch optical microscopy (ATOM), which can deliver ultrafast label-free high-contrast flow imaging with well delineated cellular morphological resolution and in-line optical image amplification to overcome the compromised imaging sensitivity at high speed. We show that ATOM can separately reveal the enhanced phase-gradient and absorption contrast in microfluidic live-cell imaging at a flow speed as high as ~10 m/s, corresponding to an imaging throughput of ~100,000 cells/sec. ATOM could thus be the enabling platform to meet the pressing need for intercalating optical microscopy in cellular assay, e.g. imaging flow cytometry- permitting high-throughput access to the morphological information of the individu...

  14. Asymmetric-detection time-stretch optical microscopy (ATOM) for ultrafast high-contrast cellular imaging in flow

    Science.gov (United States)

    Wong, Terence T. W.; Lau, Andy K. S.; Ho, Kenneth K. Y.; Tang, Matthew Y. H.; Robles, Joseph D. F.; Wei, Xiaoming; Chan, Antony C. S.; Tang, Anson H. L.; Lam, Edmund Y.; Wong, Kenneth K. Y.; Chan, Godfrey C. F.; Shum, Ho Cheung; Tsia, Kevin K.

    2014-01-01

    Accelerating imaging speed in optical microscopy is often realized at the expense of image contrast, image resolution, and detection sensitivity – a common predicament for advancing high-speed and high-throughput cellular imaging. We here demonstrate a new imaging approach, called asymmetric-detection time-stretch optical microscopy (ATOM), which can deliver ultrafast label-free high-contrast flow imaging with well delineated cellular morphological resolution and in-line optical image amplification to overcome the compromised imaging sensitivity at high speed. We show that ATOM can separately reveal the enhanced phase-gradient and absorption contrast in microfluidic live-cell imaging at a flow speed as high as ~10 m/s, corresponding to an imaging throughput of ~100,000 cells/sec. ATOM could thus be the enabling platform to meet the pressing need for intercalating optical microscopy in cellular assay, e.g. imaging flow cytometry – permitting high-throughput access to the morphological information of the individual cells simultaneously with a multitude of parameters obtained in the standard assay. PMID:24413677

  15. A parallel microfluidic flow cytometer for high-content screening.

    Science.gov (United States)

    McKenna, Brian K; Evans, James G; Cheung, Man Ching; Ehrlich, Daniel J

    2011-05-01

    A parallel microfluidic cytometer (PMC) uses a high-speed scanning photomultiplier-based detector to combine low-pixel-count, one-dimensional imaging with flow cytometry. The 384 parallel flow channels of the PMC decouple count rate from signal-to-noise ratio. Using six-pixel one-dimensional images, we investigated protein localization in a yeast model for human protein misfolding diseases and demonstrated the feasibility of a nuclear-translocation assay in Chinese hamster ovary (CHO) cells expressing an NFκB-EGFP reporter.

  16. High content image cytometry in the context of subnuclear organization.

    Science.gov (United States)

    De Vos, W H; Van Neste, L; Dieriks, B; Joss, G H; Van Oostveldt, P

    2010-01-01

    The organization of proteins in space and time is essential to their function. To accurately quantify subcellular protein characteristics in a population of cells with regard for the stochasticity of events in a natural context, there is a fast-growing need for image-based cytometry. Simultaneously, the massive amount of data that is generated by image-cytometric analyses, calls for tools that enable pattern recognition and automated classification. In this article, we present a general approach for multivariate phenotypic profiling of individual cell nuclei and quantification of subnuclear spots using automated fluorescence mosaic microscopy, optimized image processing tools, and supervised classification. We demonstrate the efficiency of our analysis by determination of differential DNA damage repair patterns in response to genotoxic stress and radiation, and we show the potential of data mining in pinpointing specific phenotypes after transient transfection. The presented approach allowed for systematic analysis of subnuclear features in large image data sets and accurate classification of phenotypes at the level of the single cell. Consequently, this type of nuclear fingerprinting shows potential for high-throughput applications, such as functional protein assays or drug compound screening.

  17. Organizing the Cellular and Molecular Heterogeneity in High Grade Serous Ovarian Cancer by Mass Cytometry

    Science.gov (United States)

    2015-10-01

    the vast array of genetic events found in cancer all converge on three essential cellular processes, cell fate, cell survival and genome maintenance...developed: from the Nolan Lab: Citrus 12, X-shift (unpublished), Gatefinder (unpublished), Pe’er Lab: DREMI 13. A manuscript describing X-shift has been...checked with other clustering algorithms such as used by the Citrus algorithm 12 as well as manual gating of the CyTOF data. The latter is considered

  18. Caenorhabditis elegans maintains highly compartmentalized cellular distribution of metals and steep concentration gradients of manganese.

    Directory of Open Access Journals (Sweden)

    Gawain McColl

    Full Text Available Bioinorganic chemistry is critical to cellular function. Homeostasis of manganese (Mn, for example, is essential for life. A lack of methods for direct in situ visualization of Mn and other biological metals within intact multicellular eukaryotes limits our understanding of management of these metals. We provide the first quantitative subcellular visualization of endogenous Mn concentrations (spanning two orders of magnitude associated with individual cells of the nematode, Caenorhabditis elegans.

  19. Organizing the Cellular and Molecular Heterogeneity in High-Grade Serous Ovarian Cancer by Mass Cytometry

    Science.gov (United States)

    2014-10-01

    array of genetic events found in cancer all converge on three essential cellular processes, cell fate, cell survival and genome maintenance all...Columbia) (these efforts will be ongoing throughout most of the duration of this award) New tools developed: from the Nolan Lab: Citrus [12], X-shift...p53(total) Cell cycle/ Genome integrety Lu175 Protein Biology Isotope Protein Biology Isotope CD235 Erythroid cells In113 HLA-DR DCs/B cells/monocytes

  20. High-resolution Fiber-optic Microendoscopy for in situ Cellular Imaging

    OpenAIRE

    Pierce, Mark; Yu, Dihua; Richards-Kortum, Rebecca

    2011-01-01

    Many biological and clinical studies require the longitudinal study and analysis of morphology and function with cellular level resolution. Traditionally, multiple experiments are run in parallel, with individual samples removed from the study at sequential time points for evaluation by light microscopy. Several intravital techniques have been developed, with confocal, multiphoton, and second harmonic microscopy all demonstrating their ability to be used for imaging in situ 1. With these syst...

  1. Decreased adipose tissue zinc content is associated with metabolic parameters in high fat fed Wistar rats

    OpenAIRE

    Alexey A. Tinkov; Elizaveta V. Popova; Evgenia R. Gatiatulina; Anastasia A. Skalnaya; Elena N. Yakovenko; Irina B. Alchinova; Mikhail Y. Karganov; Anatoly V. Skalny; Nikonorov, Alexandr A.

    2016-01-01

    Background. Limited data on adipose tissue zinc content in obesity exist. At the same time, the association between adipose tissue zinc content and metabolic parameters in dietary-induced obesity is poorly studied. Therefore, the primary objective of this study is to assess adipose tissue zinc content and its association  with morphometric parameters, adipokine spectrum, proinflammatory cytokines, and apolipoprotein profile in high fat fed Wistar rats. Material and method...

  2. Trend in Plutonium Content of MOX in Thermal Reactor Use and Irradiation Behavior of MOX with High Plutonium Content

    Energy Technology Data Exchange (ETDEWEB)

    Nakae, N.; Baba, T.; Kamimura, K. [Japan Nuclear Energy Safety Organization - JNES, TOKYU REIT Toranomon Bldg., 3-17-1, Toranomon, Minato-ku, Tokyo, 105-0001 (Japan); Verwerft, M.; Jutier, F. [SCK-CEN (Belgium)

    2009-06-15

    The uranium enrichment of UO{sub 2} fuel for the current power reactors, both PWR and BWR, tends to increase because of increasing burn-up target. The plutonium content of MOX fuel used in thermal reactors shall be determined in order to have reactivity worth equivalent to enriched UO{sub 2} fuel based on physical accounting method for adjusting fissile enrichment, thus the plutonium content tends to increase according to the increment of the uranium enrichment of UO{sub 2} fuel and this trend shall further be accentuated due to the fact that Pu recovered from reprocessing of the spent high burnup UO{sub 2} fuel contains less fissile isotopes. The plutonium content is calculated by use of the physical accounting method with the plutonium having several kinds of isotope ratios and the calculation results indicate that the plutonium content in MOX will evolve to ratios in excess of 10%. It shall be, therefore, important to know the irradiation behavior of MOX with high plutonium content of more than 10 wt%. MOX fuel rods having a plutonium content of about 14 wt% and fabricated by use of MIMAS process have been irradiated under PWR conditions in the Belgian test reactors BR-3 and BR-2. The peak fuel rod burn-up of the fuel rods studied in this paper ranges from 31 to 37 GWd/t-HM, and their average burnup is about 22-26 GWd/t-HM with the rod averaged linear heat generation rate of about 15-21 kW/m. The MOX rods are investigated by destructive and non-destructive post irradiation examinations and some of them are now continued to be irradiated in BR-2. Mixed Oxide (U,Pu)O{sub 2} fuel produced by the MIMAS process results in a fine dispersion of Pu enriched particles in a UO{sub 2} matrix and effectively gives three enrichment classes: low, medium and high enriched. The high enriched particles (often called 'Pu spots'), have an enrichment of around 25 wt% Pu, the low enriched phase is the UO{sub 2} matrix and contains only trace amounts of Pu. An

  3. Bioprinting-Based High-Throughput Fabrication of Three-Dimensional MCF-7 Human Breast Cancer Cellular Spheroids

    Directory of Open Access Journals (Sweden)

    Kai Ling

    2015-06-01

    Full Text Available Cellular spheroids serving as three-dimensional (3D in vitro tissue models have attracted increasing interest for pathological study and drug-screening applications. Various methods, including microwells in particular, have been developed for engineering cellular spheroids. However, these methods usually suffer from either destructive molding operations or cell loss and non-uniform cell distribution among the wells due to two-step molding and cell seeding. We have developed a facile method that utilizes cell-embedded hydrogel arrays as templates for concave well fabrication and in situ MCF-7 cellular spheroid formation on a chip. A custom-built bioprinting system was applied for the fabrication of sacrificial gelatin arrays and sequentially concave wells in a high-throughput, flexible, and controlled manner. The ability to achieve in situ cell seeding for cellular spheroid construction was demonstrated with the advantage of uniform cell seeding and the potential for programmed fabrication of tissue models on chips. The developed method holds great potential for applications in tissue engineering, regenerative medicine, and drug screening.

  4. Synthesis and Characterization of Mesoporous Silicon Oxynitride MCM-41 with High Nitrogen Content

    Institute of Scientific and Technical Information of China (English)

    ZHANG Cunman; XU Zheng; LIU Qian

    2005-01-01

    Mesoporous silicon oxynitrides MCM-41 were synthesized successfully. The resulting materials not only have high nitrogen contents and good structural characteristics of MCM-41 (high surface area, narrow pore size distribution and good order), but also are amorphous. The composition and structure of the materials were investigated by CNH element analysis, XPS, Si MAS NMR, XRD, HRTEM and N2 sorption, respectively. Mesoporous silicon oxynitrides MCM-41 with a high nitrogen content are still non-crystal (amorphous).

  5. Exploiting the multiplexing capabilities of tandem mass tags for high-throughput estimation of cellular protein abundances by mass spectrometry.

    Science.gov (United States)

    Ahrné, Erik; Martinez-Segura, Amalia; Syed, Afzal Pasha; Vina-Vilaseca, Arnau; Gruber, Andreas J; Marguerat, Samuel; Schmidt, Alexander

    2015-09-01

    The generation of dynamic models of biological processes critically depends on the determination of precise cellular concentrations of biomolecules. Measurements of system-wide absolute protein levels are particularly valuable information in systems biology. Recently, mass spectrometry based proteomics approaches have been developed to estimate protein concentrations on a proteome-wide scale. However, for very complex proteomes, fractionation steps are required, increasing samples number and instrument analysis time. As a result, the number of full proteomes that can be routinely analyzed is limited. Here we combined absolute quantification strategies with the multiplexing capabilities of isobaric tandem mass tags to determine cellular protein abundances in a high throughput and proteome-wide scale even for highly complex biological systems, such as a whole human cell line. We generated two independent data sets to demonstrate the power of the approach regarding sample throughput, dynamic range, quantitative precision and accuracy as well as proteome coverage in comparison to existing mass spectrometry based strategies.

  6. Method of preparing a high solids content, low viscosity ceramic slurry

    Science.gov (United States)

    Tiegs, Terry N.; Wittmer, Dale E.

    1995-01-01

    A method for producing a high solids content, low viscosity ceramic slurry composition comprises turbomilling a dispersion of a ceramic powder in a liquid to form a slurry having a viscosity less than 100 centipoise and a solids content equal to or greater than 48 volume percent.

  7. Content-Based Curriculum for High-Ability Learners, Second Edition

    Science.gov (United States)

    VanTassel-Baska, Joyce, Ed.; Little, Catherine A., Ed.

    2011-01-01

    The newly updated "Content-Based Curriculum for High-Ability Learners" provides a solid introduction to curriculum development in gifted and talented education. Written by experts in the field of gifted education, this text uses cutting-edge design techniques and aligns the core content with national and state standards. In addition to a revision…

  8. Composition and Content of High-Molecular-Weight Glutenin Subunits and Their Effects on Wheat Quality

    Institute of Scientific and Technical Information of China (English)

    SONG Jian-min; LIU Ai-feng; WU Xiang-yun; LIU Jian-jun; ZHAO Zhen-dong; LIU Guang-tian

    2002-01-01

    Sedimentation values, flour glutenin macropolymer (GMP) contents, composition and contents of high-molecular-weight (HMW) glutenin subunits (GS) of 233 flour samples were determined. Our data indicated that subunit 1 occurred more frequently at Glu-A1, subunit pair 7 + 8 at Glu- B1 and 2 + 12 at Glu-D1. The significant relationships between Glu-1 quality score and total HMW glutenin content, sedimentation value and GMP content suggested that the composition of HMW-GS affects wheat quality strongly. Moreover,the total content of HMW-GS was correlated with certain quality parameters more significantly. Relationship between subunit 5 + 10 content and breadmaking quality was better than others, but 2 + 12, 7 + 8, 7 + 9 and 4 + 12 also correlated with certain quality parameters significantly. The contents of total HMW-glutenin, x-type subunits and y-type subunits related with sedimentation value, flour GMP content, and Glu-1 quality score more strongly than that of individual subunit or subunit pair. The flour GMP content, with excellent correlation to sedimentation value, total contents of HMW glutenin, x- and y-type subunits and many other quality parameters, could be an ideal indicator of breadmaking quality at earlier generations for breeding purpose for its simple procedure and small scale.

  9. High Seas High Schoolers: Creating ERESE Content on an Expedition to Samoa

    Science.gov (United States)

    Delaney, R.; English, B.; Staudigel, D.; Staudigel, H.; Koppers, A.; Hart, S.

    2005-12-01

    As part of the ERESE program, three high school seniors aboard the ALIA expedition generated contents and published a live trip website with a wide range of information about the science and personal aspects of the cruise. These activities served to relate as much meaningful information about the month-long research cruise in the South Pacific as possible, to people of all ages and skills. The website http://earthref.org/ERESE/projects/ALIA/ has reports on almost every aspect of the research cruise; from what it is like staying on the Research Vessel Kilo Moana for a month, to operating machinery, to interviews with the captain and chief scientists, and even how the equipment aboard works, in less than technical terms. An effective way to relay what was actually going on aboard the Kilo Moana, were the daily reports, written by the high school students, complete with the pictures and videos taken that day. This website connected the ALIA cruise to high school students and classrooms, who were following the expedition through the website both in the United States and in Samoa. High school seniors designed and implemented the "CruiseWatch" feature on the Alia website. This "applet" extracts data from the shipboard datastream and relays them to the ERESE website at the San Diego Supercomputing center via satellite, where they are prepared for real-time display on the cruise website. Data displayed include the ships' location on the map, geographic coordinates, heading, speed and wind speed. During dredging operations it displays the length of wire deployed, wire speed and wire tension. Overall the website with the daily reports and photographs, in addition to other web media, gave this trip a unique ability to engage people from around the world in researching oceanographic and geological phenomena.

  10. Adapting human pluripotent stem cells to high-throughput and high-content screening.

    Science.gov (United States)

    Desbordes, Sabrina C; Studer, Lorenz

    2013-01-01

    The increasing use of human pluripotent stem cells (hPSCs) as a source of cells for drug discovery, cytotoxicity assessment and disease modeling requires their adaptation to large-scale culture conditions and screening formats. Here, we describe a simple and robust protocol for the adaptation of human embryonic stem cells (hESCs) to high-throughput screening (HTS). This protocol can also be adapted to human induced pluripotent stem cells (hiPSCs) and high-content screening (HCS). We also describe a 7-d assay to identify compounds with an effect on hESC self-renewal and differentiation. This assay can be adapted to a variety of applications. The procedure involves the culture expansion of hESCs, their adaptation to 384-well plates, the addition of small molecules or other factors, and finally data acquisition and processing. In this protocol, the optimal number of hESCs plated in 384-well plates has been adapted to HTS/HCS assays of 7 d.

  11. Rapid Directional Solidification with Ultra-High Temperature Gradient and Cellular Spacing Selection of Cu-Mn Alloy

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    The detailed laser surface remelting experiments of Cu-31.4 wt pct Mn and Cu-26.6 wt pct Mn alloys on a 5 kW CO2laser were carried out to study the effects of processing parameters (scanning velocity, output power of laser) on the growthdirection of microstructure in the molten pool and cellular spacing selection under the condition of ultra-high temperaturegradient and rapid directional solidification. The experimental results show that the growth direction of microstructure isstrongly affected by laser processing parameters. The ultra-high temperature gradient directional solidification can be realizedon the surface of samples during laser surface remelting by controlling laser processing parameters, the temperature gradientand growth velocity can reach 106 K/m and 24.1 mm/s, respectively, and the solidification microstructure in the center ofthe molten pool grows along the laser beam scanning direction. There exists a distribution range of cellular spacings underthe laser rapid solidification conditions, and the average spacing decreases with increasing of growth rate. The maximum,λ minimum, λmin, and average primary spacing,λ, as functions of growth rate, Vb, can be given by, λ =12.54V b-0.61,λmin=4.47 Vb-0.52, λ=9.09Vb-0.62, respectively. The experimental results are compared with the current Hunt-Lu model forrapid cellular/dendritic growth, and a good agreement is found.

  12. Changes in the topography of cellular components in pea root statocytes exposed to high gradient magnetic fields

    Science.gov (United States)

    Belyavskaya, Ninel A.; Polishchuk, Olexandr V.; Kondrachuk, Alexander V.

    2005-08-01

    High-gradient magnetic field (HGMF) is one of methods, by which gravitropism in plants is studied. The aim of our study was elucidation of HGMF effects on topography of cellular components in root statocytes of 4- day Pisum sativum L. seedlings in comparison to gravistimulation. Under gravistimulation during 5, 30 and 60 min seedlings were rotated 45o; magnetostimulation was carried out along gap between two NdFeB magnets (0.7 T). Morphometric measurements were made from images of whole statocytes, for upper, middle and lower thirds of cells, and proximal and distal halves of cells. Morphometric analysis revealed that HGMF resulted in the redistribution of all cellular components in statocytes. The correlation in the amyloplast distribution between gravistimulation and magnetostimulation was established.

  13. REQUIREMENT OF FLUIDITY OF HIGH WATER CONTENT MATERIALS FORTHE GETWAY-SIDE BACKFILLING TECHNIQUE

    Institute of Scientific and Technical Information of China (English)

    QiTaiyue; MaNianjie

    1996-01-01

    Through analyzing the effects of water consumption, diameter of solid particle, and flow velocity on the fluidity of high water content material slurry, the relationship among the fluidity, the isotropy of the slurry, and the pumping facilities applied in getway-side backfilling has been found. And the requirment of fluidity of high water content material for the design of getway-side backfilling technique is put forward in the paper.

  14. A degradation model for high kitchen waste content municipal solid waste.

    Science.gov (United States)

    Chen, Yunmin; Guo, Ruyang; Li, Yu-Chao; Liu, Hailong; Zhan, Tony Liangtong

    2016-12-01

    Municipal solid waste (MSW) in developing countries has a high content of kitchen waste (KW), and therefore contains large quantities of water and non-hollocellulose degradable organics. The degradation of high KW content MSW cannot be well simulated by the existing degradation models, which are mostly established for low KW content MSW in developed countries. This paper presents a two-stage anaerobic degradation model for high KW content MSW with degradations of hollocellulose, sugars, proteins and lipids considered. The ranges of the proportions of chemical compounds in MSW components are summarized with the recommended values given. Waste components are grouped into rapidly or slowly degradable categories in terms of the degradation rates under optimal water conditions for degradation. In the proposed model, the unionized VFA inhibitions of hydrolysis/acidogenesis and methanogenesis are considered as well as the pH inhibition of methanogenesis. Both modest and serious VFA inhibitions can be modeled by the proposed model. Default values for the parameters in the proposed method can be used for predictions of degradations of both low and high KW content MSW. The proposed model was verified by simulating two laboratory experiments, in which low and high KW content MSW were used, respectively. The simulated results are in good agreement with the measured data of the experiments. The results show that under low VFA concentrations, the pH inhibition of methanogenesis is the main inhibition to be considered, while the inhibitions of both hydrolysis/acidogenesis and methanogenesis caused by unionized VFA are significant under high VFA concentrations. The model is also used to compare the degradation behaviors of low and high KW content MSW under a favorable environmental condition, and it shows that the gas potential of high KW content MSW releases more quickly.

  15. Enhanced ethanol fermentation by engineered Saccharomyces cerevisiae strains with high spermidine contents.

    Science.gov (United States)

    Kim, Sun-Ki; Jo, Jung-Hyun; Jin, Yong-Su; Seo, Jin-Ho

    2017-05-01

    Construction of robust and efficient yeast strains is a prerequisite for commercializing a biofuel production process. We have demonstrated that high intracellular spermidine (SPD) contents in Saccharomyces cerevisiae can lead to improved tolerance against various fermentation inhibitors, including furan derivatives and acetic acid. In this study, we examined the potential applicability of the S. cerevisiae strains with high SPD contents under two cases of ethanol fermentation: glucose fermentation in repeated-batch fermentations and xylose fermentation in the presence of fermentation inhibitors. During the sixteen times of repeated-batch fermentations using glucose as a sole carbon source, the S. cerevisiae strains with high SPD contents maintained higher cell viability and ethanol productivities than a control strain with lower SPD contents. Specifically, at the sixteenth fermentation, the ethanol productivity of a S. cerevisiae strain with twofold higher SPD content was 31% higher than that of the control strain. When the SPD content was elevated in an engineered S. cerevisiae capable of fermenting xylose, the resulting S. cerevisiae strain exhibited much 40-50% higher ethanol productivities than the control strain during the fermentations of synthetic hydrolysate containing high concentrations of fermentation inhibitors. These results suggest that the strain engineering strategy to increase SPD content is broadly applicable for engineering yeast strains for robust and efficient production of ethanol.

  16. Development of a New Foxtail Millet Germplasm with Super Early Maturity and High Iron Content

    Institute of Scientific and Technical Information of China (English)

    LIU Zheng-li; SUN Shi-xian; CHENG Ru-hong; HUANG Wen-sheng; LIU Jun-xin; QU Zhu-feng; XIA Xue-yan; SHI Zhi-gang

    2006-01-01

    Super Early Maturation No.2 is a new foxtail millet germplasm developed by using the targeted-character-gene-bankbreeding method. It has several outstanding characteristics. (1) Super early maturity. It can normally mature in Bashang,Hebei Province, China, where the altitude is around 1 400 meters, and it needs 1 650℃ of effective accumulated temperature in the growing period. (2) Millet is rich in iron. The iron content of the millet of Super Early Maturation No.2 is 54.10 mg kg-1,which is 62.0% higher than the average iron content of the foxtail millet varieties in China. (3) High crude fat content. Its crude fat content is 6.24%, which is 54.1% higher than the average content of foxtail millet varieties in China. (4) High crude protein content. The average of the crude protein content of all the foxtail millet varieties in China is 12.71%, only5% varieties surpass 14%, however, the crude protein content of Super Early Maturation No.2 is 14.36%, which is rare in improved varieties of foxtail millet in China. (5) Overall characteristics are good.

  17. High-content screening of yeast mutant libraries by shotgun lipidomics

    DEFF Research Database (Denmark)

    Tarasov, Kirill; Stefanko, Adam; Casanovas, Albert;

    2014-01-01

    To identify proteins with a functional role in lipid metabolism and homeostasis we designed a high-throughput platform for high-content lipidomic screening of yeast mutant libraries. To this end, we combined culturing and lipid extraction in 96-well format, automated direct infusion nanoelectrosp......To identify proteins with a functional role in lipid metabolism and homeostasis we designed a high-throughput platform for high-content lipidomic screening of yeast mutant libraries. To this end, we combined culturing and lipid extraction in 96-well format, automated direct infusion...... factor KAR4 precipitated distinct lipid metabolic phenotypes. These results demonstrate that the high-throughput shotgun lipidomics platform is a valid and complementary proxy for high-content screening of yeast mutant libraries....

  18. High-performance full adder architecture in quantum-dot cellular automata

    Directory of Open Access Journals (Sweden)

    Hamid Rashidi

    2017-06-01

    Full Text Available Quantum-dot cellular automata (QCA is a new and promising computation paradigm, which can be a viable replacement for the complementary metal–oxide–semiconductor technology at nano-scale level. This technology provides a possible solution for improving the computation in various computational applications. Two QCA full adder architectures are presented and evaluated: a new and efficient 1-bit QCA full adder architecture and a 4-bit QCA ripple carry adder (RCA architecture. The proposed architectures are simulated using QCADesigner tool version 2.0.1. These architectures are implemented with the coplanar crossover approach. The simulation results show that the proposed 1-bit QCA full adder and 4-bit QCA RCA architectures utilise 33 and 175 QCA cells, respectively. Our simulation results show that the proposed architectures outperform most results so far in the literature.

  19. Cellular scaling rules for the brain of Artiodactyla include a highly folded cortex with few neurons

    Directory of Open Access Journals (Sweden)

    Rodrigo eSiqueira Kazu

    2014-11-01

    Full Text Available Quantitative analysis of the cellular composition of rodent, primate, insectivore and afrotherian brains has shown that nonneuronal scaling rules are similar across these mammalian orders that diverged about 95 million years ago, and therefore appear to be conserved in evolution, while neuronal scaling rules appear to be free to vary in a clade-specific manner. Here we analyze the cellular scaling rules that apply to the brain of artiodactyls, a group within the order Cetartiodactyla, believed to be a relatively recent radiation from the common Eutherian ancestor. We find that artiodactyls share nonneuronal scaling rules with all groups analyzed previously. Artiodactyls share with afrotherians and rodents, but not with primates, the neuronal scaling rules that apply to the cerebral cortex and cerebellum. The neuronal scaling rules that apply to the remaining brain areas are however distinct in artiodactyls. Importantly, we show that the folding index of the cerebral cortex scales with the number of neurons in the cerebral cortex in distinct fashions across artiodactyls, afrotherians, rodents, and primates, such that the artiodactyl cerebral cortex is more convoluted than primate cortices of similar numbers of neurons. Our findings suggest that the scaling rules found to be shared across modern afrotherians, glires and artiodactyls applied to the common Eutherian ancestor, such as the relationship between the mass of the cerebral cortex as a whole and its number of neurons. In turn, the distribution of neurons along the surface of the cerebral cortex, which is related to its degree of gyrification, appears to be a clade-specific characteristic. If the neuronal scaling rules for artiodactyls extend to all cetartiodactyls, we predict that the large cerebral cortex of cetaceans will still have fewer neurons than the human cerebral cortex.

  20. Cellular scaling rules for the brain of Artiodactyla include a highly folded cortex with few neurons.

    Science.gov (United States)

    Kazu, Rodrigo S; Maldonado, José; Mota, Bruno; Manger, Paul R; Herculano-Houzel, Suzana

    2014-01-01

    Quantitative analysis of the cellular composition of rodent, primate, insectivore, and afrotherian brains has shown that non-neuronal scaling rules are similar across these mammalian orders that diverged about 95 million years ago, and therefore appear to be conserved in evolution, while neuronal scaling rules appear to be free to vary in a clade-specific manner. Here we analyze the cellular scaling rules that apply to the brain of artiodactyls, a group within the order Cetartiodactyla, believed to be a relatively recent radiation from the common Eutherian ancestor. We find that artiodactyls share non-neuronal scaling rules with all groups analyzed previously. Artiodactyls share with afrotherians and rodents, but not with primates, the neuronal scaling rules that apply to the cerebral cortex and cerebellum. The neuronal scaling rules that apply to the remaining brain areas are, however, distinct in artiodactyls. Importantly, we show that the folding index of the cerebral cortex scales with the number of neurons in the cerebral cortex in distinct fashions across artiodactyls, afrotherians, rodents, and primates, such that the artiodactyl cerebral cortex is more convoluted than primate cortices of similar numbers of neurons. Our findings suggest that the scaling rules found to be shared across modern afrotherians, glires, and artiodactyls applied to the common Eutherian ancestor, such as the relationship between the mass of the cerebral cortex as a whole and its number of neurons. In turn, the distribution of neurons along the surface of the cerebral cortex, which is related to its degree of gyrification, appears to be a clade-specific characteristic. If the neuronal scaling rules for artiodactyls extend to all cetartiodactyls, we predict that the large cerebral cortex of cetaceans will still have fewer neurons than the human cerebral cortex.

  1. Impact of image segmentation on high-content screening data quality for SK-BR-3 cells

    Directory of Open Access Journals (Sweden)

    Li Yizheng

    2007-09-01

    Full Text Available Abstract Background High content screening (HCS is a powerful method for the exploration of cellular signalling and morphology that is rapidly being adopted in cancer research. HCS uses automated microscopy to collect images of cultured cells. The images are subjected to segmentation algorithms to identify cellular structures and quantitate their morphology, for hundreds to millions of individual cells. However, image analysis may be imperfect, especially for "HCS-unfriendly" cell lines whose morphology is not well handled by current image segmentation algorithms. We asked if segmentation errors were common for a clinically relevant cell line, if such errors had measurable effects on the data, and if HCS data could be improved by automated identification of well-segmented cells. Results Cases of poor cell body segmentation occurred frequently for the SK-BR-3 cell line. We trained classifiers to identify SK-BR-3 cells that were well segmented. On an independent test set created by human review of cell images, our optimal support-vector machine classifier identified well-segmented cells with 81% accuracy. The dose responses of morphological features were measurably different in well- and poorly-segmented populations. Elimination of the poorly-segmented cell population increased the purity of DNA content distributions, while appropriately retaining biological heterogeneity, and simultaneously increasing our ability to resolve specific morphological changes in perturbed cells. Conclusion Image segmentation has a measurable impact on HCS data. The application of a multivariate shape-based filter to identify well-segmented cells improved HCS data quality for an HCS-unfriendly cell line, and could be a valuable post-processing step for some HCS datasets.

  2. Magnetic Resonance Cell-Tracking Studies: Spectrophotometry-Based Method for the Quantification of Cellular Iron Content after Loading with Superparamagnetic Iron Oxide Nanoparticles

    Directory of Open Access Journals (Sweden)

    Ingrid Böhm

    2011-07-01

    Full Text Available The purpose of this article is to present a user-friendly tool for quantifying the iron content of superparamagnetic labeled cells before cell tracking by magnetic resonance imaging (MRI. Iron quantification was evaluated by using Prussian blue staining and spectrophotometry. White blood cells were labeled with superparamagnetic iron oxide (SPIO nanoparticles. Labeling was confirmed by light microscopy. Subsequently, the cells were embedded in a phantom and scanned on a 3 T magnetic resonance tomography (MRT whole-body system. Mean peak wavelengths Λpeak was determined at A720nm (range 719–722 nm. Linearity was proven for the measuring range 0.5 to 10 μg Fe/mL (r = .9958; p = 2.2 × 10−12. The limit of detection was 0.01 μg Fe/mL (0.1785 mM, and the limit of quantification was 0.04 μg Fe/mL (0.714 mM. Accuracy was demonstrated by comparison with atomic absorption spectrometry. Precision and robustness were also proven. On T2-weighted images, signal intensity varied according to the iron concentration of SPIO-labeled cells. Absorption spectrophotometry is both a highly sensitive and user-friendly technique that is feasible for quantifying the iron content of magnetically labeled cells. The presented data suggest that spectrophotometry is a promising tool for promoting the implementation of magnetic resonance-based cell tracking in routine clinical applications (from bench to bedside.

  3. Cellular-level near-wall unsteadiness of high-hematocrit erythrocyte flow using confocal {mu}PIV

    Energy Technology Data Exchange (ETDEWEB)

    Patrick, Michael J. [Carnegie Mellon University, Molecular Biosensor and Imaging Center (MBIC), Pittsburgh, PA (United States); Chen, Chia-Yuan; Dur, Onur; Pekkan, Kerem [Carnegie Mellon University, Department of Biomedical and Mechanical Engineering, Pittsburgh, PA (United States); Frakes, David H. [Arizona State University, School of Biological and Health Systems Engineering and School of Electrical, Computer, and Energy Engineering, Tempe, AZ (United States)

    2011-04-15

    In hemodynamics, the inherent intermittency of two-phase cellular-level flow has received little attention. Unsteadiness is reported and quantified for the first time in the literature using a combination of fluorescent dye labeling, time-resolved scanning confocal microscopy, and micro-particle image velocimetry ({mu}PIV). The near-wall red blood cell (RBC) motion of physiologic high-hematocrit blood in a rectangular microchannel was investigated under pressure-driven flow. Intermittent flow was associated with (1) the stretching of RBCs as they passed through RBC clusters with twisting motions; (2) external flow through local obstacles; and (3) transitionary rouleaux formations. Velocity profiles are presented for these cases. Unsteady flow clustered in local regions. Extra-cellular fluid flow generated by individual RBCs was examined using submicron fluorescent microspheres. The capabilities of confocal {mu}PIV post-processing were verified using synthetic raw PIV data for validation. Cellular interactions and oscillating velocity profiles are presented, and 3D data are made available for computational model validation. (orig.)

  4. Automated high-content live animal drug screening using C. elegans expressing the aggregation prone serpin α1-antitrypsin Z.

    Directory of Open Access Journals (Sweden)

    Sager J Gosai

    Full Text Available The development of preclinical models amenable to live animal bioactive compound screening is an attractive approach to discovering effective pharmacological therapies for disorders caused by misfolded and aggregation-prone proteins. In general, however, live animal drug screening is labor and resource intensive, and has been hampered by the lack of robust assay designs and high throughput work-flows. Based on their small size, tissue transparency and ease of cultivation, the use of C. elegans should obviate many of the technical impediments associated with live animal drug screening. Moreover, their genetic tractability and accomplished record for providing insights into the molecular and cellular basis of human disease, should make C. elegans an ideal model system for in vivo drug discovery campaigns. The goal of this study was to determine whether C. elegans could be adapted to high-throughput and high-content drug screening strategies analogous to those developed for cell-based systems. Using transgenic animals expressing fluorescently-tagged proteins, we first developed a high-quality, high-throughput work-flow utilizing an automated fluorescence microscopy platform with integrated image acquisition and data analysis modules to qualitatively assess different biological processes including, growth, tissue development, cell viability and autophagy. We next adapted this technology to conduct a small molecule screen and identified compounds that altered the intracellular accumulation of the human aggregation prone mutant that causes liver disease in α1-antitrypsin deficiency. This study provides powerful validation for advancement in preclinical drug discovery campaigns by screening live C. elegans modeling α1-antitrypsin deficiency and other complex disease phenotypes on high-content imaging platforms.

  5. Resolution of the cellular proteome of the nucleocapsid protein from a highly pathogenic isolate of porcine reproductive and respiratory syndrome virus identifies PARP-1 as a cellular target whose interaction is critical for virus biology.

    Science.gov (United States)

    Liu, Long; Lear, Zoe; Hughes, David J; Wu, Weining; Zhou, En-min; Whitehouse, Adrian; Chen, Hongying; Hiscox, Julian A

    2015-03-23

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a major threat to the swine industry and food security worldwide. The nucleocapsid (N) protein is a major structural protein of PRRSV. The primary function of this protein is to encapsidate the viral RNA genome, and it is also thought to participate in the modulation of host cell biology and recruitment of cellular factors to facilitate virus infection. In order to the better understand these latter roles the cellular interactome of PRRSV N protein was defined using label free quantitative proteomics. This identified several cellular factors that could interact with the N protein including poly [ADP-ribose] polymerase 1 (PARP-1), a cellular protein, which can add adenosine diphosphate ribose to a protein. Use of the PARP-1 small molecule inhibitor, 3-AB, in PRRSV infected cells demonstrated that PARP-1 was required and acted as an enhancer factor for virus biology. Serial growth of PRRSV in different concentrations of 3-AB did not yield viruses that were able to grow with wild type kinetics, suggesting that by targeting a cellular protein crucial for virus biology, resistant phenotypes did not emerge. This study provides further evidence that cellular proteins, which are critical for virus biology, can also be targeted to ablate virus growth and provide a high barrier for the emergence of drug resistance. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Effect of high temperature and excessive light on glutathione content in apple peel

    Institute of Scientific and Technical Information of China (English)

    Jianguang ZHANG; Shaochun CHEN; Yingli LI; Bao DI; Jianqiang ZHANG; Yufang LIU

    2008-01-01

    The present experiment was conducted to examine the changing patterns of glutathione (GSH) contents in apple peel stressed by high temperature and excessive solar radiation. By comparing the parameters of temperature and light conditions where fruits grow with the GSH contents in them, the mutual relationship was disclosed. Meanwhile, the changes of antioxidant capacity of fruits stressed at different levels were studied under artificially controlled conditions in a laboratory. Also, the effect of applying various types of exogenous substances was evaluated on endogenous GSH contents. The results indicated that within a certain range, a positive correlation was found between the stressed extents to which fruits were subject by high temperature and exces-sive light, and GSH contents in fruit peel. Moreover, fruits on southwest (SW) exposure contained a very significantly higher amount of GSH than those on other exposures. In laboratory experiments, it was proved that fruit GSH contents increased with temperatures within a certain range but they declined beyond a given limit. The temperature rising modes had a great influence on fruit GSH contents, and gradual tem-perature increase was favorable to an increase of antioxidant capability in fruit peel, thus providing a theoretical basis for exerting appropriate acclimation on fruits. It was also docu-mented that application of four exogenous formulations could significantly increase the endogenous GSH contents, among which AsA (ascorbit acid)+BA (benzoic acid) and SA (sali-cylic acid) treatments exhibited better results, 33.97% and 31.81% higher than the control, respectively.

  7. A High-Precision Micropipette Sensor for Cellular-Level Real-Time Thermal Characterization

    Directory of Open Access Journals (Sweden)

    Wonseok Chang

    2011-09-01

    Full Text Available We report herein development of a novel glass micropipette thermal sensor fabricated in a cost-effective manner, which is capable of measuring steady thermal fluctuation at spatial resolution of ~2 µm with an accuracy of ±0.01 °C. We produced and tested various micrometer-sized sensors, ranging from 2 µm to 30 µm. The sensor comprises unleaded low-melting-point solder alloy (Sn-based as a core metal inside a pulled borosilicate glass pipette and a thin film of nickel coating outside, creating a thermocouple junction at the tip. The sensor was calibrated using a thermally insulated calibration chamber, the temperature of which can be controlled with an accuracy of ±0.01 °C, and the thermoelectric power (Seebeck coefficient of the sensor was recorded from 8.46 to 8.86 µV/°C. We have demonstrated the capability of measuring temperatures at a cellular level by inserting our temperature sensor into the membrane of a live retinal pigment epithelium cell subjected to a laser beam with a focal spot of 6 μm. We measured transient temperature profiles and the maximum temperatures were in the range of 38–55 ± 0.5 °C.

  8. Lipolysis - a highly regulated multi-enzyme complex mediates the catabolism of cellular fat stores.

    Science.gov (United States)

    Lass, Achim; Zimmermann, Robert; Oberer, Monika; Zechner, Rudolf

    2011-01-01

    Lipolysis is the biochemical pathway responsible for the catabolism of triacylglycerol (TAG) stored in cellular lipid droplets. The hydrolytic cleavage of TAG generates non-esterified fatty acids, which are subsequently used as energy substrates, essential precursors for lipid and membrane synthesis, or mediators in cell signaling processes. Consistent with its central importance in lipid and energy homeostasis, lipolysis occurs in essentially all tissues and cell types, it is most abundant, however, in white and brown adipose tissue. Over the last 5years, important enzymes and regulatory protein factors involved in lipolysis have been identified. These include an essential TAG hydrolase named adipose triglyceride lipase (ATGL) [annotated as patatin-like phospholipase domain-containing protein A2], the ATGL activator comparative gene identification-58 [annotated as α/β hydrolase containing protein 5], and the ATGL inhibitor G0/G1 switch gene 2. Together with the established hormone-sensitive lipase [annotated as lipase E] and monoglyceride lipase, these proteins constitute the basic "lipolytic machinery". Additionally, a large number of hormonal signaling pathways and lipid droplet-associated protein factors regulate substrate access and the activity of the "lipolysome". This review summarizes the current knowledge concerning the enzymes and regulatory processes governing lipolysis of fat stores in adipose and non-adipose tissues. Special emphasis will be given to ATGL, its regulation, and physiological function. Copyright © 2010 Elsevier Ltd. All rights reserved.

  9. Lipolysis – A highly regulated multi-enzyme complex mediates the catabolism of cellular fat stores

    Science.gov (United States)

    Lass, Achim; Zimmermann, Robert; Oberer, Monika; Zechner, Rudolf

    2011-01-01

    Summary Lipolysis is the biochemical pathway responsible for the catabolism of triacylglycerol (TAG) stored in cellular lipid droplets. The hydrolytic cleavage of TAG generates non-esterified fatty acids, which are subsequently used as energy substrates, essential precursors for lipid and membrane synthesis, or mediators in cell signaling processes. Consistent with its central importance in lipid and energy homeostasis, lipolysis occurs in essentially all tissues and cell types, it is most abundant, however, in white and brown adipose tissue. Over the last 5 years, important enzymes and regulatory protein factors involved in lipolysis have been identified. These include an essential TAG hydrolase named adipose triglyceride lipase (ATGL) [annotated as patatin-like phospholipase domain-containing protein A2], the ATGL activator comparative gene identification-58 [annotated as α/β hydrolase containing protein 5], and the ATGL inhibitor G0/G1 switch gene 2. Together with the established hormone-sensitive lipase [annotated as lipase E] and monoglyceride lipase, these proteins constitute the basic “lipolytic machinery”. Additionally, a large number of hormonal signaling pathways and lipid droplet-associated protein factors regulate substrate access and the activity of the “lipolysome”. This review summarizes the current knowledge concerning the enzymes and regulatory processes governing lipolysis of fat stores in adipose and non-adipose tissues. Special emphasis will be given to ATGL, its regulation, and physiological function. PMID:21087632

  10. Optimisation of rubberised concrete with high rubber content: an experimental investigation

    OpenAIRE

    Raffoul, S.; Garcia, R; Pilakoutas, K.; Guadagnini, M.; Flores Medina, N.

    2016-01-01

    This article investigates experimentally the behaviour of rubberised concrete (RuC) with high rubber content so as to fully utilise the mechanical properties of vulcanised rubber. The fresh properties and short-term uniaxial compressive strength of 40 rubberised concrete mixes were assessed. The parameters examined included the volume (0–100%) and type of mineral aggregate replacement (fine or coarse), water or admixture contents, type of binder, rubber particle properties, and rubber surface...

  11. Investigations of DNA damage induction and repair resulting from cellular exposure to high dose-rate pulsed proton beams

    Energy Technology Data Exchange (ETDEWEB)

    Renis, M.; Malfa, G.; Tomasello, B. [Drug Sciences Department, University of Catania, Catania (Italy); Borghesi, M.; Schettino, G. [Queen' s University Belfast, Northern Ireland (United Kingdom); Favetta, M.; Romano, F.; Cirrone, G. A. P. [National Institute for Nuclear Physics (INFN-LNS), Catania (Italy); Manti, L. [Physics Science Department, University of Naples Federico II, Naples, and National Institute for Nuclear Physics (INFN), Naples (Italy)

    2013-07-26

    Studies regarding the radiobiological effects of low dose radiation, microbeam irradiation services have been developed in the world and today laser acceleration of protons and heavy ions may be used in radiation therapy. The application of different facilities is essential for studying bystander effects and relating signalling phenomena in different cells or tissues. In particular the use of ion beams results advantageous in cancer radiotherapy compared to more commonly used X-rays, since the ability of ions in delivering lethal amount of doses into the target tumour avoiding or limiting damage to the contiguous healthy tissues. At the INFN-LNS in Catania, a multidisciplinary radiobiology group is strategically structured aimed to develop radiobiological research, finalised to therapeutic applications, compatible with the use of high dose laser-driven ion beams. The characteristic non-continuous dose rates with several orders of magnitude of laser-driven ion beams makes this facility very interesting in the cellular systems' response to ultra-high dose rates with non-conventional pulse time intervals cellular studies. Our group have projected to examine the effect of high dose laser-driven ion beams on two cellular types: foetal fibroblasts (normal control cells) and DU145 (prostate cancer cells), studying the modulation of some different bio-molecular parameters, in particular cell proliferation and viability, DNA damage, redox cellular status, morphological alterations of both the cytoskeleton components and some cell organelles and the possible presence of apoptotic or necrotic cell death. Our group performed preliminary experiments with high energy (60 MeV), dose rate of 10 Gy/min, doses of 1, 2, 3 Gy and LET 1 keV/μm on human foetal fibroblasts (control cells). We observed that cell viability was not influenced by the characteristics of the beam, the irradiation conditions or the analysis time. Conversely, DNA damage was present at time 0, immediately

  12. Bringing the light to high throughput screening: use of optogenetic tools for the development of recombinant cellular assays

    Science.gov (United States)

    Agus, Viviana; Di Silvio, Alberto; Rolland, Jean Francois; Mondini, Anna; Tremolada, Sara; Montag, Katharina; Scarabottolo, Lia; Redaelli, Loredana; Lohmer, Stefan

    2015-03-01

    The use of light-activated proteins represents a powerful tool to control biological processes with high spatial and temporal precision. These so called "optogenetic" technologies have been successfully validated in many recombinant systems, and have been widely applied to the study of cellular mechanisms in intact tissues or behaving animals; to do that, complex, high-intensity, often home-made instrumentations were developed to achieve the optimal power and precision of light stimulation. In our study we sought to determine if this optical modulation can be obtained also in a miniaturized format, such as a 384-well plate, using the instrumentations normally dedicated to fluorescence analysis in High Throughput Screening (HTS) activities, such as for example the FLIPR (Fluorometric Imaging Plate Reader) instrument. We successfully generated optogenetic assays for the study of different ion channel targets: the CaV1.3 calcium channel was modulated by the light-activated Channelrhodopsin-2, the HCN2 cyclic nucleotide gated (CNG) channel was modulated by the light activated bPAC adenylyl cyclase, and finally the genetically encoded voltage indicator ArcLight was efficiently used to measure potassium, sodium or chloride channel activity. Our results showed that stable, robust and miniaturized cellular assays can be developed using different optogenetic tools, and efficiently modulated by the FLIPR instrument LEDs in a 384-well format. The spatial and temporal resolution delivered by this technology might enormously advantage the early stages of drug discovery, leading to the identification of more physiological and effective drug molecules.

  13. High content image-based screening of a protease inhibitor library reveals compounds broadly active against Rift Valley fever virus and other highly pathogenic RNA viruses.

    Directory of Open Access Journals (Sweden)

    Rajini Mudhasani

    2014-08-01

    Full Text Available High content image-based screening was developed as an approach to test a protease inhibitor small molecule library for antiviral activity against Rift Valley fever virus (RVFV and to determine their mechanism of action. RVFV is the causative agent of severe disease of humans and animals throughout Africa and the Arabian Peninsula. Of the 849 compounds screened, 34 compounds exhibited ≥ 50% inhibition against RVFV. All of the hit compounds could be classified into 4 distinct groups based on their unique chemical backbone. Some of the compounds also showed broad antiviral activity against several highly pathogenic RNA viruses including Ebola, Marburg, Venezuela equine encephalitis, and Lassa viruses. Four hit compounds (C795-0925, D011-2120, F694-1532 and G202-0362, which were most active against RVFV and showed broad-spectrum antiviral activity, were selected for further evaluation for their cytotoxicity, dose response profile, and mode of action using classical virological methods and high-content imaging analysis. Time-of-addition assays in RVFV infections suggested that D011-2120 and G202-0362 targeted virus egress, while C795-0925 and F694-1532 inhibited virus replication. We showed that D011-2120 exhibited its antiviral effects by blocking microtubule polymerization, thereby disrupting the Golgi complex and inhibiting viral trafficking to the plasma membrane during virus egress. While G202-0362 also affected virus egress, it appears to do so by a different mechanism, namely by blocking virus budding from the trans Golgi. F694-1532 inhibited viral replication, but also appeared to inhibit overall cellular gene expression. However, G202-0362 and C795-0925 did not alter any of the morphological features that we examined and thus may prove to be good candidates for antiviral drug development. Overall this work demonstrates that high-content image analysis can be used to screen chemical libraries for new antivirals and to determine their

  14. High and Low Consensus Groups: A Content and Relational Interaction Analysis.

    Science.gov (United States)

    DeStephen, Rolayne S.

    1983-01-01

    Analyzed the complete interaction of high and low consensus groups in a basic small group course. Interaction analysis indicated that both the relational and content levels of communication are significantly different for high versus low consensus groups. The conclusion that increased feedback leads to decision satisfaction was confirmed. (JAC)

  15. Innovation management and marketing in the high-tech sector: A content analysis of advertisements

    DEFF Research Database (Denmark)

    Gerhard, D.; Brem, Alexander; Baccarella, Ch.

    2011-01-01

    Advertizing high-technology products is a tricky and critical task for every company, since it means operating in an environment with high market uncertainty. The work presents results of a content analysis of 110 adverts for consumer electronics products which examines how these products and the...

  16. Evaluation of Compatibility of ToxCast High-Throughput/High-Content Screening Assays with Engineered Nanomaterials

    Science.gov (United States)

    High-throughput and high-content screens are attractive approaches for prioritizing nanomaterial hazards and informing targeted testing due to the impracticality of using traditional toxicological testing on the large numbers and varieties of nanomaterials. The ToxCast program a...

  17. Calculation Method to Determine the Group Composition of Vacuum Distillate with High Content of Saturated Hydrocarbons

    Directory of Open Access Journals (Sweden)

    Nazarova Galina

    2016-01-01

    Full Text Available Calculation method to determine the group composition of the heavy fraction of vacuum distillate with high content of saturated hydrocarbons, obtained by vacuum distillation of the residue from the West Siberian oil with subsequent hydrotreating, are given in this research. The method is built on the basis of calculation the physico-chemical characteristics and the group composition of vacuum distillate according to the fractional composition and density considering with high content of saturated hydrocarbons in the fraction. Calculation method allows to determine the content of paraffinic, naphthenic, aromatic hydrocarbons and the resins in vacuum distillate with high accuracy and can be used in refineries for rapid determination of the group composition of vacuum distillate.

  18. Graph cut and image intensity-based splitting improves nuclei segmentation in high-content screening

    Science.gov (United States)

    Farhan, Muhammad; Ruusuvuori, Pekka; Emmenlauer, Mario; Rämö, Pauli; Yli-Harja, Olli; Dehio, Christoph

    2013-02-01

    Quantification of phenotypes in high-content screening experiments depends on the accuracy of single cell analysis. In such analysis workflows, cell nuclei segmentation is typically the first step and is followed by cell body segmentation, feature extraction, and subsequent data analysis workflows. Therefore, it is of utmost importance that the first steps of high-content analysis are done accurately in order to guarantee correctness of the final analysis results. In this paper, we present a novel cell nuclei image segmentation framework which exploits robustness of graph cut to obtain initial segmentation for image intensity-based clump splitting method to deliver the accurate overall segmentation. By using quantitative benchmarks and qualitative comparison with real images from high-content screening experiments with complicated multinucleate cells, we show that our method outperforms other state-of-the-art nuclei segmentation methods. Moreover, we provide a modular and easy-to-use implementation of the method for a widely used platform.

  19. Optimization of the Content of Tricalcium Silicate of High Cementing Clinker

    Institute of Scientific and Technical Information of China (English)

    CHEN Lin; SHEN Xiaodong; MA Suhua; HUANG Yeping; ZHONG Baiqian

    2011-01-01

    Optimization of the content of tricalcium silicate (C3S) of high cementing clinker was investigated. The content of free-CaO(f-CaO), mineral composite, the content of C3S in the clinker and the hydration product were analyzed by chemical analysis and X-ray diffraction (XRD). "K Value" method of QXRD was selected as a quantitative analysis way to measure the content of C3S, and the strength of cement paste was determined. The results show that at a water cement ratio of 0.29, the strength of cement paste with 73% C3S can be up to 97.5 MPa at 28 days age. The strength at 28 d of cement with 73% C3S is 16% higher than that with 78% C3S at water requirement for normal consistency. The relationship between the strength of high cementing Portland cement and the content of C3S in the clinker is nonlinear. According to the strength of cement paste, the optimal content of C3S in cement clinker is around 73% in this paper.

  20. Decreased adipose tissue zinc content is associated with metabolic parameters in high fat fed Wistar rats

    Directory of Open Access Journals (Sweden)

    Alexey A. Tinkov

    2016-03-01

    Full Text Available Background. Limited data on adipose tissue zinc content in obesity exist. At the same time, the association between adipose tissue zinc content and metabolic parameters in dietary-induced obesity is poorly studied. Therefore, the primary objective of this study is to assess adipose tissue zinc content and its association  with morphometric parameters, adipokine spectrum, proinflammatory cytokines, and apolipoprotein profile in high fat fed Wistar rats. Material and methods. A total of 48 adult female Wistar rats were used in the present study. Rats were fed either control (10% of fat or high fat diet (31.6% of fat. Adipose tissue zinc content was assessed using inductively coupled plasma mass spectrometry. Rats’ serum was examined for adiponectin, leptin, insulin, interleukin-6, and tumor necrosis factor-α using enzyme-linked immunosorbent assay kits. Serum glucose and apolipoprotein spectrum were also evaluated. Results. High fat feeding resulted in a significant 34% decrease in adipose tissue zinc content in comparison to the control values. Fat pad zinc levels were significantly inversely associated with morphometric param- eters, circulating leptin, insulin, tumor necrosis factor-α levels and HOMA-IR values. At the same time,      a significant correlation with apolipoprotein A1 concentration was observed. Conclusion. Generally, the obtained data indicate that (1 high fat feeding results in decreased adipose tis- sue zinc content; (2 adipose tissue zinc content is tightly associated with excessive adiposity, inflammation, insulin resistance and potentially atherogenic changes.

  1. Just Working with the Cellular Machine: A High School Game for Teaching Molecular Biology

    Science.gov (United States)

    Cardoso, Fernanda Serpa; Dumpel, Renata; Gomes da Silva, Luisa B.; Rodrigues, Carlos R.; Santos, Dilvani O.; Cabral, Lucio Mendes; Castro, Helena C.

    2008-01-01

    Molecular biology is a difficult comprehension subject due to its high complexity, thus requiring new teaching approaches. Herein, we developed an interdisciplinary board game involving the human immune system response against a bacterial infection for teaching molecular biology at high school. Initially, we created a database with several…

  2. Magnetic and mechanical properties of FeSi alloys with high Si content

    Institute of Scientific and Technical Information of China (English)

    LU Feng-shuang; QIAO Liang; BI Xiao-fang

    2006-01-01

    The chemical vapor(CVD) deposition-diffusion method was applied to prepare FeSi alloys with high silicon content up to 6.5%. In spite of various deposition and post-annealing, the sample remains α-Fe bcc structure. The cross section of the composition was analyzed to evaluate the Si content and distribution before and after annealing. The results show that the soft magnetic properties are improved by increasing the silicon content. For the samples containing about 6.5% Si, the coercivity decreases to 60 from 237.3 A/m of the original. It is also obtained that, in addition to the Si content, Si distribution has a large influence on the core loss due to the effect of resistivity. The micro-hardnesses were also evaluated along the cross-section after various annealings.

  3. Cytoskeletal re-arrangement in TGF-β1-induced alveolar epithelial-mesenchymal transition studied by atomic force microscopy and high-content analysis.

    Science.gov (United States)

    Buckley, Stephen T; Medina, Carlos; Davies, Anthony M; Ehrhardt, Carsten

    2012-04-01

    Epithelial-mesenchymal transition (EMT) is closely implicated in the pathogenesis of idiopathic pulmonary fibrosis. Associated with this phenotypic transition is the acquisition of an elongated cell morphology and establishment of stress fibers. The extent to which these EMT-associated changes influence cellular mechanics is unclear. We assessed the biomechanical properties of alveolar epithelial cells (A549) following exposure to TGF-β1. Using atomic force microscopy, changes in cell stiffness and surface membrane features were determined. Stimulation with TGF-β1 gave rise to a significant increase in stiffness, which was augmented by a collagen I matrix. Additionally, TGF-β1-treated cells exhibited a rougher surface profile with notable protrusions. Simultaneous quantitative examination of the morphological attributes of stimulated cells using an image-based high-content analysis system revealed dramatic alterations in cell shape, F-actin content and distribution. Together, these investigations point to a strong correlation between the cytoskeletal-associated cellular architecture and the mechanical dynamics of alveolar epithelial cells undergoing EMT. From the Clinical Editor: Epithelial-mesenchymal transition is implicated in the pathogenesis of pulmonary fibrosis. Using atomic force microscopy, the authors demonstrate a strong correlation between the cytoskeletal-associated cellular architecture and the mechanical dynamics of alveolar epithelial cells undergoing mesenchymal transition.

  4. Enzymatic Hydrolysis of biomasses having a high dry matter (DM) content

    DEFF Research Database (Denmark)

    2010-01-01

    The present invention relates to a process for liquefaction and saccharification of polysaccharide containing biomasses, having a relatively high dry matter content. The present invention combines enzymatic hydrolysis with a type of mixing relying on the principle of gravity ensuring that the bio......The present invention relates to a process for liquefaction and saccharification of polysaccharide containing biomasses, having a relatively high dry matter content. The present invention combines enzymatic hydrolysis with a type of mixing relying on the principle of gravity ensuring...

  5. Multi Feature Content Based Video Retrieval Using High Level Semantic Concept

    Directory of Open Access Journals (Sweden)

    Hamdy K. Elminir

    2012-07-01

    Full Text Available Content-based retrieval allows finding information by searching its content rather than its attributes. The challenge facing content-based video retrieval (CBVR is to design systems that can accurately and automatically process large amounts of heterogeneous videos. Moreover, content-based video retrieval system requires in its first stage to segment the video stream into separate shots. Afterwards features are extracted for video shots representation. And finally, choose a similarity/distance metric and an algorithm that is efficient enough to retrieve query - related videos results. There are two main issues in this process; the first is how to determine the best way for video segmentation and key frame selection. The second is the features used for video representation. Various features can be extracted for this sake including either low or high level features. A key issue is how to bridge the gap between low and high level features. This paper proposes a system for a content based video retrieval system that tries to address the aforementioned issues by using adaptive threshold for video segmentation and key frame selection as well as using both low level features together with high level semantic object annotation for video representation. Experimental results show that the use of multi features increases both precision and recall rates by about 13% to 19 % than traditional system that uses only color feature for video retrieval.

  6. Use of a Machine Learning-Based High Content Analysis Approach to Identify Photoreceptor Neurite Promoting Molecules.

    Science.gov (United States)

    Fuller, John A; Berlinicke, Cynthia A; Inglese, James; Zack, Donald J

    2016-01-01

    High content analysis (HCA) has become a leading methodology in phenotypic drug discovery efforts. Typical HCA workflows include imaging cells using an automated microscope and analyzing the data using algorithms designed to quantify one or more specific phenotypes of interest. Due to the richness of high content data, unappreciated phenotypic changes may be discovered in existing image sets using interactive machine-learning based software systems. Primary postnatal day four retinal cells from the photoreceptor (PR) labeled QRX-EGFP reporter mice were isolated, seeded, treated with a set of 234 profiled kinase inhibitors and then cultured for 1 week. The cells were imaged with an Acumen plate-based laser cytometer to determine the number and intensity of GFP-expressing, i.e. PR, cells. Wells displaying intensities and counts above threshold values of interest were re-imaged at a higher resolution with an INCell2000 automated microscope. The images were analyzed with an open source HCA analysis tool, PhenoRipper (Rajaram et al., Nat Methods 9:635-637, 2012), to identify the high GFP-inducing treatments that additionally resulted in diverse phenotypes compared to the vehicle control samples. The pyrimidinopyrimidone kinase inhibitor CHEMBL-1766490, a pan kinase inhibitor whose major known targets are p38α and the Src family member lck, was identified as an inducer of photoreceptor neuritogenesis by using the open-source HCA program PhenoRipper. This finding was corroborated using a cell-based method of image analysis that measures quantitative differences in the mean neurite length in GFP expressing cells. Interacting with data using machine learning algorithms may complement traditional HCA approaches by leading to the discovery of small molecule-induced cellular phenotypes in addition to those upon which the investigator is initially focusing.

  7. High contents of very long-chain polyunsaturated fatty acids in different moss species.

    Science.gov (United States)

    Beike, Anna K; Jaeger, Carsten; Zink, Felix; Decker, Eva L; Reski, Ralf

    2014-02-01

    Polyunsaturated fatty acids (PUFAs) are important cellular compounds with manifold biological functions. Many PUFAs are essential for the human diet and beneficial for human health. In this study, we report on the high amounts of very long-chain (vl) PUFAs (≥C₂₀) such as arachidonic acid (AA) in seven moss species. These species were established in axenic in vitro culture, as a prerequisite for comparative metabolic studies under highly standardized laboratory conditions. In the model organism Physcomitrella patens, tissue-specific differences in the fatty acid compositions between the filamentous protonema and the leafy gametophores were observed. These metabolic differences correspond with differential gene expression of fatty acid desaturase (FADS)-encoding genes in both developmental stages, as determined via microarray analyses. Depending on the developmental stage and the species, AA amounts for 6-31 %, respectively, of the total fatty acids. Subcellular localization of the corresponding FADS revealed the endoplasmic reticulum as the cellular compartment for AA synthesis. Our results show that vlPUFAs are highly abundant metabolites in mosses. Standardized cultivation techniques using photobioreactors along with the availability of the P. patens genome sequence and the high rate of homologous recombination are the basis for targeted metabolic engineering in moss. The potential of producing vlPUFAs of interest from mosses will be highlighted as a promising area in plant biotechnology.

  8. Innovation management and marketing in the high-tech sector: A content analysis of advertisements

    DEFF Research Database (Denmark)

    Gerhard, D.; Brem, Alexander; Baccarella, Ch.

    2011-01-01

    Advertizing high-technology products is a tricky and critical task for every company, since it means operating in an environment with high market uncertainty. The work presents results of a content analysis of 110 adverts for consumer electronics products which examines how these products...... and the incorporated technology are communicated to the customer. Based on established coding schemes, the content and the appeals of the adverts are evaluated by coders. The results show that these adverts are very informative, mainly have rational appeals and feature products being in the early stage of their life...... cycles. Regarding the specific content, the ‘mother brand’ is shown to play a very important role. On the other hand, the results show that incorporated technology, its superiority and functionality do not play an important role in the advertisements...

  9. The influence of cellular structures on flow stress of high strength components manufactured using SLM

    DEFF Research Database (Denmark)

    Mahshid, Rasoul; Hansen, Hans Nørgaard; Loft Højbjerre, Klaus

    2016-01-01

    Additive manufacturing has shown significant improvement in material and machines for high-quality solid freeform fabrication processes such as selective laser melting (SLM). In particular, manufacturing lattice structures using the SLM procedure is of interest. This research examines the effect...

  10. Physiological conditions conducive to high cell density and high cyanophycin content in Ralstonia eutropha strain H16 possessing a KDPG aldolase gene-dependent addiction system.

    Science.gov (United States)

    Lin, Kaichien; Elbahloul, Yasser; Steinbüchel, Alexander

    2012-03-01

    The recombinant strain of Ralstonia eutropha H16-PHB(-)4-∆eda (pBBR1MCS-2::cphA (6308)/eda (H16)) presenting a 2-keto-3-desoxy-phosphogluconate (KDPG) aldolase (eda) gene-dependent catabolic addiction system for plasmid maintenance when using gluconate or fructose as sole carbon source was used in this study. The effects of the initial pH, the nitrogen-to-carbon ratio, the inorganic components of medium, the oxygen supply, and the different carbon and nitrogen sources on the cell dry matter (CDM) and the cyanophycin granule polypeptide (CGP) content of the cells were studied in a mineral salts medium (MSM) without any additional amino acids or CGP precursor substrates. The experiments were designed to systematically find out the optimal conditions for growth of cells to high densities and for high CGP contents of the cells. Maximum contents of water-insoluble CGP and water-soluble CGP, contributing to 47.5% and 5.8% (w/w) of CDM, respectively, were obtained at the 30-L scale cultivation when cells were cultivated in MSM medium containing sufficient supplements of fructose, NH(3), K(2)SO(4), MgSO(4)[Symbol: see text]7H(2)O, Fe(Ш)NH(4)-citrate, CaCl(2)[Symbol: see text]2H(2)O, and trace elements (SL6). The molecular masses of water-insoluble and water-soluble CGP ranged from 25 to 31 kDa and from 15 to 21 kDa, respectively. High cell densities of up to 82.8 g CDM/L containing up to 37.8% (w/w) water-insoluble CGP at the 30-L scale cultivation were also obtained. This is by far the best combination of high cell density and high cellular CGP contents ever reported, and it showed that efficient production of CGP at the industrial scale in white biotechnology could be achieved.

  11. A quantitative proteomic analysis of cellular responses to high glucose media in Chinese hamster ovary cells.

    Science.gov (United States)

    Liu, Zhenke; Dai, Shujia; Bones, Jonathan; Ray, Somak; Cha, Sangwon; Karger, Barry L; Li, Jingyi Jessica; Wilson, Lee; Hinckle, Greg; Rossomando, Anthony

    2015-01-01

    A goal in recombinant protein production using Chinese hamster ovary (CHO) cells is to achieve both high specific productivity and high cell density. Addition of glucose to the culture media is necessary to maintain both cell growth and viability. We varied the glucose concentration in the media from 5 to 16 g/L and found that although specific productivity of CHO-DG44 cells increased with the glucose level, the integrated viable cell density decreased. To examine the biological basis of these results, we conducted a discovery proteomic study of CHO-DG44 cells grown under batch conditions in normal (5 g/L) or high (15 g/L) glucose over 3, 6, and 9 days. Approximately 5,000 proteins were confidently identified against an mRNA-based CHO-DG44 specific proteome database, with 2,800 proteins quantified with at least two peptides. A self-organizing map algorithm was used to deconvolute temporal expression profiles of quantitated proteins. Functional analysis of altered proteins suggested that differences in growth between the two glucose levels resulted from changes in crosstalk between glucose metabolism, recombinant protein expression, and cell death, providing an overall picture of the responses to high glucose environment. The high glucose environment may enhance recombinant dihydrofolate reductase in CHO cells by up-regulating NCK1 and down-regulating PRKRA, and may lower integrated viable cell density by activating mitochondrial- and endoplasmic reticulum-mediated cell death pathways by up-regulating HtrA2 and calpains. These proteins are suggested as potential targets for bioengineering to enhance recombinant protein production.

  12. Antihyperlipidemic and hepatoprotective effects of Gardenin A in cellular and high fat diet fed rodent models.

    Science.gov (United States)

    Toppo, Erenius; Darvin, S Sylvester; Esakkimuthu, S; Stalin, A; Balakrishna, K; Sivasankaran, K; Pandikumar, P; Ignacimuthu, S; Al-Dhabi, N A

    2017-05-01

    The gum of Gardenia resinifera Roth., is one of the important drugs used in the Indian system of medicine and a source of unique polymethoxylated flavones. This study was aimed to evaluate the antihyperlipidemic and anti-NAFLD effects of Gardenin A (Gar-A) from G. resinifera gum using in vitro and in vivo models. Gar-A was isolated from G. resinifera gum and was identified on the basis of the physical and spectral data. Toxicity of Gar-A to HepG2 cells was evaluated using MTT assay. The ability of Gar-A to reduce steatosis was assessed using oleate-palmitate induced HepG2 cell lines by estimating the lipid levels by ORO staining and by estimating the intracellular triglyceride content. Effect of Gar-A on amelioration of lipotoxicity was measured by estimating the LDH levels. The doses for in vivo experiments were fixed by Irwin test, between 50 and 100 mg/kg concentrations, through oral route. The acute antihyperlipidemic effect of Gar-A was assessed in Triton WR-1339 induced hyperlipidemic animals. The chronic antihyperlipidemic and anti-NAFLD effects of Gar-A were evaluated in HFD fed rats. In vitro experiments with HepG2 cell line indicated that the cells treated with Gar-A did not show any significant reduction in the viability up to 70 μg/mL concentration. Steatotic HepG2 cells treated with Gar-A showed a significant reduction in lipid accumulation at 2.5-10 μg/mL concentrations. In triton induced hyperlipidemic rats, the treatment significantly reduced the lipid levels at the synthesis phase. The treatment with Gar-A to the HFD fed animals significantly lowered the steatosis and transaminase levels. The other biochemical parameters such as TC, TG, LDL-c, ALP and ACP were also decreased significantly. Treatment with Gar-A significantly lowered the hyperlipidemia and fat accumulation in the liver; detailed molecular investigations are necessary to establish the antihyperlipidemic and hepatoprotective potentials of Gar-A. Copyright © 2017 Elsevier B

  13. Cellular automata

    CERN Document Server

    Codd, E F

    1968-01-01

    Cellular Automata presents the fundamental principles of homogeneous cellular systems. This book discusses the possibility of biochemical computers with self-reproducing capability.Organized into eight chapters, this book begins with an overview of some theorems dealing with conditions under which universal computation and construction can be exhibited in cellular spaces. This text then presents a design for a machine embedded in a cellular space or a machine that can compute all computable functions and construct a replica of itself in any accessible and sufficiently large region of t

  14. Ethanol production from food waste at high solid contents with vacuum recovery technology

    Science.gov (United States)

    Ethanol production from food wastes does not only solve the environmental issues but also provide renewable biofuel to partially substitute fossil fuels. This study investigated the feasibility of utilization of food wastes for producing ethanol at high solid contents (35%, w/w). Vacuum recovery sys...

  15. Cultural Parallax and Content Analysis: Images of Black Women in High School History Textbooks

    Science.gov (United States)

    Woyshner, Christine; Schocker, Jessica B.

    2015-01-01

    This study investigates the representation of Black women in high school history textbooks. To examine the extent to which Black women are represented visually and to explore how they are portrayed, the authors use a mixed-methods approach that draws on analytical techniques in content analysis and from visual culture studies. Their findings…

  16. Exceptional heat stability of high protein content dispersions containing whey protein particles

    NARCIS (Netherlands)

    Saglam, D.; Venema, P.; Vries, de R.J.; Linden, van der E.

    2014-01-01

    Due to aggregation and/or gelation during thermal treatment, the amount of whey proteins that can be used in the formulation of high protein foods e.g. protein drinks, is limited. The aim of this study was to replace whey proteins with whey protein particles to increase the total protein content and

  17. The content and role of formal contracts in high-tech alliances

    NARCIS (Netherlands)

    Jong, G. de; Klein Woolthuis, R.J.A.

    2009-01-01

    In this study we investigate the governance structure of innovation processes in high-tech alliances, focusing on the content and role of formal contracts. The design of a formal agreement is one of the most important strategic decisions for alliance partners. Drawing upon transaction cost arguments

  18. [Determination of high content of tin in geochemical samples by solid emission spectrometry].

    Science.gov (United States)

    Yao, Jian-Zhen; Hao, Zhi-Hong; Tang, Rui-Ling; Li, Xiao-Jing; Li, Wen-Ge; Zhang, Qin

    2013-11-01

    A method for the determination of high content of tin in geochemical samples by solid emission spectrometry was presented. The dedicated high content tin spectrum standard series was developed. K2S2O7, NaF, Al2O3 and carbon powder were used as buffers and Ge was used as internal standard, and the ratio of sample/matrix/buffer is 1 : 1 : 2. A weak sensitive line (Sn 242. 170 0 nm) was used as the analytical line. The technologies of vertical electrodes, AC arc overlap spectrograph, interception of the exposure, quantitative computer translation spectrum and background correction were used. The determination range is 100-22 350 microg x g(-1), the detection limit is 16.64 microg x g(-1), and the precision is (RSD, n = 12) 4.11%-6.46%. The accuracy of the method has been verified by determination of high content of tin in national geochemical standard samples and the results are in agreement with certified value. The method can be used for measurement directly without dilution of high content of tin in geochemical samples, and it greatly improved the detection upper limit for the determination of tin with solid emission spectroscopy and has certain practical value.

  19. The content and role of formal contracts in high-tech alliances

    NARCIS (Netherlands)

    de Jong, Gjalt; Woolthuis, Rosalinde Ja Klein

    2009-01-01

    In this study we investigate the governance structure of innovation processes in high-tech alliances, focusing on the content and role of formal contracts. The design of a formal agreement is one of the most important strategic decisions for alliance partners. Drawing upon transaction cost arguments

  20. Nanoscale high-content analysis using compositional heterogeneities of single proteoliposomes

    DEFF Research Database (Denmark)

    Mathiasen, Signe; Christensen, Sune M.; Fung, Juan José

    2014-01-01

    heterogeneities can severely skew ensemble-average proteoliposome measurements but also enable ultraminiaturized high-content screens. We took advantage of this screening capability to map the oligomerization energy of the β2-adrenergic receptor using ∼10(9)-fold less protein than conventional assays....

  1. Cultural Parallax and Content Analysis: Images of Black Women in High School History Textbooks

    Science.gov (United States)

    Woyshner, Christine; Schocker, Jessica B.

    2015-01-01

    This study investigates the representation of Black women in high school history textbooks. To examine the extent to which Black women are represented visually and to explore how they are portrayed, the authors use a mixed-methods approach that draws on analytical techniques in content analysis and from visual culture studies. Their findings…

  2. Adhesive performance of washed cottonseed meal at high solid contents and low temperatures

    Science.gov (United States)

    Water-washed cottonseed meal (WCSM) has been shown as a promising biobased wood adhesive. Recently, we prepared WSCM in a pilot scale for promoting its industrial application. In this work, we tested the adhesive strength and viscosity of the adhesive preparation with high solid contents (up to 30%...

  3. Effect of gamma radiation on starch content and cellular morphology of Chlorella pyrenoidosa; Efecto de la radiacion gamma sobre el contenido en almidon y la morfologia celular de Chlorelia pyrenoidosa

    Energy Technology Data Exchange (ETDEWEB)

    Martin, C.; Fernandez, J.

    1983-07-01

    The effect of five doses of gamma radiation (10, 100, 500, 1000 and 5000 Gy) on starch content and fine structure of Chlorella pyrenoidosa has been studied. Results show the accumulation of large amounts of starch on Chlorella cells after irradiation at 500, 1000 and 5000 Gy doses. The starch which is stored in the cytoplasm in form of great grains produces a de organization of the chloroplast structure, observed by electron microscopy techniques. The microphotographs presented also show a significant increase in the cellular size after irradiation up to 500 Gy. (Author) 29 refs.

  4. Cellular transcripts regulated during infections with Highly Pathogenic H5N1 Avian Influenza virus in 3 host systems

    Directory of Open Access Journals (Sweden)

    Noor Suriani M

    2011-04-01

    Full Text Available Abstract Background Highly pathogenic Avian Influenza (HPAI virus is able to infect many hosts and the virus replicates in high levels in the respiratory tract inducing severe lung lesions. The pathogenesis of the disease is actually the outcome of the infection as determined by complex host-virus interactions involving the functional kinetics of large numbers of participating genes. Understanding the genes and proteins involved in host cellular responses are therefore, critical for the elucidation of the mechanisms of infection. Methods Differentially expressed transcripts regulated in a H5N1 infections of whole lung organ of chicken, in-vitro chick embryo lung primary cell culture (CeLu and a continuous Madin Darby Canine Kidney cell line was undertaken. An improved mRNA differential display technique (Gene Fishing™ using annealing control primers that generates reproducible, authentic and long PCR products that are detectable on agarose gels was used for the identification of differentially expressed genes (DEGs. Seven of the genes have been selected for validation using a TaqMan® based real time quantitative PCR assay. Results Thirty seven known and unique differentially expressed genes from lungs of chickens, CeLu and MDCK cells were isolated. Among the genes isolated and identified include heat shock proteins, Cyclin D2, Prenyl (decaprenyl diphosphate synthase, IL-8 and many other unknown genes. The quantitative real time RT-PCR assay data showed that the transcription kinetics of the selected genes were clearly altered during infection by the Highly Pathogenic Avian Influenza virus. Conclusion The Gene Fishing™ technique has allowed for the first time, the isolation and identification of sequences of host cellular genes regulated during H5N1 virus infection. In this limited study, the differentially expressed genes in the three host systems were not identical, thus suggesting that their responses to the H5N1 infection may not share

  5. Evolution of undissolved phases in high-zinc content super-high strength aluminum alloy during ageing

    Institute of Scientific and Technical Information of China (English)

    张坤; 刘志义; 叶呈武; 许晓嫦; 郑青春

    2004-01-01

    The evolution of undissolved phases in the high-zinc content super-high strength aluminum alloy during ageing was investigated by means of SEM and EIS. The results show that undissolved phases of Cu-rich M(AlZnMgCu) exist in the silver-free alloy at solid-solution state. With increasing the ageing time, the precipitation of agehardening precipitates MgZn2 stimulates Zn atoms within the undissolved phases to diffuse into the matrix, and thus the Cu content in the M(AlZnMgCu) phase increases relatively. For the silver-bearing alloy, small addition of Ag promotes the formation of Ag-rich M(A1ZnMgCuAg) undissolved phases and deteriorates mechanical properties of the alloy. At the early stage of ageing, Ag content within the M(AlZnMgCuAg) phases greatly decreases due to rapid diffusing of Ag atoms into the matrix and the co-clustering of Ag and Mg atoms. As the ageing time prolonging, the precipitation of MgZn2 results in the decrease of Zn content in the undissolved phases, and the relative increase of Ag and Mg contents.

  6. High Water Content Material Based on Ba-Bearing Sulphoaluminate Cement

    Institute of Scientific and Technical Information of China (English)

    CHANG Jun; CHENG Xin; LU Lingchao; HUANG Shifeng; YE Zhengmao

    2005-01-01

    A new type of high water content material which is made up of two pastes is prepared, one is made from lime and gypsum, and another is based on Ba-bearing stdphoaluminate cement. It has excellent properties such as slow single paste solidifing,fast double pastes solidifing,fast coagulating and hardening, high early strength, good suspension property at high W/C ratio and low cost. Meanwhile, the properties and hydration mechanism of the material were analyzed by using XRD , DTA- TG and SEM. The hydrated products of new type of high water content material are Ba-bearing ettringite, BaSO4 , aluminum gel and C-S-H gel.

  7. Stimulatory and possible antioxidant effects of High Density Green Photons (HDGP) on cellular systems.

    Science.gov (United States)

    Paslaru, L; Nastase, A; Stefan, L; Florea, R; Sorop, A; Ionescu, E; Popescu, I; Comorasan, S

    2014-01-01

    The interactions between the electromagnetic field and the biological systems were extensively investigated, with remarkable results and advanced technologies. Nevertheless, the visible domain of the spectrum has been rather neglected, since the classic physics did not allow electronic transitions induced by visible light. Recently, the interaction of light with the matter has generated a new scientific domain known in Physics as optical manipulation, with the new concepts of optical matter and optical force. This article presents the results of our work concerning in vitro effects of High Density Green Photons (HDGP) irradiation on cell cultures: stimulation of cell proliferation and migration and a possible antioxidant action.

  8. A galvanic replacement route to prepare strongly fluorescent and highly stable gold nanodots for cellular imaging.

    Science.gov (United States)

    Wang, Chuanxi; Wang, Yu; Xu, Lin; Shi, Xiaodong; Li, Xiangwei; Xu, Xiaowei; Sun, Hongchen; Yang, Bai; Lin, Quan

    2013-02-11

    Fluorescent gold nanodots (GNDs) are an important kind of nanoprobes. Herein, the application of galvanic replacement for the preparation of fluorescent GNDs is reported. Using presynthesized and size-controlled Ag nanodots (Ag NDs) as templates, the as-prepared GNDs have strong fluorescence (quantum yields ~10%) with high stability and surface bioactivity. The resultant GNDs show excellent photoluminescence properties with high photo-, time-, metal-, and pH-stability, which are attributed to the protective surface layer of glutathione (GSH) and the presence of Au(I)-S complexes on the surface of the gold core. GSH, a naturally occurring and readily available tripeptide with carboxyl and amino functional groups, allows good dispersion of the as-prepared GNDs in aqueous solution and favorable biocompatibility. These advantages, combined with their small size, mean that the as-prepared GNDs have potential application in biological labeling, especially as a DNA probe for the specific detection of nucleic acids. In this study, the CAL-27 cells are used as a model to evaluate the fluorescence imaging of GNDs.

  9. Bioenergetics Race: Teaching Cellular Metabolism to High School Students Through an Educational Game

    Directory of Open Access Journals (Sweden)

    I.E.P. Silva

    2011-04-01

    Full Text Available Biochemistry is a science that deals with complex and abstract topics, which are often not understood by high school students, thus interfering in theteaching/learning process. The educational teaching game isa methodology advocated by several educators. The aim of this study was to develop a pedagogical practice to assist in the teaching/learning of high school students. Inthis context we designed a game about Metabolism, which was named "Bioenergetics Race". The game issimple, uses low cost materials and have easy access. The game consists of a board, chips, and a list with 100 multiple questions. Operation of the game: Selected questions lead the students to draw a card and move the corresponding number of squares on the board. The one who reachesthe finish line first is the winner. To consolidate the proposal information, every answer was discussed. At the end of the game the students answered a questionnaire. According to 77.5% of players the level of difficulty was large and 83% rated the game as good for educational proposal. It was found that 100% of students had neverparticipated in such activity in the classroom. It is notable the interest of students andthe abstract and complex topics are better assimilated.

  10. The effect of High Pressure and High Temperature processing on carotenoids and chlorophylls content in some vegetables.

    Science.gov (United States)

    Sánchez, Celia; Baranda, Ana Beatriz; Martínez de Marañón, Iñigo

    2014-11-15

    The effect of High Pressure (HP) and High Pressure High Temperature (HPHT) processing on carotenoid and chlorophyll content of six vegetables was evaluated. In general, carotenoid content was not significantly influenced by HP or HPHT treatments (625 MPa; 5 min; 20, 70 and 117 °C). Regarding chlorophylls, HP treatment caused no degradation or slight increases, while HPHT processes degraded both chlorophylls. Chlorophyll b was more stable than chlorophyll a at 70 °C, but both of them were highly degraded at 117 °C. HPHT treatment at 117 °C provided products with a good retention of carotenoids and colour in the case of red vegetables. Even though the carotenoids also remained in the green vegetables, their chlorophylls and therefore their colour were so affected that milder temperatures need to be applied. As an industrial scale equipment was used, results will be useful for future industrial implementation of this technology.

  11. Simple Cellular Model of Long-Range Multiplicity and $p_t$ Correlations in High-Energy Nuclear Collisions

    CERN Document Server

    Vechernin, V V

    2003-01-01

    A simple cellular model for the description of the long-range multiplicity and $p_t$ correlations in high-energy nuclear collisions originating from the string fusion model is proposed. Three versions of the model: without fusion, with local and with global string fusion are formulated. A Gauss approximation which enables explicit analytical calculations of the correlation functions in some asymptotic cases in the framework of the model is developed. The assumptions of the model and the validity of a Gauss approximation are checked up in the simplest (no fusion) case when the explicit solution of the model can be found. The role of a size of cells is analysed. The modification of the results in the case of non-Poissonian distributions is also discussed.

  12. High-resolution adaptive optics retinal imaging of cellular structure in choroideremia.

    Science.gov (United States)

    Morgan, Jessica I W; Han, Grace; Klinman, Eva; Maguire, William M; Chung, Daniel C; Maguire, Albert M; Bennett, Jean

    2014-09-04

    We characterized retinal structure in patients and carriers of choroideremia using adaptive optics and other high resolution modalities. A total of 57 patients and 18 carriers of choroideremia were imaged using adaptive optics scanning light ophthalmoscopy (AOSLO), optical coherence tomography (OCT), autofluorescence (AF), and scanning light ophthalmoscopy (SLO). Cone density was measured in 59 eyes of 34 patients where the full cone mosaic was observed. The SLO imaging revealed scalloped edges of RPE atrophy and large choroidal vessels. The AF imaging showed hypo-AF in areas of degeneration, while central AF remained present. OCT images showed outer retinal tubulations and thinned RPE/interdigitation layers. The AOSLO imaging revealed the cone mosaic in central relatively intact retina, and cone density was either reduced or normal at 0.5 mm eccentricity. The border of RPE atrophy showed abrupt loss of the cone mosaic at the same location. The AF imaging in comparison with AOSLO showed RPE health may be compromised before cone degeneration. Other disease features, including visualization of choroidal vessels, hyper-reflective clumps of cones, and unique retinal findings, were tabulated to show the frequency of occurrence and model disease progression. The data support the RPE being one primary site of degeneration in patients with choroideremia. Photoreceptors also may degenerate independently. High resolution imaging, particularly AOSLO in combination with OCT, allows single cell analysis of disease in choroideremia. These modalities promise to be useful in monitoring disease progression, and in documenting the efficacy of gene and cell-based therapies for choroideremia and other diseases as these therapies emerge. (ClinicalTrials.gov number, NCT01866371.). Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

  13. High-density lipoprotein maintains skeletal muscle function by modulating cellular respiration in mice.

    Science.gov (United States)

    Lehti, Maarit; Donelan, Elizabeth; Abplanalp, William; Al-Massadi, Omar; Habegger, Kirk M; Weber, Jon; Ress, Chandler; Mansfeld, Johannes; Somvanshi, Sonal; Trivedi, Chitrang; Keuper, Michaela; Ograjsek, Teja; Striese, Cynthia; Cucuruz, Sebastian; Pfluger, Paul T; Krishna, Radhakrishna; Gordon, Scott M; Silva, R A Gangani D; Luquet, Serge; Castel, Julien; Martinez, Sarah; D'Alessio, David; Davidson, W Sean; Hofmann, Susanna M

    2013-11-26

    Abnormal glucose metabolism is a central feature of disorders with increased rates of cardiovascular disease. Low levels of high-density lipoprotein (HDL) are a key predictor for cardiovascular disease. We used genetic mouse models with increased HDL levels (apolipoprotein A-I transgenic [apoA-I tg]) and reduced HDL levels (apoA-I-deficient [apoA-I ko]) to investigate whether HDL modulates mitochondrial bioenergetics in skeletal muscle. ApoA-I ko mice exhibited fasting hyperglycemia and impaired glucose tolerance test compared with wild-type mice. Mitochondria isolated from gastrocnemius muscle of apoA-I ko mice displayed markedly blunted ATP synthesis. Endurance capacity during exercise exhaustion test was impaired in apoA-I ko mice. HDL directly enhanced glucose oxidation by increasing glycolysis and mitochondrial respiration rate in C2C12 muscle cells. ApoA-I tg mice exhibited lower fasting glucose levels, improved glucose tolerance test, increased lactate levels, reduced fat mass, associated with protection against age-induced decline of endurance capacity compared with wild-type mice. Circulating levels of fibroblast growth factor 21, a novel biomarker for mitochondrial respiratory chain deficiencies and inhibitor of white adipose lipolysis, were significantly reduced in apoA-I tg mice. Consistent with an increase in glucose utilization of skeletal muscle, genetically increased HDL and apoA-I levels in mice prevented high-fat diet-induced impairment of glucose homeostasis. In view of impaired mitochondrial function and decreased HDL levels in type 2 diabetes mellitus, our findings indicate that HDL-raising therapies may preserve muscle mitochondrial function and address key aspects of type 2 diabetes mellitus beyond cardiovascular disease.

  14. High School Science Teachers' Perceptions of Teaching Content-Related Reading Comprehension Instruction

    Science.gov (United States)

    Williams, Theresa

    In order to achieve academic success, students must be able to comprehend written material in content-area textbooks. However, a large number of high school students struggle to comprehend science content. Research findings have demonstrated that students make measurable gains in comprehending content-area textbooks when provided quality reading comprehension instruction. The purpose of this study was to gain an understanding of how high school science teachers perceived their responsibility to provide content-related comprehension instruction and 10 high school science teachers were interviewed for this study. Data analysis consisted of open, axial, and selective coding. The findings revealed that 8 out of the 10 participants believed that it is their responsibility to provide reading comprehension. However, the findings also revealed that the participants provided varying levels of reading comprehension instruction as an integral part of their science instruction. The potential for positive social change could be achieved by teachers and administrators. Teachers may use the findings to reflect upon their own personal feelings and beliefs about providing explicit reading comprehension. In addition to teachers' commitment to reading comprehension instruction, administrators could deliberate about professional development opportunities that might improve necessary skills, eventually leading to better comprehension skills for students and success in their education.

  15. Fast repurposing of high-resolution stereo video content for mobile use

    Science.gov (United States)

    Karaoglu, Ali; Lee, Bong Ho; Boev, Atanas; Cheong, Won-Sik; Gotchev, Atanas

    2012-06-01

    3D video content is captured and created mainly in high resolution targeting big cinema or home TV screens. For 3D mobile devices, equipped with small-size auto-stereoscopic displays, such content has to be properly repurposed, preferably in real-time. The repurposing requires not only spatial resizing but also properly maintaining the output stereo disparity, as it should deliver realistic, pleasant and harmless 3D perception. In this paper, we propose an approach to adapt the disparity range of the source video to the comfort disparity zone of the target display. To achieve this, we adapt the scale and the aspect ratio of the source video. We aim at maximizing the disparity range of the retargeted content within the comfort zone, and minimizing the letterboxing of the cropped content. The proposed algorithm consists of five stages. First, we analyse the display profile, which characterises what 3D content can be comfortably observed in the target display. Then, we perform fast disparity analysis of the input stereoscopic content. Instead of returning the dense disparity map, it returns an estimate of the disparity statistics (min, max, meanand variance) per frame. Additionally, we detect scene cuts, where sharp transitions in disparities occur. Based on the estimated input, and desired output disparity ranges, we derive the optimal cropping parameters and scale of the cropping window, which would yield the targeted disparity range and minimize the area of cropped and letterboxed content. Once the rescaling and cropping parameters are known, we perform resampling procedure using spline-based and perceptually optimized resampling (anti-aliasing) kernels, which have also a very efficient computational structure. Perceptual optimization is achieved through adjusting the cut-off frequency of the anti-aliasing filter with the throughput of the target display.

  16. Integrated, High-Throughput, Multiomics Platform Enables Data-Driven Construction of Cellular Responses and Reveals Global Drug Mechanisms of Action.

    Science.gov (United States)

    Norris, Jeremy L; Farrow, Melissa A; Gutierrez, Danielle B; Palmer, Lauren D; Muszynski, Nicole; Sherrod, Stacy D; Pino, James C; Allen, Jamie L; Spraggins, Jeffrey M; Lubbock, Alex L R; Jordan, Ashley; Burns, William; Poland, James C; Romer, Carrie; Manier, M Lisa; Nei, Yuan-Wei; Prentice, Boone M; Rose, Kristie L; Hill, Salisha; Van de Plas, Raf; Tsui, Tina; Braman, Nathaniel M; Keller, M Ray; Rutherford, Stacey A; Lobdell, Nichole; Lopez, Carlos F; Lacy, D Borden; McLean, John A; Wikswo, John P; Skaar, Eric P; Caprioli, Richard M

    2017-03-03

    An understanding of how cells respond to perturbation is essential for biological applications; however, most approaches for profiling cellular response are limited in scope to pre-established targets. Global analysis of molecular mechanism will advance our understanding of the complex networks constituting cellular perturbation and lead to advancements in areas, such as infectious disease pathogenesis, developmental biology, pathophysiology, pharmacology, and toxicology. We have developed a high-throughput multiomics platform for comprehensive, de novo characterization of cellular mechanisms of action. Platform validation using cisplatin as a test compound demonstrates quantification of over 10 000 unique, significant molecular changes in less than 30 days. These data provide excellent coverage of known cisplatin-induced molecular changes and previously unrecognized insights into cisplatin resistance. This proof-of-principle study demonstrates the value of this platform as a resource to understand complex cellular responses in a high-throughput manner.

  17. Selection of hybrids and edible citrus species with a high content in the diosmin functional compound. Modulating effect of plant growth regulators on contents.

    Science.gov (United States)

    Marín, F R; Del Río, J A

    2001-07-01

    The purpose of this study is to identify species, hybrids, and cultivars of edible Citrus species with high contents of diosmin as a functional compound and also to identify the developmental progress of the fruit in which it reaches maximum levels; these findings would be useful for extraction purposes and for the modulating effect of plant growth regulators on diosmin content to increase the level of this flavone. The results obtained reveal that the highest contents of diosmin are present in immature fruits of certain varieties of citron (Buda's finger) and lemon (Meyer), whereas the contents in the edible parts of the fruits are irrelevant from a pharmacological point of view. Similarly, it is shown that it is possible to increase the content of this flavone using hormonal treatments (6-benzylaminopurine and 2,4-dichlorophenoxyacetic acid) during the early stages of fruit growth.

  18. Photoluminescence efficiency of BGaN epitaxial layers with high boron content

    Energy Technology Data Exchange (ETDEWEB)

    Jurkevičius, J.; Mickevičius, J., E-mail: juras.mickevicius@ff.vu.lt; Kadys, A.; Kolenda, M.; Tamulaitis, G.

    2016-07-01

    High-boron-content epitaxial layers of BGaN intended for lattice-matching with AlGaN in UV light emitters were grown on SiC substrate and GaN and AlN templates on sapphire. Photoluminescence (PL) of these layers was studied under quasi-steady-state conditions by varying temperature and excitation intensity. The PL spectra in the samples with different boron content and their dynamics evidence formation of boron-rich regions occupying a small fraction of the total layer volume and acting as the emission killers. The room-temperature PL efficiency of the BGaN epilayers was estimated and shown to drastically decrease at increasing boron content with no significant correlation with either the type of substrate/template or technological conditions of the layer deposition.

  19. Determination of Ge content in high concentration Ge-doped Czochralski Si single crystals by FTIR

    Institute of Scientific and Technical Information of China (English)

    JIANG Zhongwei; ZHANG Weilian; NIU Xinhuan

    2005-01-01

    SiGe single crystals with different Ge concentrations were measured by Fourier transform infrared (FTIR) spectroscopy at room temperature (RT) and 10 K. A new peak appears at the wave number of 710 cm-1 and the spectroscopy becomes clearer with an increase in Ge content. The absorption strength and wave sharp of the 710 cm-1 peak are independent of temperature. The relation of the absorption coefficient amax, the band width of half maximum (BWHM) W1/2 of the 710 cm-1 peak, and the Ge concentration is determined with the Ge content obtained by SEM-EDX. The conversion factor is k = 1.211 at 10 K. Therefore, the Ge content in high concentration Ge doped CZ-Si single crystals can be determined by FTIR.

  20. Structure and Content Analysis for Vocational High School Website in Indonesia

    Science.gov (United States)

    Subagja, H.; Abdullah, A. G.; Trisno, B.; Nandiyanto, A. B. D.

    2017-03-01

    Statistics about the condition of the school’s website in Indonesia is still difficult. This study aims to determine website quality in terms of completeness of content’s criteria of Vocational High School (VHS) in West Java, Indonesia. The method used is the content analysis and survey. Content analysis is reviewing the documents comprising the general category, while the survey is a observation process to get the facts from 272 school websites. Aspects of the structure and content of school website are including institutional information, educators and education personnel, curriculum, student, infrastructure, school achievement, and public access. The results of this study showed the average quality of the VHS website in West Java is still low. The recommendations are needed to improve the quality of the school website.

  1. Deposition of High Conductivity Low Silver Content Materials by Screen Printing

    Directory of Open Access Journals (Sweden)

    Eifion Jewell

    2015-05-01

    Full Text Available A comprehensive experimental investigation has been carried out into the role of film thickness variation and silver material formulation on printing capability in the screen printing process. A full factorial experiment was carried out where two formulations of silver materials were printed through a range of screens to a polyester substrate under a set of standard conditions. The materials represented a novel low silver content (45%–49% polymer material and traditional high silver content (65%–69% paste. The resultant prints were characterised topologically and electrically. The study shows that more cost effective use of the silver in the ink was obtained with the low silver polymer materials, but that the electrical performance was more strongly affected by the mesh being used (and hence film thickness. Thus, while optimum silver use could be obtained using materials with a lower silver content, this came with the consequence of reduced process robustness.

  2. A High Resolution Capacitive Sensing System for the Measurement of Water Content in Crude Oil

    Directory of Open Access Journals (Sweden)

    Muhammad Zubair Aslam

    2014-06-01

    Full Text Available This paper presents the design of a non-intrusive system to measure ultra-low water content in crude oil. The system is based on a capacitance to phase angle conversion method. Water content is measured with a capacitance sensor comprising two semi-cylindrical electrodes mounted on the outer side of a glass tube. The presence of water induces a capacitance change that in turn converts into a phase angle, with respect to a main oscillator. A differential sensing technique is adopted not only to ensure high immunity against temperature variation and background noise, but also to eliminate phase jitter and amplitude variation of the main oscillator that could destabilize the output. The complete capacitive sensing system was implemented in hardware and experiment results using crude oil samples demonstrated that a resolution of ±50 ppm of water content in crude oil was achieved by the proposed design.

  3. Reduction of phorbol ester content in jatropha cake using high energy gamma radiation

    Directory of Open Access Journals (Sweden)

    Runumi Gogoi

    2014-07-01

    Full Text Available In this paper, an attempt has been made to remove phorbol ester present in jatropha cake by exposing it to gamma radiation. A sensitizer was also used to accelerate the degradation of phorbol esters. The phorbol ester content in the cake was estimated by high performance liquid chromatography (HPLC. It was observed that gamma irradiation of the jatropha seed cake was effective in reducing the phorbol ester content. Originally, the phorbol ester content in the cake was found as 0.29 mg/g, which on exposure to radiation was reduced by 33.4% and 96% with radiation dose of 30 and 125 kGy respectively. The presence of a sensitizer was found to enhances the susceptibility of phorbol esters degradation by oxidative degradation on exposure to ionizing radiation.

  4. In vitro testing of cellular response to ultra high frequency electromagnetic field radiation.

    Science.gov (United States)

    Pavicic, Ivan; Trosic, Ivancica

    2008-08-01

    The aim of this study was to evaluate whether low-level, ultra high frequency (UHF) irradiation of 935 MHz influences the cell structure and growth of V79 cells. UHF field was generated inside a Gigahertz Transversal Electromagnetic Mode cell (GTEM-cell) with a Hewlett-Packard signal generator. The electric field strength was 8.2+/-0.3 V/cm and the average specific absorption rate (SAR) was calculated to be 0.12 W/kg. Cell samples were cultivated in a humidified atmosphere at 37 degrees C with 5% CO2. Prepared cell samples were exposed to a 935 MHz continuous wave frequency field for 1, 2, and 3 h. The structure of microtubule proteins has been determined using the immunocytochemical method. Cell growth was determined by cell counts for each hour of exposure during five post-exposure days. Negative- and positive-cell controls were included into the experimental procedure. In comparison with control cells, the microtubule structure clearly altered after 3h of irradiation (pgrowth was noted in cells exposed for 3h three days after irradiation (pcell growth.

  5. High-Reliability Low-Ag-Content Sn-Ag-Cu Solder Joints for Electronics Applications

    Science.gov (United States)

    Shnawah, Dhafer Abdulameer; Said, Suhana Binti Mohd; Sabri, Mohd Faizul Mohd; Badruddin, Irfan Anjum; Che, Fa Xing

    2012-09-01

    Sn-Ag-Cu (SAC) alloy is currently recognized as the standard lead-free solder alloy for packaging of interconnects in the electronics industry, and high- Ag-content SAC alloys are the most popular choice. However, this choice has been encumbered by the fragility of the solder joints that has been observed in drop testing as well as the high cost of the Ag itself. Therefore, low-Ag-content SAC alloy was considered as a solution for both issues. However, this approach may compromise the thermal-cycling performance of the solders. Therefore, to enhance the thermal-cycling reliability of low-Ag-content SAC alloys without sacrificing their drop-impact performance, alloying elements such as Mn, Ce, Ti, Bi, In, Sb, Ni, Zn, Al, Fe, and Co were selected as additions to these alloys. However, research reports related to these modified SAC alloys are limited. To address this paucity, the present study reviews the effect of these minor alloying elements on the solder joint reliability of low-Ag-content SAC alloys in terms of thermal cycling and drop impact. Addition of Mn, Ce, Bi, and Ni to low-Ag-content SAC solder effectively improves the thermal-cycling reliability of joints without sacrificing the drop-impact performance. Taking into consideration the improvement in the bulk alloy microstructure and mechanical properties, wetting properties, and growth suppression of the interface intermetallic compound (IMC) layers, addition of Ti, In, Sb, Zn, Al, Fe, and Co to low-Ag-content SAC solder has the potential to improve the thermal-cycling reliability of joints without sacrificing the drop-impact performance. Consequently, further investigations of both thermal-cycling and drop reliability of these modified solder joints must be carried out in future work.

  6. S-RHT FIXED-BED HYDROTREATING TECHNOLOGY FOR RESIDUE WITH HIGH SULFUR CONTENT

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    To meet the demands of processing residue with high sulfur content, Fushun Research Institute of Petroleum and Petrochemicals (FRIPP) successfully developed the fixed-bed residue hydrotreating technology (S-RHT), which is suitable for treating high sulfur residue to produce diesel oil with low sulfur content. the hydrotreated 350 ℃+ residue is also a good feedstock for RFCC. Based on this technology, Maoming Petrochemical Company constructed the hydrotreating unit with a capacity of 2 Mt/a to treat high sulfur residue, in which loaded FZC series catalysts were developed by FRIPP. The unit was first put into commercial operation at the end of 1999. The commercial operation showed that the catalysts have good activities and the products meet the designed specifications.

  7. Effect of Lower and Upper Body High Intensity Training on Genes Associated with Cellular Stress Response

    Science.gov (United States)

    Kochanowicz, Andrzej; Kochanowicz, Kazimierz; Mieszkowski, Jan; Niespodziński, Bartłomiej; Sawczyn, Stanisław

    2017-01-01

    This study aimed to compare the effect of upper and lower body high intensity exercise (HIE) on select gene expression in athletes. Fourteen elite male artistic gymnasts (age 20.9 ± 2.6 years; weight 68.6 ± 7.2 kg; fat free mass 63.6 ± 6.7 kg; height 1.70 ± 0.04 m) performed lower and upper body 30 s Wingate Tests (WAnTs) before and after eight weeks of specific HIIT. Two milliliters of blood was collected before and after (5, 30 min, resp.) lower and upper body WAnTs, and select gene expression was determined by PCR. Eight weeks of HIIT caused a significant increase in maximal power (722 to 751 Wat), relative peak power in the lower body WAnTs (10.1 to 11 W/kg), mean power (444 to 464 W), and relative mean power (6.5 to 6.8 W/kg). No significant differences in lower versus upper body gene expression were detected after HIIT, and a significant decrease in the IL6/IL10 ratio was observed after lower (−2∧0.57 p = 0.0019) and upper (−2∧0.5 p = 0.03) WAnTs following eight weeks of HIIT. It is hypothesized that a similar adaptive response to exercise may be obtained by lower and upper body exercise. PMID:28589135

  8. High-Aspect Ratio Bio-Metallic Nanocomposites for Cellular Interactions

    Science.gov (United States)

    Deodhar, Sneha; Huckaby, Justin; Delahoussaye, Miles; DeCoster, Mark A.

    2014-08-01

    We synthesized high aspect ratio composites with biological and metal components. Scanning electron microscopy (SEM) and Transmission Electron Microscopy (TEM) revealed linear morphology and smooth surface texture. SEM, TEM and light microscopy showed that composites have scalable dimensions from nano- to micro-, with diameters as low as 60 nm, lengths exceeding 150 pm, and average aspect ratio of 100. The structures are stable, remaining intact for over one year in dried form and in liquid, and did not aggregate, in contrast to metal nanoparticles such as iron and copper. Many metal nanoparticles are toxic to cells, limiting their use for biological applications. The bio-metallic composites characterized here showed lower toxicity compared to their precursor metal nanoparticles in brain tumor cell cultures. Due to these more biocompatible properties, we tested the ability of the composites to interact with cells. Zeta potential analysis indicated that composites carry a net negative charge (-24.3 ± 2.2 mV), while the starting metal nanoparticles measured (43.3 ± 2.4 mV). We labeled the composites with poly-l-lysine fluorescein isothiocyanate (PLL-FITC), which shifted the potential to 3.5 ± 2.9 mV. It was observed by fluorescence microscopy that composites smaller than cells were internalized by some cells and larger composites remained outside. Cells became fluorescent over time due to leakage of PLL-FITC from the composites which lost fluorescence over time. Higher biocompatibility, low aggregation, and ability to control size distribution of the linear composites may make them ideal vehicles to deliver drugs or other materials to cells, and may be used as a scaffolding material for cells.

  9. Effect of religious context on the content of visual hallucinations in individuals high in religiosity.

    Science.gov (United States)

    Reed, Phil; Clarke, Natasha

    2014-03-30

    This study investigated the interaction between the current environment and personality factors associated with religiosity in determining the content of false perceptions (used as a model for hallucinations). A primed word-detection task was used to investigate the effect of a 'religious' context on false perceptions in individuals scoring highly on religiosity. After a subliminal prime, participants viewed letter strings, and stated any words that they saw. The prime and the actual words could have a religious connotation or not. Participants measuring high on religiosity were more likely to report false perceptions of a religious type than participants low on religiosity. It is suggested that context affects the content of false perceptions through the activation of stored beliefs and values, which vary between individuals, offering a mechanism for the effect of context on idiosyncratic content of hallucinations in schizophrenia. The effect of context and individual differences on false-perception content in the current study provides possibilities for future work regarding the underlying nature of hallucinations and their treatment.

  10. Effect of droplet heat content distribution on humping formation in high speed GMAW

    Institute of Scientific and Technical Information of China (English)

    Ji CHEN; Chuansong WU

    2011-01-01

    The momentum of strong backward flowing melt jet and the thermal action from transferred droplets are two dominating factors affecting the formation of humping bead in high speed gas metal arc welding (GMAW).Appropriate describing the influence of the distribution mode of droplet heat content in the weld pool is essential to understand the physical mechanism of humping bead formation.Based on the experimental results,four kinds of droplet heat content distribution modes are proposed and employed to calculate the transient evolution of the temperature field and weld pool during high speed GMAW process.Through making comparison of predicted and measured weld bead dimensions,a suitable and adaptive distribution mode of droplet heat content is found,i.e.,droplet heat content is distributed in bottom layer of gouging region at the front of weld pool,and is averagely distributed in the whole layer at the rear of weld pool.The proposed mode is also validated by experimental observation of the weld pool images and measured by geometric dimensions of the weld bead.

  11. Influence of Ta Content in High Purity Niobium on Cavity Performance Preliminary Results*

    CERN Document Server

    Kneisel, P

    2004-01-01

    In a previous paper* a program designed to study the influence of the residual tantalum content on the superconducting properties of pure niobium metal for RF cavities was outlined. The main rationale for this program was based on a potential cost reduction for high purity niobium, if a less strict limit on the chemical specification for Ta content, which is not significantly affecting the RRR–value, could be tolerated for high performance cavities. Four ingots with different Ta contents have been melted and transformed into sheets. In each manufacturing step the quality of the material has been monitored by employing chemical analysis, neutron activation analysis, thermal conductivity measurements and evaluation of the mechanical properties. The niobium sheets have been scanned for defects by an eddy current device. From three of the four ingots—Ta contents 100, 600 and 1,200 wppm—two single cell cavities each of the CEBAF variety have been fabricated and a series of tests on each ...

  12. A Categorical Content Analysis of Highly Cited Literature Related to Trends and Issues in Special Education.

    Science.gov (United States)

    Arden, Sarah V; Pentimonti, Jill M; Cooray, Rochana; Jackson, Stephanie

    2017-07-01

    This investigation employs categorical content analysis processes as a mechanism to examine trends and issues in a sampling of highly cited (100+) literature in special education journals. The authors had two goals: (a) broadly identifying trends across publication type, content area, and methodology and (b) specifically identifying articles with disaggregated outcomes for students with learning disabilities (LD). Content analyses were conducted across highly cited (100+) articles published during a 20-year period (1992-2013) in a sample ( n = 3) of journals focused primarily on LD, and in one broad, cross-categorical journal recognized for its impact in the field. Results indicated trends in the article type (i.e., commentary and position papers), content (i.e., reading and behavior), and methodology (i.e., small proportions of experimental and quasi-experimental designs). Results also revealed stability in the proportion of intervention research studies when compared to previous analyses and a decline in the proportion of those that disaggregated data specifically for students with LD.

  13. Multiparametric High Content Analysis for assessment of neurotoxicity in differentiated neuronal cell lines and human embryonic stem cell-derived neurons.

    Science.gov (United States)

    Wilson, Melinda S; Graham, James R; Ball, Andrew J

    2014-05-01

    The potential for adverse neurotoxic reactions in response to therapeutics and environmental hazards continues to prompt development of novel cell-based assays to determine neurotoxic risk. A challenge remains to characterize and understand differences between assays and between neuronal cellular models in their responses to neurotoxicants if scientists are to determine the optimal model, or combination of models, for neurotoxicity screening. Most studies to date have focused on developmental neurotoxicity applications. This study reports the development of a robust multiparameter High Content Analysis (HCA) assay for neurotoxicity screening in three differentiated neuronal cell models - SH-SY5Y, PC12 and human embryonic stem cell-derived hN2™ cells. Using a multiplexed detection reagent panel (Hoechst nuclear stain; antibodies against βIII-Tubulin and phosphorylated neurofilament subunit H, and Mitotracker(®) Red CMXRos), a multiparametric HCA assay was developed and used to characterize a test set of 36 chemicals. HCA data generated were compared to data generated using MTT and LDH assays under the same assay conditions. Data showed that multiparametric High Content Analysis of differentiated neuronal cells is feasible, and represents a highly effective method for obtaining large quantities of robust data on the neurotoxic effects of compounds compared with cytotoxicity assays like MTT and LDH. Significant differences were observed between the responses to compounds across the three cellular models tested, illustrating the heterogeneity in responses to neurotoxicants across different cell types. This study provides data strongly supporting the use of cellular imaging as a tool for neurotoxicity assessment in differentiated neuronal cells, and provides novel insights into the neurotoxic effects of a test set of compounds upon differentiated neuronal cell lines and human embryonic stem cell-derived neurons. Copyright © 2014 Elsevier Inc. All rights reserved.

  14. High Content Analysis Provides Mechanistic Insights on the Pathways of Toxicity Induced by Amine-Modified Polystyrene Nanoparticles

    Science.gov (United States)

    Anguissola, Sergio; Garry, David; Salvati, Anna; O'Brien, Peter J.; Dawson, Kenneth A.

    2014-01-01

    The fast-paced development of nanotechnology needs the support of effective safety testing. We have developed a screening platform measuring simultaneously several cellular parameters for exposure to various concentrations of nanoparticles (NPs). Cell lines representative of different organ cell types, including lung, endothelium, liver, kidney, macrophages, glia, and neuronal cells were exposed to 50 nm amine-modified polystyrene (PS-NH2) NPs previously reported to induce apoptosis and to 50 nm sulphonated and carboxyl-modified polystyrene NPs that were reported to be silent. All cell lines apart from Raw 264.7 executed apoptosis in response to PS-NH2 NPs, showing specific sequences of EC50 thresholds; lysosomal acidification was the most sensitive parameter. Loss of mitochondrial membrane potential and plasma membrane integrity measured by High Content Analysis resulted comparably sensitive to the equivalent OECD-recommended assays, allowing increased output. Analysis of the acidic compartments revealed good cerrelation between size/fluorescence intensity and dose of PS-NH2 NPs applied; moreover steatosis and phospholipidosis were observed, consistent with the lysosomal alterations revealed by Lysotracker green; similar responses were observed when comparing astrocytoma cells with primary astrocytes. We have established a platform providing mechanistic insights on the response to exposure to nanoparticles. Such platform holds great potential for in vitro screening of nanomaterials in highthroughput format. PMID:25238162

  15. Adaptation of a Cell-Based High Content Screening System for the In-Depth Analysis of Celiac Biopsy Tissue.

    Science.gov (United States)

    Cooper, Sarah E J; Mohamed, Bashir M; Elliott, Louise; Davies, Anthony Mitchell; Feighery, Conleth F; Kelly, Jacinta; Dunne, Jean

    2015-01-01

    The IN Cell Analyzer 1000 possesses several distinguishing features that make it a valuable tool in research today. This fully automated high content screening (HCS) system introduced quantitative fluorescent microscopy with computerized image analysis for use in cell-based analysis. Previous studies have focused on live cell assays, where it has proven to be a powerful and robust method capable of providing reproducible, quantitative data. Using HCS as a tool to investigate antigen expression in duodenal biopsies, we developed a novel approach to tissue positioning and mapping. We adapted IN Cell Analyzer 1000's image acquisition and analysis software for the investigation of tissue transglutaminase (tTG) and smooth muscle alpha-actin (SM α-actin) staining in paraffin-embedded duodenal tissue sections from celiac patients and healthy controls. These innovations allowed a quantitative analysis of cellular structure and protein expression. The results from routine biopsy material indicated the intensity of protein expression was altered in celiac disease compared to normal biopsy material.

  16. A Novel Automated High-Content Analysis Workflow Capturing Cell Population Dynamics from Induced Pluripotent Stem Cell Live Imaging Data

    Science.gov (United States)

    Kerz, Maximilian; Folarin, Amos; Meleckyte, Ruta; Watt, Fiona M.; Dobson, Richard J.; Danovi, Davide

    2016-01-01

    Most image analysis pipelines rely on multiple channels per image with subcellular reference points for cell segmentation. Single-channel phase-contrast images are often problematic, especially for cells with unfavorable morphology, such as induced pluripotent stem cells (iPSCs). Live imaging poses a further challenge, because of the introduction of the dimension of time. Evaluations cannot be easily integrated with other biological data sets including analysis of endpoint images. Here, we present a workflow that incorporates a novel CellProfiler-based image analysis pipeline enabling segmentation of single-channel images with a robust R-based software solution to reduce the dimension of time to a single data point. These two packages combined allow robust segmentation of iPSCs solely on phase-contrast single-channel images and enable live imaging data to be easily integrated to endpoint data sets while retaining the dynamics of cellular responses. The described workflow facilitates characterization of the response of live-imaged iPSCs to external stimuli and definition of cell line–specific, phenotypic signatures. We present an efficient tool set for automated high-content analysis suitable for cells with challenging morphology. This approach has potentially widespread applications for human pluripotent stem cells and other cell types. PMID:27256155

  17. High content analysis provides mechanistic insights on the pathways of toxicity induced by amine-modified polystyrene nanoparticles.

    Science.gov (United States)

    Anguissola, Sergio; Garry, David; Salvati, Anna; O'Brien, Peter J; Dawson, Kenneth A

    2014-01-01

    The fast-paced development of nanotechnology needs the support of effective safety testing. We have developed a screening platform measuring simultaneously several cellular parameters for exposure to various concentrations of nanoparticles (NPs). Cell lines representative of different organ cell types, including lung, endothelium, liver, kidney, macrophages, glia, and neuronal cells were exposed to 50 nm amine-modified polystyrene (PS-NH2) NPs previously reported to induce apoptosis and to 50 nm sulphonated and carboxyl-modified polystyrene NPs that were reported to be silent. All cell lines apart from Raw 264.7 executed apoptosis in response to PS-NH2 NPs, showing specific sequences of EC50 thresholds; lysosomal acidification was the most sensitive parameter. Loss of mitochondrial membrane potential and plasma membrane integrity measured by High Content Analysis resulted comparably sensitive to the equivalent OECD-recommended assays, allowing increased output. Analysis of the acidic compartments revealed good cerrelation between size/fluorescence intensity and dose of PS-NH2 NPs applied; moreover steatosis and phospholipidosis were observed, consistent with the lysosomal alterations revealed by Lysotracker green; similar responses were observed when comparing astrocytoma cells with primary astrocytes. We have established a platform providing mechanistic insights on the response to exposure to nanoparticles. Such platform holds great potential for in vitro screening of nanomaterials in highthroughput format.

  18. Optimizing Cationic and Neutral Lipids for Efficient Gene Delivery at High Serum Content

    Science.gov (United States)

    Majzoub, Ramsey N.; Hwu, Yeu-kuang; Liang, Keng S.; Leal, Cecília; Safinya, Cyrus R.

    2014-01-01

    Background Cationic liposome (CL)-DNA complexes are promising gene delivery vectors with potential applications in gene therapy. A key challenge in creating CL-DNA complexes for applications is that their transfection efficiency (TE) is adversely affected by serum. In particular, little is known about the effects of high serum contents on TE even though this may provide design guidelines for applications in vivo. Methods We prepared CL-DNA complexes in which we varied the neutral lipid (DOPC, glycerol-monooleate (GMO), cholesterol), the headgroup charge and chemical structure of the cationic lipid, and the ratio of neutral to cationic lipid; we then measured the TE of these complexes as a function of serum content and assessed their cytotoxicity. We tested selected formulations in two human cancer cell lines (M21/melanoma and PC-3/prostate cancer). Results In the absence of serum, all CL-DNA complexes of custom-synthesized multivalent lipids show high TE. Certain combinations of multivalent lipids and neutral lipids, such as MVL5(5+)/GMO-DNA complexes or complexes based on the dendritic-headgroup lipid TMVLG3(8+) exhibited high TE both in the absence and presence of serum. Although their TE still dropped to a small extent in the presence of serum, it reached or surpassed that of benchmark commercial transfection reagents, in particular at high serum content. Conclusions Two-component vectors (one multivalent cationic lipid and one neutral lipid) can rival or surpass benchmark reagents at low and high serum contents (up to 50%, v/v). We suggest guidelines for optimizing the serum resistance of CL-DNA complexes based on a given cationic lipid. PMID:24753287

  19. Active Learning Strategies for Phenotypic Profiling of High-Content Screens.

    Science.gov (United States)

    Smith, Kevin; Horvath, Peter

    2014-06-01

    High-content screening is a powerful method to discover new drugs and carry out basic biological research. Increasingly, high-content screens have come to rely on supervised machine learning (SML) to perform automatic phenotypic classification as an essential step of the analysis. However, this comes at a cost, namely, the labeled examples required to train the predictive model. Classification performance increases with the number of labeled examples, and because labeling examples demands time from an expert, the training process represents a significant time investment. Active learning strategies attempt to overcome this bottleneck by presenting the most relevant examples to the annotator, thereby achieving high accuracy while minimizing the cost of obtaining labeled data. In this article, we investigate the impact of active learning on single-cell-based phenotype recognition, using data from three large-scale RNA interference high-content screens representing diverse phenotypic profiling problems. We consider several combinations of active learning strategies and popular SML methods. Our results show that active learning significantly reduces the time cost and can be used to reveal the same phenotypic targets identified using SML. We also identify combinations of active learning strategies and SML methods which perform better than others on the phenotypic profiling problems we studied.

  20. Biocompatible Ni-free Zr-based bulk metallic glasses with high-Zr-content: compositional optimization for potential biomedical applications.

    Science.gov (United States)

    Hua, Nengbin; Huang, Lu; Chen, Wenzhe; He, Wei; Zhang, Tao

    2014-11-01

    The present study designs and prepares Ni-free Zr60+xTi2.5Al10Fe12.5-xCu10Ag5 (at.%, x=0, 2.5, 5) bulk metallic glasses (BMGs) by copper mold casting for potential biomedical application. The effects of Zr content on the in vitro biocompatibility of the Zr-based BMGs are evaluated by investigating mechanical properties, bio-corrosion behavior, and cellular responses. It is found that increasing the content of Zr is favorable for the mechanical compatibility with a combination of low Young's modulus, large plasticity, and high notch toughness. Electrochemical measurements demonstrate that the Zr-based BMGs are corrosion resistant in a phosphate buffered saline solution. The bio-corrosion resistance of BMGs is improved with the increase in Zr content, which is attributed to the enrichment in Zr and decrease in Al concentration in the surface passive film of alloys. Regular cell responses of mouse MC3T3-E1 cells, including cell adhesion and proliferation, are observed on the Zr-Ti-Al-Fe-Cu-Ag BMGs, which reveals their general biosafety. The high-Zr-based BMGs exhibit a higher cell proliferation activity in comparison with that of pure Zr and Ti-6Al-4V alloy. The effects of Zr content on the in vitro biocompatibility can be used to guide the future design of biocompatible Zr-based BMGs. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Variations in pore characteristics in high volatile bituminous coals: Implications for coal bed gas content

    Science.gov (United States)

    Mastalerz, Maria; Drobniak, A.; Strapoc, D.; Solano-Acosta, W.; Rupp, J.

    2008-01-01

    The Seelyville Coal Member of the Linton Formation (Pennsylvanian) in Indiana was studied to: 1) understand variations in pore characteristics within a coal seam at a single location and compare these variations with changes occurring between the same coal at different locations, 2) elaborate on the influence of mineral-matter and maceral composition on mesopore and micropore characteristics, and 3) discuss implications of these variations for coal bed gas content. The coal is high volatile bituminous rank with R0 ranging from 0.57% to 0.60%. BET specific surface areas (determined by nitrogen adsorption) of the coals samples studied range from 1.8 to 22.9??m2/g, BJH adsorption mesopore volumes from 0.0041 to 0.0339??cm3/g, and micropore volumes (determined by carbon dioxide adsorption) from 0.0315 to 0.0540??cm3/g. The coals that had the largest specific surface areas and largest mesopore volumes occur at the shallowest depths, whereas the smallest values for these two parameters occur in the deepest coals. Micropore volumes, in contrast, are not depth-dependent. In the coal samples examined for this study, mineral-matter content influenced both specific surface area as well as mesopore and micropore volumes. It is especially clear in the case of micropores, where an increase in mineral-matter content parallels the decrease of micropore volume of the coal. No obvious relationships were observed between the total vitrinite content and pore characteristics but, after splitting vitrinite into individual macerals, we see that collotelinite influences both meso- and micropore volume positively, whereas collodetrinite contributes to the reduction of mesopore and micropore volumes. There are large variations in gas content within a single coal at a single location. Because of this variability, the entire thickness of the coal must be desorbed in order to determine gas content reliably and to accurately calculate the level of gas saturation. ?? 2008 Elsevier B.V. All

  2. Non-isothermal Crystallization Kinetics of Spinels in Vanadium Slag with High CaO Content

    Science.gov (United States)

    Zhou, Wang; Xie, Bing; Tan, Wen-Feng; Diao, Jiang; Zhang, Xie; Li, Hong-Yi

    2016-09-01

    This paper investigated the non-isothermal crystallization kinetics of the spinel crystals in vanadium slags containing high CaO content. Experiments were performed in combination with theoretical calculation to address this issue, and statistical analyses based on the Crystal Size Distribution theory. The results indicate that low cooling rate and high CaO content benefit the growth of spinel crystals. The growth mechanism is revealed to be controlled by interface reactions and diffusion at the cooling rates of 5 K/min and 15 K/min, respectively. However, at higher temperatures (>1673 K), the growth of spinel crystals is controlled by nucleation. While the temperature is decreased to 1523 K at the cooling rate of 5 K/min, the mean diameter of spinel crystals could reach 36.44 μm. Experimental results combining with theoretical reveal that low cooling rate benefits spinels growth, especially for the interval of 1523 K-1200 K.

  3. Nonisothermal Thermogravimetric Analysis of Thai Lignite with High CaO Content

    Directory of Open Access Journals (Sweden)

    Pakamon Pintana

    2013-01-01

    Full Text Available Thermal behaviors and combustion kinetics of Thai lignite with different SO3-free CaO contents were investigated. Nonisothermal thermogravimetric method was carried out under oxygen environment at heating rates of 10, 30, and 50°C min−1 from ambient up to 1300°C. Flynn-Wall-Ozawa (FWO and Kissinger-Akahira-Sunose (KAS methods were adopted to estimate the apparent activation energy (E for the thermal decomposition of these coals. Different thermal degradation behaviors were observed in lignites with low (14% and high (42% CaO content. Activation energy of the lignite combustion was found to vary with the conversion fraction. In comparison with the KAS method, higher E values were obtained by the FWO method for all conversions considered. High CaO lignite was observed to have higher activation energy than the low CaO coal.

  4. Aquatic heterotrophic bacteria have highly flexible phosphorus content and biomass stoichiometry.

    Science.gov (United States)

    Godwin, Casey M; Cotner, James B

    2015-10-01

    Bacteria are central to the cycling of carbon (C), nitrogen (N) and phosphorus (P) in every ecosystem, yet our understanding of how tightly these cycles are coupled to bacterial biomass composition is based upon data from only a few species. Bacteria are commonly assumed to have high P content, low biomass C:P and N:P ratios, and inflexible stoichiometry. Here, we show that bacterial assemblages from lakes exhibit unprecedented flexibility in their P content (3% to less than 0.01% of dry mass) and stoichiometry (C:N:P of 28: 7: 1 to more than 8500: 1200: 1). The flexibility in C:P and N:P stoichiometry was greater than any species or assemblage, including terrestrial and aquatic autotrophs, and suggests a highly dynamic role for bacteria in coupling multiple element cycles.

  5. Effect of spinel content on the properties of phosphoric acid bonded high alumina castables

    Institute of Scientific and Technical Information of China (English)

    Zichun Yang; Hongwei Duan; Lin Li; Shuqin Li; Wen Ni

    2003-01-01

    In order to study the effect of fused spinel on the properties of phosphoric acid bonded high alumina castables, samples with different contents of fused spinel were prepared. The results show that when the contents of the fused spinel are between 8% and 16% (mass fraction), the castables have good properties. The castables overcome the shortages of the phosphoric acid bonded high alumina castables with bauxite cement as a hardening promoter. The experiments demonstrate that most of the service properties of the castables with fused spinel are better than those of the normal phosphoric acid bonded castables which use bauxite cement as a hardening promoter. The examination of the materials indicates that free MgO inclusions in the spinel powder can promote the hardening of the castables.

  6. Electrodeposition of Sm–Co film with high Sm content from aqueous solution

    Energy Technology Data Exchange (ETDEWEB)

    Long, Xiong-fei; Guo, Guang-hua, E-mail: guogh@mail.csu.edu.cn; Li, Xin-hua; Xia, Qing-lin; Zhang, Jin-fang

    2013-12-02

    Sm–Co films with high Sm content were electrodeposited by potentiostat technique from the aqueous solution containing glycine as a complexing agent. The Cyclic Voltammetry measurements showed that the glycine plays a crucial role in the electrodeposition of Sm–Co films. The forming of stable complex [Co{sup II}Sm{sup III}(Gly{sup −}){sub 2}(HGly{sup ±})]{sup 3+} facilitated the codeposition of Sm with Co at relatively low potential, which made it possible to fabricate the Sm–Co films with high Sm content. The Sm–Co film with 43 at.% Sm had been obtained. X-ray diffraction showed that the as-deposited Sm–Co films were amorphous. After annealing at 600 °C for 2 h in Ar atmosphere, the films became crystalline and the permanent Sm{sub 2}Co{sub 17} phase was formed, which significantly enhanced the coercivity of the films. - Highlights: • The Sm–Co film with high Sm content is electrodeposited from aqueous solutions. • The addition of glycine facilitates the codeposition of Sm with Co at relatively low potential. • The Sm{sub 2}Co{sub 17} phase is formed in the annealed Sm–Co film.

  7. High-Content Microscopy Analysis of Subcellular Structures: Assay Development and Application to Focal Adhesion Quantification.

    Science.gov (United States)

    Kroll, Torsten; Schmidt, David; Schwanitz, Georg; Ahmad, Mubashir; Hamann, Jana; Schlosser, Corinne; Lin, Yu-Chieh; Böhm, Konrad J; Tuckermann, Jan; Ploubidou, Aspasia

    2016-07-01

    High-content analysis (HCA) converts raw light microscopy images to quantitative data through the automated extraction, multiparametric analysis, and classification of the relevant information content. Combined with automated high-throughput image acquisition, HCA applied to the screening of chemicals or RNAi-reagents is termed high-content screening (HCS). Its power in quantifying cell phenotypes makes HCA applicable also to routine microscopy. However, developing effective HCA and bioinformatic analysis pipelines for acquisition of biologically meaningful data in HCS is challenging. Here, the step-by-step development of an HCA assay protocol and an HCS bioinformatics analysis pipeline are described. The protocol's power is demonstrated by application to focal adhesion (FA) detection, quantitative analysis of multiple FA features, and functional annotation of signaling pathways regulating FA size, using primary data of a published RNAi screen. The assay and the underlying strategy are aimed at researchers performing microscopy-based quantitative analysis of subcellular features, on a small scale or in large HCS experiments. © 2016 by John Wiley & Sons, Inc.

  8. Retrogradation of Maize Starch after High Hydrostatic Pressure Gelation: Effect of Amylose Content and Depressurization Rate

    KAUST Repository

    Yang, Zhi

    2016-05-24

    High hydrostatic pressure (HHP) has been employed to gelatinize or physically modify starch dispersions. In this study, waxy maize starch, normal maize starch, and two high amylose content starch were processed by a HHP of the order of 600 MPa, at 25°C for 15min. The effect of HHP processing on the crystallization of maize starches with various amylose content during storage at 4°C was investigated. Crystallization kinetics of HHP treated starch gels were investigated using rheology and FTIR. The effect of crystallization on the mechanical properties of starch gel network were evaluated in terms of dynamic complex modulus (G*). The crystallization induced increase of short-range helices structures were investigated using FTIR. The pressure releasing rate does not affect the starch retrogradation behaviour. The rate and extent of retrogradation depends on the amylose content of amylose starch. The least retrogradation was observed in HHP treated waxy maize starch. The rate of retrogradation is higher for HHP treated high amylose maize starch than that of normal maize starch. A linear relationship between the extent of retrogradation (phase distribution) measured by FTIR and G* is proposed.

  9. Retrogradation of Maize Starch after High Hydrostatic Pressure Gelation: Effect of Amylose Content and Depressurization Rate.

    Science.gov (United States)

    Yang, Zhi; Swedlund, Peter; Gu, Qinfen; Hemar, Yacine; Chaieb, Sahraoui

    2016-01-01

    High hydrostatic pressure (HHP) has been employed to gelatinize or physically modify starch dispersions. In this study, waxy maize starch, normal maize starch, and two high amylose content starch were processed by a HHP of the order of 600 MPa, at 25°C for 15min. The effect of HHP processing on the crystallization of maize starches with various amylose content during storage at 4°C was investigated. Crystallization kinetics of HHP treated starch gels were investigated using rheology and FTIR. The effect of crystallization on the mechanical properties of starch gel network were evaluated in terms of dynamic complex modulus (G*). The crystallization induced increase of short-range helices structures were investigated using FTIR. The pressure releasing rate does not affect the starch retrogradation behaviour. The rate and extent of retrogradation depends on the amylose content of amylose starch. The least retrogradation was observed in HHP treated waxy maize starch. The rate of retrogradation is higher for HHP treated high amylose maize starch than that of normal maize starch. A linear relationship between the extent of retrogradation (phase distribution) measured by FTIR and G* is proposed.

  10. Retrogradation of Maize Starch after High Hydrostatic Pressure Gelation: Effect of Amylose Content and Depressurization Rate.

    Directory of Open Access Journals (Sweden)

    Zhi Yang

    Full Text Available High hydrostatic pressure (HHP has been employed to gelatinize or physically modify starch dispersions. In this study, waxy maize starch, normal maize starch, and two high amylose content starch were processed by a HHP of the order of 600 MPa, at 25°C for 15min. The effect of HHP processing on the crystallization of maize starches with various amylose content during storage at 4°C was investigated. Crystallization kinetics of HHP treated starch gels were investigated using rheology and FTIR. The effect of crystallization on the mechanical properties of starch gel network were evaluated in terms of dynamic complex modulus (G*. The crystallization induced increase of short-range helices structures were investigated using FTIR. The pressure releasing rate does not affect the starch retrogradation behaviour. The rate and extent of retrogradation depends on the amylose content of amylose starch. The least retrogradation was observed in HHP treated waxy maize starch. The rate of retrogradation is higher for HHP treated high amylose maize starch than that of normal maize starch. A linear relationship between the extent of retrogradation (phase distribution measured by FTIR and G* is proposed.

  11. A novel power-efficient high-speed clock management unit using quantum-dot cellular automata

    Science.gov (United States)

    Abutaleb, M. M.

    2017-04-01

    Quantum-dot cellular automata (QCA) is one of the most attractive alternatives for complementary metal-oxide semiconductor technology. The QCA widely supports a new paradigm in the field of nanotechnology that has the potential for high density, low power, and high speed. The clock manager is an essential building block in the new microwave and radio frequency integrated circuits. This paper describes a novel QCA-based clock management unit (CMU) that provides innovative clocking capabilities. The proposed CMU is achieved by utilizing edge-triggered D-type flip-flops (D-FFs) in the design of frequency synthesizer and phase splitter. Edge-triggered D-FF structures proposed in this paper have the successful QCA implementation and simulation with the least complexity and power dissipation as compared to earlier structures. The frequency synthesizer is used to generate new clock frequencies from the reference clock frequency based on a combination of power-of-two frequency dividers. The phase splitter is integrated with the frequency synthesizer to generate four clock signals that are 90o out of phase with each other. This paper demonstrates that the proposed QCA CMU structure has a superior performance. Furthermore, the proposed CMU is straightforwardly scalable due to the use of modular component architecture.

  12. An improved Cellular Automata model to simulate the behavior of high density crowd and validation by experimental data

    Science.gov (United States)

    Feliciani, Claudio; Nishinari, Katsuhiro

    2016-06-01

    In this article we present an improved version of the Cellular Automata floor field model making use of a sub-mesh system to increase the maximum density allowed during simulation and reproduce phenomena observed in dense crowds. In order to calibrate the model's parameters and to validate it we used data obtained from an empirical observation of bidirectional pedestrian flow. A good agreement was found between numerical simulation and experimental data and, in particular, the double outflow peak observed during the formation of deadlocks could be reproduced in numerical simulations, thus allowing the analysis of deadlock formation and dissolution. Finally, we used the developed high density model to compute the flow-ratio dependent fundamental diagram of bidirectional flow, demonstrating the instability of balanced flow and predicting the bidirectional flow behavior at very high densities. The model we presented here can be used to prevent dense crowd accidents in the future and to investigate the dynamics of the accidents which already occurred in the past. Additionally, fields such as granular and active matter physics may benefit from the developed framework to study different collective phenomena.

  13. Versatile assays for high throughput screening for activators or inhibitors of intracellular proteases and their cellular regulators.

    Directory of Open Access Journals (Sweden)

    Hideki Hayashi

    Full Text Available Intracellular proteases constitute a class of promising drug discovery targets. Methods for high throughput screening against these targets are generally limited to in vitro biochemical assays that can suffer many technical limitations, as well as failing to capture the biological context of proteases within the cellular pathways that lead to their activation. METHODS #ENTITYSTARTX00026;We describe here a versatile system for reconstituting protease activation networks in yeast and assaying the activity of these pathways using a cleavable transcription factor substrate in conjunction with reporter gene read-outs. The utility of these versatile assay components and their application for screening strategies was validated for all ten human Caspases, a family of intracellular proteases involved in cell death and inflammation, including implementation of assays for high throughput screening (HTS of chemical libraries and functional screening of cDNA libraries. The versatility of the technology was also demonstrated for human autophagins, cysteine proteases involved in autophagy.Altogether, the yeast-based systems described here for monitoring activity of ectopically expressed mammalian proteases provide a fascile platform for functional genomics and chemical library screening.

  14. New formulations of sunflower based bio-lubricants with high oleic acid content – VOSOLUB project

    Directory of Open Access Journals (Sweden)

    Leao J. D.

    2016-09-01

    Full Text Available VOSOLUB project is a demonstration project supported by Executive Agency for Small and Medium-sized Enterprises (EASME that aims at testing under real operating conditions new formulations of sunflower-based biolubricants with high oleic acid content. These biolubricant formulations (including hydraulic fluids, greases, and neat oil metal-working fluids will be tested in three European demonstrating sites. Their technical performance will be evaluated and compared to corresponding mineral lubricants ones. In order to cover the demand for the sunflower base oil, a European SMEs network will be established to ensure the supply of the base at a competitive market price. Results presented concerns the base oil quality confirmed to be in accordance with the specification required, in particular on Free Fatty acid content, Phosphorus content, rancimat induction time and oleic acid content (ITERG. The oil characteristics specific for lubricant application analyzed by BfB Oil Research under normalized methods, match with lubricant specifications requirement such as viscosity, cold & hot properties, surface properties, anti-oxidant properties and thermal stability, anti-wear and EP properties, anti-corrosion properties Performance of the new biolubricant have been assessed by formulators and TEKNIKER First results on the use of new lubricant on real condition for rail Grease (produced by RS CLARE and tested with Sheffield Supertram, Hydraulic oil (produced by BRUGAROLAS and cutting oil (produced by MOTUL TECH and tested with innovative machining, turning are described.

  15. Analysis of the origins of content non-uniformity in high-shear wet granulation.

    Science.gov (United States)

    Oka, Sarang; Smrčka, David; Kataria, Anjali; Emady, Heather; Muzzio, Fernando; Štěpánek, František; Ramachandran, Rohit

    2017-08-07

    In this study, the origins of granule content non-uniformity in the high-shear wet granulation of a model two-component pharmaceutical blend were investigated. Using acetaminophen as the active pharmaceutical ingredient (API) and microcrystalline cellulose as the excipient, the distribution of the API across the granule size classes was measured for a range of conditions that differed in the duration of the initial dry mixing stage, the overall composition of the blend and the wet massing time. The coarse granule fractions were found to be systematically sub-potent, while the fines were enriched in the API. The extent of content non-uniformity was found to be dependent on two factors - powder segregation during dry mixing and redistribution of the API between the granule size fractions during the wet massing phase. The latter was demonstrated in an experiment where the excipient was pre-granulated, the API was added later and wet massed. The content non-uniformity in this case was comparable to that obtained when both components were present in the granulator from the beginning. With increasing wet massing time, the extent of content non-uniformity decreased, indicating that longer wet massing times might be a solution for systems with a natural tendency for component segregation. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Predicting In Vivo Anti-Hepatofibrotic Drug Efficacy Based on In Vitro High-Content Analysis

    OpenAIRE

    2011-01-01

    BACKGROUND/AIMS: Many anti-fibrotic drugs with high in vitro efficacies fail to produce significant effects in vivo. The aim of this work is to use a statistical approach to design a numerical predictor that correlates better with in vivo outcomes. METHODS: High-content analysis (HCA) was performed with 49 drugs on hepatic stellate cells (HSCs) LX-2 stained with 10 fibrotic markers. ~0.3 billion feature values from all cells in >150,000 images were quantified to reflect the drug effects. A sy...

  17. High-throughput analysis of total nitrogen content that replaces the classic Kjeldahl method.

    Science.gov (United States)

    Yasuhara, T; Nokihara, K

    2001-10-01

    A high-throughput method for determination of total nitrogen content has been developed. The method involves decomposition of samples, followed by trapping and quantitative colorimetric determination of the resulting ammonia. The present method is rapid, facile, and economical. Thus, it can replace the classic Kjeldahl method through its higher efficiency for determining multiple samples. Compared to the classic method, the present method is economical and environmentally friendly. Based on the present method, a novel reactor was constructed to realize routine high-throughput analyses of multiple samples such as those found for pharmaceutical materials, foods, and/or excrements.

  18. Electrical resistance stability of high content carbon fiber reinforced cement composite

    Institute of Scientific and Technical Information of China (English)

    YANG Zai-fu; TANG Zu-quan; LI Zhuo-qiu; QIAN Jue-shi

    2005-01-01

    The influences of curing time, the content of free evaporable water in cement paste, environmental temperature, and alternative heating and cooling on the electrical resistance of high content carbon fiber reinforced cement (CFRC) paste are studied by experiments with specimens of Portland cement 42.5 with 10 mm PAN-based carbon fiber and methylcellulose. Experimental results indicate that the electrical resistance of CFRC increases relatively by 24% within a hydration time of 90 d and almost keeps constant after 14 d, changes hardly with the mass loss of free evaporable water in the concrete dried at 50℃C, increases relatively by 4% when ambient temperature decreases from 15℃ to-20℃, and decreases relatively by 13% with temperature increasing by 88℃. It is suggested that the electric resistance of the CFRC is stable, which is testified by the stable power output obtained by electrifying the CFRC slab with a given voltage. This implies that such kind of high content carbon fiber reinforced cement composite is potentially a desirable electrothermal material for airfield runways and road surfaces deicing.

  19. Effect of skin care with an emollient containing a high water content on mild uremic pruritus.

    Science.gov (United States)

    Okada, Kazuyoshi; Matsumoto, Koichi

    2004-10-01

    Skin care is very important for preventing uremic pruritus. However, mild uremic pruritus has usually been treated with antihistamine and urea-containing ointments. We therefore examined the effects of an aqueous gel with higher water content. Twenty hemodialysis patients with mild pruritus who were not being treated with any emollient were divided into two groups of 10 each. Patients in one group were treated with an aqueous gel containing 80% water. This emollient was applied twice daily for 2 weeks. No emollient was applied for the next 2 weeks. The other group of patients were not treated with any emollient for the 4 weeks. Visual analog scale scores for itching in the experimental group at week 2 were significantly decreased compared with that at week 0 (3.5 +/- 0.3 vs 0.6 +/- 0.2, P emollient was stopped. There were no significant changes in the control group during the study. It is concluded that the aqueous gel with high water content reduced itching and improved xerosis in patients with mild uremic pruritus. It is reasonable that skin care with an emollient containing a high water content is first started for hemodialysis patients with xerosis, even if they do not feel itching.

  20. Pulsed Nd:YAG laser cladding of high silicon content coating on low silicon steel

    Institute of Scientific and Technical Information of China (English)

    Danyang Dong; Changsheng Liu; Bin Zhang; Jun Miao

    2007-01-01

    A pulsed Nd:YAG (yttrium aluminum garnet) laser-based technique was employed to clad low silicon steel with preplaced Si and Fe mixed powders for high Si content. The surface morphology, microstructural evolution, phase composition, and Si distribution,within the obtained cladding coatings, were characterized by optical microscopy (OM), field emission scanning electron microscopy (FE-SEM), with associated energy dispersive spectroscopy (EDS), transmission electron microscopy (TEM), and X-ray diffraction (XRD). The microhardness was also measured along the depth direction of the specimens. A crack- and pore-free cladding coating through excellent metallurgical bonding with the substrate was successfully prepared on low silicon steel by means of optimized single-track and multi-track laser cladding. The phases of the coating are α-Fe, γ-Fe, and FeSi. The high microhardness of the lasercladding zone is considered as an increase in Si content and as the refined microstructure produced by the laser treatment. The Si contents of the cladding coatings were about 5.8wt% in the single-track cladding and 6.5wt% in the multi-track cladding, respectively.

  1. Utilization of potato starch processing wastes to produce animal feed with high lysine content.

    Science.gov (United States)

    Li, Ying; Liu, Bingnan; Song, Jinzhu; Jiang, Cheng; Yang, Qian

    2015-02-01

    This work aims to utilize wastes from the potato starch industry to produce single-cell protein (SCP) with high lysine content as animal feed. In this work, S-(2-aminoethyl)-L-cysteine hydrochloride-resistant Bacillus pumilus E1 was used to produce SCP with high lysine content, whereas Aspergillus niger was used to degrade cellulose biomass and Candida utilis was used to improve the smell and palatability of the feed. An orthogonal design was used to optimize the process of fermentation for maximal lysine content. The optimum fermentation conditions were as follows: temperature of 40°C, substrate concentration of 3%, and natural pH of about 7.0. For unsterilized potato starch wastes, the microbial communities in the fermentation process were determined by terminal restriction fragment length polymorphism analysis of bacterial 16S rRNA genes. Results showed that the dominant population was Bacillus sp. The protein quality as well as the amino acid profile of the final product was found to be significantly higher compared with the untreated waste product at day 0. Additionally, acute toxicity test showed that the SCP product was non-toxic, indicating that it can be used for commercial processing.

  2. High Water Contents in the Siberian Cratonic Mantle: An FTIR Study of Udachnaya Peridotite Xenoliths

    Science.gov (United States)

    Doucet, Luc S.; Peslier, Anne H.; Ionov, Dimitri A.; Brandon, Alan D.; Golovin, Alexander V.; Ashchepkov, Igor V.

    2013-01-01

    Water is believed to be a key factor controlling the long-term stability of cratonic lithosphere, but mechanisms responsible for the water content distribution in the mantle remain poorly constrained. Water contents were obtained by FTIR in olivine, pyroxene and garnet for 20 well-characterized peridotite xenoliths from the Udachnaya kimberlite (central Siberian craton) and equilibrated at 2-7 GPa. Water contents in minerals do not appear to be related to interaction with the host kimberlite. Diffusion modeling indicates that the core of olivines preserved their original water contents. The Udachnaya peridotites show a broad range of water contents in olivine (6.5 +/- 1.1 to 323 +- 65 ppm H2O (2 sigma)), and garnet (0 - 23 +/- 6 ppm H2O). The water contents of olivine and garnet are positively correlated with modal clinopyroxene, garnet and FeO in olivine. Water-rich garnets are also rich in middle rare earth elements. This is interpreted as the result of interaction between residual peridotites and water rich-melts, consistent with modal and cryptic metasomatism evidenced in the Siberian cratonic mantle. The most water-rich Udachnaya minerals contain 2 to 3 times more water than those from the Kaapvaal craton, the only craton with an intact mantle root for which water data is available. The highest water contents in olivine and orthopyroxene in this study (>= 300 ppm) are found at the bottom of the lithosphere (> 6.5 GPa). This is in contrast with the Kaapvaal craton where the olivines of peridotites equilibrated at > 6.4 GPa have 6 GPa is lower or similar (8.4× 10(exp 16) to 8.0× 10(exp 18) Pa./s) to that of the asthenosphere (<= 3.7x10(exp 18) Pa./s ). Such lithologies would not be able to resist delamination by the convecting asthenosphere. However, seismology studies as well as the high equilibration pressures of our samples indicate that the Udachnaya cratonic lithosphere is 220-250 km thick. Consequently, the water-rich peridotites are likely not

  3. Recent advances in quantitative high throughput and high content data analysis.

    Science.gov (United States)

    Moutsatsos, Ioannis K; Parker, Christian N

    2016-01-01

    High throughput screening has become a basic technique with which to explore biological systems. Advances in technology, including increased screening capacity, as well as methods that generate multiparametric readouts, are driving the need for improvements in the analysis of data sets derived from such screens. This article covers the recent advances in the analysis of high throughput screening data sets from arrayed samples, as well as the recent advances in the analysis of cell-by-cell data sets derived from image or flow cytometry application. Screening multiple genomic reagents targeting any given gene creates additional challenges and so methods that prioritize individual gene targets have been developed. The article reviews many of the open source data analysis methods that are now available and which are helping to define a consensus on the best practices to use when analyzing screening data. As data sets become larger, and more complex, the need for easily accessible data analysis tools will continue to grow. The presentation of such complex data sets, to facilitate quality control monitoring and interpretation of the results will require the development of novel visualizations. In addition, advanced statistical and machine learning algorithms that can help identify patterns, correlations and the best features in massive data sets will be required. The ease of use for these tools will be important, as they will need to be used iteratively by laboratory scientists to improve the outcomes of complex analyses.

  4. Iodine content of infant formulas and iodine intake of premature babies: high risk of iodine deficiency.

    Science.gov (United States)

    Ares, S; Quero, J; Durán, S; Presas, M J; Herruzo, R; Morreale de Escobar, G

    1994-11-01

    As part of a study of thyroid function in premature babies, the iodine content of their mothers' breast milk, that of 32 formulas from different brands used in Spain, and that of 127 formulas used in other countries was determined. Breast milk contained more iodine--mean (SEM) 10 (1) microgram/dl--than most of the formulas, especially those for premature babies. Iodine intakes were therefore below the recommended daily amount (RDA) for newborns: babies of 27-30 weeks' gestational age took 3.1 (1.1) micrograms/day at 5 days of age and 29.8 (2.7) micrograms by 2 months of age. This problem is not exclusive to Spanish premature babies as the iodine content of many of the formulas on sale in other countries was also inadequate. It is concluded that preterm infants who are formula fed are at high risk of iodine deficiency.

  5. Data and prediction of water content of high pressure nitrogen, methane and natural gas

    DEFF Research Database (Denmark)

    Folas, Georgios; Froyna, E.W.; Lovland, J.;

    2007-01-01

    New data for the equilibrium water content of nitrogen, methane and one natural gas mixture are presented. The new binary data and existing binary sets were compared to calculated values of dew point temperature using both the CPA (Cubic-Plus-Association) EoS and the GERG-water EoS. CPA is purely...... predictive (i.e. all binary interaction parameters are set equal to 0), while GERG-water uses a temperature dependent interaction parameter fitted to published data. The GERG-water model is proposed as an ISO standard for determining the water content of natural gas. The data sets for nitrogen cover...... they have large scatter. The data sets that have been measured at low pressures extrapolate well towards the ideal equilibrium values. The two models show similar results, but differ at high pressure and/or temperature. CPA is shown to extrapolate well for methane-water to 1000 bar and 573 K, and our...

  6. Development of High Strength Steel Designed for Carbonitriding with High Nitrogen Content to Be Used for Automatic Transmission Gears

    Institute of Scientific and Technical Information of China (English)

    Youichi Watanabe

    2004-01-01

    To downsize and lighten automatic transmission components, the gears installed must be strengthened in terms of pitting endurance and/or wear resistance. The most important metallurgical factor affecting fractures is well known to be resistance to softening when steel is tempered at approximately 573 K. Carbonitriding with a high anount of nitrogen is a very effective production technique because nitrogen increases the resistance during tempering. However, structural anomalies begin to appear in the surface layer when the nitrogen content exceeds 0.6 mass% in the chromium steel generally used. To address this, we have developed new high-strength chromium steel with an optimized chemical composition that effectively inhibits anomalies even when carbonitriding with a nitrogen content of more than 0.6 mass%.We performed a drivetrain durability test on an automatic transmission component designed to have excellent contact fatigue strength and a tooth root bending impact and fatigue strength. We found that the developed steel that was carbonitrided with a content of about 0.9 mass%, and then shot peened hard, has a pitting life of roughly 4.5 times that of conventionally manufactured steel.

  7. High N-content a-C:N films elaborated by femtosecond PLD with plasma assistance

    Energy Technology Data Exchange (ETDEWEB)

    Maddi, C. [Université de Lyon, F-69003, Lyon, France, Université de Saint-Étienne, Laboratoire Hubert Curien (UMR 5516 CNRS) , 42000 Saint-Étienne (France); Donnet, C., E-mail: Christophe.Donnet@univ-st-etienne.fr [Université de Lyon, F-69003, Lyon, France, Université de Saint-Étienne, Laboratoire Hubert Curien (UMR 5516 CNRS) , 42000 Saint-Étienne (France); Loir, A.-S.; Tite, T. [Université de Lyon, F-69003, Lyon, France, Université de Saint-Étienne, Laboratoire Hubert Curien (UMR 5516 CNRS) , 42000 Saint-Étienne (France); Barnier, V. [Laboratoire Georges Friedel, Ecole Nationale Supérieure des Mines, 42023 Saint-Etienne (France); Rojas, T.C.; Sanchez-Lopez, J.C. [Instituto de Ciencia de Materiales de Sevilla (CSIC-US) , Avda. Américo Vespucio 49, 41092 Sevilla (Spain); Wolski, K. [Laboratoire Georges Friedel, Ecole Nationale Supérieure des Mines, 42023 Saint-Etienne (France); Garrelie, F. [Université de Lyon, F-69003, Lyon, France, Université de Saint-Étienne, Laboratoire Hubert Curien (UMR 5516 CNRS) , 42000 Saint-Étienne (France)

    2015-03-30

    Graphical abstract: - Highlights: • Nitrogen doped amorphous carbon films were deposited by DC reactive plasma femtosecond (fs) -PLD and conventional fs-PLD. • High nitrogen content in plasma assisted films. • More ordered sp2 rich graphitic clusters both in terms of structural and topological order. • Correlation length La of the clusters increases with nitrogen incorporation. • Formation of CN bonds at the expense of CC bonds with N content. • At the highest nitrogen concentration, terminal C≡N groups are incorporated in the film. • Correlation between film composition and plasma process. - Abstract: Amorphous carbon nitride (a-C:N) thin films are a interesting class of carbon-based electrode materials. Therefore, synthesis and characterization of these materials have found lot of interest in environmental analytical microsystems. Herein, we report the nitrogen-doped amorphous carbon thin film elaboration by femtosecond pulsed laser deposition (fs-PLD) both with and without a plasma assistance. The chemical composition and atomic bonding configuration of the films were investigated by multi-wavelength (MW) Raman spectroscopy, X-ray photoelectron spectroscopy (XPS) and electron energy-loss spectroscopy (EELS). The highest nitrogen content, 28 at.%, was obtained with plasma assistance. The I(D)/I(G) ratio and the G peak position increased as a function of nitrogen concentration, whereas the dispersion and full width at half maximum (FWHM) of G peak decreased. This indicates more ordered graphitic like structures in the films both in terms of topological and structural, depending on the nitrogen content. EELS investigations were correlated with MW Raman results. The interpretation of XPS spectra of carbon nitride films remains a challenge. Plasma assisted PLD in the femtosecond regime led to a significant high nitrogen concentration, which is highlighted on the basis of collisional processes in the carbon plasma plume interacting with the nitrogen

  8. Microspheres with an ultra high holmium content for brachytherapy of malignancies

    Energy Technology Data Exchange (ETDEWEB)

    Lira, Raphael A.; Myamoto, Douglas M.; Souza, Jaime R.; Nascimento, Nanci; Azevedo, Mariangela de Burgos M. de [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil). Centro de Biotecnologia; Osso Junior, Joao A. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil). Centro de Radiofarmacia; Martinelli, Jose R. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil). Centro de Ciencias e Tecnologia de Materiais

    2011-07-01

    The overall objective of this work is to develop biodegradable microspheres intended for internal radiation therapy which provides an improved treatment for hepatic carcinomas. The most studied brachytherapy system employing microspheres made of holmium-biopolymer system is composed by poly(L-lactic acid) (PLLA) and holmium acetylacetonate (HoAcAc). The importance of the holmium high content in the microspheres can be interpreted as follow from a therapeutic standpoint, to achieve an effective use of microspheres loaded with HoAcAc, a high content of holmium is required to yield enough radioactivity with a relatively low amount of microspheres.The usual amounts of holmium that are incorporated in the microspheres composed by poly(L-lactic acid) and HoAcAc are 17.0 {+-} 0.5% (w/w) of holmium, which corresponds to a loading of about 50% of HoAcAc. Different approaches have been investigated to increase that value. One updated approach towards this direction is the production of microspheres with ultrahigh holmium as matrix using HoAcAc crystals as the sole starting material without the use of biopolymer. Likewise, in the search of microspheres with increased holmium content , it has been demonstrated that by changing the HoAcAc crystal structure by its recrystallization from crystal phase to the amorphous there is lost of acetylacetonate and water molecules causing the increasing of the holmium content. Microspheres were prepared by solvent evaporation, using holmium acetylacetonate (HoAcAc) crystals as the sole ingredient. Microspheres were characterized by using light and scanning electron microscopy, infrared and Raman spectroscopy, differential scanning calorimetry, X-rays diffraction, and confocal laser scanning microscopy. (author)

  9. A Place for Content Literacy: Incorporating Vocabulary and Comprehension Strategies in the High School Science Classroom

    Science.gov (United States)

    Misulis, Katherine E.

    2011-01-01

    To help students learn and apply science content, teachers can embed content literacy instruction within science instruction. This involves teaching the content and the literacy skills students need to learn that content, such as vocabulary and comprehension. In this article, the author provides tips on how to incorporate content literacy…

  10. Foraminiferal indicators of high OC content in marine sediments; a question of OC source?

    Science.gov (United States)

    Lo Giudice Cappelli, Elena; Austin, William; Smith, Laura; Ward, Hannah; Clarke, Jess; Green, Jade; Smeaton, Craig

    2017-04-01

    West Shetland voes (sea inlets), on Scotland northern maritime periphery, represent a range of environmental and depositional settings in an area of coastal ocean which is characterised by a generally high organic carbon content in marine sediments. Possible sources of organic carbon are fish farms, as this region is experiencing a large expansion in marine aquaculture, and/or erosion of peatland. Twenty-one sites in six voes were sampled in August 2015 following an organic carbon gradient. The aims of this study are: 1) to establish the modern biogeography of benthic foraminifera in west Shetland voes, 2) to explore the use of these organisms as bio-indicators of high organic carbon content in marine sediments, and 3) to trace the source of organic carbon and its transportation between different habitats and pools (e.g.: terrestrial to marine; marine aquaculture to sediments). To achieve this, we paired together spatial variations in the composition of benthic foraminiferal assemblages (species abundance and diversity) with changes in the physical properties of marine sediments determined by carbon stable isotope measurements and loss on ignition analysis. Preliminary results show a positive relationship between high abundance of the agglutinated benthic foraminifera Eggerelloides scaber, high organic carbon content in the sediments and proximity to fish farms. Although aquaculture farming in these areas may be a contributing factor, radiocarbon dates demonstrate that organic enrichment in the sediments pre-dates the introduction of aquaculture, while carbon stable isotope measurements point to peat erosion as a more likely source of organic carbon in west Shetland voes. Therefore, peatland erosion is a significant source of organic carbon in marine sediments and may be an important contributor to the store of blue carbon in west Shetland voes and, more in general, in the coastal ocean.

  11. Identification of host cellular proteins that interact with the M protein of a highly pathogenic porcine reproductive and respiratory syndrome virus vaccine strain.

    Science.gov (United States)

    Wang, Qian; Li, Yanwei; Dong, Hong; Wang, Li; Peng, Jinmei; An, Tongqing; Yang, Xufu; Tian, Zhijun; Cai, Xuehui

    2017-02-22

    The highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) continues to pose one of the greatest threats to the swine industry. M protein is the most conserved and important structural protein of PRRSV. However, information about the host cellular proteins that interact with M protein remains limited. Host cellular proteins that interact with the M protein of HP-PRRSV were immunoprecipitated from MARC-145 cells infected with PRRSV HuN4-F112 using the M monoclonal antibody (mAb). The differentially expressed proteins were identified by LC-MS/MS. The screened proteins were used for bioinformatics analysis including Gene Ontology, the interaction network, and the enriched KEGG pathways. Some interested cellular proteins were validated to interact with M protein by CO-IP. The PRRSV HuN4-F112 infection group had 10 bands compared with the control group. The bands included 219 non-redundant cellular proteins that interact with M protein, which were identified by LC-MS/MS with high confidence. The gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) pathway bioinformatic analyses indicated that the identified proteins could be assigned to several different subcellular locations and functional classes. Functional analysis of the interactome profile highlighted cellular pathways associated with protein translation, infectious disease, and signal transduction. Two interested cellular proteins-nuclear factor of activated T cells 45 kDa (NF45) and proliferating cell nuclear antigen (PCNA)-that could interact with M protein were validated by Co-IP and confocal analyses. The interactome data between PRRSV M protein and cellular proteins were identified and contribute to the understanding of the roles of M protein in the replication and pathogenesis of PRRSV. The interactome of M protein will aid studies of virus/host interactions and provide means to decrease the threat of PRRSV to the swine industry in the future.

  12. New Literacies and Pedagogical Migration: Exploring Impacts of New Literacies on Instruction and Learning in High School Content Classrooms

    Science.gov (United States)

    Mooney, Melody Anne

    2013-01-01

    The purpose of this study is to examine the impacts of integrating new literacies practices into high school curriculum content classrooms. This study examined data collected through observations and semi-structured interviews conducted with four high school content curriculum teachers. Analysis of the data resulted in essential findings emerging…

  13. Molecular Assisted Breeding and Adaptability Analysis of Zhongyouza 11 with Super High Oil Content

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Using a three-line system of Shaan 2A cytoplasmic male sterility, a hybrid variety rapeseed, Zhongyouza 11, was developed by the combination of traditional methods and a molecular marker-assisted breeding technique. This hybrid variety had high grain yield, high oil content, wide adaptability, and good quality. The oil content of Zhongyouza 11 is steadily above 43% in the three ecological regions along the Yangtze River valley, which is the largest winter oilseed production area in China. The highest oil content of Zhongyouza 11 reached 46.68% in the upper reaches of the Yangtze River valley in 2003-2004, which was the highest among all lines involved in the national regional rapeseed variety trials. In the regional yield trials in Hubei Province, the average yield of Zhongyouza 11 reached 2 853 kg ha-1 (2002-2004), yielding more than the control variety Zhongshuang 6 by 11.34%. In the national trials, the yield of Zhongyouza 11 reached 2 405.7, 2 697.3, and 2 770.2 kg ha-1 in the upper, middle, and lower reaches of the Yangtze River valley, yielding more than the control varieties by 11.52, 12.9, and 14.92%, respectively, in 2003-2005. Both the high oil content and high grain yield contributed to the high oil yield of Zhongyouza 11. The oil yield of Zhongyouza 11 in the national trials reached 1083.3, 1210.2, and 1224.8 kg ha-1, yielding more than the controls by 29.42, 27.65, and 20.98%, respectively, in the upper, middle, and lower reaches of the Yangtze River valley. The highest oil yield reached 1369.65 kg ha-1 in the middle reach of the Yangtze River valley(2003-2004), with an increase of 46.21% compared to the control. The experimental sites distributed widely along the Yangtze River valley, cover most of the Chinese winter rapeseed production areas. Among the 89 experimental location/years, the yield of Zhongyouza 11 surpassed the controls at 77 location/years in the trials, indicating its wide adaptability.Zhongyouza 11 was the only one that passed the

  14. High phosphorylase activity is correlated with increased potato minituber formation and starch content during extended clinorotation

    Science.gov (United States)

    Nedukha, O. M.; Schnyukova, E. I.; Leach, J. E.

    2003-05-01

    The major purpose of these experiments were to investigate growth of potato storage organs and starch synthesis in minitubers at slow horizontal clinorotation (2 rpm), which partly mimics microgravity, and a secondary goal was to study the activity and localization of phosphorylase (EC 2.4.1.1) in storage parenchyma under these conditions. Miniplants of Solanum tuberosum L. (cv Adreta) were grown in culture for 30 days for both the vertical control and the horizontal clinorotation. During long-term clinorotation, an acceleration of minituber formation, and an increase of amyloplast number and size in storage parenchyma cells, as well as increased starch content, was observed in the minitubers. The differences among cytochemical reaction intensity, activity of phosphorylase, and carbohydrate content in storage parenchyma cells of minitubers grown in a horizontal clinostat were established by electron-cytochemical and biochemical methods. It is shown that high phosphorylase activity is correlated with increased starch content during extended clinorotation. The results demonstrate the increase in carbohydrate metabolism and possible accelerated growth of storage organs under the influence of microgravity, as mimicked by clinorotation; therefore, clinorotation can be used as a basis for future studies on mechanisms of starch synthesis under microgravity.

  15. Thermogravimetric Analysis of Effects of High-Content Limstone Addition on Combustion Characteristics of Taixi Anthracite

    Institute of Scientific and Technical Information of China (English)

    ZHANG Hong; LI Mei; SUN Min; WEI Xian-yong

    2004-01-01

    Combustion characteristics of Taixi anthracite admixed with high content of limestone addition were investigated with thermogravimetric analysis. The results show that limestone addition has a little promoting effect on the ignition of raw coals as a whole. The addition of limestone is found to significantly accelerate the combustion and burnout of raw coals. The higher the sample mass is, the more significant the effect will be. The results also show that the change of limestone proportion between 45%-80% has little effect on ignition temperatures of coal in the blended samples. Increasing limestone content lowers the temperature corresponding to the maximum weight loss. Although higher maximum mass loss rates are observed with higher limestone content, the effect is found not ascribed to changing limestone addition, but to the decrease of absolute coal mass in the sample. The change of limestone proportion has little effect on its burnout temperature. Mechanism analysis indicates that these phenomena result mainly from improved heat conduction due to limestone addition.

  16. Biodiesel from the seed oil of Treculia africana with high free fatty acid content

    Energy Technology Data Exchange (ETDEWEB)

    Adewuyi, Adewale [Redeemer' s University, Department of Chemical Sciences, Faculty of Natural Sciences, Redemption City, Ogun State (Nigeria); Oderinde, Rotimi A.; Ojo, David F.K. [University of Ibadan, Industrial Unit, Department of Chemistry, Ibadan, Oyo State (Nigeria)

    2012-12-15

    Oil was extracted from the seed of Treculia africana using hexane. The oil was characterized and used in the production of biodiesel. Biodiesel was produced from the seed oil of T. africana using a two-step reaction system. The first step was a pretreatment which involved the use of 2 % sulfuric acid in methanol, and secondly, transesterification reaction using KOH as catalyst. Saponification value of the oil was 201.70 {+-} 0.20 mg KOH/g, free fatty acid was 8.20 {+-} 0.50 %, while iodine value was 118.20 {+-} 0.50 g iodine/100 g. The most dominant fatty acid was C18:2 (44 %). The result of the method applied showed a conversion which has ester content above 98 %, flash point of 131 {+-} 1.30 C, and phosphorus content below 1 ppm in the biodiesel. The biodiesel produced exhibited properties that were in agreement with the European standard (EN 14214). This study showed that the high free fatty acid content of T. africana seed oil can be reduced in a one-step pretreatment of esterification reaction using H{sub 2}SO{sub 4} as catalyst. (orig.)

  17. Factor analysis in optimization of formulation of high content uniformity tablets containing low dose active substance.

    Science.gov (United States)

    Lukášová, Ivana; Muselík, Jan; Franc, Aleš; Goněc, Roman; Mika, Filip; Vetchý, David

    2017-09-11

    Warfarin is intensively discussed drug with narrow therapeutic range. There have been cases of bleeding attributed to varying content or altered quality of the active substance. Factor analysis is useful for finding suitable technological parameters leading to high content uniformity of tablets containing low amount of active substance. The composition of tabletting blend and technological procedure were set with respect to factor analysis of previously published results. The correctness of set parameters was checked by manufacturing and evaluation of tablets containing 1-10mg of warfarin sodium. The robustness of suggested technology was checked by using "worst case scenario" and statistical evaluation of European Pharmacopoeia (EP) content uniformity limits with respect to Bergum division and process capability index (Cpk). To evaluate the quality of active substance and tablets, dissolution method was developed (water; EP apparatus II; 25rpm), allowing for statistical comparison of dissolution profiles. Obtained results prove the suitability of factor analysis to optimize the composition with respect to batches manufactured previously and thus the use of metaanalysis under industrial conditions is feasible. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Mutational breeding and genetic engineering in the development of high grain protein content.

    Science.gov (United States)

    Wenefrida, Ida; Utomo, Herry S; Linscombe, Steve D

    2013-12-04

    Cereals are the most important crops in the world for both human consumption and animal feed. Improving their nutritional values, such as high protein content, will have significant implications, from establishing healthy lifestyles to helping remediate malnutrition problems worldwide. Besides providing a source of carbohydrate, grain is also a natural source of dietary fiber, vitamins, minerals, specific oils, and other disease-fighting phytocompounds. Even though cereal grains contain relatively little protein compared to legume seeds, they provide protein for the nutrition of humans and livestock that is about 3 times that of legumes. Most cereal seeds lack a few essential amino acids; therefore, they have imbalanced amino acid profiles. Lysine (Lys), threonine (Thr), methionine (Met), and tryptophan (Trp) are among the most critical and are a limiting factor in many grain crops for human nutrition. Tremendous research has been put into the efforts to improve these essential amino acids. Development of high protein content can be outlined in four different approaches through manipulating seed protein bodies, modulating certain biosynthetic pathways to overproduce essential and limiting amino acids, increasing nitrogen relocation to the grain through the introduction of transgenes, and exploiting new genetic variance. Various technologies have been employed to improve protein content including conventional and mutational breeding, genetic engineering, marker-assisted selection, and genomic analysis. Each approach involves a combination of these technologies. Advancements in nutrigenomics and nutrigenetics continue to improve public knowledge at a rapid pace on the importance of specific aspects of food nutrition for optimum fitness and health. An understanding of the molecular basis for human health and genetic predisposition to certain diseases through human genomes enables individuals to personalize their nutritional requirements. It is critically important

  19. Comparison of three cell fixation methods for high content analysis assays utilizing quantum dots.

    Science.gov (United States)

    Williams, Y; Byrne, S; Bashir, M; Davies, A; Whelan, A; Gun'ko, Y; Kelleher, D; Volkov, Y

    2008-10-01

    Semiconductor nanoparticles or quantum dots are being increasingly utilized as fluorescent probes in cell biology both in live and fixed cell assays. Quantum dots possess an immense potential for use in multiplexing assays that can be run on high content screening analysers. Depending on the nature of the biological target under investigation, experiments are frequently required on cells retaining an intact cell membrane or also on those that have been fixed and permeabilized to expose intracellular antigens. Fixation of cell lines before or after the addition of quantum dots may affect their localization, emission properties and stability. Using a high content analysis platform we perform a quantitative comparative analysis of three common fixation techniques in two different cell lines exposed to carboxylic acid stabilized CdTe quantum dots. Our study demonstrates that in prefixed and permeabilized cells, quantum dots are readily internalized regardless of cell type, and their intracellular location is primarily determined by the properties of the quantum dots themselves. However, if the fixation procedures are preformed on live cells previously incubated with quantum dots, other important factors have to be considered. The choice of the fixative significantly influences the fluorescent characteristics of the quantum dots. Fixatives, regardless of their chemical nature, negatively affected quantum dots fluorescence intensity. Comparative analysis of gluteraldehyde, methanol and paraformaldehyde demonstrated that 2% paraformaldehyde was the fixative of choice. The presence of protein in the media did not significantly alter the quantum dot fluorescence. This study indicates that multiplexing assays utilizing quantum dots, despite being a cutting edge tool for high content cell imaging, still require careful consideration of the basic steps in biological sample processing.

  20. [Characteristics of anaerobic sequencing batch reactor for the treatment of high-solids-content waste].

    Science.gov (United States)

    Wang, Zhi-jun; Wang, Wei; Zhang, Xi-hui

    2006-06-01

    Based on the experiments of digestion of thermo-hydrolyzed sewage sludge in both mesophilic and thermophilic anaerobic sequencing batch reactors (ASBRs) with 20, 10, 7.5, 5d hydraulic retention time (HRT), operating characteristics of ASBR for treatment of high-solids-content waste were investigated. ASBR can efficiently accumulates suspended solids and keep high concentration solids, however there exists a "critical point" of ASBR, which means the maximum capability to accumulate suspended solids without negative effects on ASBR stability, and beyond which the performance deteriorates. Under steady condition, ASBR can sustains high solid retention time (SRT) and mean cell retention time (MCRT), the SRT and MCRT is 2.53 approximately 3.73 and 2.03 approximately 3.14 times of hydraulic retention time (HRT) when treating thermo-hydrolyzed sludge, respectively. Therefore, compared to traditional continuous-flow stirred tank reactor (CSTR), the efficiency of ASBR enhances about 7.13% approximately 34.68%.

  1. FBC desulfurization process using coal with low sulfur content, high oxidizing conditions and metamorphic limestones

    Directory of Open Access Journals (Sweden)

    S. R. Bragança

    2009-06-01

    Full Text Available A metamorphic limestone and a dolomite were employed as SO2 sorbents in the desulfurization of gas from coal combustion. The tests were performed in a fluidized bed reactor on a bench and pilot scale. Several parameters such as bed temperature, sorbent type, and sorbent particle size at different Ca/S molar ratios were analyzed. These parameters were evaluated for the combustion of coal with low-sulfur/high-ash content, experimental conditions of high air excess and high O2 level in fluidization air. Under these conditions, typical of furnaces, few published data can be found. In this work, a medium level of desulfurization efficiency (~60% for Ca/S = 2 was obtained.

  2. FBC desulfurization process using coal with low sulfur content, high oxidizing conditions and metamorphic limestones

    Energy Technology Data Exchange (ETDEWEB)

    Braganca, S.R.; Castellan, J.L. [Universidade Federal do Rio Grande do Sul, Porto Alegre (Brazil)

    2009-04-15

    A metamorphic limestone and a dolomite were employed as SO{sub 2} sorbents in the desulfurization of gas from coal combustion. The tests were performed in a fluidized bed reactor on a bench and pilot scale. Several parameters such as bed temperature, sorbent type, and sorbent particle size at different Ca/S molar ratios were analyzed. These parameters were evaluated for the combustion of coal with low-sulfur/high-ash content, experimental conditions of high air excess and high O{sub 2} level in fluidization air. Under these conditions, typical of furnaces, few published data can be found. In this work, a medium level of desulfurization efficiency (similar to 60%) for Ca/S = 2 was obtained.

  3. FBC desulfurization process using coal with low sulfur content, high oxidizing conditions and metamorphic limestones

    Energy Technology Data Exchange (ETDEWEB)

    S.R. Braganca; J.L. Castellan [Federal University of Rio Grande do Sul, Porto Alegre (Brazil)

    2009-07-01

    A metamorphic limestone and a dolomite were employed as SO{sub 2} sorbents in the desulfurization of gas from coal combustion. The tests were performed in a fluidised bed reactor on a bench and pilot scale. Several parameters such as bed temperature, sorbent type, and sorbent particle size at different Ca/S molar ratios were analyzed. These parameters were evaluated for the combustion of coal with low-sulfur/high-ash content, experimental conditions of high air excess and high O{sub 2} level in fluidization air. Under these conditions, typical of furnaces, few published data can be found. In this work, a medium level of desulfurization efficiency (about 60%) for Ca/S = 2 was obtained. 25 refs., 5 figs.

  4. High-content analysis screening for cell cycle regulators using arrayed synthetic crRNA libraries.

    Science.gov (United States)

    Strezoska, Žaklina; Perkett, Matthew R; Chou, Eldon T; Maksimova, Elena; Anderson, Emily M; McClelland, Shawn; Kelley, Melissa L; Vermeulen, Annaleen; Smith, Anja van Brabant

    2017-06-10

    The CRISPR-Cas9 system has been utilized for large-scale, loss-of-function screens mainly using lentiviral pooled formats and cell-survival phenotypic assays. Screening in an arrayed format expands the types of phenotypic readouts that can be used to now include high-content, morphology-based assays, and with the recent availability of synthetic crRNA libraries, new studies are emerging. Here, we use a cell cycle reporter cell line to perform an arrayed, synthetic crRNA:tracrRNA screen targeting 169 genes (>600 crRNAs) and used high content analysis (HCA) to identify genes that regulate the cell cycle. Seven parameters were used to classify cells into cell cycle categories and multiple parameters were combined using a new analysis technique to identify hits. Comprehensive hit follow-up experiments included target gene expression analysis, confirmation of DNA insertions/deletions, and validation with orthogonal reagents. Our results show that most hits had three or more independent crRNAs per gene that demonstrated a phenotype with consistent individual parameters, indicating that our screen produced high-confidence hits with low off-target effects and allowed us to identify hits with more subtle phenotypes. The results of our screen demonstrate the power of using arrayed, synthetic crRNAs for functional phenotypic screening using multiparameter HCA assays. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

  5. Tenside-free preparation of nanogels with high functional β-cyclodextrin content.

    Science.gov (United States)

    Kettel, Markus J; Hildebrandt, Haika; Schaefer, Karola; Moeller, Martin; Groll, Juergen

    2012-09-25

    We present the preparation of ultrafine (R(h), 50 -150 nm) nanogels through tenside-free condensation of reactive prepolymers with β-cyclodextrin (β-CD) in water. These nanogels possess a maximum content of 60 wt % functional β-CD that can form inclusion complexes as demonstrated by dye sorption with phenolphthalein. Aside of this extremely high uptake capacity to hydrophobic molecules, the nanogels also show good adhesion to surfaces in homogeneous distribution with size of R(h) of 25 nm under dry conditions.

  6. Process Modules for GeSn Nanoelectronics with high Sn-contents

    OpenAIRE

    Schulte-Braucks, C; Glass, S; Hofmann, E; Stange, D; Von Den Driesch, N; Hartmann, JM; Ikonic, Z; Zhao, GT; Buca, D.; Mantl, S

    2017-01-01

    This paper systematically studies GeSn n-FETs, from individual process modules to a complete device. High-k gate stacks and NiGeSn metallic contacts for source and drain are characterized in independent experiments. To study both direct and indirect bandgap semiconductors, a range of 0 at.% to 14.5 at.% Sn-content GeSn alloys are investigated. Special emphasis is placed on capacitance-voltage (C-V) characteristics and Schottky-barrier optimization. GeSn n-FET devices are presented including t...

  7. Desulphurisation of high moisture content fuel-gases derived from low-rank coals

    Energy Technology Data Exchange (ETDEWEB)

    Hodges, S.; Anderson, B. [HRL Technology, Mulgrave, Vic. (Australia); Abbasian, J.; Slimane, R.B. [Inst. of Gas Technology, Des Plaines, IL (United States)

    1999-07-01

    Regenerable sulphur sorbent materials have been developed specifically for fluidised-bed desulphurisation of high moisture content fuel-gases derived from the gasification of low-rank coals. Selection of the most appropriate sorbents was based on thermodynamic limitations, strength/attrition resistance, reactivity and sulphur capacity. Pilot-scale tests showed that sorbents based on iron and copper were able to reduce the level of H{sub 2}S in the fuel-gas (up to 2.5 MPa, 350 C) from about 3000 ppmv to less than 100 ppmv. (orig.)

  8. Optimal Dearsenification Parameters of Gold Sulfide Concentrate with a High As Content

    Institute of Scientific and Technical Information of China (English)

    Yaozhong LAN; Rose W. Smith

    2005-01-01

    The gold sulfide concentrate with a high As content in Liangshan District, Sichuan Province, China, is a potentially important resource. This paper describes experiments of dearsenification of gold concentrate in a weakly reduced atmosphere in a rotary pipe furnace. The results showed that the optimal parameters were a temperature range of 650~700℃, 15%~16% CO2 of gas and a reaction time of 30~40 min. The removal rate of arsenic and sulfur was over 95% and 25%~28%, respectively. With further oxidization and roasting, residue sulfur in the roasted materials was dropped to below 4%, and the cyanide leaching recovery of gold was over 92%.

  9. Non-metallic inclusions structure dimension in high quality steel with medium carbon contents

    Directory of Open Access Journals (Sweden)

    T. Lipiński

    2009-07-01

    Full Text Available The experimental material consisted of semi-finished products of high-grade, medium-carbon structural steel. The production process involved two melting technologies: steel melting in a 140-ton basic arc furnace with desulfurization and argon refining variants, and in a 100-ton oxygen converter. Billet samples were collected to analyze the content of non-metallic inclusions with the use of an optical microscope and a video inspection microscope. The results were processed and presented in graphic form.

  10. Identification of Novel Macropinocytosing Human Antibodies by Phage Display and High-Content Analysis.

    Science.gov (United States)

    Ha, K D; Bidlingmaier, S M; Su, Y; Lee, N-K; Liu, B

    2017-01-01

    Internalizing antibodies have great potential for the development of targeted therapeutics. Antibodies that internalize via the macropinocytosis pathway are particularly promising since macropinocytosis is capable of mediating rapid, bulk uptake and is selectively upregulated in many cancers. We hereby describe a method for identifying antibodies that internalize via macropinocytosis by screening phage-displayed single-chain antibody selection outputs with an automated fluorescent microscopy-based high-content analysis platform. Furthermore, this method can be similarly applied to other endocytic pathways if other fluorescent, pathway-specific, soluble markers are available. © 2017 Elsevier Inc. All rights reserved.

  11. Cellular Telephone

    Institute of Scientific and Technical Information of China (English)

    杨周

    1996-01-01

    Cellular phones, used in automobiles, airliners, and passenger trains, are basically low-power radiotelephones. Calls go through radio transmitters that are located within small geographical units called cells. Because each cell’s signals are too weak to interfere with those of other cells operating on the same fre-

  12. The endocytic receptor megalin binds the iron transporting neutrophil-gelatinase-associated lipocalin with high affinity and mediates its cellular uptake

    DEFF Research Database (Denmark)

    Hvidberg, Vibeke; Jacobsen, Christian; Strong, Roland K

    2005-01-01

    in delivering iron to cells during formation of the tubular epithelial cells of the primordial kidney. No cellular receptor for NGAL has been described. We show here that megalin, a member of the low-density lipoprotein receptor family expressed in polarized epithelia, binds NGAL with high affinity, as shown...

  13. Highly Ordered 1D Fullerene Crystals for Concurrent Control of Macroscopic Cellular Orientation and Differentiation toward Large-Scale Tissue Engineering.

    Science.gov (United States)

    Minami, Kosuke; Kasuya, Yuki; Yamazaki, Tomohiko; Ji, Qingmin; Nakanishi, Waka; Hill, Jonathan P; Sakai, Hideki; Ariga, Katsuhiko

    2015-07-15

    A highly aligned 1D fullerene whisker (FW) scaffold in a centimeter area is fabricated by interfacial alignment. The resulting aligned FW scaffold enables concurrent control over cellular orientation and differentiation to muscle cells. This aligned FW scaffold is made by a facile method, and hence the substrate is a promising alternative to other cell scaffolds for tissue engineering.

  14. Experiences of High School Students about the Predictors of Tobacco Use: a Directed Qualitative Content Analysis

    Directory of Open Access Journals (Sweden)

    Mahmoud Ghasemi

    2015-12-01

    Full Text Available Background and Objectives: Tobacco use is one of the most important risk factors that increases the burden of diseases worldwide. Based on the increasing speed of tobacco use, the aim of the present study was to explain the experiences of high school students about the determiners of use and non-use of tobacco (cigarettes and hookah based on the theory of protection motivation. Materials and Methods: The present study is a qualitative study based on content analysis that has been carried out for five months from 22, November of 2014 to 20, April of 2015 on male high schools in Noshahr. Data were collected in the form of semi-structured interviews from 21 male high school students of whom 7 smoked cigarettes, 7 used hookah and 7 of them did not use any type of tobacco. Data analysis was carried out through the use of directed qualitative content analysis. Results: Data analysis led to the extraction of 99 primary codes that were categorized into 9 predetermined levels of protection motivation theory including perceived sensitivity, perceived intensity, fear, perceived self-efficacy, response expense, efficiency of the perceived answer, external perceived reward, internal perceived reward, protection motivation. The findings of the study showed that the most important predictors for the use of tobacco were the structures of response expense and high perceived rewards and the most important predictors for non-use of tobacco were perceived sensitivity, perceived intensity and high self-efficacy of students. Conclusions: the findings of the present study showed that the pressure from peers, being present in a group using tobacco and the absence of alternative recreational activities are among the most important factors of using tobacco. So, it is suggested that planners of the health department take the comprehensive interventions to improve effective individual and environmental factors of using tobacco so that they could reduce smoking cigarettes

  15. Biochemistry in the High School Textbook: A Distance Between Content And Students Reality?

    Directory of Open Access Journals (Sweden)

    A. Simões-Pires

    2012-05-01

    Full Text Available Several  contents  have  been  taught  in  a  decontextualized  way  within  the  area  of biology,  one  of  these  contents  is  Biochemistry.  Biochemistry  is  one  of  the  most present subjects in the student’s daily lives, because it can explain a huge number of  processes  that  happens  in  their  routine.  The  textbook  is  the  main  tool  in  the Brazilian  public  high  schools  classrooms,  so  a  huge  number  of  programs  to improve the textbooks were created by the government, like the National Program of the Textbook or PNLD( translated from the Portuguese ‘’Programa Nacional do Livro Didático’’. But despite the efforts to improve the textbook, their quality is still questionable, for such reasons like the distance between contents showed in the textbook and student’s reality. The main objective of this work is to evaluate how the  high  school  textbooks  show  the  content  related  to  biochemistry  and  how  this content is contextualized in the student’s reality.  To achieve this goal we analyzed four  textbooks  that  are  most  frequently  used  in  the  schools  today  and  evaluated parameters  related  with  contextualization  between  the  biochemistry  content  and students  daily  lives  situations.  The  analysis  was  done  on  a  qualitative  way.  We observed  a  lack  of  daily  examples  in  the  biochemical  contents,  an  excess  of scientific terms without clear explanations and also a huge presence of conceptual errors  in  all  four  textbooks.  The  data  showed  here  leads  us  to  conclude  that  the science’s textbooks analyzed in this study are inappropriate to teach biochemistry in an interesting way for the students.

  16. High-throughput investigation of endothelial-to-mesenchymal transformation (EndMT) with combinatorial cellular microarrays.

    Science.gov (United States)

    Wang, Zongjie; Calpe, Blaise; Zerdani, Jalil; Lee, Youngsang; Oh, Jonghyun; Bae, Hojae; Khademhosseini, Ali; Kim, Keekyoung

    2016-07-01

    In the developing heart, a specific subset of endocardium undergoes an endothelial-to-mesenchymal transformation (EndMT) thus forming nascent valve leaflets. Extracellular matrix (ECM) proteins and growth factors (GFs) play important roles in regulating EndMT but the combinatorial effect of GFs with ECM proteins is less well understood. Here we use microscale engineering techniques to create single, binary, and tertiary component microenvironments to investigate the combinatorial effects of ECM proteins and GFs on the attachment and transformation of adult ovine mitral valve endothelial cells to a mesenchymal phenotype. With the combinatorial microenvironment microarrays, we utilized 60 different combinations of ECM proteins (Fibronectin, Collagen I, II, IV, Laminin) and GFs (TGF-β1, bFGF, VEGF) and were able to identify new microenvironmental conditions capable of modulating EndMT in MVECs. Experimental results indicated that TGF-β1 significantly upregulated the EndMT while either bFGF or VEGF downregulated EndMT process markedly. Also, ECM proteins could influence both the attachment of MVECs and the response of MVECs to GFs. In terms of attachment, fibronectin is significantly better for the adhesion of MVECs among the five tested proteins. Overall collagen IV and fibronectin appeared to play important roles in promoting EndMT process. Great consistency between macroscale and microarrayed experiments and present studies demonstrates that high-throughput cellular microarrays are a promising approach to study the regulation of EndMT in valvular endothelium. Biotechnol. Bioeng. 2016;113: 1403-1412. © 2015 Wiley Periodicals, Inc.

  17. High-content screening of drug-induced cardiotoxicity using quantitative single cell imaging cytometry on microfluidic device.

    Science.gov (United States)

    Kim, Min Jung; Lee, Su Chul; Pal, Sukdeb; Han, Eunyoung; Song, Joon Myong

    2011-01-07

    Drug-induced cardiotoxicity or cytotoxicity followed by cell death in cardiac muscle is one of the major concerns in drug development. Herein, we report a high-content quantitative multicolor single cell imaging tool for automatic screening of drug-induced cardiotoxicity in an intact cell. A tunable multicolor imaging system coupled with a miniaturized sample platform was destined to elucidate drug-induced cardiotoxicity via simultaneous quantitative monitoring of intracellular sodium ion concentration, potassium ion channel permeability and apoptosis/necrosis in H9c2(2-1) cell line. Cells were treated with cisapride (a human ether-à-go-go-related gene (hERG) channel blocker), digoxin (Na(+)/K(+)-pump blocker), camptothecin (anticancer agent) and a newly synthesized anti-cancer drug candidate (SH-03). Decrease in potassium channel permeability in cisapride-treated cells indicated that it can also inhibit the trafficking of the hERG channel. Digoxin treatment resulted in an increase of intracellular [Na(+)]. However, it did not affect potassium channel permeability. Camptothecin and SH-03 did not show any cytotoxic effect at normal use (≤300 nM and 10 μM, respectively). This result clearly indicates the potential of SH-03 as a new anticancer drug candidate. The developed method was also used to correlate the cell death pathway with alterations in intracellular [Na(+)]. The developed protocol can directly depict and quantitate targeted cellular responses, subsequently enabling an automated, easy to operate tool that is applicable to drug-induced cytotoxicity monitoring with special reference to next generation drug discovery screening. This multicolor imaging based system has great potential as a complementary system to the conventional patch clamp technique and flow cytometric measurement for the screening of drug cardiotoxicity.

  18. Behavioral and cellular consequences of high-electrode count Utah Arrays chronically implanted in rat sciatic nerve

    Science.gov (United States)

    Wark, H. A. C.; Mathews, K. S.; Normann, R. A.; Fernandez, E.

    2014-08-01

    Objective. Before peripheral nerve electrodes can be used for the restoration of sensory and motor functions in patients with neurological disorders, the behavioral and histological consequences of these devices must be investigated. These indices of biocompatibility can be defined in terms of desired functional outcomes; for example, a device may be considered for use as a therapeutic intervention if the implanted subject retains functional neurons post-implantation even in the presence of a foreign body response. The consequences of an indwelling device may remain localized to cellular responses at the device-tissue interface, such as fibrotic encapsulation of the device, or they may affect the animal more globally, such as impacting behavioral or sensorimotor functions. The objective of this study was to investigate the overall consequences of implantation of high-electrode count intrafascicular peripheral nerve arrays, High Density Utah Slanted Electrode Arrays (HD-USEAs; 25 electrodes mm-2). Approach. HD-USEAs were implanted in rat sciatic nerves for one and two month periods. We monitored wheel running, noxious sensory paw withdrawal reflexes, footprints, nerve morphology and macrophage presence at the tissue-device interface. In addition, we used a novel approach to contain the arrays in actively behaving animals that consisted of an organic nerve wrap. A total of 500 electrodes were implanted across all ten animals. Main results. The results demonstrated that chronic implantation (⩽8 weeks) of HD-USEAs into peripheral nerves can evoke behavioral deficits that recover over time. Morphology of the nerve distal to the implantation site showed variable signs of nerve fiber degeneration and regeneration. Cytology adjacent to the device-tissue interface also showed a variable response, with some electrodes having many macrophages surrounding the electrodes, while other electrodes had few or no macrophages present. This variability was also seen along the length

  19. Fluorescence correlation spectroscopy as tool for high-content-screening in yeast (HCS-FCS)

    Science.gov (United States)

    Wood, Christopher; Huff, Joseph; Marshall, Will; Yu, Elden Qingfeng; Unruh, Jay; Slaughter, Brian; Wiegraebe, Winfried

    2011-03-01

    To measure protein interactions, diffusion properties, and local concentrations in single cells, Fluorescence Correlation Spectroscopy (FCS) is a well-established and widely accepted method. However, measurements can take a long time and are laborious. Therefore investigations are typically limited to tens or a few hundred cells. We developed an automated system to overcome these limitations and make FCS available for High Content Screening (HCS). We acquired data in an auto-correlation screen of more than 4000 of the 6000 proteins of the yeast Saccharomyces cerevisiae, tagged with eGFP and expanded the HCS to use cross-correlation between eGFP and mCherry tagged proteins to screen for molecular interactions. We performed all high-content FCS screens (HCS-FCS) in a 96 well plate format. The system is based on an extended Carl Zeiss fluorescence correlation spectrometer ConfoCor 3 attached to a confocal microscope LSM 510. We developed image-processing software to control these hardware components. The confocal microscope obtained overview images and we developed an algorithm to search for and detect single cells. At each cell, we positioned a laser beam at a well-defined point and recorded the fluctuation signal. We used automatic scoring of the signal for quality control. All data was stored and organized in a database based on the open source Open Microscopy Environment (OME) platform. To analyze the data we used the image processing language IDL and the open source statistical software package R.

  20. BioSig3D: High Content Screening of Three-Dimensional Cell Culture Models.

    Directory of Open Access Journals (Sweden)

    Cemal Cagatay Bilgin

    Full Text Available BioSig3D is a computational platform for high-content screening of three-dimensional (3D cell culture models that are imaged in full 3D volume. It provides an end-to-end solution for designing high content screening assays, based on colony organization that is derived from segmentation of nuclei in each colony. BioSig3D also enables visualization of raw and processed 3D volumetric data for quality control, and integrates advanced bioinformatics analysis. The system consists of multiple computational and annotation modules that are coupled together with a strong use of controlled vocabularies to reduce ambiguities between different users. It is a web-based system that allows users to: design an experiment by defining experimental variables, upload a large set of volumetric images into the system, analyze and visualize the dataset, and either display computed indices as a heatmap, or phenotypic subtypes for heterogeneity analysis, or download computed indices for statistical analysis or integrative biology. BioSig3D has been used to profile baseline colony formations with two experiments: (i morphogenesis of a panel of human mammary epithelial cell lines (HMEC, and (ii heterogeneity in colony formation using an immortalized non-transformed cell line. These experiments reveal intrinsic growth properties of well-characterized cell lines that are routinely used for biological studies. BioSig3D is being released with seed datasets and video-based documentation.

  1. Human pluripotent stem cells on artificial microenvironments: a high content perspective.

    Directory of Open Access Journals (Sweden)

    Priyalakshmi eViswanathan

    2014-07-01

    Full Text Available Self-renewing stem cell populations are increasingly considered as resources for cell therapy and tools for drug discovery. Human pluripotent stem (hPS cells in particular offer a virtually unlimited reservoir of homogeneous cells and can be differentiated towards diverse lineages. Many diseases show impairment in self-renewal or differentiation, abnormal lineage choice or other aberrant cell behavior in response to chemical or physical cues. To investigate these responses, there is a growing interest in the development of specific assays using hPS cells artificial microenvironments and high content analysis. Several hurdles need to be overcome that can be grouped in: (i availability of robust, homogeneous and consistent cell populations as a starting point; (ii appropriate understanding and use of chemical and physical microenvironments; (iii development of assays that dissect the complexity of cell populations in tissues while mirroring specific aspects of their behavior. Here we review recent progress in the culture of hPS cells and we detail the importance of the environment surrounding the cells with a focus on synthetic material and suitable high content analysis approaches. The technologies described if properly combined have the potential to create a paradigm shift in the way diseases are modelled and drug discovery is performed.

  2. A Liposomal Formulation Able to Incorporate a High Content of Paclitaxel and Exert Promising Anticancer Effect

    Directory of Open Access Journals (Sweden)

    Pei Kan

    2011-01-01

    Full Text Available A liposome formulation for paclitaxel was developed in this study. The liposomes, composed of naturally unsaturated and hydrogenated phosphatidylcholines, with significant phase transition temperature difference, were prepared and characterized. The liposomes exhibited a high content of paclitaxel, which was incorporated within the segregated microdomains coexisting on phospholipid bilayer of liposomes. As much as 15% paclitaxel to phospholipid molar ratio were attained without precipitates observed during preparation. In addition, the liposomes remained stable in liquid form at 4∘C for at least 6 months. The special composition of liposomal membrane which could reduce paclitaxel aggregation could account for such a capacity and stability. The cytotoxicity of prepared paclitaxel liposomes on the colon cancer C-26 cell culture was comparable to Taxol. Acute toxicity test revealed that LD50 for intravenous bolus injection in mice exceeded by 40 mg/kg. In antitumor efficacy study, the prepared liposomal paclitaxel demonstrated the increase in the efficacy against human cancer in animal model. Taken together, the novel formulated liposomes can incorporate high content of paclitaxel, remaining stable for long-term storage. These animal data also demonstrate that the liposomal paclitaxel is promising for further clinical use.

  3. High-content analysis of single cells directly assembled on CMOS sensor based on color imaging.

    Science.gov (United States)

    Tanaka, Tsuyoshi; Saeki, Tatsuya; Sunaga, Yoshihiko; Matsunaga, Tadashi

    2010-12-15

    A complementary metal oxide semiconductor (CMOS) image sensor was applied to high-content analysis of single cells which were assembled closely or directly onto the CMOS sensor surface. The direct assembling of cell groups on CMOS sensor surface allows large-field (6.66 mm×5.32 mm in entire active area of CMOS sensor) imaging within a second. Trypan blue-stained and non-stained cells in the same field area on the CMOS sensor were successfully distinguished as white- and blue-colored images under white LED light irradiation. Furthermore, the chemiluminescent signals of each cell were successfully visualized as blue-colored images on CMOS sensor only when HeLa cells were placed directly on the micro-lens array of the CMOS sensor. Our proposed approach will be a promising technique for real-time and high-content analysis of single cells in a large-field area based on color imaging.

  4. Development of high-content assays for kidney progenitor cell expansion in transgenic zebrafish.

    Science.gov (United States)

    Sanker, Subramaniam; Cirio, Maria Cecilia; Vollmer, Laura L; Goldberg, Natasha D; McDermott, Lee A; Hukriede, Neil A; Vogt, Andreas

    2013-12-01

    Reactivation of genes normally expressed during organogenesis is a characteristic of kidney regeneration. Enhancing this reactivation could potentially be a therapeutic target to augment kidney regeneration. The inductive events that drive kidney organogenesis in zebrafish are similar to the initial steps in mammalian kidney organogenesis. Therefore, quantifying embryonic signals that drive zebrafish kidney development is an attractive strategy for the discovery of potential novel therapeutic modalities that accelerate kidney regeneration. The Lim1 homeobox protein, Lhx1, is a marker of kidney development that is also expressed in the regenerating kidneys after injury. Using a fluorescent Lhx1a-EGFP transgene whose phenotype faithfully recapitulates that of the endogenous protein, we developed a high-content assay for Lhx1a-EGFP expression in transgenic zebrafish embryos employing an artificial intelligence-based image analysis method termed cognition network technology (CNT). Implementation of the CNT assay on high-content readers enabled automated real-time in vivo time-course, dose-response, and variability studies in the developing embryo. The Lhx1a assay was complemented with a kidney-specific secondary CNT assay that enables direct measurements of the embryonic renal tubule cell population. The integration of fluorescent transgenic zebrafish embryos with automated imaging and artificial intelligence-based image analysis provides an in vivo analysis system for structure-activity relationship studies and de novo discovery of novel agents that augment innate regenerative processes.

  5. Two-step in situ biodiesel production from microalgae with high free fatty acid content.

    Science.gov (United States)

    Dong, Tao; Wang, Jun; Miao, Chao; Zheng, Yubin; Chen, Shulin

    2013-05-01

    The yield of fatty acid methyl ester (FAME) from microalgae biomass is generally low via traditional extraction-conversion route due to the deficient solvent extraction. In this study a two-step in situ process was investigated to obtain a high FAME yield from microalgae biomass that had high free fatty acids (FFA) content. This was accomplished with a pre-esterification process using heterogeneous catalyst to reduce FFA content prior to the base-catalyzed transesterification. The two-step in situ process resulted in a total FAME recovery up to 94.87±0.86%, which was much higher than that obtained by a one-step acid or base catalytic in situ process. The heterogeneous catalyst, Amberlyst-15, could be used for 8 cycles without significant loss in activity. This process have the potential to reduce the production cost of microalgae-derived FAME and be more environmental compatible due to the higher FAME yield with reduced catalyst consumption.

  6. BioSig3D: High Content Screening of Three-Dimensional Cell Culture Models.

    Science.gov (United States)

    Bilgin, Cemal Cagatay; Fontenay, Gerald; Cheng, Qingsu; Chang, Hang; Han, Ju; Parvin, Bahram

    2016-01-01

    BioSig3D is a computational platform for high-content screening of three-dimensional (3D) cell culture models that are imaged in full 3D volume. It provides an end-to-end solution for designing high content screening assays, based on colony organization that is derived from segmentation of nuclei in each colony. BioSig3D also enables visualization of raw and processed 3D volumetric data for quality control, and integrates advanced bioinformatics analysis. The system consists of multiple computational and annotation modules that are coupled together with a strong use of controlled vocabularies to reduce ambiguities between different users. It is a web-based system that allows users to: design an experiment by defining experimental variables, upload a large set of volumetric images into the system, analyze and visualize the dataset, and either display computed indices as a heatmap, or phenotypic subtypes for heterogeneity analysis, or download computed indices for statistical analysis or integrative biology. BioSig3D has been used to profile baseline colony formations with two experiments: (i) morphogenesis of a panel of human mammary epithelial cell lines (HMEC), and (ii) heterogeneity in colony formation using an immortalized non-transformed cell line. These experiments reveal intrinsic growth properties of well-characterized cell lines that are routinely used for biological studies. BioSig3D is being released with seed datasets and video-based documentation.

  7. The Gray Institute ‘open’ high-content, fluorescence lifetime microscopes

    Science.gov (United States)

    BARBER, PR; TULLIS, IDC; PIERCE, GP; NEWMAN, RG; PRENTICE, J; ROWLEY, MI; MATTHEWS, DR; AMEER-BEG, SM; VOJNOVIC, B

    2013-01-01

    Summary We describe a microscopy design methodology and details of microscopes built to this ‘open’ design approach. These demonstrate the first implementation of time-domain fluorescence microscopy in a flexible automated platform with the ability to ease the transition of this and other advanced microscopy techniques from development to use in routine biology applications. This approach allows easy expansion and modification of the platform capabilities, as it moves away from the use of a commercial, monolithic, microscope body to small, commercial off-the-shelf and custom made modular components. Drawings and diagrams of our microscopes have been made available under an open license for noncommercial use at http://users.ox.ac.uk/~atdgroup. Several automated high-content fluorescence microscope implementations have been constructed with this design framework and optimized for specific applications with multiwell plates and tissue microarrays. In particular, three platforms incorporate time-domain FLIM via time-correlated single photon counting in an automated fashion. We also present data from experiments performed on these platforms highlighting their automated wide-field and laser scanning capabilities designed for high-content microscopy. Devices using these designs also form radiation-beam ‘end-stations’ at Oxford and Surrey Universities, showing the versatility and extendibility of this approach. PMID:23772985

  8. Corrosion Behavior of the Stressed Sensitized Austenitic Stainless Steels of High Nitrogen Content in Seawater

    Directory of Open Access Journals (Sweden)

    A. Almubarak

    2013-01-01

    Full Text Available The purpose of this paper is to study the effect of high nitrogen content on corrosion behavior of austenitic stainless steels in seawater under severe conditions such as tensile stresses and existence of sensitization in the structure. A constant tensile stress has been applied to sensitized specimens types 304, 316L, 304LN, 304NH, and 316NH stainless steels. Microstructure investigation revealed various degrees of stress corrosion cracking. SCC was severe in type 304, moderate in types 316L and 304LN, and very slight in types 304NH and 316NH. The electrochemical polarization curves showed an obvious second current peak for the sensitized alloys which indicated the existence of second phase in the structure and the presence of intergranular stress corrosion cracking. EPR test provided a rapid and efficient nondestructive testing method for showing passivity, degree of sensitization and determining IGSCC for stainless steels in seawater. A significant conclusion was obtained that austenitic stainless steels of high nitrogen content corrode at a much slower rate increase pitting resistance and offer an excellent resistance to stress corrosion cracking in seawater.

  9. Kenaf Powder Filled Recycled High Density Polyethylene/Natural Rubber Biocomposites: The Effect of Filler Content

    Directory of Open Access Journals (Sweden)

    Xuan Viet Cao

    2012-09-01

    Full Text Available The performance of kenaf powder (KP as filler for recycled high density polyethylene (rHDPE/natural rubber (NR thermoplastic elastomer (TPE composites was investigated. The composites with different filler loading were prepared in a Haake internal mixer. Increasing KP loading in rHDPE/NR/KP biocomposites reduced the tensile strength, elongation at break but increased the stabilization torque and the tensile modulus. SEM study of fracture surface indicated that fibrillation of rHDPE was reduced and detachment of kenaf powder from polymer matrix was present particularly at high filler loading. These observations were responsible for the deterioration of tensile strength and elongation at break of rHDPE/NR/KP biocomposites. Water absorption study also showed that the water absorption of these biocomposites increased with increasing KP content.

  10. High-Content Analysis of CRISPR-Cas9 Gene-Edited Human Embryonic Stem Cells

    Directory of Open Access Journals (Sweden)

    Jared Carlson-Stevermer

    2016-01-01

    Full Text Available CRISPR-Cas9 gene editing of human cells and tissues holds much promise to advance medicine and biology, but standard editing methods require weeks to months of reagent preparation and selection where much or all of the initial edited samples are destroyed during analysis. ArrayEdit, a simple approach utilizing surface-modified multiwell plates containing one-pot transcribed single-guide RNAs, separates thousands of edited cell populations for automated, live, high-content imaging and analysis. The approach lowers the time and cost of gene editing and produces edited human embryonic stem cells at high efficiencies. Edited genes can be expressed in both pluripotent stem cells and differentiated cells. This preclinical platform adds important capabilities to observe editing and selection in situ within complex structures generated by human cells, ultimately enabling optical and other molecular perturbations in the editing workflow that could refine the specificity and versatility of gene editing.

  11. Comparison of multivariate data analysis strategies for high-content screening.

    Science.gov (United States)

    Kümmel, Anne; Selzer, Paul; Beibel, Martin; Gubler, Hanspeter; Parker, Christian N; Gabriel, Daniela

    2011-03-01

    High-content screening (HCS) is increasingly used in biomedical research generating multivariate, single-cell data sets. Before scoring a treatment, the complex data sets are processed (e.g., normalized, reduced to a lower dimensionality) to help extract valuable information. However, there has been no published comparison of the performance of these methods. This study comparatively evaluates unbiased approaches to reduce dimensionality as well as to summarize cell populations. To evaluate these different data-processing strategies, the prediction accuracies and the Z' factors of control compounds of a HCS cell cycle data set were monitored. As expected, dimension reduction led to a lower degree of discrimination between control samples. A high degree of classification accuracy was achieved when the cell population was summarized on well level using percentile values. As a conclusion, the generic data analysis pipeline described here enables a systematic review of alternative strategies to analyze multiparametric results from biological systems.

  12. Selective-plane illumination microscopy for high-content volumetric biological imaging

    Science.gov (United States)

    McGorty, Ryan; Huang, Bo

    2016-03-01

    Light-sheet microscopy, also named selective-plane illumination microscopy, enables optical sectioning with minimal light delivered to the sample. Therefore, it allows one to gather volumetric datasets of developing embryos and other light-sensitive samples over extended times. We have configured a light-sheet microscope that, unlike most previous designs, can image samples in formats compatible with high-content imaging. Our microscope can be used with multi-well plates or with microfluidic devices. In designing our optical system to accommodate these types of sample holders we encounter large optical aberrations. We counter these aberrations with both static optical components in the imaging path and with adaptive optics. Potential applications of this microscope include studying the development of a large number of embryos in parallel and over long times with subcellular resolution and doing high-throughput screens on organisms or cells where volumetric data is necessary.

  13. Performance of a commercial inoculum for the aerobic biodegradation of a high fat content dairy wastewater.

    Science.gov (United States)

    Loperena, Lyliam; Ferrari, Mario Daniel; Saravia, Verónica; Murro, Daiman; Lima, Cynthia; Ferrando, Lucía; Fernández, Ana; Lareo, Claudia

    2007-03-01

    The effectiveness of a commercial inoculum for degrading a dairy wastewater with high fat content was evaluated, and compared with an activated sludge inoculum from a dairy wastewater treatment pond. Both inocula reached similar chemical oxygen demand removal in batch experiments. The population dynamics was also studied by determining heterotrophic counts. Predominant microorganisms were differentiated by colony morphology and genomic fingerprinting (BOX-PCR) analysis. The higher population diversity and the wider range of CO2 production rate observed in batch reactors inoculated with activated-sludge, indicated that microorganisms from this inoculum were well adapted and may have had synergic activity for the degradation of the dairy effluent. When the bioreactor was operated with the commercial inoculum in continuous mode, according to its microbial growth kinetics, other microorganisms became predominant. These results showed that inoculated microorganisms did not persist in the open system and periodic addition of microorganisms may be needed to achieve a high performance treatment.

  14. Construction and validation of two metagenomic DNA libraries from Cerrado soil with high clay content.

    Science.gov (United States)

    de Castro, Alinne Pereira; Quirino, Betania Ferraz; Allen, Heather; Williamson, Lynn L; Handelsman, Jo; Krüger, Ricardo Henrique

    2011-11-01

    A challenge of metagenomic studies is in the extraction and purification of DNA from environmental samples. The soils of the Cerrado region of Brazil present several technical difficulties to DNA extraction: high clay content (>55% w/w), low pH (4.7) and high iron levels (146 ppm). Here we describe for the first time the efficient recovery and purification of microbial DNA associated with these unusual soil characteristics and the construction and validation of two metagenomic libraries: a 150,000 clones library with insert size of approximately 8 kb and a 65,000 clones library with insert size of approximately 35 kb. The construction of these metagenomic libraries will allow the biotechnological exploitation of the microbial community present in the soil from this endangered biome.

  15. High-Content Analysis of CRISPR-Cas9 Gene-Edited Human Embryonic Stem Cells.

    Science.gov (United States)

    Carlson-Stevermer, Jared; Goedland, Madelyn; Steyer, Benjamin; Movaghar, Arezoo; Lou, Meng; Kohlenberg, Lucille; Prestil, Ryan; Saha, Krishanu

    2016-01-12

    CRISPR-Cas9 gene editing of human cells and tissues holds much promise to advance medicine and biology, but standard editing methods require weeks to months of reagent preparation and selection where much or all of the initial edited samples are destroyed during analysis. ArrayEdit, a simple approach utilizing surface-modified multiwell plates containing one-pot transcribed single-guide RNAs, separates thousands of edited cell populations for automated, live, high-content imaging and analysis. The approach lowers the time and cost of gene editing and produces edited human embryonic stem cells at high efficiencies. Edited genes can be expressed in both pluripotent stem cells and differentiated cells. This preclinical platform adds important capabilities to observe editing and selection in situ within complex structures generated by human cells, ultimately enabling optical and other molecular perturbations in the editing workflow that could refine the specificity and versatility of gene editing.

  16. Salvinorin A content in legal high products of Salvia divinorum sold in Mexico.

    Science.gov (United States)

    Hernández-Bello, Rafael; García-Rodríguez, Rosa Virginia; García-Sosa, Karlina; Peña-Rodríguez, Luis Manuel; Vázquez-Hernández, Maribel; Ramos-Morales, Fernando Rafael; Corcoran, Olivia; Sánchez-Medina, Alberto

    2015-04-01

    Salvia divinorum (Lamiaceae) is a herb native to Mexico where it is used by Mazatec shamans for spiritual and divination purposes. S. divinorum products are easily available to consumers and are used worldwide as legal highs because of the hallucinogenic effects caused mainly by salvinorin A. Highly popular videos and websites on the internet depicting the use of S. divinorum products have contributed to an increase in their consumption. Recent reports have highlighted the potential of these products to induce psychosis in consumers. In Mexico, dried leaf extracts of S. divinorum are sold in different strengths, claiming to correlate with increasing amounts of salvinorin A. In order to determine the variability of salvinorin A content between brands and to investigate possible correlation between brand strengths, this study sought to quantify salvinorin A in commercial products available in Mexico using an HPLC method. The HPLC analytical method showed a correlation coefficient R(2)>0.99, with LOD of 0.44 μg/mL and LOQ of 1.34 μg/mL. The retention time for salvinorin A was 23.09±0.95 min and the measured concentrations ranged between 8.32±0.65 and 56.52±3.77 mg/g dried leaf. The results for brand c did not show an agreement between the declared and the calculated amount of salvinorin A. Additionally, the emergence in Mexico of high strength salvia products (100×), the lack of regulation and the observed variability of salvinorin A content between brands of commercial legal highs products of S. divinorum could result in a health problem for consumers.

  17. Interaction genotype by season and its influence on the identification of beans with high content of zinc and iron

    Directory of Open Access Journals (Sweden)

    Camila Andrade Silva

    2012-01-01

    Full Text Available The mineral contents in common bean seeds are influenced, in addition to genetic variation, by environmental crop conditions, especially by the soil type and chemical composition and by the genotype x environment interaction. This study was carried out to verify if the zinc and iron contents are affected by the crop growing period. Ten lines with high iron and zinc contents and ten with low contents were assessed in three seasons: "wet season" of 2009/2010 (sowing in November; "dry season" of 2010 (sowing in February and "winter season" of 2010 (sowing in July, in Lavras, Minas Gerais State, Brazil. The experimental design used was randomized blocks with three replications and plots consisting of two rows of two meters, with a spacing of 0.50 m. The seeds harvested were assessed in regard to iron and zinc mineral contents. The greatest contents were observed in the winter season and the smallest ones in the dry season, with sowing in February. It was observed that in the mean of the three harvests, the lines classified as having high iron and zinc content exhibited an iron quantity 11.0% and a zinc quantity 6.8% above those of low content. The lines by seasons interaction occurs. However, its interference in identification of the groups with high and low content of the two nutrients is not great.

  18. Simultaneous multi-parametric analysis of Leishmania and of its hosting mammal cells: A high content imaging-based method enabling sound drug discovery process.

    Science.gov (United States)

    Forestier, Claire-Lise; Späth, Gerald Frank; Prina, Eric; Dasari, Sreekanth

    2015-11-01

    Leishmaniasis is a vector-borne disease for which only limited therapeutic options are available. The disease is ranked among the six most important tropical infectious diseases and represents the second-largest parasitic killer in the world. The development of new therapies has been hampered by the lack of technologies and methodologies that can be integrated into the complex physiological environment of a cell or organism and adapted to suitable in vitro and in vivo Leishmania models. Recent advances in microscopy imaging offer the possibility to assess the efficacy of potential drug candidates against Leishmania within host cells. This technology allows the simultaneous visualization of relevant phenotypes in parasite and host cells and the quantification of a variety of cellular events. In this review, we present the powerful cellular imaging methodologies that have been developed for drug screening in a biologically relevant context, addressing both high-content and high-throughput needs. Furthermore, we discuss the potential of intra-vital microscopy imaging in the context of the anti-leishmanial drug discovery process.

  19. Systematic Characterization of Cell Cycle Phase-dependent Protein Dynamics and Pathway Activities by High-content Microscopy-assisted Cell Cycle Phenotyping

    Institute of Scientific and Technical Information of China (English)

    Christopher Bruhn; Torsten Kroll; Zhao-Qi Wang

    2014-01-01

    Cell cycle progression is coordinated with metabolism, signaling and other complex cel-lular functions. The investigation of cellular processes in a cell cycle stage-dependent manner is often the subject of modern molecular and cell biological research. Cell cycle synchronization and immunostaining of cell cycle markers facilitate such analysis, but are limited in use due to unphysiological experimental stress, cell type dependence and often low flexibility. Here, we describe high-content microscopy-assisted cell cycle phenotyping (hiMAC), which integrates high-resolution cell cycle profiling of asynchronous cell populations with immunofluorescence microscopy. hiMAC is compatible with cell types from any species and allows for statistically pow-erful, unbiased, simultaneous analysis of protein interactions, modifications and subcellular locali-zation at all cell cycle stages within a single sample. For illustration, we provide a hiMAC analysis pipeline tailored to study DNA damage response and genomic instability using a 3–4-day protocol, which can be adjusted to any other cell cycle stage-dependent analysis.

  20. Low Temperature Deposition of High-k/Metal Gate Stacks on High-Sn Content (Si)GeSn-Alloys.

    Science.gov (United States)

    Schulte-Braucks, C; von den Driesch, N; Glass, S; Tiedemann, A T; Breuer, U; Besmehn, A; Hartmann, J-M; Ikonic, Z; Zhao, Q T; Mantl, S; Buca, D

    2016-05-25

    (Si)GeSn is an emerging group IV alloy system offering new exciting properties, with great potential for low power electronics due to the fundamental direct band gap and prospects as high mobility material. In this Article, we present a systematic study of HfO2/TaN high-k/metal gate stacks on (Si)GeSn ternary alloys and low temperature processes for large scale integration of Sn based alloys. Our investigations indicate that SiGeSn ternaries show enhanced thermal stability compared to GeSn binaries, allowing the use of the existing Si technology. Despite the multielemental interface and large Sn content of up to 14 atom %, the HfO2/(Si)GeSn capacitors show small frequency dispersion and stretch-out. The formed TaN/HfO2/(Si)GeSn capacitors present a low leakage current of 2 × 10(-8) A/cm(2) at -1 V and a high breakdown field of ∼8 MV/cm. For large Sn content SiGeSn/GeSn direct band gap heterostructures, process temperatures below 350 °C are required for integration. We developed an atomic vapor deposition process for TaN metal gate on HfO2 high-k dielectric and validated it by resistivity as well as temperature and frequency dependent capacitance-voltage measurements of capacitors on SiGeSn and GeSn. The densities of interface traps are deduced to be in the low 10(12) cm(-2) eV(-1) range and do not depend on the Sn-concentration. The new processes developed here are compatible with (Si)GeSn integration in large scale applications.

  1. A multi-functional imaging approach to high-content protein interaction screening.

    Directory of Open Access Journals (Sweden)

    Daniel R Matthews

    Full Text Available Functional imaging can provide a level of quantification that is not possible in what might be termed traditional high-content screening. This is due to the fact that the current state-of-the-art high-content screening systems take the approach of scaling-up single cell assays, and are therefore based on essentially pictorial measures as assay indicators. Such phenotypic analyses have become extremely sophisticated, advancing screening enormously, but this approach can still be somewhat subjective. We describe the development, and validation, of a prototype high-content screening platform that combines steady-state fluorescence anisotropy imaging with fluorescence lifetime imaging (FLIM. This functional approach allows objective, quantitative screening of small molecule libraries in protein-protein interaction assays. We discuss the development of the instrumentation, the process by which information on fluorescence resonance energy transfer (FRET can be extracted from wide-field, acceptor fluorescence anisotropy imaging and cross-checking of this modality using lifetime imaging by time-correlated single-photon counting. Imaging of cells expressing protein constructs where eGFP and mRFP1 are linked with amino-acid chains of various lengths (7, 19 and 32 amino acids shows the two methodologies to be highly correlated. We validate our approach using a small-scale inhibitor screen of a Cdc42 FRET biosensor probe expressed in epidermoid cancer cells (A431 in a 96 microwell-plate format. We also show that acceptor fluorescence anisotropy can be used to measure variations in hetero-FRET in protein-protein interactions. We demonstrate this using a screen of inhibitors of internalization of the transmembrane receptor, CXCR4. These assays enable us to demonstrate all the capabilities of the instrument, image processing and analytical techniques that have been developed. Direct correlation between acceptor anisotropy and donor FLIM is observed for FRET

  2. Highly Parallel Computing Architectures by using Arrays of Quantum-dot Cellular Automata (QCA): Opportunities, Challenges, and Recent Results

    Science.gov (United States)

    Fijany, Amir; Toomarian, Benny N.

    2000-01-01

    There has been significant improvement in the performance of VLSI devices, in terms of size, power consumption, and speed, in recent years and this trend may also continue for some near future. However, it is a well known fact that there are major obstacles, i.e., physical limitation of feature size reduction and ever increasing cost of foundry, that would prevent the long term continuation of this trend. This has motivated the exploration of some fundamentally new technologies that are not dependent on the conventional feature size approach. Such technologies are expected to enable scaling to continue to the ultimate level, i.e., molecular and atomistic size. Quantum computing, quantum dot-based computing, DNA based computing, biologically inspired computing, etc., are examples of such new technologies. In particular, quantum-dots based computing by using Quantum-dot Cellular Automata (QCA) has recently been intensely investigated as a promising new technology capable of offering significant improvement over conventional VLSI in terms of reduction of feature size (and hence increase in integration level), reduction of power consumption, and increase of switching speed. Quantum dot-based computing and memory in general and QCA specifically, are intriguing to NASA due to their high packing density (10(exp 11) - 10(exp 12) per square cm ) and low power consumption (no transfer of current) and potentially higher radiation tolerant. Under Revolutionary Computing Technology (RTC) Program at the NASA/JPL Center for Integrated Space Microelectronics (CISM), we have been investigating the potential applications of QCA for the space program. To this end, exploiting the intrinsic features of QCA, we have designed novel QCA-based circuits for co-planner (i.e., single layer) and compact implementation of a class of data permutation matrices, a class of interconnection networks, and a bit-serial processor. Building upon these circuits, we have developed novel algorithms and QCA

  3. Highly Parallel Computing Architectures by using Arrays of Quantum-dot Cellular Automata (QCA): Opportunities, Challenges, and Recent Results

    Science.gov (United States)

    Fijany, Amir; Toomarian, Benny N.

    2000-01-01

    There has been significant improvement in the performance of VLSI devices, in terms of size, power consumption, and speed, in recent years and this trend may also continue for some near future. However, it is a well known fact that there are major obstacles, i.e., physical limitation of feature size reduction and ever increasing cost of foundry, that would prevent the long term continuation of this trend. This has motivated the exploration of some fundamentally new technologies that are not dependent on the conventional feature size approach. Such technologies are expected to enable scaling to continue to the ultimate level, i.e., molecular and atomistic size. Quantum computing, quantum dot-based computing, DNA based computing, biologically inspired computing, etc., are examples of such new technologies. In particular, quantum-dots based computing by using Quantum-dot Cellular Automata (QCA) has recently been intensely investigated as a promising new technology capable of offering significant improvement over conventional VLSI in terms of reduction of feature size (and hence increase in integration level), reduction of power consumption, and increase of switching speed. Quantum dot-based computing and memory in general and QCA specifically, are intriguing to NASA due to their high packing density (10(exp 11) - 10(exp 12) per square cm ) and low power consumption (no transfer of current) and potentially higher radiation tolerant. Under Revolutionary Computing Technology (RTC) Program at the NASA/JPL Center for Integrated Space Microelectronics (CISM), we have been investigating the potential applications of QCA for the space program. To this end, exploiting the intrinsic features of QCA, we have designed novel QCA-based circuits for co-planner (i.e., single layer) and compact implementation of a class of data permutation matrices, a class of interconnection networks, and a bit-serial processor. Building upon these circuits, we have developed novel algorithms and QCA

  4. Rapid RNA-ligand interaction analysis through high-information content conformational and stability landscapes

    Energy Technology Data Exchange (ETDEWEB)

    Baird, Nathan J. [National Inst. of Health (NIH), Bethesda, MD (United States); Inglese, James [National Inst. of Health (NIH), Bethesda, MD (United States); Ferré-D’Amaré, Adrian R. [National Inst. of Health (NIH), Bethesda, MD (United States)

    2015-12-07

    The structure and biological properties of RNAs are a function of changing cellular conditions, but comprehensive, simultaneous investigation of the effect of multiple interacting environmental variables is not easily achieved. We have developed an efficient, high-throughput method to characterize RNA structure and thermodynamic stability as a function of multiplexed solution conditions using Förster resonance energy transfer (FRET). In a single FRET experiment using conventional quantitative PCR instrumentation, 19,400 conditions of MgCl2, ligand and temperature are analysed to generate detailed empirical conformational and stability landscapes of the cyclic diguanylate (c-di-GMP) riboswitch. This method allows rapid comparison of RNA structure modulation by cognate and non-cognate ligands. Landscape analysis reveals that kanamycin B stabilizes a non-native, idiosyncratic conformation of the riboswitch that inhibits c-di-GMP binding. Our research demonstrates that allosteric control of folding, rather than direct competition with cognate effectors, is a viable approach for pharmacologically targeting riboswitches and other structured RNA molecules.

  5. High-Content Movement Analysis as a Diagnostic Tool in C. elegans

    Science.gov (United States)

    Winter, Peter; Lancichinetti, Andrea; Krevitt, Leah; Amaral, Luis; Morimoto, Rick

    2013-03-01

    Many neurodegenerative diseases manifest themselves through a loss of motor control and give us information about the underlying disease. This loss of coordination is observed in humans and in the model organisms used to study neurodegeneration. In Caenorhabditis elegans, there is an extensive genetic library of strains that lack functional neuronal signaling pathways and expressing proteins associated with neurodegenerative diseases. While most of these strains have decrease motility or cause paralysis, relatively few have been screened to look for more subtle changes in motor control such as stiffness, twitching, or other changes in behavior. we use high-resolution position and posture data to automatically analyze the movement of worms from different genetic backgrounds and characterize 14 movement characteristics. By creating a quantitative mapping between the movement characterization and an online database of gene annotation, gene expression, and anatomy, we aim to predict a likely set of cellular and molecular disruptions. This work provides a proof of concept for the use of detailed movement analysis to uncover novel disruptions in certain motor control processes. Knowledge of the molecular origin of these disruptions provided by our understanding of C. elegans genetics and physiology could lead to new diagnostic and therapeutic targets for neurodegenerative disease.

  6. Rapid RNA-ligand interaction analysis through high-information content conformational and stability landscapes

    Science.gov (United States)

    Baird, Nathan J.; Inglese, James; Ferré-D'Amaré, Adrian R.

    2015-12-01

    The structure and biological properties of RNAs are a function of changing cellular conditions, but comprehensive, simultaneous investigation of the effect of multiple interacting environmental variables is not easily achieved. We have developed an efficient, high-throughput method to characterize RNA structure and thermodynamic stability as a function of multiplexed solution conditions using Förster resonance energy transfer (FRET). In a single FRET experiment using conventional quantitative PCR instrumentation, 19,400 conditions of MgCl2, ligand and temperature are analysed to generate detailed empirical conformational and stability landscapes of the cyclic diguanylate (c-di-GMP) riboswitch. The method allows rapid comparison of RNA structure modulation by cognate and non-cognate ligands. Landscape analysis reveals that kanamycin B stabilizes a non-native, idiosyncratic conformation of the riboswitch that inhibits c-di-GMP binding. This demonstrates that allosteric control of folding, rather than direct competition with cognate effectors, is a viable approach for pharmacologically targeting riboswitches and other structured RNA molecules.

  7. High water contents in basaltic melt inclusions from Arenal volcano, Costa Rica

    Science.gov (United States)

    Wade, J. A.; Plank, T.; Hauri, E. H.; Melson, W. G.; Soto, G. J.

    2004-12-01

    Despite the importance of water to arc magma genesis, fractionation and eruption, few quantitative constraints exist on the water content of Arenal magmas. Early estimates, by electron microprobe sum deficit, suggested up to 4 wt% H2O in olivine-hosted basaltic andesite melt inclusions (MI) from pre-historic ET-6 tephra (Melson, 1982), and up to 7 wt% H2O in plagioclase and orthopyroxene-hosted dacitic MI from 1968 lapilli (Anderson, 1979). These high water contents are consistent with abundant hornblende phenocrysts in Arenal volcanics, but inconsistent with geochemical tracers such as 10Be and Ba/La that suggest a low flux of recycled material (and presumably water) from the subduction zone. In order to test these ideas, and provide the first direct measurements of water in mafic Arenal magmas, we have studied olivine-hosted MI from the prehistoric (900 yBP; Soto et al., 1998) ET3 tephra layer. MI range from andesitic (> 58% SiO2) to basaltic compositions ( 4 wt%) found here for Arenal basaltic MI support the semi-quantitative data from earlier studies, but are somewhat unexpected given predictions from slab tracers. Arenal water contents (4%) approach those of the 1995 eruption of Cerro Negro in Nicaragua (4-5 wt% in basaltic MI; Roggensack et al., 1997), despite the fact that the latter has Ba/La of > 100, while Arenal has Ba/La Boletin de Volcanologia; Roggensack et al. (1997) Science; Soto et al. (1998) OSIVAM; Williams-Jones et al. (2001) Journal of Volc. and Geoth. Res.

  8. Noble strain of Sparassis latifolia produces high content of β-glucan

    Institute of Scientific and Technical Information of China (English)

    Dong; Ju; Lee; Min; Cheol; Jang; A; Ra; Jo; Hyun; Jun; Choi; Kwang-Sang; Kim; Youn-Tae; Chi

    2015-01-01

    Objective: To classify four new Sparassis strains(CLM1, CKM1, CKM2, and KJM1) using the internal transcribed spacer sequence and to elucidate their β-glucan content and mycelial growth.Methods: Two different microbiological media were used to determine growth rate. The β-glucan contents were analyzed using the Megazyme Mushroom and Yeast Beta-Glucan kit. To determine the genetic relationships, phylogenetic trees were constructed using ClustalX. Multiple sequence alignments were printed and shaded with the BOXSHADE 3.21 program. Results: In this study, four new Sparassis strains were isolated from the southern region of the Korea Peninsula. They were all classified into the Sparassis latifolia clade as a monophyletic group based on the internal transcribed spacer sequence. Mycelial growth rate of the CLM1 strain was highest in potato dextrose agar and potato dextrose agar larch. The β-glucan content of the CLM1 strain was highest at 29.5%(w/w). A high degree of sequence divergence was detected in the RNA polymerase second largest subunit II gene(RPB2) within Sparassis spp. tested. The putative amino acid sequences of the RPB2 had a distinct sequence. The nucleotide sequences of the RPB2’s intron were also divergent among Sparassis spp., even though their nucleotide length was well conserved within Sparassis latifolia. Conclusions: These results indicate that the nucleotide sequences and the amino acid sequences of RPB2 can be used to identify individual Sparassis sp. The Sparassis strain CLM1 may be best for developing a remedy to prevent or treat cancer and other chronic diseases.

  9. Fat content increases the lethality of ultra-high-pressure homogenization on Listeria monocytogenes in milk.

    Science.gov (United States)

    Roig-Sagués, A X; Velázquez, R M; Montealegre-Agramont, P; López-Pedemonte, T J; Briñez-Zambrano, W J; Guamis-López, B; Hernandez-Herrero, M M

    2009-11-01

    Listeria monocytogenes CCUG 15526 was inoculated at a concentration of approximately 7.0 log(10) cfu/mL in milk samples with 0.3, 3.6, 10, and 15% fat contents. Milk samples with 0.3 and 3.6% fat content were also inoculated with a lower load of approximately 3.0 log(10) cfu/mL. Inoculated milk samples were subjected to a single cycle of ultra-high-pressure homogenization (UHPH) treatment at 200, 300, and 400 MPa. Microbiological analyses were performed 2 h after the UHPH treatments and after 5, 8, and 15 d of storage at 4 degrees C. Maximum lethality values were observed in samples treated at 400 MPa with 15 and 10% fat (7.95 and 7.46 log(10) cfu/mL), respectively. However, in skimmed and 3.6% fat milk samples, complete inactivation was not achieved and, during the subsequent 15 d of storage at 4 degrees C, L. monocytogenes was able to recover and replicate until achieving initial counts. In milk samples with 10 and 15% fat, L. monocytogenes recovered to the level of initial counts only in the milk samples treated at 200 MPa but not in the milk samples treated at 300 and 400 MPa. When the load of L. monocytogenes was approximately 3.0 log(10) cfu/mL in milk samples with 0.3 and 3.6% fat, complete inactivation was not achieved and L. monocytogenes was able to recover and grow during the subsequent cold storage. Fat content increased the maximum temperature reached during UHPH treatment; this could have contributed to the lethal effect achieved, but the amount of fat of the milk had a stronger effect than the temperature on obtaining a higher death rate of L. monocytogenes.

  10. High-resolution prediction of soil available water content within the crop root zone

    Science.gov (United States)

    Haghverdi, Amir; Leib, Brian G.; Washington-Allen, Robert A.; Ayers, Paul D.; Buschermohle, Michael J.

    2015-11-01

    A detailed understanding of soil hydraulic properties, particularly soil available water content (AWC) within the effective root zone, is needed to optimally schedule irrigation in fields with substantial spatial heterogeneity. However, it is difficult and time consuming to directly measure soil hydraulic properties. Therefore, easily collected and measured soil properties, such as soil texture and/or bulk density, that are well correlated with hydraulic properties are used as proxies to develop pedotransfer functions (PTF). In this study, multiple modeling scenarios were developed and evaluated to indirectly predict high resolution AWC maps within the effective root zone. The modeling techniques included kriging, co-kriging, regression kriging, artificial neural networks (NN) and geographically weighted regression (GWR). The efficiency of soil apparent electrical conductivity (ECa) as proximal data in the modeling process was assessed. There was a good agreement (root mean square error (RMSE) = 0.052 cm3 cm-3 and r = 0.88) between observed and point prediction of water contents using pseudo continuous PTFs. We found that both GWR (mean RMSE = 0.062 cm3 cm-3) and regression kriging (mean RMSE = 0.063 cm3 cm-3) produced the best water content maps with these accuracies improved up to 19% when ECa was used as an ancillary soil attribute in the interpolation process. The maps indicated fourfold differences in AWC between coarse- and fine-textured soils across the study site. This provided a template for future investigations for evaluating the efficiency of variable rate irrigation management scenarios in accounting for the spatial heterogeneity of soil hydraulic attributes.

  11. Cationic Copolymerization of 3,3-Bis(hydroxymethyl)oxetane and Glycidol: Biocompatible Hyperbranched Polyether Polyols with High Content of Primary Hydroxyl Groups.

    Science.gov (United States)

    Christ, Eva-Maria; Hobernik, Dominika; Bros, Matthias; Wagner, Manfred; Frey, Holger

    2015-10-12

    The cationic ring-opening copolymerization of 3,3-bis(hydroxymethyl)oxetane (BHMO) with glycidol using different comonomer ratios (BHMO content from 25 to 90%) and BF3OEt2 as an initiator has been studied. Apparent molecular weights of the resulting hyperbranched polyether copolymers ranged from 1400 to 3300 g mol(-1) (PDI: 1.21-1.48; method: SEC, linear PEG standards). Incorporation of both comonomers is evidenced by MALDI-TOF mass spectroscopy. All hyperbranched polyether polyols with high content of primary hydroxyl groups portray good solubility in water, which correlates with an increasing content of glycerol units. Detailed NMR characterization was employed to elucidate the copolymer microstructures. Kinetic studies via FTIR demonstrated a weak gradient-type character of the copolymers. MTT assays of the copolymers (up to 100 μg mL(-1)) on HEK and fibroblast cell lines (3T3, L929, WEHI) as well as viability tests on the fibroblast cells were carried out to assess the biocompatibility of the materials, confirming excellent biocompatibility. Transfection efficiency characterization by flow cytometry and confocal laser microscopy demonstrated cellular uptake of the copolymers. Antiadhesive properties of the materials on surfaces were assessed by adhesion assays with fibroblast cells.

  12. Vanillin inhibits translation and induces messenger ribonucleoprotein (mRNP) granule formation in saccharomyces cerevisiae: application and validation of high-content, image-based profiling.

    Science.gov (United States)

    Iwaki, Aya; Ohnuki, Shinsuke; Suga, Yohei; Izawa, Shingo; Ohya, Yoshikazu

    2013-01-01

    Vanillin, generated by acid hydrolysis of lignocellulose, acts as a potent inhibitor of the growth of the yeast Saccharomyces cerevisiae. Here, we investigated the cellular processes affected by vanillin using high-content, image-based profiling. Among 4,718 non-essential yeast deletion mutants, the morphology of those defective in the large ribosomal subunit showed significant similarity to that of vanillin-treated cells. The defects in these mutants were clustered in three domains of the ribosome: the mRNA tunnel entrance, exit and backbone required for small subunit attachment. To confirm that vanillin inhibited ribosomal function, we assessed polysome and messenger ribonucleoprotein granule formation after treatment with vanillin. Analysis of polysome profiles showed disassembly of the polysomes in the presence of vanillin. Processing bodies and stress granules, which are composed of non-translating mRNAs and various proteins, were formed after treatment with vanillin. These results suggest that vanillin represses translation in yeast cells.

  13. Auxiliary units for refining of high nitrogen content oils: Premium II refinery case

    Energy Technology Data Exchange (ETDEWEB)

    Nicolato, Paolo Contim; Pinotti, Rafael [Petroleo Brasileiro S.A. (PETROBRAS), Rio de Janeiro, RJ (Brazil)

    2012-07-01

    PETROBRAS is constantly investing on its refining park in order to increase the production of clean and stable fuels and to be capable to process heavier oils with high contaminants content. Sulfur and nitrogen are the main heteroatoms present in petroleum. They are responsible for some undesirable fuels properties like corrosivity and instability, and also emit pollutants when burnt. Hydrotreating and hydrocracking processes are designed to remove these contaminants and adjust other fuel properties, generating, as byproduct, sour gases and sour water streams rich in H{sub 2}S and NH{sub 3}, which are usually sent to Sour Water Treatment Units and Sulfur Recovery Units. The regeneration of the amine used for the light streams treatment, as fuel gas and LPG, also generates sour gas streams that must be also sent to Sulfur Recovery Units. As the ammonia content in the sour streams increases, some design parameters must be adjusted to avoid increasing the Refinery emissions. Sulfur Recovery Units must provide proper NH3 destruction. Sour Water Treatment must have a proper segregation between H{sub 2}S and ammonia streams, whenever desirable. Amine Regeneration Systems must have an efficient procedure to avoid the ammonia concentration in the amine solution. This paper presents some solutions usually applied to the Petroleum Industry and analyses some aspects related to Premium II Refinery Project and how its design will help the Brazilian refining park to meet future environmental regulation and market demands. (author)

  14. Live Broadcasting of High Definition Audiovisual Content Using HDTV over Broadband IP Networks

    Directory of Open Access Journals (Sweden)

    C. E. Vegiris

    2008-01-01

    Full Text Available The current paper focuses on validating an implementation of a state-of-the art audiovisual (AV technologies setup for live broadcasting of cultural shows, via broadband Internet. The main objective of the work was to study, configure, and setup dedicated audio-video equipment for the processes of capturing, processing, and transmission of extended resolution and high fidelity AV content in order to increase realism and achieve maximum audience sensation. Internet2 and GEANT broadband telecommunication networks were selected as the most applicable technology to deliver such traffic workloads. Validation procedures were conducted in combination with metric-based quality of service (QoS and quality of experience (QoE evaluation experiments for the quantification and the perceptual interpretation of the quality achieved during content reproduction. The implemented system was successfully applied in real-world applications, such as the transmission of cultural events from Thessaloniki Concert Hall throughout Greece as well as the reproduction of Philadelphia Orchestra performances (USA via Internet2 and GEANT backbones.

  15. Effect of ammonium concentration on alcoholic fermentation kinetics by wine yeasts for high sugar content.

    Science.gov (United States)

    Taillandier, Patricia; Ramon Portugal, Felipe; Fuster, André; Strehaiano, Pierre

    2007-02-01

    Kinetics of alcoholic fermentation by Saccharomyces cerevisiae wine strains in a synthetic medium with high sugar content were established for different nitrogen initial content and are presented for four strains. The composition of the medium was close to grape must except that the nitrogen source consisted mainly in ammonium and was varied from 120 to 290 mg N/l assimilable nitrogen. The overall nitrogen consumed was also estimated in order to determine nitrogen requirement variability. The effect of assimilable nitrogen was in general greater on sugar consumption rates than on growth and three kinds of effect on sugar consumption rates were observed: (i) existence of an optimal initial nitrogen level for a maximal sugar consumption rate (inhibition if excess), (ii) no effect of nitrogen beyond the intermediary level (saturation), (iii) sugar consumption rate proportional to the initial nitrogen level (activation). In all cases, the amount of consumed nitrogen increased with its initial concentration and so did the fructophilic capacity of the strains. The optimal requirement varied from 0.62 to 0.91 mg N/g of sugars according to different strains. There was no general correlation between the sugar assimilation rates and the nitrogen requirement.

  16. Fuel-N Evolution during the Pyrolysis of Industrial Biomass Wastes with High Nitrogen Content

    Directory of Open Access Journals (Sweden)

    Kunio Yoshikawa

    2012-12-01

    Full Text Available In this study, sewage sludge and mycelial waste from antibiotic production were pyrolyzed in a batch scale fixed-bed reactor as examples of two kinds of typical industrial biomass wastes with high nitrogen content. A series of experiments were conducted on the rapid pyrolysis and the slow pyrolysis of these wastes in the temperature range from 500–800 °C to investigate the Fuel-N transformation behavior among pyrolysis products. The results showed that Fuel-N conversion to Char-N intimately depended on the pyrolysis temperature and the yield of Char-N reduced with the increase of the pyrolysis temperature. Under the same pyrolysis conditions, Tar-N production mainly depended on complex properties of the different biomasses, including volatile matter, nitrogen content and biomass functional groups. HCN was the predominant NOx precursor in the rapid pyrolysis of biomass, whereas in the slow pyrolysis of mycelial waste, more NH3 was produced than HCN due to the additional NH3 formation through the hydrogenation reaction of Char-N, HCN and H radicals. At the same time, some part of the char was analyzed by Fourier Transform infrared spectroscopy (FTIR to get more information on the nitrogen functionality changes and the tar was also characterized by Gas Chromatography and Mass Spectrometry (GCMS to identify typical nitrogenous tar compounds. Finally, the whole nitrogen distribution in products was discussed.

  17. Processing and Characterization of High Density Polyethylene/Ethylene Vinyl Acetate Blends with Different VA Contents

    Directory of Open Access Journals (Sweden)

    Othman Y. Alothman

    2012-01-01

    Full Text Available Different series of high density Polyethylene/Ethylene Vinyl Acetate (HDPE/EVA blends were prepared via melt blending in a corotating intermeshing twin screw extruder. The effects of VA percentage and EVA loading ratio on the thermal, rheological viscoelastic, mechanical, and fracture toughness of the blends were analyzed. The results showed that the addition of EVA to HDPE reduces the thermal, elastic, and viscoelastic properties of the blends. The microscopic examination of the fracture surface confirmed the ductile fracture of HDPE/EVA blends for all blend ratios and VA percentages. Increasing the EVA ratio and VA content caused a significant reduction in the blend crystallinity but had no significant effect on melting temperature. The complex viscosity increased with increasing the percentage of EVA due to the restriction of molecular mobility and reduction of free volume, induced by the addition of EVA. The storage modulus decreased with increasing the EVA ratio and temperature, while it increased with increasing the frequency. Young’s modulus, yield strength, and fracture strain decreased with increasing the EVA ratio. Similarly, the fracture toughness decreased proportional to the EVA percentage. Finally the results indicated that the VA content has significant effects on the mechanical, thermal, and dynamic properties of HDPE/EVA blends.

  18. Protein adsorption of dialdehyde cellulose-crosslinked chitosan with high amino group contents.

    Science.gov (United States)

    Kim, Ung-Jin; Lee, Yeong Ro; Kang, Tong Ho; Choi, Joon Weon; Kimura, Satoshi; Wada, Masahisa

    2017-05-01

    Crosslinked chitosan was prepared by Schiff base formation between the aldehyde groups of dialdehyde cellulose (DAC) and the amino groups of chitosan and a subsequent reduction. DAC was obtained through periodate oxidation of cellulose and solubilization in hot water at 100°C for 1h. Three grades of DAC-crosslinked chitosan were prepared by adding various amounts DAC. The degrees of crosslinking as determined by amino group content were 3.8, 8.3, and 12.1%, respectively. DAC-crosslinked chitosan showed higher stability in the pH 2-9 range and no cytotoxicity was identified over the course of a 21-day long-term stability test. Also, DAC-crosslinked chitosan showed remarkably high bovine serum albumin (BSA) adsorption capacity at pH 5.5 as a result of the increased amino group content, due to the reaction between DAC and chitosan molecular chains occurring at multiple points even though DAC-crosslinked chitosan showed a lower degree of crosslinking.

  19. Local immunosuppression induced by high viral load of human papillomavirus: characterization of cellular phenotypes producing interleukin-10 in cervical neoplastic lesions.

    Science.gov (United States)

    Prata, Thiago Theodoro Martins; Bonin, Camila Mareti; Ferreira, Alda Maria Teixeira; Padovani, Cacilda Tezelli Junqueira; Fernandes, Carlos Eurico dos Santos; Machado, Ana Paula; Tozetti, Inês Aparecida

    2015-09-01

    A specific immune response to human papillomavirus (HPV) in the cervical microenvironment plays a key role in eradicating infection and eliminating mutated cells. However, high-risk HPVs modulate immune cells to create an immunosuppressive microenvironment, and induce these immune cells to produce interleukin 10 (IL-10). This production of IL-10, in conjunction with HPV infection, contributes to the appearance of cervical neoplastic lesions. We sought to characterize the IL-10-producing cellular phenotype, and investigate the influence of host and HPV factors upon the induction of an immunosuppressive microenvironment. Immunohistochemical analysis demonstrated an increase in IL-10 production by keratinocytes, macrophages and Langerhans cells in high-grade cervical lesions and cervical cancer. This increase was more pronounced in patients older than 30 years, and was also correlated with high viral load, and infection with a single HPV type, particularly high-risk HPVs. Our results indicate the existence of a highly immunosuppressive microenvironment composed of different IL-10-producing cellular phenotypes in cervical cancer samples, and samples classified as high-grade cervical lesions (cervical intraepithelial neoplasia stages II and III). The immunosuppressive microenvironment that developed for these different cellular phenotypes favours viral persistence and neoplastic progression.

  20. Covalent Triazine-Based Frameworks with Ultramicropores and High Nitrogen Contents for Highly Selective CO2 Capture.

    Science.gov (United States)

    Wang, Keke; Huang, Hongliang; Liu, Dahuan; Wang, Chang; Li, Jinping; Zhong, Chongli

    2016-05-03

    Porous organic frameworks (POFs) are a class of porous materials composed of organic precursors linked by covalent bonds. The objective of this work is to develop POFs with both ultramicropores and high nitrogen contents for CO2 capture. Specifically, two covalent triazine-based frameworks (CTFs) with ultramicropores (pores of width capture CO2 due to ultramicroporous nature. Especially, CTF-FUM-350 has the highest nitrogen content (27.64%) and thus the highest CO2 adsorption capacity (57.2 cc/g at 298 K) and selectivities for CO2 over N2 and CH4 (102.4 and 20.5 at 298 K, respectively) among all CTF-FUM and CTF-DCN. More impressively, as far as we know, the CO2/CH4 selectivity is larger than that of all reported CTFs and ranks in top 10 among all reported POFs. Dynamic breakthrough curves indicate that both CTFs could indeed separate gas mixtures of CO2/N2 and CO2/CH4 completely.

  1. Analysis of Information Content in High-Spectral Resolution Sounders using Subset Selection Analysis

    Science.gov (United States)

    Velez-Reyes, Miguel; Joiner, Joanna

    1998-01-01

    In this paper, we summarize the results of the sensitivity analysis and data reduction carried out to determine the information content of AIRS and IASI channels. The analysis and data reduction was based on the use of subset selection techniques developed in the linear algebra and statistical community to study linear dependencies in high dimensional data sets. We applied the subset selection method to study dependency among channels by studying the dependency among their weighting functions. Also, we applied the technique to study the information provided by the different levels in which the atmosphere is discretized for retrievals and analysis. Results from the method correlate well with intuition in many respects and point out to possible modifications for band selection in sensor design and number and location of levels in the analysis process.

  2. THE CRYSTALLIZATION OF POTASSIUM GERMANATE GLASS WITH HIGH CONTENT OF NIOBIUM OXIDE

    Directory of Open Access Journals (Sweden)

    SRĐAN D.MATIJAŠEVIĆ

    2012-03-01

    Full Text Available Potassium germanate glass with molar ratio [GeO2]/[K2O] = 1.2 and Nb2O5 content of 34 mol% have been synthesized by a melt-quenching method. The crystallization behavior under non-isothermal and isothermal crystallization conditions was investigated. The results showed that this glass exposed complex primary crystallization. In the temperature range 800 °C. K10Nb22Ge4O68 and metastable KNbO3 and K4Nb6O17 were formed as secondary phases. The crystallization commenced at T > 640 °C with high homogeneous nucleation rate and spherulite crystal growth morphology. DTA curves recorded for powder samples particle size 0.1 mm showed two exothermic peaks and two endothermic peaks within temperature range of T = 640-1020 °C. The analysis of the dominant crystallization mechanism of powder glass sample and kinetics of crystallization is presented.

  3. Bench-scale synthesis of zeolite A from subbituminous coal ashes with high crystalline silica content

    Energy Technology Data Exchange (ETDEWEB)

    Chareonpanich, M.; Jullaphan, O.; Tang, C. [Kasetsart University, Bangkok (Thailand). Dept. of Chemical Engineering

    2011-01-15

    In this present work, fly ash and bottom ash with high crystalline silica content were obtained from the coal-fired boilers within the paper industries in Thailand. These coal ashes were used as the basic raw materials for synthetic zeolite production. The crystal type and crystallinity, specific surface area and pore size, and textural properties of zeolite products were characterized by using X-ray diffraction spectroscopy (XRD), N{sub 2} sorption analysis, and Scanning Electron Microscopy (SEM), respectively. It was found that sodalite octahydrate was selectively formed via the direct conventional (one-step) synthesis, whereas through a two-step, sodium silicate preparation and consecutive zeolite A synthesis process, 94 and 72 wt.% zeolite A products could be produced from the fly ash and bottom ash, respectively. The cation-exchange capacity (CEC) of fly ash and bottom ash-derived zeolite A products were closely similar to that of the commercial grade zeolite A.

  4. Process modules for GeSn nanoelectronics with high Sn-contents

    Science.gov (United States)

    Schulte-Braucks, C.; Glass, S.; Hofmann, E.; Stange, D.; von den Driesch, N.; Hartmann, J. M.; Ikonic, Z.; Zhao, Q. T.; Buca, D.; Mantl, S.

    2017-02-01

    This paper systematically studies GeSn n-FETs, from individual process modules to a complete device. High-k gate stacks and NiGeSn metallic contacts for source and drain are characterized in independent experiments. To study both direct and indirect bandgap semiconductors, a range of 0-14.5 at.% Sn-content GeSn alloys are investigated. Special emphasis is placed on capacitance-voltage (C-V) characteristics and Schottky-barrier optimization. GeSn n-FET devices are presented including temperature dependent I-V characteristics. Finally, as an important step towards implementing GeSn in tunnel-FETs, negative differential resistance in Ge0.87Sn0.13 tunnel-diodes is demonstrated at cryogenic temperatures. The present work provides a base for further optimization of GeSn FETs and novel tunnel FET devices.

  5. High solids content, soap-free, film-forming latexes stabilized by laponite clay platelets.

    Science.gov (United States)

    Bourgeat-Lami, Elodie; Guimarães, Thiago Rodrigues; Pereira, Ana Maria Cenacchi; Alves, Gizelda Maria; Moreira, José Carlos; Putaux, Jean-Luc; Dos Santos, Amilton Martins

    2010-11-01

    High solids content film-forming poly[styrene-co-(n-butyl acrylate)] [poly(Sty-co-BuA)] latexes armored with Laponite clay platelets have been synthesized by soap-free emulsion copolymerization of styrene and n-butyl acrylate. The polymerizations were performed in batch in the presence of Laponite and a methyl ether acrylate-terminated poly(ethylene glycol) macromonomer in order to promote polymer/clay association. The overall polymerization kinetics showed a pronounced effect of clay on nucleation and stabilization of the latex particles. Cryo-transmission electron microscopy observation confirmed the armored morphology and indicated that the majority of Laponite platelets were located at the particle surface. The resulting nanostructured films displayed enhanced mechanical properties.

  6. Colloidal processing of Fe-based metal ceramic composites with high content of ceramic reinforcement

    Energy Technology Data Exchange (ETDEWEB)

    Escribano, J. A.; Ferrari, B.; Alvaredo, P.; Gordo, E.; Sanchez-Herencia, A. J.

    2013-07-01

    Major difficulties of processing metal-matrix composites by means of conventional powder metallurgy techniques are the lack of dispersion of the phases within the final microstructure. In this work, processing through colloidal techniques of the Fe-based metal-matrix composites, with a high content of a ceramic reinforcement (Ti(C,N) ), is presented for the first time in the literature. The colloidal approach allows a higher control of the powders packing and a better homogenization of phases since powders are mixed in a liquid medium. The chemical stability of Fe in aqueous medium determines the dispersion conditions of the mixture. The Fe slurries were formulated by optimising their zeta potential and their rheology, in order to shape bulk pieces by slip-casting. Preliminary results demonstrate the viability of this procedure, also opening new paths to the microstructural design of fully sintered Fe-based hard metal, with 50 vol. % of Ti(C,N) in its composition. (Author)

  7. tranSMART: An Open Source Knowledge Management and High Content Data Analytics Platform.

    Science.gov (United States)

    Scheufele, Elisabeth; Aronzon, Dina; Coopersmith, Robert; McDuffie, Michael T; Kapoor, Manish; Uhrich, Christopher A; Avitabile, Jean E; Liu, Jinlei; Housman, Dan; Palchuk, Matvey B

    2014-01-01

    The tranSMART knowledge management and high-content analysis platform is a flexible software framework featuring novel research capabilities. It enables analysis of integrated data for the purposes of hypothesis generation, hypothesis validation, and cohort discovery in translational research. tranSMART bridges the prolific world of basic science and clinical practice data at the point of care by merging multiple types of data from disparate sources into a common environment. The application supports data harmonization and integration with analytical pipelines. The application code was released into the open source community in January 2012, with 32 instances in operation. tranSMART's extensible data model and corresponding data integration processes, rapid data analysis features, and open source nature make it an indispensable tool in translational or clinical research.

  8. Organosilica hybrid nanomaterials with a high organic content: syntheses and applications of silsesquioxanes

    KAUST Repository

    Croissant, Jonas G.

    2016-11-07

    Organic-inorganic hybrid materials garner properties from their organic and inorganic matrices as well as synergistic features, and therefore have recently attracted much attention at the nanoscale. Non-porous organosilica hybrid nanomaterials with a high organic content such as silsesquioxanes (R-SiO, with R organic groups) and bridged silsesquioxanes (OSi-R-SiO) are especially attractive hybrids since they provide 20 to 80 weight percent of organic functional groups in addition to the known chemistry and stability of silica. In the organosilica family, silsesquioxanes (R-SiO) stand between silicas (SiO) and silicones (RSiO), and are variously called organosilicas, ormosil (organically-modified silica), polysilsesquioxanes and silica hybrids. Herein, we comprehensively review non-porous silsesquioxane and bridged silsesquioxane nanomaterials and their applications in nanomedicine, electro-optics, and catalysis.

  9. Subfamily logos: visualization of sequence deviations at alignment positions with high information content

    Directory of Open Access Journals (Sweden)

    Beitz Eric

    2006-06-01

    Full Text Available Abstract Background Recognition of relevant sequence deviations can be valuable for elucidating functional differences between protein subfamilies. Interesting residues at highly conserved positions can then be mutated and experimentally analyzed. However, identification of such sites is tedious because automated approaches are scarce. Results Subfamily logos visualize subfamily-specific sequence deviations. The display is similar to classical sequence logos but extends into the negative range. Positive, upright characters correspond to residues which are characteristic for the subfamily, negative, upside-down characters to residues typical for the remaining sequences. The symbol height is adjusted to the information content of the alignment position. Residues which are conserved throughout do not appear. Conclusion Subfamily logos provide an intuitive display of relevant sequence deviations. The method has proven to be valid using a set of 135 aligned aquaporin sequences in which established subfamily-specific positions were readily identified by the algorithm.

  10. MALDI-TOF MS for quality control of high protein content sport supplements.

    Science.gov (United States)

    De Ceglie, Cristina; Calvano, Cosima D; Zambonin, Carlo G

    2015-06-01

    High protein content sport nutritional supplements are found as powder products containing, as ingredients, amino acids and proteins with important nutritional values as milk, soy and egg proteins. An EU Food Supplements Directive (2002) requires that supplements should be safe, both in dosages and in purity. It is important, then, to develop rapid and sensitive methods to be employed for the quality control of these substances. In this work, we apply, for the first time, matrix-assisted laser desorption ionization-mass spectrometry as a fast, reproducible and sensitive method for the quality control of sport nutritional supplements based on proteins. To this aim, several commercial egg- and/or milk-based powder products have been processed by in gel or in solution digestion and analyzed in comparison to pure standard products. This strategy allowed to assess the reliability of the indications on proteins (as caseins, whey proteins and ovalbumin) declared in the label of several sport nutritional supplements.

  11. High Stellar FUV/NUV Ratio and Oxygen Contents in the Atmospheres of Potentially Habitable Planets

    CERN Document Server

    Tian, Feng; Linsky, Jeffrey L; Mauas, Pablo J D; Vieytes, Mariela C

    2013-01-01

    Recent observations of several planet-hosting M dwarfs show that most have FUV/NUV flux ratios 1000 times greater than that of the Sun. Here we show that the atmospheric oxygen contents (O2 and O3) of potentially habitable planets in this type of UV environment could be 2~3 orders of magnitude greater than those of their counterparts around Sun-like stars as a result of decreased photolysis of O3, H2O2, and HO2. Thus detectable levels of atmospheric oxygen, in combination with the existence of H2O and CO2, may not be the most promising biosignatures on planets around stars with high FUV/NUV ratios such as the observed M dwarfs.

  12. Understanding a High School Physics Teacher's Pedagogical Content Knowledge of Argumentation

    Science.gov (United States)

    Wang, Jianlan; Buck, Gayle A.

    2016-08-01

    Scientific argumentation is an important learning objective in science education. It is also an effective instructional approach to constructivist science learning. The implementation of argumentation in school settings requires science teachers, who are pivotal agents of transforming classroom practices, to develop sophisticated knowledge of argumentation. However, there is a lack of understanding about science teachers' knowledge of argumentation, especially the dialogic meaning of argumentation. In this case study, we closely examine a high school physics teacher's argumentation-related pedagogic content knowledge (PCK) in the context of dialogic argumentation. We synthesize the teacher's performed PCK from his argumentation practices and narrated PCK from his reflection on the argumentation practices, from which we summarize his PCK of argumentation from the perspectives of orientation, instructional strategies, students, curriculum, and assessment. Finally, we describe the teacher's perception and adaption of argumentation in his class. We also identity the barriers to argumentation implementation in this particular case and suggest solutions to overcome these barriers.

  13. Modeling of Flow Stress of High Titanium Content 6061 Aluminum Alloy Under Hot Compression

    Science.gov (United States)

    Chen, Wei; Guan, Yingping; Wang, Zhenhua

    2016-09-01

    Hot compression tests were performed on high titanium content 6061 aluminum alloy (AA 6061-Ti) using a Gleeble-3500 thermomechanical testing system at temperatures from 350 to 510 °C with a constant strain rate in the range of 0.001-10 s-1. Three types of flow stress models were established from the experimental stress-strain curves, the correlation coefficient ( R), mean absolute relative error ( MARE), and root mean square deviation ( RMSD) between the predicted data and the experimental data were also calculated. The results show that the Fields-Backofen model, which includes a softening factor, was the simplest mathematical expression with a level of precision appropriate for the numerical simulations. However, the Arrhenius and artificial neural network (ANN) models were also consistent with the experimental results but they are more limited in their application in terms of their accuracy and the mathematical expression of the models.

  14. Doping of GaN{sub 1-x}As{sub x} with high As content

    Energy Technology Data Exchange (ETDEWEB)

    Levander, A.X.; Novikov, S.V.; Liliental-Weber, Z.; dos Reis, R.; Dubon, O.D.; Wu, J.; Foxon, C.T.; Yu, K.M.; Walukiewicz, W.

    2011-09-22

    Recent work has shown that GaN{sub 1-x}As{sub x} can be grown across the entire composition range by low temperature molecular beam epitaxy with intermediate compositions being amorphous, but control of the electrical properties through doping is critical for functionalizing this material. Here we report the bipolar doping of GaN{sub 1-x}As{sub x} with high As content to conductivities above 4 S/cm at room temperature using Mg or Te. The carrier type was confirmed by thermopower measurements. Doping requires an increase in Ga flux during growth resulting in a mixed phase material of polycrystalline GaAs:N embedded in amorphous GaN{sub 1-x}As{sub x}.

  15. Skin-effect down hole electric heater for heavy oil and high wax content oil applications

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, Chenglin; Wang, Hui; Liu, Yanhua [Liaoning Huafu Petroleum High-Tech Co., Ltd. (China); Xiao, Jon H; Klotz, Eric [ANDMIR Environmental Group Canada Inc. (Canada)

    2011-07-01

    With the increased production of oil and the depletion of conventional reserves, operators have started to exploit heavy oil and high wax content oil. Adequate production of such oils is difficult to achieve due to viscosity increase and mobility decrease during lifting as a result of heat loss. The down-hole electric heater has been developed to resolve these issues with the application of skin-effect electric heating technology. The aim of this paper is to present how this technology improves the production of heavy oil and waxy oil. Applications of the technology to wells in Chinese oilfields are studied. Results proved the technology to be efficient while being based on a simple process and operating in an easy and safe manner. This paper showed that the down-hole electric heater is a breakthrough technology, resolving the issues encountered in the heavy oil and waxy oil exploitation field, with broad application prospects.

  16. Characterization of SPAD Array for Multifocal High-Content Screening Applications

    Directory of Open Access Journals (Sweden)

    Anthony Tsikouras

    2016-10-01

    Full Text Available Current instruments used to detect specific protein-protein interactions in live cells for applications in high-content screening (HCS are limited by the time required to measure the lifetime. Here, a 32 × 1 single-photon avalanche diode (SPAD array was explored as a detector for fluorescence lifetime imaging (FLIM in HCS. Device parameters and characterization results were interpreted in the context of the application to determine if the SPAD array could satisfy the requirements of HCS-FLIM. Fluorescence lifetime measurements were performed using a known fluorescence standard; and the recovered fluorescence lifetime matched literature reported values. The design of a theoretical 32 × 32 SPAD array was also considered as a detector for a multi-point confocal scanning microscope.

  17. Identifying Content Knowledge for Teaching Energy: Examples from High School Physics

    Science.gov (United States)

    Robertson, Amy D.; Scherr, Rachel E.; Goodhew, Lisa M.; Daane, Abigail R.; Gray, Kara E.; Aker, Leanna B.

    2017-01-01

    "Content knowledge for teaching" is the specialized content knowledge that teachers use in practice the content knowledge that serves them for tasks of teaching such as revoicing students' ideas, choosing an instructional activity to address a student misunderstanding, and evaluating student statements. We describe a methodology for…

  18. Identifying Content Knowledge for Teaching Energy: Examples from High School Physics

    Science.gov (United States)

    Robertson, Amy D.; Scherr, Rachel E.; Goodhew, Lisa M.; Daane, Abigail R.; Gray, Kara E.; Aker, Leanna B.

    2017-01-01

    "Content knowledge for teaching" is the specialized content knowledge that teachers use in practice the content knowledge that serves them for tasks of teaching such as revoicing students' ideas, choosing an instructional activity to address a student misunderstanding, and evaluating student statements. We describe a methodology for…

  19. Solar activity variability in the IRI at high latitudes: Comparisons with GPS total electron content

    Science.gov (United States)

    Themens, David R.; Jayachandran, P. T.

    2016-04-01

    Total electron content (TEC) measurements from 10 dual-frequency GPS receivers in the Canadian High Arctic Ionospheric Network (CHAIN) are used to evaluate the performance of International Reference Ionosphere (IRI)-2007 within the Canadian sector. Throughout the region, we see systematic underestimation of daytime TEC, particularly at solar maximum, where summer and equinox root-mean-square errors reach as high as 14 total electron content units, 1 TECU = 1016 el m-2 (TECU). It is also shown that the use of a monthly IG index, in place of the IRI's standard IG12 index, leads to an improvement in TEC specification by up to 3 TECU in the polar cap and up to 6 TECU in the subauroral region during periods of short-term, large amplitude changes in solar activity. On diurnal timescales, variability in TEC is found to be underestimated by the IRI, during equinox periods, by up to 40% at subauroral latitudes and up to 70% in the polar cap region. During the winter, diurnal variations are overestimated by up to 40% in the subauroral region and are underestimated within the polar cap by up to 80%. Using collocated ionosonde data, we find IRI bottomside TEC to be within 1 TECU of observation with errors largest during the equinoxes. For the topside we find good agreement during the winter but significant underestimation of topside TEC by the IRI during summer and equinox periods, exceeding 6 TECU at times. By ingesting measured NmF2 into the IRI, we show that the topside thickness parameterization is the source of the bulk of the observed TEC errors.

  20. High-throughput retrotransposon-based fluorescent markers: improved information content and allele discrimination

    Directory of Open Access Journals (Sweden)

    Baker David

    2009-07-01

    Full Text Available Abstract Background Dense genetic maps, together with the efficiency and accuracy of their construction, are integral to genetic studies and marker assisted selection for plant breeding. High-throughput multiplex markers that are robust and reproducible can contribute to both efficiency and accuracy. Multiplex markers are often dominant and so have low information content, this coupled with the pressure to find alternatives to radio-labelling, has led us to adapt the SSAP (sequence specific amplified polymorphism marker method from a 33P labelling procedure to fluorescently tagged markers analysed from an automated ABI 3730 xl platform. This method is illustrated for multiplexed SSAP markers based on retrotransposon insertions of pea and is applicable for the rapid and efficient generation of markers from genomes where repetitive element sequence information is available for primer design. We cross-reference SSAP markers previously generated using the 33P manual PAGE system to fluorescent peaks, and use these high-throughput fluorescent SSAP markers for further genetic studies in Pisum. Results The optimal conditions for the fluorescent-labelling method used a triplex set of primers in the PCR. These included a fluorescently labelled specific primer together with its unlabelled counterpart, plus an adapter-based primer with two bases of selection on the 3' end. The introduction of the unlabelled specific primer helped to optimise the fluorescent signal across the range of fragment sizes expected, and eliminated the need for extensive dilutions of PCR amplicons. The software (GeneMarker Version 1.6 used for the high-throughput data analysis provided an assessment of amplicon size in nucleotides, peak areas and fluorescence intensity in a table format, so providing additional information content for each marker. The method has been tested in a small-scale study with 12 pea accessions resulting in 467 polymorphic fluorescent SSAP markers of which

  1. Terahertz-dependent evaluation of water content in high-water-cut crude oil using additive-manufactured samplers

    Science.gov (United States)

    Guan, LiMei; Zhan, HongLei; Miao, XinYang; Zhu, Jing; Zhao, Kun

    2017-04-01

    The evaluation of water content in crude oil is of significance to petroleum exploration and transportation. Terahertz (THz) waves are sensitive to fluctuations in the dipole moment of water. However, due to the strong absorption of water in the THz range, it is difficult for the THz spectrum to determine high water content with the common sampler. In this research, micron-grade samplers for THz detection were designed and manufactured using additive manufacturing (AM) technology. Oil-water mixtures with water content from 1.8% to 90.6% were measured with the THz-TDS system using sample cells. In addition, a detailed analysis was performed of the relationships among THz parameters such as signal peak, time delay, and refractive index as well as absorption coefficient and high water content (>60%). Results suggest that the combination of THz spectroscopy and AM technique is effective for water content evaluation in crude oil and can be further applied in the petroleum industry.

  2. Terahertz-dependent evaluation of water content in high-water-cut crude oil using additive-manufactured samplers

    Science.gov (United States)

    Guan, LiMei; Zhan, HongLei; Miao, XinYang; Zhu, Jing; Zhao, Kun

    2017-04-01

    The evaluation of water content in crude oil is of significance to petroleum exploration and transportation. Terahertz (THz) waves are sensitive to fluctuations in the dipole moment of water. However, due to the strong absorption of water in the THz range, it is difficult for the THz spectrum to determine high water content with the common sampler. In this research, micron-grade samplers for THz detection were designed and manufactured using additive manufacturing (AM) technology. Oil-water mixtures with water content from 1.8% to 90.6% were measured with the THz-TDS system using sample cells. In addition, a detailed analysis was performed of the relationships among THz parameters such as signal peak, time delay, and refractive index as well as absorption coefficient and high water content (>60%). Results suggest that the combination of THz spectroscopy and AM technique is effective for water content evaluation in crude oil and can be further applied in the petroleum industry.

  3. High temperature ultralow water content carbon dioxide-in-water foam stabilized with viscoelastic zwitterionic surfactants.

    Science.gov (United States)

    Alzobaidi, Shehab; Da, Chang; Tran, Vu; Prodanović, Maša; Johnston, Keith P

    2017-02-15

    Ultralow water content carbon dioxide-in-water (C/W) foams with gas phase volume fractions (ϕ) above 0.95 (that is foams may be stabilized with viscoelastic aqueous phases formed with a single zwitterionic surfactant at a concentration of only 1% (w/v) in DI water and over a wide range of salinity. Moreover, they are stable with a foam quality ϕ up to 0.98 even for temperatures up to 120°C. The properties of aqueous viscoelastic solutions and foams containing these solutions are examined for a series of zwitterionic amidopropylcarbobetaines, R-ONHC3H6N(CH3)2CH2CO2, where R is varied from C12-14 (coco) to C18 (oleyl) to C22 (erucyl). For the surfactants with long C18 and C22 tails, the relaxation times from complex rheology indicate the presence of viscoelastic wormlike micelles over a wide range in salinity and pH, given the high surfactant packing fraction. The apparent viscosities of these ultralow water content foams reached more than 120cP with stabilities more than 30-fold over those for foams formed with the non-viscoelastic C12-14 surfactant. At 90°C, the foam morphology was composed of ∼35μm diameter bubbles with a polyhedral texture. The apparent foam viscosity typically increased with ϕ and then dropped at ϕ values higher than 0.95-0.98. The Ostwald ripening rate was slower for foams with viscoelastic versus non-viscoelastic lamellae as shown by optical microscopy, as a consequence of slower lamellar drainage rates. The ability to achieve high stabilities for ultralow water content C/W foams over a wide temperature range is of interest in various technologies including polymer and materials science, CO2 enhanced oil recovery, CO2 sequestration (by greater control of the CO2 flow patterns), and possibly even hydraulic fracturing with minimal use of water to reduce the requirements for wastewater disposal.

  4. Spatial variability in the soil water content of a Mediterranean agroforestry system with high soil heterogeneity

    Science.gov (United States)

    Molina, Antonio Jaime; Llorens, Pilar; Aranda, Xavier; Savé, Robert; Biel, Carmen

    2013-04-01

    Variability of soil water content is known to increase with the size of spatial domain in which measurements are taken. At field scale, heterogeneity in soil, vegetation, topography, water input volume and management affects, among other factors, hydrologic plot behaviour under different mean soil water contents. The present work studies how the spatial variability of soil water content (SWC) is affected by soil type (texture, percentage of stones and the combination of them) in a timber-orientated plantation of cherry tree (Prunus avium) under Mediterranean climatic conditions. The experimental design is a randomized block one with 3 blocks * 4 treatments, based on two factors: irrigation (6 plots irrigated versus 6 plots not irrigated) and soil management (6 plots tillaged versus 6 plots not tillaged). SWC is continuously measured at 25, 50 and 100 cm depth with FDR sensors, located at two positions in each treatment: under tree influence and 2.5 m apart. This study presents the results of the monitoring during 2012 of the 24 sensors located at the 25 cm depth. In each of the measurement point, texture and percentage of stones were measured. Sandy-loam, sandy-clay-loam and loam textures were found together with a percentage of stones ranging from 20 to 70 %. The results indicated that the relationship between the daily mean SWC and its standard deviation, a common procedure used to study spatial variability, changed with texture, percentage of stones and the estimation of field capacity from the combination of both. Temporal stability analysis of SWC showed a clear pattern related to field capacity, with the measurement points of the sandy-loam texture and the high percentage of stones showing the maximun negative diference with the global mean. The high range in the mean relative difference observed (± 75 %), could indicate that the studied plot may be considered as a good field-laboratory to extrapolate results at higher spatial scales. Furthermore, the

  5. Cultivation Methods for Tomato (Lycopersicon esculentum Mill.) with High Soluble Solids Content Using the Vertically Split Root System

    National Research Council Canada - National Science Library

    HAYASHI, Hiroyuki; TOYOFUKU, Kyoko; TAGUCHI, Takiko; OGAWA, Atsushi

    2014-01-01

    For the cultivation of tomatoes with high soluble solids content, the root system was split vertically and irrigation to the upper part of the root system was stopped to create split vertically (SR treatment...

  6. BioSig3D: High Content Screening of Three-Dimensional Cell Culture Models: e0148379

    National Research Council Canada - National Science Library

    Cemal Cagatay Bilgin; Gerald Fontenay; Qingsu Cheng; Hang Chang; Ju Han; Bahram Parvin

    2016-01-01

    ...) cell culture models that are imaged in full 3D volume. It provides an end-to-end solution for designing high content screening assays, based on colony organization that is derived from segmentation of nuclei in each colony...

  7. High nitrogen supply and carbohydrate content reduce fungal endophyte and alkaloid concentration in Lolium perenne.

    Science.gov (United States)

    Rasmussen, Susanne; Parsons, Anthony J; Bassett, Shalome; Christensen, Michael J; Hume, David E; Johnson, Linda J; Johnson, Richard D; Simpson, Wayne R; Stacke, Christina; Voisey, Christine R; Xue, Hong; Newman, Jonathan A

    2007-01-01

    The relationship between cool-season grasses and fungal endophytes is widely regarded as mutualistic, but there is growing uncertainty about whether changes in resource supply and environment benefit both organisms to a similar extent. Here, we infected two perennial ryegrass (Lolium perenne) cultivars (AberDove, Fennema) that differ in carbohydrate content with three strains of Neotyphodium lolii (AR1, AR37, common strain) that differ intrinsically in alkaloid profile. We grew endophyte-free and infected plants under high and low nitrogen (N) supply and used quantitative PCR (qPCR) to estimate endophyte concentrations in harvested leaf tissues. Endophyte concentration was reduced by 40% under high N supply, and by 50% in the higher sugar cultivar. These two effects were additive (together resulting in 75% reduction). Alkaloid production was also reduced under both increased N supply and high sugar cultivar, and for three of the four alkaloids quantified, concentrations were linearly related to endophyte concentration. The results stress the need for wider quantification of fungal endophytes in the grassland-foliar endophyte context, and have implications for how introducing new cultivars, novel endophytes or increasing N inputs affect the role of endophytes in grassland ecosystems.

  8. Halophilic properties of metal binding protein characterized by high histidine content from Chromohalobacter salexigens DSM3043.

    Science.gov (United States)

    Yamaguchi, Rui; Arakawa, Tsutomu; Tokunaga, Hiroko; Ishibashi, Matsujiro; Tokunaga, Masao

    2012-02-01

    Periplasmic metal binding protein characterized by high histidine content was cloned from moderate halophile, Chromohalobacter salexigens. The protein, termed histidine-rich metal binding protein (HP), was expressed in and purified from E. coli as a native form. HP bound to Ni- and Cu-loaded chelate columns with high affinity, and Co- and Zn-columns with moderate affinity. Although the secondary structure was not grossly altered by the addition of 0.2-2.0 M NaCl, the thermal transition pattern was considerably shifted to higher temperature with increasing salt concentration: melting temperature was raised by ~20 °C at 2.0 M NaCl over the melting temperature at 0.2 M NaCl. HP showed reversible refolding from thermal melting in 0.2-1.15 M NaCl, while it formed irreversible aggregates upon thermal melting at 2 M NaCl. Addition of 0.01-0.1 mM NiSO₄ stabilized HP against thermal melting with high reversibility, while addition above 0.5 mM resulted in irreversible melting due to aggregation.

  9. Development and freeze-thaw durability of high flyash-content concrete

    Energy Technology Data Exchange (ETDEWEB)

    Sajadi, J.

    1987-01-01

    Objectives were to investigate the effects on concrete strength, drying shrinkage, freeze-thaw durability, and air-void system parameters of replacing various amounts of portland cement with different types of fly ash and to compare selected characteristics of such fly-ash concretes and fly-ash concretes containing a high-range water-reducing admixture to those of a control mixture. It was concluded that concrete mixtures with 90-day compressive strengths equal to the control could be produced when large amounts of cement were replaced by fly ash. In addition, when the high-range water-reducing admixtures was employed, very large amounts of cement could be replaced by fly ash to yield mixtures whose compressive strengths were equal to or greater than the strengths of the control mix at all ages. The maximum amount of cement that could be replaced for equal-strength mixtures depended upon the nature of the fly ash. Drying shrinkage of plain fly-ash concretes and fly-ash concretes containing the high-range water-reducing admixture were similar to those of the control mix. The optimum fly-ash content in a concrete is comparable in strength and durability to a conventional (control) concrete was influenced by the chemical and physical characteristics of the fly ash.

  10. Evaluation of Mechanical Properties According to Nb Content of High Strength Steel Manufactured in VIM

    Institute of Scientific and Technical Information of China (English)

    Byung-Hui Choi; Byung-Ki Choi

    2008-01-01

    As the aircraft and aerospace industry and the automobile industry are developed, the demand of reliable materials with high strength and high toughness is steadily increased. The grain miniature method which improves the ductility and the toughness simultaneously under high strength is the miniature of the casting microstructure. In this paper, the Nb which affects the austenite grain miniature is added by 0.00%, 0.03% and 0.06% in this steel and the ingot is manufactured in a vacuum induction melting furnace (VIM). The castingmi crostructure and the mechanical property of the maraging steel according to Nb contents are analyzed by conducting the solution annealing and the age hardening after hot rolling. In this result, a specimen containing 0.03% Nb is most miniature. When it does the long time age hardening, the precipitation estimated as the carbide is appeared. The mechanical properties are excellent if a specimen containing 0.06% Nb does the age hardening during 8 h in 482℃.

  11. Predicting In Vivo Anti-Hepatofibrotic Drug Efficacy Based on In Vitro High-Content Analysis

    Science.gov (United States)

    Zheng, Baixue; Tan, Looling; Mo, Xuejun; Yu, Weimiao; Wang, Yan; Tucker-Kellogg, Lisa; Welsch, Roy E.; So, Peter T. C.; Yu, Hanry

    2011-01-01

    Background/Aims Many anti-fibrotic drugs with high in vitro efficacies fail to produce significant effects in vivo. The aim of this work is to use a statistical approach to design a numerical predictor that correlates better with in vivo outcomes. Methods High-content analysis (HCA) was performed with 49 drugs on hepatic stellate cells (HSCs) LX-2 stained with 10 fibrotic markers. ∼0.3 billion feature values from all cells in >150,000 images were quantified to reflect the drug effects. A systematic literature search on the in vivo effects of all 49 drugs on hepatofibrotic rats yields 28 papers with histological scores. The in vivo and in vitro datasets were used to compute a single efficacy predictor (Epredict). Results We used in vivo data from one context (CCl4 rats with drug treatments) to optimize the computation of Epredict. This optimized relationship was independently validated using in vivo data from two different contexts (treatment of DMN rats and prevention of CCl4 induction). A linear in vitro-in vivo correlation was consistently observed in all the three contexts. We used Epredict values to cluster drugs according to efficacy; and found that high-efficacy drugs tended to target proliferation, apoptosis and contractility of HSCs. Conclusions The Epredict statistic, based on a prioritized combination of in vitro features, provides a better correlation between in vitro and in vivo drug response than any of the traditional in vitro markers considered. PMID:22073152

  12. Quantification of histone H3 Lys27 trimethylation (H3K27me3) by high-throughput microscopy enables cellular large-scale screening for small-molecule EZH2 inhibitors.

    Science.gov (United States)

    Luense, Svenja; Denner, Philip; Fernández-Montalván, Amaury; Hartung, Ingo; Husemann, Manfred; Stresemann, Carlo; Prechtl, Stefan

    2015-02-01

    EZH2 inhibition can decrease global histone H3 lysine 27 trimethylation (H3K27me3) and thereby reactivates silenced tumor suppressor genes. Inhibition of EZH2 is regarded as an option for therapeutic cancer intervention. To identify novel small-molecule (SMOL) inhibitors of EZH2 in drug discovery, trustworthy cellular assays amenable for phenotypic high-throughput screening (HTS) are crucial. We describe a reliable approach that quantifies changes in global levels of histone modification marks using high-content analysis (HCA). The approach was validated in different cell lines by using small interfering RNA and SMOL inhibitors. By automation and miniaturization from a 384-well to 1536-well plate, we demonstrated its utility in conducting phenotypic HTS campaigns and assessing structure-activity relationships (SAR). This assay enables screening of SMOL EZH2 inhibitors and can advance the mechanistic understanding of H3K27me3 suppression, which is crucial with regard to epigenetic therapy. We observed that a decrease in global H3K27me3, induced by EZH2 inhibition, comprises two distinct mechanisms: (1) inhibition of de novo DNA methylation and (II) inhibition of dynamic, replication-independent H3K27me3 turnover. This report describes an HCA assay for primary HTS to identify, profile, and optimize cellular active SMOL inhibitors targeting histone methyltransferases, which could benefit epigenetic drug discovery. © 2014 Society for Laboratory Automation and Screening.

  13. Solution-Processable Organic Molecule for High-Performance Organic Solar Cells with Low Acceptor Content.

    Science.gov (United States)

    Wang, Kun; Guo, Bing; Xu, Zhuo; Guo, Xia; Zhang, Maojie; Li, Yongfang

    2015-11-11

    A new planar D2-A-D1-A-D2 structured organic molecule with bithienyl benzodithiophene (BDT) as central donor unit D1 and fluorine-substituted benzothiadiazole (BTF) as acceptor unit and alkyl-dithiophene as end group and donor unit D2, BDT-BTF, was designed and synthesized for the application as donor material in organic solar cells (OSCs). BDT-BTF shows a broad absorption in visible region, suitable highest occupied molecular orbital energy level of -5.20 eV, and high hole mobility of 1.07 × 10(-2) cm(2)/(V s), benefitted from its high coplanarity and strong crystallinity. The OSCs based on BDT-BTF as donor (D) and PC71BM as acceptor (A) at a D/A weight ratio of 3:1 without any extra treatment exhibit high photovoltaic performance with Voc of 0.85 V, Jsc of 10.48 mA/cm(2), FF of 0.66, and PCE of 5.88%. The morphological study by transmission electron microscopy reveals that the blend of BDT-BTF and PC71BM (3:1, w/w) possesses an appropriate interpenetrating D/A network for the exciton separation and charge carrier transport, which agrees well with the good device performance. The optimized D/A weight ratio of 3:1 is the lowest acceptor content in the active layer reported so far for the high-performance OSCs, and the organic molecules with the molecular structure like BDT-BTF could be promising high-performance donor materials in solution-processable OSCs.

  14. [Effects of high temperature on Bt protein content and nitrogen metabolic physiology in boll wall of Bt cotton].

    Science.gov (United States)

    Wang, Jun; Abidallah, Eltayib H M A; Hua, Ming-ming; Heng, Li; Lyu, Chun-hua; Chen, De-hua

    2015-10-01

    Bt cotton cultivar Sikang 1 (a conventional cultivar) and Sikang 3 (a hybrid cultivar) from China, and 99B (a conventional cultivar) and Daiza 1 (a hybrid cultivar) from USA were selected as experimental materials, the ball wall Bt protein content and nitrogen metabolic physiology were investigated under different high temperature levels at peak boll stage. The results showed that the Bt protein content of boll wall decreased with the increasing temperature. Compared with the control (32 °C, the boll wall Bt protein content decreased significantly when the temperature was above 38 °C for the conventional cultivars and above 40 °C for the hybrid cultivars. The Bt protein contents of cultivar Sikang 1 and 99B decreased by 53.0% and 69.5% respectively with the temperature at 38 °C, and that of cultivar Sikang 3 and Daiza 1 decreased by 64.8% and 54.1% respectively with the temperature at 40 °C. Greater reductions in the boll wall soluble protein contents and GPT activities, larger increments for the boll wall free amino acid contents and proteinsase activities were also observed when the boll wall Bt protein content was significantly reduced. Therefore, high temperature resulted in the reduction of Bt protein synthesis and increase of the insecticidal protein degradation in the boll wall significantly, which caused the reductions in boll wall Bt protein content and insect resistance.

  15. Wild plants as tools for the remediation of abandoned mining sites with a high arsenic content

    Science.gov (United States)

    Martínez-Lopez, Salvadora; Martínez-Sanchez, MJose; Perez-Sirvent, Carmen; Martínez, Lucia B.; Bech, Jaume

    2014-05-01

    The aim of this study was to assess the environmental risk posed by arsenic when new vegetation types are introduced, analyzing the transfer of arsenic in different plant species that grow spontaneously in mining areas of SE Spain (Sierra Minera of Cartagena), and the contribution of such plants to the environmental risk represented by their ingestion by animals living in the same ecosystems. When dealing with remediation projects in zones affected by mining activities, the risk posed by the ingestion of the plants by fauna is often forgotten. To study the transfer to the trophic chain, two mammals, sheep and vole, were selected. The risk analysis was centered in the contribution of these natural plants to the ingestion calculated. For this study, 21 vegetal species naturally growing in the soils were collected from the Sierra Minera. The vegetal material studied is clearly associated with the Mediterranean Region (S.E. of Spain) and the plant species collected are endemisms and plants characteristic of the zone. Physico-chemical properties were obtained by means of the usual procedures. To determine the arsenic content, the soil samples and plant materials were digested in a microwave system and the arsenic concentration was determined using atomic fluorescence spectrometry with an automated continuous flow hydride generation system. A semiquantitative estimation of the mineralogical composition of the samples was made by X Ray Diffraction analysis. The soils were classified into three groups: Low (group 1) (7-35 mg/kg) medium (group 2) (35-327 mg/kg) and high (group 3) (> 327 mg/kg), according to their As content. The mineralogy and As content of the soils studied depends on the materials related with mining activity. The descriptive statistical analysis of the population of plants studied showed the As range in roots to be 0.31-150 mg/kg while leaf concentrations were lower (0.21-83.4 mg/kg). The potential risk of As entering the food chain through of the plant

  16. Aromatic claw: A new fold with high aromatic content that evades structural prediction.

    Science.gov (United States)

    Sachleben, Joseph R; Adhikari, Aashish N; Gawlak, Grzegorz; Hoey, Robert J; Liu, Gaohua; Joachimiak, Andrzej; Montelione, Gaetano T; Sosnick, Tobin R; Koide, Shohei

    2017-02-01

    We determined the NMR structure of a highly aromatic (13%) protein of unknown function, Aq1974 from Aquifex aeolicus (PDB ID: 5SYQ). The unusual sequence of this protein has a tryptophan content five times the normal (six tryptophan residues of 114 or 5.2% while the average tryptophan content is 1.0%) with the tryptophans occurring in a WXW motif. It has no detectable sequence homology with known protein structures. Although its NMR spectrum suggested that the protein was rich in β-sheet, upon resonance assignment and solution structure determination, the protein was found to be primarily α-helical with a small two-stranded β-sheet with a novel fold that we have termed an Aromatic Claw. As this fold was previously unknown and the sequence unique, we submitted the sequence to CASP10 as a target for blind structural prediction. At the end of the competition, the sequence was classified a hard template based model; the structural relationship between the template and the experimental structure was small and the predictions all failed to predict the structure. CSRosetta was found to predict the secondary structure and its packing; however, it was found that there was little correlation between CSRosetta score and the RMSD between the CSRosetta structure and the NMR determined one. This work demonstrates that even in relatively small proteins, we do not yet have the capacity to accurately predict the fold for all primary sequences. The experimental discovery of new folds helps guide the improvement of structural prediction methods.

  17. The high-ion content and kinematics of low-redshift Lyman limit systems

    Energy Technology Data Exchange (ETDEWEB)

    Fox, Andrew J.; Tumlinson, Jason; Bordoloi, Rongmon [Space Telescope Science Institute, 3700 San Martin Drive, Baltimore, MD 21218 (United States); Lehner, Nicolas; Howk, J. Christopher [Department of Physics, University of Notre Dame, 225 Nieuwland Science Hall, Notre Dame, IN 46556 (United States); Tripp, Todd M.; Katz, Neal [Department of Astronomy, University of Massachusetts, Amherst, MA 01003 (United States); Prochaska, J. Xavier; Werk, Jessica K. [UCO/Lick Observatory, University of California, Santa Cruz, CA 95064 (United States); O' Meara, John M. [Department of Physics, Saint Michael' s College, One Winooski Park, Colchester, VT 05439 (United States); Oppenheimer, Benjamin D. [Leiden Observatory, Leiden University, NL-2300 RA Leiden (Netherlands); Davé, Romeel, E-mail: afox@stsci.edu [University of the Western Cape, Robert Sobukwe Road, Bellville 7535 (South Africa)

    2013-12-01

    We study the high-ion content and kinematics of the circumgalactic medium around low-redshift galaxies using a sample of 23 Lyman limit systems (LLSs) at 0.08 < z < 0.93 observed with the Cosmic Origins Spectrograph on board the Hubble Space Telescope. In Lehner et al., we recently showed that low-z LLSs have a bimodal metallicity distribution. Here we extend that analysis to search for differences between the high-ion and kinematic properties of the metal-poor and metal-rich branches. We find that metal-rich LLSs tend to show higher O VI columns and broader O VI profiles than metal-poor LLSs. The total H I line width (Δv {sub 90} statistic) in LLSs is not correlated with metallicity, indicating that the H I kinematics alone cannot be used to distinguish inflow from outflow and gas recycling. Among the 17 LLSs with O VI detections, all but two show evidence of kinematic sub-structure, in the form of O VI-H I centroid offsets, multiple components, or both. Using various scenarios for how the metallicities in the high-ion and low-ion phases of each LLS compare, we constrain the ionized hydrogen column in the O VI phase to lie in the range log N(H II) ∼ 17.6-20. The O VI phase of LLSs is a substantial baryon reservoir, with M(high-ion) ∼ 10{sup 8.5-10.9} (r/150 kpc){sup 2} M {sub ☉}, similar to the mass in the low-ion phase. Accounting for the O VI phase approximately doubles the contribution of low-z LLSs to the cosmic baryon budget.

  18. Evidence for high fluid/melt content beneath Krakatau volcano (Indonesia) from local earthquake tomography

    Science.gov (United States)

    Jaxybulatov, Kairly; Koulakov, Ivan; Seht, Malte Ibs-von; Klinge, Klaus; Reichert, Christian; Dahren, Börje; Troll, Valentin R.

    2011-09-01

    Within the KRAKMON project for multiparameter monitoring of Anak Krakatau volcano (Indonesia), a network of temporary stations was installed on the islands of the Krakatau complex as well as in the surrounding areas of the Sunda Strait, Sumatra and Java. The network was operated from June 2005 until January 2006. More than 700 local events were recorded during this experiment, and travel times from these events were used to perform a tomographic inversion for P and S velocities and for the Vp/Vs ratio. In this study, special attention was paid to the validation of the computed model based on different tests, such as inversion of independent data subsets and synthetic modeling. Although the network configuration and the distribution of the events are not favorable for high-quality tomographic imaging, we have obtained some important and robust features which give information about sources of volcanic activity in the Krakatau complex. The most interesting feature of this study is a zone of high Vp/Vs ratio beneath the Krakatau complex. At depths down to 4 km depth we observe anticorrelation of higher P- and lower S-velocities that leads to Vp/Vs ratio higher than 2. This is a probable indicator of the presence of partially molten and/or with high fluid content material with a composition corresponding to deeper layers. It is important that the anomaly of high Vp/Vs ratio beneath the Krakatau complex appears to be separated in two parts at a depth of 5-6 km. This fits to results of geobarometric analysis that presume the existence of several levels of magma chambers beneath Anak Krakatau.

  19. Effect of Simulated High Hydrogen Content Combustion Environments on Abradable Properties of Ceramic Turbine Coatings

    Science.gov (United States)

    Basu Majumder, Madhura

    Air plasma sprayed (APS) abradable coatings are used in the turbine hot section to reduce the stator-rotor gap, minimizing gas leakage. These coatings are designed to exhibit controlled removal of material in thin layers when the turbine blades sweep through the coating, which protects the mechanical integrity of the turbine blade. In an effort to lower CO2 emissions, high H2 content fuel is being explored. This change in chemical composition of the fuel may affect the microstructure, abradability and durability of the coatings at turbine operational temperatures. The presence of high water vapor in the combustion chamber leads to accelerated degradation of the sacrificial coating materials. In this work, zirconia based composite materials with a machinable phase and varied porosity have been used to study microstructural evolution, thermal and chemical stability of the phases and abradable characteristics of baseline coating systems in both humid and dry environments. Investigation of the mechanisms that control the removal of materials and performance of abradable coatings through thermo-mechanical tests will be discussed.

  20. Teachable, high-content analytics for live-cell, phase contrast movies.

    Science.gov (United States)

    Alworth, Samuel V; Watanabe, Hirotada; Lee, James S J

    2010-09-01

    CL-Quant is a new solution platform for broad, high-content, live-cell image analysis. Powered by novel machine learning technologies and teach-by-example interfaces, CL-Quant provides a platform for the rapid development and application of scalable, high-performance, and fully automated analytics for a broad range of live-cell microscopy imaging applications, including label-free phase contrast imaging. The authors used CL-Quant to teach off-the-shelf universal analytics, called standard recipes, for cell proliferation, wound healing, cell counting, and cell motility assays using phase contrast movies collected on the BioStation CT and BioStation IM platforms. Similar to application modules, standard recipes are intended to work robustly across a wide range of imaging conditions without requiring customization by the end user. The authors validated the performance of the standard recipes by comparing their performance with truth created manually, or by custom analytics optimized for each individual movie (and therefore yielding the best possible result for the image), and validated by independent review. The validation data show that the standard recipes' performance is comparable with the validated truth with low variation. The data validate that the CL-Quant standard recipes can provide robust results without customization for live-cell assays in broad cell types and laboratory settings.

  1. Designs and concept reliance of a fully automated high-content screening platform.

    Science.gov (United States)

    Radu, Constantin; Adrar, Hosna Sana; Alamir, Ab; Hatherley, Ian; Trinh, Trung; Djaballah, Hakim

    2012-10-01

    High-content screening (HCS) is becoming an accepted platform in academic and industry screening labs and does require slightly different logistics for execution. To automate our stand-alone HCS microscopes, namely, an alpha IN Cell Analyzer 3000 (INCA3000), originally a Praelux unit hooked to a Hudson Plate Crane with a maximum capacity of 50 plates per run, and the IN Cell Analyzer 2000 (INCA2000), in which up to 320 plates could be fed per run using the Thermo Fisher Scientific Orbitor, we opted for a 4 m linear track system harboring both microscopes, plate washer, bulk dispensers, and a high-capacity incubator allowing us to perform both live and fixed cell-based assays while accessing both microscopes on deck. Considerations in design were given to the integration of the alpha INCA3000, a new gripper concept to access the onboard nest, and peripheral locations on deck to ensure a self-reliant system capable of achieving higher throughput. The resulting system, referred to as Hestia, has been fully operational since the new year, has an onboard capacity of 504 plates, and harbors the only fully automated alpha INCA3000 unit in the world.

  2. Ethanol production from food waste at high solids content with vacuum recovery technology.

    Science.gov (United States)

    Huang, Haibo; Qureshi, Nasib; Chen, Ming-Hsu; Liu, Wei; Singh, Vijay

    2015-03-18

    Ethanol production from food wastes does not only solve environmental issues but also provides renewable biofuels. This study investigated the feasibility of producing ethanol from food wastes at high solids content (35%, w/w). A vacuum recovery system was developed and applied to remove ethanol from fermentation broth to reduce yeast ethanol inhibition. A high concentration of ethanol (144 g/L) was produced by the conventional fermentation of food waste without a vacuum recovery system. When the vacuum recovery is applied to the fermentation process, the ethanol concentration in the fermentation broth was controlled below 100 g/L, thus reducing yeast ethanol inhibition. At the end of the conventional fermentation, the residual glucose in the fermentation broth was 5.7 g/L, indicating incomplete utilization of glucose, while the vacuum fermentation allowed for complete utilization of glucose. The ethanol yield for the vacuum fermentation was found to be 358 g/kg of food waste (dry basis), higher than that for the conventional fermentation at 327 g/kg of food waste (dry basis).

  3. Laser-based single-axon transection for high-content axon injury and regeneration studies.

    Directory of Open Access Journals (Sweden)

    Darío Kunik

    Full Text Available The investigation of the regenerative response of the neurons to axonal injury is essential to the development of new axoprotective therapies. Here we study the retinal neuronal RGC-5 cell line after laser transection, demonstrating that the ability of these cells to initiate a regenerative response correlates with axon length and cell motility after injury. We show that low energy picosecond laser pulses can achieve transection of unlabeled single axons in vitro and precisely induce damage with micron precision. We established the conditions to achieve axon transection, and characterized RGC-5 axon regeneration and cell body response using time-lapse microscopy. We developed an algorithm to analyze cell trajectories and established correlations between cell motility after injury, axon length, and the initiation of the regeneration response. The characterization of the motile response of axotomized RGC-5 cells showed that cells that were capable of repair or regrowth of damaged axons migrated more slowly than cells that could not. Moreover, we established that RGC-5 cells with long axons could not recover their injured axons, and such cells were much more motile. The platform we describe allows highly controlled axonal damage with subcellular resolution and the performance of high-content screening in cell cultures.

  4. High-Content Analysis of Breast Cancer Using Single-Cell Deep Transfer Learning.

    Science.gov (United States)

    Kandaswamy, Chetak; Silva, Luís M; Alexandre, Luís A; Santos, Jorge M

    2016-03-01

    High-content analysis has revolutionized cancer drug discovery by identifying substances that alter the phenotype of a cell, which prevents tumor growth and metastasis. The high-resolution biofluorescence images from assays allow precise quantitative measures enabling the distinction of small molecules of a host cell from a tumor. In this work, we are particularly interested in the application of deep neural networks (DNNs), a cutting-edge machine learning method, to the classification of compounds in chemical mechanisms of action (MOAs). Compound classification has been performed using image-based profiling methods sometimes combined with feature reduction methods such as principal component analysis or factor analysis. In this article, we map the input features of each cell to a particular MOA class without using any treatment-level profiles or feature reduction methods. To the best of our knowledge, this is the first application of DNN in this domain, leveraging single-cell information. Furthermore, we use deep transfer learning (DTL) to alleviate the intensive and computational demanding effort of searching the huge parameter's space of a DNN. Results show that using this approach, we obtain a 30% speedup and a 2% accuracy improvement.

  5. High-content analysis to leverage a robust phenotypic profiling approach to vascular modulation.

    Science.gov (United States)

    Isherwood, Beverley J; Walls, Rebecca E; Roberts, Mark E; Houslay, Thomas M; Brave, Sandra R; Barry, Simon T; Carragher, Neil O

    2013-12-01

    Phenotypic screening seeks to identify substances that modulate phenotypes in a desired manner with the aim of progressing first-in-class agents. Successful campaigns require physiological relevance, robust screening, and an ability to deconvolute perturbed pathways. High-content analysis (HCA) is increasingly used in cell biology and offers one approach to prosecution of phenotypic screens, but challenges exist in exploitation where data generated are high volume and complex. We combine development of an organotypic model with novel HCA tools to map phenotypic responses to pharmacological perturbations. We describe implementation for angiogenesis, a process that has long been a focus for therapeutic intervention but has lacked robust models that recapitulate more completely mechanisms involved. The study used human primary endothelial cells in co-culture with stromal fibroblasts to model multiple aspects of angiogenic signaling: cell interactions, proliferation, migration, and differentiation. Multiple quantitative descriptors were derived from automated microscopy using custom-designed algorithms. Data were extracted using a bespoke informatics platform that integrates processing, statistics, and feature display into a streamlined workflow for building and interrogating fingerprints. Ninety compounds were characterized, defining mode of action by phenotype. Our approach for assessing phenotypic outcomes in complex assay models is robust and capable of supporting a range of phenotypic screens at scale.

  6. Combination of Sardine and Shark Oil High Content of Omega-3 and Squalene

    Directory of Open Access Journals (Sweden)

    Muhamad Musbah

    2017-05-01

    Full Text Available Sardine oil contain high concentration of  EPA but low of  DHA while shark is reverse. Shark oil  high contain of DHA and squalene but low EPA. This research aim to fortify the quality of  fish oil withomega-3 and squalen and improve the quality of fish oil. The combination of fish oil (sardine:shark 1: 1, 1: 2, 1: 3, 1: 4, 2: 1, 3: 1 and 4:1 showed significant results on peroxide, anisidine, and total oxidation value, however free fatty acids analysis did not show the influence to the content value.  The best oxidation parameters value werefound (sardine: shark (1:4 with peroxide was 5.44±0.06 mEq/kg, anisidine was 8.3±0.72 mEq/kg and total oxidation was 19.27±0.7mEq/kg. Fatty acids profile between  sardines and shark oil containedvarious SFA, MUFA and PUFA. Sardine oil which was added more to combination ratio will increase omega-3. Sample 1:4 had 43.16% squalene.

  7. GPS scintillations and total electron content climatology in the southern low, middle and high latitude regions

    Directory of Open Access Journals (Sweden)

    Luca Spogli

    2013-06-01

    Full Text Available In recent years, several groups have installed high-frequency sampling receivers in the southern middle and high latitude regions, to monitor ionospheric scintillations and the total electron content (TEC changes. Taking advantage of the archive of continuous and systematic observations of the ionosphere on L-band by means of signals from the Global Positioning System (GPS, we present the first attempt at ionospheric scintillation and TEC mapping from Latin America to Antarctica. The climatology of the area considered is derived through Ground-Based Scintillation Climatology, a method that can identify ionospheric sectors in which scintillations are more likely to occur. This study also introduces the novel ionospheric scintillation 'hot-spot' analysis. This analysis first identifies the crucial areas of the ionosphere in terms of enhanced probability of scintillation occurrence, and then it studies the seasonal variation of the main scintillation and TEC-related parameters. The results produced by this sophisticated analysis give significant indications of the spatial/ temporal recurrences of plasma irregularities, which contributes to the extending of current knowledge of the mechanisms that cause scintillations, and consequently to the development of efficient tools to forecast space-weather-related ionospheric events.

  8. Activated carbon fibers with a high heteroatom content by chemical activation of PBO with phosphoric acid.

    Science.gov (United States)

    Vázquez-Santos, M B; Suárez-García, F; Martínez-Alonso, A; Tascón, J M D

    2012-04-03

    The preparation of activated carbon fibers (ACFs) by phosphoric acid activation of poly(p-phenylene benzobisoxazole) (PBO) fibers was studied, with particular attention to the effects of impregnation ratio and carbonization temperature on porous texture. Phosphoric acid has a strong effect on PBO degradation, lowering the temperature range at which the decomposition takes place and changing the number of mass loss steps. Chemical analysis results indicated that activation with phosphoric acid increases the concentration of oxygenated surface groups; the resulting materials also exhibiting high nitrogen content. ACFs are obtained with extremely high yields; they have well-developed porosity restricted to the micropore and narrow mesopore range and with a significant concentration of phosphorus incorporated homogeneously in the form of functional groups. An increase in the impregnation ratio leads to increases in both pore volume and pore size, maximum values of surface area (1250 m(2)/g) and total pore volume (0.67 cm(3)/g) being attained at the highest impregnation ratio (210 wt % H(3)PO(4)) and lowest activation temperature (650 °C) used; the corresponding yield was as large as 83 wt %. The obtained surface areas and pore volumes were higher than those achieved in previous works by physical activation with CO(2) of PBO chars.

  9. High-content live cell imaging with RNA probes: advancements in high-throughput antimalarial drug discovery

    Directory of Open Access Journals (Sweden)

    Cervantes Serena

    2009-06-01

    Full Text Available Abstract Background Malaria, a major public health issue in developing nations, is responsible for more than one million deaths a year. The most lethal species, Plasmodium falciparum, causes up to 90% of fatalities. Drug resistant strains to common therapies have emerged worldwide and recent artemisinin-based combination therapy failures hasten the need for new antimalarial drugs. Discovering novel compounds to be used as antimalarials is expedited by the use of a high-throughput screen (HTS to detect parasite growth and proliferation. Fluorescent dyes that bind to DNA have replaced expensive traditional radioisotope incorporation for HTS growth assays, but do not give additional information regarding the parasite stage affected by the drug and a better indication of the drug's mode of action. Live cell imaging with RNA dyes, which correlates with cell growth and proliferation, has been limited by the availability of successful commercial dyes. Results After screening a library of newly synthesized stryrl dyes, we discovered three RNA binding dyes that provide morphological details of live parasites. Utilizing an inverted confocal imaging platform, live cell imaging of parasites increases parasite detection, improves the spatial and temporal resolution of the parasite under drug treatments, and can resolve morphological changes in individual cells. Conclusion This simple one-step technique is suitable for automation in a microplate format for novel antimalarial compound HTS. We have developed a new P. falciparum RNA high-content imaging growth inhibition assay that is robust with time and energy efficiency.

  10. High Cellular Monocyte Activation in People Living With Human Immunodeficiency Virus on Combination Antiretroviral Therapy and Lifestyle-Matched Controls Is Associated With Greater Inflammation in Cerebrospinal Fluid

    Science.gov (United States)

    Booiman, Thijs; Wit, Ferdinand W.; Maurer, Irma; De Francesco, Davide; Sabin, Caroline A.; Harskamp, Agnes M.; Prins, Maria; Garagnani, Paolo; Pirazzini, Chiara; Franceschi, Claudio; Fuchs, Dietmar; Gisslén, Magnus; Winston, Alan; Reiss, Peter; Reiss, P.; Wit, F. W. N. M.; Schouten, J.; Kooij, K. W.; van Zoest, R. A.; Elsenga, B. C.; Janssen, F. R.; Heidenrijk, M.; Zikkenheiner, W.; van der Valk, M.; Kootstra, N. A.; Booiman, T.; Harskamp-Holwerda, A. M.; Boeser-Nunnink, B.; Maurer, I.; Mangas Ruiz, M. M.; Girigorie, A. F.; Villaudy, J.; Frankin, E.; Pasternak, A.; Berkhout, B.; van der Kuyl, T.; Portegies, P.; Schmand, B. A.; Geurtsen, G. J.; ter Stege, J. A.; Klein Twennaar, M.; Majoie, C. B. L. M.; Caan, M. W. A.; Su, T.; Weijer, K.; Bisschop, P. H. L. T.; Kalsbeek, A.; Wezel, M.; Visser, I.; Ruhé, H. G.; Franceschi, C.; Garagnani, P.; Pirazzini, C.; Capri, M.; Dall’Olio, F.; Chiricolo, M.; Salvioli, S.; Hoeijmakers, J.; Pothof, J.; Prins, M.; Martens, M.; Moll, S.; Berkel, J.; Totté, M.; Kovalev, S.; Gisslén, M.; Fuchs, D.; Zetterberg, H.; Winston, A.; Underwood, J.; McDonald, L.; Stott, M.; Legg, K.; Lovell, A.; Erlwein, O.; Doyle, N.; Kingsley, C.; Sharp, D. J.; Leech, R.; Cole, J. H.; Zaheri, S.; Hillebregt, M. M. J.; Ruijs, Y. M. C.; Benschop, D. P.; Burger, D.; de Graaff-Teulen, M.; Guaraldi, G.; Bürkle, A.; Sindlinger, T.; Moreno-Villanueva, M.; Keller, A.; Sabin, C.; de Francesco, D.; Libert, C.; Dewaele, S.

    2017-01-01

    Abstract Background. Increased monocyte activation and intestinal damage have been shown to be predictive for the increased morbidity and mortality observed in treated people living with human immunodeficiency virus (PLHIV). Methods. A cross-sectional analysis of cellular and soluble markers of monocyte activation, coagulation, intestinal damage, and inflammation in plasma and cerebrospinal fluid (CSF) of PLHIV with suppressed plasma viremia on combination antiretroviral therapy and age and demographically comparable HIV-negative individuals participating in the Comorbidity in Relation to AIDS (COBRA) cohort and, where appropriate, age-matched blood bank donors (BBD). Results. People living with HIV, HIV-negative individuals, and BBD had comparable percentages of classical, intermediate, and nonclassical monocytes. Expression of CD163, CD32, CD64, HLA-DR, CD38, CD40, CD86, CD91, CD11c, and CX3CR1 on monocytes did not differ between PLHIV and HIV-negative individuals, but it differed significantly from BBD. Principal component analysis revealed that 57.5% of PLHIV and 62.5% of HIV-negative individuals had a high monocyte activation profile compared with 2.9% of BBD. Cellular monocyte activation in the COBRA cohort was strongly associated with soluble markers of monocyte activation and inflammation in the CSF. Conclusions. People living with HIV and HIV-negative COBRA participants had high levels of cellular monocyte activation compared with age-matched BBD. High monocyte activation was predictive for inflammation in the CSF. PMID:28680905

  11. Environmental implications of high metal content in soils of a titanium mining zone in Kenya.

    Science.gov (United States)

    Maina, David M; Ndirangu, Douglas M; Mangala, Michael M; Boman, Johan; Shepherd, Keith; Gatari, Michael J

    2016-11-01

    Mining activities contribute to an increase of specific metal contaminants in soils. This may adversely affect plant life and consequently impact on animal and human health. The objective of this study was to obtain the background metal concentrations in soils around the titanium mining in Kwale County for monitoring its environmental impacts. Forty samples were obtained with half from topsoils and the other from subsoils. X-ray fluorescence spectrometry was used to determine the metal content of the soil samples. High concentrations of Ti, Mn, Fe, and Zr were observed where Ti concentrations ranged from 0.47 to 2.8 %; Mn 0.02 to 3.1 %; Fe 0.89 to 3.1 %; and Zr 0.05 to 0.85 %. Using ratios of elemental concentrations in topsoil to subsoil method and enrichment factors concept, the metals were observed to be of geogenic origin with no anthropogenic input. The high concentrations of Mn and Fe may increase their concentration levels in the surrounding agricultural lands through deposition, thereby causing contamination on the land and the cultivated food crops. The latter can cause adverse human health effects. In addition, titanium mining will produce tailings containing low-level titanium concentrations, which will require proper disposal to avoid increasing titanium concentrations in the soils of the region since it has been observed to be phytotoxic to plants at high concentrations. The results of this study will serve as reference while monitoring the environmental impact by the titanium mining activities.

  12. Assessment of Cr(VI-induced cytotoxicity and genotoxicity using high content analysis.

    Directory of Open Access Journals (Sweden)

    Chad M Thompson

    Full Text Available Oral exposure to high concentrations of hexavalent chromium [Cr(VI] induces intestinal redox changes, villus cytotoxicity, crypt hyperplasia, and intestinal tumors in mice. To assess the effects of Cr(VI in a cell model relevant to the intestine, undifferentiated (proliferating and differentiated (confluent Caco-2 cells were treated with Cr(VI, hydrogen peroxide or rotenone for 2-24 hours. DNA damage was then assessed by nuclear staining intensity of 8-hydroxydeoxyguanosine (8-OHdG and phosphorylated histone variant H2AX (γ-H2AX measured by high content analysis methods. In undifferentiated Caco-2, all three chemicals increased 8-OHdG and γ-H2AX staining at cytotoxic concentrations, whereas only 8-OHdG was elevated at non-cytotoxic concentrations at 24 hr. Differentiated Caco-2 were more resistant to cytotoxicity and DNA damage than undifferentiated cells, and there were no changes in apoptotic markers p53 or annexin-V. However, Cr(VI induced a dose-dependent translocation of the unfolded protein response transcription factor ATF6 into the nucleus. Micronucleus (MN formation was assessed in CHO-K1 and A549 cell lines. Cr(VI increased MN frequency in CHO-K1 only at highly cytotoxic concentrations. Relative to the positive control Mitomycin-C, Cr(VI only slightly increased MN frequency in A549 at mildly cytotoxic concentrations. The results demonstrate that Cr(VI genotoxicity correlates with cytotoxic concentrations, and that H2AX phosphorylation occurs at higher concentrations than oxidative DNA damage in proliferating Caco-2 cells. The findings suggest that in vitro genotoxicity of Cr(VI is primarily oxidative in nature at low concentrations. Implications for in vivo intestinal toxicity of Cr(VI will be discussed.

  13. Susceptibility Contrast in High Field MRI of Human Brain as a Function of Tissue Iron Content

    Science.gov (United States)

    Yao, Bing; Li, Tie-Qiang; van Gelderen, Peter; Shmueli, Karin; de Zwart, Jacco A.; Duyn, Jeff H.

    2009-01-01

    Magnetic susceptibility provides an important contrast mechanism for MRI. Increasingly, susceptibility-based contrast is being exploited to investigate brain tissue microstructure and to detect abnormal levels of brain iron as these have been implicated in a variety of neuro-degenerative diseases. However, it remains unclear to what extent magnetic susceptibility-related contrast at high field relates to actual brain iron concentrations. In this study, we performed susceptibility weighted imaging as a function of field strength on healthy brains in vivo and post-mortem brain tissues at 1.5T, 3T and 7T. Iron histology was performed on the tissue samples for comparison. The calculated susceptibility-related parameters R2* and signal frequency shift in four iron-rich regions (putamen, globus pallidus, caudate, and thalamus) showed an almost linear dependence (r=0.90 for R2*; r=0.83 for phase, p<0.01) on field strength, suggesting that potential ferritin saturation effects are not relevant to susceptibility-weighted contrast for field strengths up to 7T. The R2* dependence on the putative (literature-based) iron concentration was 0.048 Hz/Tesla/ppm. The histological data from brain samples confirmed the linear dependence of R2* on field strength and showed a slope against iron concentration of 0.0099 Hz/Tesla/ppm dry-weight, which is equivalent to 0.05 Hz/Tesla/ppm wet-weight and closely matched the calculated value in vivo. These results confirm the validity of using susceptibility-weighted contrast as an indicator of iron content in iron-rich brain regions. The absence of saturation effects opens the way to exploit the benefits of MRI at high field strengths for the detection of iron distributions with high sensitivity and resolution. PMID:19027861

  14. Effect of high contents of dietary animal-derived protein or carbohydrates on canine faecal microbiota

    Directory of Open Access Journals (Sweden)

    Hang Ingrid

    2012-06-01

    Full Text Available Abstract Background Considerable evidence suggests that food impacts both the gastro-intestinal (GI function and the microbial ecology of the canine GI tract. The aim of this study was to evaluate the influence of high-carbohydrate (HC, high-protein (HP and dry commercial (DC diets on the canine colonic microbiota in Beagle dogs. Diets were allocated according to the Graeco-Latin square design. For this purpose, microbial DNA was isolated from faecal samples and separated by density gradient centrifugation, resulting in specific profiling based on the guanine-cytosine content (%G + C. In addition, 16 S rRNA gene amplicons were obtained from the most abundant %G + C peaks and analysed by sequence analysis, producing a total of 720 non-redundant sequences (240 sequences per diet. Results The DC diet sample showed high abundance of representatives of the orders Clostridiales, Lactobacillales, Coriobacteriales and Bacteroidales. Sequence diversity was highest for DC diet samples and included representatives of the orders Lactobacillales and Bacteroidales, which were not detected in samples from the HP and HC diets. These latter two diets also had reduced levels of representatives of the family Lachnospiraceae, specifically Clostridial cluster XIVa. The HC diet favoured representatives of the order Erysipelotrichales, more specifically the Clostridial cluster XVIII, while the HP diet favoured representatives of the order Fusobacteriales. Conclusions This study detected Coriobacteriales in dog faeces, possibly due to the non-selective nature of the %G + C profiling method used in combination with sequencing. Moreover, our work demonstrates that the effect of diet on faecal microbiota can be explained based on the metabolic properties of the detected microbial taxa.

  15. Assessment of Cr(VI)-Induced Cytotoxicity and Genotoxicity Using High Content Analysis

    Science.gov (United States)

    Thompson, Chad M.; Fedorov, Yuriy; Brown, Daniel D.; Suh, Mina; Proctor, Deborah M.; Kuriakose, Liz; Haws, Laurie C.; Harris, Mark A.

    2012-01-01

    Oral exposure to high concentrations of hexavalent chromium [Cr(VI)] induces intestinal redox changes, villus cytotoxicity, crypt hyperplasia, and intestinal tumors in mice. To assess the effects of Cr(VI) in a cell model relevant to the intestine, undifferentiated (proliferating) and differentiated (confluent) Caco-2 cells were treated with Cr(VI), hydrogen peroxide or rotenone for 2–24 hours. DNA damage was then assessed by nuclear staining intensity of 8-hydroxydeoxyguanosine (8-OHdG) and phosphorylated histone variant H2AX (γ-H2AX) measured by high content analysis methods. In undifferentiated Caco-2, all three chemicals increased 8-OHdG and γ-H2AX staining at cytotoxic concentrations, whereas only 8-OHdG was elevated at non-cytotoxic concentrations at 24 hr. Differentiated Caco-2 were more resistant to cytotoxicity and DNA damage than undifferentiated cells, and there were no changes in apoptotic markers p53 or annexin-V. However, Cr(VI) induced a dose-dependent translocation of the unfolded protein response transcription factor ATF6 into the nucleus. Micronucleus (MN) formation was assessed in CHO-K1 and A549 cell lines. Cr(VI) increased MN frequency in CHO-K1 only at highly cytotoxic concentrations. Relative to the positive control Mitomycin-C, Cr(VI) only slightly increased MN frequency in A549 at mildly cytotoxic concentrations. The results demonstrate that Cr(VI) genotoxicity correlates with cytotoxic concentrations, and that H2AX phosphorylation occurs at higher concentrations than oxidative DNA damage in proliferating Caco-2 cells. The findings suggest that in vitro genotoxicity of Cr(VI) is primarily oxidative in nature at low concentrations. Implications for in vivo intestinal toxicity of Cr(VI) will be discussed. PMID:22905163

  16. Predicting in vivo anti-hepatofibrotic drug efficacy based on in vitro high-content analysis.

    Directory of Open Access Journals (Sweden)

    Baixue Zheng

    Full Text Available BACKGROUND/AIMS: Many anti-fibrotic drugs with high in vitro efficacies fail to produce significant effects in vivo. The aim of this work is to use a statistical approach to design a numerical predictor that correlates better with in vivo outcomes. METHODS: High-content analysis (HCA was performed with 49 drugs on hepatic stellate cells (HSCs LX-2 stained with 10 fibrotic markers. ~0.3 billion feature values from all cells in >150,000 images were quantified to reflect the drug effects. A systematic literature search on the in vivo effects of all 49 drugs on hepatofibrotic rats yields 28 papers with histological scores. The in vivo and in vitro datasets were used to compute a single efficacy predictor (E(predict. RESULTS: We used in vivo data from one context (CCl(4 rats with drug treatments to optimize the computation of E(predict. This optimized relationship was independently validated using in vivo data from two different contexts (treatment of DMN rats and prevention of CCl(4 induction. A linear in vitro-in vivo correlation was consistently observed in all the three contexts. We used E(predict values to cluster drugs according to efficacy; and found that high-efficacy drugs tended to target proliferation, apoptosis and contractility of HSCs. CONCLUSIONS: The E(predict statistic, based on a prioritized combination of in vitro features, provides a better correlation between in vitro and in vivo drug response than any of the traditional in vitro markers considered.

  17. The cytotoxicity of lead and uranium on rat osteoblastic cells is highly dependent on chemical speciation and cellular accumulation

    Energy Technology Data Exchange (ETDEWEB)

    Milgram, S.; Carriere, M.; Thiebault, C.; Gouget, B. [CEA Saclay, CNRS - UMR9956, Lab Pierre Sue, F-91198 Gif Sur Yvette, (France); Malval, L. [INSERM, E366, Lab Biol Tissue Osseux, St Etienne, (France)

    2007-07-01

    Complete text of publication follows: Uranium (U) and lead (Pb), as other heavy metals, present a strong chemical toxicity. After blood contamination, U and Pb, complexed with proteins or inorganic molecules are conveyed to target organs, the skeleton being the major long-term storage site. Once in bones, both metals are incorporated in the hydroxyapatite matrix by substitution with calcium. They can thus be released during re-modelling, which explains in part their toxicity. Although the clinical effects of these metals are well known, the cellular mechanisms of their action are not well understood. To investigate the biological effects of U and Pb acute exposure on osteoblasts, ROS17/2.8 cells were exposed to Pb or U [0-1 mM] for 24 h. The most relevant chemical and physical states, namely the most likely forms (species) of the toxics in contact with cells after blood contamination were selected for cell exposure. For each metal species, Pb and U toxicity were assessed through cell viability assay. The results show that whatever the speciation, U chemical toxicity to bone cells is far lower than Pb toxicity. Pb appears to be cytotoxic when left free in the exposure medium or when it is complexed with bicarbonate, cysteine or citrate, but not with albumin or phosphate (an insoluble form of Pb). In order to explain these differences in sensitivity between different metals and metal chemical species, time-course and dose-response curves of cellular accumulation at lethal or sub-lethal doses were drawn by direct elemental analysis of metal concentrations in digested cell pellets, using Inductive Coupling Plasma Mass Spectroscopy. These showed a clear correlation between toxicity and cellular accumulation. Also, Pb induces an inhibition of ALP activity after 24 h exposure to sub-lethal doses, which is speciation-dependent and again correlates with cellular accumulation. Phenotypic effects of U are under investigation. In addition, electron-microscopic observation of

  18. Toward high-content screening of mitochondrial morphology and membrane potential in living cells

    NARCIS (Netherlands)

    Iannetti, E.F.; Willems, P.H.G.M.; Pellegrini, M.; Beyrath, J.D.; Smeitink, J.; Blanchet, L.M.; Koopman, W.J.H.

    2015-01-01

    Mitochondria are double membrane organelles involved in various key cellular processes. Governed by dedicated protein machinery, mitochondria move and continuously fuse and divide. These "mitochondrial dynamics" are bi-directionally linked to mitochondrial and cell functional state in space and time

  19. High β phase content in PVDF/CoFe2O4 nanocomposites induced by DC magnetic fields

    Science.gov (United States)

    Jiang, Shenglin; Wan, Hongyan; Liu, Huan; Zeng, Yike; Liu, Jianguo; Wu, Yunyi; Zhang, Guangzu

    2016-09-01

    Flexible ferroelectrics being exploited as energy harvesting and conversion materials are highly desirable for wearable and skin-mountable electronic devices. As one of the most typical ferroelectric polymers, poly(vinylidene fluoride) (PVDF) has been widely used in modern electronic systems and devices, whose ferroelectric performance relies heavily on its β phase content. In this work, to achieve high-β-phase-content PVDF, we first introduced CoFe2O4 nanoparticles into PVDF. With the incorporation of CoFe2O4 nanoparticles used as an effective polymer nucleation agent, the percentage of the β phase in the PVDF has been significantly enhanced, e.g., 84% in the nanocomposite with 5 wt. % CoFe2O4 versus only 73% in the pure PVDF. In order to further increase the β phase content in PVDF, we subsequently proposed an easily realized strategy. By applying DC magnetic fields during the solution-casting process of the PVDF/CoFe2O4 nanocomposites, a further improved β phase content as high as 95% can be achieved. The further improvement of the β phase content is attributable to the tensile stress at the CoFe2O4/PVDF interfaces created by the coupling of magnetic field and CoFe2O4 by means of the magnetostriction effect. The high β-phase content makes the PVDF/CoFe2O4 nanocomposites a promising candidate for flexible and wearable electronic device applications.

  20. Robo-Lector – a novel platform for automated high-throughput cultivations in microtiter plates with high information content

    Directory of Open Access Journals (Sweden)

    Kensy Frank

    2009-08-01

    culture plate, where subsequently similar growth kinetics could be obtained. Conclusion The Robo-Lector generates extensive kinetic data in high-throughput cultivations, particularly for biomass and fluorescence protein formation. Based on the non-invasive on-line-monitoring signals, actions of the liquid-handling robot can easily be triggered. This interaction between the robot and the BioLector (Robo-Lector combines high-content data generation with systematic high-throughput experimentation in an automated fashion, offering new possibilities to study biological production systems. The presented platform uses a standard liquid-handling workstation with widespread automation possibilities. Thus, high-throughput cultivations can now be combined with small-scale downstream processing techniques and analytical assays. Ultimately, this novel versatile platform can accelerate and intensify research and development in the field of systems biology as well as modelling and bioprocess optimization.

  1. Biodrying process: A sustainable technology for treatment of municipal solid waste with high moisture content.

    Science.gov (United States)

    Tom, Asha P; Pawels, Renu; Haridas, Ajit

    2016-03-01

    Municipal solid waste with high moisture content is the major hindrance in the field of waste to energy conversion technologies and here comes the importance of biodrying process. Biodrying is a convective evaporation process, which utilizes the biological heat developed from the aerobic reactions of organic components. The numerous end use possibilities of the output are making the biodrying process versatile, which is possible by achieving the required moisture reduction, volume reduction and bulk density enhancement through the effective utilization of biological heat. In the present case study the detailed research and development of an innovative biodrying reactor has been carried out for the treatment of mixed municipal solid waste with high moisture content. A pilot scale biodrying reactor of capacity 565 cm(3) was designed and set up in the laboratory. The reactor dimensions consisted of an acrylic chamber of 60 cm diameter and 200 cm height, and it was enveloped by an insulation chamber. The insulation chamber was provided to minimise the heat losses through the side walls of the reactor. It simulates the actual condition in scaling up of the reactor, since in bigger scale reactors the heat losses through side walls will be negligible while comparing the volume to surface area ratio. The mixed municipal solid waste with initial moisture content of 61.25% was synthetically prepared in the laboratory and the reactor was fed with 109 kg of this substrate. Aerobic conditions were ensured inside the reactor chamber by providing the air at a constant rate of 40 litre per minute, and the direction of air flow was from the specially designed bottom air chamber to the reactor matrix top. The self heating inside reactor matrix was assumed in the range of 50-60°C during the design stage. Innovative biodrying reactor was found to be efficiently working with the temperature inside the reactor matrix rising to a peak value of 59°C by the fourth day of experiment (the

  2. Submerged Cultivation of Mycelium with High Ergothioneine Content from the Culinary-Medicinal Golden Oyster Mushroom, Pleurotus citrinopileatus (Higher Basidiomycetes).

    Science.gov (United States)

    Lin, Shin-Yi; Chien, Shih-Chang; Wang, Sheng-Yang; Mau, Jeng-Leun

    2015-01-01

    The optimization of submerged culture of the culinary-medicinal golden oyster mushroom, Pleurotus citrinopileatus, was studied using a one-factor-at-a-time, two-stage stimulation and central composite rotatable design to produce mycelia with high ergothioneine content. The optimal culture conditions for mycelia harvested at day 22 were a temperature of 25°C, an inoculation ratio of 5%, 2% glucose, 0.5% yeast extract, and adjustment of the initial pH value to 10. The biomass and ergothioneine content were 8.28 g/L and 10.65 mg/g dry weight (dw), respectively. The addition of an amino acid precursor increased the ergothioneine content of mycelia; cysteine was the most effective. In addition, the results obtained from central composite rotatable design showed that the recommended combination for cysteine, histidine, and methionine was 8, 4, and 0.5 mmol/L, respectively. The predicted ergothioneine content was 13.90 mg/g dw, whereas the experimental maximal ergothioneine content was 14.57 mg/g dw. With the addition of complex precursors and under optimal culture conditions, mycelia harvested at days 16-20 had higher ergothioneine content. Accordingly, the information obtained could be used to produce mycelia with high ergothioneine content.

  3. Analysis of SSLP and Soluble Protein Contents in Leaves of Mutants Induced by High Pressure in Rice (Oryza sativa)

    Institute of Scientific and Technical Information of China (English)

    HE Xiu-ying; XU Shi-ping; LIAO Yao-ping; MAO Xing-xue; WENG Ke-nan; CHEN Zhao-ming; CHEN Yue-han; XIAO Wan-sheng

    2004-01-01

    Rice variety Yuexiangzhan and its mutants induced by high pressure were studied using microsatellite markers and soluble protein content analyses. Eleven of the 88 microsatellite primer pairs showed evident polymorphisms repeatedly, and the polymorphic frequencies were 3.4-11.3% between the mutants and Yuexiangzhan. The polymorphic markers were randomly located on chromosomes. The more similar the plant types of the mutants like their original variety, the less polymorphic loci were detected. In addition, there was variation in the soluble protein contents among the leaves of mutants,and the contents were significantly lower than those of the original variety.

  4. Highly recommended!’ The content characteristics and perceived usefulness of online consumer reviews

    NARCIS (Netherlands)

    L.M. Willemsen; P.C. Neijens; F. Bronner; J.A. de Ridder

    2011-01-01

    The aim of the present study was to gain a better understanding of the content characteristics that make online consumer reviews a useful source of consumer information. To this end, we content analyzed reviews of experience and search products posted on Amazon.com (N = 400). The insights derived fr

  5. Initial Characterization of Colombian High School Physics Teachers' Pedagogical Content Knowledge on Electric Fields

    Science.gov (United States)

    Melo-Niño, Lina Viviana; Cañada, Florentina; Mellado, Vicente

    2017-01-01

    We explore the initial characterization of the pedagogical content knowledge of four, in-service, Colombian pre-university secondary education physics teachers on the concept of electric field. Two of them teach the content in English as a second language. The aim of the study was to obtain an image of the participants' teaching of electric field…

  6. Initial Characterization of Colombian High School Physics Teachers' Pedagogical Content Knowledge on Electric Fields

    Science.gov (United States)

    Melo-Niño, Lina Viviana; Cañada, Florentina; Mellado, Vicente

    2017-01-01

    We explore the initial characterization of the pedagogical content knowledge of four, in-service, Colombian pre-university secondary education physics teachers on the concept of electric field. Two of them teach the content in English as a second language. The aim of the study was to obtain an image of the participants' teaching of electric field…

  7. Increased BK viremia and progression to BK-virus nephropathy following high-dose intravenous immunoglobulin for acute cellular rejection.

    Science.gov (United States)

    Boonyapredee, Maytee; Knight, Kendral; Little, Dustin

    2014-06-01

    BK virus nephropathy and cellular rejection are common causes of allograft dysfunction in renal transplant recipients. The two can be difficult to distinguish on allograft biopsy and can be present simultaneously. Management of the patient with coexistent BK infection and rejection is complicated by the conflicting ideals of decreasing immunosuppression to treat the former and increasing immunosuppression to treat the latter. The authors present the case of a 57-year-old renal transplant recipient who underwent allograft biopsy 8 weeks post-transplant for evaluation of increased serum creatinine in the setting of BK viremia (BKV). Biopsy revealed Banff classification 1b acute cellular rejection, with insufficient evidence to diagnose BK virus-associated nephropathy. The patient was administered intravenous immune globulin (IVIG), with no other changes in immunosuppressive therapy. Plasma and urine BK increased exponentially following IVIG administration, and allograft function further deteriorated. Repeat biopsy showed overt BK viral nephropathy, and BKV and creatinine decreased only after reduction in immunosuppression and initiation of leflunomide. Although case series have suggested a potential role for IVIG in the setting of BK infection, further study is needed to define the safety and efficacy of this approach. Reprint & Copyright © 2014 Association of Military Surgeons of the U.S.

  8. Study of elastic-plastic damage model of cement consolidated soil with high organic content

    Institute of Scientific and Technical Information of China (English)

    CHEN Huie; WANG Qing; CAI Keyi

    2009-01-01

    On the basis of elastic-plastic damage model of cement consolidated soil, the authors took organic contents into reasonable damage variable evolution equation in order to seek relation between the organic contents and parameters in the equation, and established the elastic-plastic damage model of cement consolidated soil considering organic contents. The results show that the parameters change correspondingly with difference of the organic contents. The higher the organic contents are, the less the valves of the parameters such as elastic modulus (E), material parameters (K, n) and damage evolution parameter (ε) become, but the larger strain damage threshold value (εd) of the sample is. Meanwhile, the calculation results obtained from established model are compared with the test data in the condition of common indoors test, which is testified with reliability.

  9. On forecasting ionospheric total electron content responses to high-speed solar wind streams

    Directory of Open Access Journals (Sweden)

    Meng Xing

    2016-01-01

    Full Text Available Conditions in the ionosphere have become increasingly important to forecast, since more and more spaceborne and ground-based technological systems rely on ionospheric weather. Here we explore the feasibility of ionospheric forecasts with the current generation of physics-based models. In particular, we focus on total electron content (TEC predictions using the Global Ionosphere-Thermosphere Model (GITM. Simulations are configured in a forecast mode and performed for four typical high-speed-stream events during 2007–2012. The simulated TECs are quantified through a metric, which divides the globe into a number of local regions and robustly differentiates between quiet and disturbed periods. Proposed forecast products are hourly global maps color-coded by the TEC disturbance level of each local region. To assess the forecasts, we compare the simulated TEC disturbances with global TEC maps derived from Global Positioning System (GPS satellite observations. The forecast performance is found to be merely acceptable, with a large number of regions where the observed variations are not captured by the simulations. Examples of model-data agreements and disagreements are investigated in detail, aiming to understand the model behavior and improve future forecasts. For one event, we identify two adjacent regions with similar TEC observations but significant differences in how local chemistry versus plasma transport contribute to electron density changes in the simulation. Suggestions for further analysis are described.

  10. Belite cement clinker from coal fly ash of high Ca content. Optimization of synthesis parameters

    Energy Technology Data Exchange (ETDEWEB)

    A. Guerrero; S. Goni; I. Campillo; A. Moragues [Instituto de Ciencias de la Construccion ' Eduardo Torroja' (CSIC), Madrid (Spain)

    2004-06-01

    The optimization of parameters of synthesis of belite cement clinker from Spanish coal fly ash of high Ca content is presented in this paper. The investigations were prompted by a need to reduce the CO{sub 2} emissions produced by the cement industry. Belite Ca{sub 2}SiO{sub 4} synthesis produces less CO{sub 2} than alite, the main component of Portland cement clinker. The synthesis process is based on the hydrothermal-calcination-route of the fly ash without extra additions. The hydrothermal treatment was carried out in demineralized water and a 1 M NaOH solution for 4 h at the temperatures of 100, 150, and 200{sup o} C. The precursors obtained during the hydrothermal treatment were heated at temperatures of 700, 800, 900, and 1000{sup o}C. The changes of fly ash composition after the different treatments were characterized by X-ray diffraction (XRD), FT infrared (FTIR) spectroscopy, surface area (BET-N{sub 2}), and thermal analyses. From the results obtained it was concluded that the optimum temperature of the hydrothermal treatment was 200{sup o}C, and the optimum temperature for obtaining the belite cement clinker was 800{sup o}C. 12 refs., 11 figs., 2 tabs.

  11. Recirculation of Chilean copper smelting dust with high impurities contents to the smelting process

    Energy Technology Data Exchange (ETDEWEB)

    Sano, H.; Fujisawa, T. [Nagoya Univ., Nagoya (Japan). EcoTopia Science Inst.; Montenegro, V. [Nagoya Univ., Nagoya (Japan). Dept. of Materials Science and Engineering

    2007-07-01

    Dust generated during the copper smelting process is generally stabilized using hydrometallurgical methods as it contains high concentrations of arsenic. In this laboratory study, dust was recirculated during the smelting process in order to recover more copper and decrease dust emissions while recovering more copper. The behaviour of impurities and their influence on matte quality was also investigated. Industrial matte, flue dust, slag, and copper concentrates from a Chilean smelter were used as test materials. Dust recirculation tests were conducted in a simulated electric furnace. Off-gases were collected in a reaction tube, and the condensed volatile matter, slag, and matte phases were analyzed for their elemental content by inductively coupled plasma atomic emission spectrometry. The distribution of arsenic (As); antimony (Sb), bismuth (Bi), lead (Pb), and zinc (Zn) were investigated by varying the amounts of dust recirculating to the smelting stage with 21 per cent of the oxygen. Results showed that distributions of all analyzed elements increased with recirculation. It was concluded that copper can be recovered using the dust recirculation technique. However, impurities may limit the efficacy of the dust recirculation process. 6 refs., 3 tabs., 4 figs.

  12. On forecasting ionospheric total electron content responses to high-speed solar wind streams

    Science.gov (United States)

    Meng, Xing; Mannucci, Anthony J.; Verkhoglyadova, Olga P.; Tsurutani, Bruce T.

    2016-04-01

    Conditions in the ionosphere have become increasingly important to forecast, since more and more spaceborne and ground-based technological systems rely on ionospheric weather. Here we explore the feasibility of ionospheric forecasts with the current generation of physics-based models. In particular, we focus on total electron content (TEC) predictions using the Global Ionosphere-Thermosphere Model (GITM). Simulations are configured in a forecast mode and performed for four typical high-speed-stream events during 2007-2012. The simulated TECs are quantified through a metric, which divides the globe into a number of local regions and robustly differentiates between quiet and disturbed periods. Proposed forecast products are hourly global maps color-coded by the TEC disturbance level of each local region. To assess the forecasts, we compare the simulated TEC disturbances with global TEC maps derived from Global Positioning System (GPS) satellite observations. The forecast performance is found to be merely acceptable, with a large number of regions where the observed variations are not captured by the simulations. Examples of model-data agreements and disagreements are investigated in detail, aiming to understand the model behavior and improve future forecasts. For one event, we identify two adjacent regions with similar TEC observations but significant differences in how local chemistry versus plasma transport contribute to electron density changes in the simulation. Suggestions for further analysis are described.

  13. Synthesis and high content cell-based profiling of simplified analogues of the microtubule stabilizer (+)-discodermolide.

    Science.gov (United States)

    Minguez, Jose M; Giuliano, Kenneth A; Balachandran, Raghavan; Madiraju, Charitha; Curran, Dennis P; Day, Billy W

    2002-12-01

    (+)-Discodermolide, a C24:4, trihydroxylated, octamethyl, carbamate-bearing fatty acid lactone originally isolated from a Caribbean sponge, has proven to be the most potent of the microtubule-stabilizing agents. Recent studies suggest that it or its analogues may have advantages over other classes of microtubule-stabilizing agents. (+)-Discodermolide's complex molecular architecture has made structure-activity relationship analysis in this class of compounds a formidable task. The goal of this study was to prepare simplified analogues of (+)-discodermolide and to analyze their biological activities to expand structure-activity relationships. A small library of analogues was prepared wherein the (+)-discodermolide methyl groups at C-14 and C-16 and the C-7 hydroxyl were removed, and the lactone was replaced by simple esters. The library components were analyzed for microtubule-stabilizing actions in vitro, antiproliferative activity against a small panel of human carcinoma cells, and cell signaling, microtubule architecture and mitotic spindle alterations by a multiparameter fluorescence cell-based screening technique. The results show that even drastic structural simplification can lead to analogues with actions related to microtubule targeting and signal transduction, but that these subtle effects were illuminated only through the high information content cell-based screen.

  14. A Multivariate Computational Method to Analyze High-Content RNAi Screening Data.

    Science.gov (United States)

    Rameseder, Jonathan; Krismer, Konstantin; Dayma, Yogesh; Ehrenberger, Tobias; Hwang, Mun Kyung; Airoldi, Edoardo M; Floyd, Scott R; Yaffe, Michael B

    2015-09-01

    High-content screening (HCS) using RNA interference (RNAi) in combination with automated microscopy is a powerful investigative tool to explore complex biological processes. However, despite the plethora of data generated from these screens, little progress has been made in analyzing HC data using multivariate methods that exploit the full richness of multidimensional data. We developed a novel multivariate method for HCS, multivariate robust analysis method (M-RAM), integrating image feature selection with ranking of perturbations for hit identification, and applied this method to an HC RNAi screen to discover novel components of the DNA damage response in an osteosarcoma cell line. M-RAM automatically selects the most informative phenotypic readouts and time points to facilitate the more efficient design of follow-up experiments and enhance biological understanding. Our method outperforms univariate hit identification and identifies relevant genes that these approaches would have missed. We found that statistical cell-to-cell variation in phenotypic responses is an important predictor of hits in RNAi-directed image-based screens. Genes that we identified as modulators of DNA damage signaling in U2OS cells include B-Raf, a cancer driver gene in multiple tumor types, whose role in DNA damage signaling we confirm experimentally, and multiple subunits of protein kinase A. © 2015 Society for Laboratory Automation and Screening.

  15. Determination of purine contents in different parts of pork and beef by high performance liquid chromatography.

    Science.gov (United States)

    Rong, Shengzhong; Zou, Lina; Zhang, Yannan; Zhang, Guangteng; Li, Xiaoxia; Li, Miaojing; Yang, Fenghua; Li, Chunmei; He, Yingjuan; Guan, Hongjun; Guo, Yupeng; Wang, Dong; Cui, Xinyu; Ye, Hongting; Liu, Fenghai; Pan, Hongzhi; Yang, Yuexin

    2015-03-01

    Determination of adenine, hypoxanthine, guanine and xanthine in different parts of pork and beef using high performance liquid chromatography was described. Chromatographic separation was carried out on Waters Atlantis T3 column (4.6 mm × 250 mm × 5 μm) with column temperature at 30 °C. The mobile phase contained 99% 10.0 mmol/L ammonium formate solution at pH 3.6 and 1.0% methanol. Chromatography was achieved at a flow rate of 1.0 mL/min and detection wavelength at 254 nm. The results indicated that total purine amounts in pork rump and beef sirloin were higher than those in other parts (Ppurine bases were hypoxanthine and adenine, and hypoxanthine content was the most highest in all samples (Ppurine and uricogenic purine base, we suggest that excess consumption of them be avoid, whereas pork loin chop and beef rib eye are more suitable for a low-purine diet. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. A high-content platform to characterise human induced pluripotent stem cell lines

    Science.gov (United States)

    Leha, Andreas; Moens, Nathalie; Meleckyte, Ruta; Culley, Oliver J.; Gervasio, Mia K.; Kerz, Maximilian; Reimer, Andreas; Cain, Stuart A.; Streeter, Ian; Folarin, Amos; Stegle, Oliver; Kielty, Cay M.; Durbin, Richard; Watt, Fiona M.; Danovi, Davide

    2016-01-01

    Induced pluripotent stem cells (iPSCs) provide invaluable opportunities for future cell therapies as well as for studying human development, modelling diseases and discovering therapeutics. In order to realise the potential of iPSCs, it is crucial to comprehensively characterise cells generated from large cohorts of healthy and diseased individuals. The human iPSC initiative (HipSci) is assessing a large panel of cell lines to define cell phenotypes, dissect inter- and intra-line and donor variability and identify its key determinant components. Here we report the establishment of a high-content platform for phenotypic analysis of human iPSC lines. In the described assay, cells are dissociated and seeded as single cells onto 96-well plates coated with fibronectin at three different concentrations. This method allows assessment of cell number, proliferation, morphology and intercellular adhesion. Altogether, our strategy delivers robust quantification of phenotypic diversity within complex cell populations facilitating future identification of the genetic, biological and technical determinants of variance. Approaches such as the one described can be used to benchmark iPSCs from multiple donors and create novel platforms that can readily be tailored for disease modelling and drug discovery. PMID:26608109

  17. Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy.

    Science.gov (United States)

    Görlitz, Frederik; Kelly, Douglas J; Warren, Sean C; Alibhai, Dominic; West, Lucien; Kumar, Sunil; Alexandrov, Yuriy; Munro, Ian; Garcia, Edwin; McGinty, James; Talbot, Clifford; Serwa, Remigiusz A; Thinon, Emmanuelle; da Paola, Vincenzo; Murray, Edward J; Stuhmeier, Frank; Neil, Mark A A; Tate, Edward W; Dunsby, Christopher; French, Paul M W

    2017-01-18

    We present an open source high content analysis instrument utilizing automated fluorescence lifetime imaging (FLIM) for assaying protein interactions using Förster resonance energy transfer (FRET) based readouts of fixed or live cells in multiwell plates. This provides a means to screen for cell signaling processes read out using intramolecular FRET biosensors or intermolecular FRET of protein interactions such as oligomerization or heterodimerization, which can be used to identify binding partners. We describe here the functionality of this automated multiwell plate FLIM instrumentation and present exemplar data from our studies of HIV Gag protein oligomerization and a time course of a FRET biosensor in live cells. A detailed description of the practical implementation is then provided with reference to a list of hardware components and a description of the open source data acquisition software written in µManager. The application of FLIMfit, an open source MATLAB-based client for the OMERO platform, to analyze arrays of multiwell plate FLIM data is also presented. The protocols for imaging fixed and live cells are outlined and a demonstration of an automated multiwell plate FLIM experiment using cells expressing fluorescent protein-based FRET constructs is presented. This is complemented by a walk-through of the data analysis for this specific FLIM FRET data set.

  18. Robust content-dependent high-fidelity watermark for tracking in digital cinema

    Science.gov (United States)

    Lubin, Jeffrey; Bloom, Jeffrey A.; Cheng, Hui

    2003-06-01

    Forensic digital watermarking is a promising tool in the fight against piracy of copyrighted motion imagery content, but to be effective it must be (1) imperceptibly embedded in high-definition motion picture source, (2) reliably retrieved, even from degraded copies as might result from camcorder capture and subsequent very-low-bitrate compression and distribution on the Internet, and (3) secure against unauthorized removal. No existing watermarking technology has yet to meet these three simultaneous requirements of fidelity, robustness, and security. We describe here a forensic watermarking approach that meets all three requirements. It is based on the inherent robustness and imperceptibility of very low spatiotemporal frequency watermark carriers, and on a watermark placement technique that renders jamming attacks too costly in picture quality, even if the attacker has complete knowledge of the embedding algorithm. The algorithm has been tested on HD Cinemascope source material exhibited in a digital cinema viewing room. The watermark is imperceptible, yet recoverable after exhibition capture with camcorders, and after the introduction of other distortions such as low-pass filtering, noise addition, geometric shifts, and the manipulation of brightness and contrast.

  19. In vitro oxidation of high polydimethylsiloxane content biomedical polyurethanes: correlation with the microstructure.

    Science.gov (United States)

    Hernandez, Rebeca; Weksler, Jadwiga; Padsalgikar, Ajay; Runt, James

    2008-11-01

    The resistance to in vitro metal ion oxidation of a polydimethylsiloxane (PDMS)-containing thermoplastic polyurethane elastomer (Elast-Eon) is compared with that of a polyurethane consisting of the same hard segment chemistry and content, but with aliphatic polycarbonate soft segments (PCU). Scanning electron microscopy and attenuated total reflectance Fourier transform infrared spectroscopy were used to assess changes in surface morphology and chemistry. The extent of bulk degradation was assessed indirectly by dynamic mechanical analysis and small-angle X-ray scattering experiments. The findings indicate that Elast-Eon is more resistant to oxidation than the PCU, because of the presence of the PDMS soft segments as well as its phase separated microstructure. The PCU exhibits a rather high degree of intermixing between hard and soft segments, rendering the hard segments dissolved or trapped in the soft phase more susceptible to oxidative conditions. By contrast, we propose that the existence of a completely phase separated PDMS soft phase in Elast-Eon protects the remainder of the segments from oxidation.

  20. Nonionic Polymerizable Emulsifier in High-Solids-Content Acrylate Emulsion Polymerization

    Institute of Scientific and Technical Information of China (English)

    LU Deping; HUANG Hongzhi; SHEN Ling; XIE Jin; GUAN Rong

    2012-01-01

    Stable high-solids-content acrylate emulsion were obtained with a nonionic polymerizable emulsifier allyloxy nonylphenoxy poly (ethyleneoxy) (10) ether (ANPEO10),and a conventional emulsifier OP-10 as a reference sample.1H NMR proves that the polymerizable emulsifier ANPEO10 has been incorporated into the resulted acrylate polymers.TEM demonstrates that there are some differences in the particle morphologies.AFM proves that the polymerizable emulsifier ANPEO10 migrating to the surface of the emulsion film was much less than the conventional emulsifier OP-10.The polymerizable emulsifier ANPEO10 can enhance the adhesion with glass plate compared to the conventional emulsifier.Furthermore,with increasing amount of emulsifier,the surface free energy of the films first decreased and then increased,and the adhesion with glass plate is initially enhanced and then attenuated.The water-resistance and solvent-resistance of the films prepared by the polymerizable emulsifier ANPEO10 are superior to those prepared by the conventional emulsifier OP-10.

  1. STUDIES ON HIGH SOLID CONTENT AND STABLE EMULSIFIER—FREE EMULSION POLYMERIZATION

    Institute of Scientific and Technical Information of China (English)

    TangGuangliang; HaoGuangjie; 等

    1998-01-01

    In this article,AHPS(3-allyloxy-2-hydroxy-1-propanesulfonate salt) was synthesized to use in emulsifier-free emulsion polymerization of BA/BMA/MMA system for high solid content latexes.Storage stability,flow behavior of the latexes,morphology of the obtained latex particles,dynamic viscoelastic behavior,tensile strength and water resistance properties of the resulted copolymers were investigated.The experimental results show that with the addition of AHPS,stability of the emulsion is greatly improved that there appears no apparent precipitation during the polymerization and storage at room temperature for 6 months and at -10℃ for four months.Flow of the latexes follows the Bingham body laws,diameter of the latex particles is about 0.6μm,of which is larger than that of by conventional emulsion polymerization(0.12μm).In addition,not only water resistance of the copolymers obtained by emulsifier-free emulstion polymerization is greatly improved,but also tensile strength is obviously enhanced.

  2. Content-based high-resolution remote sensing image retrieval with local binary patterns

    Science.gov (United States)

    Wang, A. P.; Wang, S. G.

    2006-10-01

    Texture is a very important feature in image analysis including content-based image retrieval (CBIR). A common way of retrieving images is to calculate the similarity of features between a sample images and the other images in a database. This paper applies a novel texture analysis approach, local binary patterns (LBP) operator, to 1m Ikonos images retrieval and presents an improved LBP histogram spatially enhanced LBP (SEL) histogram with spatial information by dividing the LBP labeled images into k*k regions. First different neighborhood P and scale factor R were chosen to scan over the whole images, so that their labeled LBP and local variance (VAR) images were calculated, from which we got the LBP, LBP/VAR, and VAR histograms and SEL histograms. The histograms were used as the features for CBIR and a non-parametric statistical test G-statistic was used for similarity measure. The result showed that LBP/VAR based features got a very high retrieval rate with certain values of P and R, and SEL features that are more robust to illumination changes than LBP/VAR also obtained higher retrieval rate than LBP histograms. The comparison to Gabor filter confirmed the effectiveness of the presented approach in CBIR.

  3. Pulse laser assisted MOVPE for InGaN with high indium content

    Energy Technology Data Exchange (ETDEWEB)

    Kawaguchi, Norihito [Technical Development and Engineering Center, Ishikawajima-Harima Heavy Industries Co., Ltd., Yokohama 235-8501 (Japan); Dept. of Applied Chemistry, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan); Hida, Kennosuke; Kangawa, Yoshihiro; Kumagai, Yoshinao; Koukitu, Akinori [Dept. of Applied Chemistry, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588 (Japan)

    2004-09-01

    In{sub 0.53}Ga{sub 0.47}N film was grown at 600 C by Nd:YAG pulse laser assisted MOVPE. The optical and structural properties of the film were compared with that grown without laser assistance at the same condition. The results of XRD measurements showed that the crystallinity of the film grown with laser was better than that of the one grown without laser. The surface morphology and cross-sectional SEM image of the film grown with laser revealed that there were no In droplets on the film. The band-edge emission of the film grown with laser at room temperature and 77 K was observed at 840 nm. The results of micro-Raman measurement showed that the film grown with laser had better crystalline structure than that of the film grown without laser and the radiative recombination which contributed to photoluminescence mainly occurred at In{sub 0.53}Ga{sub 0.47}N region. Those results imply that pulse laser enhances the surface migration and reaction of elements in spite of low-growth temperature. We suggest that pulse laser assisted technique is effective for low-temperature growth of InGaN with high indium content. (copyright 2004 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  4. Pulse laser assisted MOVPE for InGaN with high indium content

    Science.gov (United States)

    Kawaguchi, Norihito; Hida, Ken-Nosuke; Kangawa, Yoshihiro; Kumagai, Yoshinao; Koukitu, Akinori

    2004-09-01

    In0.53Ga0.47N film was grown at 600 °C by Nd:YAG pulse laser assisted MOVPE. The optical and structural properties of the film were compared with that grown without laser assistance at the same condition. The results of XRD measurements showed that the crystallinity of the film grown with laser was better than that of the one grown without laser. The surface morphology and cross-sectional SEM image of the film grown with laser revealed that there were no In droplets on the film. The band-edge emission of the film grown with laser at room temperature and 77 K was observed at 840 nm. The results of micro-Raman measurement showed that the film grown with laser had better crystalline structure than that of the film grown without laser and the radiative recombination which contributed to photoluminescence mainly occurred at In0.53Ga0.47N region. Those results imply that pulse laser enhances the surface migration and reaction of elements in spite of low-growth temperature. We suggest that pulse laser assisted technique is effective for low-temperature growth of InGaN with high indium content.

  5. Discovering Molecules That Regulate Efferocytosis Using Primary Human Macrophages and High Content Imaging.

    Directory of Open Access Journals (Sweden)

    Sandra Santulli-Marotto

    Full Text Available Defective clearance of apoptotic cells can result in sustained inflammation and subsequent autoimmunity. Macrophages, the "professional phagocyte" of the body, are responsible for efficient, non-phlogistic, apoptotic cell clearance. Controlling phagocytosis of apoptotic cells by macrophages is an attractive therapeutic opportunity to ameliorate inflammation. Using high content imaging, we have developed a system for evaluating the effects of antibody treatment on apoptotic cell uptake in primary human macrophages by comparing the Phagocytic Index (PI for each antibody. Herein we demonstrate the feasibility of evaluating a panel of antibodies of unknown specificities obtained by immunization of mice with primary human macrophages and show that they can be distinguished based on individual PI measurements. In this study ~50% of antibodies obtained enhance phagocytosis of apoptotic cells while approximately 5% of the antibodies in the panel exhibit some inhibition. Though the specificities of the majority of antibodies are unknown, two of the antibodies that improved apoptotic cell uptake recognize recombinant MerTK; a receptor known to function in this capacity in vivo. The agonistic impact of these antibodies on efferocytosis could be demonstrated without addition of either of the MerTK ligands, Gas6 or ProS. These results validate applying the mechanism of this fundamental biological process as a means for identification of modulators that could potentially serve as therapeutics. This strategy for interrogating macrophages to discover molecules regulating apoptotic cell uptake is not limited by access to purified protein thereby increasing the possibility of finding novel apoptotic cell uptake pathways.

  6. A high-content analysis toolbox permits dissection of diverse signaling pathways for T lymphocyte polarization.

    Science.gov (United States)

    Freeley, Michael; Bakos, Gabor; Davies, Anthony; Kelleher, Dermot; Long, Aideen; Dunican, Dara J

    2010-06-01

    RNA interfering (RNAi) screening strategies offer the potential to elucidate the signaling pathways that regulate integrin and adhesion receptor-mediated changes in T lymphocyte morphology. Of crucial importance, however, is the definition of key sets of parameters that will provide accurate, quantitative, and nonredundant information to flag relevant hits in such assays. In this study, the authors have used an image-based high-content analysis (HCA) technology platform and a panel of 24 pharmacological inhibitors, at a range of concentrations, to define key sets of parameters that enables sensitive and quantitative effects on integrin (LFA-1)-mediated lymphocyte morphology to be evaluated. In particular, multiparametric analysis of lymphocyte morphology that was based on intracellular staining of both the F-actin and alpha-tubulin cytoskeleton resulted in improved ability to discriminate morphological behavior compared to F-actin staining alone. Morphological and fluorescence intensity/distribution profiling of pharmacologically treated lymphocytes stimulated with integrin (LFA-1) and adhesion receptors (CD44) also revealed notable differences in their sensitivity to inhibitors. The assay described here may be used in HCA strategies such as RNAi screening assays to elucidate the signaling pathways and molecules that regulate integrin/adhesion receptor-mediated T lymphocyte polarization.

  7. Developing E-Learning Based on Animation Content for Improving Mathematical Connection Abilities in High School Students

    OpenAIRE

    Dedi Rohendi

    2012-01-01

    The purpose of this paper is to develop e-learning based on animation content for improving mathematical connection abilities in senior high school students. The e-learning was developed by using Moddle and the animation content was developed by using macromedia flash. To get the student mathematical conection abilities it uses the instruments of mathematical tests before and after teaching and learning process. The data were analyzed by using t-test and gain value test. The study found that ...

  8. Effect of High Temperature on Sucrose Content and Sucrose Cleaving Enzyme Activity in Rice Grain During the Filling Stage

    Institute of Scientific and Technical Information of China (English)

    LI Tian; LIU Qi-hua; Ryu OHSUGI; Tohru YAMAGISHI; Haruto SASAKI

    2006-01-01

    Dynamic changes of sucrose, fructose, glucose contents and differences in activities of sucrose synthase, vacuolar invertase, and cell wall bound invertase in rice grain after flowering stage were studied under natural and high temperatures by using two japonica rice varieties Koshihikari and Sasanishiki. In rice grains, the sucrose synthase activity was higher than that of invertase, which was significantly correlated with starch accumulation rate, indicating that the sucrose synthase played an important role in sucrose degradation and starch synthesis. Under high temperature, the significant increase in grain sucrose content without any increase in fructose and glucose contents, suggested that the high temperature treatment enhanced sucrose accumulation, while diminished sucrose degradation in rice grains. Compared with the control plants, the decrease in activities of sucrose synthase, vacuolar invertase, and cell wall bound invertase with high temperature treated plants indicated that the deceleration of sucrose-degradation was related to the decrease in activities of sucrose synthase and invertase.

  9. Precipitation of aluminum nitride in a high strength maraging steel with low nitrogen content

    Energy Technology Data Exchange (ETDEWEB)

    Jeanmaire, G., E-mail: guillaume.jeanmaire@univ-lorraine.fr [Institut Jean Lamour, UMR 7198 CNRS-Université de Lorraine, 54011 Nancy Cedex (France); Aubert and Duval, BP1, 63770 Les Ancizes (France); Dehmas, M.; Redjaïmia, A. [Institut Jean Lamour, UMR 7198 CNRS-Université de Lorraine, 54011 Nancy Cedex (France); Puech, S. [Aubert and Duval, BP1, 63770 Les Ancizes (France); Fribourg, G. [Snecma Gennevilliers, 171 Boulevard de Valmy-BP 31, 92702 Colombes (France)

    2014-12-15

    In the present work, aluminum nitride (AlN) precipitation was investigated in a X23NiCoCrMoAl13-6-3 maraging steel with low nitrogen content (wt.% N = 5.5 ppm). A reliable and robust automatic method by scanning electron microscopy observations coupled with energy dispersive X-ray spectroscopy was developed for the quantification of AlN precipitates. The first stage was to identify the solvus temperature and to develop a heat treatment able to dissolve the AlN precipitates. The experimental determination of equilibrium conditions and solvus temperature show good agreement with ThermoCalc® simulation. Then, from this AlN-free state, the cooling rate, isothermal holding time and temperature were the subject of an intensive investigation in the austenite region of this maraging steel. In spite of the high temperatures used during heat treatments, the growth kinetic of the largest AlN precipitates (> 1 μm) is slow. The cooling rate has a major effect on the size and the number density of AlN due to a higher driving force for nucleation at low temperatures. At last, quenching prior to isothermal annealing at high temperatures leads to fine and dense AlN precipitation, resulting from the martensite to austenite transformation. Experimental results will be discussed and compared with kinetic data obtained with the mobility database MobFe2 implemented in Dictra® software. - Highlights: • Slow dissolution kinetic of AlN precipitates due to both their large size and small chemical driving force • Significant effects of cooling rate prior isothermal heat treatment, holding time and temperature on AlN precipitation • Size of AlN precipitates can be reduced by quenching prior isothermal holding. • Fine precipitation of AlN related to the α → γ transformation.

  10. High boron content carboranyl-functionalized aryl ether derivatives displaying photoluminescent properties.

    Science.gov (United States)

    Lerouge, Frédéric; Viñas, Clara; Teixidor, Francesc; Núñez, Rosario; Abreu, Arturo; Xochitiotzi, Elba; Santillan, Rosa; Farfán, Norberto

    2007-05-21

    The reaction of alpha,alpha'-bis(3,5-bis(bromomethyl)phenoxy-p-xylene (3) with 4 equiv of the monolithium salt of 1-Ph-1,2-C2B10H11 or 1-Me-1,2-C2B10H11 gave the corresponding neutral carboranyl-functionalized aryl ether derivatives closo-4 and closo-5, respectively. These compounds contain four closo clusters that were degraded using basic conditions with KOH in EtOH, affording the corresponding nido-6 and nido-7 as potassium salts. Nido species were also isolated with tetramethylammonium as cation giving compounds nido-8 and nido-9 in good yield. The potassium salts showed good solubility in water and polar solvents. All these compounds were characterized by 1H, 11B and 13C NMR spectroscopy and UV-vis. The electronic data in different solvents indicated a solvatochromic shift for all compounds and a red shift of the absorption maxima for the nido species with respect to the closo derivatives. These neutral and anionic carboranyl-functionalized aryl ether derivatives represent a new family of high boron content luminescent compounds that show strong fluorescence emission in different solvents at room temperature. This phenomenon is very interesting considering the fact that none of the precursors have such a property. The fluorescence emission depends on the cluster substituent (Ph or Me) and the solvent polarity. Additionally, the fluorescence emission intensity was clearly dependent on the solvent polarity; the closo species showed strongest fluorescence intensities in the non-polar solvents, while anionic species were highly emissive in polar solvents.

  11. Label-free high-throughput detection and content sensing of individual droplets in microfluidic systems.

    Science.gov (United States)

    Yesiloz, Gurkan; Boybay, Muhammed Said; Ren, Carolyn L

    2015-10-21

    This study reports a microwave-microfluidics integrated approach capable of performing droplet detection at high-throughput as well as content sensing of individual droplets without chemical or physical intrusion. The sensing system consists of a custom microwave circuitry and a spiral-shaped microwave resonator that is integrated with microfluidic chips where droplets are generated. The microwave circuitry is very cost effective by using off-the-shelf components only. It eliminates the need for bulky benchtop equipment, and provides a compact, rapid and sensitive tool compatible for Lab-on-a-Chip (LOC) platforms. To evaluate the resonator's sensing capability, it was first applied to differentiate between single-phase fluids which are aqueous solutions with different concentrations of glucose and potassium chloride respectively by measuring its reflection coefficient as a function of frequency. The minimum concentration assessed was 0.001 g ml(-1) for potassium chloride and 0.01 g ml(-1) for glucose. In the droplet detection experiments, it is demonstrated that the microwave sensor is able to detect droplets generated at as high throughput as 3.33 kHz. Around two million droplets were counted over a period of ten minutes without any missing. For droplet sensing experiments, pairs of droplets that were encapsulated with biological materials were generated alternatively in a double T-junction configuration and clearly identified by the microwave sensor. The sensed biological materials include fetal bovine serum, penicillin antibiotic mixture, milk (2% mf) and d-(+)-glucose. This system has significant advantages over optical detection methods in terms of its cost, size and compatibility with LOC settings and also presents significant improvements over other electrical-based detection techniques in terms of its sensitivity and throughput.

  12. Continuous-flow sorting of microalgae cells based on lipid content by high frequency dielectrophoresis

    Directory of Open Access Journals (Sweden)

    Doug Redelman

    2016-08-01

    Full Text Available This paper presents a continuous-flow cell screening device to isolate and separate microalgae cells (Chlamydomonas reinhardtii based on lipid content using high frequency (50 MHz dielectrophoresis. This device enables screening of microalgae due to the balance between lateral DEP forces relative to hydrodynamic forces. Positive DEP force along with amplitude-modulated electric field exerted on the cells flowing over the planar interdigitated electrodes, manipulated low-lipid cell trajectories in a zigzag pattern. Theoretical modelling confirmed cell trajectories during sorting. Separation quantification and sensitivity analysis were conducted with time-course experiments and collected samples were analysed by flow cytometry. Experimental testing with nitrogen starveddw15-1 (high-lipid, HL and pgd1 mutant (low-lipid, LL strains were carried out at different time periods, and clear separation of the two populations was achieved. Experimental results demonstrated that three populations were produced during nitrogen starvation: HL, LL and low-chlorophyll (LC populations. Presence of the LC population can affect the binary separation performance. The continuous-flow micro-separator can separate 74% of the HL and 75% of the LL out of the starting sample using a 50 MHz, 30 voltages peak-to-peak AC electric field at Day 6 of the nitrogen starvation. The separation occurred between LL (low-lipid: 86.1% at Outlet # 1 and LC (88.8% at Outlet # 2 at Day 9 of the nitrogen starvation. This device can be used for onsite monitoring; therefore, it has the potential to reduce biofuel production costs

  13. Mapping whole-brain activity with cellular resolution by light-sheet microscopy and high-throughput image analysis (Conference Presentation)

    Science.gov (United States)

    Silvestri, Ludovico; Rudinskiy, Nikita; Paciscopi, Marco; Müllenbroich, Marie Caroline; Costantini, Irene; Sacconi, Leonardo; Frasconi, Paolo; Hyman, Bradley T.; Pavone, Francesco S.

    2016-03-01

    Mapping neuronal activity patterns across the whole brain with cellular resolution is a challenging task for state-of-the-art imaging methods. Indeed, despite a number of technological efforts, quantitative cellular-resolution activation maps of the whole brain have not yet been obtained. Many techniques are limited by coarse resolution or by a narrow field of view. High-throughput imaging methods, such as light sheet microscopy, can be used to image large specimens with high resolution and in reasonable times. However, the bottleneck is then moved from image acquisition to image analysis, since many TeraBytes of data have to be processed to extract meaningful information. Here, we present a full experimental pipeline to quantify neuronal activity in the entire mouse brain with cellular resolution, based on a combination of genetics, optics and computer science. We used a transgenic mouse strain (Arc-dVenus mouse) in which neurons which have been active in the last hours before brain fixation are fluorescently labelled. Samples were cleared with CLARITY and imaged with a custom-made confocal light sheet microscope. To perform an automatic localization of fluorescent cells on the large images produced, we used a novel computational approach called semantic deconvolution. The combined approach presented here allows quantifying the amount of Arc-expressing neurons throughout the whole mouse brain. When applied to cohorts of mice subject to different stimuli and/or environmental conditions, this method helps finding correlations in activity between different neuronal populations, opening the possibility to infer a sort of brain-wide 'functional connectivity' with cellular resolution.

  14. Sex Education in Connecticut High Schools: Teachers' Reports of Content and Importance Ratings According to the SIECUS Guidelines

    Science.gov (United States)

    Obloj, Wallace; Lynn, Donna

    2010-01-01

    The purpose of this study was to examine Connecticut teachers' reports of the sex education content taught to high school students as well as teachers' reports of the degree of importance for Connecticut high school students to understand according to the SIECUS Guidelines. The data revealed that participants (N=125) reported teaching 72% of the…

  15. New high boron content polyborane precursors to advanced ceramic materials: New syntheses, new applications

    Science.gov (United States)

    Guron, Marta

    There is a need for new synthetic routes to high boron content materials for applications as polymeric precursors to ceramics, as well as in neutron shielding and potential medical applications. To this end, new ruthenium-catalyzed olefin metathesis routes have been devised to form new complex polyboranes and polymeric species. Metathesis of di-alkenyl substituted o-carboranes allowed the synthesis of ring-closed products fused to the carborane cage, many of which are new compounds and one that offers a superior synthetic method to one previously published. Acyclic diene metathesis of di-alkenyl substituted m-carboranes resulted in the formation of new main-chain carborane-containing polymers of modest molecular weights. Due to their extremely low char yields, and in order to explore other metathesis routes, ring opening metathesis polymerization (ROMP) was used to generate the first examples of poly(norbornenyl- o-carboranes). Monomer synthesis was achieved via a two-step process, incorporating Ti-catalyzed hydroboration to make 6-(5-norbornenyl)-decaborane, followed by alkyne insertion in ionic liquid media to achieve 1,2-R2 -3-norbornenyl o-carborane species. The monomers were then polymerized using ROMP to afford several examples of poly(norbornenyl- o-carboranes) with relatively high molecular weights. One such polymer, [1-Ph, 3-(=CH2-C5H7-CH2=)-1,2-C 2B10H10]n, had a char yield very close to the theoretical char yield of 44%. Upon random copolymerization with poly(6-(5-norbornenyl) decaborane), char yields significantly increased to 80%, but this number was well above the theoretical value implicating the formation of a boron-carbide/carbon ceramic. Finally, applications of polyboranes were explored via polymer blends toward the synthesis of ceramic composites and the use of polymer precursors as reagents for potential ultra high temperature ceramic applications. Upon pyrolysis, polymer blends of poly(6-(5-norbornenyl)-decaborane) and poly

  16. Architected Cellular Materials

    Science.gov (United States)

    Schaedler, Tobias A.; Carter, William B.

    2016-07-01

    Additive manufacturing enables fabrication of materials with intricate cellular architecture, whereby progress in 3D printing techniques is increasing the possible configurations of voids and solids ad infinitum. Examples are microlattices with graded porosity and truss structures optimized for specific loading conditions. The cellular architecture determines the mechanical properties and density of these materials and can influence a wide range of other properties, e.g., acoustic, thermal, and biological properties. By combining optimized cellular architectures with high-performance metals and ceramics, several lightweight materials that exhibit strength and stiffness previously unachievable at low densities were recently demonstrated. This review introduces the field of architected materials; summarizes the most common fabrication methods, with an emphasis on additive manufacturing; and discusses recent progress in the development of architected materials. The review also discusses important applications, including lightweight structures, energy absorption, metamaterials, thermal management, and bioscaffolds.

  17. Development of CAPS markers to identify Indian tea (Camellia sinensis) clones with high catechin content.

    Science.gov (United States)

    Elangbam, M; Misra, A K

    2016-06-10

    Tea leaves are rich in plant secondary phenolics, especially flavonoids. Catechins are considered to be the most valuable flavonoids, and the catechin content in tea is an important trait for determining its quality. We have developed cleaved amplified polymorphic sequence (CAPS)-based markers for evaluating total catechin content that target two important secondary metabolite pathway genes, PAL (phenylalanine ammonia-lyase) and CHS (chalcone synthase). Catechin content levels in the tea samples tested ranged from 9 to 33 mg/mg. The CAPS technique identified clones with the homozygous profile PRc1, which has relatively lower catechin content than clones with the heterozygous profile PRc2. A significant difference (t = 16.85) in the level of catechin content was also detected between heterozygotes and homozygotes in the tea seed stock TS379. We found a polynomial relationship between the marker developed for CHS2 and catechin content in these tea samples with R2 = 0.9788. Moreover, PAL has less of a relationship with catechin content. Therefore, we recommend tea clones with heterozygous CAPS profiles for the gene CHS2+RcaI for the further improvement in these clones.

  18. A possible role for extra-cellular ATP in plant responses to high frequency, low amplitude electromagnetic field.

    Science.gov (United States)

    Roux, David; Faure, Catherine; Bonnet, Pierre; Girard, Sébastien; Ledoigt, Gérard; Davies, Eric; Gendraud, Michel; Paladian, Françoise; Vian, Alain

    2008-06-01

    In parallel to evoking the accumulation of stress-related transcripts, exposure to low level 900 MHz EMF affected the levels of ATP, the main energy molecule of the cell. Its concentration dropped rapidly (27% after 30 min) in response to EMF exposure, along with a 18% decrease in the adenylate energy charge (AEC), a good marker of cell energy status. One could interpret this decrease in ATP and AEC in a classical way, i.e., as the result of an increase in cellular energy usage, but recent work brings exciting new insights in pointing out a signalling function for ATP, especially in the stress physiology context where it could trigger both reactive oxygen species and calcium movement (this latter being involved in plant responses to EMF exposure). In this addendum, we discuss our results within this new perspective for ATP function.

  19. Highly differentiated cellular and circuit properties of infralimbic pyramidal neurons projecting to the periaqueductal gray and amygdala.

    Directory of Open Access Journals (Sweden)

    Ashley N Ferreira

    2015-04-01

    Full Text Available The infralimbic (IL cortex is a key node in an inter-connected network involved in fear and emotion processing. The cellular and circuit-level mechanisms whereby IL neurons receive, filter, and modulate incoming signals they project onward to diverse downstream nodes in this complex network remain poorly understood. Using the mouse as our model, we applied anatomical labeling strategies, brain slice electrophysiology, and focal activation of caged glutamate via laser scanning photostimulation (glu-LSPS for quantitative neurophysiological analysis of projectionally defined neurons in IL. Injection of retrograde tracers into the periaqueductal grey (PAG and basolateral amygdala (BLA was used to identify cortico-PAG (CP and cortico-BLA (CA neurons in IL. CP neurons were found exclusively in layer 5 of IL whereas CA neurons were detected throughout layer 2, 3 and 5 of IL. We also identified a small percentage of IL neurons that project to both the PAG and the BLA. We found that layer 5 CP neurons have a more extensive dendritic structure compared to layer 5 CA neurons. Neurophysiological recordings performed on retrogradely labeled neurons in acute brain slice showed that CP and CA neurons in IL could be broadly classified in two groups: neuronal resonators and non-resonators. Layer 2 CA neurons were the only class that was exclusively non-resonating. CP, CA, and CP/CA neurons in layers 3 and 5 of IL consisted of heterogeneous populations of resonators and non-resonators showing that projection target is not an exclusive predictor of intrinsic physiology. Circuit mapping using glu-LSPS revealed that the strength and organization of local excitatory and inhibitory inputs were stronger to CP compared to CA neurons in IL. Together, our results establish an organizational scheme linking cellular neurophysiology with microcircuit parameters of defined neuronal subclasses in IL that send descending commands to subcortical structures involved in fear

  20. Quantification of patient-derived 3D cancer spheroids in high-content screening images

    Science.gov (United States)

    Kang, Mi-Sun; Rhee, Seon-Min; Seo, Ji-Hyun; Kim, Myoung-Hee

    2017-02-01

    We present a cell image quantification method for image-based drug response prediction from patient-derived glioblastoma cells. Drug response of each person differs at the cellular level. Therefore, quantification of a patient-derived cell phenotype is important in drug response prediction. We performed fluorescence microscopy to understand the features of patient-derived 3D cancer spheroids. A 3D cell culture simulates the in-vivo environment more closely than 2D adherence culture, and thus, allows more accurate cell analysis. Furthermore, it allows assessment of cellular aggregates. Cohesion is an important feature of cancer cells. In this paper, we demonstrate image-based quantification of cellular area, fluorescence intensity, and cohesion. To this end, we first performed image stitching to create an image of each well of the plate with the same environment. This image shows colonies of various sizes and shapes. To automatically detect the colonies, we used an intensity based classification algorithm. The morphological features of each cancer cell colony were measured. Next, we calculated the location correlation of each colony that is appeal of the cell density in the same well environment. Finally, we compared the features for drug-treated and untreated cells. This technique could potentially be applied for drug screening and quantification of the effects of the drugs.

  1. Optimization of conditions to achieve high content of gamma amino butyric acid in germinated black rice, and changes in bioactivities

    Directory of Open Access Journals (Sweden)

    Chaiyavat CHAIYASUT

    Full Text Available Abstract The present study estimated the optimum germination conditions to achieve high content of Gamma-amino butyric acid (GABA and other phytochemicals in Thai black rice cultivar Kum Payao (BR. The Box–Behnken design of response surface methodology was employed to optimize the germination conditions. The changes in the GABA, phytochemical content, impact of salt, and temperature stress variation on phytochemical content, and stability of GABA were studied. The results showed that 12 h of soaking at pH 7, followed by 36 h of germination was the optimum condition to achieve maximum GABA content (0.2029 mg/g of germinated BR (GBR. The temperature (8 and 30 °C, and salt (50-200 mM NaCl content affected the phytochemicals of GBR, especially GABA, and anthocyanins. Obviously, the antioxidant capability, and enzyme (α-amylase and α-glucosidase inhibiting nature of BR was significantly (P < 0.001 increased after germination. The storage of GBR at 4 °C significantly, preserved the GABA content (∼80% for 45 days. Primarily, the current study revealed the changes in phytochemical content, and bioactivity of Thai black rice cr. Kum Payao during germination. More studies should be carried out on pharmacological benefits of GABA-rich GBR.

  2. Effects of Nitrogen Content on the HAZ Softening of Ti-Containing High Strength Steels Manufactured by Accelerated Cooling

    Energy Technology Data Exchange (ETDEWEB)

    Bang, Kook-soo; Jung, Ho-shin; Park, Chan [Pukyong National University, Busan (Korea, Republic of)

    2017-03-15

    The effects of nitrogen content on the HAZ softening of Ti-containing high strength steels manufactured by accelerating cooling were investigated and interpreted in terms of the microstructures in the softening zone. Regardless of their content, all of the steels investigated showed a softened zone 9-10 mm wide. The minimum hardness in the zone, however, was different, with lower hardness in the higher nitrogen content steel. Microstructural observations of the steel showed that the amount of soft ferrite was increased in the zone with an increase of nitrogen content of the steel, suggesting that microstructural evolution in the HAZ is influenced by the nitrogen content. Measurements of TiN particles showed that the degree of particles coarsening in the HAZ was lower in the higher nitrogen content steel. Therefore, it is believed that finer TiN particles in the HAZ inhibit austenite grain growth more effectively, and lead to an accelerated ferrite transformation in higher nitrogen content steel, resulting in a higher amount of soft ferrite microstructure in the softened zone.

  3. Identification of differentially expressed genes in longissimus muscle of pigs with high and low intramuscular fat content using RNA sequencing.

    Science.gov (United States)

    Lim, K S; Lee, K T; Park, J E; Chung, W H; Jang, G W; Choi, B H; Hong, K C; Kim, T H

    2017-04-01

    Intramuscular fat (IMF) content in pork is an important element of consumer preference and is positively correlated with meat quality, including tenderness and juiciness. With advances in RNA sequencing technologies, transcriptome-related differences can be associated with specific traits in animals. The objective of this study was to investigate differentially expressed genes (DEGs) closely related to IMF content in porcine longissimus muscle using RNA sequencing. A total of 107 Berkshire pigs were used for IMF content measurements, and significant differences between extremely high (H, n = 3) and low (L, n = 3) IMF content groups were found (P change ≥2). Functional analyses with DEGs revealed that lipid metabolism (SCD and FASN) was one of the significant biological processes related to IMF content determination. In addition, we found that DEGs related to muscle regeneration (MYOG and VEGFA) and extracellular matrix (COL1A1, COL1A2, COL5A1, COL14A1 and COL15A1) were changed among individuals with extreme IMF contents. These results will aid in understanding the regulation of IMF content in pigs.

  4. Rapid solid-phase microwave synthesis of highly photoluminescent nitrogen-doped carbon dots for Fe3+ detection and cellular bioimaging

    Science.gov (United States)

    He, Guili; Xu, Minghan; Shu, Mengjun; Li, Xiaolin; Yang, Zhi; Zhang, Liling; Su, Yanjie; Hu, Nantao; Zhang, Yafei

    2016-09-01

    Recently, carbon dots (CDs) have been playing an increasingly important role in industrial production and biomedical field because of their excellent properties. As such, finding an efficient method to quickly synthesize a large scale of relatively high purity CDs is of great interest. Herein, a facile and novel microwave method has been applied to prepare nitrogen doped CDs (N-doped CDs) within 8 min using L-glutamic acid as the sole reaction precursor in the solid phase condition. The as-prepared N-doped CDs with an average size of 1.64 nm are well dispersed in aqueous solution. The photoluminescence of N-doped CDs is pH-sensitive and excitation-dependent. The N-doped CDs show a strong blue fluorescence with relatively high fluorescent quantum yield of 41.2%, which remains stable even under high ionic strength. Since the surface is rich in oxygen-containing functional groups, N-doped CDs can be applied to selectively detect Fe3+ with the limit of detection of 10-5 M. In addition, they are also used for cellular bioimaging because of their high fluorescent intensity and nearly zero cytotoxicity. The solid-phase microwave method seems to be an effective strategy to rapidly obtain high quality N-doped CDs and expands their applications in ion detection and cellular bioimaging.

  5. The application of content based teaching method in English writing in Senior High School

    Institute of Scientific and Technical Information of China (English)

    李素霞

    2016-01-01

    Content-based instruction (CBI) refers to an approach of subject integration to language teaching in which the teaching is organized around the subject or information that students will acquire. Based on the deficiency of the previous studies, the thesis proposes a Content-Based Instruction, in order to improve the quality of English Writing Teaching in senior school and students' actual level of writing.

  6. Effect of Nitrogen Content and Cooling Rate on Transformation Characteristics and Mechanical Properties for 600 MPa High Strength Rebar

    Science.gov (United States)

    Zhang, Jing; Wang, Fu-ming; Li, Chang-rong; Yang, Zhan-bing

    2016-10-01

    To obtain appropriate chemical composition and thermo-mechanical schedules for processing the V-N microalloyed 600 MPa grade high strength rebar, the microstructure analysis during dynamic continuous cooling and tensile tests of three experimental steels with different nitrogen contents were conducted. The results show that increasing nitrogen content promotes ferrite transformation and broadens the bainite transformation interval, when the nitrogen content is in the range of 0.019-0.034 mass%. Meanwhile, the martensite start temperatures decrease and the minimal cooling rate to form martensite increases. To achieve a good combination of strength and ductility, the cooling rates should be controlled in the range of 0.5-3°C/s, leading to the microstructure of ferrite, pearlite and less than 10% bainite (volume fraction). Furthermore, all the experimental steels satisfy the performance requirement of 600 MPa grade rebar and the rebar with nitrogen content of 0.034 mass% shows the highest strength through systematically comparative investigation.

  7. PHASE SEPARATION IN BIMODAL MOLECULAR WEIGHT HIGH DENSITY POLYETHYLENE WITH DIFFERING BRANCH CONTENTS BY MOLECULAR DYNAMICS AND MESODYN SIMULATION

    Institute of Scientific and Technical Information of China (English)

    Zhi-jie Zhang; Zhong-yuan Lu; Ze-sheng Li

    2009-01-01

    The phase behavior of bimodal molecular weight high density polyethylene (BHDPE) in solid state was investigated. Hildebrand solubility parameters (δ) were calculated for the models of blends of higher molecular weight branch polyethylene (HBPE) with different branch contents and lower molecular weight linear polyethylene (LLPE), by using molecular dynamics (MD) simulations. These δ values were then used to calculate the corresponding Flory-Huggins interaction parameter (χ) between HBPE and LLPE models. In order to better understand the compatibility between LLPE and various HBPE, Mesodyn simulations were used to show the density profiles of the blends of LLPE with various HBPE at different compositions. The results indicated that the phase behavior of BHDPE was influenced by both the global branch content of the system and the local branch content, I.e., the branch content of HBPE.

  8. Validation of a high-content screening assay using whole-well imaging of transformed phenotypes.

    Science.gov (United States)

    Ramirez, Christina N; Ozawa, Tatsuya; Takagi, Toshimitsu; Antczak, Christophe; Shum, David; Graves, Robert; Holland, Eric C; Djaballah, Hakim

    2011-06-01

    Automated microscopy was introduced two decades ago and has become an integral part of the discovery process as a high-content screening platform with noticeable challenges in executing cell-based assays. It would be of interest to use it to screen for reversers of a transformed cell phenotype. In this report, we present data obtained from an optimized assay that identifies compounds that reverse a transformed phenotype induced in NIH-3T3 cells by expressing a novel oncogene, KP, resulting from fusion between platelet derived growth factor receptor alpha (PDGFRα) and kinase insert domain receptor (KDR), that was identified in human glioblastoma. Initial image acquisitions using multiple tiles per well were found to be insufficient as to accurately image and quantify the clusters; whole-well imaging, performed on the IN Cell Analyzer 2000, while still two-dimensional imaging, was found to accurately image and quantify clusters, due largely to the inherent variability of their size and well location. The resulting assay exhibited a Z' value of 0.79 and a signal-to-noise ratio of 15, and it was validated against known effectors and shown to identify only PDGFRα inhibitors, and then tested in a pilot screen against a library of 58 known inhibitors identifying mostly PDGFRα inhibitors as reversers of the KP induced transformed phenotype. In conclusion, our optimized and validated assay using whole-well imaging is robust and sensitive in identifying compounds that reverse the transformed phenotype induced by KP with a broader applicability to other cell-based assays that are challenging in HTS against chemical and RNAi libraries.

  9. Reactivity and burnability of cement raw materials witt high manganese content

    Directory of Open Access Journals (Sweden)

    Puertas, F.

    1988-06-01

    Full Text Available It has been verified that high manganese content in raw mixes causes alters the mineralogical composition of clinkers. New phases like 2 CaO.Mn02 and Ca2AIMnO5 are developed and C3A formation can be inhibited. Manganese is a flux similar to iron in the traditional raw mixes. The presence of manganese will modify the expressions of the lime saturation factor (LSF, silica modulus (Ms and aluminum modulus (Mf. This has as consequence an increase of the proportion of interstitial phase in the obtained clinkers.

    Se ha comprobado que contenidos elevados de manganeso en el crudo provoca una modificación significativa en la composición mineralógica del clinker. Se desarrollan las fases no tradicionales 2 CaO.Mn02 y Ca2AIMnO5 , y se puede llegar, incluso, a inhibir la formación de alumínate tricálcico. El manganeso tiene un comportamiento fundente muy similar al jugado por el hierro en los crudos tradicionales, por lo que la incorporación de óxidos de manganeso al crudo produce una alteración significativa en los módulos tradicionales (LSF, Ms y Mf. Esto tiene como consecuencia inmediata un gran incremento en la proporción de fase intersticial en los clínkeres producidos.

  10. Investigation of possibilities for high heavy metal content sludges utilization by incorporating them in concrete products

    Directory of Open Access Journals (Sweden)

    Simeonova A.

    2006-01-01

    Full Text Available The safe removal of sludge, obtained during the surface treatment of different metal products, is a serious environmental problem. These sludges are usually characterized by a high content of heavy metals (Pb, Cu, Ni, Zn, Cr, Cd, Mn, low quality and are obtained in many small industrial units in the whole country, which makes their centralized treatment difficult. In world practice, different methods are used for component fixation of such sludge, in the aim to prevent leaching of the metals causing pollution of the soil and underground water. The aim of the recent work is to prepare the sludge in a form of light (keramzit fillers by preliminary treatment with binding substances and to introduce them in non supporting concrete products -