Sample records for total cell quantities

  1. Proteomic analysis of cellular soluble proteins from human bronchial smooth muscle cells by combining nondenaturing micro 2DE and quantitative LC-MS/MS. 1. Preparation of more than 4000 native protein maps.

    PubMed

    Jin, Ya; Zhang, Jun; Yuan, Qi; Manabe, Takashi; Tan, Wen

    2015-08-01

    Soluble proteins of human bronchial smooth muscle cells (HBSMC) were separated by nondenaturing micro 2DE and a 30 mm × 40 mm area of the CBB-stained slab gel (1.0 mm thick) was cut into 1.1 mm × 1.1 mm squares, then the proteins in the 972 gel pieces (squares) were applied to quantitative LC-MS/MS. Grid-cutting of the gel was employed to; (i) ensure the total analysis of the proteins in the area, (ii) standardize the conditions of analysis by LC-MS/MS, (iii) reconstruct the protein distribution patterns from the quantity data [1]. Totally 4323 proteins were identified in successfully analyzed 967 squares and the quantity distribution of each was reconstructed as a color density pattern (a native protein map). The quantity of the proteins distributed from 3.6% to 1 × 10(-5) % of the total protein quantity in the grid area. Each protein map was characterized by several features, including the position of quantity peak square, number of detected squares, and degree of concentration (focused or dispersed). About 4% of the proteins were detected in 100 or more squares, suggesting that they might be ubiquitous and interacting with other proteins. In contrast, many proteins showed more concentrated quantity distribution and the quantity peak positions of 565 proteins with a defined degree of concentration were summarized into a quantity peak map. These results for the first time visualized the distribution patterns of cellular proteins on a nondenaturing 2D gel. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Effect of Increased Cyclic AMP Concentration on Muscle Protein Synthesis and Beta-Adrenergic Receptor Expression in Chicken Skeletal Muscle Cells in Culture

    NASA Technical Reports Server (NTRS)

    Young, R. B.; Vaughn, J. R.; Bridge, K. Y.; Smith, C. K.

    1998-01-01

    Analogies of epinephrine are known to cause hypertrophy of skeletal muscle when fed to animals. These compounds presumably exert their physiological action through interaction with the P-adrenergic receptor. Since the intracellular signal generated by the Beta-adrenergic receptor is cyclic AMP (cAMP), experiments were initiated in cell culture to determine if artificial elevation of cAMP by treatment with forskolin would alter muscle protein metabolism and P-adrenergic receptor expression. Chicken skeletal muscle cells after 7 days in culture were treated with 0.2-30 micrometers forskolin for a total of three days. At the end of the treatment period, both the concentration of cAMP and the quantity of myosin heavy chain (MHC) were measured. Concentration of cAMP in forskolin-treated cells increased up to 10-fold in a dose dependent manner. In contrast, the quantity of MHC was increased approximately 50% above control cells at 0.2 micrometers forskolin, but exhibited a gradual decline at higher levels of forskolin so that the quantity of MHC in cells treated with 30 micrometers forskolin was not significantly different from controls. Curiously, the intracellular concentration of cAMP which elicited the maximum increase in the quantity of MHC was only 40% higher than cAMP concentration in control cells.

  3. The kinetics of translocation and cellular quantity of protein kinase C in human leukocytes are modified during spaceflight

    NASA Technical Reports Server (NTRS)

    Hatton, J. P.; Gaubert, F.; Lewis, M. L.; Darsel, Y.; Ohlmann, P.; Cazenave, J. P.; Schmitt, D.

    1999-01-01

    Protein kinase C (PKC) is a family of serine/threonine kinases that play an important role in mediating intracellular signal transduction in eukaryotes. U937 cells were exposed to microgravity during a space shuttle flight and stimulated with a radiolabeled phorbol ester ([3H]PDBu) to both specifically label and activate translocation of PKC from the cytosol to the particulate fraction of the cell. Although significant translocation of PKC occurred at all g levels, the kinetics of translocation in flight were significantly different from those on the ground. In addition, the total quantity of [3H]PDBu binding PKC was increased in flight compared to cells at 1 g on the ground, whereas the quantity in hypergravity (1.4 g) was decreased with respect to 1 g. Similarly, in purified human peripheral blood T cells the quantity of PKCdelta varied in inverse proportion to the g level for some experimental treatments. In addition to these novel findings, the results confirm earlier studies which showed that PKC is sensitive to changes in gravitational acceleration. The mechanisms of cellular gravisensitivity are poorly understood but the demonstrated sensitivity of PKC to this stimulus provides us with a useful means of measuring the effect of altered gravity levels on early cell activation events.

  4. Experimental Techniques for Thermodynamic Measurements of Ceramics

    NASA Technical Reports Server (NTRS)

    Jacobson, Nathan S.; Putnam, Robert L.; Navrotsky, Alexandra

    1999-01-01

    Experimental techniques for thermodynamic measurements on ceramic materials are reviewed. For total molar quantities, calorimetry is used. Total enthalpies are determined with combustion calorimetry or solution calorimetry. Heat capacities and entropies are determined with drop calorimetry, differential thermal methods, and adiabatic calorimetry . Three major techniques for determining partial molar quantities are discussed. These are gas equilibration techniques, Knudsen cell methods, and electrochemical techniques. Throughout this report, issues unique to ceramics are emphasized. Ceramic materials encompass a wide range of stabilities and this must be considered. In general data at high temperatures is required and the need for inert container materials presents a particular challenge.

  5. Measurement of Total Site Mercury Emissions from a Chlor-Alkali Plant Using Ultraviolet Differential Optical Absorption Spectroscopy and Cell Room Roof-Vent Monitoring

    EPA Science Inventory

    Mercury-cell chlor-alkali plants can emit significant quantities of fugitive elemental mercury vapor to the air as part of production operations and maintenance activities. In the fall of 2006, the U.S. Environmental Protection Agency (EPA) conducted a measurement project at a ch...

  6. Stilbene production in cell cultures of Vitis vinifera L. cvs Red Globe and Michele Palieri elicited by methyl jasmonate.

    PubMed

    Santamaria, A R; Antonacci, D; Caruso, G; Cavaliere, C; Gubbiotti, R; Lagana, A; Valletta, A; Pasqua, G

    2010-09-01

    Cell cultures obtained from Vitis vinifera cvs Michele Palieri and Red Globe were cultured in order to stimulate stilbene production. In the calli, stilbene production peaked at day 22 of culture for both cultivars; the main compound was trans-piceid, followed by cis-piceid. Methyl jasmonate, which was added to cell suspensions in the first half of the exponential growth phase, enhanced stilbene accumulation, producing mainly trans-piceid and epsilon-viniferin. Other stilbenoids, though in lower quantities, were identified by liquid chromatography/positive electrospray mass spectrometry. epsilon-Viniferin and trans-resveratrol were the main compounds released into the culture medium. The total quantity of stilbenes was genotype dependent, with a better response found for the cv Red Globe.

  7. Beta-Adrenergic Receptor Expression in Muscle Cells

    NASA Technical Reports Server (NTRS)

    Young, Ronald B.; Bridge, K.; Vaughn, J. R.

    1999-01-01

    beta-adrenergic receptor (bAR) agonists presumably exert their physiological action on skeletal muscle cells through the bAR. Since the signal generated by the bAR is cyclic AMP (cAMP), experiments were initiated in primary chicken muscle cell cultures to determine if artificial elevation of intracellular cAMP by treatment with forskolin would alter the population of bAR expressed on the surface of muscle cells. Chicken skeletal muscle cells after 7 days in culture were employed for the experiments because muscle cells have attained a steady state with respect to muscle protein metabolism at this stage. Cells were treated with 0-10 uM forskolin for a total of three days. At the end of the 1, 2, and 3 day treatment intervals, the concentration of cAMP and the bAR population were measured. Receptor population was measured in intact muscle cell cultures as the difference between total binding of [H-3]CGP-12177 and non-specific binding of [H-3]CGP-12177 in the presence of 1 uM propranolol. Intracellular cAMP concentration was measured by radioimmunoassay. The concentration of cAMP in forskolin-treated cells increased up to 10-fold in a dose dependent manner. Increasing concentrations of forskolin also led to an increase in (beta)AR population, with a maximum increase of approximately 50% at 10 uM. This increase in (beta)AR population was apparent after only 1 day of treatment, and the pattern of increase was maintained for all 3 days of the treatment period. Thus, increasing the intracellular concentration of cAMP leads to up-regulation of (beta)AR population. Clenbuterol and isoproterenol gave similar effects on bAR population. The effect of forskolin on the quantity and apparent synthesis rate of the heavy chain of myosin (mhc) were also investigated. A maximum increase of 50% in the quantity of mhc was observed at 0.2 UM forskolin, but higher concentrations of forskolin reduced the quantity of mhc back to control levels.

  8. Effect of a syringe aspiration technique versus a mechanical suction technique and use of N-butylscopolammonium bromide on the quantity and quality of bronchoalveolar lavage fluid samples obtained from horses with the summer pasture endophenotype of equine asthma.

    PubMed

    Bowser, Jacquelyn E; Costa, Lais R R; Rodil, Alba U; Lopp, Christine T; Johnson, Melanie E; Wills, Robert W; Swiderski, Cyprianna E

    2018-03-01

    OBJECTIVE To evaluate the effect of 2 bronchoalveolar lavage (BAL) sampling techniques and the use of N-butylscopolammonium bromide (NBB) on the quantity and quality of BAL fluid (BALF) samples obtained from horses with the summer pasture endophenotype of equine asthma. ANIMALS 8 horses with the summer pasture endophenotype of equine asthma. PROCEDURES BAL was performed bilaterally (right and left lung sites) with a flexible videoendoscope passed through the left or right nasal passage. During lavage of the first lung site, a BALF sample was collected by means of either gentle syringe aspiration or mechanical suction with a pressure-regulated wall-mounted suction pump. The endoscope was then maneuvered into the contralateral lung site, and lavage was performed with the alternate fluid retrieval technique. For each horse, BAL was performed bilaterally once with and once without premedication with NBB (21-day interval). The BALF samples retrieved were evaluated for volume, total cell count, differential cell count, RBC count, and total protein concentration. RESULTS Use of syringe aspiration significantly increased total BALF volume (mean volume increase, 40 mL [approx 7.5% yield]) and decreased total RBC count (mean decrease, 142 cells/μL), compared with use of mechanical suction. The BALF nucleated cell count and differential cell count did not differ between BAL procedures. Use of NBB had no effect on BALF retrieval. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that retrieval of BALF by syringe aspiration may increase yield and reduce barotrauma in horses at increased risk of bronchoconstriction and bronchiolar collapse. Further studies to determine the usefulness of NBB and other bronchodilators during BAL procedures in horses are warranted.

  9. Beta-Adrenergic Receptor Population is Up-Regulated in Chicken Skeletal Muscle Cells Treated with Forskolin

    NASA Technical Reports Server (NTRS)

    Bridge, K. Y.; Young, R. B.; Vaughn, J. R.

    1998-01-01

    Skeletal muscle hypertrophy is promoted by in vivo administration of beta-adrenergic receptor (betaAR) agonists. These compounds presumably exert their physiological action through the betaAR, and alterations in the population of betaAR could potentially change the ability of the cell to respond to the betaAR agonists. Since the intracellular chemical signal generated by the betaAR is cyclic AMP (cAMP), experiments were initiated in primary chicken muscle cell cultures to determine if artificial elevation of intracellular cAMP by treatment with forskolin would alter the population of functional betaAR expressed on the surface of muscle cells. Chicken skeletal muscle cells after 7 days in culture were employed for the experiments because muscle cells have attained a steady state with respect to muscle protein metabolism at this stage. Cells were treated with 0-10 microM forskolin for a total of three days. At the end of the 1, 2, and 3 day treatment intervals, the concentration of cAMP and the betaAR population were measured. Receptor population was measured in intact muscle cell cultures as the difference between total binding of [H-3]CGP-12177 and non-specific binding of [H-3]CGP-12177 in the presence of 1 microM propranolol. Intracellular cAMP concentration was measured by radioimmunoassay. The concentration of cAMP in forskolin-treated cells increased up to 10-fold in a dose dependent manner. Increasing concentrations of forskolin also led to an increase in betaAR population, with a maximum increase of approximately 50% at 10 microM. This increase in PAR population was apparent after only 1 day of treatment, and the pattern of increase was maintained for all 3 days of the treatment period. Thus, increasing the intracellular concentration of cAMP leads to up-regulation of betaAR population. The effect of forskolin on the quantity and apparent synthesis rate of the heavy chain of myosin (mhc) were also investigated. A maximum increase of 50% in the quantity of mhc was observed at 0.2 microM forskolin, but higher concentrations of forskolin reduced the quantity of mhc back to control levels.

  10. The generation of pollution-free electrical power from solar energy.

    NASA Technical Reports Server (NTRS)

    Cherry, W. R.

    1971-01-01

    Projections of the U.S. electrical power demands over the next 30 years indicate that the U.S. could be in grave danger from power shortages, undesirable effluence, and thermal pollution. An appraisal of nonconventional methods of producing electrical power is conducted, giving particular attention to the conversion of solar energy into commercial quantities of electrical power by solar cells. It is found that 1% of the land area of the 48 states could provide the total electrical power requirements of the U.S. in the year 1990. The ultimate method of generating vast quantities of electrical power would be from a series of synchronous satellites which beam microwave power back to earth to be used wherever needed. Present high manufacturing costs of solar cells could be substantially reduced by using massive automated techniques employing abundant low cost materials.

  11. Functional Characteristics of Tumor Associated Protein Spot14 and Interacting Proteins in Mouse Mammary Epithelial and Breast Cancer Cell Lines

    DTIC Science & Technology

    2012-03-01

    enhanced accumulation of total lipids evaluated by Bodipy staining and NMR analysis. A major finding in this report is that glycolytic and lipogenic enzyme...total lipid component using NMR Metabolomics showed significant increases in the quantity of intracellular (CH2)n and (CH3) acyl chains (i.e. fatty...Mass Spectrometry (GC-MS) methods were developed. GC-MS differs from NMR analysis of lipid fractions in that GC-MS distinguishes between fatty acids

  12. Variability in Beta-Adrenergic Receptor Population in Cultured Chicken Muscle Cells

    NASA Technical Reports Server (NTRS)

    Young, Ronald B; Bridge, Kristin Y.; Vaughn, Jeffrey R.

    1998-01-01

    Investigations into expression of the beta-adrenergic receptor (bAR) in chicken skeletal muscle cells in culture were initiated because several beta-adrenergic receptor agonists are known to increase skeletal muscle protein deposition in avian and mammalian species. During initial attempts to study the bAR population on the surface of chicken skeletal muscle cells, we observed a high degree of variability that was later found to be the result of using different batches of horse serum in the cell culture media. The separation between total binding and nonspecific binding in cells grown in two serum samples was approximately two-fold The number of nuclei within multinucleated myotubes was not significantly different in cells grown in the two serum samples. To investigate whether these two sera had an effect on coupling efficiency between bAR population and cAMP production, the ability of these cells to synthesize cAMP was also assessed. Despite the two-fold difference in receptor population, the ability of these cells to synthesize cAMP was not significantly different. Because of the possible link between bAR population and muscle protein, we also determined if the quantity of the major skeletal muscle protein, myosin, was affected by conditions that so drastically affected the bAR population. The quantity of myosin heavy chain was not significantly different.

  13. Controlled-rate freezer cryopreservation of highly concentrated peripheral blood mononuclear cells results in higher cell yields and superior autologous T-cell stimulation for dendritic cell-based immunotherapy.

    PubMed

    Buhl, Timo; Legler, Tobias J; Rosenberger, Albert; Schardt, Anke; Schön, Michael P; Haenssle, Holger A

    2012-11-01

    Availability of large quantities of functionally effective dendritic cells (DC) represents one of the major challenges for immunotherapeutic trials against infectious or malignant diseases. Low numbers or insufficient T-cell activation of DC may result in premature termination of treatment and unsatisfying immune responses in clinical trials. Based on the notion that cryopreservation of monocytes is superior to cryopreservation of immature or mature DC in terms of resulting DC quantity and immuno-stimulatory capacity, we aimed to establish an optimized protocol for the cryopreservation of highly concentrated peripheral blood mononuclear cells (PBMC) for DC-based immunotherapy. Cryopreserved cell preparations were analyzed regarding quantitative recovery, viability, phenotype, and functional properties. In contrast to standard isopropyl alcohol (IPA) freezing, PBMC cryopreservation in an automated controlled-rate freezer (CRF) with subsequent thawing and differentiation resulted in significantly higher cell yields of immature and mature DC. Immature DC yields and total protein content after using CRF were comparable with results obtained with freshly prepared PBMC and exceeded results of standard IPA freezing by approximately 50 %. While differentiation markers, allogeneic T-cell stimulation, viability, and cytokine profiles were similar to DC from standard freezing procedures, DC generated from CRF-cryopreserved PBMC induced a significantly higher antigen-specific IFN-γ release from autologous effector T cells. In summary, automated controlled-rate freezing of highly concentrated PBMC represents an improved method for increasing DC yields and autologous T-cell stimulation.

  14. [A comparative study on inducing non-homologous mesenchymal stem cells to differentiate into neural stem cells using non-homologous cerebrospinal fluid].

    PubMed

    Ren, Chao; Liu, Xiaoyun; Wan, Meirong; Geng, Deqin; Ge, Wei; Li, Jinmei; Zhang, Weiwei

    2013-12-01

    In order to set up a base for stem cells to be widely used in clinical medicine, we tried to optimize, in this study, the technique that induces human mesenchymal stem cells (hMSCs) to differentiate into neural stem cells by using cerebrospinal fluid (CSF) from the different groups. After the induction, presence of neural stem cells was confirmed with microscope observation, flow cytometry analysis, immunohistochemistry and fluorescent immunohistochemistry. At the same time, we also compared and analysed the data of the number of stem cells when it totally met the requirements for clinical treatment and the days required. At last, we confirmed that hMSCs could be induced to differentiate into neural stem cells, and that the number of cells totally met the requirements for clinical treatment. But there were some differences both in the number of cells and the days required. Among the groups, the group that marrow mesenchymal stem cells from patients own induced by CSF from healthy volunteers used the shortest time and the quantity of the cells was significantly higher than those of the others.

  15. Relationship between interphasic nucleolar organizer regions and growth rate in two neuroblastoma cell lines.

    PubMed Central

    Derenzini, M.; Pession, A.; Farabegoli, F.; Trerè, D.; Badiali, M.; Dehan, P.

    1989-01-01

    The relationship between the quantity of silver-stained interphasic nucleolar organizer regions (NORs) and nuclear synthetic activity, caryotype, and growth rate was studied in two established neuroblastoma cell lines (CHP 212 and HTB 10). Statistical analysis of silver-stained NORs revealed four times as many in CHP 212 cells compared with HTB 10 cells. No difference was observed in the ribosomal RNA synthesis between the two cell lines. The caryotype index was 1.2 for CHP 212 and 1.0 for HTB 10 cells. The number of chromosomes carrying NORs and the quantity of ribosomal genes was found to be the same for the two cell lines. Doubling time of CHP 212 cells was 20 hours compared with 54 hours for HTB 10 cells. In CHP 212 cells bindering of cell duplication by serum deprivation induced a progressive lowering (calculated at 48, 72, and 96 hours) of the quantity of silver-stained interphasic NORs. Recovery of duplication by new serum addition induced, after 24 hours, an increase of the quantity of silver-stained interphasic NORs up to control levels. In the light of available data, these results indicate that the quantity of interphasic NORs is strictly correlated only to the growth rate of the cell. Images Figure 2 Figure 3 Figure 4 PMID:2705511

  16. Microfluidic devices, systems, and methods for quantifying particles using centrifugal force

    DOEpatents

    Schaff, Ulrich Y.; Sommer, Gregory J.; Singh, Anup K.

    2015-11-17

    Embodiments of the present invention are directed toward microfluidic systems, apparatus, and methods for measuring a quantity of cells in a fluid. Examples include a differential white blood cell measurement using a centrifugal microfluidic system. A method may include introducing a fluid sample containing a quantity of cells into a microfluidic channel defined in part by a substrate. The quantity of cells may be transported toward a detection region defined in part by the substrate, wherein the detection region contains a density media, and wherein the density media has a density lower than a density of the cells and higher than a density of the fluid sample. The substrate may be spun such that at least a portion of the quantity of cells are transported through the density media. Signals may be detected from label moieties affixed to the cells.

  17. Anti-Inflammatory Effects of a Mixture of Lactic Acid Bacteria and Sodium Butyrate in Atopic Dermatitis Murine Model.

    PubMed

    Kim, Jeong A; Kim, Sung-Hak; Kim, In Sung; Yu, Da Yoon; Kim, Sung Chan; Lee, Seung Ho; Lee, Sang Suk; Yun, Cheol-Heui; Choi, In Soon; Cho, Kwang Keun

    2018-03-20

    Atopic dermatitis is a chronic and recurrent inflammatory skin disease. Recently, probiotics have been shown to suppress allergic symptoms through immunomodulatory responses. In the present study, combinatorial effects on allergic symptoms were identified in BALB/c mice fed with a mixture of four species of probiotics, Bifidobacterium lactis, Lactobacillus casei, Lactobacillus rhamnosus, and Lactobacillus plantarum, and sodium butyrate. Following sensitization with whey protein, the mice were challenged and divided into two groups: (1) mice administered with phosphate-buffered saline as a control and (2) mice administered with the probiotic mixture and sodium butyrate. Allergic symptoms were assessed by measuring ear thicknesses, serum histamine and IL-10 concentrations, and the quantities of leaked Evans blue. T cell differentiation was determined by analyzing the T cells groups in the mesenteric lymph nodes (MLNs) and spleen. To examine changes in the total gut microbiota, total fecal microflora was isolated, species identification was performed by DNA sequencing using Illumina MiSeq, and changes in intestinal beneficial bacteria were analyzed using quantitative polymerase chain reaction. Treatment with the probiotic mixture and sodium butyrate reduced ear thicknesses, the quantity of leaked Evans blue, and serum histamine values, while increasing serum IL-10 values. In the mouse model, the probiotic mixture and sodium butyrate increased Th1 and Treg cell differentiation in MLN and spleen tissues; the ratio of Firmicutes/Bacteroidetes, which is associated with reduction in allergic reactions; and microorganisms that lead to cell differentiation into Treg. These results suggest that the probiotic mixture and sodium butyrate can prevent and alleviate allergic symptoms.

  18. Liquid biopsy - Performance of the PAXgene® Blood ccfDNA Tubes for the isolation and characterization of cell-free plasma DNA from tumor patients.

    PubMed

    Schmidt, Bernd; Reinicke, Dana; Reindl, Iris; Bork, Ines; Wollschläger, Bettina; Lambrecht, Nina; Fleischhacker, Michael

    2017-06-01

    In most research laboratories the use of EDTA tubes for the isolation of plasma DNA from tumor patients is standard. Unfortunately these tubes do not allow for an extended storage of samples before processing and prevent EDTA tubes from being shipped at ambient temperature. The aim of our study was to compare the quantity and quality of plasma DNA isolated from EDTA and PAXgene® Blood ccfDNA Tubes in different downstream applications. We enrolled 29 patients in our study. Blood samples were drawn into EDTA and PAXgene® Blood ccfDNA Tubes and were processed on day 0 and day 7 after storage at ambient temperature. The plasma DNA from 10 patients was isolated manually. For the DNA isolation from the plasma of 19 additional patients we used the automated QIAsymphony system. The total amount DNA from all samples was measured with a quantitative real-time PCR assay. In addition the amount of methylated mSHOX2 plasma DNA was determined. While the 7day storage lead to an increased amount of total DNA in almost all EDTA tubes, this effect was only seen in very few PAXgene® Blood ccfDNA Tubes. The stabilization solution which prevents the lysis of blood cells had no effect on the method for quantification of methylated sequences in these samples. The quantity and quality of plasma DNA from both types of blood draw tubes are comparable. DNA from PAXgene® Blood ccfDNA was successfully used for PCR-based quantification of total amount of cell-free DNA and for methylation analysis as well. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Genistein Enhances or Reduces Glycosaminoglycan Quantity in a Cell Type-Specific Manner.

    PubMed

    Lan, Ying; Li, Xiulian; Liu, Xuebo; Hao, Cui; Song, Ni; Ren, Sumei; Wang, Wei; Feng, Ningchuan; Zhang, Lijuan

    2018-06-27

    Genistein is a natural isoflavone enriched in soybeans. It has beneficial effects for patients with mucopolysaccharidose type III through inhibiting glycosaminoglycan biosynthesis. However, other studies indicate that genistein does not always inhibit glycosaminoglycan biosynthesis. To understand the underlying molecular mechanisms, CHOK1, CHO3.1, CHO3.3, and HCT116 cells were treated with genistein and the monosaccharide compositions and quantity of all glycans from the cell lysate were measured after thorough acid hydrolysis followed by HPLC analysis. In addition, the glycosaminoglycan disaccharide compositions were obtained by stable isotope labeling coupled with LC/MS analysis. Genistein treatment reduced the amount of glycans but increased the amount of glycosaminoglycans in HCT116 cells. In contrast, genistein treatment reduced both glycan and glycosaminoglycan quantities in CHOK1, CHO3.1, and CHO3.3 cells in addition to differential changes in glycosaminoglycan disaccharide compositions. Genistein treatment reduced overall glycan quantity but glycosaminoglycan quantities were either increased or decreased in a cell type-dependent manner. © 2018 The Author(s). Published by S. Karger AG, Basel.

  20. Mast cell curve-response in partial Achilles tendon rupture after 830 nm phototherapy.

    PubMed

    Pinfildi, Carlos E; da Silva, Érika P Rampazo; Folha, Roberta A C; Turchetto, Paola C G; Monteiro, Paola Pkp; Antunes, Arainy; Hochman, Bernardo S

    2014-02-01

    The aim of this study was to quantify mast cells at different time intervals after partial Achilles tendon rupture in rats treated with low-level laser therapy (LLLT). There is a high incidence of lesions and ruptures in the Achilles tendon that can take weeks and even months to heal completely. As the mast cells help in the healing repair phase, and LLLT has favorable effects on this tissue repair process, study of this modality on the quantity of mastocytes in the ruptured tendon is relevant. Sixty Wistar rats were subjected to partial Achilles' tendon rupture by direct trauma, randomized into 10 groups, and then divided into the group treated with 80 mW aluminum gallium arsenide infrared laser diode, continuous wave, 2.8 W/cm(2) power density, 40 J/cm(2) energy density, and 1.12 J total energy, and the simulation group. Both the groups were subdivided according to the histological assessment period of the sample, either 6 h, 12 h, 24 h, 2 days, or 3 days after the rupture, to quantify the mastocytes in the Achilles' tendon. The group subjected to LLLT presented a greater quantity of mastocytes in the periods of 6 h, 12 h, 24 h, 2 days, and 3 days after rupture, compared with the simulation groups, but differences were detected between the sample assessment periods only in the simulation group. LLLT was shown to increase the quantity of mastocytes in the assessment periods compared with the simulation groups.

  1. Accuracy of Answers to Cell Lineage Questions Depends on Single-Cell Genomics Data Quality and Quantity.

    PubMed

    Spiro, Adam; Shapiro, Ehud

    2016-06-01

    Advances in single-cell (SC) genomics enable commensurate improvements in methods for uncovering lineage relations among individual cells, as determined by phylogenetic analysis of the somatic mutations harbored by each cell. Theoretically, complete and accurate knowledge of the genome of each cell of an individual can produce an extremely accurate cell lineage tree of that individual. However, the reality of SC genomics is that such complete and accurate knowledge would be wanting, in quality and in quantity, for the foreseeable future. In this paper we offer a framework for systematically exploring the feasibility of answering cell lineage questions based on SC somatic mutational analysis, as a function of SC genomics data quality and quantity. We take into consideration the current limitations of SC genomics in terms of mutation data quality, most notably amplification bias and allele dropouts (ADO), as well as cost, which puts practical limits on mutation data quantity obtained from each cell as well as on cell sample density. We do so by generating in silico cell lineage trees using a dedicated formal language, eSTG, and show how the ability to answer correctly a cell lineage question depends on the quality and quantity of the SC mutation data. The presented framework can serve as a baseline for the potential of current SC genomics to unravel cell lineage dynamics, as well as the potential contributions of future advancement, both biochemical and computational, for the task.

  2. Evaluation and verification of epitaxial process sequence for silicon solar-cell production

    NASA Technical Reports Server (NTRS)

    Redfield, D.

    1981-01-01

    To achieve the program goals, 28 minimodules were fabricated and tested, using 600 cells made from three-inch-diameter wafers processed by the sequence chosen for this purpose. Of these 600 cells, half were made from epitaxially grown layers on potentially low-cost substrates. The other half were made from commercial semiconductor-grade (SG), single-crystal silicon wafers that served as controls. Cell processing was normally performed on mixed lots containing significant numbers of each of these two types of wafers. After evaluation of the performance of all cells, they were separated by types for incorporation into modules that were to be tested for electrical performance and response to environmental stress. A simplified flow chart displaying this scheme, for quantities representing half of the planned total to be processed, is presented.

  3. [Relativity among starch quantity, polysaccharides content and total alkaloid content of Dendrobium loddigesii].

    PubMed

    Zhu, Hua; Teng, Jianbei; Cai, Yi; Liang, Jie; Zhu, Yilin; Wei, Tao

    2011-12-01

    To find out the relativity among starch quantity, polysaccharides content and total alkaloid content of Dendrobium loddigesii. Microscopy-counting process was applied to starch quantity statistics, sulfuric acid-anthrone colorimetry was used to assay polysaccharides content and bromocresol green colorimetry was used to assay alkaloid content. Pearson product moment correlation analysis, Kendall's rank correlation analysis and Spearman's concordance coefficient analysis were applied to study their relativity. Extremely significant positive correlation was found between starch quantity and polysaccharides content, and significant negative correlation between alkaloid content and starch quantity was discovered, as well was between alkaloid content and polysaccharides content.

  4. Fourier Transform Infrared Imaging analysis of dental pulp inflammatory diseases.

    PubMed

    Giorgini, E; Sabbatini, S; Conti, C; Rubini, C; Rocchetti, R; Fioroni, M; Memè, L; Orilisi, G

    2017-05-01

    Fourier Transform Infrared microspectroscopy let characterize the macromolecular composition and distribution of tissues and cells, by studying the interaction between infrared radiation and matter. Therefore, we hypothesize to exploit this analytical tool in the analysis of inflamed pulps, to detect the different biochemical features related to various degrees of inflammation. IR maps of 13 irreversible and 12 hyperplastic pulpitis, together with 10 normal pulps, were acquired, compared with histological findings and submitted to multivariate (HCA, PCA, SIMCA) and statistical (one-way ANOVA) analysis. The fit of convoluted bands let calculate meaningful band area ratios (means ± s.d., P < 0.05). The infrared imaging analysis pin-pointed higher amounts of water and lower quantities of type I collagen in all inflamed pulps. Specific vibrational markers were defined for irreversible pulpitis (Lipids/Total Biomass, PhII/Total Biomass, CH 2 /CH 3 , and Ty/AII) and hyperplastic ones (OH/Total Biomass, Collagen/Total Biomass, and CH 3 Collagen/Total Biomass). The study confirmed that FTIR microspectroscopy let discriminate tissues' biological features. The infrared imaging analysis evidenced, in inflamed pulps, alterations in tissues' structure and composition. Changes in lipid metabolism, increasing amounts of tyrosine, and the occurrence of phosphorylative processes were highlighted in irreversible pulpitis, while high amounts of water and low quantities of type I collagen were detected in hyperplastic samples. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  5. Enhanced oral bioavailability of valsartan using a polymer-based supersaturable self-microemulsifying drug delivery system.

    PubMed

    Yeom, Dong Woo; Chae, Bo Ram; Son, Ho Yong; Kim, Jin Han; Chae, Jun Soo; Song, Seh Hyon; Oh, Dongho; Choi, Young Wook

    2017-01-01

    A novel, supersaturable self-microemulsifying drug delivery system (S-SMEDDS) was successfully formulated to enhance the dissolution and oral absorption of valsartan (VST), a poorly water-soluble drug, while reducing the total quantity for administration. Poloxamer 407 is a selectable, supersaturating agent for VST-containing SMEDDS composed of 10% Capmul ® MCM, 45% Tween ® 20, and 45% Transcutol ® P. The amounts of SMEDDS and Poloxamer 407 were chosen as formulation variables for a 3-level factorial design. Further optimization was established by weighting different levels of importance on response variables for dissolution and total quantity, resulting in an optimal S-SMEDDS in large quantity (S-SMEDDS_LQ; 352 mg in total) and S-SMEDDS in reduced quantity (S-SMEDDS_RQ; 144.6 mg in total). Good agreement was observed between predicted and experimental values for response variables. Consequently, compared with VST powder or suspension and SMEDDS, both S-SMEDDS_LQ and S-SMEDDS_RQ showed excellent in vitro dissolution and in vivo oral bioavailability in rats. The magnitude of dissolution and absorption-enhancing capacities using quantity-based comparisons was in the order S-SMEDDS_RQ > S-SMEDDS_LQ > SMEDDS > VST powder or suspension. Thus, we concluded that, in terms of developing an effective SMEDDS preparation with minimal total quantity, S-SMEDDS_RQ is a promising candidate.

  6. Secretory vesicles of immune cells contain only a limited number of interleukin 6 molecules.

    PubMed

    Verboogen, Daniëlle R J; Ter Beest, Martin; Honigmann, Alf; van den Bogaart, Geert

    2018-05-01

    Immune cells communicate by releasing large quantities of cytokines. Although the mechanisms of cytokine secretion are increasingly understood, quantitative knowledge of the number of cytokines per vesicle is still lacking. Here, we measured with quantitative microscopy the release rate of vesicles potentially carrying interleukin-6 (IL-6) in human dendritic cells. By comparing this to the total secreted IL-6, we estimate that secretory vesicles contain about 0.5-3 IL-6 molecules, but with a large spread among cells/donors. Moreover, IL-6 did not accumulate within most cells, indicating that synthesis and not trafficking is the bottleneck for IL-6 production. IL-6 accumulated in the Golgi apparatus only in ~ 10% of the cells. Understanding how immune cells produce cytokines is important for designing new immunomodulatory drugs. © 2018 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.

  7. Relative quantity judgments in South American sea lions (Otaria flavescens).

    PubMed

    Abramson, José Z; Hernández-Lloreda, Victoria; Call, Josep; Colmenares, Fernando

    2011-09-01

    There is accumulating evidence that a variety of species possess quantitative abilities although their cognitive substrate is still unclear. This study is the first to investigate whether sea lions (Otaria flavescens), in the absence of training, are able to assess and select the larger of two sets of quantities. In Experiment 1, the two sets of quantities were presented simultaneously as whole sets, that is, the subjects could compare them directly. In Experiment 2, the two sets of quantities were presented item-by-item, and the totality of items was never visually available at the time of choice. For each type of presentation, we analysed the effect of the ratio between quantities, the difference between quantities and the total number of items presented. The results showed that (1) sea lions can make relative quantity judgments successfully and (2) there is a predominant influence of the ratio between quantities on the subjects' performance. The latter supports the idea that an analogue representational mechanism is responsible for sea lions' relative quantities judgments. These findings are consistent with previous reports of relative quantities judgments in other species such as monkeys and apes and suggest that sea lions might share a similar mechanism to compare and represent quantities.

  8. Peloruside B, a Potent Antitumor Macrolide from the New Zealand Marine Sponge Mycale hentscheli: Isolation, Structure, Total Synthesis and Bioactivity

    PubMed Central

    Singh, A. Jonathan; Xu, Chun-Xiao; Xu, Xiaoming; West, Lyndon M.; Wilmes, Anja; Chan, Ariane; Hamel, Ernest; Miller, John H.; Northcote, Peter T.; Ghosh, Arun K.

    2009-01-01

    Peloruside B (2), a natural congener of peloruside A (1), was isolated in sub-milligram quantities from the New Zealand marine sponge Mycale hentscheli. Peloruside B promotes microtubule polymerization and arrests cells in the G2M phase of mitosis similar to paclitaxel, and its bioactivity was comparable to that of peloruside A. NMR-directed isolation, structure elucidation, structure confirmation by total synthesis and bioactivity of peloruside B are described in this article. The synthesis features Sharpless dihydroxylation, Brown's asymmetric allylboration reaction, reductive aldol coupling, Yamaguchi macrolactonization and selective methylation. PMID:19957922

  9. Quality‐Quantity Control Culture Enhances Vasculogenesis and Wound Healing Efficacy of Human Diabetic Peripheral Blood CD34+ Cells

    PubMed Central

    Masuda, Haruchika; Fujimura, Satoshi; Ito‐Hirano, Rie; Arita, Kayo; Kakinuma, Yusuke; Hagiwara, Hiroko; Kado, Makiko; Hayashi, Ayato; Mita, Tomoya; Ogawa, Takasuke; Watada, Hirotaka; Mizuno, Hiroshi; Sawada, Naoki; Asahara, Takayuki

    2018-01-01

    Abstract Autologous endothelial progenitor cell (EPC) therapy is commonly used to stimulate angiogenesis in ischemic repair and wound healing. However, low total numbers and functional deficits of EPCs make autologous EPC therapy ineffective in diabetes. Currently, no known ex vivo culture techniques can expand and/or ameliorate the functional deficits of EPCs for clinical usage. Recently, we showed that a quality‐quantity culture (QQc) system restores the vasculogenic and wound‐healing efficacy of murine diabetic EPCs. To validate these results and elucidate the mechanism in a translational study, we evaluated the efficacy of this QQc system to restore the vasculogenic potential of diabetic human peripheral blood (PB) CD34+ cells. CD34+ cells purified from PB of diabetic and healthy patients were subjected to QQc. Gene expression, vascular regeneration, and expression of cytokines and paracrine mediators were analyzed. Pre‐ or post‐QQc diabetic human PB‐CD34+ cells were transplanted into wounded BALB/c nude mice and streptozotocin‐induced diabetic mice to assess functional efficacy. Post‐QQc diabetic human PB‐CD34+ cell therapy significantly accelerated wound closure, re‐epithelialization, and angiogenesis. The higher therapeutic efficacy of post‐QQc diabetic human PB‐CD34+ cells was attributed to increased differentiation ability of diabetic CD34+ cells, direct vasculogenesis, and enhanced expression of angiogenic factors and wound‐healing genes. Thus, QQc can significantly enhance the therapeutic efficacy of human PB‐CD34+ cells in diabetic wounds, overcoming the inherent limitation of autologous cell therapy in diabetic patients, and could be useful for treatment of not only wounds but also other ischemic diseases. Stem Cells Translational Medicine 2018;7:428–438 PMID:29573563

  10. 7 CFR 929.105 - Reporting.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE CRANBERRIES GROWN IN STATES OF... succeeding fiscal period showing: (1) The total quantity of cranberries the handler acquired and the total quantity of cranberries and Vaccinium oxycoccus cranberries the handler handled from the beginning of the...

  11. 7 CFR 929.105 - Reporting.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE CRANBERRIES GROWN IN STATES OF... succeeding fiscal period showing: (1) The total quantity of cranberries the handler acquired and the total quantity of cranberries and Vaccinium oxycoccus cranberries the handler handled from the beginning of the...

  12. 7 CFR 929.105 - Reporting.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... AND ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE CRANBERRIES GROWN IN STATES OF... succeeding fiscal period showing: (1) The total quantity of cranberries the handler acquired and the total quantity of cranberries and Vaccinium oxycoccus cranberries the handler handled from the beginning of the...

  13. 7 CFR 929.105 - Reporting.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... AND ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE CRANBERRIES GROWN IN STATES OF... succeeding fiscal period showing: (1) The total quantity of cranberries the handler acquired and the total quantity of cranberries and Vaccinium oxycoccus cranberries the handler handled from the beginning of the...

  14. 7 CFR 1207.533 - Availability of records.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... AGREEMENTS AND ORDERS; MISCELLANEOUS COMMODITIES), DEPARTMENT OF AGRICULTURE POTATO RESEARCH AND PROMOTION... records on the total quantities of potatoes imported and on the total quantities of potato products imported, and a record of each importation of potatoes, potato products, and seed potatoes including...

  15. 7 CFR 1207.533 - Availability of records.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... AGREEMENTS AND ORDERS; MISCELLANEOUS COMMODITIES), DEPARTMENT OF AGRICULTURE POTATO RESEARCH AND PROMOTION... records on the total quantities of potatoes imported and on the total quantities of potato products imported, and a record of each importation of potatoes, potato products, and seed potatoes including...

  16. Potential radiological impact of tornadoes on the safety of Nuclear Fuel Services' West Valley Fuel Reprocessing Plant. 2. Reentrainment and discharge of radioactive materials

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Davis, W Jr

    1981-07-01

    This report describes results of a parametric study of quantities of radioactive materials that might be discharged by a tornado-generated depressurization on contaminated process cells within the presently inoperative Nuclear Fuel Services' (NFS) fuel reprocessing facility near West Valley, New York. The study involved the following tasks: determining approximate quantities of radioactive materials in the cells and characterizing particle-size distribution; estimating the degree of mass reentrainment from particle-size distribution and from air speed data presented in Part 1; and estimating the quantities of radioactive material (source term) released from the cells to the atmosphere. The study has shown that improperlymore » sealed manipulator ports in the Process Mechanical Cell (PMC) present the most likely pathway for release of substantial quantities of radioactive material in the atmosphere under tornado accident conditions at the facility.« less

  17. 14 CFR 31.85 - Required basic equipment.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... indicator. (b) For hot air balloons: (1) A fuel quantity gauge. If fuel cells are used, means must be incorporated to indicate to the crew the quantity of fuel in each cell during flight. The means must be calibrated in appropriate units or in percent of fuel cell capacity. (2) An envelope temperature indicator...

  18. 14 CFR 31.85 - Required basic equipment.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... indicator. (b) For hot air balloons: (1) A fuel quantity gauge. If fuel cells are used, means must be incorporated to indicate to the crew the quantity of fuel in each cell during flight. The means must be calibrated in appropriate units or in percent of fuel cell capacity. (2) An envelope temperature indicator...

  19. 14 CFR 31.85 - Required basic equipment.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... indicator. (b) For hot air balloons: (1) A fuel quantity gauge. If fuel cells are used, means must be incorporated to indicate to the crew the quantity of fuel in each cell during flight. The means must be calibrated in appropriate units or in percent of fuel cell capacity. (2) An envelope temperature indicator...

  20. 14 CFR 31.85 - Required basic equipment.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... indicator. (b) For hot air balloons: (1) A fuel quantity gauge. If fuel cells are used, means must be incorporated to indicate to the crew the quantity of fuel in each cell during flight. The means must be calibrated in appropriate units or in percent of fuel cell capacity. (2) An envelope temperature indicator...

  1. Large-scale generation of cell-derived nanovesicles

    NASA Astrophysics Data System (ADS)

    Jo, W.; Kim, J.; Yoon, J.; Jeong, D.; Cho, S.; Jeong, H.; Yoon, Y. J.; Kim, S. C.; Gho, Y. S.; Park, J.

    2014-09-01

    Exosomes are enclosed compartments that are released from cells and that can transport biological contents for the purpose of intercellular communications. Research into exosomes is hindered by their rarity. In this article, we introduce a device that uses centrifugal force and a filter with micro-sized pores to generate a large quantity of cell-derived nanovesicles. The device has a simple polycarbonate structure to hold the filter, and operates in a common centrifuge. Nanovesicles are similar in size and membrane structure to exosomes. Nanovesicles contain intracellular RNAs ranging from microRNA to mRNA, intracellular proteins, and plasma membrane proteins. The quantity of nanovesicles produced using the device is 250 times the quantity of naturally secreted exosomes. Also, the quantity of intracellular contents in nanovesicles is twice that in exosomes. Nanovesicles generated from murine embryonic stem cells can transfer RNAs to target cells. Therefore, this novel device and the nanovesicles that it generates are expected to be used in exosome-related research, and can be applied in various applications such as drug delivery and cell-based therapy.

  2. Isolation and purification of total RNA from Streptococcus mutans in suspension cultures and biofilms.

    PubMed

    Cury, Jaime Aparecido; Seils, Jennifer; Koo, Hyun

    2008-01-01

    The presence of extracellular polysaccharides matrix makes extraction and purification of RNA from Streptococcus mutans within biofilms challenging. In this study, several approaches to purify RNA extracted from S. mutans in suspension cultures and biofilms were examined. The combination of sonication (3 pulses of 30 s at 7 W), suspension in NAES buffer (50 mM sodium acetate buffer, 10 mM EDTA and 1% SDS; pH 5.0) and homogenization-mechanical cells disruption in NAES- acid phenol:chloroform, yielded 9.04 mg (or 0.52 mg) of crude preparation of RNA per 100 mg of total cell (or biofilm) dry-weight. The crude RNA preparations were subjected to various DNAse I treatments. The combination of DNAse I in silica-gel based column followed by recombinant DNase I in solution provided the best genomic DNA removal, resulting in 4.35 mg (or 0.06 mg) of purified RNA per 100 mg of total cell (or biofilm) dry-weight. The cDNAs generated from the purified RNA sample were efficiently amplified using gtfB S. mutans-specific primers. The results showed a method that yields high-quality RNA from both planktonic cells and biofilms of S. mutans in sufficient quantity and quality for real-time RT-PCR analyses.

  3. 7 CFR 966.40 - Expenses.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE TOMATOES GROWN IN FLORIDA Order... share of such expense shall be proportionate to the ratio between the total quantity of tomatoes handled by him as the first handler thereof during a fiscal period and the total quantity of tomatoes handled...

  4. Partitioning of Electromagnetic Energy Inputs to the Thermosphere during Geomagnetic Disturbances

    DTIC Science & Technology

    2012-06-01

    boundary of a local flux tube volume is an equipotential . Figure 4 contains maps of Poynting flux normal to a 500 km altitude surface and maps of height...as a cell quantity throughout its computational volume, we are able to generate maps of the Poynting flux, ⃗ ⃗⃗⃗⃗⃗⃗ , on altitude surfaces at...the top of the thermosphere. We used separate modules to integrate the Poynting flux over this surface to compute the total electromagnetic energy

  5. Osteoblast response to porous titanium and biomimetic surface: In vitro analysis.

    PubMed

    Prado, Renata Falchete do; de Oliveira, Fernanda Saraiva; Nascimento, Rodrigo Dias; de Vasconcellos, Luana Marotta Reis; Carvalho, Yasmin Rodarte; Cairo, Carlos Alberto Alves

    2015-01-01

    This study analyzed the behavior of human osteoblasts cultured on porous titanium specimens, with and without biomimetic treatment, compared to dense titanium. The experiment had seven groups: Group 1: cells cultured on polystyrene of culture plate wells; Group 2: cells cultured on dense titanium specimen; Group 3: specimen with 33.79% of pores; Group 4: 41.79% of pores; Groups 5, 6 and 7: specimens similar to groups 2, 3 and 4, yet with biomimetic treatment. Real time-polymerase chain reaction with reverse transcription of the following genes was performed: prostaglandin E2 synthase, integrin β1, osterix, Runx2, Interleukin 6, macrophage colony stimulating factor, apolipoprotein E and others. The study achieved data on cell adhesion, growth and viability, total protein content, alkaline phosphatase activity and quantity of mineralized nodule formations. Data were statistically evaluated. Adherent cells and alkaline phosphatase activity were similar in titanium specimens, regardless of the groups. Biomimetic treatment reduced the total protein activity and the viability of tested cells. Most tested genes had statistically similar expression in all groups. The tested porosities did not cause alterations in osteoblast behavior and the biomimetic treatment impaired the biocompatibility of titanium causing cytotoxicity. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Trans-inner Cell Mass Injection of Embryonic Stem Cells Leads to Higher Chimerism Rates.

    PubMed

    Scott, Gregory J; Gruzdev, Artiom; Hagler, Thomas B; Ray, Manas K

    2018-05-29

    In an effort to increase efficiency in the creation of genetically modified mice via ES Cell methodologies, we present an adaptation to the current blastocyst injection protocol. Here we report that a simple rotation of the embryo, and injection through Trans-Inner cell mass (TICM) increased the percentage of chimeric mice from 31% to 50%, with no additional equipment or further specialized training. 26 different inbred clones, and 35 total clones were injected over a period of 9 months. There was no significant difference in either pregnancy rate or recovery rate of embryos between traditional injection techniques and TICM. Therefore, without any major alteration in the injection process and a simple positioning of the blastocyst and injecting through the ICM, releasing the ES cells into the blastocoel cavity can potentially improve the quantity of chimeric production and subsequent germline transmission.

  7. 7 CFR 906.151 - Reports.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 8 2013-01-01 2013-01-01 false Reports. 906.151 Section 906.151 Agriculture... RIO GRANDE VALLEY IN TEXAS Rules and Regulations § 906.151 Reports. (a) During each fiscal period... the request an accurate report showing the total quantity or oranges and the total quantity of...

  8. 7 CFR 906.151 - Reports.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 8 2014-01-01 2014-01-01 false Reports. 906.151 Section 906.151 Agriculture... RIO GRANDE VALLEY IN TEXAS Rules and Regulations § 906.151 Reports. (a) During each fiscal period... the request an accurate report showing the total quantity or oranges and the total quantity of...

  9. Environmental Assessment for the Space Based Infrared System (SBIRS) Mission Control Station for Defense Support Program Consolidation

    DTIC Science & Technology

    1996-04-01

    use of products containing hazardous materials (e.g., cleaning products , spray and liquid solvents, toner cartridges, etc.). The total quantity of...products containing hazardous materials (e.g., cleaning products , spray and liquid solvents, toner cartridges, etc.). The total quantity of hazardous

  10. Practical thermodynamic quantities for aqueous vanadium- and iron-based flow batteries

    DOE PAGES

    Hudak, Nicholas S.

    2013-12-31

    A simple method for experimentally determining thermodynamic quantities for flow battery cell reactions is presented. Equilibrium cell potentials, temperature derivatives of cell potential (d E/d T), Gibbs free energies, and entropies are reported here for all-vanadium, iron–vanadium, and iron–chromium flow cells with state-of-the-art solution compositions. Proof is given that formal potentials and formal temperature coefficients can be used with modified forms of the Nernst Equation to quantify the thermodynamics of flow cell reactions as a function of state-of-charge. Such empirical quantities can be used in thermo-electrochemical models of flow batteries at the cell or system level. In most cases, themore » thermodynamic quantities measured here are significantly different from standard values reported and used previously in the literature. The data reported here are also useful in the selection of operating temperatures for flow battery systems. Because higher temperatures correspond to lower equilibrium cell potentials for the battery chemistries studied here, it can be beneficial to charge a cell at higher temperature and discharge at lower temperature. As a result, proof-of-concept of improved voltage efficiency with the use of such non-isothermal cycling is given for the all-vanadium redox flow battery, and the effect is shown to be more pronounced at lower current densities.« less

  11. Simplified Bioreactor For Growing Mammalian Cells

    NASA Technical Reports Server (NTRS)

    Spaulding, Glenn F.

    1995-01-01

    Improved bioreactor for growing mammalian cell cultures developed. Designed to support growth of dense volumes of mammalian cells by providing ample, well-distributed flows of nutrient solution with minimal turbulence. Cells relatively delicate and, unlike bacteria, cannot withstand shear forces present in turbulent flows. Bioreactor vessel readily made in larger sizes to accommodate greater cell production quantities. Molding equipment presently used makes cylinders up to 30 centimeters long. Alternative sintered plastic techniques used to vary pore size and quantity, as necessary.

  12. Separation of granulocytes from whole blood by leukoadhesion, phase 1

    NASA Technical Reports Server (NTRS)

    1976-01-01

    Capillary glass tubes are investigated for the separation and retrieval of large quantities of viable granulocytes and monocytes from whole blood on a continuous basis from a single donor. This effort represented the feasibility demonstration of a three phase program for development of a capillary tube cell separation device. The activity included the analysis and parametric laboratory testing with subscale models required to design a prototype device. Capillary tubes 40 cm long with a nominal 0.030 cm internal diameter yielded the highest total process efficiency. Recovery efficiencies as high as 89% of the adhering cell population were obtained. Granulocyte phagocytosis of latex particles indicated approximately 90% viability. Monocytes recovered from the separation column retained their capability to stimulate human bone marrow colony growth, as demonstrated in an in vitro cell culture assay.

  13. Human Bone Marrow-Derived Mesenchymal Stem Cells Display Enhanced Clonogenicity but Impaired Differentiation With Hypoxic Preconditioning

    PubMed Central

    Boyette, Lisa B.; Creasey, Olivia A.; Guzik, Lynda; Lozito, Thomas

    2014-01-01

    Stem cells are promising candidate cells for regenerative applications because they possess high proliferative capacity and the potential to differentiate into other cell types. Mesenchymal stem cells (MSCs) are easily sourced but do not retain their proliferative and multilineage differentiative capabilities after prolonged ex vivo propagation. We investigated the use of hypoxia as a preconditioning agent and in differentiating cultures to enhance MSC function. Culture in 5% ambient O2 consistently enhanced clonogenic potential of primary MSCs from all donors tested. We determined that enhanced clonogenicity was attributable to increased proliferation, increased vascular endothelial growth factor secretion, and increased matrix turnover. Hypoxia did not impact the incidence of cell death. Application of hypoxia to osteogenic cultures resulted in enhanced total mineral deposition, although this effect was detected only in MSCs preconditioned in normoxic conditions. Osteogenesis-associated genes were upregulated in hypoxia, and alkaline phosphatase activity was enhanced. Adipogenic differentiation was inhibited by exposure to hypoxia during differentiation. Chondrogenesis in three-dimensional pellet cultures was inhibited by preconditioning with hypoxia. However, in cultures expanded under normoxia, hypoxia applied during subsequent pellet culture enhanced chondrogenesis. Whereas hypoxic preconditioning appears to be an excellent way to expand a highly clonogenic progenitor pool, our findings suggest that it may blunt the differentiation potential of MSCs, compromising their utility for regenerative tissue engineering. Exposure to hypoxia during differentiation (post-normoxic expansion), however, appears to result in a greater quantity of functional osteoblasts and chondrocytes and ultimately a larger quantity of high-quality differentiated tissue. PMID:24436440

  14. Steady-state configurations and dynamics of the MreB helix within bacteria

    NASA Astrophysics Data System (ADS)

    Rutenberg, Andrew; Allard, Jun

    2007-03-01

    We present a quantitative model of the actin-like MreB cytoskeleton that is present in many prokaryotes. Individual MreB polymers are bundled into a supra-molecular array to make up helical cables. The cell wall imposes constraint forces through a global elasticity model. With variational techniques and stochastic simulations we obtain relationships between observable quantities such as the pitch of the helix, the total abundance of MreB molecules, and the thickness of the MreB cables. We address changes expected with slow cell growth, as well as turnover dynamics that are relevant to FRAP studies. We also address polarized macromolecular trafficking along the MreB cables without motor proteins.

  15. Circulating miRNAs as Potential Alternative Cell Signaling Associated with Maternal Recognition of Pregnancy in the Mare.

    PubMed

    Klohonatz, Kristin M; Cameron, Ashely D; Hergenreder, Joanna R; da Silveira, Juliano C; Belk, Aeriel D; Veeramachaneni, D N R; Bouma, Gerrit J; Bruemmer, Jason E

    2016-12-01

    During early pregnancy, the conceptus and mare communicate to establish pregnancy. Cell-secreted vesicles (e.g., exosomes) have been reported in serum. Exosomes contain bioactive materials, such as miRNA, that can mediate cell responses. We hypothesized that a) exosomes are present in mare circulation and quantity varies with pregnancy status, b) exosomes contain miRNAs unique to pregnancy status, and c) miRNAs target pathways in endometrium based upon pregnancy status of the mare. First, serum samples were obtained from mares in a crossover design, with each mare providing samples from a pregnant and nonmated control cycle (n = 3/sample day) on Days 12, 14, 16, and 18 postovulation. Flow cytometry revealed the presence of serum microvesicles in mares in two different-sized populations (greater than or less than 100 nm), validated by transmission electron microscopy. Second, serum was collected on Days 9, 11, and 13 (n = 4/day), and endometrial biopsies were collected on Days 11 and 13 (n = 3/day) from pregnant and nonmated mares. Total RNA from serum exosomes was evaluated with quantitative RT-PCR using equine-specific miRNA sequences. A total of 12 miRNAs were found in different quantities on the specified days. Pathway analysis suggested that miRNAs targeted focal adhesion molecules (FAMs). Transcripts corresponding to FAMs were evaluated in endometrial biopsies. Protein levels and localization for PAK6 and RAF1 were further evaluated. Our data suggest that serum exosomes contain miRNA that differ based upon pregnancy status, and may affect mRNA expression related to focal adhesion pathway in the endometrium, with a potential role in maternal recognition of pregnancy. © 2016 by the Society for the Study of Reproduction, Inc.

  16. Meta-analysis to estimate the load of Leptospira excreted in urine: beyond rats as important sources of transmission in low-income rural communities.

    PubMed

    Barragan, Veronica; Nieto, Nathan; Keim, Paul; Pearson, Talima

    2017-01-28

    Leptospirosis is a major zoonotic disease with widespread distribution and a large impact on human health. Carrier animals excrete pathogenic Leptospira primarily in their urine. Infection occurs when the pathogen enters a host through mucosa or small skin abrasions. Humans and other animals are exposed to the pathogen by direct contact with urine, contaminated soil or water. While many factors influence environmental cycling and the transmission of Leptospira to humans, the load of pathogenic Leptospira in the environment is likely to play a major role. Peridomestic rats are often implicated as a potential source of human disease; however exposure to other animals is a risk factor as well. The aim of this report is to highlight the importance of various carrier animals in terms of the quantity of Leptospira shed into the environment. For this, we performed a systematic literature review and a meta-analysis of the amount of pathogen that various animal species shed in their urine. The quantity of pathogen has been reported for cows, deer, dogs, humans, mice, and rats, in a total of 14 research articles. We estimated the average Leptospira per unit volume shed by each animal species, and the daily environmental contribution by considering the total volume of urine excreted by each carrier animal. Rats excrete the highest quantity of Leptospira per millilitre of urine (median = 5.7 × 10 6  cells), but large mammals excrete much more urine and thus shed significantly more Leptospira per day (5.1 × 10 8 to 1.3 × 10 9  cells). Here we illustrate how, in a low-income rural Ecuadorian community, host population demographics, and prevalence of Leptospira infection can be integrated with estimates of shed Leptospira to suggest that peridomestic cattle may be more important than rats in environmental cycling and ultimately, transmission to humans.

  17. Statistical Characterization of 18650-Format Lithium-Ion Cell Thermal Runaway Energy Distributions

    NASA Technical Reports Server (NTRS)

    Walker, William Q.; Rickman, Steven; Darst, John; Finegan, Donal; Bayles, Gary; Darcy, Eric

    2017-01-01

    Effective thermal management systems, designed to handle the impacts of thermal runaway (TR) and to prevent cell-to-cell propagation, are key to safe operation of lithium-ion (Li-ion) battery assemblies. Critical factors for optimizing these systems include the total energy released during a single cell TR event and the fraction of the total energy that is released through the cell casing vs. through the ejecta material. A unique calorimeter was utilized to examine the TR behavior of a statistically significant number of 18650-format Li-ion cells with varying manufacturers, chemistries, and capacities. The calorimeter was designed to contain the TR energy in a format conducive to discerning the fractions of energy released through the cell casing vs. through the ejecta material. Other benefits of this calorimeter included the ability to rapidly test of large quantities of cells and the intentional minimization of secondary combustion effects. High energy (270 Wh/kg) and moderate energy (200 Wh/kg) 18650 cells were tested. Some of the cells had an imbedded short circuit (ISC) device installed to aid in the examination of TR mechanisms under more realistic conditions. Other variations included cells with bottom vent (BV) features and cells with thin casings (0.22 1/4m). After combining the data gathered with the calorimeter, a statistical approach was used to examine the probability of certain TR behavior, and the associated energy distributions, as a function of capacity, venting features, cell casing thickness and temperature.

  18. [Influence of Different Straws Returning with Landfill on Soil Microbial Community Structure Under Dry and Water Farming].

    PubMed

    Lan, Mu-ling; Gao, Ming

    2015-11-01

    Based on rice, wheat, corn straw and rape, broad bean green stalk as the research object, using phospholipid fatty acid (PLFA) method, combining principal component analysis method to study the soil microbial quantity, distribution of flora, community structure characteristics under dry and water farming as two different cultivated land use types. The PLFA analysis results showed that: under dry farming, total PLFA quantity ranged 8.35-25.15 nmol x g(-1), showed rape > broad bean > corn > rice > wheat, rape and broad bean significantly increased total PLFA quantity by 1.18 and 1.08 times compared to the treatment without straw; PLFA quantity of bacterial flora in treatments with straws was higher than that without straw, and fungal biomass was significantly increased, so was the species richness of microbial community. Under water faming, the treatments of different straws returning with landfill have improved the PLFA quantity of total soil microbial and flora comparing with the treatment without straw, fungi significantly increased, and species richness of microbial communities value also increased significantly. Total PLFA quantity ranged 4.04-22.19 nmol x g(-1), showed rice > corn > wheat > broad bean > rape, which in rape and broad bean treatments were lower than the treatment without straw; fungal PLFA amount in 5 kinds of straw except broad bean treatment was significantly higher than that of the treatment without straw, bacteria and total PLFA quantity in broad bean processing were significantly lower than those of other treatments, actinomycetes, G+, G- had no significant difference between all treatments; rice, wheat, corn, rape could significantly increase the soil microbial species richness index and dominance index under water faming. The results of principal component analysis showed that broad bean green stalk had the greatest impact on the microbial community structure in the dry soil, rape green stalk and wheat straw had the biggest influence on soil microbial community structure in water soil.

  19. 77 FR 28259 - Mailings of Lithium Batteries

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-14

    ... containing lithium metal or lithium-ion cells or batteries and applies regardless of quantity, size, watt... ``lithium content'' for secondary lithium-ion batteries when describing maximum quantity limits. In addition...-ion (Rechargeable) Cells and Batteries [Revise 10.20.6 as follows:] Small consumer-type lithium-ion...

  20. Thermodynamic properties of water in confined environments: a Monte Carlo study

    NASA Astrophysics Data System (ADS)

    Gladovic, Martin; Bren, Urban; Urbic, Tomaž

    2018-05-01

    Monte Carlo simulations of Mercedes-Benz water in a crowded environment were performed. The simulated systems are representative of both composite, porous or sintered materials and living cells with typical matrix packings. We studied the influence of overall temperature as well as the density and size of matrix particles on water density, particle distributions, hydrogen bond formation and thermodynamic quantities. Interestingly, temperature and space occupancy of matrix exhibit a similar effect on water properties following the competition between the kinetic and the potential energy of the system, whereby temperature increases the kinetic and matrix packing decreases the potential contribution. A novel thermodynamic decomposition approach was applied to gain insight into individual contributions of different types of inter-particle interactions. This decomposition proved to be useful and in good agreement with the total thermodynamic quantities especially at higher temperatures and matrix packings, where higher-order potential-energy mixing terms lose their importance.

  1. Secondary metabolites and metal content dynamics in Teucrium montanum L. and Teucrium chamaedrys L. from habitats with serpentine and calcareous substrate.

    PubMed

    Zlatić, Nenad M; Stanković, Milan S; Simić, Zoran S

    2017-03-01

    The purpose of this comparative analysis is the determination of the total quantity of metals (Mg, Ca, K, Ni, Fe, Mn, Zn, Cu, Cr and Pb) in soil samples, above ground plant parts and tea made of plants Teucrium montanum and T. chamaedrys from different serpentine and calcareous habitats as well as of the total quantity of phenolic compounds and antioxidant activity. The obtained results showed that the quantities of certain metals (Mg, Fe, Ni and Mn) in the soil from the serpentine habitats were greater in comparison with other metals (Ca, Zn and Pb) which were more frequently found in the soil from the calcareous habitats. The results demonstrated that the analysed plant samples from the serpentine habitats contained higher quantity of Fe, Ni and Cr as opposed to the plant samples from the calcareous habitats which contained greater quantity of Ca and Zn. Although the studied species accumulate analysed metals in different quantities, depending on the substrate type, they are not hyperaccumulators of these metals. The use of these species from serpentine habitats for tea preparation is safe to a great extent, because in spite of the determined metal absorption by plant organs, the tea does not contain dangerous quantity of heavy metals. The results showed greater total quantity of phenolic compounds and the higher level of antioxidant activity in the plant samples from serpentine habitats in comparison with the samples from calcareous habitats, which is an indicator of one of the mechanisms of adaptation to the serpentine habitat conditions.

  2. 40 CFR 355.14 - Do I have to aggregate extremely hazardous substances to determine the total quantity present?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 28 2011-07-01 2011-07-01 false Do I have to aggregate extremely hazardous substances to determine the total quantity present? 355.14 Section 355.14 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SUPERFUND, EMERGENCY PLANNING, AND COMMUNITY RIGHT-TO...

  3. Testing of concrete by laser ablation

    DOEpatents

    Flesher, Dann J.; Becker, David L.; Beem, William L.; Berry, Tommy C.; Cannon, N. Scott

    1997-01-01

    A method of testing concrete in a structure in situ, by: directing a succession of pulses of laser radiation at a point on the structure so that each pulse effects removal of a quantity of concrete and transfers energy to the concrete; detecting a characteristic of energy which has been transferred to the concrete; determining, separately from the detecting step, the total quantity of concrete removed by the succession of pulses; and calculating a property of the concrete on the basis of the detected energy characteristic and the determined total quantity of concrete removed.

  4. 40 CFR 98.386 - Data reporting requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... (CONTINUED) MANDATORY GREENHOUSE GAS REPORTING Suppliers of Coal-based Liquid Fuels § 98.386 Data reporting... measurement standard method or other industry standard practice used. For natural gas liquids, quantity shall... site, report the total annual quantity in metric tons or barrels. For natural gas liquids, quantity...

  5. 40 CFR 98.386 - Data reporting requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... (CONTINUED) MANDATORY GREENHOUSE GAS REPORTING Suppliers of Coal-based Liquid Fuels § 98.386 Data reporting... measurement standard method or other industry standard practice used. For natural gas liquids, quantity shall... site, report the total annual quantity in metric tons or barrels. For natural gas liquids, quantity...

  6. 40 CFR 98.386 - Data reporting requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... (CONTINUED) MANDATORY GREENHOUSE GAS REPORTING Suppliers of Coal-based Liquid Fuels § 98.386 Data reporting... measurement standard method or other industry standard practice used. For natural gas liquids, quantity shall... site, report the total annual quantity in metric tons or barrels. For natural gas liquids, quantity...

  7. 40 CFR 98.386 - Data reporting requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... (CONTINUED) MANDATORY GREENHOUSE GAS REPORTING Suppliers of Coal-based Liquid Fuels § 98.386 Data reporting... measurement standard method or other industry standard practice used. For natural gas liquids, quantity shall... site, report the total annual quantity in metric tons or barrels. For natural gas liquids, quantity...

  8. First total synthesis of a novel amide alkaloid derived from Aconitum taipeicum and its anticancer activity.

    PubMed

    Zhang, Xinxin; Li, Dandan; Xue, Xuanji; Zhang, Yan; Zhang, Jie; Huang, Chen; Guo, Zengjun; Tadesse, Nigatu

    2018-01-01

    A concise total synthesis of a naturally occurring 3-isopropyl-tetrahydropyrrolo[1, 2-a]pyrimidine-2, 4(1H, 3H)-dione (ITPD) isolated from Aconitum taipeicum with a three-step approach was depicted in this study for the first time. Two key intermediates, diethyl isopropylmalonate (2) and pyrrolidin-2-amine (3), being synthsesised separately from initial diethyl malonate (4) and 3, 4-dihydro-2H-pyrrol-5-amine (5), were utilised to obtain the compound entitled ITPD. ITPD showed a promising anticancer activity in vitro on SMMC-7721 cell lines. Flow cytometry and cell cycle analysis revealed that ITPD could induce apoptosis and cell cycle arrest in S phase. The occurrence of apoptosis possibly attributed to the mechanism that ITPD could mediate the mitochondrial pathway through activating caspase-3/9 and increasing the ratio of Bax/Bcl-2 to finally trigger cell apoptosis and DNA damage. Collectively, the possibility to produce sufficient quantity of synthetic ITPD provided the base for further bio-evaluation in vivo and in vitro. The bioactive assay suggested that it may be a potential candidate for further chemical optimisation and use in cancer therapy.

  9. The differentiation directions of the bone marrow stromal cells under modeling microgravity

    NASA Astrophysics Data System (ADS)

    Nesterenko, Olga; Rodionova, Natalia; Katkova, Olena

    Within experiments on rats simulating microgravity by base load remove from back limbs (duration of the experiment 1,5 months) on marrow stromal cells cultures (ex vivo, in vitro) comprising osteogenic cells-predecessors, extracted from femurs, studied their peculiarities of the colony formation ablity, the cell structure, some cytological and ultra-structural characteristics and differentiation direction. It was found that that under microgravity conditions there is a decline of the stromal cells colony formation intensity, decrease of the colonies size and cells mitotic activity that indicates decrease of their growth potential. Both in control and in experiment the colonies were presented by population of low-differentiated cells, differentiated cells and mature cells. The comparative cytological and morphometric analysis have shown that the studied stromal cells in colonies have the smaller sizes, more elongated shape, and higher nucleocytoplasmic ratio. Cells composition in the experiment colonies is reliably different by the ratio of the low-differentiating to being differentiated cells; a ratio of low-differentiated to already differentiated cells; ratio of differentiated cells to total number of all cells. In comparison with control group, amount of the cells passed trough a differentiation stage and mature cells in colonies is decreased by 3 to 4 times. Among the differentiated stromal cells in colonies increasing amount of adipocytes was revealed. The analysis of electron microscope microphotographs showed that in osteogenic cells differentiated under microgravity conditions, there is a reduction of the specific volume of a granular endoplasmic reticulum, Golgi's complex and quantity of nuclei reduction that indicates depression of the specific biosyntheses process intensity in cells. The increase of lysosomes and myelinic structures quantity is linked to organelles partial reduction. Consolidation of mitochondrias is an evidence of the cells’ energy metabolism disorder. In differentiated cells, disorganization and a cytoskeleton destruction was observed. Results showed that under microgravity conditions proliferative and differentiation (including osteogenic) potentialities of low-differentiated marrow stromal cells decreased, induction of their adipocytic differentiation was observes as well. Obtained results make a new contribution into gravitation sensitivity mechanisms understanding for stromal cells of the bone marrow which contain osteogenic cells- predecessors, features of the osteoporosis development.

  10. Determining the maximum charging currents of lithium-ion cells for small charge quantities

    NASA Astrophysics Data System (ADS)

    Grimsmann, F.; Gerbert, T.; Brauchle, F.; Gruhle, A.; Parisi, J.; Knipper, M.

    2017-10-01

    In order to optimize the operating parameters of battery management systems for electric and hybrid vehicles, great interest has been shown in achieving the maximum permissible charging currents during recuperation, without causing a cell damage due to lithium plating, in relation to the temperature, charge quantity and state of charge. One method for determining these recuperation currents is measuring the cell thickness, where excessively high charging currents can be detected by an irreversible increase in thickness. It is not possible to measure particularly small charge quantities by employing mechanic dial indicators, which have a limited resolution of 1 μm. This is why we developed a measuring setup that has a resolution limit of less than 10 nm using a high-resolution contactless inductance sensor. Our results show that the permissible charging current I can be approximated in relation to the charge quantity x by a correlating function I =a /√{(x) } which is compliant with the Arrhenius law. Small charge quantities therefore have an optimization potential for energy recovery during recuperation.

  11. Immune function and hematology of male cotton rats (Sigmodon hispidus) in response to food supplementation and methionine

    USGS Publications Warehouse

    Webb, R.E.; Leslie, David M.; Lochmiller, R.L.; Masters, R.E.

    2003-01-01

    We examined effects of supplementation of food quantity and quality (=enhanced methionine) on hematologic and immunologic parameters of wild, but enclosed, adult male cotton rats (Sigmodon hispidus) in north-central Oklahoma. Sheet metal enclosures were stocked with a high density of wild-caught cotton rats (160 animals/ha) and randomly assigned a treatment of no supplementation, mixed-ration supplementation or methionine-enhanced supplementation. Aside from small increases in counts of red blood cells and hematocrit levels, most indices of erythrocytic characteristics were not affected by supplementation with the mixed-ration or enhanced methionine. In contrast, platelet counts were highest in mixed-ration and methionine treatments and counts of total white blood cells were highest with methionine supplementation, albeit relative proportions of different leukocytes did not differ among treatments. Immunologically, neither delayed-type hypersensitivity response nor hemolytic-complement activity differed among treatments. Supplementation of food quantity and quality did not broadly affect hematologic parameters and immune function of male cotton rats, but enhanced platelet and leukocyte counts may confer advantages to overall health. Clarification of the role of such effects on population limitation or regulation requires additional research.

  12. 48 CFR 52.228-15 - Performance and Payment Bonds-Construction.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...) Definitions. As used in this clause— Original contract price means the award price of the contract; or, for requirements contracts, the price payable for the estimated total quantity; or, for indefinite-quantity contracts, the price payable for the specified minimum quantity. Original contract price does not include...

  13. 17 CFR Appendix A to Part 20 - Guidelines on Futures Equivalency

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... January 1 Total Notional Quantity = 6 months * 100,000 bbls/month = 600,000 bbls 1,000 bbl = 1 futures... Quantity = 6 contract months * 1,000,000 bushels/month = 6,000,000 bushels 5,000 bushels = 1 futures contract Therefore 6,000,000 bushels/5,000 bushels/contract = 1,200 futures equivalent contracts Total days...

  14. 17 CFR Appendix A to Part 20 - Guidelines on Futures Equivalency

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... January 1 Total Notional Quantity = 6 months * 100,000 bbls/month = 600,000 bbls 1,000 bbl = 1 futures... Quantity = 6 contract months * 1,000,000 bushels/month = 6,000,000 bushels 5,000 bushels = 1 futures contract Therefore 6,000,000 bushels/5,000 bushels/contract = 1,200 futures equivalent contracts Total days...

  15. 17 CFR Appendix A to Part 20 - Guidelines on Futures Equivalency

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... January 1 Total Notional Quantity = 6 months * 100,000 bbls/month = 600,000 bbls 1,000 bbl = 1 futures... Quantity = 6 contract months * 1,000,000 bushels/month = 6,000,000 bushels 5,000 bushels = 1 futures contract Therefore 6,000,000 bushels/5,000 bushels/contract = 1,200 futures equivalent contracts Total days...

  16. Identification of cytolytic vaginosis versus vulvovaginal candidiasis.

    PubMed

    Hu, Zhengqiang; Zhou, Wei; Mu, Liyuan; Kuang, Linghan; Su, Min; Jiang, Yongmei

    2015-04-01

    This study aimed to observe the morphological characteristic of vaginal discharge in patients with cytolytic vaginosis (CV) under the microscope and to identify it in patients with CV and in patients with vulvovaginal candidiasis (VVC). A total of 108 subjects including 21 healthy women, 33 patients with CV, and 54 patients with VVC were enrolled in the present morphological study. Vaginal discharge was collected and made into smear. The morphological characteristics of these vaginal smears with Gram staining were observed under the microscope. The smears were assessed for the quantity of lactobacilli, epithelial cell morphology, and the absence or presence of Candida species, Trichomonas vaginalis, and clue sells. First, the age, the level of education, and especially the status of pregnancy of patients with CV were significantly different from those of the patients with VVC. Second, the morphological characteristics of patients with CV consisted of overgrowth of lactobacilli, the presence of naked nuclei and fragments of the epithelial cells, a paucity of leukocytes, and the absence of Candida species and other pathogens. However, the morphological characteristic of patients with VVC consisted of the presence or absence of lactobacilli and the presence of normal epithelial cells, candidal spores, blastospores, hyphae, or other pathogens such as T. vaginalis and Gardnerella vaginalis. Both CV and VVC can be identified based on the quantity of lactobacilli, the morphology of the epithelial cells, and the absence or presence of Candida species and other pathogens, and the misdiagnosis of CV as VVC can be avoided.

  17. Modeling and Numerical Challenges in Eulerian-Lagrangian Computations of Shock-driven Multiphase Flows

    NASA Astrophysics Data System (ADS)

    Diggs, Angela; Balachandar, Sivaramakrishnan

    2015-06-01

    The present work addresses the numerical methods required for particle-gas and particle-particle interactions in Eulerian-Lagrangian simulations of multiphase flow. Local volume fraction as seen by each particle is the quantity of foremost importance in modeling and evaluating such interactions. We consider a general multiphase flow with a distribution of particles inside a fluid flow discretized on an Eulerian grid. Particle volume fraction is needed both as a Lagrangian quantity associated with each particle and also as an Eulerian quantity associated with the flow. In Eulerian Projection (EP) methods, the volume fraction is first obtained within each cell as an Eulerian quantity and then interpolated to each particle. In Lagrangian Projection (LP) methods, the particle volume fraction is obtained at each particle and then projected onto the Eulerian grid. Traditionally, EP methods are used in multiphase flow, but sub-grid resolution can be obtained through use of LP methods. By evaluating the total error and its components we compare the performance of EP and LP methods. The standard von Neumann error analysis technique has been adapted for rigorous evaluation of rate of convergence. The methods presented can be extended to obtain accurate field representations of other Lagrangian quantities. Most importantly, we will show that such careful attention to numerical methodologies is needed in order to capture complex shock interaction with a bed of particles. Supported by U.S. Department of Defense SMART Program and the U.S. Department of Energy PSAAP-II program under Contract No. DE-NA0002378.

  18. Testing of concrete by laser ablation

    DOEpatents

    Flesher, D.J.; Becker, D.L.; Beem, W.L.; Berry, T.C.; Cannon, N.S.

    1997-01-07

    A method is disclosed for testing concrete in a structure in situ, by: directing a succession of pulses of laser radiation at a point on the structure so that each pulse effects removal of a quantity of concrete and transfers energy to the concrete; detecting a characteristic of energy which has been transferred to the concrete; determining, separately from the detecting step, the total quantity of concrete removed by the succession of pulses; and calculating a property of the concrete on the basis of the detected energy characteristic and the determined total quantity of concrete removed. 1 fig.

  19. Statistical Characterization of Commercial 18650-Format Lithium-Ion Cell Thermal Runaway Behavior Based on Calorimetric Testing Results

    NASA Technical Reports Server (NTRS)

    Walker, William; Darst, John; Finegan, Donal; Bayles, Gary; Johnson, Kenneth; Darcy, Eric; Rickman, Steven

    2018-01-01

    Effective thermal management systems, designed to handle the impacts of thermal runaway (TR) and to prevent cell-to-cell propagation, are key to safe operation of lithium-ion (Li-ion) battery assemblies. Critical factors for optimizing these systems include the total energy released during a single cell TR event and the fraction of the total energy that is released through the cell casing vs. through the ejecta material. A unique calorimeter was utilized to examine the TR behavior of a statistically significant number of 18650-format Li-ion cells with varying manufacturers, chemistries, and capacities. The calorimeter was designed to contain the TR energy in a format conducive to discerning the fractions of energy released through the cell casing vs. through the ejecta material. Other benefits of this calorimeter included the ability to rapidly test of large quantities of cells and the intentional minimization of secondary combustion effects. High energy (270 Wh kg-1) and moderate energy (200 Wh kg-1) 18650 cells were tested. Some of the cells had an imbedded short circuit (ISC) device installed to aid in the examination of TR mechanisms under more realistic conditions. Other variations included cells with bottom vent (BV) features and cells with thin casings (0.22 l(1/4)m). After combining the data gathered with the calorimeter, a statistical approach was used to examine the probability of certain TR behavior, and the associated energy distributions, as a function of capacity, venting features, cell casing thickness and temperature.?

  20. Recycled Cell Phones - A Treasure Trove of Valuable Metals

    USGS Publications Warehouse

    Sullivan, Daniel E.

    2006-01-01

    This U.S. Geological Survey (USGS) Fact Sheet examines the potential value of recycling the metals found in obsolete cell phones. Cell phones seem ubiquitous in the United States and commonplace throughout most of the world. There were approximately 1 billion cell phones in use worldwide in 2002. In the United States, the number of cell phone subscribers increased from 340,000 in 1985 to 180 million in 2004. Worldwide, cell phone sales have increased from slightly more than 100 million units per year in 1997 to an estimated 779 million units per year in 2005. Cell phone sales are projected to exceed 1 billion units per year in 2009, with an estimated 2.6 billion cell phones in use by the end of that year. The U.S. Environmental Protection Agency estimated that, by 2005, as many as 130 million cell phones would be retired annually in the United States. The nonprofit organization INFORM, Inc., anticipated that, by 2005, a total of 500 million obsolete cell phones would have accumulated in consumers' desk drawers, store rooms, or other storage, awaiting disposal. Typically, cell phones are used for only 1 1/2 years before being replaced. Less than 1 percent of the millions of cell phones retired and discarded annually are recycled. When large numbers of cell phones become obsolete, large quantities of valuable metals end up either in storage or in landfills. The amount of metals potentially recoverable would make a significant addition to total metals recovered from recycling in the United States and would supplement virgin metals derived from mining.

  1. The endothelial sample size analysis in corneal specular microscopy clinical examinations.

    PubMed

    Abib, Fernando C; Holzchuh, Ricardo; Schaefer, Artur; Schaefer, Tania; Godois, Ronialci

    2012-05-01

    To evaluate endothelial cell sample size and statistical error in corneal specular microscopy (CSM) examinations. One hundred twenty examinations were conducted with 4 types of corneal specular microscopes: 30 with each BioOptics, CSO, Konan, and Topcon corneal specular microscopes. All endothelial image data were analyzed by respective instrument software and also by the Cells Analyzer software with a method developed in our lab. A reliability degree (RD) of 95% and a relative error (RE) of 0.05 were used as cut-off values to analyze images of the counted endothelial cells called samples. The sample size mean was the number of cells evaluated on the images obtained with each device. Only examinations with RE < 0.05 were considered statistically correct and suitable for comparisons with future examinations. The Cells Analyzer software was used to calculate the RE and customized sample size for all examinations. Bio-Optics: sample size, 97 ± 22 cells; RE, 6.52 ± 0.86; only 10% of the examinations had sufficient endothelial cell quantity (RE < 0.05); customized sample size, 162 ± 34 cells. CSO: sample size, 110 ± 20 cells; RE, 5.98 ± 0.98; only 16.6% of the examinations had sufficient endothelial cell quantity (RE < 0.05); customized sample size, 157 ± 45 cells. Konan: sample size, 80 ± 27 cells; RE, 10.6 ± 3.67; none of the examinations had sufficient endothelial cell quantity (RE > 0.05); customized sample size, 336 ± 131 cells. Topcon: sample size, 87 ± 17 cells; RE, 10.1 ± 2.52; none of the examinations had sufficient endothelial cell quantity (RE > 0.05); customized sample size, 382 ± 159 cells. A very high number of CSM examinations had sample errors based on Cells Analyzer software. The endothelial sample size (examinations) needs to include more cells to be reliable and reproducible. The Cells Analyzer tutorial routine will be useful for CSM examination reliability and reproducibility.

  2. BASiCS: Bayesian Analysis of Single-Cell Sequencing Data.

    PubMed

    Vallejos, Catalina A; Marioni, John C; Richardson, Sylvia

    2015-06-01

    Single-cell mRNA sequencing can uncover novel cell-to-cell heterogeneity in gene expression levels in seemingly homogeneous populations of cells. However, these experiments are prone to high levels of unexplained technical noise, creating new challenges for identifying genes that show genuine heterogeneous expression within the population of cells under study. BASiCS (Bayesian Analysis of Single-Cell Sequencing data) is an integrated Bayesian hierarchical model where: (i) cell-specific normalisation constants are estimated as part of the model parameters, (ii) technical variability is quantified based on spike-in genes that are artificially introduced to each analysed cell's lysate and (iii) the total variability of the expression counts is decomposed into technical and biological components. BASiCS also provides an intuitive detection criterion for highly (or lowly) variable genes within the population of cells under study. This is formalised by means of tail posterior probabilities associated to high (or low) biological cell-to-cell variance contributions, quantities that can be easily interpreted by users. We demonstrate our method using gene expression measurements from mouse Embryonic Stem Cells. Cross-validation and meaningful enrichment of gene ontology categories within genes classified as highly (or lowly) variable supports the efficacy of our approach.

  3. Differential loss of natural killer cell activity in patients with acute myocardial infarction and stable angina pectoris.

    PubMed

    Yan, Wenwen; Zhou, Lin; Wen, Siwan; Duan, Qianglin; Huang, Feifei; Tang, Yu; Liu, Xiaohong; Chai, Yongyan; Wang, Lemin

    2015-01-01

    To evaluate the activity of natural killer cells through their inhibitory and activating receptors and quantity in peripheral blood mononuclear cells extracted from patients with acute myocardial infarction, stable angina pectoris and the controls. 100 patients with myocardial infarction, 100 with stable angina, and 20 healthy volunteers were recruited into the study. 20 randomly chosen people per group were examined for the whole human genome microarray analysis to detect the gene expressions of all 40 inhibitory and activating natural killer cell receptors. Flow cytometry analysis was applied to all 200 patients to measure the quantity of natural killer cells. In myocardial infarction group, the mRNA expressions of six inhibitory receptors KIR2DL2, KIR3DL3, CD94, NKG2A, KLRB1, KLRG1, and eight activating receptors KIR2DS3, KIR2DS5, NKp30, NTB-A, CRACC, CD2, CD7 and CD96 were significantly down-regulated (P<0.05) compared with both angina patients and the controls. There was no statistical difference in receptor expressions between angina patients and control group. The quantity of natural killer cells was significantly decreased in both infarction and angina patients compared with normal range (P<0.001). The significant mRNAs down-regulation of several receptors in myocardial infarction group and reduction in the quantity of natural killer cells in both myocardial infarction and angina patients showed a quantitative loss and dysfunction of natural killer cells in myocardial infarction patients.

  4. 48 CFR 3.501-2 - General.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... submitted only for the total multiyear quantity; or (2) Priced options for additional quantities that... safeguards are available to the contracting officer to preclude recovery of buying-in losses (e.g...

  5. 48 CFR 3.501-2 - General.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... submitted only for the total multiyear quantity; or (2) Priced options for additional quantities that... safeguards are available to the contracting officer to preclude recovery of buying-in losses (e.g...

  6. 7 CFR 929.14 - Marketable quantity.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE CRANBERRIES GROWN IN STATES OF.... Marketable quantity means for a crop year the number of pounds of cranberries necessary to meet the total...

  7. 7 CFR 929.14 - Marketable quantity.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... AND ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE CRANBERRIES GROWN IN STATES OF.... Marketable quantity means for a crop year the number of pounds of cranberries necessary to meet the total...

  8. 7 CFR 929.14 - Marketable quantity.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... AND ORDERS; FRUITS, VEGETABLES, NUTS), DEPARTMENT OF AGRICULTURE CRANBERRIES GROWN IN STATES OF.... Marketable quantity means for a crop year the number of pounds of cranberries necessary to meet the total...

  9. 7 CFR 929.14 - Marketable quantity.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE CRANBERRIES GROWN IN STATES OF.... Marketable quantity means for a crop year the number of pounds of cranberries necessary to meet the total...

  10. 7 CFR 929.14 - Marketable quantity.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE CRANBERRIES GROWN IN STATES OF.... Marketable quantity means for a crop year the number of pounds of cranberries necessary to meet the total...

  11. 49 CFR 173.4a - Excepted quantities.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... ice), and lithium batteries and cells. (c) Inner packaging limits. The maximum quantity of hazardous materials in each inner packaging is limited to: (1) For toxic material with a Division 6.1 primary or... excepted quantities must meet the following: (1) Each inner receptacle must be constructed of plastic, or...

  12. BASiCS: Bayesian Analysis of Single-Cell Sequencing Data

    PubMed Central

    Vallejos, Catalina A.; Marioni, John C.; Richardson, Sylvia

    2015-01-01

    Single-cell mRNA sequencing can uncover novel cell-to-cell heterogeneity in gene expression levels in seemingly homogeneous populations of cells. However, these experiments are prone to high levels of unexplained technical noise, creating new challenges for identifying genes that show genuine heterogeneous expression within the population of cells under study. BASiCS (Bayesian Analysis of Single-Cell Sequencing data) is an integrated Bayesian hierarchical model where: (i) cell-specific normalisation constants are estimated as part of the model parameters, (ii) technical variability is quantified based on spike-in genes that are artificially introduced to each analysed cell’s lysate and (iii) the total variability of the expression counts is decomposed into technical and biological components. BASiCS also provides an intuitive detection criterion for highly (or lowly) variable genes within the population of cells under study. This is formalised by means of tail posterior probabilities associated to high (or low) biological cell-to-cell variance contributions, quantities that can be easily interpreted by users. We demonstrate our method using gene expression measurements from mouse Embryonic Stem Cells. Cross-validation and meaningful enrichment of gene ontology categories within genes classified as highly (or lowly) variable supports the efficacy of our approach. PMID:26107944

  13. Monitoring electrolyte concentrations in redox flow battery systems

    DOEpatents

    Chang, On Kok; Sopchak, David Andrew; Pham, Ai Quoc; Kinoshita, Kimio

    2015-03-17

    Methods, systems and structures for monitoring, managing electrolyte concentrations in redox flow batteries are provided by introducing a first quantity of a liquid electrolyte into a first chamber of a test cell and introducing a second quantity of the liquid electrolyte into a second chamber of the test cell. The method further provides for measuring a voltage of the test cell, measuring an elapsed time from the test cell reaching a first voltage until the test cell reaches a second voltage; and determining a degree of imbalance of the liquid electrolyte based on the elapsed time.

  14. Binding of glucosyltransferase and glucan synthesis by Streptococcus mutans and other bacteria.

    PubMed

    Hamada, S; Tai, S; Slade, H D

    1978-07-01

    Lyophilized and heat-treated cells from the seven serotypes of Streptococcus mutans were examined for their ability to bind added insoluble-product glucosyl-transferase (GTase) and to synthesize cell-associated glucan from [(14)C]sucrose. Lyophilized cells of serotypes a and g did not synthesize any more additional glucan than did the controls after exposure to GTase. These cells, however, synthesized four- to eightfold-greater quantities of glucan than did the cells of the remaining serotypes. Lyophilized cells of serotypes b, c, d, e, and f synthesized two- to threefold-greater quantities of glucan after exposure to GTase than did the controls without added enzyme. Lyophilized cells of serotypes a and g synthesized 6- to 10-fold-greater quantities of glucan than did heat-treated cells of the same strain after binding of GTase. Lyophilized cells of the remaining serotypes synthesized only 1.6- to 3.3-fold-greater quantities of glucan than did the heat-treated cells. These results demonstrate that heat treatment to inactivate cell-associated GTase does not create additional GTase binding sites in S. mutans and that serotypes a and g are considerably more active in cell-associated glucan synthesis than cells of the other five serotypes. Ten species of gram-positive and gram-negative bacteria from five genera which do not produce in vitro plaque synthesized 10- to 100-fold-less glucan than did the S. mutans strains after exposure to GTase. Of these species, S. sanguis, Actinomyces viscosus, and A. naeslundii synthesized the largest quantities of glucan. Three mutant strains of S. mutans which possess a reduced ability for in vitro adherence but do agglutinate with glucan or dextran synthesized only one-third as much glucan after binding of GTase as the control. These results are discussed in relation to in vitro and in vivo plaque development and the agglutination of S. mutans. The results support earlier findings which indicate that the presence of bacterial species other than S. mutans in smooth-surface dental plaque is due in part to contact of the cells with glucan in the developing plaque and not to the binding of cell-free GTase and the in situ synthesis of glucan. The results obtained with these representative strains of the seven serotypes of S. mutans may not apply to the same extent to other strains within the serotypes.

  15. Automated Decisional Model for Optimum Economic Order Quantity Determination Using Price Regressive Rates

    NASA Astrophysics Data System (ADS)

    Roşu, M. M.; Tarbă, C. I.; Neagu, C.

    2016-11-01

    The current models for inventory management are complementary, but together they offer a large pallet of elements for solving complex problems of companies when wanting to establish the optimum economic order quantity for unfinished products, row of materials, goods etc. The main objective of this paper is to elaborate an automated decisional model for the calculus of the economic order quantity taking into account the price regressive rates for the total order quantity. This model has two main objectives: first, to determine the periodicity when to be done the order n or the quantity order q; second, to determine the levels of stock: lighting control, security stock etc. In this way we can provide the answer to two fundamental questions: How much must be ordered? When to Order? In the current practice, the business relationships with its suppliers are based on regressive rates for price. This means that suppliers may grant discounts, from a certain level of quantities ordered. Thus, the unit price of the products is a variable which depends on the order size. So, the most important element for choosing the optimum for the economic order quantity is the total cost for ordering and this cost depends on the following elements: the medium price per units, the stock cost, the ordering cost etc.

  16. [Ecological distribution and antimicrobial effects of soil actinomycetes in artificial vegetation systems in Shazhuyu of Qinghai, China].

    PubMed

    Yang, Bin; Xue, Quan-hong; Chen, Zhan-quan; Guo, Zhi-ying; Zhang, Xiao-lu; Zhou, Yong-qiang; Xu, Ying-jun; Sun, De-fu

    2008-08-01

    In order to probe into the effects of artificial vegetation rehabilitation on soil actinomycetes, dilution plate and agar block methods were used to investigate the ecological distribution and antimicrobial effects of actinomycetes in sandy soil in Shazhuyu area of Qinghai after artificial vegetation restoration. The results showed that with the vegetation rehabilitation and the improvement of vegetation coverage on alpine sandy dry land, the quantity of soil actinomycetes increased significantly, being 145.4% higher in the grassland transferred from farmland than in sandy land. The quantity of soil Micromonospora in grassland transferred from farmland was about six times as much as that in sandy land. The average selection rate of antimicrobial actinomycetes was increased greatly, with the antimicrobial actinomycetes in the soil of grassland transferred from farmland, the antibacterial actinomycetes in the soil of natural grassland, and the pathogenic fungus resistant aetinomycetes in the soil of forestland being approximately 2, 3.2 and 1.5 times as much as those in the soil of sandy land, respectively. Vegetation coverage and soil nutrients had great influences on the quantities of actinomycetes and antimicrobial actinomycetes. The contents of soil organic matter and alkali-hydrolyzable nitrogen and the yield of fresh grasses had significant correlations with the quantities of actinomycetes (P < 0.01), and the content of soil organic matter and the yield of fresh grasses significantly correlated with the strain numbers of antimicrobial actinomycetes (P < 0.01). Furthermore, vegetation coverage and the contents of soil total nitrogen, total phosphorous, total potassium, total salt, and available potassium had significant correlations with the total quantities of actinomycetes, Streptomycetes, and Micromonospora (P < 0.05).

  17. Phytochemical and biological evaluation of some Sargassum species from Persian Gulf

    PubMed Central

    Mehdinezhad, Negin; Ghannadi, Alireza; Yegdaneh, Afsaneh

    2016-01-01

    Sea algae are widely consumed in the world. There are several seaweeds including brown algae which are authorized for human consumption. These plants contain important phytochemical constituents and have various potential biological activities. The present study investigated the presence of phytochemical constituents and total phenolic quantity of the seaweeds Sargassum angustifolium, Sargassum oligocystum and Sargassum boveanum. Cytotoxicity of seaweeds was tested against HT-29, HeLa and MCF-7 cell lines. Antioxidant potential of these 3 Sargassum species was also analyzed. Cytotoxicity was characterized by IC50 of human cancer cell lines using sulforhodamine assay. Antioxidant activities were evaluated using 2,2-diphenyl-1- picrylhydrazil. The analysis revealed that tannins, saponins, sterols and triterpenes were the most abundant compounds in these Sargassum species while cyanogenic and cardiac glycosides were the least ones. Sargassum angustifolium had the highest content of total phenolics (0.061 mg/g) and showed the highest antioxidant activity (IC50 = 0.231). Cytotoxic results showed that all species could inhibit cell growth effectively, especially MCF-7 cell line (IC50 = 67.3, 56.9, 60.4 for S. oligocystum, S. angustifolium and S. boveanum respectively). Considerable phytochemicals and moderate cytotoxic activity of S. angustifolium, S. oligocystum and S. boveanum make them appropriate candidate for further studies and identification of their bioactive principles. PMID:27499794

  18. Phytochemical and biological evaluation of some Sargassum species from Persian Gulf.

    PubMed

    Mehdinezhad, Negin; Ghannadi, Alireza; Yegdaneh, Afsaneh

    2016-01-01

    Sea algae are widely consumed in the world. There are several seaweeds including brown algae which are authorized for human consumption. These plants contain important phytochemical constituents and have various potential biological activities. The present study investigated the presence of phytochemical constituents and total phenolic quantity of the seaweeds Sargassum angustifolium, Sargassum oligocystum and Sargassum boveanum. Cytotoxicity of seaweeds was tested against HT-29, HeLa and MCF-7 cell lines. Antioxidant potential of these 3 Sargassum species was also analyzed. Cytotoxicity was characterized by IC50 of human cancer cell lines using sulforhodamine assay. Antioxidant activities were evaluated using 2,2-diphenyl-1- picrylhydrazil. The analysis revealed that tannins, saponins, sterols and triterpenes were the most abundant compounds in these Sargassum species while cyanogenic and cardiac glycosides were the least ones. Sargassum angustifolium had the highest content of total phenolics (0.061 mg/g) and showed the highest antioxidant activity (IC50 = 0.231). Cytotoxic results showed that all species could inhibit cell growth effectively, especially MCF-7 cell line (IC50 = 67.3, 56.9, 60.4 for S. oligocystum, S. angustifolium and S. boveanum respectively). Considerable phytochemicals and moderate cytotoxic activity of S. angustifolium, S. oligocystum and S. boveanum make them appropriate candidate for further studies and identification of their bioactive principles.

  19. Decreased IgA+ B cells population and IgA, IgG, IgM contents of the cecal tonsil induced by dietary high fluorine in broilers.

    PubMed

    Liu, Juan; Cui, Hengmin; Peng, Xi; Fang, Jing; Zuo, Zhicai; Deng, Junliang; Wang, Hesong; Wu, Bangyuan; Deng, Yuanxin; Wang, Kangping

    2013-05-02

    Fluoride is an environmental and industrial pollutant that affects various organs in humans and animals. The cecal tonsil is an important component of the mucosal immune system and performs important and unique immune functions. In the present study, we investigated the effects of dietary high fluorine on the quantities of IgA+ B cells in the cecal tonsil by immunohistochemistry, and the immunoglobulin A (IgA), immunoglobulin G (IgG) and immunoglobulin M (IgM) contents in the cecal tonsil by ELISA. A total of 280 one-day-old avian broilers were divided into four groups and fed on a corn-soybean basal diet as control diet (fluorine 22.6 mg/kg) or the same diet supplemented with 400, 800 and 1,200 mg/kg fluorine (high fluorine groups I, II and III) in the form of sodium fluoride, respectively, throughout a 42-day experimental period. The results showed that the quantities of IgA+ B cells were lower (p < 0.05 or p < 0.01) and the IgA, IgG, and IgM contents were decreased (p < 0.05 or p < 0.01) in high fluorine groups II and III in comparison with those of control group. It was concluded that dietary fluorine, in the 800-1,200 mg/kg range, could reduce the numbers of the IgA+ B cells and immunoglobulin contents in the cecal tonsil, implying the local mucosal immune function was ultimately impacted in broilers.

  20. Decreased IgA+ B Cells Population and IgA, IgG, IgM Contents of the Cecal Tonsil Induced by Dietary High Fluorine in Broilers

    PubMed Central

    Liu, Juan; Cui, Hengmin; Peng, Xi; Fang, Jing; Zuo, Zhicai; Deng, Junliang; Wang, Hesong; Wu, Bangyuan; Deng, Yuanxin; Wang, Kangping

    2013-01-01

    Fluoride is an environmental and industrial pollutant that affects various organs in humans and animals. The cecal tonsil is an important component of the mucosal immune system and performs important and unique immune functions. In the present study, we investigated the effects of dietary high fluorine on the quantities of IgA+ B cells in the cecal tonsil by immunohistochemistry, and the immunoglobulin A (IgA), immunoglobulin G (IgG) and immunoglobulin M (IgM) contents in the cecal tonsil by ELISA. A total of 280 one-day-old avian broilers were divided into four groups and fed on a corn-soybean basal diet as control diet (fluorine 22.6 mg/kg) or the same diet supplemented with 400, 800 and 1,200 mg/kg fluorine (high fluorine groups I, II and III) in the form of sodium fluoride, respectively, throughout a 42-day experimental period. The results showed that the quantities of IgA+ B cells were lower (p < 0.05 or p < 0.01) and the IgA, IgG, and IgM contents were decreased (p < 0.05 or p < 0.01) in high fluorine groups II and III in comparison with those of control group. It was concluded that dietary fluorine, in the 800–1,200 mg/kg range, could reduce the numbers of the IgA+ B cells and immunoglobulin contents in the cecal tonsil, implying the local mucosal immune function was ultimately impacted in broilers. PMID:23644827

  1. An Improved Total RNA Extraction Method for White Jelly Mushroom Tremella fuciformis Rich in Polysaccharides

    PubMed Central

    Zhu, Hanyu; Sun, Xueyan; Liu, Dongmei; Zheng, Liesheng; Chen, Liguo

    2017-01-01

    An improved method for extracting high quality and quantity RNA from a jelly mushroom and a dimorphic fungus—Tremella fuciformis which is especially rich in polysaccharides, is described. RNA was extracted from T. fuciformis mycelium M1332 and its parental monokaryotic yeast-like cells Y13 and Y32. The A260/280 and A260/230 ratios were both approximately 2, and the RNA integrity number was larger than 8.9. The yields of RNA were between 108 and 213 µg/g fresh wt. Downstream molecular applications including reverse transcriptional PCR and quantitative real-time PCR were also performed. This protocol is reliable and may be widely applicable for total RNA extraction from other jelly mushrooms or filamentous fungi rich in polysaccharides. PMID:29371814

  2. Performance of the Defense Acquisition System, 2015 Annual report

    DTIC Science & Technology

    2015-09-16

    Planned Total (From Start to Completion) Quantity-Adjusted Unit -Procurement Recurring-Flyaway Funding (1999–2014) NOTE: This measures biennial...PAUC is a different funding measure define by statute (10 United States Code [U.S.C.], sections 2430a and 2432) consisting of the total acquisition... measures that (unlike PAUC and APUC) are fully adjusted for any changes in procurement quantity. These results help compare procurement unit costs

  3. [Green space vegetation quantity in workshop area of Wuhan Iron and Steel Company].

    PubMed

    Chen, Fang; Zhou, Zhixiang; Wang, Pengcheng; Li, Haifang; Zhong, Yingfei

    2006-04-01

    Aimed at the complex community structure and higher fragmentation of urban green space, and based on the investigation of synusia structure and its coverage, this paper studied the vegetation quantity of ornamental green space in the workshop area of Wuhan Iron and Steel Company, with the help of GIS. The results showed that different life forms of ornamental plants in this area had a greater difference in their single leaf area and leaf area index (LAI), and the LAI was not only depended on single leaf area, but also governed by the shape of tree crown and the intensive degree of branches and leaves. The total vegetation quantity was 1 694.2 hm2, with the average LAI being 7.75, and the vegetation quantity of arbor-shrub-herb and arbor-shrub communities accounted for 79.7% and 92.3% of the total, respectively, reflecting that the green space structure was dominated by arbor species and by arbor-shrub-herb and arbor-shrub community types. Single layer-structured lawn had a less percentage, while the vegetation quantity of herb synusia accounted for 22.9% of the total, suggesting an afforestation characteristic of "making use of every bit of space" in the workshop area. The vegetation quantity of urban ornamental green space depended on the area of green space, its synusia structure, and the LAI and coverage of ornamental plants. In enlarging urban green space, ornamental plant species with high LAI should be selected, and community structure should be improved to have a higher vegetation quantity in urban area. To quantify the vegetation quantity of urban ornamental green space more accurately, synusia should be taken as the unit to measure the LAI of typical species, and the synusia structure and its coverage of different community types should be investigated with the help of remote sensing images and GIS.

  4. Development of recombinant adeno-associated virus vectors carrying small interfering RNA (shHec1)-mediated depletion of kinetochore Hec1 protein in tumor cells.

    PubMed

    Li, L; Yang, L; Scudiero, D A; Miller, S A; Yu, Z-X; Stukenberg, P T; Shoemaker, R H; Kotin, R M

    2007-05-01

    Transcript depletion using small interfering RNA (siRNA) technology represents a potentially valuable technique for the treatment of cancer. However, delivering therapeutic quantities of siRNA into solid tumors by chemical transfection is not feasible, whereas viral vectors efficiently transduce many human tumor cell lines. Yet producing sufficient quantities of viral vectors that elicit acute and selective cytotoxicity remains a major obstacle for preclinical and clinical trials. Using the invertebrate Spodoptera frugiperda (Sf9) cell line, we were able to produce high titer stocks of cytotoxic recombinant adeno-associated virus (rAAV) that express short hairpin RNA (shRNA) and that efficiently deplete Hec1 (highly expressed in cancer 1), or Kntc2 (kinetochore-associated protein 2), a kinetochore protein directly involved in kinetochore microtubule interactions, chromosome congression and spindle checkpoint signaling. Depletion of Hec1 protein results in persistent spindle checkpoint activation followed by cell death. Because Hec1 expression and activity are only present in mitotic cells, non-dividing cells were not affected by rAAV treatment. On the basis of the results of screening 56 human tumor cell lines with three different serotype vectors, we used a tumor xenograft model to test the effects in vivo. The effects of the shHec1 vector were evident in sectioned and stained tumors. The experiments with rAAV-shRNA vectors demonstrate the utility of producing vectors in invertebrate cells to obtain sufficient concentrations and quantities for solid tumor therapy. This addresses an important requirement for cancer gene therapy, to produce cytotoxic vectors in sufficient quantities and concentrations to enable quantitative transduction and selective killing of solid tumor cells.

  5. Floral nectar production and carbohydrate composition and the structure of receptacular nectaries in the invasive plant Bunias orientalis L. (Brassicaceae).

    PubMed

    Denisow, Bożena; Masierowska, Marzena; Antoń, Sebastian

    2016-11-01

    The data relating to the nectaries and nectar secretion in invasive Brassicacean taxa are scarce. In the present paper, the nectar production and nectar carbohydrate composition as well as the morphology, anatomy and ultrastructure of the floral nectaries in Bunias orientalis were investigated. Nectary glands were examined using light, fluorescence, scanning electron and transmission electron microscopy. The quantities of nectar produced by flowers and total sugar mass in nectar were relatively low. Total nectar carbohydrate production per 10 flowers averaged 0.3 mg. Nectar contained exclusively glucose (G) and fructose (F) with overall G/F ratio greater than 1. The flowers of B. orientalis have four nectaries placed at the base of the ovary. The nectarium is intermediate between two nectary types: the lateral and median nectary type (lateral and median glands stay separated) and the annular nectary type (both nectaries are united into one). Both pairs of glands represent photosynthetic type and consist of epidermis and glandular tissue. However, they differ in their shape, size, secretory activity, dimensions of epidermal and parenchyma cells, thickness of secretory parenchyma, phloem supply, presence of modified stomata and cuticle ornamentation. The cells of nectaries contain dense cytoplasm, plastids with starch grains and numerous mitochondria. Companion cells of phloem lack cell wall ingrowths. The ultrastructure of secretory cells indicates an eccrine mechanism of secretion. Nectar is exuded throughout modified stomata.

  6. Combining laser-assisted microdissection (LAM) and RNA-seq allows to perform a comprehensive transcriptomic analysis of epidermal cells of Arabidopsis embryo.

    PubMed

    Sakai, Kaori; Taconnat, Ludivine; Borrega, Nero; Yansouni, Jennifer; Brunaud, Véronique; Paysant-Le Roux, Christine; Delannoy, Etienne; Martin Magniette, Marie-Laure; Lepiniec, Loïc; Faure, Jean Denis; Balzergue, Sandrine; Dubreucq, Bertrand

    2018-01-01

    Genome-wide characterization of tissue- or cell-specific gene expression is a recurrent bottleneck in biology. We have developed a sensitive approach based on ultra-low RNA sequencing coupled to laser assisted microdissection for analyzing different tissues of the small Arabidopsis embryo. We first characterized the number of genes detected according to the quantity of tissue yield and total RNA extracted. Our results revealed that as low as 0.02 mm 2 of tissue and 50 pg of total RNA can be used without compromising the number of genes detected. The optimised protocol was used to compare the epidermal versus mesophyll cell transcriptomes of cotyledons at the torpedo-shaped stage of embryo development. The approach was validated by the recovery of well-known epidermal genes such AtML1 or AtPDF2 and genes involved in flavonoid and cuticular waxes pathways. Moreover, the interest and sensitivity of this approach were highlighted by the characterization of several transcription factors preferentially expressed in epidermal cells. This technical advance unlocks some current limitations of transcriptomic analyses and allows to investigate further and efficiently new biological questions for which only a very small amounts of cells need to be isolated. For instance, it paves the way to increasing the spatial accuracy of regulatory networks in developing small embryo of Arabidopsis or other plant tissues.

  7. Uracil-DNA Glycosylase in Base Excision Repair and Adaptive Immunity

    PubMed Central

    Doseth, Berit; Visnes, Torkild; Wallenius, Anders; Ericsson, Ida; Sarno, Antonio; Pettersen, Henrik Sahlin; Flatberg, Arnar; Catterall, Tara; Slupphaug, Geir; Krokan, Hans E.; Kavli, Bodil

    2011-01-01

    Genomic uracil is a DNA lesion but also an essential key intermediate in adaptive immunity. In B cells, activation-induced cytidine deaminase deaminates cytosine to uracil (U:G mispairs) in Ig genes to initiate antibody maturation. Uracil-DNA glycosylases (UDGs) such as uracil N-glycosylase (UNG), single strand-selective monofunctional uracil-DNA glycosylase 1 (SMUG1), and thymine-DNA glycosylase remove uracil from DNA. Gene-targeted mouse models are extensively used to investigate the role of these enzymes in DNA repair and Ig diversification. However, possible species differences in uracil processing in humans and mice are yet not established. To address this, we analyzed UDG activities and quantities in human and mouse cell lines and in splenic B cells from Ung+/+ and Ung−/− backcrossed mice. Interestingly, human cells displayed ∼15-fold higher total uracil excision capacity due to higher levels of UNG. In contrast, SMUG1 activity was ∼8-fold higher in mouse cells, constituting ∼50% of the total U:G excision activity compared with less than 1% in human cells. In activated B cells, both UNG and SMUG1 activities were at levels comparable with those measured for mouse cell lines. Moreover, SMUG1 activity per cell was not down-regulated after activation. We therefore suggest that SMUG1 may work as a weak backup activity for UNG2 during class switch recombination in Ung−/− mice. Our results reveal significant species differences in genomic uracil processing. These findings should be taken into account when mouse models are used in studies of uracil DNA repair and adaptive immunity. PMID:21454529

  8. Simulation-based optimization framework for reuse of agricultural drainage water in irrigation.

    PubMed

    Allam, A; Tawfik, A; Yoshimura, C; Fleifle, A

    2016-05-01

    A simulation-based optimization framework for agricultural drainage water (ADW) reuse has been developed through the integration of a water quality model (QUAL2Kw) and a genetic algorithm. This framework was applied to the Gharbia drain in the Nile Delta, Egypt, in summer and winter 2012. First, the water quantity and quality of the drain was simulated using the QUAL2Kw model. Second, uncertainty analysis and sensitivity analysis based on Monte Carlo simulation were performed to assess QUAL2Kw's performance and to identify the most critical variables for determination of water quality, respectively. Finally, a genetic algorithm was applied to maximize the total reuse quantity from seven reuse locations with the condition not to violate the standards for using mixed water in irrigation. The water quality simulations showed that organic matter concentrations are critical management variables in the Gharbia drain. The uncertainty analysis showed the reliability of QUAL2Kw to simulate water quality and quantity along the drain. Furthermore, the sensitivity analysis showed that the 5-day biochemical oxygen demand, chemical oxygen demand, total dissolved solids, total nitrogen and total phosphorous are highly sensitive to point source flow and quality. Additionally, the optimization results revealed that the reuse quantities of ADW can reach 36.3% and 40.4% of the available ADW in the drain during summer and winter, respectively. These quantities meet 30.8% and 29.1% of the drainage basin requirements for fresh irrigation water in the respective seasons. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Installation Restoration Program Records Search for Davis-Monthan Air Force Base, Arizona.

    DTIC Science & Technology

    1982-08-01

    inspection labs, and corrosion -2- control shops. These industrial operations generate varying quantities of waste oils , fuels , *solvents, and cleaners. The...standard procedures for the disposition of the majority of the waste oils , fuels , solvents, and cleaners has been (1) fire department training...and corrosion control shops. These industrial operations generate varying quantities of waste oils , fuels , solvents, and cleaners. The total quantity

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

    PubMed Central

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

    2013-01-01

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

  11. Children's multiplicative transformations of discrete and continuous quantities.

    PubMed

    Barth, Hilary; Baron, Andrew; Spelke, Elizabeth; Carey, Susan

    2009-08-01

    Recent studies have documented an evolutionarily primitive, early emerging cognitive system for the mental representation of numerical quantity (the analog magnitude system). Studies with nonhuman primates, human infants, and preschoolers have shown this system to support computations of numerical ordering, addition, and subtraction involving whole number concepts prior to arithmetic training. Here we report evidence that this system supports children's predictions about the outcomes of halving and perhaps also doubling transformations. A total of 138 kindergartners and first graders were asked to reason about the quantity resulting from the doubling or halving of an initial numerosity (of a set of dots) or an initial length (of a bar). Controls for dot size, total dot area, and dot density ensured that children were responding to the number of dots in the arrays. Prior to formal instruction in symbolic multiplication, division, or rational number, halving (and perhaps doubling) computations appear to be deployed over discrete and possibly continuous quantities. The ability to apply simple multiplicative transformations to analog magnitude representations of quantity may form a part of the toolkit that children use to construct later concepts of rational number.

  12. The Mass Flux of Non-renewable Energy for Humanity

    NASA Astrophysics Data System (ADS)

    Solomon, Edwin

    The global energy supply relies on non-renewable energy sources, coal, crude oil, and natural gas, along with nuclear power from uranium and these finite resources are located within the upper few kilometers of the Earth's crust. The total quantity of non-renewable energy resources consumed relative to the total quantity available is an essential question facing humanity. Analyses of energy consumption was conducted for the period 1800--2014 using data from the U. S. Energy Information Administration (EIA) and World Energy Production, 1800--1985 to determine the balance between non-renewable energy resources consumed and ultimately recoverable reserves. Annual energy consumption was plotted for each non-renewable resource followed by analyses to determine annual growth rates of consumption. Results indicated total energy consumption grew approximately exponentially 3.6% per year from 1800--1975 and was linear from 1975--2014. The ultimately recoverable reserves (URR) plus the total quantity consumed to date equals the total energy resource reserve prior to exploitation (7.15 x 1018 grams). Knowing the original resource quantity and the annual consumption and growth rates, we can forecast the duration of remaining resources using different scenarios. Alternatively, we can use population growth models and consumption trends to determine the per capita allocation trends and model that into the future. Alternative modeling of future resource allocation on a per capita bases suggests that resource lifetime may be significantly less than that predicted from consumption and production dynamics alone.

  13. Supply chain coordination with defective items and quantity discount

    NASA Astrophysics Data System (ADS)

    Lin, Hsien-Jen; Lin, Yu-Jen

    2014-12-01

    This study develops an integrated inventory system involving defective items and quantity discount for optimal pricing and ordering strategies. The model analysed in this study is one in which the buyer orders a quantity, the vendor produces more than buyer's order quantity in order to reduce set-up cost, and then he/she offers an all-units quantity discount to the buyer. Our objective is to determine the optimal order quantity, retail price, mark-up rate, and the number of shipments per production run from the vendor to the buyer, so that the entire supply chain joint total profit incurred has a maximum value. Furthermore, an algorithm of finding the optimal solution is developed. Numerical examples are provided to illustrate the theoretical results.

  14. 75 FR 81337 - Interim Final Regulation Deferring the Reporting Date for Certain Data Elements Required Under...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-27

    ... Meat processing facilities. 311411 Frozen fruit, juice, and vegetable manufacturing facilities. 311421... volume conversion factor. Y 98.256(m)(3) Only total quantity of crude oil plus the quantity of...

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

    PubMed

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

    2016-06-01

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

  16. Summer 2015 Internship Abstract

    NASA Technical Reports Server (NTRS)

    Smith, Courtney

    2015-01-01

    Green fluorescent protein (GFP) visually shows the expression of proteins by fluorescing when exposed to certain wavelengths of light. The GFP in this experiment was used to identify cells actively releasing viruses. The experiment focused on the effect of microgravity on the GFP expression of Akata B-cells infected with Epstein Barr Virus (EBV). Two flasks were prepared with 30 million cells each and two bioreactors were prepared with 50 million cells each. All four cultures were incubated for 16 days and fed every four days. Cellometer readings were taken on the feeding days to find cell size, viability, and GFP expression. In addition, the cells were treated with Propodium monoazide (PMA) and run through real time PCR to determine viral load on the feeding days. On the International Space Station air samples are taken to analyze the bacterial and fungal organisms in the air. The Sartorius Portable Airport is being investigated for potential use on the ISS to analyze for viral content in the air. Multiple samples were taken around Johnson Space Center building 37 and in Clear Lake Pediatric Clinic. The filter used was the gelatin membrane filter and the DNA was extracted directly from the filter. The DNA was then run through real time PCR for Varicella Zoster Virus (VZV) and EBV as well as GAPDH to test for the presence of DNA. The results so far have shown low DNA yield and no positive results for VZV or EBV. Further inquiry involves accurately replicating an atmosphere with high viral load from saliva as would be found on the ISS to run the air sampler in. Another line of research is stress hormones that may be correlated to the reactivation of latent viruses. The stress hormones from saliva samples are analyzed rather than blood samples. The quantity found in saliva shows the quantity of the hormones actually attached to cells and causing a reaction, whereas in the blood the quantity of hormones is the total amount released to cause a reaction. The particular hormones tested for were cortisol, alpha-amylase, and DHEA. The DHEA was very high in the two control samples tested. Regularly, samples came into the lab from local clinics to be tested for various viruses. Saliva, blood, body scrapes, and tears were received from the clinics and then run for VZV, EBV, and Human Simplex Virus 1 (HSV-1) with the results then reported back to the clinician. Blood, saliva, and urine from astronauts were also tested for viruses and logged. In addition, several cell cultures were brought up and grown, including adherent Human Lung Fibroblast (HFL) cells infected with VZV, and Akata B-cells infected with EBV.

  17. Exposure to tris(1,3-dichloro-2-propyl) phosphate for Two generations decreases fecundity of zebrafish at environmentally relevant concentrations.

    PubMed

    Zhang, Yongkang; Li, Meng; Li, Shuying; Wang, Qiangwei; Zhu, Guonian; Su, Guanyong; Letcher, Robert J; Liu, Chunsheng

    2018-05-14

    Previous studies reported that exposure to environmentally relevant concentrations of TDCIPP significantly decreased the number of cumulative eggs in zebrafish, but effects on the quantity of eggs and sperms remained unknown. Therefore, in this study, effects of TDCIPP on yolk diameter, surface morphology of eggs, sperm density and total motility were evaluated. First generation (F0) zebrafish larvae (Danio rerio) were exposed to 0, 50, 500 or 5000 ng/L tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) from 14 days post fertilization (dpf) to 120 dpf. The F0 generation of zebrafish were paired and F1 generation of embryos were collected and continuously exposed to the same concentrations of TDCIPP until 150 dpf. TDCIPP bioconcentration in the whole body as well as effects on survival and fecundity were evaluated in F1 generation. Exposure to TDCIPP resulted in an accumulation of the chemical and decreased survival of F1 generation of zebrafish. TDCIPP decreased cumulative production and changed surface morphology of eggs in females. In males, TDCIPP decreased total motility of sperm but did not affect sperm density. These effects on quality of egg and sperm might be responsible for the decreased hatching rates observed in cross mating experiments. Furthermore, TDCIPP exposure resulted in down-regulated gene expression related to gonadal development and maturation of germ cells in females or/and males, and the down-regulation was correlated to decreased fecundity. Taken together, the results suggested that exposure to TDCIPP could decrease the quantity of eggs and sperms by down-regulating the expression of genes related to gonadal development and maturation of germ cells in zebrafish. Copyright © 2018 Elsevier B.V. All rights reserved.

  18. Evidence of increased toxic Alexandrium tamarense dinoflagellate blooms in the eastern Bering Sea in the summers of 2004 and 2005.

    PubMed

    Natsuike, Masafumi; Saito, Rui; Fujiwara, Amane; Matsuno, Kohei; Yamaguchi, Atsushi; Shiga, Naonobu; Hirawake, Toru; Kikuchi, Takashi; Nishino, Shigeto; Imai, Ichiro

    2017-01-01

    The eastern Bering Sea has a vast continental shelf, which contains various endangered marine mammals and large fishery resources. Recently, high numbers of toxic A. tamarense resting cysts were found in the bottom sediment surface of the eastern Bering Sea shelf, suggesting that the blooms have recently occurred. However, little is known about the presence of A. tamarense vegetative cells in the eastern Bering Sea. This study's goals were to detect the occurrence of A. tamarense vegetative cells on the eastern Bering Sea shelf and to find a relationship between environmental factors and their presence. Inter-annual field surveys were conducted to detect A. tamarense cells and environmental factors, such as nutrients, salinity, chlorophyll a, and water temperature, along a transect line on the eastern Bering Sea shelf during the summers of 2004, 2005, 2006, 2009, 2012, and 2013. A. tamarense vegetative cells were detected during every sampling year, and their quantities varied greatly from year to year. The maximum cell densities of A. tamarense observed during the summers of 2004 and 2005 were much higher than the Paralytic shellfish poisoning warning levels, which are greater than 100-1,000 cells L-1, in other subarctic areas. Lower quantities of the species occurred during the summers of 2009, 2012, and 2013. A significant positive correlation between A. tamarense quantity and water temperature and significant negative correlations between A. tamarense quantity and nutrient concentrations (of phosphate, silicate, and nitrite and nitrate) were detected in every sampling period. The surface- and bottom-water temperatures varied significantly from year to year, suggesting that water temperatures, which have been known to affect the cell growth and cyst germination of A. tamarense, might have affected the cells' quantities in the eastern Bering Sea each summer. Thus, an increase in the Bering Sea shelf's water temperature during the summer will increase the frequency and scale of toxic blooms and the toxin contamination of plankton feeders. This poses serious threats to humans and the marine ecosystem.

  19. Standardization and assessment of cell culture media quantities in roller poly ethylene terephthalate bottles employed in the industrial rabies viral vaccine production.

    PubMed

    Jagannathan, S; Chaansha, S; Rajesh, K; Santhiya, T; Charles, C; Venkataramana, K N

    2009-09-15

    Vero cells are utilized for production of rabies vaccine. This study deals with the optimize quantity media require for the rabies vaccine production in the smooth roller surface. The rabies virus (Pasteur vaccine strain) is infected to monolayer of the various experimented bottles. To analyze the optimal quantity of media for the production of rabies viral harvest during the process of Vero cell derived rabies vaccine. The trials are started from 200 to 400 mL (PTARV-1, PTARV-2, PTARV-3, PTARV-4 and PTARV-5). The samples are taken in an appropriate time intervals for analysis of In Process Quality Control (IPQC) tests. The collected viral harvests are further processed to rabies vaccine in a pilot level and in addition to scale up an industrial level. Based on the evaluation the PTARV-2 (250 mL) show highly encouraging results for the Vero cell derived rabies vaccine production.

  20. Protocol to Isolate a Large Amount of Functional Oligodendrocyte Precursor Cells from the Cerebral Cortex of Adult Mice and Humans

    PubMed Central

    Medina-Rodríguez, Eva María; Arenzana, Francisco Javier; Bribián, Ana; de Castro, Fernando

    2013-01-01

    During development, oligodendrocytes are generated from oligodendrocyte precursor cells (OPCs), a cell type that is a significant proportion of the total cells (3-8%) in the adult central nervous system (CNS) of both rodents and humans. Adult OPCs are responsible for the spontaneous remyelination that occurs in demyelinating diseases like Multiple Sclerosis (MS) and they constitute an interesting source of cells for regenerative therapy in such conditions. However, there is little data regarding the neurobiology of adult OPCs isolated from mice since an efficient method to isolate them has yet to be established. We have designed a protocol to obtain viable adult OPCs from the cerebral cortex of different mouse strains and we have compared its efficiency with other well-known methods. In addition, we show that this protocol is also useful to isolate functional OPCs from human brain biopsies. Using this method we can isolate primary cortical OPCs in sufficient quantities so as to be able to study their survival, maturation and function, and to facilitate an evaluation of their utility in myelin repair. PMID:24303061

  1. Protocol to isolate a large amount of functional oligodendrocyte precursor cells from the cerebral cortex of adult mice and humans.

    PubMed

    Medina-Rodríguez, Eva María; Arenzana, Francisco Javier; Bribián, Ana; de Castro, Fernando

    2013-01-01

    During development, oligodendrocytes are generated from oligodendrocyte precursor cells (OPCs), a cell type that is a significant proportion of the total cells (3-8%) in the adult central nervous system (CNS) of both rodents and humans. Adult OPCs are responsible for the spontaneous remyelination that occurs in demyelinating diseases like Multiple Sclerosis (MS) and they constitute an interesting source of cells for regenerative therapy in such conditions. However, there is little data regarding the neurobiology of adult OPCs isolated from mice since an efficient method to isolate them has yet to be established. We have designed a protocol to obtain viable adult OPCs from the cerebral cortex of different mouse strains and we have compared its efficiency with other well-known methods. In addition, we show that this protocol is also useful to isolate functional OPCs from human brain biopsies. Using this method we can isolate primary cortical OPCs in sufficient quantities so as to be able to study their survival, maturation and function, and to facilitate an evaluation of their utility in myelin repair.

  2. Screening of antiproliferative effect of aqueous extracts of plant foods consumed in México on the breast cancer cell line MCF-7.

    PubMed

    García-Solís, Pablo; Yahia, Elhadi M; Morales-Tlalpan, Verónica; Díaz-Muñoz, Mauricio

    2009-01-01

    We evaluated the antiproliferative effect of aqueous extracts of 14 plant foods consumed in Mexico on the breast cancer cell line MCF-7. The plant foods used were avocado, black sapote, guava, mango, prickly pear cactus stems (called nopal in Mexico, cooked and raw), papaya, pineapple, four different cultivars of prickly pear fruit, grapes and tomato. β-Carotene, total phenolics and gallic acid contents and the antioxidant capacity, measured by the ferric reducing/antioxidant power and the 2,2-diphenyl-1,1-picrylhydrazyl radical scavenging assays, were analyzed in each aqueous extract. Only the papaya extract had a significant antiproliferative effect measured with the methylthiazolydiphenyl-tetrazolium bromide assay. We did not notice a relationship between the total phenolic content and the antioxidant capacity with antiproliferative effect. It is suggested that each extract of plant food has a unique combination of the quantity and quality of phytochemicals that could determine its biological activity. Besides, papaya represents a very interesting fruit to explore its antineoplastic activities.

  3. Summary of combined treatment under endoscope on 70 esophagus cancer inpatients

    NASA Astrophysics Data System (ADS)

    Wang, Cheng; Nong, Meilong; Li, Laisheng; Jia, Fang; Hao, Runchun

    1993-03-01

    We announce with satisfaction that combined treatment on 70 inpatients who suffered esophageal cancer in its middle or late course is perfectly successful. The combined methods include phototherapy, microwave therapy, and anticarcinogen local injection. The results are as follows: CR 3 cases, holds 4.3% of the total inpatients; PR 36 cases, 51.4%; MR 24 cases, 34.3%; NR 7 cases, 10%; the total effective rate 90%. Splendid results of treatment on enlarging the canal, improving dysphagia, and releasing obstruction have been obtained. The dysphagic grade increased from 66 to 148, the grade of esophagostenosis from 64 to 147, and the obstruction releasing rate is 69 out of 70 (that is 98.6%). The histological observation after treatment shows that 59/62 inpatients being reported as having cancer cells appear to have retrogression accompanied with a few or large quantities of necrotic cancer cells, and 3 inpatients were changed to negative reaction. No obvious poisoning or side effects arose. The combined treatment is more advantageous on those of old age or the physically weak and those who cannot stand for an operation, radiotherapy, or normal chemotherapy.

  4. Quantitative Differences in a Single Maternal Factor Determine Survival Probabilities among Drosophila Germ Cells.

    PubMed

    Slaidina, Maija; Lehmann, Ruth

    2017-01-23

    Germ cell death occurs in many species [1-3] and has been proposed as a mechanism by which the fittest, strongest, or least damaged germ cells are selected for transmission to the next generation. However, little is known about how the choice is made between germ cell survival and death. Here, we focus on the mechanisms that regulate germ cell survival during embryonic development in Drosophila. We find that the decision to die is a germ cell-intrinsic process linked to quantitative differences in germ plasm inheritance, such that higher germ plasm inheritance correlates with higher primordial germ cell (PGC) survival probability. We demonstrate that the maternal factor lipid phosphate phosphatase Wunen-2 (Wun2) regulates PGC survival in a dose-dependent manner. Since wun2 mRNA levels correlate with the levels of other maternal determinants at the single-cell level, we propose that Wun2 is used as a readout of the overall germ plasm quantity, such that only PGCs with the highest germ plasm quantity survive. Furthermore, we demonstrate that Wun2 and p53, another regulator of PGC survival, have opposite yet independent effects on PGC survival. Since p53 regulates cell death upon DNA damage and various cellular stresses, we hypothesize that together they ensure selection of the PGCs with highest germ plasm quantity and least cellular damage. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Are periprosthetic tissue reactions observed after revision of total disc replacement comparable to the reactions observed after total hip or knee revision surgery?

    PubMed Central

    Punt, Ilona M.; Austen, Shennah; Cleutjens, Jack P.M.; Kurtz, Steven M.; ten Broeke, René H.M.; van Rhijn, Lodewijk W.; Willems, Paul C.; van Ooij, André

    2011-01-01

    Study design Comparative study. Objective To compare periprosthetic tissue reactions observed after total disc replacement (TDR), total hip arthroplasty (THA) and total knee arthroplasty (TKA) revision surgery. Summary of background data Prosthetic wear debris leading to particle disease, followed by osteolysis, is often observed after THA and TKA. Although the presence of polyethylene (PE) particles and periprosthetic inflammation after TDR has been proven recently, osteolysis is rarely observed. The clinical relevance of PE wear debris in the spine remains poorly understood. Methods Number, size and shape of PE particles, as well as quantity and type of inflammatory cells in periprosthetic tissue retrieved during Charité TDR (n=22), THA (n=10) and TKA (n=4) revision surgery were compared. Tissue samples were stained with hematoxylin/eosin and examined by using light microscopy with bright field and polarized light. Results After THA, large numbers of PE particles <6 µm were observed, which were mainly phagocytosed by macrophages. The TKA group had a broad size range with many larger PE particles and more giant cells. In TDR, the size range was similar to that observed in TKA. However, the smallest particles were the most prevalent with 75% of the particles being <6 µm, as seen in revision THA. In TDR, both macrophages and giant cells were present with a higher number of macrophages. Conclusions Both small and large PE particles are present after TDR revision surgery compatible with both THA and TKA wear patterns. The similarities between periprosthetic tissue reactions in the different groups may give more insight in the clinical relevance of PE particles and inflammatory cells in the lumbar spine. The current findings may help to improve TDR design as applied from technologies previously developed in THA and TKA with the goal of a longer survival of TDR. PMID:21336235

  6. Developmental Changes is Expression of Beta-Adrenergic Receptors in Cultures of C2C12 Skeletal Muscle Cells

    NASA Technical Reports Server (NTRS)

    Young, Ronald B.; Bridge, K. Y.; Vaughn, J. R.

    2000-01-01

    beta-Adrenergic receptor (bAR) agonists have been reported to modulate growth in several mammalian and avian species, and bAR agonists presumably exert their physiological action on skeletal muscle cells through this receptor. Because of the importance of bAR regulation on muscle protein metabolism in muscle cells, the objectives of this study were to determine the developmental expression pattern of the bAR population in C2C12 skeletal muscle cells, and to analyze changes in both the quantity and isoform expression of the major muscle protein, myosin. The number of bAR in mononucleated C2C12 cells was approximately 8,000 bAR per cell, which is comparable with the population reported in several other nonmuscle cell types. However, the bar population increased after myoblast fusion to greater than 50,000 bAR per muscle cell equivalent. The reasons for this apparent over-expression of bAR in C2C12 cells is not known. The quantity of myosin also increased after C2C12 myoblast fusion, but the quantity of myosin was less than that reported in primary muscle cell cultures. Finally, at least five different isoforms of myosin heavy chain could be resolved in C2C12 cells, and three of these exhibited either increased or decreased developmental regulation relative to the others. Thus, C2C12 myoblasts undergo developmental regulation of bAR population and myosin heavy chain isoform expression.

  7. Automated cell disruption is a reliable and effective method of isolating RNA from fresh snap-frozen normal and malignant oral mucosa samples.

    PubMed

    Van der Vorst, Sébastien; Dekairelle, Anne-France; Irenge, Léonid; Hamoir, Marc; Robert, Annie; Gala, Jean-Luc

    2009-01-01

    This study compared automated vs. manual tissue grinding in terms of RNA yield obtained from oral mucosa biopsies. A total of 20 patients undergoing uvulectomy for sleep-related disorders and 10 patients undergoing biopsy for head and neck squamous cell carcinoma were enrolled in the study. Samples were collected, snap-frozen in liquid nitrogen, and divided into two parts of similar weight. Sample grinding was performed on one sample from each pair, either manually or using an automated cell disruptor. The performance and efficacy of each homogenization approach was compared in terms of total RNA yield (spectrophotometry, fluorometry), mRNA quantity [densitometry of specific TP53 amplicons and TP53 quantitative reverse-transcribed real-time PCR (qRT-PCR)], and mRNA quality (functional analysis of separated alleles in yeast). Although spectrophotometry and fluorometry results were comparable for both homogenization methods, TP53 expression values obtained by amplicon densitometry and qRT-PCR were significantly and consistently better after automated homogenization (p<0.005) for both uvula and tumor samples. Functional analysis of separated alleles in yeast results was better with the automated technique for tumor samples. Automated tissue homogenization appears to be a versatile, quick, and reliable method of cell disruption and is especially useful in the case of small malignant samples, which show unreliable results when processed by manual homogenization.

  8. Characteristics and in vitro Anti-diabetic Properties of the Korean Rice Wine, Makgeolli Fermented with Laminaria japonica.

    PubMed

    Choi, Jae-Suk; Seo, Hyo Ju; Lee, Yu-Ri; Kwon, Su-Jung; Moon, Sun Hwa; Park, Sun-Mee; Sohn, Jae Hak

    2014-06-01

    New in vitro anti-diabetes makgeolli was produced from rice by adding various quantities of Laminaria japonica, and the fermentation characteristics of the L. japonica makgeolli during the fermentation process were investigated. The contents of alcohol and reducing sugar, and viable count of yeast, of L. japonica makgeolli were not significantly changed when the proportion of L. japonica was increased. The total acid content decreased with an increase in L. japonica concentration; the pH and total bacterial cell count increased in proportion with the increase in L. japonica concentration. The L. japonica makgeolli contents of free sugars, such as fructose, glucose, and sucrose, and of organic acids, such as acetic acid, citric acid, succinic acid, and lactic acid, were altered during fermentation and showed various patterns. The effects of the quantity of L. japonica added on the acceptability and anti-diabetes activities of L. japonica makgeolli were also investigated. In a sensory evaluation, L. japonica makgeolli brewed by adding 2.5 or 5% L. japonica to the mash showed the best overall acceptability; the 12.5% L. japonica sample was least favored due to its seaweed flavor. L. japonica addition did not increase the peroxynitrite-scavenging activity of makgeolli. L. japonica makgeolli showed potent anti-diabetes activity, particularly that containing >7.5% L. japonica. Therefore, L. japonica makgeolli may represent a new functional makgeolli with anti-diabetes properties.

  9. Technology commercialization cost model and component case study

    NASA Astrophysics Data System (ADS)

    1991-12-01

    Fuel cells seem poised to emerge as a clean, efficient, and cost competitive source of fossil fuel based electric power and thermal energy. Sponsors of fuel cell technology development need to determine the validity and the attractiveness of a technology to the market in terms of meeting requirements and providing value which exceeds the total cost of ownership. Sponsors of fuel cell development have addressed this issue by requiring the developers to prepare projections of the future production cost of their fuel cells in commercial quantities. These projected costs, together with performance and life projections, provide a preliminary measure of the total value and cost of the product to the customer. Booz-Allen & Hamilton Inc. and Michael A. Cobb & Company have been retained in several assignments over the years to audit these cost projections. The audits have gone well beyond a simple review of the numbers. They have probed the underlying technical and financial assumptions, the sources of data on material and equipment costs, and explored issues such as the realistic manufacturing yields which can be expected in various processes. Based on the experience gained from these audits, DOE gave Booz-Allen and Michael A. Cobb & company the task to develop a criteria to be used in the execution of future fuel cell manufacturing cost studies. It was thought that such a criteria would make it easier to execute such studies in the future as well as to cause such studies to be more understandable and comparable.

  10. Evaluation of STAT5A Gene Expression in Aflatoxin B1 Treated Bovine Mammary Epithelial Cells

    PubMed Central

    Forouharmehr, Ali; Harkinezhad, Taher; Qasemi-Panahi, Babak

    2013-01-01

    Purpose: Aflatoxin B1 (AFB1) is a potent mycotoxin which has been produced by fungi such as Aspergillus flavus and Aspergillus parasiticus as secondary metabolites due to their growth on food stuffs and induces hepatocellular carcinoma in many animal species, including humans. In the present study, the effect of AFB1 on STAT5A gene expression was investigated in bovine mammary epithelial cells using real time RT-PCR. Methods: Bovine mammary epithelial cells were seeded in a 24-well culture plate for three-dimensional (3D) culture in Matrigel matrix. After 21 days of 3D culture and reaching the required number of cells, cells were treated with AFB1 and incubated for 8 h. For real time PCR reaction, total RNA from the cultured and treated cells was extracted and used for complementary DNA synthesis. Results: The expression of STAT5A gene was significantly down regulated by AFB1 in dose- dependent manner and led to the reduction of proliferation and differentiation of epithelial cells, which has direct effect in milk protein quantity and quality. Conclusion: According to the results, it seems that down regulation of STAT5A gene in AFB1-treated cells maybe due to DNA damage induced by AFB1 in bovine mammary epithelial cells. PMID:24312879

  11. 78 FR 50045 - 36(b)(1) Arms Sales Notification

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-16

    ... Total $54 million (iii) Description and Quantity or Quantities of Articles or Services under... Commissions, Fee, etc: None (vii) Sensitivity of Technology Contained in the Defense Article or Defense... guidance kits in its inventory. The proposed sale of this equipment will not alter the basic military...

  12. 40 CFR 61.342 - Standards: General.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standard for Benzene Waste... benzene quantity from facility waste is less than 10 megagrams per year (Mg/yr) (11 ton/yr) shall be exempt from the requirements of paragraphs (b) and (c) of this section. The total annual benzene quantity...

  13. 40 CFR 61.342 - Standards: General.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standard for Benzene Waste... benzene quantity from facility waste is less than 10 megagrams per year (Mg/yr) (11 ton/yr) shall be exempt from the requirements of paragraphs (b) and (c) of this section. The total annual benzene quantity...

  14. 40 CFR 61.342 - Standards: General.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...) NATIONAL EMISSION STANDARDS FOR HAZARDOUS AIR POLLUTANTS National Emission Standard for Benzene Waste... benzene quantity from facility waste is less than 10 megagrams per year (Mg/yr) (11 ton/yr) shall be exempt from the requirements of paragraphs (b) and (c) of this section. The total annual benzene quantity...

  15. 7 CFR 929.49 - Marketable quantity, allotment percentage, and annual allotment.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...) AGRICULTURAL MARKETING SERVICE (Marketing Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF... history, established pursuant to § 929.48. Such allotment percentage shall be established by the Secretary and shall equal the marketable quantity divided by the total of all growers' sales histories including...

  16. Use of bioreactors for culturing human retinal organoids improves photoreceptor yields.

    PubMed

    Ovando-Roche, Patrick; West, Emma L; Branch, Matthew J; Sampson, Robert D; Fernando, Milan; Munro, Peter; Georgiadis, Anastasios; Rizzi, Matteo; Kloc, Magdalena; Naeem, Arifa; Ribeiro, Joana; Smith, Alexander J; Gonzalez-Cordero, Anai; Ali, Robin R

    2018-06-13

    The use of human pluripotent stem cell-derived retinal cells for cell therapy strategies and disease modelling relies on the ability to obtain healthy and organised retinal tissue in sufficient quantities. Generating such tissue is a lengthy process, often taking over 6 months of cell culture, and current approaches do not always generate large quantities of the major retinal cell types required. We adapted our previously described differentiation protocol to investigate the use of stirred-tank bioreactors. We used immunohistochemistry, flow cytometry and electron microscopy to characterise retinal organoids grown in standard and bioreactor culture conditions. Our analysis revealed that the use of bioreactors results in improved laminar stratification as well as an increase in the yield of photoreceptor cells bearing cilia and nascent outer-segment-like structures. Bioreactors represent a promising platform for scaling up the manufacture of retinal cells for use in disease modelling, drug screening and cell transplantation studies.

  17. Update on International Space Station Nickel-Hydrogen Battery On-Orbit Performance

    NASA Technical Reports Server (NTRS)

    Dalton, Penni; Cohen, Fred

    2003-01-01

    International Space Station (ISS) Electric Power System (EPS) utilizes Nickel-Hydrogen (Ni-H2) batteries as part of its power system to store electrical energy. The batteries are charged during insolation and discharged during eclipse. The batteries are designed to operate at a 35% depth of discharge (DOD) maximum during normal operation. Thirty-eight individual pressure vessel (IPV) Ni-H2 battery cells are series-connected and packaged in an Orbital Replacement Unit (ORU). Two ORUs are series-connected utilizing a total of 76 cells, to form one battery. The ISS is the first application for low earth orbit (LEO) cycling of this quantity of series-connected cells. The P6 (Port) Integrated Equipment Assembly (IEA) containing the initial ISS high-power components was successfully launched on November 30, 2000. The IEA contains 12 Battery Subassembly ORUs (6 batteries) that provide station power during eclipse periods. This paper will discuss the battery performance data after two and a half years of cycling.

  18. International Space Station Nickel-Hydrogen Battery On-Orbit Performance

    NASA Technical Reports Server (NTRS)

    Dalton, Penni; Cohen, Fred

    2002-01-01

    International Space Station (ISS) Electric Power System (EPS) utilizes Nickel-Hydrogen (Ni-H2) batteries as part of its power system to store electrical energy. The batteries are charged during insolation and discharged during eclipse. The batteries are designed to operate at a 35 percent depth of discharge (DOD) maximum during normal operation. Thirty-eight individual pressure vessel (IPV) Ni-H2 battery cells are series-connected and packaged in an Orbital Replacement Unit (ORU). Two ORUs are series-connected utilizing a total of 76 cells to form one battery. The ISS is the first application for low earth orbit (LEO) cycling of this quantity of series-connected cells. The P6 (Port) Integrated Equipment Assembly (IEA) containing the initial ISS high-power components was successfully launched on November 30, 2000. The IEA contains 12 Battery Subassembly ORUs (6 batteries) that provide station power during eclipse periods. This paper will discuss the battery performance data after eighteen months of cycling.

  19. Modelling the balance between quiescence and cell death in normal and tumour cell populations.

    PubMed

    Spinelli, Lorenzo; Torricelli, Alessandro; Ubezio, Paolo; Basse, Britta

    2006-08-01

    When considering either human adult tissues (in vivo) or cell cultures (in vitro), cell number is regulated by the relationship between quiescent cells, proliferating cells, cell death and other controls of cell cycle duration. By formulating a mathematical description we see that even small alterations of this relationship may cause a non-growing population to start growing with doubling times characteristic of human tumours. Our model consists of two age structured partial differential equations for the proliferating and quiescent cell compartments. Model parameters are death rates from and transition rates between these compartments. The partial differential equations can be solved for the steady-age distributions, giving the distribution of the cells through the cell cycle, dependent on specific model parameter values. Appropriate formulas can then be derived for various population characteristic quantities such as labelling index, proliferation fraction, doubling time and potential doubling time of the cell population. Such characteristic quantities can be estimated experimentally, although with decreasing precision from in vitro, to in vivo experimental systems and to the clinic. The model can be used to investigate the effects of a single alteration of either quiescence or cell death control on the growth of the whole population and the non-trivial dependence of the doubling time and other observable quantities on particular underlying cell cycle scenarios of death and quiescence. The model indicates that tumour evolution in vivo is a sequence of steady-states, each characterised by particular death and quiescence rate functions. We suggest that a key passage of carcinogenesis is a loss of the communication between quiescence, death and cell cycle machineries, causing a defect in their precise, cell cycle dependent relationship.

  20. 46 CFR 193.15-5 - Quantity, pipe sizes, and discharge rates.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... VESSELS FIRE PROTECTION EQUIPMENT Carbon Dioxide and Clean Agent Extinguishing Systems, Details § 193.15-5 Quantity, pipe sizes, and discharge rates. (a) General. The amount of carbon dioxide required for each.... A separate supply of carbon dioxide need not be provided for each space protected. The total...

  1. 46 CFR 193.15-5 - Quantity, pipe sizes, and discharge rates.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... VESSELS FIRE PROTECTION EQUIPMENT Carbon Dioxide and Clean Agent Extinguishing Systems, Details § 193.15-5 Quantity, pipe sizes, and discharge rates. (a) General. The amount of carbon dioxide required for each.... A separate supply of carbon dioxide need not be provided for each space protected. The total...

  2. 77 FR 46699 - Honey From the People's Republic of China: Preliminary Results of Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-06

    ... quantity and value, its separate rate status, structure and affiliations, sales process, accounting and... quantity and value, separate rate status, structure and affiliations, sales process, accounting and... (CIT August 10, 2009) (''Commerce may, of course, begin its total AFA selection process by defaulting...

  3. 77 FR 65185 - 36(b)(1) Arms Sales Notification

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-25

    ... million Total $60 million (iii) Description and Quantity or Quantities of Articles or Services under... Technology Contained in the Defense Article or Defense Services Proposed to be Sold: See Annex attached... integrate the Helmet Mounted Cueing System. The software algorithms are the most sensitive portion of the...

  4. 78 FR 62600 - 36(b)(1) Arms Sales Notification

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-22

    ... million Total $68 million (iii) Description and Quantity or Quantities of Articles or Services Under... Technology Contained in the Defense Article or Defense Services Proposed To Be Sold: See Annex attached... integrate the Helmet Mounted Cueing System. The software algorithms are the most sensitive portion of the...

  5. Improvement of Cell Survival During Human Pluripotent Stem Cell Definitive Endoderm Differentiation

    PubMed Central

    Wang, Han; Luo, Xie; Yao, Li; Lehman, Donna M.

    2015-01-01

    Definitive endoderm (DE) is a vital precursor for internal organs such as liver and pancreas. Efficient protocol to differentiate human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs) to DE is essential for regenerative medicine and for modeling diseases; yet, poor cell survival during DE differentiation remains unsolved. In this study, our use of B27 supplement in modified differentiation protocols has led to a substantial improvement. We used an SOX17-enhanced green fluorescent protein (eGFP) reporter hESC line to compare and modify established DE differentiation protocols. Both total live cell numbers and the percentages of eGFP-positive cells were used to assess differentiation efficiency. Among tested protocols, three modified protocols with serum-free B27 supplement were developed to generate a high number of DE cells. Massive cell death was avoided during DE differentiation and the percentage of DE cells remained high. When the resulting DE cells were further differentiated toward the pancreatic lineage, the expression of pancreatic-specific markers was significantly increased. Similar high DE differentiation efficiency was observed in H1 hESCs and iPSCs through the modified protocols. In B27 components, bovine serum albumin was found to facilitate DE differentiation and cell survival. Using our modified DE differentiation protocols, satisfactory quantities of quality DE can be produced as primary material for further endoderm lineage differentiation. PMID:26132288

  6. Tumor inhibitory T cell immunity may be largely a transplantation artifact not necessarily dependent upon a lack of Tregs.

    PubMed

    Prehn, Richmond T; Prehn, Liisa M

    2013-06-25

    There exists a very large literature suggesting that T cells come in a variety of species and that without the action of Tregs tumors would seldom survive inhibition by T cell effectors. We believe that much of the evidence supporting the role of Tregs in cancer is compatible with a perhaps simpler hypothesis based upon the demonstration that that small quantities of effector T cells tend to stimulate tumors while larger quantities of seemingly the same cells are inhibitory (an hormesis-like effect). This possibility seems to destroy much of the need to postulate a role for T cell suppressors (Tregs) in cancer, but the exposure of effector T cells to antigen may convert them into Tregs (Tregs do exist). Furthermore, many other data suggest the possibility that immune inhibition of cancer could be a laboratory artifact seldom if ever seen in unmodified nature.

  7. Tumor inhibitory T cell immunity may be largely a transplantation artifact not necessarily dependent upon a lack of Tregs

    PubMed Central

    2013-01-01

    There exists a very large literature suggesting that T cells come in a variety of species and that without the action of Tregs tumors would seldom survive inhibition by T cell effectors. We believe that much of the evidence supporting the role of Tregs in cancer is compatible with a perhaps simpler hypothesis based upon the demonstration that that small quantities of effector T cells tend to stimulate tumors while larger quantities of seemingly the same cells are inhibitory (an hormesis-like effect). This possibility seems to destroy much of the need to postulate a role for T cell suppressors (Tregs) in cancer, but the exposure of effector T cells to antigen may convert them into Tregs (Tregs do exist). Furthermore, many other data suggest the possibility that immune inhibition of cancer could be a laboratory artifact seldom if ever seen in unmodified nature. PMID:23800315

  8. Highly sensitive three-dimensional interdigitated microelectrode for microparticle detection using electrical impedance spectroscopy

    NASA Astrophysics Data System (ADS)

    Chang, Fu-Yu; Chen, Ming-Kun; Wang, Min-Haw; Jang, Ling-Sheng

    2016-02-01

    Cell impedance analysis is widely used for monitoring biological and medical reactions. In this study, a highly sensitive three-dimensional (3D) interdigitated microelectrode (IME) with a high aspect ratio on a polyimide (PI) flexible substrate was fabricated for microparticle detection (e.g. cell quantity detection) using electroforming and lithography technology. 3D finite element simulations were performed to compare the performance of the 3D IME (in terms of sensitivity and signal-to-noise ratio) to that of a planar IME for particles in the sensing area. Various quantities of particles were captured in Dulbecco’s modified Eagle medium and their impedances were measured. With the 3D IME, the particles were arranged in the gap, not on the electrode, avoiding the noise due to particle position. For the maximum particle quantities, the results show that the 3D IME has at least 5-fold higher sensitivity than that of the planar IME. The trends of impedance magnitude and phase due to particle quantity were verified using the equivalent circuit model. The impedance (1269 Ω) of 69 particles was used to estimate the particle quantity (68 particles) with 98.6% accuracy using a parabolic regression curve at 500 kHz.

  9. Uncertainty analysis of thermal quantities measurement in a centrifugal compressor

    NASA Astrophysics Data System (ADS)

    Hurda, Lukáš; Matas, Richard

    2017-09-01

    Compressor performance characteristics evaluation process based on the measurement of pressure, temperature and other quantities is examined to find uncertainties for directly measured and derived quantities. CFD is used as a tool to quantify the influences of different sources of uncertainty of measurements for single- and multi-thermocouple total temperature probes. The heat conduction through the body of the thermocouple probe and the heat-up of the air in the intake piping are the main phenomena of interest.

  10. B-2 Extremely High Frequency SATCOM and Computer Increment 1 (B-2 EHF Inc 1)

    DTIC Science & Technology

    2015-12-01

    Confidence Level Confidence Level of cost estimate for current APB: 55% This APB reflects cost and funding data based on the B-2 EHF Increment I SCP...This cost estimate was quantified at the Mean (~55%) confidence level . Total Quantity Quantity SAR Baseline Production Estimate Current APB...Production Estimate Econ Qty Sch Eng Est Oth Spt Total 33.624 -0.350 1.381 0.375 0.000 -6.075 0.000 -0.620 -5.289 28.335 Current SAR Baseline to Current

  11. A Dynamic Bayesian Network Model for the Production and Inventory Control

    NASA Astrophysics Data System (ADS)

    Shin, Ji-Sun; Takazaki, Noriyuki; Lee, Tae-Hong; Kim, Jin-Il; Lee, Hee-Hyol

    In general, the production quantities and delivered goods are changed randomly and then the total stock is also changed randomly. This paper deals with the production and inventory control using the Dynamic Bayesian Network. Bayesian Network is a probabilistic model which represents the qualitative dependence between two or more random variables by the graph structure, and indicates the quantitative relations between individual variables by the conditional probability. The probabilistic distribution of the total stock is calculated through the propagation of the probability on the network. Moreover, an adjusting rule of the production quantities to maintain the probability of a lower limit and a ceiling of the total stock to certain values is shown.

  12. Evaluation of neutron thermalization parameters and benchmark reactor calculations using a synthetic scattering function for molecular gases

    NASA Astrophysics Data System (ADS)

    Gillette, V. H.; Patiño, N. E.; Granada, J. R.; Mayer, R. E.

    1989-08-01

    Using a synthetic incoherent scattering function which describes the interaction of neutrons with molecular gases we provide analytical expressions for zero- and first-order scattering kernels, σ0( E0 → E), σ1( E0 → E), and total cross section σ0( E0). Based on these quantities, we have performed calculations of thermalization parameters and transport coefficients for H 2O, D 2O, C 6H 6 and (CH 2) n at room temperature. Comparison of such values with available experimental data and other calculations is satisfactory. We also generated nuclear data libraries for H 2O with 47 thermal groups at 300 K and performed some benchmark calculations ( 235U, 239Pu, PWR cell and typical APWR cell); the resulting reactivities are compared with experimental data and ENDF/B-IV calculations.

  13. The production of multiprotein complexes in insect cells using the baculovirus expression system.

    PubMed

    Abdulrahman, Wassim; Radu, Laura; Garzoni, Frederic; Kolesnikova, Olga; Gupta, Kapil; Osz-Papai, Judit; Berger, Imre; Poterszman, Arnaud

    2015-01-01

    The production of a homogeneous protein sample in sufficient quantities is an essential prerequisite not only for structural investigations but represents also a rate-limiting step for many functional studies. In the cell, a large fraction of eukaryotic proteins exists as large multicomponent assemblies with many subunits, which act in concert to catalyze specific activities. Many of these complexes cannot be obtained from endogenous source material, so recombinant expression and reconstitution are then required to overcome this bottleneck. This chapter describes current strategies and protocols for the efficient production of multiprotein complexes in large quantities and of high quality, using the baculovirus/insect cell expression system.

  14. 49 CFR 173.4a - Excepted quantities.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... of withstanding without leakage the pressure differential specified in § 173.27(c) of this part. (b... ice), and lithium batteries and cells. (c) Inner packaging limits. The maximum quantity of hazardous..., rigid outer packaging. (5) Placement of the material in the package or packing different materials in...

  15. Human Milk MicroRNA and Total RNA Differ Depending on Milk Fractionation

    PubMed Central

    Alsaweed, Mohammed; Hepworth, Anna R.; Lefèvre, Christophe; Hartmann, Peter E.; Geddes, Donna T.

    2015-01-01

    ABSTRACT MicroRNA have been recently discovered in human milk signifying potentially important functions for both the lactating breast and the infant. Whilst human milk microRNA have started to be explored, little data exist on the evaluation of sample processing, and analysis to ensure that a full spectrum of microRNA can be obtained. Human milk comprises three main fractions: cells, skim milk, and lipids. Typically, the skim milk fraction has been measured in isolation despite evidence that the lipid fraction may contain more microRNA. This study aimed to standardize isolation of microRNA and total RNA from all three fractions of human milk to determine the most appropriate sampling and analysis procedure for future studies. Three different methods from eight commercially available kits were tested for their efficacy in extracting total RNA and microRNA from the lipid, skim, and cell fractions of human milk. Each fraction yielded different concentrations of RNA and microRNA, with the highest quantities found in the cell and lipid fractions, and the lowest in skim milk. The column‐based phenol‐free method was the most efficient extraction method for all three milk fractions. Two microRNAs were expressed and validated in the three milk fractions by qPCR using the three recommended extraction kits for each fraction. High expression levels were identified in the skim and lipid milk factions for these microRNAs. These results suggest that careful consideration of both the human milk sample preparation and extraction protocols should be made prior to embarking upon research in this area. J. Cell. Biochem. 116: 2397–2407, 2015. © 2015 The Authors. Journal of Cellular Biochemistry Published by Wiley Periodicals, Inc. PMID:25925799

  16. Viral Impacts on Total Abundance and Clonal Composition of the Harmful Bloom-Forming Phytoplankton Heterosigma akashiwo

    PubMed Central

    Tarutani, Kenji; Nagasaki, Keizo; Yamaguchi, Mineo

    2000-01-01

    Recent observations that viruses are very abundant and biologically active components in marine ecosystems suggest that they probably influence various biogeochemical and ecological processes. In this study, the population dynamics of the harmful bloom-forming phytoplankton Heterosigma akashiwo (Raphidophyceae) and the infectious H. akashiwo viruses (HaV) were monitored in Hiroshima Bay, Japan, from May to July 1998. Concurrently, a number of H. akashiwo and HaV clones were isolated, and their virus susceptibilities and host ranges were determined through laboratory cross-reactivity tests. A sudden decrease in cell density of H. akashiwo was accompanied by a drastic increase in the abundance of HaV, suggesting that viruses contributed greatly to the disintegration of the H. akashiwo bloom as mortality agents. Despite the large quantity of infectious HaV, however, a significant proportion of H. akashiwo cells survived after the bloom disintegration. The viral susceptibility of H. akashiwo isolates demonstrated that the majority of these surviving cells were resistant to most of the HaV clones, whereas resistant cells were a minor component during the bloom period. Moreover, these resistant cells were displaced by susceptible cells, presumably due to viral infection. These results demonstrated that the properties of dominant cells within the H. akashiwo population change during the period when a bloom is terminated by viral infection, suggesting that viruses also play an important role in determining the clonal composition and maintaining the clonal diversity of H. akashiwo populations. Therefore, our data indicate that viral infection influences the total abundance and the clonal composition of one host algal species, suggesting that viruses are an important component in quantitatively and qualitatively controlling phytoplankton populations in natural marine environments. PMID:11055943

  17. Immunocapture and microplate-based activity and quantity measurement of pyruvate dehydrogenase in human peripheral blood mononuclear cells.

    PubMed

    Liu, Xiaowen; Pervez, Hira; Andersen, Lars W; Uber, Amy; Montissol, Sophia; Patel, Parth; Donnino, Michael W

    2015-01-01

    Pyruvate dehydrogenase (PDH) activity is altered in many human disorders. Current methods require tissue samples and yield inconsistent results. We describe a modified method for measuring PDH activity from isolated human peripheral blood mononuclear cells (PBMCs). RESULTS/METHODOLOGY: We found that PDH activity and quantity can be successfully measured in human PBMCs. Freeze-thaw cycles cannot efficiently disrupt the mitochondrial membrane. Processing time of up to 20 h does not affect PDH activity with proteinase inhibitor addition and a detergent concentration of 3.3% showed maximum yield. Sample protein concentration is correlated to PDH activity and quantity in human PBMCs from healthy subjects. Measuring PDH activity from PBMCs is a novel, easy and less invasive way to further understand the role of PDH in human disease.

  18. 46 CFR 193.15-5 - Quantity, pipe sizes, and discharge rates.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... VESSELS FIRE PROTECTION EQUIPMENT Carbon Dioxide Extinguishing Systems, Details § 193.15-5 Quantity, pipe sizes, and discharge rates. (a) General. The amount of carbon dioxide required for each space shall be... supply of carbon dioxide need not be provided for each space protected. The total available supply shall...

  19. 46 CFR 95.15-5 - Quantity, pipe sizes, and discharge rates.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... VESSELS FIRE PROTECTION EQUIPMENT Carbon Dioxide Extinguishing Systems, Details § 95.15-5 Quantity, pipe sizes, and discharge rates. (a) General. The amount of carbon dioxide required for each space shall be... supply of carbon dioxide need not be provided for each space protected. The total available supply shall...

  20. 46 CFR 193.15-5 - Quantity, pipe sizes, and discharge rates.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... VESSELS FIRE PROTECTION EQUIPMENT Carbon Dioxide Extinguishing Systems, Details § 193.15-5 Quantity, pipe sizes, and discharge rates. (a) General. The amount of carbon dioxide required for each space shall be... supply of carbon dioxide need not be provided for each space protected. The total available supply shall...

  1. 46 CFR 95.15-5 - Quantity, pipe sizes, and discharge rates.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... VESSELS FIRE PROTECTION EQUIPMENT Carbon Dioxide Extinguishing Systems, Details § 95.15-5 Quantity, pipe sizes, and discharge rates. (a) General. The amount of carbon dioxide required for each space shall be... supply of carbon dioxide need not be provided for each space protected. The total available supply shall...

  2. 46 CFR 95.15-5 - Quantity, pipe sizes, and discharge rates.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... VESSELS FIRE PROTECTION EQUIPMENT Carbon Dioxide Extinguishing Systems, Details § 95.15-5 Quantity, pipe sizes, and discharge rates. (a) General. The amount of carbon dioxide required for each space shall be... supply of carbon dioxide need not be provided for each space protected. The total available supply shall...

  3. 14 CFR § 1216.306 - Actions normally requiring an EIS.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...) § 1216.306 Actions normally requiring an EIS. (a) NASA will prepare an EIS for actions with the potential...) Typical NASA actions normally requiring an EIS include: (1) Development and operation of new launch... using a total quantity of radioactive material greater than the quantity for which the NASA Nuclear...

  4. 14 CFR 139.317 - Aircraft rescue and firefighting: Equipment and agents.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... pounds of potassium-based dry chemical and water with a commensurate quantity of AFFF to total 100 gallons for simultaneous dry chemical and AFFF application. (b) Index B. Either of the following: (1) One... gallons of water and the commensurate quantity of AFFF for foam production. (2) Two vehicles— (i) One...

  5. 14 CFR 139.317 - Aircraft rescue and firefighting: Equipment and agents.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... pounds of potassium-based dry chemical and water with a commensurate quantity of AFFF to total 100 gallons for simultaneous dry chemical and AFFF application. (b) Index B. Either of the following: (1) One... gallons of water and the commensurate quantity of AFFF for foam production. (2) Two vehicles— (i) One...

  6. 14 CFR 139.317 - Aircraft rescue and firefighting: Equipment and agents.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... pounds of potassium-based dry chemical and water with a commensurate quantity of AFFF to total 100 gallons for simultaneous dry chemical and AFFF application. (b) Index B. Either of the following: (1) One... gallons of water and the commensurate quantity of AFFF for foam production. (2) Two vehicles— (i) One...

  7. 14 CFR 139.317 - Aircraft rescue and firefighting: Equipment and agents.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... pounds of potassium-based dry chemical and water with a commensurate quantity of AFFF to total 100 gallons for simultaneous dry chemical and AFFF application. (b) Index B. Either of the following: (1) One... gallons of water and the commensurate quantity of AFFF for foam production. (2) Two vehicles— (i) One...

  8. 14 CFR 139.317 - Aircraft rescue and firefighting: Equipment and agents.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... pounds of potassium-based dry chemical and water with a commensurate quantity of AFFF to total 100 gallons for simultaneous dry chemical and AFFF application. (b) Index B. Either of the following: (1) One... gallons of water and the commensurate quantity of AFFF for foam production. (2) Two vehicles— (i) One...

  9. 29 CFR 1910.109 - Explosives and blasting agents.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... devices. Examples of explosive-actuated power devices are jet tappers and jet perforators. (3) Explosive... more magazines, as a group, must be considered as one magazine, and the total quantity of explosives... located in the same building when one is used only for blasting caps in quantities not in excess of 5,000...

  10. 29 CFR 1910.109 - Explosives and blasting agents.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... devices. Examples of explosive-actuated power devices are jet tappers and jet perforators. (3) Explosive... more magazines, as a group, must be considered as one magazine, and the total quantity of explosives... located in the same building when one is used only for blasting caps in quantities not in excess of 5,000...

  11. 29 CFR 1910.109 - Explosives and blasting agents.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... devices. Examples of explosive-actuated power devices are jet tappers and jet perforators. (3) Explosive... more magazines, as a group, must be considered as one magazine, and the total quantity of explosives... located in the same building when one is used only for blasting caps in quantities not in excess of 5,000...

  12. Macronutrients Association with Change in Waist and Hip Circumference Over 9 Years.

    PubMed

    Lofley, Amy C; Root, Martin M

    2017-01-01

    The calorie distribution of macronutrients affects individuals' health. Quantity and source of macronutrients may play major roles in waist circumference (WC) and hip circumference (HC). This study's purpose is to investigate the association between the quantity and source of macronutrients and the change in WC and HC over 9 years. Participants (N = 11,343) were from the Atherosclerosis Risk in Community Study. Those diagnosed with cancer or a decrease in WC or HC of 15 cm or more over 9 years were excluded. Change scores were created for anthropometrics between clinic visits over 9 years. Macronutrient intakes were assessed by a food frequency questionnaire at visit 1 and presented as a percentage of energy intake. Linear regressions were performed with quartiles of dietary components on change scores for WC and HC with controlling cofactors. Gender subgroup analysis was performed. A larger increase in WC was associated with higher intakes of total carbohydrates, dietary fiber, and fructose (p for trend < 0.005). A smaller increase in WC was associated with higher intakes of sucrose, total protein, animal protein, and alcohol (p for trend < 0.02). A larger increase in HC was associated with higher intakes of total carbohydrate, sucrose, fructose, animal protein, and vegetable fat (p for trend < 0.05). A smaller increase in HC was associated with higher intakes of animal fat, total fat, and total protein (p for trend < 0.05). In males, changes in WC and HC were associated with fructose, sucrose, total fat, and total protein. In females, changes in WC and HC were associated with dietary fiber, fructose, alcohol, animal protein, total protein, animal fat, and vegetable fat. Macronutrient source and quantity play a significant role in individuals' adiposity and effects on WC and HC. Overall, an increase in WC and HC was seen over the 11 years. The source and quantity of the macronutrients play a significant role in WC and HC. Further research needs to be conducted to see the exact effect that macronutrients play on WC and HC.

  13. WT1 isoform expression pattern in acute myeloid leukemia.

    PubMed

    Luna, Irene; Such, Esperanza; Cervera, Jose; Barragán, Eva; Ibañez, Mariam; Gómez-Seguí, Inés; López-Pavía, María; Llop, Marta; Fuster, Oscar; Dolz, Sandra; Oltra, Silvestre; Alonso, Carmen; Vera, Belén; Lorenzo, Ignacio; Martínez-Cuadrón, David; Montesinos, Pau; Senent, M Leonor; Moscardó, Federico; Bolufer, Pascual; Sanz, Miguel A

    2013-12-01

    WT1 plays a dual role in leukemia development, probably due to an imbalance in the expression of the 4 main WT1 isoforms. We quantify their expression and evaluate them in a series of AML patients. Our data showed a predominant expression of isoform D in AML, although in a lower quantity than in normal CD34+ cells. We found a positive correlation between the total WT1 expression and A, B and C isoforms. The overexpression of WT1 in AML might be due to a relative increase in A, B and C isoforms, together with a relative decrease in isoform D expression. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Ionization of biomolecular targets by ion impact: input data for radiobiological applications

    NASA Astrophysics Data System (ADS)

    de Vera, Pablo; Abril, Isabel; Garcia-Molina, Rafael; Solov'yov, Andrey V.

    2013-06-01

    In this work we review and further develop a semiempirical model recently proposed for the ion impact ionization of complex biological media. The model is based on the dielectric formalism, and makes use of a semiempirical parametrization of the optical energy-loss function of bioorganic compounds, allowing the calculation of single and total ionization cross sections and related quantities for condensed biological targets, such as liquid water, DNA and its components, proteins, lipids, carbohydrates or cell constituents. The model shows a very good agreement with experimental data for water, adenine and uracil, and allows the comparison of the ionization efficiency of different biological targets, and also the average kinetic energy of the ejected secondary electrons.

  15. Characterizing regulatory and functional differentiation between maize mesophyll and bundle sheath cells by transcriptomic analysis.

    PubMed

    Chang, Yao-Ming; Liu, Wen-Yu; Shih, Arthur Chun-Chieh; Shen, Meng-Ni; Lu, Chen-Hua; Lu, Mei-Yeh Jade; Yang, Hui-Wen; Wang, Tzi-Yuan; Chen, Sean C-C; Chen, Stella Maris; Li, Wen-Hsiung; Ku, Maurice S B

    2012-09-01

    To study the regulatory and functional differentiation between the mesophyll (M) and bundle sheath (BS) cells of maize (Zea mays), we isolated large quantities of highly homogeneous M and BS cells from newly matured second leaves for transcriptome profiling by RNA sequencing. A total of 52,421 annotated genes with at least one read were found in the two transcriptomes. Defining a gene with more than one read per kilobase per million mapped reads as expressed, we identified 18,482 expressed genes; 14,972 were expressed in M cells, including 53 M-enriched transcription factor (TF) genes, whereas 17,269 were expressed in BS cells, including 214 BS-enriched TF genes. Interestingly, many TF gene families show a conspicuous BS preference in expression. Pathway analyses reveal differentiation between the two cell types in various functional categories, with the M cells playing more important roles in light reaction, protein synthesis and folding, tetrapyrrole synthesis, and RNA binding, while the BS cells specialize in transport, signaling, protein degradation and posttranslational modification, major carbon, hydrogen, and oxygen metabolism, cell division and organization, and development. Genes coding for several transporters involved in the shuttle of C(4) metabolites and BS cell wall development have been identified, to our knowledge, for the first time. This comprehensive data set will be useful for studying M/BS differentiation in regulation and function.

  16. Neuroglian and DE-cadherin activate independent cytoskeleton assembly pathways in Drosophila S2 cells.

    PubMed

    Dubreuil, R R; Grushko, T

    1999-11-19

    The cytoskeletal proteins spectrin and ankyrin colocalize with sites of E-cadherin-mediated cell-cell adhesion in mammalian cells. Here we examined the effects of Drosophila DE-cadherin expression on spectrin and ankyrin in Drosophila S2 tissue culture cells. DE-cadherin caused a dramatic change in the cytoplasmic concentration and distribution of armadillo, the Drosophila homolog of beta catenin. However, DE-cadherin expression had no detectable effect on the quantity or subcellular distribution of ankyrin or spectrin. In reciprocal experiments, recruitment of ankyrin and alphabeta spectrin to the plasma membrane by another cell adhesion molecule, neuroglian, had no effect on the quantity or distribution of armadillo. The results indicate that DE-cadherin-catenin complexes and neuroglian-spectrin/ankyrin complexes form by nonintersecting pathways. Recruitment of spectrin does not appear to be a conserved feature of DE-cadherin function. Copyright 1999 Academic Press.

  17. Metamorphosis of Magnetospirillum magneticum AMB-1 cells

    NASA Astrophysics Data System (ADS)

    Zhang, Fengli; Yu-Zhang, Kui; Zhao, Sanjun; Xiao, Tian; Denis, Michel; Wu, Longfei

    2010-03-01

    Magnetospirillum magneticum strain AMB-1 belongs to the family of magnetotactic bacteria. It possesses a magnetosome chain aligning, with the assistance of cytoskeleton filaments MamK, along the long axis of the spiral cells. Most fresh M. magneticum AMB-1 cells exhibit spiral morphology. In addition, other cell shapes such as curved and spherical were also observed in this organism. Interestingly, the spherical cell shape increased steadily with prolonged incubation time. As the actin-like cytoskeleton protein MreB is involved in maintenance of cell shapes in rod-shaped bacteria such as Escherichia coli and Bacillus subtilis, the correlation between MreB protein levels and cell shape was investigated in this study. Immunoblotting analysis showed that the quantity of MreB decreased when the cell shape changed along with incubation time. As an internal control, the quantity of MamA was not obviously changed under the same conditions. Cell shape directs cell-wall synthesis during growth and division. MreB is required for maintaining the cell shape. Thus, MreB might play an essential role in maintaining the spiral shape of M. magneticum AMB-1 cells.

  18. Expansion and hepatic differentiation of rat multipotent adult progenitor cells in microcarrier suspension culture.

    PubMed

    Park, Y; Subramanian, K; Verfaillie, C M; Hu, W S

    2010-10-01

    Many potential applications of stem cells require large quantities of cells, especially those involving large organs such as the liver. For such applications, a scalable reactor system is desirable to ensure a reliable supply of sufficient quantities of differentiation competent or differentiated cells. We employed a microcarrier culture system for the expansion of undifferentiated rat multipotent adult progenitor cells (rMAPC) as well as for directed differentiation of these cells to hepatocyte-like cells. During the 4-day expansion culture, cell concentration increased by 85-fold while expression level of pluripotency markers were maintained, as well as the MAPC differentiation potential. Directed differentiation into hepatocyte-like cells on the microcarriers themselves gave comparable results as observed with cells cultured in static cultures. The cells expressed several mature hepatocyte-lineage genes and asialoglycoprotein receptor-1 (ASGPR-1) surface protein, and secreted albumin and urea. Microcarrier culture thus offers the potential of large-scale expansion and differentiation of stem cells in a more controlled bioreactor environment. Copyright © 2010 Elsevier B.V. All rights reserved.

  19. Demonstration of landfill gas enhancement techniques in landfill simulators

    NASA Astrophysics Data System (ADS)

    Walsh, J. J.; Vogt, W. G.

    1982-02-01

    Various techniques to enhance gas production in sanitary landfills were applied to landfill simulators. These techniques include (1) accelerated moisture addition, (2) leachate recycling, (3) buffer addition, (4) nutrient addition, and (5) combinations of the above. Results are compiled through on-going operation and monitoring of sixteen landfill simulators. These test cells contain about 380 kg of municipal solid waste. Quantities of buffer and nutrient materials were placed in selected cells at the time of loading. Water is added to all test cells on a monthly basis; leachate is withdrawn from all cells (and recycled on selected cells) also on a monthly basis. Daily monitoring of gas volumes and refuse temperatures is performed. Gas and leachate samples are collected and analyzed on a monthly basis. Leachate and gas quality and quantity reslts are presented for the first 18 months of operation.

  20. Label-free Quantification of Proteins in Single Embryonic Cells with Neural Fate in the Cleavage-Stage Frog (Xenopus laevis) Embryo using Capillary Electrophoresis Electrospray Ionization High-Resolution Mass Spectrometry (CE-ESI-HRMS).

    PubMed

    Lombard-Banek, Camille; Reddy, Sushma; Moody, Sally A; Nemes, Peter

    2016-08-01

    Quantification of protein expression in single cells promises to advance a systems-level understanding of normal development. Using a bottom-up proteomic workflow and multiplexing quantification by tandem mass tags, we recently demonstrated relative quantification between single embryonic cells (blastomeres) in the frog (Xenopus laevis) embryo. In this study, we minimize derivatization steps to enhance analytical sensitivity and use label-free quantification (LFQ) for single Xenopus cells. The technology builds on a custom-designed capillary electrophoresis microflow-electrospray ionization high-resolution mass spectrometry platform and LFQ by MaxLFQ (MaxQuant). By judiciously tailoring performance to peptide separation, ionization, and data-dependent acquisition, we demonstrate an ∼75-amol (∼11 nm) lower limit of detection and quantification for proteins in complex cell digests. The platform enabled the identification of 438 nonredundant protein groups by measuring 16 ng of protein digest, or <0.2% of the total protein contained in a blastomere in the 16-cell embryo. LFQ intensity was validated as a quantitative proxy for protein abundance. Correlation analysis was performed to compare protein quantities between the embryo and n = 3 different single D11 blastomeres, which are fated to develop into the nervous system. A total of 335 nonredundant protein groups were quantified in union between the single D11 cells spanning a 4 log-order concentration range. LFQ and correlation analysis detected expected proteomic differences between the whole embryo and blastomeres, and also found translational differences between individual D11 cells. LFQ on single cells raises exciting possibilities to study gene expression in other cells and models to help better understand cell processes on a systems biology level. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  1. Measurement of beta-amyloid peptides in specific cells using a photo thin-film transistor

    NASA Astrophysics Data System (ADS)

    Kim, Chang-Beom; Chae, Cheol-Joo; Shin, Hye-Rim; Song, Ki-Bong

    2012-01-01

    The existence of beta-amyloid [Aβ] peptides in the brain has been regarded as the most archetypal biomarker of Alzheimer's disease [AD]. Recently, an early clinical diagnosis has been considered a great importance in identifying people who are at high risk of AD. However, no microscale electronic sensing devices for the detection of Aβ peptides have been developed yet. In this study, we propose an effective method to evaluate a small quantity of Aβ peptides labeled with fluorescein isothiocyanate [FITC] using a photosensitive field-effect transistor [p-FET] with an on-chip single-layer optical filter. To accurately evaluate the quantity of Aβ peptides within the cells cultured on the p-FET device, we measured the photocurrents which resulted from the FITC-conjugated Aβ peptides expressed from the cells and measured the number of photons of the fluorochrome in the cells using a photomultiplier tube. Thus, we evaluated the correlation between the generated photocurrents and the number of emitted photons. We also evaluated the correlation between the number of emitted photons and the amount of FITC by measuring the FITC volume using AFM. Finally, we estimated the quantity of Aβ peptides of the cells placed on the p-FET sensing area on the basis of the binding ratio between FITC molecules and Aβ peptides.

  2. Looking inside the box: using Raman microspectroscopy to deconstruct microbial biomass stoichiometry one cell at a time

    USGS Publications Warehouse

    Hall, Edward K.; Singer, Gabriel A.; Pölzl, Marvin; Hämmerle, Ieda; Schwarz, Christian; Daims, Holger; Maixner, Frank; Battin, Tom J.

    2011-01-01

    Stoichiometry of microbial biomass is a key determinant of nutrient recycling in a wide variety of ecosystems. However, little is known about the underlying causes of variance in microbial biomass stoichiometry. This is primarily because of technological constraints limiting the analysis of macromolecular composition to large quantities of microbial biomass. Here, we use Raman microspectroscopy (MS), to analyze the macromolecular composition of single cells of two species of bacteria grown on minimal media over a wide range of resource stoichiometry. We show that macromolecular composition, determined from a subset of identified peaks within the Raman spectra, was consistent with macromolecular composition determined using traditional analytical methods. In addition, macromolecular composition determined by Raman MS correlated with total biomass stoichiometry, indicating that analysis with Raman MS included a large proportion of a cell's total macromolecular composition. Growth phase (logarithmic or stationary), resource stoichiometry and species identity each influenced each organism's macromolecular composition and thus biomass stoichiometry. Interestingly, the least variable peaks in the Raman spectra were those responsible for differentiation between species, suggesting a phylogenetically specific cellular architecture. As Raman MS has been previously shown to be applicable to cells sampled directly from complex environments, our results suggest Raman MS is an extremely useful application for evaluating the biomass stoichiometry of environmental microorganisms. This includes the ability to partition microbial biomass into its constituent macromolecules and increase our understanding of how microorganisms in the environment respond to resource heterogeneity.

  3. Looking inside the box: using Raman microspectroscopy to deconstruct microbial biomass stoichiometry one cell at a time

    PubMed Central

    Hall, Edward K; Singer, Gabriel A; Pölzl, Marvin; Hämmerle, Ieda; Schwarz, Christian; Daims, Holger; Maixner, Frank; Battin, Tom J

    2011-01-01

    Stoichiometry of microbial biomass is a key determinant of nutrient recycling in a wide variety of ecosystems. However, little is known about the underlying causes of variance in microbial biomass stoichiometry. This is primarily because of technological constraints limiting the analysis of macromolecular composition to large quantities of microbial biomass. Here, we use Raman microspectroscopy (MS), to analyze the macromolecular composition of single cells of two species of bacteria grown on minimal media over a wide range of resource stoichiometry. We show that macromolecular composition, determined from a subset of identified peaks within the Raman spectra, was consistent with macromolecular composition determined using traditional analytical methods. In addition, macromolecular composition determined by Raman MS correlated with total biomass stoichiometry, indicating that analysis with Raman MS included a large proportion of a cell's total macromolecular composition. Growth phase (logarithmic or stationary), resource stoichiometry and species identity each influenced each organism's macromolecular composition and thus biomass stoichiometry. Interestingly, the least variable peaks in the Raman spectra were those responsible for differentiation between species, suggesting a phylogenetically specific cellular architecture. As Raman MS has been previously shown to be applicable to cells sampled directly from complex environments, our results suggest Raman MS is an extremely useful application for evaluating the biomass stoichiometry of environmental microorganisms. This includes the ability to partition microbial biomass into its constituent macromolecules and increase our understanding of how microorganisms in the environment respond to resource heterogeneity. PMID:20703314

  4. Mutant quantity and quality in mammalian cells (AL) exposed to cesium-137 gamma radiation: effect of caffeine

    NASA Technical Reports Server (NTRS)

    McGuinness, S. M.; Shibuya, M. L.; Ueno, A. M.; Vannais, D. B.; Waldren, C. A.; Chatterjee, A. (Principal Investigator)

    1995-01-01

    We examined the effect of caffeine (1,3,7-trimethylxanthine) on the quantity and quality of mutations in cultured mammalian AL human-hamster hybrid cells exposed to 137Cs gamma radiation. At a dose (1.5 mg/ml for 16 h) that reduced the plating efficiency (PE) by 20%, caffeine was not itself a significant mutagen, but it increased by approximately twofold the slope of the dose-response curve for induction of S1- mutants by 137Cs gamma radiation. Molecular analysis of 235 S1- mutants using a series of DNA probes mapped to the human chromosome 11 in the AL hybrid cells revealed that 73 to 85% of the mutations in unexposed cells and in cells treated with caffeine alone, 137Cs gamma rays alone or 137Cs gamma rays plus caffeine were large deletions involving millions of base pairs of DNA. Most of these deletions were contiguous with the region of the MIC1 gene at 11p13 that encodes the S1 cell surface antigen. In other mutants that had suffered multiple marker loss, the deletions were intermittent along chromosome 11. These "complex" mutations were rare for 137Cs gamma irradiation (1/63 = 1.5%) but relatively prevalent (23-50%) for other exposure conditions. Thus caffeine appears to alter both the quantity and quality of mutations induced by 137Cs gamma irradiation.

  5. Simultaneous quantification of intracellular and secreted active and inactive glucagon-like peptide-1 from cultured cells.

    PubMed

    Amao, Michiko; Kitahara, Yoshiro; Tokunaga, Ayaka; Shimbo, Kazutaka; Eto, Yuzuru; Yamada, Naoyuki

    2015-03-01

    Glucagon-like peptide-1 (GLP-1) is an incretin peptide that regulates islet hormone secretion. During recent years, incretin-based therapies have been widely used for patients with type 2 diabetes. GLP-1 peptides undergo N- and C-terminal processing for gain or loss of functions. We developed a method to quantify picomolar quantities of intact GLP-1 peptides using liquid chromatography-tandem mass spectrometry (LC-MS/MS). By employing this label-free selected reaction monitoring (SRM) method, we were able to analyze secreted GLP-1(1-37), GLP-1(7-37), and GLP-1(7-36 amid from human enteroendocrine NCI-H716 cells after stimulation with nateglinide, glucose, and sucralose. The absolute total concentrations of secreted GLP-1 peptides at baseline and after stimulation with nateglinide, glucose, and sucralose were 167.3, 498.9, 238.3, and 143.1 pM, respectively. Meanwhile, the ratios of GLP-1(1-37), GLP-1(7-37), and GLP-1(7-36 amide) to total GLP-1 peptides were similar (6 ± 3, 26 ± 3, and 78 ± 5%, respectively). The SRM assay can analyze the concentrations of individual GLP-1 peptides and, therefore, is a tool to investigate the physiological roles of GLP-1 peptides. Furthermore, the molecular species secreted from NCI-H716 cells were unknown. Therefore, we performed a secretopeptidome analysis of supernatants collected from cultured NCI-H716 cells. Together with GLP-1 peptides, we detected neuroendocrine convertase 1, which regulates peptide hormones released from intestinal endocrine L-cells. Copyright © 2014 Elsevier Inc. All rights reserved.

  6. Structures of benthic prokaryotic communities and their hydrolytic enzyme activities resuspended from samples of intertidal mudflats: An experimental approach

    NASA Astrophysics Data System (ADS)

    Mallet, Clarisse; Agogué, Hélène; Bonnemoy, Frédérique; Guizien, Katell; Orvain, Francis; Dupuy, Christine

    2014-09-01

    Resuspended sediment can increase plankton biomass and the growth of bacteria, thus influencing the coastal planktonic microbial food web. But little is known about resuspension itself: is it a single massive change or a whole series of events and how does it affect the quantity and quality of resuspended prokaryotic cells? We simulated the sequential erosion of mud cores to better understand the fate and role of benthic prokaryotes resuspended in the water column. We analyzed the total, attached and free-living prokaryotic cells resuspended, their structure and the activities of their hydrolytic enzymes in terms of the biotic and abiotic factors that affect the composition of microphytobenthic biofilm. Free living prokaryotes were resuspended during the fluff layer erosion phase (for shear velocities below 5 cm · s- 1) regardless of the bed sediment composition. At the higher shear velocities, resuspended prokaryotes were attached to particulate matter. Free and attached cells are thus unevenly distributed, scattered throughout the organic matter (OM) in the uppermost mm of the sediment. Only 10-27% of the total cells initially resuspended were living and most of the Bacteria were Cyanobacteria and Gamma-proteobacteria; their numbers increased to over 30% in parallel with the hydrolytic enzyme activity at highest shear velocity. These conditions released prokaryotic cells having different functions that lie deep in the sediment; the most important of them are Archaea. Finally, composition of resuspended bacterial populations varied with resuspension intensity, and intense resuspension events boosted the microbial dynamics and enzyme activities in the bottom layers of sea water.

  7. Protein deposition on a lathe-cut silicone hydrogel contact lens material.

    PubMed

    Subbaraman, Lakshman N; Woods, Jill; Teichroeb, Jonathan H; Jones, Lyndon

    2009-03-01

    To determine the quantity of total protein, total lysozyme, and the conformational state of lysozyme deposited on a novel, lathe-cut silicone hydrogel (SiHy) contact lens material (sifilcon A) after 3 months of wear. Twenty-four subjects completed a prospective, bilateral, daily-wear, 9-month clinical evaluation in which the subjects were fitted with a novel, custom-made, lathe-cut SiHy lens material. The lenses were worn for three consecutive 3-month periods, with lenses being replaced after each period of wear. After 3 months of wear, the lenses from the left eye were collected and assessed for protein analysis. The total protein deposited on the lenses was determined by a modified Bradford assay, total lysozyme using Western blotting and the lysozyme activity was determined using a modified micrococcal assay. The total protein recovered from the custom-made lenses was 5.3 +/- 2.3 microg/lens and the total lysozyme was 2.4 +/- 1.2 microg/lens. The denatured lysozyme found on the lenses was 1.9 +/- 1.0 microg/lens and the percentage of lysozyme denatured was 80 +/- 10%. Even after 3 months of wear, the quantity of protein and the conformational state of lysozyme deposited on these novel lens materials was very similar to that found on similar surface-coated SiHy lenses after 2 to 4 weeks of wear. These results indicate that extended use of the sifilcon A material is not deleterious in terms of the quantity and quality of protein deposited on the lens.

  8. [Construction of rAAV2-GPIIb/IIIa vector and test of its expression and function in vitro].

    PubMed

    Wang, Kai; Peng, Jian-Qiang; Chen, Fang-Ping; Wu, Xiao-Bin

    2006-04-01

    This study was aimed to explore the possibility of rAAV2 vector-mediating gene therapy for Glanzmann' s thrombasthenia. The rAAV2-GPIIb/IIIa vector was constructed. The GPIIb/IIIa gene expression in mammal cell were examined by different methods, such as: detection of mRNA expression in BHK-21 cells after 24 hours of infection (MOI = 1 x 10(5) v.g/cell) was performed by RT-PCR; the relation between MOI and quantity of GPII6/IIIa gene expression was detected by FACS after 48 hours of infection; GPIIb/IIIa protein expression in BHK-21 cells after 48 hours of infection (MOI = 10(5) v x g/cell) was assayed by Western blot, GPIIb/IIIa protein expression on cell surface was detected by immunofluorescence, and the biological function of expressing product was determined by PAC-1 conjunct experiments. The results showed that GPIIb/IIIa gene expression in mRNA level could be detected in BHK-21 cells after 24 hours of infection at MOI = 1 x 10(5) v x g/cell and the GPIIb/IIIa gene expression in protein level could be detected in BHK-21 cells after 48 hours of infection at MOI = 1 x 10(5) v x g/cell. In certain range, quantity of GPIIb/IIIa gene expression increased with MOI, but overdose of MOI decreased quantity of GPIIb/IIIa gene expression. Activated product of GPIIb/IIIa gene expression could combined with PAC-I, and possesed normal biological function. In conclusion, rAAV2 vactor can effectively mediate GPIIb and GPIIIa gene expressing in mammal cells, and the products of these genes exhibit biological function. This result may provide a basis for application of rAAV2 vector in Glanzmann's thrombasthenia gene therapy in furture.

  9. Changes Are Needed to the Army Contract with Sikorsky to Use Existing DoD Inventory and Control Costs at the Corpus Christi Army Depot

    DTIC Science & Technology

    2011-11-03

    contingency stock for requirements outside the CCAD/Sikorsky contract. 6 The DoD EMALL is a full-service eCommerce site, which strives to be the single...such quantity as (A) will result in the total cost and unit cost most advantageous to the United States where practicable.” Having more than one...practice and frequently does not allow DoD to take advantage of economic order quantities. Table 15 shows examples where higher quantities procured

  10. Characteristics and in vitro Anti-diabetic Properties of the Korean Rice Wine, Makgeolli Fermented with Laminaria japonica

    PubMed Central

    Choi, Jae-Suk; Seo, Hyo Ju; Lee, Yu-Ri; Kwon, Su-Jung; Moon, Sun Hwa; Park, Sun-Mee; Sohn, Jae Hak

    2014-01-01

    New in vitro anti-diabetes makgeolli was produced from rice by adding various quantities of Laminaria japonica, and the fermentation characteristics of the L. japonica makgeolli during the fermentation process were investigated. The contents of alcohol and reducing sugar, and viable count of yeast, of L. japonica makgeolli were not significantly changed when the proportion of L. japonica was increased. The total acid content decreased with an increase in L. japonica concentration; the pH and total bacterial cell count increased in proportion with the increase in L. japonica concentration. The L. japonica makgeolli contents of free sugars, such as fructose, glucose, and sucrose, and of organic acids, such as acetic acid, citric acid, succinic acid, and lactic acid, were altered during fermentation and showed various patterns. The effects of the quantity of L. japonica added on the acceptability and anti-diabetes activities of L. japonica makgeolli were also investigated. In a sensory evaluation, L. japonica makgeolli brewed by adding 2.5 or 5% L. japonica to the mash showed the best overall acceptability; the 12.5% L. japonica sample was least favored due to its seaweed flavor. L. japonica addition did not increase the peroxynitrite-scavenging activity of makgeolli. L. japonica makgeolli showed potent anti-diabetes activity, particularly that containing >7.5% L. japonica. Therefore, L. japonica makgeolli may represent a new functional makgeolli with anti-diabetes properties. PMID:25054108

  11. Islet product characteristics and factors related to successful human islet transplantation from the Collaborative Islet Transplant Registry (CITR) 1999-2010.

    PubMed

    Balamurugan, A N; Naziruddin, B; Lockridge, A; Tiwari, M; Loganathan, G; Takita, M; Matsumoto, S; Papas, K; Trieger, M; Rainis, H; Kin, T; Kay, T W; Wease, S; Messinger, S; Ricordi, C; Alejandro, R; Markmann, J; Kerr-Conti, J; Rickels, M R; Liu, C; Zhang, X; Witkowski, P; Posselt, A; Maffi, P; Secchi, A; Berney, T; O'Connell, P J; Hering, B J; Barton, F B

    2014-11-01

    The Collaborative Islet Transplant Registry (CITR) collects data on clinical islet isolations and transplants. This retrospective report analyzed 1017 islet isolation procedures performed for 537 recipients of allogeneic clinical islet transplantation in 1999-2010. This study describes changes in donor and islet isolation variables by era and factors associated with quantity and quality of final islet products. Donor body weight and BMI increased significantly over the period (p<0.001). Islet yield measures have improved with time including islet equivalent (IEQ)/particle ratio and IEQs infused. The average dose of islets infused significantly increased in the era of 2007-2010 when compared to 1999-2002 (445.4±156.8 vs. 421.3±155.4×0(3) IEQ; p<0.05). Islet purity and total number of β cells significantly improved over the study period (p<0.01 and <0.05, respectively). Otherwise, the quality of clinical islets has remained consistently very high through this period, and differs substantially from nonclinical islets. In multivariate analysis of all recipient, donor and islet factors, and medical management factors, the only islet product characteristic that correlated with clinical outcomes was total IEQs infused. This analysis shows improvements in both quantity and some quality criteria of clinical islets produced over 1999-2010, and these parallel improvements in clinical outcomes over the same period. © 2014 The Authors. American Journal of Transplantation Published by Wiley Periodicals, Inc. on behalf of American Society of Transplant Surgeons.

  12. Islet Product Characteristics and Factors Related to Successful Human Islet Transplantation From the Collaborative Islet Transplant Registry (CITR) 1999–2010

    PubMed Central

    Balamurugan, A N; Naziruddin, B; Lockridge, A; Tiwari, M; Loganathan, G; Takita, M; Matsumoto, S; Papas, K; Trieger, M; Rainis, H; Kin, T; Kay, T W; Wease, S; Messinger, S; Ricordi, C; Alejandro, R; Markmann, J; Kerr-Conti, J; Rickels, M R; Liu, C; Zhang, X; Witkowski, P; Posselt, A; Maffi, P; Secchi, A; Berney, T; O’Connell, P J; Hering, B J; Barton, F B

    2014-01-01

    The Collaborative Islet Transplant Registry (CITR) collects data on clinical islet isolations and transplants. This retrospective report analyzed 1017 islet isolation procedures performed for 537 recipients of allogeneic clinical islet transplantation in 1999–2010. This study describes changes in donor and islet isolation variables by era and factors associated with quantity and quality of final islet products. Donor body weight and BMI increased significantly over the period (p < 0.001). Islet yield measures have improved with time including islet equivalent (IEQ)/particle ratio and IEQs infused. The average dose of islets infused significantly increased in the era of 2007–2010 when compared to 1999–2002 (445.4 ± 156.8 vs. 421.3 ± 155.4 ×103 IEQ; p < 0.05). Islet purity and total number of β cells significantly improved over the study period (p < 0.01 and <0.05, respectively). Otherwise, the quality of clinical islets has remained consistently very high through this period, and differs substantially from nonclinical islets. In multivariate analysis of all recipient, donor and islet factors, and medical management factors, the only islet product characteristic that correlated with clinical outcomes was total IEQs infused. This analysis shows improvements in both quantity and some quality criteria of clinical islets produced over 1999–2010, and these parallel improvements in clinical outcomes over the same period. PMID:25278159

  13. The response of belowground carbon allocation in forests to global change

    Treesearch

    Christian P. Giardina; Mark Coleman; Dan Binkley; Jessica Hancock; John S. King; Erik Lilleskov; Wendy M. Loya; Kurt S. Pregitzer; Michael G. Ryan; Carl Trettin

    2005-01-01

    Belowground carbon allocation (BCA) in forests regulates soil organic matter formation and influences biotic and abiotic properties of soil such as bulk density, cation exchange capacity, and water holding capacity. On a global scale, the total quantity of carbon allocated belowground by terrestrial plants is enormous, exceeding by an order of magnitude the quantity of...

  14. 29 CFR 1910.109 - Explosives and blasting agents.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... more magazines, as a group, must be considered as one magazine, and the total quantity of explosives... located in the same building when one is used only for blasting caps in quantities not in excess of 5,000... distance of at least one hundred and fifty (150) feet shall be maintained between class II magazines and...

  15. [Using Excess Activated Sludge Treated 4-Chlorophenol Contained Waste Water to Cultivate Chlorella vulgaris].

    PubMed

    Wang, Lu; Chen, Xiu-rong; Yan, Long; He, Yi-xuan; Shi, Zhen-dong

    2015-04-01

    Using different rations of sludge extracts and supernate from 4-Chlorophenol (4-CP) simulated wastewater's excess sludge after centrifugation to cultivate the Chlorella vulgaris to achieve the goal of excess sludge utilization together with chlorella cultivating. The experiments were performed in 500 mL flasks with different rations of sludge extracts & BG-11 and supernate & BG-11 in a light growth chamber respectively. Number of algal cells, Chlorophyll, enzyme activity, oil and water total nitrogen (TN), total phosphorus (TP), total organic carbon (TOC), toxicity index were investigated. Result showed that the nutrition supplies and toxicity in the excess sludge were removed efficiently via Chlorella vulgaris, the removal rates of TN and TP were at least 40% and 90% respectively; After 10 days cultivation, the density growth of 50% sludge extracts was 20 times higher of the beginning while its chlorophyll content was lower than that of the blank group. Sludge extracts could promote the proliferation of algae, but were not conducive to the synthesis of chlorophyll. The quantity of SOD in per cell showed Chlorella vulgaris gave a positive response via stimulation from toxicant in sludge extracts and supernate. The best time for collecting chlorella vulgaris was the fifth day of cultivation, taking neutral oil accumulation as the evaluating indicator for its utilization combined with the removal of supplies and toxicity.

  16. Naive T-cell receptor transgenic T cells help memory B cells produce antibody

    PubMed Central

    Duffy, Darragh; Yang, Chun-Ping; Heath, Andrew; Garside, Paul; Bell, Eric B

    2006-01-01

    Injection of the same antigen following primary immunization induces a classic secondary response characterized by a large quantity of high-affinity antibody of an immunoglobulin G class produced more rapidly than in the initial response – the products of memory B cells are qualitatively distinct from that of the original naive B lymphocytes. Very little is known of the help provided by the CD4 T cells that stimulate memory B cells. Using antigen-specific T-cell receptor transgenic CD4 T cells (DO11.10) as a source of help, we found that naive transgenic T cells stimulated memory B cells almost as well (in terms of quantity and speed) as transgenic T cells that had been recently primed. There was a direct correlation between serum antibody levels and the number of naive transgenic T cells transferred. Using T cells from transgenic interleukin-2-deficient mice we showed that interleukin-2 was not required for a secondary response, although it was necessary for a primary response. The results suggested that the signals delivered by CD4 T cells and required by memory B cells for their activation were common to both antigen-primed and naive CD4 T cells. PMID:17067314

  17. Method for operating a combustor in a fuel cell system

    DOEpatents

    Chalfant, Robert W.; Clingerman, Bruce J.

    2002-01-01

    A method of operating a combustor to heat a fuel processor in a fuel cell system, in which the fuel processor generates a hydrogen-rich stream a portion of which is consumed in a fuel cell stack and a portion of which is discharged from the fuel cell stack and supplied to the combustor, and wherein first and second streams are supplied to the combustor, the first stream being a hydrocarbon fuel stream and the second stream consisting of said hydrogen-rich stream, the method comprising the steps of monitoring the temperature of the fuel processor; regulating the quantity of the first stream to the combustor according to the temperature of the fuel processor; and comparing said quantity of said first stream to a predetermined value or range of predetermined values.

  18. Immunocapture and microplate-based activity and quantity measurement of pyruvate dehydrogenase in human peripheral blood mononuclear cells

    PubMed Central

    Liu, Xiaowen; Pervez, Hira; Andersen, Lars W; Uber, Amy; Montissol, Sophia; Patel, Parth; Donnino, Michael W

    2015-01-01

    Background Pyruvate dehydrogenase (PDH) activity is altered in many human disorders. Current methods require tissue samples and yield inconsistent results. We describe a modified method for measuring PDH activity from isolated human peripheral blood mononuclear cells (PBMCs). Results/Methodology We found that PDH activity and quantity can be successfully measured in human PBMCs. Freeze-thaw cycles cannot efficiently disrupt the mitochondrial membrane. Processing time of up to 20 h does not affect PDH activity with proteinase inhibitor addition and a detergent concentration of 3.3% showed maximum yield. Sample protein concentration is correlated to PDH activity and quantity in human PBMCs from healthy subjects. Conclusion Measuring PDH activity from PBMCs is a novel, easy and less invasive way to further understand the role of PDH in human disease. PMID:25826140

  19. Myocardial Autophagy after Severe Burn in Rats

    PubMed Central

    Zhang, Qiong; Shi, Xiao-hua; Huang, Yue-sheng

    2012-01-01

    Background Autophagy plays a major role in myocardial ischemia and hypoxia injury. The present study investigated the effects of autophagy on cardiac dysfunction in rats after severe burn. Methods Protein expression of the autophagy markers LC3 and Beclin 1 were determined at 0, 1, 3, 6, and 12 h post-burn in Sprague Dawley rats subjected to 30% total body surface area 3rd degree burns. Autophagic, apoptotic, and oncotic cell death were evaluated in the myocardium at each time point by immunofluorescence. Changes of cardiac function were measured in a Langendorff model of isolated heart at 6 h post-burn, and the autophagic response was measured following activation by Rapamycin and inhibition by 3-methyladenine (3-MA). The angiotensin converting enzyme inhibitor enalaprilat, the angiotensin receptor I blocker losartan, and the reactive oxygen species inhibitor diphenylene iodonium (DPI) were also applied to the ex vivo heart model to examine the roles of these factors in post-burn cardiac function. Results Autophagic cell death was first observed in the myocardium at 3 h post-burn, occurring in 0.008 ± 0.001% of total cardiomyocytes, and continued to increase to a level of 0.022 ± 0.005% by 12 h post-burn. No autophagic cell death was observed in control hearts. Compared with apoptosis, autophagic cell death occurred earlier and in larger quantities. Rapamycin enhanced autophagy and decreased cardiac function in isolated hearts 6 h post-burn, while 3-MA exerted the opposite response. Enalaprilat, losartan, and DPI all inhibited autophagy and enhanced heart function. Conclusion Myocardial autophagy is enhanced in severe burns and autophagic cell death occurred early at 3 h post-burn, which may contribute to post-burn cardiac dysfunction. Angiotensin II and reactive oxygen species may play important roles in this process by regulating cell signaling transduction. PMID:22768082

  20. Multi-Scale Computational Modeling of Two-Phased Metal Using GMC Method

    NASA Technical Reports Server (NTRS)

    Moghaddam, Masoud Ghorbani; Achuthan, A.; Bednacyk, B. A.; Arnold, S. M.; Pineda, E. J.

    2014-01-01

    A multi-scale computational model for determining plastic behavior in two-phased CMSX-4 Ni-based superalloys is developed on a finite element analysis (FEA) framework employing crystal plasticity constitutive model that can capture the microstructural scale stress field. The generalized method of cells (GMC) micromechanics model is used for homogenizing the local field quantities. At first, GMC as stand-alone is validated by analyzing a repeating unit cell (RUC) as a two-phased sample with 72.9% volume fraction of gamma'-precipitate in the gamma-matrix phase and comparing the results with those predicted by finite element analysis (FEA) models incorporating the same crystal plasticity constitutive model. The global stress-strain behavior and the local field quantity distributions predicted by GMC demonstrated good agreement with FEA. High computational saving, at the expense of some accuracy in the components of local tensor field quantities, was obtained with GMC. Finally, the capability of the developed multi-scale model linking FEA and GMC to solve real life sized structures is demonstrated by analyzing an engine disc component and determining the microstructural scale details of the field quantities.

  1. Counter-flow elutriation of clinical peripheral blood mononuclear cell concentrates for the production of dendritic and T cell therapies.

    PubMed

    Stroncek, David F; Fellowes, Vicki; Pham, Chauha; Khuu, Hanh; Fowler, Daniel H; Wood, Lauren V; Sabatino, Marianna

    2014-09-17

    Peripheral blood mononuclear cells (PBMC) concentrates collected by apheresis are frequently used as starting material for cellular therapies, but the cell of interest must often be isolated prior to initiating manufacturing. The results of enriching 59 clinical PBMC concentrates for monocytes or lymphocytes from patients with solid tumors or multiple myeloma using a commercial closed system semi-automated counter-flow elutriation instrument (Elutra, Terumo BCT) were evaluated for quality and consistency. Elutriated monocytes (n = 35) were used to manufacture autologous dendritic cells and elutriated lymphocytes (n = 24) were used manufacture autologous T cell therapies. Elutriated monocytes with >10% neutrophils were subjected to density gradient sedimentation to reduce neutrophil contamination and elutriated lymphocytes to RBC lysis. Elutriation separated the PBMC concentrates into 5 fractions. Almost all of the lymphocytes, platelets and red cells were found in fractions 1 and 2; in contrast, most of the monocytes, 88.6 ± 43.0%, and neutrophils, 74.8 ± 64.3%, were in fraction 5. In addition, elutriation of 6 PBMCs resulted in relatively large quantities of monocytes in fractions 1 or 2. These 6 PBMCs contained greater quantities of monocytes than the other 53 PBMCs. Among fraction 5 isolates 38 of 59 contained >10% neutrophils. High neutrophil content of fraction 5 was associated with greater quantities of neutrophils in the PBMC concentrate. Following density gradient separation the neutrophil counts fell to 3.6 ± 3.4% (all products contained <10% neutrophils). Following red cell lysis of the elutriated lymphocyte fraction the lymphocyte recovery was 86.7 ± 24.0% and 34.3 ± 37.4% of red blood cells remained. Elutriation was consistent and effective for isolating monocytes and lymphocytes from PBMC concentrates for manufacturing clinical cell therapies, but further processing is often required.

  2. Effects of weightlessness on body composition in the rat

    NASA Technical Reports Server (NTRS)

    Pitts, G. C.; Ushakov, A. S.; Pace, N.; Smith, A. H.; Rahlmann, D. F.; Smirnova, T. A.

    1983-01-01

    The effects of weightlessness on the body composition of rats were investigated using 5 male rats exposed to 18.5 days of weightlessness on the COSMOS 1129 biosatellite and killed after reentry. The animals were immediately dissected and the three major body divisions (musculoskeletal system, skin, and pooled viscera) were analyzed for fat, water, solids, and six elements. These results were determined as percentages of the fat-free body or its components and then compared with two groups of terrestrial controls, one of which was subjected to a flight simulation in a spacecraft mock-up while the other was under standard vivarium conditions. Compared with the control groups, the flight group was found to exhibit a reduced fraction of total body water, a net shift of body water from skin to viscera, a marked diminution in the fraction of extracellular water in the fat-free body, a marked reduction in the fraction of bone mineral, no change in the quantity of stored fat or adrenal masses, and a net increase in total muscle mass as indicated by total body creatine, protein, and body cell mass.

  3. Scenedesmus sp. NJ-1 isolated from Antarctica: a suitable renewable lipid source for biodiesel production.

    PubMed

    Chen, Zhuo; Gong, Yangmin; Fang, Xiantao; Hu, Hanhua

    2012-11-01

    Microalgal lipids are promising alternative feedstocks for biodiesel production. Scenedesmus sp. NJ-1, an oil-rich freshwater microalga isolated from Antarctica, was identified to be a suitable candidate to produce biodiesel in this study. This strain could grow at temperatures ranging from 4 to 35 °C. With regular decrease in nitrate concentration in the medium, large quantities of triacylglycerols accumulated under batch culture conditions detected by thin layer chromatography and BODIPY 505/515 fluorescent staining. Scenedesmus sp. NJ-1 achieved the average biomass productivity of 0.105 g l⁻¹ d⁻¹ (dry weight) and nearly the highest lipid content (35 % of dry cell weight) was reached at day 28 in the batch culture. Neutral lipids accounted for 78 % of total lipids, and C18:1 (n-9), C16:0 were the major fatty acids in total lipids, composing 37 and 20 % of total fatty acids of Scenedesmus sp. NJ-1 grown for 36 days, respectively. These results suggested that Scenedesmus sp. NJ-1 was a good source of microalgal oils for biodiesel production.

  4. Explicit tracking of uncertainty increases the power of quantitative rule-of-thumb reasoning in cell biology.

    PubMed

    Johnston, Iain G; Rickett, Benjamin C; Jones, Nick S

    2014-12-02

    Back-of-the-envelope or rule-of-thumb calculations involving rough estimates of quantities play a central scientific role in developing intuition about the structure and behavior of physical systems, for example in so-called Fermi problems in the physical sciences. Such calculations can be used to powerfully and quantitatively reason about biological systems, particularly at the interface between physics and biology. However, substantial uncertainties are often associated with values in cell biology, and performing calculations without taking this uncertainty into account may limit the extent to which results can be interpreted for a given problem. We present a means to facilitate such calculations where uncertainties are explicitly tracked through the line of reasoning, and introduce a probabilistic calculator called CALADIS, a free web tool, designed to perform this tracking. This approach allows users to perform more statistically robust calculations in cell biology despite having uncertain values, and to identify which quantities need to be measured more precisely to make confident statements, facilitating efficient experimental design. We illustrate the use of our tool for tracking uncertainty in several example biological calculations, showing that the results yield powerful and interpretable statistics on the quantities of interest. We also demonstrate that the outcomes of calculations may differ from point estimates when uncertainty is accurately tracked. An integral link between CALADIS and the BioNumbers repository of biological quantities further facilitates the straightforward location, selection, and use of a wealth of experimental data in cell biological calculations. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  5. Residual monomers and degree of conversion of partially bioresorbable fiber-reinforced composite.

    PubMed

    Väkiparta, Marju; Puska, Mervi; Vallittu, Pekka K

    2006-01-01

    The aim of this study was to evaluate the total quantity of residual monomer (bis-phenyl glycidyl dimethacrylate, i.e. Bis-GMA, and triethylene glycol dimethacrylate, i.e. TEGDMA), residual monomer release into water and the degree of monomer conversion (DC%) of glass fiber-reinforced composites (FRC) with a partially bioresorbable polymer matrix. Another aim was to find out whether the curing mode affects the quantity of residual monomer and degree of conversion. Glass fibers were preimpregnated with a bioresorbable poly(hydroxyproline) amide and non-resorbable Bis-GMA-TEGDMA resin system. Specimens were immersed in water for 1, 3 or 7 days (37 degrees C) to determine the quantity of leached residual monomers, or in the solvent tetrahydrofuran for 3 days to determine the total quantity of residual monomers by high performance liquid chromatography. DC% was measured by Fourier transform infrared spectroscopy. The quantity of residual monomer of the specimens decreased when the specimens contained glass fibers, and/or poly(hydroxyproline) amide, and/or when it was post-cured. The majority of the residual monomers were leached out during the first 24 h of immersion in water. The DC% of the specimens increased when post-cured. Also glass fibers in the composite increased the DC% in contrast to Bis-GMA-TEGDMA resin only. In conclusion, use of poly(hydroxyproline) amide as a sizing of the glass fibers in FRC does not increase the quantity of residual monomers. These results suggest that this new kind of partially bioresorbable FRC has potential for biomedical applications.

  6. Exercise quantity-dependent muscle hypertrophy in adult zebrafish (Danio rerio).

    PubMed

    Hasumura, Takahiro; Meguro, Shinichi

    2016-07-01

    Exercise is very important for maintaining and increasing skeletal muscle mass, and is particularly important to prevent and care for sarcopenia and muscle disuse atrophy. However, the dose-response relationship between exercise quantity, duration/day, and overall duration and muscle mass is poorly understood. Therefore, we investigated the effect of exercise duration on skeletal muscle to reveal the relationship between exercise quantity and muscle hypertrophy in zebrafish forced to exercise. Adult male zebrafish were exercised 6 h/day for 4 weeks, 6 h/day for 2 weeks, or 3 h/day for 2 weeks. Flow velocity was adjusted to maximum velocity during continual swimming (initial 43 cm/s). High-speed consecutive photographs revealed that zebrafish mainly drove the caudal part. Additionally, X-ray micro computed tomography measurements indicated muscle hypertrophy of the mid-caudal half compared with the mid-cranial half part. The cross-sectional analysis of the mid-caudal half muscle revealed that skeletal muscle (red, white, or total) mass increased with increasing exercise quantity, whereas that of white muscle and total muscle increased only under the maximum exercise load condition of 6 h/day for 4 weeks. Additionally, the muscle fiver size distributions of exercised fish were larger than those from non-exercised fish. We revealed that exercise quantity, duration/day, and overall duration were correlated with skeletal muscle hypertrophy. The forced exercise model enabled us to investigate the relationship between exercise quantity and skeletal muscle mass. These results open up the possibility for further investigations on the effects of exercise on skeletal muscle in adult zebrafish.

  7. Enhancing Least-Squares Finite Element Methods Through a Quantity-of-Interest

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chaudhry, Jehanzeb Hameed; Cyr, Eric C.; Liu, Kuo

    2014-12-18

    Here, we introduce an approach that augments least-squares finite element formulations with user-specified quantities-of-interest. The method incorporates the quantity-of-interest into the least-squares functional and inherits the global approximation properties of the standard formulation as well as increased resolution of the quantity-of-interest. We establish theoretical properties such as optimality and enhanced convergence under a set of general assumptions. Central to the approach is that it offers an element-level estimate of the error in the quantity-of-interest. As a result, we introduce an adaptive approach that yields efficient, adaptively refined approximations. Several numerical experiments for a range of situations are presented to supportmore » the theory and highlight the effectiveness of our methodology. Notably, the results show that the new approach is effective at improving the accuracy per total computational cost.« less

  8. Detection of Sendai virus receptor, the ganglioside GDla, in target tissue (mouse lung)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Markwell, M.A.K.; Sato, E.

    1986-05-01

    Previously the authors had shown that the gangliosides GDla, GTlb, and GQlb derived from brain function as receptors for the paramyxovirus Sendai virus by their ability to induce infection when incubated with receptor-deficient cells. Analyses of MDBK, HeLa, and MDCK cells in culture demonstrated that these putative receptors were present in host cells in the quantities required for infection. The primary site of infection for Sendai virus in the whole animal is the respiratory tract, culminating in the lung. Therefore, the ganglioside content of this target organ was analyzed to determine the endogenous receptor population available to Sendai virus. Themore » total ganglioside fraction of lung was resolved into individual species by HPTLC. Gangliosides of the gangliotetraose series were identified by the specific binding of /sup 125/I-labeled tetanus and cholera toxins before and after exposure with sialidase. In this manner one of the major resorcinol-positive bands was identified as GDla. Evidence of the more complex ganglioside receptors for Sendai virus was also seen.« less

  9. Immobilization of Actinobacillus succinogenes by adhesion or entrapment for the production of succinic acid.

    PubMed

    Corona-González, Rosa Isela; Miramontes-Murillo, Ricardo; Arriola-Guevara, Enrique; Guatemala-Morales, Guadalupe; Toriz, Guillermo; Pelayo-Ortiz, Carlos

    2014-07-01

    The production of succinic acid was studied with entrapped and adsorbed Actinobacillus succinogenes. The adsorption of fermentation products (organic acids in the concentration range of 1-20 g/L) on different supports was evaluated. It was found that succinic acid was adsorbed in small quantities on diatomite and zeolite (12.6 mg/g support). The highest production of succinic acid was achieved with A. succinogenes entrapped in agar beads. Batch fermentations with immobilized cells were carried out with glucose concentrations ranging from 20 to 80 g/L. Succinic acid (43.4 g/L) was obtained from 78.3g/L glucose, and a high productivity (2.83 g/Lh) was obtained with a glucose concentration of 37.6g/L. For repeated batch fermentations (5 cycles in 72 h) with immobilized cells in agar, the total glucose consumed was 147.55 g/L, while the production of succinic acid was 107 g/L. Immobilized cells reduced significantly the fermentation time, yield, productivity and final concentration of succinic acid. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Cyanobacteria and Cyanotoxins Occurrence and Removal from Five High-Risk Conventional Treatment Drinking Water Plants.

    PubMed

    Szlag, David C; Sinclair, James L; Southwell, Benjamin; Westrick, Judy A

    2015-06-12

    An environmental protection agency EPA expert workshop prioritized three cyanotoxins, microcystins, anatoxin-a, and cylindrospermopsin (MAC), as being important in freshwaters of the United States. This study evaluated the prevalence of potentially toxin producing cyanobacteria cell numbers relative to the presence and quantity of the MAC toxins in the context of this framework. Total and potential toxin producing cyanobacteria cell counts were conducted on weekly raw and finished water samples from utilities located in five US states. An Enzyme-Linked Immunosorbant Assay (ELISA) was used to screen the raw and finished water samples for microcystins. High-pressure liquid chromatography with a photodiode array detector (HPLC/PDA) verified microcystin concentrations and quantified anatoxin-a and cylindrospermopsin concentrations. Four of the five utilities experienced cyanobacterial blooms in their raw water. Raw water samples from three utilities showed detectable levels of microcystins and a fourth utility had detectable levels of both microcystin and cylindrospermopsin. No utilities had detectable concentrations of anatoxin-a. These conventional plants effectively removed the cyanobacterial cells and all finished water samples showed MAC levels below the detection limit by ELISA and HPLC/PDA.

  11. Low-dose lisinopril in normotensive men with idiopathic oligospermia and infertility: a 5-year randomized, controlled, crossover pilot study.

    PubMed

    Mbah, A U; Ndukwu, G O; Ghasi, S I; Shu, E N; Ozoemena, F N; Mbah, J O; Onodugo, O D; Ejim, E C; Eze, M I; Nkwo, P O; Okonkwo, P O

    2012-04-01

    The outcomes of drug treatment for male infertility remain conjectural, with controversial study results. Our pilot study employed a randomized, placebo-controlled, crossover methodology with intention-to-treat analysis. Thirty-three men with idiopathic oligospermia were randomized to start either daily oral lisinopril 2.5 mg (n = 17) or daily oral placebo (n = 16). Lisinopril was found to cause a normalization of seminal parameters in 53.6% of the participants. Although the mean ejaculate volume was unchanged (P ≥ 0.093), the total sperm cell count and the percentage of motile sperm cells increased (P ≤ 0.03 and P < 0.001, respectively), whereas the percentage of sperm cells with abnormal morphology decreased (P ≤ 0.04). The pregnancy rate was 48.5%, and there was no serious adverse drug event. It is concluded, albeit cautiously, that prolonged treatment with 2.5 mg/day of oral lisinopril may be well tolerated in normotensive men with idiopathic oligospermia, may improve sperm quantity and quality, and may enhance fertility in approximately half of those treated.

  12. A Cell Wall Proteome and Targeted Cell Wall Analyses Provide Novel Information on Hemicellulose Metabolism in Flax.

    PubMed

    Chabi, Malika; Goulas, Estelle; Leclercq, Celine C; de Waele, Isabelle; Rihouey, Christophe; Cenci, Ugo; Day, Arnaud; Blervacq, Anne-Sophie; Neutelings, Godfrey; Duponchel, Ludovic; Lerouge, Patrice; Hausman, Jean-François; Renaut, Jenny; Hawkins, Simon

    2017-09-01

    Experimentally-generated (nanoLC-MS/MS) proteomic analyses of four different flax organs/tissues (inner-stem, outer-stem, leaves and roots) enriched in proteins from 3 different sub-compartments (soluble-, membrane-, and cell wall-proteins) was combined with publically available data on flax seed and whole-stem proteins to generate a flax protein database containing 2996 nonredundant total proteins. Subsequent multiple analyses (MapMan, CAZy, WallProtDB and expert curation) of this database were then used to identify a flax cell wall proteome consisting of 456 nonredundant proteins localized in the cell wall and/or associated with cell wall biosynthesis, remodeling and other cell wall related processes. Examination of the proteins present in different flax organs/tissues provided a detailed overview of cell wall metabolism and highlighted the importance of hemicellulose and pectin remodeling in stem tissues. Phylogenetic analyses of proteins in the cell wall proteome revealed an important paralogy in the class IIIA xyloglucan endo-transglycosylase/hydrolase (XTH) family associated with xyloglucan endo-hydrolase activity.Immunolocalisation, FT-IR microspectroscopy, and enzymatic fingerprinting indicated that flax fiber primary/S1 cell walls contained xyloglucans with typical substituted side chains as well as glucuronoxylans in much lower quantities. These results suggest a likely central role of xyloglucans and endotransglucosylase/hydrolase activity in flax fiber formation and cell wall remodeling processes. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. From Vegetable Waste to New Agents for Potential Health Applications: Antioxidant Properties and Effects of Extracts, Fractions and Pinocembrin from Glycyrrhiza glabra L. Aerial Parts on Viability of Five Human Cancer Cell Lines.

    PubMed

    Aiello, Francesca; Armentano, Biagio; Polerà, Nicoletta; Carullo, Gabriele; Loizzo, Monica Rosa; Bonesi, Marco; Cappello, Maria Stella; Capobianco, Loredana; Tundis, Rosa

    2017-09-13

    Glycyrrhiza glabra cultivation and harvesting produces substantial quantities of aerial parts as waste. With the aim to prospect an innovative valorization of these byproducts, the aerial parts were harvested in May and October and analyzed for their chemical profile, antioxidant properties, and effects on viability of five cancer cell lines. Pinocembrin was the main constituent. A significant protection of lipid peroxidation was observed with the May total extract (IC 50 of 4.2 ± 0.4 μg/mL at 30 min of incubation). The effects on viability of HeLa, MCF-7, MDA-MB-231, Caco-2, and PC3 human cancer cells were investigated. All samples shown a remarkable activity with IC 50 values below 25 μg/mL. Samples from plants harvested in May exhibited greater activity than those harvested in October. MCF-7 and HeLa were the most sensitive cells with IC 50 in the range 2.73-3.01 and 3.28-5.53 μg/mL, respectively. G. glabra aerial parts represent a good source of valuable products.

  14. Closed-form integrator for the quaternion (euler angle) kinematics equations

    NASA Technical Reports Server (NTRS)

    Whitmore, Stephen A. (Inventor)

    2000-01-01

    The invention is embodied in a method of integrating kinematics equations for updating a set of vehicle attitude angles of a vehicle using 3-dimensional angular velocities of the vehicle, which includes computing an integrating factor matrix from quantities corresponding to the 3-dimensional angular velocities, computing a total integrated angular rate from the quantities corresponding to a 3-dimensional angular velocities, computing a state transition matrix as a sum of (a) a first complementary function of the total integrated angular rate and (b) the integrating factor matrix multiplied by a second complementary function of the total integrated angular rate, and updating the set of vehicle attitude angles using the state transition matrix. Preferably, the method further includes computing a quanternion vector from the quantities corresponding to the 3-dimensional angular velocities, in which case the updating of the set of vehicle attitude angles using the state transition matrix is carried out by (a) updating the quanternion vector by multiplying the quanternion vector by the state transition matrix to produce an updated quanternion vector and (b) computing an updated set of vehicle attitude angles from the updated quanternion vector. The first and second trigonometric functions are complementary, such as a sine and a cosine. The quantities corresponding to the 3-dimensional angular velocities include respective averages of the 3-dimensional angular velocities over plural time frames. The updating of the quanternion vector preserves the norm of the vector, whereby the updated set of vehicle attitude angles are virtually error-free.

  15. A Reconstruction Approach to High-Order Schemes Including Discontinuous Galerkin for Diffusion

    NASA Technical Reports Server (NTRS)

    Huynh, H. T.

    2009-01-01

    We introduce a new approach to high-order accuracy for the numerical solution of diffusion problems by solving the equations in differential form using a reconstruction technique. The approach has the advantages of simplicity and economy. It results in several new high-order methods including a simplified version of discontinuous Galerkin (DG). It also leads to new definitions of common value and common gradient quantities at each interface shared by the two adjacent cells. In addition, the new approach clarifies the relations among the various choices of new and existing common quantities. Fourier stability and accuracy analyses are carried out for the resulting schemes. Extensions to the case of quadrilateral meshes are obtained via tensor products. For the two-point boundary value problem (steady state), it is shown that these schemes, which include most popular DG methods, yield exact common interface quantities as well as exact cell average solutions for nearly all cases.

  16. Cutting Edge: Protection by Antiviral Memory CD8 T Cells Requires Rapidly Produced Antigen in Large Amounts.

    PubMed

    Remakus, Sanda; Ma, Xueying; Tang, Lingjuan; Xu, Ren-Huan; Knudson, Cory; Melo-Silva, Carolina R; Rubio, Daniel; Kuo, Yin-Ming; Andrews, Andrew; Sigal, Luis J

    2018-05-15

    Numerous attempts to produce antiviral vaccines by harnessing memory CD8 T cells have failed. A barrier to progress is that we do not know what makes an Ag a viable target of protective CD8 T cell memory. We found that in mice susceptible to lethal mousepox (the mouse homolog of human smallpox), a dendritic cell vaccine that induced memory CD8 T cells fully protected mice when the infecting virus produced Ag in large quantities and with rapid kinetics. Protection did not occur when the Ag was produced in low amounts, even with rapid kinetics, and protection was only partial when the Ag was produced in large quantities but with slow kinetics. Hence, the amount and timing of Ag expression appear to be key determinants of memory CD8 T cell antiviral protective immunity. These findings may have important implications for vaccine design. Copyright © 2018 by The American Association of Immunologists, Inc.

  17. Effect of dexamethasone on expression of glucocorticoid receptor in human monocyte cell line THP-1.

    PubMed

    Li, Bo; Bai, Xiangjun; Wanh, Haiping

    2006-01-01

    The effect of dexamethasone with different concentrations and different stimulating periods on the expression of glucocorticoid receptors (GRalpha, GRbeta) protein was investigated in human monocyte cell line THP-1. The cultured human monocyte line THP-1 cells were stimulated by dexamethasone with different concentrations and different periods. The expression of GRalpha and GRbeta protein was detected by Western blotting. The results showed that the expression of GRalpha and GRbeta was detected in the THP-1 cells. The quantity of GRalpha expression was reduced by dexamethasone under the same concentration with the prolongation of the stimulating periods. The quantity of GRbeta expression was increased by dexamethasone treatment in a time- and dose-dependent manner. It was concluded that dexamethasone stimulation time-dependently reduced the GRalpha expression in THP-1 cells. Dexamethasone stimulation time- and dose-dependently increased the GRbeta expression in THP-1 cells. The expression of GRalpha and GRbeta was regulated by glucocorticoid.

  18. Elevated atmospheric CO2 affected photosynthetic products in wheat seedlings and biological activity in rhizosphere soil under cadmium stress.

    PubMed

    Jia, Xia; Liu, Tuo; Zhao, Yonghua; He, Yunhua; Yang, Mingyan

    2016-01-01

    The objective of this study was to investigate the effects of elevated CO2 (700 ± 23 μmol mol(-1)) on photosynthetic products in wheat seedlings and on organic compounds and biological activity in rhizosphere soil under cadmium (Cd) stress. Elevated CO2 was associated with decreased quantities of reducing sugars, starch, and soluble amino acids, and with increased quantities of soluble sugars, total sugars, and soluble proteins in wheat seedlings under Cd stress. The contents of total soluble sugars, total free amino acids, total soluble phenolic acids, and total organic acids in the rhizosphere soil under Cd stress were improved by elevated CO2. Compared to Cd stress alone, the activity of amylase, phenol oxidase, urease, L-asparaginase, β-glucosidase, neutral phosphatase, and fluorescein diacetate increased under elevated CO2 in combination with Cd stress; only cellulase activity decreased. Bacterial abundance in rhizosphere soil was stimulated by elevated CO2 at low Cd concentrations (1.31-5.31 mg Cd kg(-1) dry soil). Actinomycetes, total microbial abundance, and fungi decreased under the combined conditions at 5.31-10.31 mg Cd kg(-1) dry soil. In conclusion, increased production of soluble sugars, total sugars, and proteins in wheat seedlings under elevated CO2 + Cd stress led to greater quantities of organic compounds in the rhizosphere soil relative to seedlings grown under Cd stress only. Elevated CO2 concentrations could moderate the effects of heavy metal pollution on enzyme activity and microorganism abundance in rhizosphere soils, thus improving soil fertility and the microecological rhizosphere environment of wheat under Cd stress.

  19. Glide-plane symmetry and superconducting gap structure of iron-based superconductors

    DOE PAGES

    Wang, Yan; Berlijn, Tom; Hirschfeld, Peter J.; ...

    2015-03-10

    We consider the effect of glide-plane symmetry of the Fe-pnictogen/chalcogen layer in Fe-based superconductors on pairing in spin fluctuation models. Recent theories propose that so-called η-pairing states with nonzero total momentum can be realized and possess such exotic properties as odd parity spin singlet symmetry and time-reversal symmetry breaking. Here we show that when there is orbital weight at the Fermi level from orbitals with even and odd mirror reflection symmetry in z, η pairing is inevitable; however, we conclude from explicit calculation that the gap function appearing in observable quantities is identical to that found in earlier pseudocrystal momentummore » calculations with 1 Fe per unit cell.« less

  20. IMPACT OF FIVE TREATMENT FACTORS ON MUSSEL MORTALITY

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Daniel P. Molloy

    2003-12-08

    Under this USDOE-NETL contract, the bacterium Pseudomonas fluorescens is being developed as a biocontrol agent for zebra mussels. The specific purpose of the contract is to identify factors that affect mussel kill. Test results reported herein indicate that mussel kill should not be affected by: (1) air bubbles being carried by currents through power plant pipes; (2) pipe orientation (e.g., vertical or horizontal); (3) whether the bacterial cell concentration during a treatment is constant or slightly varying; (4) whether a treatment is between 3 hr and 12 hr in duration, given that the total quantity of bacteria being applied tomore » the pipe is a constant; and (5) whether the water temperature is between 13 C and 23 C.« less

  1. Development of an improved high efficiency thin silicon solar cell

    NASA Technical Reports Server (NTRS)

    Lindmayer, J.

    1978-01-01

    Efforts were concerned with optimizing techniques for thinning silicon slices in NaOH etches, initial investigations of surface texturing, variation of furnace treatments to improve cell efficiency, initial efforts on optimization of gridline and cell sizes and Pilot Line fabrication of quantities of 2 cm x 2 cm 50 micron thick cells.

  2. A Feasibility Study of Nonlinear Spectroscopic Measurement of Magnetic Nanoparticles Targeted to Cancer Cells.

    PubMed

    Ficko, Bradley W; NDong, Christian; Giacometti, Paolo; Griswold, Karl E; Diamond, Solomon G

    2017-05-01

    Magnetic nanoparticles (MNPs) are an emerging platform for targeted diagnostics in cancer. An important component needed for translation of MNPs is the detection and quantification of targeted MNPs bound to tumor cells. This study explores the feasibility of a multifrequency nonlinear magnetic spectroscopic method that uses excitation and pickup coils and is capable of discriminating between quantities of bound and unbound MNPs in 0.5 ml samples of KB and Igrov human cancer cell lines. The method is tested over a range of five concentrations of MNPs from 0 to 80 μg/ml and five concentrations of cells from 50 to 400 000 count per ml. A linear model applied to the magnetic spectroscopy data was able to simultaneously measure bound and unbound MNPs with agreement between the model-fit and lab assay measurements (p < 0.001). The detectable iron of the presented method to bound and unbound MNPs was < 2 μg in a 0.5 ml sample. The linear model parameters used to determine the quantities of bound and unbound nanoparticles in KB cells were also used to measure the bound and unbound MNP in the Igrov cell line and vice versa. Nonlinear spectroscopic measurement of MNPs may be a useful method for studying targeted MNPs in oncology. Determining the quantity of bound and unbound MNP in an unknown sample using a linear model represents an exciting opportunity to translate multifrequency nonlinear spectroscopy methods to in vivo applications where MNPs could be targeted to cancer cells.

  3. Triple point temperature of neon isotopes: Dependence on nitrogen impurity and sealed-cell model

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pavese, F.; Steur, P. P. M.; Giraudi, D.

    2013-09-11

    This paper illustrates a study conducted at INRIM, to further check how some quantities influence the value of the triple point temperature of the neon high-purity isotopes {sup 20}Ne and {sup 22}Ne. The influence of nitrogen as a chemical impurity in neon is critical with regard to the present best total uncertainty achieved in the measurement of these triple points, but only one determination is available in the literature. Checks are reported, performed on two different samples of {sup 22}Ne known to contain a N{sub 2} amount of 157⋅10{sup −6}, using two different models of sealed cells. The model ofmore » the cell can, in principle, have some effects on the shape of the melting plateau or on the triple point temperature observed for the sample sealed in it. This can be due to cell thermal parameters, or because the INRIM cell element mod. c contains many copper wires closely packed, which can, in principle, constrain the interface and induce a premelting-like effect. The reported results on a cell mod. Bter show no evident effect from the cell model and provide a value for the effect of N{sub 2} in Ne liquidus point of 8.6(1.9) μK ppm N{sub 2}{sup −1}, only slightly different from the literature datum.« less

  4. Multiple displacement amplification on single cell and possible PGD applications.

    PubMed

    Hellani, Ali; Coskun, Serdar; Benkhalifa, Moncef; Tbakhi, Abelghani; Sakati, Nadia; Al-Odaib, Ali; Ozand, Pinar

    2004-11-01

    Multiple displacement amplification (MDA) is a technique used in the amplification of very low amounts of DNA and reported to yield large quantities of high-quality DNA. We used MDA to amplify the whole genome directly from a single cell. The most common techniques used in PGD are PCR and fluorescent in-situ hybridization (FISH). There are many limitations to these techniques including, the number of chromosomes diagnosed for FISH or the quality of DNA issued from a single cell PCR. This report shows, for the first time, use of MDA for single cell whole genome amplification. A total of 16 short tandem repeats (STRs) were amplified successfully with a similar pattern to the genomic DNA. Furthermore, allelic drop out (ADO) derived from MDA was assessed in 40 single cells by analysing (i) heterozygosity for a known beta globin mutation (IVSI-5 C-G) and by studying (ii) the heterozygous loci present in the STRs. ADO turned out to be 10.25% for the beta globin gene sequencing and 5% for the fluorescent PCR analysis of STRs. Moreover, the amplification accuracy of MDA permitted the detection of trisomy 21 on a single cell using comparative genome hybridization-array. Altogether, these data suggest that MDA can be used for single cell molecular karyotyping and the diagnosis of any single gene disorder in PGD.

  5. Evaluation of p16 hypermethylation in oral submucous fibrosis: A quantitative and comparative analysis in buccal cells and saliva using real-time methylation-specific polymerase chain reaction.

    PubMed

    Kaliyaperumal, Subadra; Sankarapandian, Sathasivasubramanian

    2016-01-01

    The aim of this study was to quantitatively investigate the hypermethylation of p16 gene in buccal cells and saliva of oral submucous fibrosis (OSMF) patients using real-time quantitative methylation-specific polymerase chain reaction (PCR) and to compare the values of two methods. A total of 120 samples were taken from 60 subjects selected for this study, of which 30 were controls and 30 patients were clinically and histopathologically diagnosed with OSMF. In both groups, two sets of samples were collected, one directly from the buccal cells through cytobrush technique and the other through salivary rinse. We analyzed the samples for the presence of p16 hypermethylation using quantitative real-time PCR. In OSMF, the hypermethylation status of p16 in buccal cells was very high (93.3%) and in salivary samples, it was partially methylated (50%). However, no hypermethylation was found in controls suggesting that significant quantity of p16 hypermethylation was present in buccal cells and saliva in OSMF. This study indicates that buccal cell sampling may be a better method for evaluation than the salivary samples. It signifies that hypermethylation of p16 is an important factor to be considered in epigenetic alterations of normal cells to oral precancer, i.e. OSMF.

  6. Cell Expansion During Directed Differentiation of Stem Cells Toward the Hepatic Lineage.

    PubMed

    Raju, Ravali; Chau, David; Cho, Dong Seong; Park, Yonsil; Verfaillie, Catherine M; Hu, Wei-Shou

    2017-02-15

    The differentiation of human pluripotent stem cells toward the hepatocyte lineage can potentially provide an unlimited source of functional hepatocytes for transplantation and extracorporeal bioartificial liver applications. It is anticipated that the quantities of cells needed for these applications will be in the order of 10 9 -10 10 cells, because of the size of the liver. An ideal differentiation protocol would be to enable directed differentiation to the hepatocyte lineage with simultaneous cell expansion. We introduced a cell expansion stage after the commitment of human embryonic stem cells to the endodermal lineage, to allow for at least an eightfold increase in cell number, with continuation of cell maturation toward the hepatocyte lineage. The progressive changes in the transcriptome were measured by expression array, and the expression dynamics of certain lineage markers was measured by mass cytometry during the differentiation and expansion process. The findings revealed that while cells were expanding they were also capable of progressing in their differentiation toward the hepatocyte lineage. In addition, our transcriptome, protein and functional studies, including albumin secretion, drug-induced CYP450 expression and urea production, all indicated that the hepatocyte-like cells obtained with or without cell expansion are very similar. This method of simultaneous cell expansion and hepatocyte differentiation should facilitate obtaining large quantities of cells for liver cell applications.

  7. A Stable Human-Cell System Overexpressing Cystic Fibrosis Transmembrane Conductance Regulator Recombinant Protein at the Cell Surface

    PubMed Central

    Dai, Qun; Aleksandrov, Andrei A.; Bajrami, Bekim; Diego, Pamela Ann; Wu, Xing; Ray, Marjorie; Naren, Anjaparavanda P.; Riordan, John R.; Yao, Xudong; DeLucas, Lawrence J.; Urbatsch, Ina L.; Kappes, John C.

    2015-01-01

    Recent human clinical trials results demonstrated successful treatment for certain genetic forms of cystic fibrosis (CF). To extend treatment opportunities to those afflicted with other genetic forms of CF disease, structural and biophysical characterization of CF transmembrane conductance regulator (CFTR) is urgently needed. In this study, CFTR was modified with various tags, including a His10 purification tag, the SUMOstar (SUMO*) domain, an extracellular FLAG epitope, or an enhanced green fluorescent protein (EGFP), each alone or in various combinations. Expressed in HEK293 cells, recombinant CFTR proteins underwent complex glycosylation, compartmentalized with the plasma membrane, and exhibited regulated chloride-channel activity with only modest alterations in channel conductance and gating kinetics. Surface CFTR expression level was enhanced by the presence of SUMO* on the N-terminus. Quantitative mass-spectrometric analysis indicated approximately 10% of the total recombinant CFTR (SUMO*-CFTRFLAG-EGFP) localized to the plasma membrane. Trial purification using dodecylmaltoside for membrane protein extraction reproducibly recovered 178 ± 56 μg SUMO*-CFTRFLAG-EGFP per billion cells at 80% purity. Fluorescence size-exclusion chromatography indicated purified CFTR was monodisperse. These findings demonstrate a stable mammalian cell expression system capable of producing human CFTR of sufficient quality and quantity to augment futrure CF drug discovery efforts, including biophysical and structural studies. PMID:25577540

  8. Comparison of conceptually based and regression rainfall-runoff models, Denver Metropolitan area, Colorado, and potential applications in urban areas

    USGS Publications Warehouse

    Lindner-Lunsford, J. B.; Ellis, S.R.

    1987-01-01

    Multievent, conceptually based models and a single-event, multiple linear-regression model for estimating storm-runoff quantity and quality from urban areas were calibrated and verified for four small (57 to 167 acres) basins in the Denver metropolitan area, Colorado. The basins represented different land-use types - light commercial, single-family housing, and multi-family housing. Both types of models were calibrated using the same data set for each basin. A comparison was made between the storm-runoff volume, peak flow, and storm-runoff loads of seven water quality constituents simulated by each of the models by use of identical verification data sets. The models studied were the U.S. Geological Survey 's Distributed Routing Rainfall-Runoff Model-Version II (DR3M-II) (a runoff-quantity model designed for urban areas), and a multievent urban runoff quality model (DR3M-QUAL). Water quality constituents modeled were chemical oxygen demand, total suspended solids, total nitrogen, total phosphorus, total lead, total manganese, and total zinc. (USGS)

  9. Publication trends of research on diabetes mellitus and T cells (1997–2016): A 20-year bibliometric study

    PubMed Central

    He, Yifei; Chen, Rong; Zhou, Jingjing; Li, Ming

    2017-01-01

    Introduction Diabetes Mellitus (DM) is a huge burden for human health. Recent studies show the close relationship between DM and T cells. We investigated the trend in DM and T cells research. Methods Using the Web of Science database, we searched the publications on DM and T cells in 1997–2016, and studied the source data using bibliometric methodology. Excel 2016, GraphPad Prism 5, and VOSviewer software were used to analyze the publication trend in DM and T cells research. Results We found a total of 1077 publications with 38109 citations up to January 23, 2017. The highest contribution came from the United States, with 48.38% of the publications, 61.44% of the citations and the highest H-index (74). China had the 5th place for total publications, but ranked 11th both for citation frequency (604) and H-index (13). The inflection point of the global DM and T cells publications was in 2000. Journal of Immunology published the most related articles (164). Santamaria P. was the leading scholar in this field with the most publications (35). The keywords “regulatory T cell” and “autoimmune diabetes” were mentioned more than 300 times. Furthermore, type 2 (T2)DM, T cell immunoglobulin and mucin domain (TIM) and obesity are becoming popular research topics in this field. Conclusion The quantity of publications on DM and T cells grew rapidly around year 2000, but has relatively decreased recently. The United States had the leading position in global research. There was a discrepancy between productivity and quality of publications from China. Latest progress is most likely first published by the Journal of Immunology. Santamaria P., Roep B.O. and Peakman M. were the pioneer scholars in this field. Most researchers have focused on “regulatory T cell” and “autoimmune diabetes” research. In future, T2DM, TIM and obesity may be the popular areas. PMID:28926600

  10. Improving the assessment of prescribing: use of a 'substitution index'.

    PubMed

    Kunisawa, Susumu; Otsubo, Tetsuya; Lee, Jason; Imanaka, Yuichi

    2013-07-01

    To analyse the current and potential utilization of generic drugs in Japan, to examine the maximum possible cost savings from generic drug use and to develop a fairer measure to assess the level of generic drug substitution. We conducted a cross-sectional retrospective analysis of nine million dispensing records during January to March 2010 in Kyoto Prefecture. Maximum potential quantity-based shares were defined as the quantity of generic drugs used plus the quantity of branded drugs that could have been replaced by generic drugs divided by the quantity of all drugs dispensed. We developed a 'substitution index', defined as the proportion of generic drugs out of the total drugs substitutable with generic drugs (based on quantity rather than cost). Generic drugs had a quantity-based share of 17.9%, a cost-based share of 8.9% and a maximum potential quantity-based share of 50.1%, which is lower than the actual generic drug shares of some other countries. The maximum possible cost savings as a result of generic drug substitution was 16.5%. We also observed wide variations in maximum potential quantity-based shares between health care sectors and health care institutions. Simple comparisons based on quantity-based shares may misrepresent the actual generic drug use. A substitution index that takes into account the maximum potential quantity-based share of generic drugs as a fairer measure may promote more realistic goals and encourage generic drug usage.

  11. Laminin-111 improves skeletal muscle stem cell quantity and function following eccentric exercise.

    PubMed

    Zou, Kai; De Lisio, Michael; Huntsman, Heather D; Pincu, Yair; Mahmassani, Ziad; Miller, Matthew; Olatunbosun, Dami; Jensen, Tor; Boppart, Marni D

    2014-09-01

    Laminin-111 (α1, β1, γ1; LM-111) is an important component of the extracellular matrix that is required for formation of skeletal muscle during embryonic development. Recent studies suggest that LM-111 supplementation can enhance satellite cell proliferation and muscle function in mouse models of muscular dystrophy. The purpose of this study was to determine the extent to which LM-111 can alter satellite and nonsatellite stem cell quantity following eccentric exercise-induced damage in young adult, healthy mice. One week following injection of LM-111 or saline, mice either remained sedentary or were subjected to a single bout of downhill running (EX). While one muscle was preserved for evaluation of satellite cell number, the other muscle was processed for isolation of mesenchymal stem cells (MSCs; Sca-1+CD45-) via FACS at 24 hours postexercise. Satellite cell number was approximately twofold higher in LM-111/EX compared with all other groups (p<.05), and the number of satellite cells expressing the proliferation marker Ki67 was 50% to threefold higher in LM-111/EX compared with all other groups (p<.05). LM-111 also increased the quantity of embryonic myosin heavy chain-positive (eMHC+) fibers in young mice after eccentric exercise (p<.05). Although MSC percentage and number were not altered, MSC proinflammatory gene expression was decreased, and hepatocyte growth factor gene expression was increased in the presence of LM-111 (p<.05). Together, these data suggest that LM-111 supplementation provides a viable solution for increasing skeletal muscle stem cell number and/or function, ultimately allowing for improvements in the regenerative response to eccentric exercise. ©AlphaMed Press.

  12. Bone morphogenetic protein-mediated interaction of periosteum and diaphysis. Citric acid and other factors influencing the generation of parosteal bone.

    PubMed

    Kübler, N; Urist, M R

    1990-09-01

    In rabbits, after long-bone growth is complete and the cambium layer regresses, mesenchymal-type cells with embryonic potential (competence) for bone development persist in the adventitial layer of periosteum. These cells are not determined osteoprogenitor cells (stem cells) because bone tissue differentiation does not occur when adult periosteum is transplanted into a heterotopic site. In this respect, adventitial cells differ from bone marrow stroma cells. In a parosteal orthotopic site in the space between the adult periosteum and diaphysis, implants of bone morphogenetic protein (BMP) and associated noncollagenous proteins (BMP/NCP) induce adventitia and adjacent muscle connective-tissue-derived cells to switch from a fibrogenetic to a chondroosteoprogenetic pattern of bone development. The quantity of induced bone is proportional to the dose of BMP/NCP in the range from 10 to 50 mg; immature rabbits produced larger deposits than mature rabbits in response to BMP/NCP. Preoperative local intramuscular injections of citric, edetic, or hyaluronic acids in specified concentrations markedly enhanced subperiosteal BMP/NCP-induced bone formation. The quantity of bovine or human BMP/NCP-induced bone formation in rabbits is also increased by very low-dose immunosuppression but not by bone mineral, tricalcium phosphate ceramic, inorganic calcium salts, or various space-occupying, unspecific chemical irritants. Although composities of BMP/NCP and allogeneic rabbit tendon collagen increased the quantity of bone in a parosteal site, in a heterotopic site the composite failed to induce bone formation. In a parosteal site, the conditions permitting BMP/NCP-induced bone formation develop, and the end product of the morphogenetic response is a duplicate diaphysis. How BMP reactivates the morphogenetic process in postfetal mesenchymal-type adventitial cells persisting in adult periosteum (including adjacent muscle attachments) is not known.

  13. Association between Apoptotis and CD4+/CD8+ T-Lymphocyte Ratio in Aseptic Loosening after Total Hip Replacement

    PubMed Central

    Landgraeber, Stefan; von Knoch, Marius; Löer, Franz; Brankamp, Jochen; Tsokos, Michael; Grabellus, Florian; Schmid, Kurt Werner; Totsch, Martin

    2009-01-01

    Particle-induced osteolysis is a major cause of aseptic loosening after total joint replacement. While the osteolytic cascade initiated by cytokine release from macrophages has been studied extensively, the involvement of T-lymphocytes in this context is controversial and has been addressed by only a few authors. In a former study we detected that the quantity of T-lymphocytes may be influenced by apoptosis in patients with aseptic loosening. In this study we intended to find out more details about the apoptosis-induced shifting of the T-cell number. We focused our interest on the CD4+ and CD8+ T-cells and their relative ratio. Caspase-3 cleaved was evaluated immunohistochemically to detect apoptotic T-cells in capsules and interface membranes from patients with aseptic hip implant loosening and a varying degree of caspase-3 cleaved expression in CD4+ and CD8+ T-lymphocytes was detected. Moreover, a relationship between the intensity of the apoptotic reactions and the radiological extent of osteolysis was observed. The number of CD4+ cells was decreased in the presence of strong apoptotic reactions, respectively extensive osteolysis, while CD8+ cells were affected to a much lower degree. Thus, the CD4+/CD8+ ratio changed from 1.0 in cases with only small areas of periprosthetic osteolysis and minimally intense apoptosis to 0.33 in cases with large areas of osteolysis. This may suggest a causal relationship between the apoptosis-induced shift in the CD4+/CD8+ ratio and the osteolysis respectively aseptic loosening. It is possible that these findings may lead to a new understanding of particle-induced osteolysis. PMID:19214244

  14. A study of waste liquid crystal display generation in mainland China.

    PubMed

    Liu, Zhifeng; Xu, Zeying; Huang, Haihong; Li, Bingbing

    2016-01-01

    The generation of liquid crystal display waste is becoming a serious social problem. Predicting liquid crystal display waste status is the foundation for establishing a recycling network; however, the difficulty in predicting liquid crystal display waste quantity lies in data mining. In order to determine the quantity and the distribution of liquid crystal display waste in China, the four top-selling liquid crystal display products (liquid crystal display TVs, desktop PCs, notebook PCs, and mobile phones) were selected as study objects. Then, the extended logistic model and market supply A method was used to predict the quantity of liquid crystal display waste products. Moreover, the distribution of liquid crystal display waste products in different regions was evaluated by examining the consumption levels of household equipment. The results revealed that the quantity of waste liquid crystal displays would increase rapidly in the next decade. In particular, the predicted quantity of waste liquid crystal displays would rise to approximately 4.262 × 10(9) pieces in 2020, and the total display area (i.e. the surface area of liquid crystal display panels) of waste liquid crystal displays would reach 5.539 × 10(7) m(2). The prediction on the display area of waste liquid crystal display TVs showed that it would account for 71.5% of the total display area by 2020. Meanwhile, the quantity of waste mobile phones would significantly grow, increasing 5.8 times from 2012 to 2020. In terms of distribution, Guangdong is the top waste liquid crystal display-generating province in China, followed by Jiangsu, Shandong, Henan, Zhejiang, and Sichuan. Considering its regional characteristics, Guangdong has been proposed to be the most important location of the recycling network. © The Author(s) 2015.

  15. Human Milk MicroRNA and Total RNA Differ Depending on Milk Fractionation.

    PubMed

    Alsaweed, Mohammed; Hepworth, Anna R; Lefèvre, Christophe; Hartmann, Peter E; Geddes, Donna T; Hassiotou, Foteini

    2015-10-01

    MicroRNA have been recently discovered in human milk signifying potentially important functions for both the lactating breast and the infant. Whilst human milk microRNA have started to be explored, little data exist on the evaluation of sample processing, and analysis to ensure that a full spectrum of microRNA can be obtained. Human milk comprises three main fractions: cells, skim milk, and lipids. Typically, the skim milk fraction has been measured in isolation despite evidence that the lipid fraction may contain more microRNA. This study aimed to standardize isolation of microRNA and total RNA from all three fractions of human milk to determine the most appropriate sampling and analysis procedure for future studies. Three different methods from eight commercially available kits were tested for their efficacy in extracting total RNA and microRNA from the lipid, skim, and cell fractions of human milk. Each fraction yielded different concentrations of RNA and microRNA, with the highest quantities found in the cell and lipid fractions, and the lowest in skim milk. The column-based phenol-free method was the most efficient extraction method for all three milk fractions. Two microRNAs were expressed and validated in the three milk fractions by qPCR using the three recommended extraction kits for each fraction. High expression levels were identified in the skim and lipid milk factions for these microRNAs. These results suggest that careful consideration of both the human milk sample preparation and extraction protocols should be made prior to embarking upon research in this area. © 2015 The Authors. Journal of Cellular Biochemistry Published by Wiley Periodicals, Inc.

  16. Optimal ordering quantities for substitutable deteriorating items under joint replenishment with cost of substitution

    NASA Astrophysics Data System (ADS)

    Mishra, Vinod Kumar

    2017-09-01

    In this paper we develop an inventory model, to determine the optimal ordering quantities, for a set of two substitutable deteriorating items. In this inventory model the inventory level of both items depleted due to demands and deterioration and when an item is out of stock, its demands are partially fulfilled by the other item and all unsatisfied demand is lost. Each substituted item incurs a cost of substitution and the demands and deterioration is considered to be deterministic and constant. Items are order jointly in each ordering cycle, to take the advantages of joint replenishment. The problem is formulated and a solution procedure is developed to determine the optimal ordering quantities that minimize the total inventory cost. We provide an extensive numerical and sensitivity analysis to illustrate the effect of different parameter on the model. The key observation on the basis of numerical analysis, there is substantial improvement in the optimal total cost of the inventory model with substitution over without substitution.

  17. An economic order quantity model with shortage and inflation

    NASA Astrophysics Data System (ADS)

    Wulan, Elis Ratna; Nurjaman, Wildan

    2015-09-01

    The effect of inflation has become a persistent characteristic and more significant problem of many developing economies especially in the third world countries. While making effort to achieve optimal quantity of product to be produced or purchased using the simplest and on the shelf classical EOQ model, the non-inclusion of conflicting economic realities as shortage and inflation has rendered its result quite uneconomical and hence the purpose for this study. Mathematical expression was developed for each of the cost components the sum of which become the total inventory model over the period (0,L) ((TIC(0,L)). L is planning horizon and TIC(0,L) is total inventory cost over a period of (0,L). Significant savings with increase in quantity was achieved based on deference in the varying price regime. With the assumptions considered and subject to the availability of reliable inventory cost element, the developed model is found to produce a feasible, and economic inventory stock-level with the numerical example of a material supply of a manufacturing company.

  18. Extraction of extracellular polymeric substances from aerobic granule with compact interior structure.

    PubMed

    Adav, Sunil S; Lee, Duu-Jong

    2008-06-15

    Extracellular polymeric substances (EPS) were extracted from aerobic granules of compact interior structure using seven extraction methods. Ultrasound followed by the chemical reagents formamide and NaOH outperformed other methods in extracting EPS from aerobic granules of compact interior. The collected EPS revealed no contamination by intracellular substances and consisted mainly of proteins, polysaccharides, humic substances and lipids. The quantity of extracted proteins exhibited a weak correlation with quantity of extracted carbohydrates but no correlation with quantity of extracted humic substances. The total polysaccharides/total proteins (PN/PS) ratios for sludge flocs were approximately 0.9 regardless of extraction method. Protein content was significantly enriched in the granules, producing a PN/PS ratio of 3.4-6.2. This experimental result correlated with observations using excitation-emission matrix (EEM) and confocal laser scanning microscope technique. However, detailed study disproved the use of EEM results as a quantitative index of extracted EPS from sludge flocs or from granules.

  19. Methodology to Estimate the Quantity, Composition, and ...

    EPA Pesticide Factsheets

    This report, Methodology to Estimate the Quantity, Composition and Management of Construction and Demolition Debris in the US, was developed to expand access to data on CDD in the US and to support research on CDD and sustainable materials management. Since past US EPA CDD estimates have been limited to building-related CDD, a goal in the development of this methodology was to use data originating from CDD facilities and contractors to better capture the current picture of total CDD management, including materials from roads, bridges and infrastructure. This report, Methodology to Estimate the Quantity, Composition and Management of Construction and Demolition Debris in the US, was developed to expand access to data on CDD in the US and to support research on CDD and sustainable materials management. Since past US EPA CDD estimates have been limited to building-related CDD, a goal in the development of this methodology was to use data originating from CDD facilities and contractors to better capture the current picture of total CDD management, including materials from roads, bridges and infrastructure.

  20. Proteomic analysis of Chlorella vulgaris: Potential targets for enhanced lipid accumulation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Guarnieri, Michael T.; Nag, Ambarish; Yang, Shihui

    2013-11-01

    Oleaginous microalgae are capable of producing large quantities of fatty acids and triacylglycerides. As such, they are promising feedstocks for the production of biofuels and bioproducts. Genetic strain-engineering strategies offer a means to accelerate the commercialization of algal biofuels by improving the rate and total accumulation of microalgal lipids. However, the industrial potential of these organisms remains to be met, largely due to the incomplete knowledgebase surrounding the mechanisms governing the induction of algal lipid biosynthesis. Such strategies require further elucidation of genes and gene products controlling algal lipid accumulation. In this study, we have set out to examine thesemore » mechanisms and identify novel strain-engineering targets in the oleaginous microalga, Chlorella vulgaris. Comparative shotgun proteomic analyses have identified a number of novel targets, including previously unidentified transcription factors and proteins involved in cell signaling and cell cycle regulation. These results lay the foundation for strain-improvement strategies and demonstrate the power of translational proteomic analysis.« less

  1. Synthesis of milligram quantities of proteins using a reconstituted in vitro protein synthesis system.

    PubMed

    Kazuta, Yasuaki; Matsuura, Tomoaki; Ichihashi, Norikazu; Yomo, Tetsuya

    2014-11-01

    In this study, the amount of protein synthesized using an in vitro protein synthesis system composed of only highly purified components (the PURE system) was optimized. By varying the concentrations of each system component, we determined the component concentrations that result in the synthesis of 0.38 mg/mL green fluorescent protein (GFP) in batch mode and 3.8 mg/mL GFP in dialysis mode. In dialysis mode, protein concentrations of 4.3 and 4.4 mg/mL were synthesized for dihydrofolate reductase and β-galactosidase, respectively. Using the optimized system, the synthesized protein represented 30% (w/w) of the total protein, which is comparable to the level of overexpressed protein in Escherichia coli cells. This optimized reconstituted in vitro protein synthesis system may potentially be useful for various applications, including in vitro directed evolution of proteins, artificial cell assembly, and protein structural studies. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  2. k-Space Image Correlation Spectroscopy: A Method for Accurate Transport Measurements Independent of Fluorophore Photophysics

    PubMed Central

    Kolin, David L.; Ronis, David; Wiseman, Paul W.

    2006-01-01

    We present the theory and application of reciprocal space image correlation spectroscopy (kICS). This technique measures the number density, diffusion coefficient, and velocity of fluorescently labeled macromolecules in a cell membrane imaged on a confocal, two-photon, or total internal reflection fluorescence microscope. In contrast to r-space correlation techniques, we show kICS can recover accurate dynamics even in the presence of complex fluorophore photobleaching and/or “blinking”. Furthermore, these quantities can be calculated without nonlinear curve fitting, or any knowledge of the beam radius of the exciting laser. The number densities calculated by kICS are less sensitive to spatial inhomogeneity of the fluorophore distribution than densities measured using image correlation spectroscopy. We use simulations as a proof-of-principle to show that number densities and transport coefficients can be extracted using this technique. We present calibration measurements with fluorescent microspheres imaged on a confocal microscope, which recover Stokes-Einstein diffusion coefficients, and flow velocities that agree with single particle tracking measurements. We also show the application of kICS to measurements of the transport dynamics of α5-integrin/enhanced green fluorescent protein constructs in a transfected CHO cell imaged on a total internal reflection fluorescence microscope using charge-coupled device area detection. PMID:16861272

  3. Impact of tissue type and content of neoplastic cells of samples on the quality of epidermal growth factor receptor mutation analysis among patients with lung adenocarcinoma

    PubMed Central

    PALIOGIANNIS, PANAGIOTIS; ATTENE, FEDERICO; COSSU, ANTONIO; DEFRAIA, EFISIO; PORCU, GIUSEPPE; CARTA, ANNAMARIA; SOTGIU, MARIA IGNAZIA; PAZZOLA, ANTONIO; CORDERO, LORENZO; CAPELLI, FRANCESCA; FADDA, GIOVANNI MARIA; ORTU, SALVATORE; SOTGIU, GIOVANNI; PALOMBA, GRAZIA; SINI, MARIA CRISTINA; PALMIERI, GIUSEPPE; COLOMBINO, MARIA

    2015-01-01

    Assessment of the epidermal growth factor receptor (EGFR) mutational status has become crucial in recent years in the molecular classification of patients with lung cancer. The impact of the type and quantity of malignant cells of the neoplastic specimen on the quality of mutation analysis remains to be elucidated, and only empirical and sporadic data are available. The aim of the present study was to investigate the impact of tissue type and content of neoplastic cells in the specimen on the quality of EGFR mutation analysis among patients with lung adenocarcinoma. A total of 515 patients with histologically-confirmed disease were included in the present study. Formalin-fixed paraffin embedded tissue samples were used for the mutation analysis and the content of the neoplastic cells was evaluated using light microscopy. Genomic DNA was isolated using a standard protocol. The coding sequences and splice junctions of exons 18, 19 and 21 in the EGFR gene were then screened for mutations by direct automated sequencing. The mean age of the patients examined was 64.9 years and 357 (69.3%) were male. A total of 429 tissue samples (83.3%) were obtained by biopsy and the remaining samples were obtained by surgery. A total of 456 samples (88.5%) were observed from primary lung adenocarcinomas, while 59 (11.5%) were from metastatic lesions. EGFR mutations occurred in 59 cases (11.5%); exon 18 mutations were detected in one case (1.7%), whereas exon 19 and 21 mutations were detected in 30 (51%) and 28 (47.3%) cases, respectively. EGFR mutations were more frequent in females and patients that had never smoked. The distribution of the mutations among primary and metastatic tissues exhibited no significant differences in the proportions of EGFR mutations detected. However, a statistically significant difference in the number of mutations detected was found between samples with at least 50% of neoplastic cells (450 cases-57 mutations; 12.7%) and those with <50% of neoplastic cells (65 cases-2 mutations; 3.1%). PMID:25683726

  4. Climate Influences the Content and Chemical Composition of Foliar Tannins in Green and Senesced Tissues of Quercus rubra

    PubMed Central

    Top, Sara M.; Preston, Caroline M.; Dukes, Jeffrey S.; Tharayil, Nishanth

    2017-01-01

    Environmental stresses not only influence production of plant metabolites but could also modify their resorption during leaf senescence. The production-resorption dynamics of polyphenolic tannins, a class of defense compound whose ecological role extends beyond tissue senescence, could amplify the influence of climate on ecosystem processes. We studied the quantity, chemical composition, and tissue-association of tannins in green and freshly-senesced leaves of Quercus rubra exposed to different temperature (Warming and No Warming) and precipitation treatments (Dry, Ambient, Wet) at the Boston-Area Climate Experiment (BACE) in Massachusetts, USA. Climate influenced not only the quantity of tannins, but also their molecular composition and cell-wall associations. Irrespective of climatic treatments, tannin composition in Q. rubra was dominated by condensed tannins (CTs, proanthocyanidins). When exposed to Dry and Ambient*Warm conditions, Q. rubra produced higher quantities of tannins that were less polymerized. In contrast, under favorable conditions (Wet), tannins were produced in lower quantities, but the CTs were more polymerized. Further, even as the overall tissue tannin content declined, the content of hydrolysable tannins (HTs) increased under Wet treatments. The molecular composition of tannins influenced their content in senesced litter. Compared to the green leaves, the content of HTs decreased in senesced leaves across treatments, whereas the CT content was similar between green and senesced leaves in Wet treatments that produced more polymerized tannins. The content of total tannins in senesced leaves was higher in Warming treatments under both dry and ambient precipitation treatments. Our results suggest that, though climate directly influenced the production of tannins in green tissues (and similar patterns were observed in the senesced tissue), the influence of climate on tannin content of senesced tissue was partly mediated by the effect on the chemical composition of tannins. These different climatic impacts on leaves over the course of a growing season may alter forest dynamics, not only in decomposition and nutrient cycling dynamics, but also in herbivory dynamics. PMID:28559896

  5. Mutant quantity and quality in mammalian cells (A{sub L}) exposed to cesium-137 gamma radiation: Effect of caffeine

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    McGuinness, S.M.; Shibuya, M.L.; Ueno, A.M.

    1995-06-01

    We examined the effect of caffeine (1,3,7-trimethylxanthine) on the quantity and quality of mutations in cultured mammalian A{sub L} human-hamster hybrid cells exposed to {sup 137}Cs {gamma} radiation. At a dose (1.5 mg/ml for 16 h) that reduced the plating efficiency (PE) by 20%, caffeine was not itself a significant mutagen, but it increased by approximately twofold the slope of the dose-response curve for induction of S1{sup {minus}} mutants by {sup 137}Cs {gamma} radiation. Molecular analysis of 235 S1{sup {minus}} mutants using a series of DNA probes mapped to the human chromosome 11 in the A{sub L} hybrid cells revealedmore » that 73 to 85% of the mutations in unexposed cells and in cells treated with caffeine alone, {sup 137}Cs {gamma} rays alone or {sup 137}Cs {gamma} rays plus caffeine were large deletions involving millions of base pairs of DNA. Most of these deletions were contiguous with the region of the MIC1 gene at 11p13 that encodes the S1 cell surface antigen. In other mutants that had suffered multiple marker loss, the deletions were intermittent along chromosome 11. These {open_quotes}complex{close_quotes} mutations were rare for {sup 137}Cs {gamma} irradiation (1/63 = 1.5%) but relatively prevalent (23-50%) for other exposure conditions. Thus caffeine appears to alter both the quantity and quality of mutations induced by {sup 137}Cs {gamma} irradiation. 62 refs., 3 figs., 3 tabs.« less

  6. Conserved Quantities in General Relativity: From the Quasi-Local Level to Spatial Infinity

    NASA Astrophysics Data System (ADS)

    Chen, Po-Ning; Wang, Mu-Tao; Yau, Shing-Tung

    2015-08-01

    We define quasi-local conserved quantities in general relativity by using the optimal isometric embedding in Wang and Yau (Commun Math Phys 288(3):919-942, 2009) to transplant Killing fields in the Minkowski spacetime back to the 2-surface of interest in a physical spacetime. To each optimal isometric embedding, a dual element of the Lie algebra of the Lorentz group is assigned. Quasi-local angular momentum and quasi-local center of mass correspond to pairing this element with rotation Killing fields and boost Killing fields, respectively. They obey classical transformation laws under the action of the Poincaré group. We further justify these definitions by considering their limits as the total angular momentum and the total center of mass of an isolated system. These expressions were derived from the Hamilton-Jacobi analysis of the gravitational action and thus satisfy conservation laws. As a result, we obtained an invariant total angular momentum theorem in the Kerr spacetime. For a vacuum asymptotically flat initial data set of order 1, it is shown that the limits are always finite without any extra assumptions. We also study these total conserved quantities on a family of asymptotically flat initial data sets evolving by the vacuum Einstein evolution equation. It is shown that the total angular momentum is conserved under the evolution. For the total center of mass, the classical dynamical formula relating the center of mass, energy, and linear momentum is recovered, in the nonlinear context of initial data sets evolving by the vacuum Einstein evolution equation. The definition of quasi-local angular momentum provides an answer to the second problem in classical general relativity on Penrose's list (Proc R Soc Lond Ser A 381(1780):53-63, 1982).

  7. Determination of the Composition and Quantity of Phthalate Ester Additives in PVC Children's Toys. Greenpeace Research Laboratories Technical Note 06/97.

    ERIC Educational Resources Information Center

    Stringer, Ruth; Labounskaia, Irina; Santillo, David; Johnston, Paul; Siddorn, John; Stephenson, Angela

    Polyvinyl chloride (vinyl or PVC) is widely used in toys and other children's products. This study, conducted by Greenpeace, examined the composition and quantity of phthalate ester additives in children's PVC toys, used to give the toys added flexibility. Drawn from 17 countries, a total of 71 toys designed to be chewed by babies and young…

  8. Blood cell mRNAs and microRNAs: optimized protocols for extraction and preservation.

    PubMed

    Eikmans, Michael; Rekers, Niels V; Anholts, Jacqueline D H; Heidt, Sebastiaan; Claas, Frans H J

    2013-03-14

    Assessing messenger RNA (mRNA) and microRNA levels in peripheral blood cells may complement conventional parameters in clinical practice. Working with small, precious samples requires optimal RNA yields and minimal RNA degradation. Several procedures for RNA extraction and complementary DNA (cDNA) synthesis were compared for their efficiency. The effect on RNA quality of freeze-thawing peripheral blood cells and storage in preserving reagents was investigated. In terms of RNA yield and convenience, quality quantitative polymerase chain reaction signals per nanogram of total RNA and using NucleoSpin and mirVana columns is preferable. The SuperScript III protocol results in the highest cDNA yields. During conventional procedures of storing peripheral blood cells at -180°C and thawing them thereafter, RNA integrity is maintained. TRIzol preserves RNA in cells stored at -20°C. Detection of mRNA levels significantly decreases in degraded RNA samples, whereas microRNA molecules remain relatively stable. When standardized to reference targets, mRNA transcripts and microRNAs can be reliably quantified in moderately degraded (quality index 4-7) and severely degraded (quality index <4) RNA samples, respectively. We describe a strategy for obtaining high-quality and quantity RNA from fresh and stored cells from blood. The results serve as a guideline for sensitive mRNA and microRNA expression assessment in clinical material.

  9. Dexamethasone alone and in combination with desipramine, phenytoin, valproic acid or levetiracetam interferes with 5-ALA-mediated PpIX production and cellular retention in glioblastoma cells.

    PubMed

    Lawrence, Johnathan E; Steele, Christopher J; Rovin, Richard A; Belton, Robert J; Winn, Robert J

    2016-03-01

    Extent of resection of glioblastoma (GBM) correlates with overall survival. Fluorescence-guided resection (FGR) using 5-aminolevulinic acid (5-ALA) can improve the extent of resection. Unfortunately not all patients given 5-ALA accumulate sufficient quantities of protoporphyrin IX (PpIX) for successful FGR. In this study, we investigated the effects of dexamethasone, desipramine, phenytoin, valproic acid, and levetiracetam on the production and accumulation of PpIX in U87MG cells. All of these drugs, except levetiracetam, reduce the total amount of PpIX produced by GBM cells (p < 0.05). When dexamethasone is mixed with another drug (desipramine, phenytoin, valproic acid or levetiracetam) the amount of PpIX produced is further decreased (p < 0.01). However, when cells are analyzed for PpIX cellular retention, dexamethasone accumulated significantly more PpIX than the vehicle control (p < 0.05). Cellular retention of PpIX was not different from controls in cells treated with dexamethasone plus desipramine, valproic acid or levetiracetam, but was significantly less for dexamethasone plus phenytoin (p < 0.01). These data suggest that medications given before and during surgery may interfere with PpIX accumulation in malignant cells. At this time, levetiracetam appears to be the best medication in its class (anticonvulsants) for patients undergoing 5-ALA-mediated FGR.

  10. Development of Probiotic Formulation for the Treatment of Iron Deficiency Anemia.

    PubMed

    Korčok, Davor Jovan; Tršić-Milanović, Nada Aleksandar; Ivanović, Nevena Djuro; Đorđević, Brižita Ivan

    2018-04-01

    Probiotics are increasingly more present both as functional foods, and in pharmaceutical preparations with multiple levels of action that contribute to human health. Probiotics realize their positive effects with a proper dose, and by maintaining a declared number of probiotics cells by the expiration date. Important precondition for developing a probiotic product is the right choice of clinically proven probiotic strain, the choice of other active components, as well as, the optimization of the quantity of active component of probiotic per product dose. This scientific paper describes the optimization of the number of probiotics cells in the formulation of dietary supplement that contains probiotic culture Lactobacillus plantarum 299v, iron and vitamin C. Variations of the quantity of active component were analyzed in development batches of the encapsulated probiotic product categorized as dietary supplement with the following ingredients: probiotic culture, sucrosomal form of iron and vitamin C. Optimal quantity of active component L. plantarum of 50 mg, was selected. The purpose of this scientific paper is to select the optimal formulation of probiotic culture in a dietary supplement that contains iron and vitamin C, and to also determine its expiration date by the analysis of the number of viable probiotic cells.

  11. Daily changes of structure, function and rhodopsin content in the compound eye of the crab Hemigrapsus sanguineus.

    PubMed

    Arikawa, K; Kawamata, K; Suzuki, T; Eguchi, E

    1987-08-01

    The compound eye of the crab hemigrapsus sanguineus undergoes daily changes in morphology as determined by light and electron microscopy, both in the quantity of chromophore substances studied by HPLC and in visual sensitivity as shown by electrophysiological techniques. 1. At a temperature of 20 degrees C, the rhabdom occupation ratio (ROR) of an ommatidial retinula was 11.6% (maximum) at midnight, 8.0 times larger than the minimum value at midday (1.4%). 2. Observations by freeze-fracture revealed that the densities of intra-membranous particles (9-11 nm in diameter) of rhabdomeric membrane were ca. 2000/microns 2 and ca. 3000/microns 2 for night and daytime compound eyes, respectively. 3. Screening pigment granules migrated longitudinally and aggregated at night, but dispersed during the day. Reflecting pigment granules migrate transversally in the proximal half of the reticula layer i.e. cytoplasmic extensions containing reflecting pigment granules squeeze between neighbouring retinula cells causing optical isolation (Fig. 4). Thus the screening pigment granules within the retinula cells show longitudinal migration and radial movement so that the daytime rhabdoms are closely surrounded by the pigment granules. 4. At 20 degrees C, the total amount of chromophore of the visual pigment (11-cis and all-trans-retinal) was 1.4 times larger at night than during the day i.e. 46.6 pmol/eye at midnight and 33.2 pmol/eye at midday. Calculations of the total surface area of rhabdomeric membrane, total number of intra-membranous particles in rhabdomeric membrane and the total number of chromophore molecules in a compound eye, indicate that a considerable amount of chromophore-protein complex exists outside the rhabdom during the day. 5. The change in rhabdom size and quantity of chromophore were highly dependent on temperature. At 10 degrees C both rhabdom size and amount of chromophore stayed close to daytime levels throughout the 24 hours. 6. The intracellularly determined relative sensitivity of the dark adapted night eye to a point source of light was about twice as high as the dark-adapted day eye. Most of the increase in the sensitivity is attributed primarily to the effect of reflecting pigment migration around the basement membrane and, secondarily, to the changes in the amount and properties of the photoreceptive membrane. The results form the basis of a detailed discussion as to how an apposition eye can function possibly as a night-eye.

  12. Protein Folding and the Challenges of Maintaining Endoplasmic Reticulum Proteostasis in Idiopathic Pulmonary Fibrosis.

    PubMed

    Romero, Freddy; Summer, Ross

    2017-11-01

    Alveolar epithelial type II (AEII) cells are "professional" secretory cells that synthesize and secrete massive quantities of proteins to produce pulmonary surfactant and maintain airway immune defenses. To facilitate this high level of protein synthesis, AEII cells are equipped with an elaborate endoplasmic reticulum (ER) structure and possess an abundance of the machinery needed to fold, assemble, and secrete proteins. However, conditions that suddenly increase the quantity of new proteins entering the ER or that impede the capacity of the ER to fold proteins can cause misfolded or unfolded proteins to accumulate in the ER lumen, also called ER stress. To minimize this stress, AEII cells adapt by (1) reducing the quantity of proteins entering the ER, (2) increasing the amount of protein-folding machinery, and (3) removing misfolded proteins when they accumulate. Although these adaptive responses, aptly named the unfolded protein response, are usually effective in reducing ER stress, chronic aggregation of misfolded proteins is recognized as a hallmark feature of AEII cells in patients with idiopathic pulmonary fibrosis (IPF). Although mutations in surfactant proteins are linked to the development of ER stress in some rare IPF cases, the mechanisms causing protein misfolding in most cases are unknown. In this article, we review the mechanisms regulating ER proteostasis and highlight specific aspects of protein folding and the unfolded protein response that are most vulnerable to failure. Then, we postulate mechanisms other than genetic mutations that might contribute to protein aggregation in the alveolar epithelium of IPF lung.

  13. 40 CFR 98.434 - Monitoring and QA/QC requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Contained in Pre-Charged Equipment or Closed-Cell Foams § 98.434 Monitoring and QA/QC requirements. (a) For... equipment or closed-cell foam in the correct quantities (metric tons) and units (kg per piece of equipment...

  14. 40 CFR 98.434 - Monitoring and QA/QC requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Contained in Pre-Charged Equipment or Closed-Cell Foams § 98.434 Monitoring and QA/QC requirements. (a) For... equipment or closed-cell foam in the correct quantities (metric tons) and units (kg per piece of equipment...

  15. 40 CFR 98.434 - Monitoring and QA/QC requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Contained in Pre-Charged Equipment or Closed-Cell Foams § 98.434 Monitoring and QA/QC requirements. (a) For... equipment or closed-cell foam in the correct quantities and units. [74 FR 56374, Oct. 30, 2009, as amended...

  16. Liquid Based Preparation (LBP) cytology versus Conventional Cytology (CS) in FNA samples from breast, thyroid, salivary glands and soft tissues. Our experience in Crete (Greece).

    PubMed

    Mygdakos, N; Nikolaidou, Sylvia; Tzilivaki, Anna; Tamiolakis, D

    2009-01-01

    The improvement in quality of cytological preparations with the use of LBP methodology has been well-documented, but the cytological artifacts resulting from this technique have not been adequately described. This study describes and illustrates the cytological artifacts introduced by LBP technique when used on fine-needle aspirates (FNAs), and evaluates these artifacts as potential diagnostic pitfalls. We reviewed a total of 96 FNAs simultaneously processed by both conventional smears and LBP. FNAs were obtained from the following sites: lymph node (38), breast (28), soft-tissue sites (nine), salivary glands (six), and thyroid gland (15). The LBP smears were consistently devoid of obscuring elements, and the cells were adequately preserved and evenly dispersed. However, we noted some cytomorphological alterations that should be recognized to avoid erroneous diagnoses. The size of cell clusters was decreased, large branching sheets were fragmented, and there were more single cells, resulting in apparent discohesion. Small cells such as lymphocytes tended to aggregate. All cells were generally smaller and occasionally spindled, the chromatin detail was attenuated, and nucleoli were more prominent. Intranuclear inclusions were difficult to visualize. Background matrix was often altered in both quantity and quality. Extracellular particles, small mononuclear cells, red blood cells, and myoepithelial cells were markedly decreased in number. Cytopathologists should be careful in interpreting FNAs prepared using LBP technique if that is the only methodology employed. Familiarity with artifacts is essential to avoid misdiagnoses.

  17. Unexpected seasonality in quantity and composition of Amazon rainforest air reactivity

    PubMed Central

    Nölscher, A. C.; Yañez-Serrano, A. M.; Wolff, S.; de Araujo, A. Carioca; Lavrič, J. V.; Kesselmeier, J.; Williams, J.

    2016-01-01

    The hydroxyl radical (OH) removes most atmospheric pollutants from air. The loss frequency of OH radicals due to the combined effect of all gas-phase OH reactive species is a measureable quantity termed total OH reactivity. Here we present total OH reactivity observations in pristine Amazon rainforest air, as a function of season, time-of-day and height (0–80 m). Total OH reactivity is low during wet (10 s−1) and high during dry season (62 s−1). Comparison to individually measured trace gases reveals strong variation in unaccounted for OH reactivity, from 5 to 15% missing in wet-season afternoons to mostly unknown (average 79%) during dry season. During dry-season afternoons isoprene, considered the dominant reagent with OH in rainforests, only accounts for ∼20% of the total OH reactivity. Vertical profiles of OH reactivity are shaped by biogenic emissions, photochemistry and turbulent mixing. The rainforest floor was identified as a significant but poorly characterized source of OH reactivity. PMID:26797390

  18. Unexpected seasonality in quantity and composition of Amazon rainforest air reactivity.

    PubMed

    Nölscher, A C; Yañez-Serrano, A M; Wolff, S; de Araujo, A Carioca; Lavrič, J V; Kesselmeier, J; Williams, J

    2016-01-22

    The hydroxyl radical (OH) removes most atmospheric pollutants from air. The loss frequency of OH radicals due to the combined effect of all gas-phase OH reactive species is a measureable quantity termed total OH reactivity. Here we present total OH reactivity observations in pristine Amazon rainforest air, as a function of season, time-of-day and height (0-80 m). Total OH reactivity is low during wet (10 s(-1)) and high during dry season (62 s(-1)). Comparison to individually measured trace gases reveals strong variation in unaccounted for OH reactivity, from 5 to 15% missing in wet-season afternoons to mostly unknown (average 79%) during dry season. During dry-season afternoons isoprene, considered the dominant reagent with OH in rainforests, only accounts for ∼20% of the total OH reactivity. Vertical profiles of OH reactivity are shaped by biogenic emissions, photochemistry and turbulent mixing. The rainforest floor was identified as a significant but poorly characterized source of OH reactivity.

  19. Cell Expansion During Directed Differentiation of Stem Cells Toward the Hepatic Lineage

    PubMed Central

    Raju, Ravali; Chau, David; Cho, Dong Seong; Park, Yonsil; Verfaillie, Catherine M.

    2017-01-01

    The differentiation of human pluripotent stem cells toward the hepatocyte lineage can potentially provide an unlimited source of functional hepatocytes for transplantation and extracorporeal bioartificial liver applications. It is anticipated that the quantities of cells needed for these applications will be in the order of 109–1010 cells, because of the size of the liver. An ideal differentiation protocol would be to enable directed differentiation to the hepatocyte lineage with simultaneous cell expansion. We introduced a cell expansion stage after the commitment of human embryonic stem cells to the endodermal lineage, to allow for at least an eightfold increase in cell number, with continuation of cell maturation toward the hepatocyte lineage. The progressive changes in the transcriptome were measured by expression array, and the expression dynamics of certain lineage markers was measured by mass cytometry during the differentiation and expansion process. The findings revealed that while cells were expanding they were also capable of progressing in their differentiation toward the hepatocyte lineage. In addition, our transcriptome, protein and functional studies, including albumin secretion, drug-induced CYP450 expression and urea production, all indicated that the hepatocyte-like cells obtained with or without cell expansion are very similar. This method of simultaneous cell expansion and hepatocyte differentiation should facilitate obtaining large quantities of cells for liver cell applications. PMID:27806669

  20. Lymphocyte apheresis for chimeric antigen receptor T-cell manufacturing in children and young adults with leukemia and neuroblastoma.

    PubMed

    Ceppi, Francesco; Rivers, Julie; Annesley, Colleen; Pinto, Navin; Park, Julie R; Lindgren, Catherine; Mgebroff, Stephanie; Linn, Naomi; Delaney, Meghan; Gardner, Rebecca A

    2018-06-01

    The first step in the production of chimeric antigen receptor T cells is the collection of autologous T cells using apheresis technology. The procedure is technically challenging, because patients often have low leukocyte counts and are heavily pretreated with multiple lines of chemotherapy, marrow transplantation, and/or radiotherapy. Here, we report our experience of collecting T lymphocytes for chimeric antigen receptor T-cell manufacturing in pediatric and young adult patients with leukemia, non-Hodgkin lymphoma, or neuroblastoma. Apheresis procedures were performed on a COBE Spectra machine using the mononuclear cell program, with a collection target of 1 × 10 9 total mononuclear cells per kilogram. Data were collected regarding preapheresis and postapheresis blood counts, apheresis parameters, products, and adverse events. Ninety-nine patients (ages 1.3-25.7 years) and 102 apheresis events were available for analysis. Patients underwent apheresis at a variety of absolute lymphocyte cell counts, with a median absolute lymphocyte count of 944 cells/μL (range, 142-6944 cells/μL). Twenty-two patients (21.6%) had absolute lymphocyte counts less than 500 cells/μL. The mononuclear cell target was obtained in 100% of all apheresis harvests, and chimeric antigen receptor T-cell production was possible from the majority of collections (94%). Mononuclear cell collection efficiency was 65.4%, and T-lymphocyte collection efficiency was 83.4%. Ten patients (9.8%) presented with minor adverse events during the 102 apheresis procedures, with one exception of a severe allergy. Mononuclear cell apheresis for chimeric antigen receptor T-cell therapy is well tolerated and safe, and it is possible to obtain an adequate quantity of CD3+ lymphocytes for chimeric antigen receptor T-cell manufacturing in heavily pretreated patients who have low lymphocyte counts. © 2018 AABB.

  1. Reactor performance of a 750 m(3) anaerobic digestion plant: varied substrate input conditions impacting methanogenic community.

    PubMed

    Wagner, Andreas Otto; Malin, Cornelia; Lins, Philipp; Gstraunthaler, Gudrun; Illmer, Paul

    2014-10-01

    A 750 m(3) anaerobic digester was studied over a half year period including a shift from good reactor performance to a reduced one. Various abiotic parameters like volatile fatty acids (VFA) (formic-, acetic-, propionic-, (iso-)butyric-, (iso-)valeric-, lactic acid), total C, total N, NH4 -N, and total proteins, as well as the organic matter content and dry mass were determined. In addition several process parameters such as temperature, pH, retention time and input of substrate and the concentrations of CH4, H2, CO2 and H2S within the reactor were monitored continuously. The present study aimed at the investigation of the abundance of acetogens and total cell numbers and the microbial methanogenic community as derived from PCR-dHPLC analysis in order to put it into context with the determined abiotic parameters. An influence of substrate quantity on the efficiency of the anaerobic digestion process was found as well as a shift from a hydrogenotrophic in times of good reactor performance towards an acetoclastic dominated methanogenic community in times of reduced reactor performance. After the change in substrate conditions it took the methano-archaeal community about 5-6 weeks to be affected but then changes occurred quickly. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. Potential radiological impact of tornadoes on the safety of Nuclear Fuel Services' West Valley Fuel Reprocessing Plant. Volume I. Tornado effects on head-end cell airflow

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Holloway, L.J.; Andrae, R.W.

    1981-09-01

    This report describes results of a parametric study of the impacts of a tornado-generated depressurization on airflow in the contaminated process cells within the presently inoperative Nuclear Fuel Services fuel reprocessing facility near West Valley, NY. The study involved the following tasks: (1) mathematical modeling of installed ventilation and abnormal exhaust pathways from the cells and prediction of tornado-induced airflows in these pathways; (2) mathematical modeling of individual cell flow characteristics and prediction of in-cell velocities induced by flows from step 1; and (3) evaluation of the results of steps 1 and 2 to determine whether any of the pathwaysmore » investigated have the potential for releasing quantities of radioactively contaminated air from the main process cells. The study has concluded that in the event of a tornado strike, certain pathways from the cells have the potential to release radioactive materials of the atmosphere. Determination of the quantities of radioactive material released from the cells through pathways identified in step 3 is presented in Part II of this report.« less

  3. Why Do Some Batteries Last Longer Than Others?

    NASA Astrophysics Data System (ADS)

    Smith, Michael J.; Vincent, Colin A.

    2002-07-01

    The criteria used by manufacturers to determine the market price of a commercial product are often only indirectly related to what the consumer recognizes as important. This is certainly true of the battery industry; the most expensive battery or cell does not always provide the best service. Even when the electrochemical basis for energy conversion is apparently the same, cells produced by different manufacturers often provide markedly different quantities of energy. In this experiment samples of cathode composite are removed from commercial cells and their electrochemical performance is compared using a test cell and identical discharge conditions. The results confirm that the cell with the most energy does not always have the highest price and suggest that some cell manufacturers may attribute a higher priority to other aspects of performance (power, shelf-life or resistance to abuse, for example), which increase the price without improving the quantity of deliverable energy. The objective of the experiment described in this paper is to provide information that gives the chemically aware consumer a frame of reference for future choice of cells and contributes to an improved understanding of the structure and operational basis of primary cells based on the Leclanché system.

  4. Antibody-based detection of protein phosphorylation status to track the efficacy of novel therapies using nanogram protein quantities from stem cells and cell lines.

    PubMed

    Aspinall-O'Dea, Mark; Pierce, Andrew; Pellicano, Francesca; Williamson, Andrew J; Scott, Mary T; Walker, Michael J; Holyoake, Tessa L; Whetton, Anthony D

    2015-01-01

    This protocol describes a highly reproducible antibody-based method that provides protein level and phosphorylation status information from nanogram quantities of protein cell lysate. Nanocapillary isoelectric focusing (cIEF) combines with UV-activated linking chemistry to detect changes in phosphorylation status. As an example application, we describe how to detect changes in response to tyrosine kinase inhibitors (TKIs) in the phosphorylation status of the adaptor protein CrkL, a major substrate of the oncogenic tyrosine kinase BCR-ABL in chronic myeloid leukemia (CML), using highly enriched CML stem cells and mature cell populations in vitro. This protocol provides a 2.5 pg/nl limit of protein detection (<0.2% of a stem cell sample containing <10(4) cells). Additional assays are described for phosphorylated tyrosine 207 (pTyr207)-CrkL and the protein tyrosine phosphatase PTPRC/CD45; these assays were developed using this protocol and applied to CML patient samples. This method is of high throughput, and it can act as a screen for in vitro cancer stem cell response to drugs and novel agents.

  5. A Perishable Inventory Model with Return

    NASA Astrophysics Data System (ADS)

    Setiawan, S. W.; Lesmono, D.; Limansyah, T.

    2018-04-01

    In this paper, we develop a mathematical model for a perishable inventory with return by assuming deterministic demand and inventory dependent demand. By inventory dependent demand, it means that demand at certain time depends on the available inventory at that time with certain rate. In dealing with perishable items, we should consider deteriorating rate factor that corresponds to the decreasing quality of goods. There are also costs involved in this model such as purchasing, ordering, holding, shortage (backordering) and returning costs. These costs compose the total costs in the model that we want to minimize. In the model we seek for the optimal return time and order quantity. We assume that after some period of time, called return time, perishable items can be returned to the supplier at some returning costs. The supplier will then replace them in the next delivery. Some numerical experiments are given to illustrate our model and sensitivity analysis is performed as well. We found that as the deteriorating rate increases, returning time becomes shorter, the optimal order quantity and total cost increases. When considering the inventory-dependent demand factor, we found that as this factor increases, assuming a certain deteriorating rate, returning time becomes shorter, optimal order quantity becomes larger and the total cost increases.

  6. Optimal ordering and production policy for a recoverable item inventory system with learning effect

    NASA Astrophysics Data System (ADS)

    Tsai, Deng-Maw

    2012-02-01

    This article presents two models for determining an optimal integrated economic order quantity and economic production quantity policy in a recoverable manufacturing environment. The models assume that the unit production time of the recovery process decreases with the increase in total units produced as a result of learning. A fixed proportion of used products are collected from customers and then recovered for reuse. The recovered products are assumed to be in good condition and acceptable to customers. Constant demand can be satisfied by utilising both newly purchased products and recovered products. The aim of this article is to show how to minimise total inventory-related cost. The total cost functions of the two models are derived and two simple search procedures are proposed to determine optimal policy parameters. Numerical examples are provided to illustrate the proposed models. In addition, sensitivity analyses have also been performed and are discussed.

  7. Mass quantity gauging by RF mode analysis

    NASA Technical Reports Server (NTRS)

    Collier, R. S.; Ellerbruch, D.; Cruz, J. E.; Stokes, R. W.; Luft, P. E.; Peterson, R. G.; Hiester, A. E.

    1973-01-01

    Work done to date is reported concerning Radio Frequency Mass Quantity Gauging. Experimental apparatus has been designed and tested which measures the resonant frequencies of a tank in the time domain. These frequencies correspond to the total mass of fluid within the tank. Experimental results are discussed for nitrogen and hydrogen in normal gravity both in the supercritical state and also in the two phase (liquid-gas) region. Theoretical discussions for more general cases are given.

  8. Amount of Televised Alcohol Advertising Exposure and the Quantity of Alcohol Consumed by Youth.

    PubMed

    Naimi, Timothy S; Ross, Craig S; Siegel, Michael B; DeJong, William; Jernigan, David H

    2016-09-01

    Although studies demonstrate that exposure to brand-specific alcohol advertising is associated with an increased likelihood of youth consuming particular brands, the relationship between quantity of brand-specific advertising exposure and quantity of brand-specific consumption has not been firmly established. Using the Alcohol Brand Research Among Underage Drinkers (ABRAND) national sample of 1,031 young drinkers (ages 13-20), this study examined the relationship between their aggregated past-year exposure to advertising (in adstock units, a measure based on gross rating points) for 61 alcohol brands that advertised on the 20 most popular nonsports television programs viewed by underage youth and their aggregated total consumption of those same brands during the past 30 days. Predictive models adjusted for other media exposure, predictors of youth's alcohol consumption, and the consumption of brands not advertised on the 20 shows. For the fully adjusted models, each 100 adstock unit increase in exposure (about 1 SD) was associated with an increase of 5.9 drinks (95% CI [0.9, 11.0 drinks]) consumed during the past 30 days among those with less than 300 units of advertising exposure, and an increase of 55.7 drinks (95% CI [13.9, 97.4 drinks]) among those with 300 or more adstock units of exposure. Among underage youth, the quantity of brand-specific advertising exposure is positively associated with the total quantity of consumption of those advertised brands, even after controlling for the consumption of non-advertised brands. Future research should examine exposure-consumption relationships longitudinally and in other media.

  9. Amount of Televised Alcohol Advertising Exposure and the Quantity of Alcohol Consumed by Youth

    PubMed Central

    Naimi, Timothy S.; Ross, Craig S.; Siegel, Michael B.; DeJong, William; Jernigan, David H.

    2016-01-01

    Objective: Although studies demonstrate that exposure to brand-specific alcohol advertising is associated with an increased likelihood of youth consuming particular brands, the relationship between quantity of brand-specific advertising exposure and quantity of brand-specific consumption has not been firmly established. Method: Using the Alcohol Brand Research Among Underage Drinkers (ABRAND) national sample of 1,031 young drinkers (ages 13–20), this study examined the relationship between their aggregated past-year exposure to advertising (in adstock units, a measure based on gross rating points) for 61 alcohol brands that advertised on the 20 most popular nonsports television programs viewed by underage youth and their aggregated total consumption of those same brands during the past 30 days. Predictive models adjusted for other media exposure, predictors of youth’s alcohol consumption, and the consumption of brands not advertised on the 20 shows. Results: For the fully adjusted models, each 100 adstock unit increase in exposure (about 1 SD) was associated with an increase of 5.9 drinks (95% CI [0.9, 11.0 drinks]) consumed during the past 30 days among those with less than 300 units of advertising exposure, and an increase of 55.7 drinks (95% CI [13.9, 97.4 drinks]) among those with 300 or more adstock units of exposure. Conclusions: Among underage youth, the quantity of brand-specific advertising exposure is positively associated with the total quantity of consumption of those advertised brands, even after controlling for the consumption of non-advertised brands. Future research should examine exposure–consumption relationships longitudinally and in other media. PMID:27588530

  10. Metrics to assess the quantity of antibiotic use in the outpatient setting: a systematic review followed by an international multidisciplinary consensus procedure.

    PubMed

    Versporten, Ann; Gyssens, Inge C; Pulcini, Céline; Monnier, Annelie A; Schouten, Jeroen; Milanic, Romina; Stanic Benic, Mirjana; Tebano, Gianpiero; Le Maréchal, Marion; Zanichelli, Veronica; Huttner, Benedikt; Vlahovic-Palcevski, Vera; Goossens, Herman; Hulscher, Marlies E; Adriaenssens, Niels

    2018-06-01

    The international Innovative Medicines Initiative (IMI) project DRIVE-AB (Driving Reinvestment in Research and Development and Responsible Antibiotic Use) aims to develop a global definition of 'responsible' antibiotic use. To identify consensually validated quantity metrics for antibiotic use in the outpatient setting. First, outpatient quantity metrics (OQMs) were identified by a systematic search of literature and web sites published until 12 December 2014. Identified OQMs were evaluated by a multidisciplinary, international stakeholder panel using a RAND-modified Delphi procedure. Two online questionnaires and a face-to-face meeting between them were conducted to assess OQM relevance for measuring the quantity of antibiotic use on a nine-point Likert scale, to add comments or to propose new metrics. A total of 597 articles were screened, 177 studies met criteria for full-text screening and 138 were finally included. Twenty different OQMs were identified and appraised by 23 stakeholders. During the first survey, 14 OQMs were excluded and 6 qualified for discussion. During the face-to-face meeting, 10 stakeholders retained five OQMs and suggestions were made considering context and combination of metrics. The final set of metrics included defined daily doses, treatments/courses and prescriptions per defined population, treatments/courses and prescriptions per defined number of physician contacts and seasonal variation of total antibiotic use. A small set of consensually validated metrics to assess the quantity of antibiotic use in the outpatient setting was obtained, enabling (inter)national comparisons. The OQMs will help build a global conceptual framework for responsible antibiotic use.

  11. Development and pilot line production of lithium doped silicon solar cells

    NASA Technical Reports Server (NTRS)

    Payne, P. A.

    1972-01-01

    The work performed over the period of September 1971 to August 1972 to develop production processes for fabrication of lithium doped P/N cells is described. The BCl3 diffusion without 02 was selected as the optimum diffusion process for fabrication of lithium doped cells. An 8-2-7 (warm up - deposition - drive-in time in minutes) diffusion schedule at 1055 C was used for the first two lots (300 cells each) delivered to JPL. Cell efficiencies ranged from 11.0 to 13.7% based on an AMO of 135.3 mW/sq cm. These high efficiencies were obtained using from 10 to 40 cells per boron diffusion; increasing the quantity beyond 40 resulted in lower outputs. At this point, the emphasis was placed on investigation of a BCl3 with 02 diffusion. Through evaluation of the effects of diffusion time and temperature, gas flow rates, and desposition plus drive-in vs. continuous deposition and no drive-in cycles, diffusion parameters were determined which produced short circuit currents of 136 + or - 4 mA for ten cells spaced along 12 in. of the diffusion boat. The quantity was increased to 60, 100, and 150 cell diffusions with no more variation in cell short circuit current than observed with 10 cells.

  12. Large-scale preparation of plasmid DNA.

    PubMed

    Heilig, J S; Elbing, K L; Brent, R

    2001-05-01

    Although the need for large quantities of plasmid DNA has diminished as techniques for manipulating small quantities of DNA have improved, occasionally large amounts of high-quality plasmid DNA are desired. This unit describes the preparation of milligram quantities of highly purified plasmid DNA. The first part of the unit describes three methods for preparing crude lysates enriched in plasmid DNA from bacterial cells grown in liquid culture: alkaline lysis, boiling, and Triton lysis. The second part describes four methods for purifying plasmid DNA in such lysates away from contaminating RNA and protein: CsCl/ethidium bromide density gradient centrifugation, polyethylene glycol (PEG) precipitation, anion-exchange chromatography, and size-exclusion chromatography.

  13. Method for providing a low density high strength polyurethane foam

    DOEpatents

    Whinnery, Jr., Leroy L.; Goods, Steven H.; Skala, Dawn M.; Henderson, Craig C.; Keifer, Patrick N.

    2013-06-18

    Disclosed is a method for making a polyurethane closed-cell foam material exhibiting a bulk density below 4 lbs/ft.sup.3 and high strength. The present embodiment uses the reaction product of a modified MDI and a sucrose/glycerine based polyether polyol resin wherein a small measured quantity of the polyol resin is "pre-reacted" with a larger quantity of the isocyanate in a defined ratio such that when the necessary remaining quantity of the polyol resin is added to the "pre-reacted" resin together with a tertiary amine catalyst and water as a blowing agent, the polymerization proceeds slowly enough to provide a stable foam body.

  14. Characteristics and quantities of HIV host cells in human genital tract secretions.

    PubMed

    Politch, Joseph A; Marathe, Jai; Anderson, Deborah J

    2014-12-15

    Human immunodeficiency virus (HIV)-infected leukocytes have been detected in genital secretions from HIV-infected men and women and may play an important role in the sexual transmission of HIV. However, they have been largely overlooked in studies on mechanisms of HIV transmission and in the design and testing of HIV vaccine and microbicide candidates. This article describes the characteristics and quantities of leukocytes in male and female genital secretions under various conditions and also reviews evidence for the involvement of HIV-infected cells in both horizontal and vertical cell-associated HIV transmission. Additional research is needed in this area to better target HIV prevention strategies. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  15. The evolution of tumor metastases during clonal expansion.

    PubMed

    Haeno, Hiroshi; Michor, Franziska

    2010-03-07

    Cancer is a leading cause of morbidity and mortality in many countries. Solid tumors generally initiate at one particular site called the primary tumor, but eventually disseminate and form new colonies in other organs. The development of such metastases greatly diminishes the potential for a cure of patients and is thought to represent the final stage of the multi-stage progression of human cancer. The concept of early metastatic dissemination, however, postulates that cancer cell spread might arise early during the development of a tumor. It is important to know whether metastases are present at diagnosis since this determines treatment strategies and outcome. In this paper, we design a stochastic mathematical model of the evolution of tumor metastases in an expanding cancer cell population. We calculate the probability of metastasis at a given time during tumor evolution, the expected number of metastatic sites, and the total number of cancer cells as well as metastasized cells. Furthermore, we investigate the effect of drug administration and tumor resection on these quantities and predict the survival time of cancer patients. The model presented in this paper allows us to determine the probability and number of metastases at diagnosis and to identify the optimum treatment strategy to maximally prolong survival of cancer patients. 2009 Elsevier Ltd. All rights reserved.

  16. Skin aging in patients with acquired immunodeficiency syndrome.

    PubMed

    de Aquino Favarato, Grace Kelly Naves; da Silva, Aline Cristina Souza; Oliveira, Lívia Ferreira; da Fonseca Ferraz, Mara Lúcia; de Paula Antunes Teixeira, Vicente; Cavellani, Camila Lourencini

    2016-10-01

    To evaluate the histomorphometric skin changes over aging patients with autopsied acquired immunodeficiency syndrome (AIDS). In 29 skin fragments of autopsied elderly (older than 50 years) and nonelderly patients with AIDS, epidermal thickness, the number of layers, the diameter of cells, the percentage of collagen and elastic fibers in the dermis, and the number and morphology of Langerhans cells were assessed. Statistical analysis was performed by SigmaStat 2.03 program. The thickness of the epidermis (92.55 × 158.94 μm), the number of layers (7 × 9 layers), and the diameter of the cells (13.27 × 17.6 μm) were statistically lower among the elderly. The quantity of collagen fibers (9.68 × 14.11%) and elastic fibers (11.89 × 15.31%) was also significantly lower in the elderly. There was a decrease in total (10.61 × 12.38 cel/mm(2)) and an increase in immature Langerhans cells (6.31 × 4.98 cel/mm(2)) in elderly patients with AIDS. The aging of the skin of patients with AIDS is amended in different histomorphometric aspects, the epidermis constituents suffer less pronounced changes in normal aging, and the dermis has more intense changes in elastic fibers and collagen. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. A Multiscale Computational Model Combining a Single Crystal Plasticity Constitutive Model with the Generalized Method of Cells (GMC) for Metallic Polycrystals.

    PubMed

    Ghorbani Moghaddam, Masoud; Achuthan, Ajit; Bednarcyk, Brett A; Arnold, Steven M; Pineda, Evan J

    2016-05-04

    A multiscale computational model is developed for determining the elasto-plastic behavior of polycrystal metals by employing a single crystal plasticity constitutive model that can capture the microstructural scale stress field on a finite element analysis (FEA) framework. The generalized method of cells (GMC) micromechanics model is used for homogenizing the local field quantities. At first, the stand-alone GMC is applied for studying simple material microstructures such as a repeating unit cell (RUC) containing single grain or two grains under uniaxial loading conditions. For verification, the results obtained by the stand-alone GMC are compared to those from an analogous FEA model incorporating the same single crystal plasticity constitutive model. This verification is then extended to samples containing tens to hundreds of grains. The results demonstrate that the GMC homogenization combined with the crystal plasticity constitutive framework is a promising approach for failure analysis of structures as it allows for properly predicting the von Mises stress in the entire RUC, in an average sense, as well as in the local microstructural level, i.e. , each individual grain. Two-three orders of saving in computational cost, at the expense of some accuracy in prediction, especially in the prediction of the components of local tensor field quantities and the quantities near the grain boundaries, was obtained with GMC. Finally, the capability of the developed multiscale model linking FEA and GMC to solve real-life-sized structures is demonstrated by successfully analyzing an engine disc component and determining the microstructural scale details of the field quantities.

  18. A Multiscale Computational Model Combining a Single Crystal Plasticity Constitutive Model with the Generalized Method of Cells (GMC) for Metallic Polycrystals

    PubMed Central

    Ghorbani Moghaddam, Masoud; Achuthan, Ajit; Bednarcyk, Brett A.; Arnold, Steven M.; Pineda, Evan J.

    2016-01-01

    A multiscale computational model is developed for determining the elasto-plastic behavior of polycrystal metals by employing a single crystal plasticity constitutive model that can capture the microstructural scale stress field on a finite element analysis (FEA) framework. The generalized method of cells (GMC) micromechanics model is used for homogenizing the local field quantities. At first, the stand-alone GMC is applied for studying simple material microstructures such as a repeating unit cell (RUC) containing single grain or two grains under uniaxial loading conditions. For verification, the results obtained by the stand-alone GMC are compared to those from an analogous FEA model incorporating the same single crystal plasticity constitutive model. This verification is then extended to samples containing tens to hundreds of grains. The results demonstrate that the GMC homogenization combined with the crystal plasticity constitutive framework is a promising approach for failure analysis of structures as it allows for properly predicting the von Mises stress in the entire RUC, in an average sense, as well as in the local microstructural level, i.e., each individual grain. Two–three orders of saving in computational cost, at the expense of some accuracy in prediction, especially in the prediction of the components of local tensor field quantities and the quantities near the grain boundaries, was obtained with GMC. Finally, the capability of the developed multiscale model linking FEA and GMC to solve real-life-sized structures is demonstrated by successfully analyzing an engine disc component and determining the microstructural scale details of the field quantities. PMID:28773458

  19. A method for quantitative analysis of standard and high-throughput qPCR expression data based on input sample quantity.

    PubMed

    Adamski, Mateusz G; Gumann, Patryk; Baird, Alison E

    2014-01-01

    Over the past decade rapid advances have occurred in the understanding of RNA expression and its regulation. Quantitative polymerase chain reactions (qPCR) have become the gold standard for quantifying gene expression. Microfluidic next generation, high throughput qPCR now permits the detection of transcript copy number in thousands of reactions simultaneously, dramatically increasing the sensitivity over standard qPCR. Here we present a gene expression analysis method applicable to both standard polymerase chain reactions (qPCR) and high throughput qPCR. This technique is adjusted to the input sample quantity (e.g., the number of cells) and is independent of control gene expression. It is efficiency-corrected and with the use of a universal reference sample (commercial complementary DNA (cDNA)) permits the normalization of results between different batches and between different instruments--regardless of potential differences in transcript amplification efficiency. Modifications of the input quantity method include (1) the achievement of absolute quantification and (2) a non-efficiency corrected analysis. When compared to other commonly used algorithms the input quantity method proved to be valid. This method is of particular value for clinical studies of whole blood and circulating leukocytes where cell counts are readily available.

  20. Cyanobacteria and Cyanotoxins Occurrence and Removal from Five High-Risk Conventional Treatment Drinking Water Plants

    PubMed Central

    Szlag, David C.; Sinclair, James L.; Southwell, Benjamin; Westrick, Judy A.

    2015-01-01

    An environmental protection agency EPA expert workshop prioritized three cyanotoxins, microcystins, anatoxin-a, and cylindrospermopsin (MAC), as being important in freshwaters of the United States. This study evaluated the prevalence of potentially toxin producing cyanobacteria cell numbers relative to the presence and quantity of the MAC toxins in the context of this framework. Total and potential toxin producing cyanobacteria cell counts were conducted on weekly raw and finished water samples from utilities located in five US states. An Enzyme-Linked Immunosorbant Assay (ELISA) was used to screen the raw and finished water samples for microcystins. High-pressure liquid chromatography with a photodiode array detector (HPLC/PDA) verified microcystin concentrations and quantified anatoxin-a and cylindrospermopsin concentrations. Four of the five utilities experienced cyanobacterial blooms in their raw water. Raw water samples from three utilities showed detectable levels of microcystins and a fourth utility had detectable levels of both microcystin and cylindrospermopsin. No utilities had detectable concentrations of anatoxin-a. These conventional plants effectively removed the cyanobacterial cells and all finished water samples showed MAC levels below the detection limit by ELISA and HPLC/PDA. PMID:26075379

  1. [The influence of probiotic fermented milk product on colon microbiota, hematological parameters and cell immunity in rats].

    PubMed

    Kuznetsova, G G; Trushina, É N; Muatafina, O K; Cherkashin, A V; Batishcheva, S Iu; Semenikhina, V F; Sheveleva, S A

    2012-01-01

    Influence of probiotic fermented milk product on the intestinal microbiota, hematological parameters and immune status of the experiment in vivo at Wistar rats was studied. It was shown, that entering of probiotic strains of Bifidobacterium bifidum 791, Bifidobacterium longum B-379M and Lactobacillus acidophilus NK1 u Streptococcus thermophilus in composition fermented milk products in the total quantity of 2,1 x 10(7) CFU/ sm3 in digestive tract within three weeks has a positive influence on the resident of colon microbiota. Significant increasing of population levels of Bifidobacterium, Enterobacteriaceae with normal biochemical properties, registered a strong tendency to increase the content of Lactobacteria, which led to a decreasing the number of potential pathogenic transient flora with pathogenic factors. Monitoring of body mass in experimental animals has shown that including of fermented milk product with probiotic strains in diet has a positive influence on the feed uptake. Probiotic properties of the product also have stimulated effect on the immune status of the rat: improvements in cell immunity (increasing the relative amount of T-helper cells, immuneregulatory index) and hematological parameters (increase

  2. Operational Concept Evaluation of Solid Oxide Fuel Cells for Space Vehicle Applications

    NASA Technical Reports Server (NTRS)

    Poast, Kenneth I.

    2011-01-01

    With the end of the Space Shuttle Program, NASA is evaluating many different technologies to support future missions. Green propellants, like liquid methane and liquid oxygen, have potential advantages for some applications. A Lander propelled with LOX/methane engines is one such application. When the total vehicle design and infrastructure are considered, the advantages of the integration of propulsion, heat rejection, life support and power generation become attractive for further evaluation. Scavenged residual propellants from the propulsion tanks could be used to generate needed electric power, heat and water with a Solid Oxide Fuel Cell(SOFC). In-Situ Resource Utilization(ISRU) technologies may also generate quantities of green propellants to refill these tanks and/or supply these fuel cells. Technology demonstration projects such as the Morpheus Lander are currently underway to evaluate the practicality of such designs and operational concepts. Tethered tests are currently in progress on this vertical test bed to evaluate the propulsion and avionics systems. Evaluation of the SOFC seeks to determine the feasibility of using these green propellants to supply power and identify the limits to the integration of this technology into a space vehicle prototype.

  3. Clinical Application of Mesenchymal Stem Cells and Novel Supportive Therapies for Oral Bone Regeneration

    PubMed Central

    O'Valle, Francisco; Lanis, Alejandro; Dohan Ehrenfest, David M.; Wang, Hom-Lay; Galindo-Moreno, Pablo

    2015-01-01

    Bone regeneration is often needed prior to dental implant treatment due to the lack of adequate quantity and quality of the bone after infectious diseases, trauma, tumor, or congenital conditions. In these situations, cell transplantation technologies may help to overcome the limitations of autografts, xenografts, allografts, and alloplastic materials. A database search was conducted to include human clinical trials (randomized or controlled) and case reports/series describing the clinical use of mesenchymal stem cells (MSCs) in the oral cavity for bone regeneration only specifically excluding periodontal regeneration. Additionally, novel advances in related technologies are also described. 190 records were identified. 51 articles were selected for full-text assessment, and only 28 met the inclusion criteria: 9 case series, 10 case reports, and 9 randomized controlled clinical trials. Collectively, they evaluate the use of MSCs in a total of 290 patients in 342 interventions. The current published literature is very diverse in methodology and measurement of outcomes. Moreover, the clinical significance is limited. Therefore, the use of these techniques should be further studied in more challenging clinical scenarios with well-designed and standardized RCTs, potentially in combination with new scaffolding techniques and bioactive molecules to improve the final outcomes. PMID:26064899

  4. [Effects of dexamethasone on the expression of muscarinic receptor mRNA in asthmatic guinea pig airway smooth muscle and eosinophil infiltration in bronchoalveolar lavage fluid].

    PubMed

    Shi, Liang; Luo, Ya-ling; Lai, Wen-yan; Luo, Liang

    2005-08-01

    To investigate the effect of dexamethasone on the expression of muscarinic receptor (MR) mRNA in smooth muscle and infiltration of eosinophils (Eos) in the airway of asthmatic guinea pigs. Thirty healthy guinea pigs were randomized into 3 equal groups, the control group, asthmatic group and dexamethasone therapy group. Asthma was induced in the latter 2 groups with the asthma-inducing agents and received treatments as indicated. Bronchial alveolar lavage fluid(BALF) were collected subsequently from the guinea pigs for examining the total cell number and cell classification, and histopathologic examination of the lung tissue was performed. Semi-quantitative analysis with reverse transcriptional- polymerase chain reaction (RT-PCR) was performed for M(2) and M(3) receptor mRNA in airway smooth muscle. Compared with the control and the asthmatic group, the number of Eos in the BALF of dexamethasone therapy group was significantly lower (P<0.01). In spite of the presence of hyperemia and edema in the lung tissues of the dexamethasone therapy group, Eos infiltration was less severe than that in the asthmatic group. As found by RT-PCR, the quantity of M(2) receptor mRNA in the airway smooth muscle of the dexamethasone therapy group was significantly higher than those in both the control and asthmatic groups (P<0.01), and the quantity of M(3) receptor mRNA in the airway smooth muscle of dexamethasone therapy group was significantly higher than that in the asthmatic group, but did not significantly differ from that in the control group. The quantities of M(2) and M(3) receptor mRNAs in the control group were both significantly higher than that in asthmatic group (P<0.01). The expression of M(2) receptor is increased in antigen- challenged guinea pigs, and that of M(3) receptor decreased. Dexamethasone can treat asthma by inhibiting inflammatory action involving Eos infiltration, regulating the expressions of M(2) and M(3) receptors and restoring the function of M(2) receptor.

  5. Effect of genetic and nongenetic factors on chemical composition of individual milk samples from dromedary camels (Camelus dromedarius) under intensive management.

    PubMed

    Nagy, P; Fábri, Zs N; Varga, L; Reiczigel, J; Juhász, J

    2017-11-01

    The aims of the present study were to monitor the changes in gross chemical composition of individual dromedary camel milk over a 5-yr period, to provide reference values, and to determine the effect of genetic and nongenetic factors influencing camel milk composition under intensive management. A total of 1,528 lactating dromedary camels were included in the study. Animals were fed a constant diet and were milked twice a day in a herringbone parlor. Milk samples were collected at monthly intervals using a sampling device and then fat, protein, lactose, total solids (TS), and solids-nonfat (SNF) concentrations of raw camel milk were determined with an automatic milk analyzer. For each milk sample, production parameters were recorded and quantities (grams) of milk constituents were calculated. The overall mean quantity and fat, protein, lactose, SNF, and TS concentrations of the morning milk were 4.0 kg, 2.58%, 2.95%, 4.19%, 8.08%, and 10.46%, respectively. Milk quantity showed a positive correlation with lactose and a negative correlation with all other components. Parity exerted a strong effect on all milk parameters. Primiparous dromedaries (n = 60) produced less milk with higher concentrations of components than did multiparous animals (n = 1,468). Milk composition varied among the 7 breeds tested, but none of the genotypes was found to be superior to the others in this respect. We detected a significant, yet small calf sex-biased difference in milk yield and composition. Stage of lactation and season strongly influenced milk yield and all milk components. We also found a significant interaction between month postpartum (mPP) and month of the year. The concentration of all milk components decreased from 1 to 5 mPP. Later, lactose concentration and quantity continued to decrease parallel with decreasing milk production. The concentration of other components showed a temporary increase in mid lactation, from 6 to 11 mPP, and in late lactation, from 18 to 23 mPP. Mean fat, protein, SNF, and TS concentrations showed a high seasonal variation (9.5 to 28.7%), with the lowest and highest values being measured during summer and winter, respectively. This seasonal variation was independent of nutrition and may reflect an endogenous circannual rhythm. We observed a noticeable variation among years. Dromedary camels could provide a useful in vivo model to study the homeorhetic regulation of mammary cell function by endogenous and environmental factors. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  6. 17 CFR 32.5 - Disclosure.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... transactions being offered including: (i) The duration of the commodity options being offered and the total quantity and quality of the commodities which may be purchased or sold upon exercise of the options being... PERSON SHOULD NOT PURCHASE A COMMODITY OPTION UNLESS HE IS PREPARED TO SUSTAIN A TOTAL LOSS OF THE...

  7. 7 CFR 457.123 - Almond crop insurance provisions.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... pattern. Meat pounds. The total pounds of almond meats (whole, chipped and broken, and in-shell meats). In-shell almonds will be converted to meat pounds in accordance with FCIC approved procedures. Production guarantee (per acre). The quantity of almonds (total meat pounds per acre) determined by multiplying the...

  8. 7 CFR 457.123 - Almond crop insurance provisions.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... pattern. Meat pounds. The total pounds of almond meats (whole, chipped and broken, and in-shell meats). In-shell almonds will be converted to meat pounds in accordance with FCIC approved procedures. Production guarantee (per acre). The quantity of almonds (total meat pounds per acre) determined by multiplying the...

  9. 7 CFR 457.123 - Almond crop insurance provisions.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... pattern. Meat pounds. The total pounds of almond meats (whole, chipped and broken, and in-shell meats). In-shell almonds will be converted to meat pounds in accordance with FCIC approved procedures. Production guarantee (per acre). The quantity of almonds (total meat pounds per acre) determined by multiplying the...

  10. In-depth Characterization of a TCR-specific Tracer for Sensitive Detection of Tumor-directed Transgenic T Cells by Immuno-PET.

    PubMed

    Yusufi, Nahid; Mall, Sabine; Bianchi, Henrique de Oliveira; Steiger, Katja; Reder, Sybille; Klar, Richard; Audehm, Stefan; Mustafa, Mona; Nekolla, Stephan; Peschel, Christian; Schwaiger, Markus; Krackhardt, Angela M; D'Alessandria, Calogero

    2017-01-01

    A number of different technologies have been developed to monitor in vivo the distribution of gene-modified T cells used in immunotherapy. Nevertheless, in-depth characterization of novel approaches with respect to sensitivity and clinical applicability are so far missing. We have previously described a novel method to track engineered human T cells in tumors using 89 Zr-Df-aTCRmu-F(ab') 2 targeting the murinized part of the TCR beta domain (TCRmu) of a transgenic TCR. Here, we performed an in-depth in vitro characterization of the tracer in terms of antigen affinity, immunoreactivity, influence on T-cell functionality and stability in vitro and in vivo . Of particular interest, we have developed diverse experimental settings to quantify TCR-transgenic T cells in vivo . Local application of 89 Zr-Df-aTCRmu-F(ab') 2 -labeled T cells in a spot-assay revealed signal detection down to approximately 1.8x10 4 cells. In a more clinically relevant model, NSG mice were intravenously injected with different numbers of transgenic T cells, followed by injection of the 89 Zr-Df-aTCRmu-F(ab') 2 tracer, PET/CT imaging and subsequent ex vivo T-cell quantification in the tumor. Using this setting, we defined a comparable detection limit of 1.0x10 4 T cells. PET signals correlated well to total numbers of transgenic T cells detected ex vivo independently of the engraftment rates observed in different individual experiments. Thus, these findings confirm the high sensitivity of our novel PET/CT T-cell tracking method and provide critical information about the quantity of transgenic T cells in the tumor environment suggesting our technology being highly suitable for further clinical translation.

  11. MANIFESTATIONS OF INJURY IN YEAST CELLS EXPOSED TO SUBZERO TEMPERATURES II.

    PubMed Central

    Mazur, Peter

    1961-01-01

    Mazur, Peter (Oak Ridge National Laboratory, Oak Ridge, Tenn.). Manifestations of injury in yeast cells exposed to subzero temperatures. II. Changes in specific gravity and in the concentration and quantity of cell solids. J. Bacteriol. 82:673–684. 1961.—It has previously been established that subjecting cells of Saccharomyces cerevisiae to rapid cooling to −30 C results in cell death and in certain morphological alterations. The alterations consisted of the loss of the central vacuole and a 50% decrease in volume. The present experiments were concerned with determining whether the volume decrease was the result of the loss of water alone or of water plus cellular solutes. The density of the “frozenthawed” cells was found to increase from 1.14 to 1.25 g/cm3 on the basis of measurements of the sedimentation rate of the cells. Interferometric and refractometric measurements indicated, furthermore, that the concentration of cell solids increased from 20 to 28%, whereas the total mass of cell solids decreased from 25 to 17 μμg/cell. The decrease in cell volume was thus shown to be the result of loss of solution from the cells, a solution containing 11 to 16% solids. Measurements of the rate of dialysis suggested that most or all of these solids had a molecular weight below 600. The findings are consistent with the view that low-temperature exposure destroyed the vacuolar membrane and sufficiently damaged the permeability barriers of the cell to permit escape of low molecular weight compounds. The damage was present a few seconds after thawing, and may, therefore, have been a direct result of intracellular ice crystals which, on the basis of previous studies, are believed to be responsible for death from low-temperature exposure. PMID:14471819

  12. Drude Weight, Meissner Weight, Rotational Inertia of Bosonic Superfluids: How Are They Distinguished?

    NASA Astrophysics Data System (ADS)

    Hetényi, Balázs

    2014-03-01

    The Drude weight, the quantity which distinguishes metals from insulators, is proportional to the second derivative of the ground state energy with respect to a flux at zero flux. The same expression also appears in the definition of the Meissner weight, the quantity which indicates superconductivity, as well as in the definition of non-classical rotational inertia of bosonic superfluids. It is shown that the difference between these quantities depends on the interpretation of the average momentum term, which can be understood as the expectation value of the total momentum (Drude weight), the sum of the expectation values of single momenta (rotational inertia of a superfluid), or the sum over expectation values of momentum pairs (Meissner weight). This distinction appears naturally when the current from which the particular transport quantity is derived is cast in terms of shift operators.

  13. Analysis of Radiation-Induced Chromosomal Aberrations on a Cell-by-Cell Basis after Alpha-Particle Microbeam Irradiation: Experimental Data and Simulations.

    PubMed

    Testa, Antonella; Ballarini, Francesca; Giesen, Ulrich; Gil, Octávia Monteiro; Carante, Mario P; Tello, John; Langner, Frank; Rabus, Hans; Palma, Valentina; Pinto, Massimo; Patrono, Clarice

    2018-06-01

    There is a continued need for further clarification of various aspects of radiation-induced chromosomal aberration, including its correlation with radiation track structure. As part of the EMRP joint research project, Biologically Weighted Quantities in Radiotherapy (BioQuaRT), we performed experimental and theoretical analyses on chromosomal aberrations in Chinese hamster ovary cells (CHO-K1) exposed to α particles with final energies of 5.5 and 17.8 MeV (absorbed doses: ∼2.3 Gy and ∼1.9 Gy, respectively), which were generated by the microbeam at the Physikalisch-Technische Bundesanstalt (PTB) in Braunschweig, Germany. In line with the differences in linear energy transfer (approximately 85 keV/μm for 5.5 MeV and 36 keV/μm for 17.8 MeV α particles), the 5.5 MeV α particles were more effective than the 17.8 MeV α particles, both in terms of the percentage of aberrant cells (57% vs. 33%) and aberration frequency. The yield of total aberrations increased by a factor of ∼2, although the increase in dicentrics plus centric rings was less pronounced than in acentric fragments. The experimental data were compared with Monte Carlo simulations based on the BIophysical ANalysis of Cell death and chromosomal Aberrations model (BIANCA). This comparison allowed interpretation of the results in terms of critical DNA damage [cluster lesions (CLs)]. More specifically, the higher aberration yields observed for the 5.5 MeV α particles were explained by taking into account that, although the nucleus was traversed by fewer particles (nominally, 11 vs. 25), each particle was much more effective (by a factor of ∼3) at inducing CLs. This led to an increased yield of CLs per cell (by a factor of ∼1.4), consistent with the increased yield of total aberrations observed in the experiments.

  14. A Search Algorithm for Determination of Economic Order Quantity in a Two-Level Supply Chain System with Transportation Cost

    NASA Astrophysics Data System (ADS)

    Pirayesh Neghab, Mohammadali; Haji, Rasoul

    This study considers a two-level supply chain system consisting of one warehouse and a number of identical retailers. In this system, we incorporate transportation costs into inventory replenishment decisions. The transportation cost contains a fixed cost and a variable cost. We assume that the demand rate at each retailer is known and the demand is confined to a single item. First, we derive the total cost which is the sum of the holding and ordering cost at the warehouse and retailers as well as the transportation cost from the warehouse to retailers. Then, we propose a search algorithm to find the economic order quantities for the warehouse and retailers which minimize the total cost.

  15. Collective synthesis of natural products by means of organocascade catalysis

    PubMed Central

    Jones, Spencer B.; Simmons, Bryon; Mastracchio, Anthony; MacMillan, David W. C.

    2012-01-01

    Organic chemists are now able to synthesize small quantities of almost any known natural product, given sufficient time, resources and effort. However, translation of the academic successes in total synthesis to the large-scale construction of complex natural products and the development of large collections of biologically relevant molecules present significant challenges to synthetic chemists. Here we show that the application of two nature-inspired techniques, namely organocascade catalysis and collective natural product synthesis, can facilitate the preparation of useful quantities of a range of structurally diverse natural products from a common molecular scaffold. The power of this concept has been demonstrated through the expedient, asymmetric total syntheses of six well-known alkaloid natural products: strychnine, aspidospermidine, vincadifformine, akuammicine, kopsanone and kopsinine. PMID:21753848

  16. [Comparison of green coffee beans volatiles chemical composition of Hainan main area].

    PubMed

    Hu, Rong-Suo; Chu, Zhong; Gu, Feng-Lin; Lu, Min-Quan; Lu, Shao-Fang; Wu, Gui-Ping; Tan, Le-He

    2013-02-01

    Chemical component of Hainan green coffee beans was analyzed with solid phase microextraction-gas chromatography-mass spectrometry, and the discrepancy between two green coffee beans was differentiated through the spectrum database retrieval and retention index of compound characterization. The experimental results show that: the chemical composition of Wanning coffee beans and Chengmai coffee beans is basically the same. The quantity of analyzed compound in Wanning area coffee is 91, and in Chengmai area coffee is 106, the quantity of the same compound is 66, and the percent of the same component is 75.52%. The same compounds accounted for 89.86% of the total content of Wanning area coffee, and accounted for 85.70% of the total content of Chengmai area coffee.

  17. Semi-automated extraction and characterization of Stromal Vascular Fraction using a new medical device.

    PubMed

    Hanke, Alexander; Prantl, Lukas; Wenzel, Carina; Nerlich, Michael; Brockhoff, Gero; Loibl, Markus; Gehmert, Sebastian

    2016-01-01

    The stem cell rich Stromal Vascular Fraction (SVF) can be harvested by processing lipo-aspirate or fat tissue with an enzymatic digestion followed by centrifugation. To date neither a standardised extraction method for SVF nor a generally admitted protocol for cell application in patients exists. A novel commercially available semi-automated device for the extraction of SVF promises sterility, consistent results and usability in the clinical routine. The aim of this work was to compare the quantity and quality of the SVF between the new system and an established manual laboratory method. SVF was extracted from lipo-aspirate both by a prototype of the semi-automated UNiStation™ (NeoGenesis, Seoul, Korea) and by hand preparation with common laboratory equipment. Cell composition of the SVF was characterized by multi-parametric flow-cytometry (FACSCanto-II, BD Biosciences). The total cell number (quantity) of the SVF was determined as well the percentage of cells expressing the stem cell marker CD34, the leucocyte marker CD45 and the marker CD271 for highly proliferative stem cells (quality). Lipo-aspirate obtained from six patients was processed with both the novel device (d) and the hand preparation (h) which always resulted in a macroscopically visible SVF. However, there was a tendency of a fewer cell yield per gram of used lipo-aspirate with the device (d: 1.1×105±1.1×105 vs. h: 2.0×105±1.7×105; p = 0.06). Noteworthy, the percentage of CD34+ cells was significantly lower when using the device (d: 57.3% ±23.8% vs. h: 74.1% ±13.4%; p = 0.02) and CD45+ leukocyte counts tend to be higher when compared to the hand preparation (d: 20.7% ±15.8% vs. h: 9.8% ±7.1%; p = 0.07). The percentage of highly proliferative CD271+ cells was similar for both methods (d:12.9% ±9.6% vs. h: 13.4% ±11.6%; p = 0.74) and no differences were found for double positive cells of CD34+/CD45+ (d: 5.9% ±1.7% vs. h: 1.7% ±1.1%; p = 0.13), CD34+/CD271+ (d: 24.1% ±12.0% vs. h: 14.2% ±8.5%; p = 0.07). The semi-automated closed system provides a considerable amount of sterile SVF with high reproducibility. Furthermore, the SVF extracted by both methods showed a similar cell composition which is in accordance with the data from literature. This semi-automated device offers an opportunity to take research and application of the SVF one step further to the clinic.

  18. Gigantism in a bacterium, Epulopiscium fishelsoni, correlates with complex patterns in arrangement, quantity, and segregation of DNA.

    PubMed

    Bresler, V; Montgomery, W L; Fishelson, L; Pollak, P E

    1998-11-01

    Epulopiscium fishelsoni, gut symbiont of the brown surgeonfish (Acanthurus nigrofuscus) in the Red Sea, attains a larger size than any other eubacterium, varies 10- to 20-fold in length (and >2, 000-fold in volume), and undergoes a complex daily life cycle. In early morning, nucleoids contain highly condensed DNA in elongate, chromosome-like structures which are physically separated from the general cytoplasm. Cell division involves production of two (rarely three) nucleoids within a cell, deposition of cell walls around expanded nucleoids, and emergence of daughter cells from the parent cell. Fluorescence measurements of DNA, RNA, and other cell components indicate the following. DNA quantity is proportional to cell volume over cell lengths of approximately 30 micrometers to >500 micrometers. For cells of a given size, nucleoids of cells with two nucleoids (binucleoid) contain approximately equal amounts of DNA. And each nucleoid of a binucleoid cell contains one-half the DNA of the single nucleoid in a uninucleoid cell of the same size. The life cycle involves approximately equal subdivision of DNA among daughter cells, formation of apical caps of condensed DNA from previously decondensed and diffusely distributed DNA, and "pinching" of DNA near the middle of the cell in the absence of new wall formation. Mechanisms underlying these patterns remain unclear, but formation of daughter nucleoids and cells occurs both during diurnal periods of host feeding and bacterial cell growth and during nocturnal periods of host inactivity when mean bacterial cell size declines.

  19. A modified protocol for RNA extraction from different peach tissues suitable for gene isolation and real-time PCR analysis.

    PubMed

    Tong, Zhaoguo; Qu, Shenchun; Zhang, Jiyu; Wang, Fei; Tao, Jianmin; Gao, Zhihong; Zhang, Zhen

    2012-03-01

    RNA extraction is the first step in the study of gene isolation and expression. However, it is difficult to extract high quantity and quality RNA from tissues containing large quantities of polysaccharides and polyphenols. Peach (Prunus persica), in addition to containing high levels of polysaccharides and polyphenols, is a challenging starting material for RNA isolation using a single method because of different amounts of those substances in diverse tissues. Based on three reported methods, we developed a modified RNA isolation protocol to solve this problem, leading to high quality and quantity of total RNA from peach mesocarp tissues of fruits which were sampled from all developmental stages and different storage periods, as well as from other tissues including flowers, leaves, stems, and roots. With our modified method, 28-650 μg of total RNA was routinely obtained from per gram of fresh material, gave at least a 1.16-fold improvement by compared with those isolated by other seven methods. The RNA extracts were successfully used in downstream applications such as RT-PCR, RACE, and real-time PCR.

  20. Vendor-buyer inventory models with trade credit financing under both non-cooperative and integrated environments

    NASA Astrophysics Data System (ADS)

    Teng, Jinn-Tsair; Chang, Chun-Tao; Chern, Maw-Sheng

    2012-11-01

    Most researchers studied vendor-buyer supply chain inventory policies only from the perspective of an integrated model, which provides us the best cooperative solution. However, in reality, not many vendors and buyers are wholly integrated. Hence, it is necessary to study the optimal policies not only under an integrated environment but also under a non-cooperative environment. In this article, we develop a supply chain vendor-buyer inventory model with trade credit financing linked to order quantity. We then study the optimal policies for both the vendor and the buyer under a non-cooperative environment first, and then under a cooperative integrated situation. Further, we provide some numerical examples to illustrate the theoretical results, compare the differences between these two distinct solutions, and obtain some managerial insights. For example, in a cooperative environment, to reduce the total cost for both parties, the vendor should either provide a simple permissible delay without order quantity restriction or offer a long permissible delay linked order quantity. By contrast, in a non-cooperative environment, the vendor should provide a short permissible delay to reduce its total cost.

  1. Current databases on biological variation: pros, cons and progress.

    PubMed

    Ricós, C; Alvarez, V; Cava, F; García-Lario, J V; Hernández, A; Jiménez, C V; Minchinela, J; Perich, C; Simón, M

    1999-11-01

    A database with reliable information to derive definitive analytical quality specifications for a large number of clinical laboratory tests was prepared in this work. This was achieved by comparing and correlating descriptive data and relevant observations with the biological variation information, an approach that had not been used in the previous efforts of this type. The material compiled in the database was obtained from published articles referenced in BIOS, CURRENT CONTENTS, EMBASE and MEDLINE using "biological variation & laboratory medicine" as key words, as well as books and doctoral theses provided by their authors. The database covers 316 quantities and reviews 191 articles, fewer than 10 of which had to be rejected. The within- and between-subject coefficients of variation and the subsequent desirable quality specifications for precision, bias and total error for all the quantities accepted are presented. Sex-related stratification of results was justified for only four quantities and, in these cases, quality specifications were derived from the group with lower within-subject variation. For certain quantities, biological variation in pathological states was higher than in the healthy state. In these cases, quality specifications were derived only from the healthy population (most stringent). Several quantities (particularly hormones) have been treated in very few articles and the results found are highly discrepant. Therefore, professionals in laboratory medicine should be strongly encouraged to study the quantities for which results are discrepant, the 90 quantities described in only one paper and the numerous quantities that have not been the subject of study.

  2. Proteolytic Regulation of the Intestinal Epithelial Barrier: Mechanisms and Interventions

    DTIC Science & Technology

    2014-09-01

    DSS protocol to evaluate molecular markers of acute inflammation in the subepithelial lamina propria, including quantity and nature of immune cell...will be investigated by immunostaining of colonic segments for Ki-67, a nuclear protein preferentially expressed during active phases of the cell

  3. RESEARCH ON CELL WALL CYTOCHEMISTRY OF SELECTED FUNGI.

    DTIC Science & Technology

    Cytochemical analyses of the four imperfect fungi, Cylindrocephalum sp., Curvularia lunata, Aspergillus oryzae , and Gliocladium deliquescens showed...forming plastids increase in number by direct division and the quantities of chitin at their surfaces, in a given cell, may be equal to or greater than the

  4. Light-stimulated cell expansion in bean (Phaseolus vulgaris L.) leaves. II. Quantity and quality of light required

    NASA Technical Reports Server (NTRS)

    Van Volkenburgh, E.; Cleland, R. E.; Watanabe, M.

    1990-01-01

    The quantity and quality of light required for light-stimulated cell expansion in leaves of Phaseolus vulgaris L. have been determined. Seedlings were grown in dim red light (RL; 4 micromoles photons m-2 s-1) until cell division in the primary leaves was completed, then excised discs were incubated in 10 mM sucrose plus 10 mM KCl in a variety of light treatments. The growth response of discs exposed to continuous white light (WL) for 16 h was saturated at 100 micromoles m-2 s-1, and did not show reciprocity. Extensive, but not continuous, illumination was needed for maximal growth. The wavelength dependence of disc expansion was determined from fluence-response curves obtained from 380 to 730 nm provided by the Okazaki Large Spectrograph. Blue (BL; 460 nm) and red light (RL; 660 nm) were most effective in promoting leaf cell growth, both in photosynthetically active and inhibited leaf discs. Far-red light (FR; 730 nm) reduced the effectiveness of RL, but not BL, indicating that phytochrome and a separate blue-light receptor mediate expansion of leaf cells.

  5. Microscopic Analysis and Quality Assessment of Induced Sputum From Children With Pneumonia in the PERCH Study.

    PubMed

    Murdoch, David R; Morpeth, Susan C; Hammitt, Laura L; Driscoll, Amanda J; Watson, Nora L; Baggett, Henry C; Brooks, W Abdullah; Deloria Knoll, Maria; Feikin, Daniel R; Kotloff, Karen L; Levine, Orin S; Madhi, Shabir A; O'Brien, Katherine L; Scott, J Anthony G; Thea, Donald M; Ahmed, Dilruba; Awori, Juliet O; DeLuca, Andrea N; Ebruke, Bernard E; Higdon, Melissa M; Jorakate, Possawat; Karron, Ruth A; Kazungu, Sidi; Kwenda, Geoffrey; Hossain, Lokman; Makprasert, Sirirat; Moore, David P; Mudau, Azwifarwi; Mwaba, John; Panchalingam, Sandra; Park, Daniel E; Prosperi, Christine; Salaudeen, Rasheed; Toure, Aliou; Zeger, Scott L; Howie, Stephen R C

    2017-06-15

    It is standard practice for laboratories to assess the cellular quality of expectorated sputum specimens to check that they originated from the lower respiratory tract. The presence of low numbers of squamous epithelial cells (SECs) and high numbers of polymorphonuclear (PMN) cells are regarded as indicative of a lower respiratory tract specimen. However, these quality ratings have never been evaluated for induced sputum specimens from children with suspected pneumonia. We evaluated induced sputum Gram stain smears and cultures from hospitalized children aged 1-59 months enrolled in a large study of community-acquired pneumonia. We hypothesized that a specimen representative of the lower respiratory tract will contain smaller quantities of oropharyngeal flora and be more likely to have a predominance of potential pathogens compared to a specimen containing mainly saliva. The prevalence of potential pathogens cultured from induced sputum specimens and quantity of oropharyngeal flora were compared for different quantities of SECs and PMNs. Of 3772 induced sputum specimens, 2608 (69%) had <10 SECs per low-power field (LPF) and 2350 (62%) had >25 PMNs per LPF, measures traditionally associated with specimens from the lower respiratory tract in adults. Using isolation of low quantities of oropharyngeal flora and higher prevalence of potential pathogens as markers of higher quality, <10 SECs per LPF (but not >25 PMNs per LPF) was the microscopic variable most associated with high quality of induced sputum. Quantity of SECs may be a useful quality measure of induced sputum from young children with pneumonia. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America.

  6. Exploiting active subspaces to quantify uncertainty in the numerical simulation of the HyShot II scramjet

    NASA Astrophysics Data System (ADS)

    Constantine, P. G.; Emory, M.; Larsson, J.; Iaccarino, G.

    2015-12-01

    We present a computational analysis of the reactive flow in a hypersonic scramjet engine with focus on effects of uncertainties in the operating conditions. We employ a novel methodology based on active subspaces to characterize the effects of the input uncertainty on the scramjet performance. The active subspace identifies one-dimensional structure in the map from simulation inputs to quantity of interest that allows us to reparameterize the operating conditions; instead of seven physical parameters, we can use a single derived active variable. This dimension reduction enables otherwise infeasible uncertainty quantification, considering the simulation cost of roughly 9500 CPU-hours per run. For two values of the fuel injection rate, we use a total of 68 simulations to (i) identify the parameters that contribute the most to the variation in the output quantity of interest, (ii) estimate upper and lower bounds on the quantity of interest, (iii) classify sets of operating conditions as safe or unsafe corresponding to a threshold on the output quantity of interest, and (iv) estimate a cumulative distribution function for the quantity of interest.

  7. Characterization and differentiation of human embryonic stem cells.

    PubMed

    Carpenter, M K; Rosler, E; Rao, M S

    2003-01-01

    Cell replacement therapies have been limited by the availability of sufficient quantities of cells for transplantation. Human ES (hES) cell lines have recently been generated by several laboratories. When maintained for over 1 year in vitro, they remain karyotypically and phenotypically stable and may therefore provide an excellent source material for cell therapies. Currently, data is available for 26 hES cell lines. Although limited characterization has been performed on most of these lines, there are remarkable similarities in expression of markers. hES cell lines derived in different laboratories show similar expression profiles of surface markers, including SSEA-4, Tra-1-60, and Tra-1-81. In addition, markers associated with pluripotent cells such as OCT-4 are expressed at in all cell lines tested. These cells express high levels of telomerase and appear to have indefinite growth potential. The generation of the large quantities of cells necessary for cell replacement therapies will require a cell population which is stable over long term culture. We have characterized the properties of multiple hES cell lines that have been maintained in culture for extended periods. Quantitative analyses demonstrate that all of the cell lines examined show consistent marker expression and retain a normal karyotype after long-term culture. hES cells have been differentiated into the derivatives of all three germ layers. Specifically this includes cardiomyocytes, neural cells, hepatocyte-like cells, endothelial cells and hematopoietic progenitor cells. These data demonstrating the karyotypic and phenotypic stability of hES cells and their extensive differentiative capacity indicate that they may be an appropriate source of cells for multiple regenerative medicine applications.

  8. How the great apes (Pan troglodytes, Pongo pygmaeus, Pan paniscus, Gorilla gorilla) perform on the reversed reward contingency task II: transfer to new quantities, long-term retention, and the impact of quantity ratios.

    PubMed

    Uher, Jana; Call, Josep

    2008-05-01

    We tested 6 chimpanzees (Pan troglodytes), 3 orangutans (Pongo pygmaeus), 4 bonobos (Pan paniscus), and 2 gorillas (Gorilla gorilla) in the reversed reward contingency task. Individuals were presented with pairs of quantities ranging between 0 and 6 food items. Prior to testing, some experienced apes had solved this task using 2 quantities while others were totally naïve. Experienced apes transferred their ability to multiple-novel pairs after 6 to 19 months had elapsed since their initial testing. Two out of 6 naïve apes (1 chimpanzee, 1 bonobo) solved the task--a proportion comparable to that of a previous study using 2 pairs of quantities. Their acquisition speed was also comparable to the successful subjects from that study. The ratio between quantities explained a large portion of the variance but affected naïve and experienced individuals differently. For smaller ratios, naïve individuals were well below 50% correct and experienced ones were well above 50%, yet both groups tended to converge toward 50% for larger ratios. Thus, some apes require no procedural modifications to overcome their strong bias for selecting the larger of 2 quantities. PsycINFO Database Record (c) 2008 APA, all rights reserved.

  9. Relative quantity judgments in the beluga whale (Delphinapterus leucas) and the bottlenose dolphin (Tursiops truncatus).

    PubMed

    Abramson, José Z; Hernández-Lloreda, Victoria; Call, Josep; Colmenares, Fernando

    2013-06-01

    Numerous studies have documented the ability of many species to make relative quantity judgments using an analogue magnitude system. We investigated whether one beluga whale, Delphinapterus leucas, and three bottlenose dolphins, Tursiops truncatus, were capable of selecting the larger of two sets of quantities, and analyzed if their performance matched predictions from the object file model versus the analog accumulator model. In Experiment 1, the two sets were presented simultaneously, under water, and they were visually (condition 1) or echoically (condition 2) available at the time of choice. In experiment 2, the two sets were presented above the water, successively (condition 1) or sequentially, item-by-item (condition 2), so that they were not visually available at the time of choice (condition 1) or at any time throughout the experiment (condition 2). We analyzed the effect of the ratio between quantities, the difference between quantities, and the total number of items presented on the subjects' choices. All subjects selected the larger of the two sets of quantities above chance levels in all conditions. However, unlike most previous studies, the subjects' choices did not match the predictions from the accumulator model. Whether these findings reflect interspecies differences in the mechanisms which underpin relative quantity judgments remains to be determined. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Fluconazole impacts the extracellular matrix of fluconazole-susceptible and -resistant Candida albicans and Candida glabrata biofilms.

    PubMed

    Panariello, Beatriz Helena Dias; Klein, Marlise I; Mima, Ewerton Garcia De Oliveira; Pavarina, Ana Cláudia

    2018-01-01

    Background : Fluconazole (FLZ) is a drug commonly used for the treatment of Candida infections. However, β-glucans in the extracellular matrices (ECMs) hinder FLZ penetration into Candida biofilms, while extracellular DNA (eDNA) contributes to the biofilm architecture and resistance. Methods : This study characterized biofilms of FLZ-sensitive (S) and -resistant (R) Candida albicans and Candida glabrata in the presence or absence of FLZ focusing on the ECM traits. Biofilms of C. albicans American Type Culture Collection (ATCC) 90028 (CaS), C. albicans ATCC 96901 (CaR), C. glabrata ATCC 2001 (CgS), and C. glabrata ATCC 200918 (CgR) were grown in RPMI medium with or without FLZ at 5× the minimum inhibitory concentration (37°C/48 h). Biofilms were assessed by colony-forming unit (CFU)/mL, biomass, and ECM components (alkali-soluble polysaccharides [ASP], water-soluble polysaccharides [WSP], eDNA, and proteins). Scanning electron microscopy (SEM) was also performed. Data were analyzed by parametric and nonparametric tests ( α   =  0.05). Results : In biofilms, FLZ reduced the CFU/mL of all strains ( p  < 0.001), except for CaS ( p  = 0.937). However, the ASP quantity in CaS was significantly reduced by FLZ ( p  = 0.034), while the drug had no effect on the ASP levels in other strains ( p  > 0.05). Total biomasses and WSP were significantly reduced by FLZ in the ECM of all yeasts ( p  < 0.001), but levels of eDNA and proteins were unaffected ( p  > 0.05). FLZ affected the cell morphology and biofilm structure by hindering hyphae formation in CaS and CaR biofilms, by decreasing the number of cells in CgS and CgR biofilms, and by yielding sparsely spaced cell agglomerates on the substrate. Conclusion : FLZ impacts biofilms of C. albicans and C. glabrata as evident by reduced biomass. This reduced biomass coincided with lowered cell numbers and quantity of WSPs. Hyphal production by C. albicans was also reduced.

  11. Beta-Adrenergic Receptor Population is Up-Regulated by Increased Cyclic Amp Concentration in Chicken Skeletal Muscle Cells in Culture

    NASA Technical Reports Server (NTRS)

    Young, Ronald B.; Bridge, Kristin Y.; Vaughn, Jeffrey R.

    1999-01-01

    Skeletal muscle hypertrophy is promoted in vivo by administration of beta-drenergic receptor (bAR) agonists. Chicken skeletal muscle cells were treated with 1 (mu)M isoproterenol, a strong bAR agonist, between days 7 and 10 in culture. bAR population increased by approximately 40% during this treatment; however, the ability of the cells to synthesize cyclic AMP (cAMP) was diminished by two-fold. The quantity of myosin heavy chain (MHC) was not affected. To understand further the relationship between intracellular cAMP levels, bAR population, and muscle protein accumulation, intracellular cAMP levels were artificially elevated by treatment with 0-10 uM forskolin for up to three days. The basal concentration of CAMP in forskolin-treated cells increased up to 7-fold in a dose dependent manner. Increasing concentrations of forskolin also led to an increase in bAR population, with a maximum increase of approximately 40-60% at 10 uM forskolin. A maximum increase of 40-50% in the quantity of MHC was observed at 0.2 uM forskolin, but higher concentrations of forskolin reduced the quantity of MHC back to control levels. At 0.2 uM forskolin, intracellular levels of cAMP were higher by approximately 35%, and the (beta)AR population was higher by approximately 30%. Neither the number of muscle nuclei fused into myotubes nor the percentage of nuclei in myotubes were affected by forskolin at any of the concentrations studied.

  12. Production of human monoclonal antibody in eggs of chimeric chickens.

    PubMed

    Zhu, Lei; van de Lavoir, Marie-Cecile; Albanese, Jenny; Beenhouwer, David O; Cardarelli, Pina M; Cuison, Severino; Deng, David F; Deshpande, Shrikant; Diamond, Jennifer H; Green, Lynae; Halk, Edward L; Heyer, Babette S; Kay, Robert M; Kerchner, Allyn; Leighton, Philip A; Mather, Christine M; Morrison, Sherie L; Nikolov, Zivko L; Passmore, David B; Pradas-Monne, Alicia; Preston, Benjamin T; Rangan, Vangipuram S; Shi, Mingxia; Srinivasan, Mohan; White, Steven G; Winters-Digiacinto, Peggy; Wong, Susan; Zhou, Wen; Etches, Robert J

    2005-09-01

    The tubular gland of the chicken oviduct is an attractive system for protein expression as large quantities of proteins are deposited in the egg, the production of eggs is easily scalable and good manufacturing practices for therapeutics from eggs have been established. Here we examined the ability of upstream and downstream DNA sequences of ovalbumin, a protein produced exclusively in very high quantities in chicken egg white, to drive tissue-specific expression of human mAb in chicken eggs. To accommodate these large regulatory regions, we established and transfected lines of chicken embryonic stem (cES) cells and formed chimeras that express mAb from cES cell-derived tubular gland cells. Eggs from high-grade chimeras contained up to 3 mg of mAb that possesses enhanced antibody-dependent cellular cytotoxicity (ADCC), nonantigenic glycosylation, acceptable half-life, excellent antigen recognition and good rates of internalization.

  13. Electro-quasistatic analysis of an electrostatic induction micromotor using the cell method.

    PubMed

    Monzón-Verona, José Miguel; Santana-Martín, Francisco Jorge; García-Alonso, Santiago; Montiel-Nelson, Juan Antonio

    2010-01-01

    An electro-quasistatic analysis of an induction micromotor has been realized by using the Cell Method. We employed the direct Finite Formulation (FF) of the electromagnetic laws, hence, avoiding a further discretization. The Cell Method (CM) is used for solving the field equations at the entire domain (2D space) of the micromotor. We have reformulated the field laws in a direct FF and analyzed physical quantities to make explicit the relationship between magnitudes and laws. We applied a primal-dual barycentric discretization of the 2D space. The electric potential has been calculated on each node of the primal mesh using CM. For verification purpose, an analytical electric potential equation is introduced as reference. In frequency domain, results demonstrate the error in calculating potential quantity is neglected (<3‰). In time domain, the potential value in transient state tends to the steady state value.

  14. Electro-Quasistatic Analysis of an Electrostatic Induction Micromotor Using the Cell Method

    PubMed Central

    Monzón-Verona, José Miguel; Santana-Martín, Francisco Jorge; García–Alonso, Santiago; Montiel-Nelson, Juan Antonio

    2010-01-01

    An electro-quasistatic analysis of an induction micromotor has been realized by using the Cell Method. We employed the direct Finite Formulation (FF) of the electromagnetic laws, hence, avoiding a further discretization. The Cell Method (CM) is used for solving the field equations at the entire domain (2D space) of the micromotor. We have reformulated the field laws in a direct FF and analyzed physical quantities to make explicit the relationship between magnitudes and laws. We applied a primal-dual barycentric discretization of the 2D space. The electric potential has been calculated on each node of the primal mesh using CM. For verification purpose, an analytical electric potential equation is introduced as reference. In frequency domain, results demonstrate the error in calculating potential quantity is neglected (<3‰). In time domain, the potential value in transient state tends to the steady state value. PMID:22163397

  15. Impact of the exopolysaccharide layer on biofilms, adhesion and resistance to stress in Lactobacillus johnsonii FI9785.

    PubMed

    Dertli, Enes; Mayer, Melinda J; Narbad, Arjan

    2015-02-04

    The bacterial cell surface is a crucial factor in cell-cell and cell-host interactions. Lactobacillus johnsonii FI9785 produces an exopolysaccharide (EPS) layer whose quantity and composition is altered in mutants that harbour genetic changes in their eps gene clusters. We have assessed the effect of changes in EPS production on cell surface characteristics that may affect the ability of L. johnsonii to colonise the poultry host and exclude pathogens. Analysis of physicochemical cell surface characteristics reflected by Zeta potential and adhesion to hexadecane showed that an increase in EPS gave a less negative, more hydrophilic surface and reduced autoaggregation. Autoaggregation was significantly higher in mutants that have reduced EPS, indicating that EPS can mask surface structures responsible for cell-cell interactions. EPS also affected biofilm formation, but here the quantity of EPS produced was not the only determinant. A reduction in EPS production increased bacterial adhesion to chicken gut explants, but made the bacteria less able to survive some stresses. This study showed that manipulation of EPS production in L. johnsonii FI9785 can affect properties which may improve its performance as a competitive exclusion agent, but that positive changes in adhesion may be compromised by a reduction in the ability to survive stress.

  16. Cell Wall Chemical Composition of Enterococcus faecalis in the Viable but Nonculturable State

    PubMed Central

    Signoretto, Caterina; del Mar Lleò, Maria; Tafi, Maria Carla; Canepari, Pietro

    2000-01-01

    The viable but nonculturable (VBNC) state is a survival mechanism adopted by many bacteria (including those of medical interest) when exposed to adverse environmental conditions. In this state bacteria lose the ability to grow in bacteriological media but maintain viability and pathogenicity and sometimes are able to revert to regular division upon restoration of normal growth conditions. The aim of this work was to analyze the biochemical composition of the cell wall of Enterococcus faecalis in the VBNC state in comparison with exponentially growing and stationary cells. VBNC enterococcal cells appeared as slightly elongated and were endowed with a wall more resistant to mechanical disruption than dividing cells. Analysis of the peptidoglycan chemical composition showed an increase in total cross-linking, which rose from 39% in growing cells to 48% in VBNC cells. This increase was detected in oligomers of a higher order than dimers, such as trimers (24% increase), tetramers (37% increase), pentamers (65% increase), and higher oligomers (95% increase). Changes were also observed in penicillin binding proteins (PBPs), the enzymes involved in the terminal stages of peptidoglycan assembly, with PBPs 5 and 1 being prevalent, and in autolytic enzymes, with a threefold increase in the activity of latent muramidase-1 in E. faecalis in the VBNC state. Accessory wall polymers such as teichoic acid and lipoteichoic acid proved unchanged and doubled in quantity, respectively, in VBNC cells in comparison to dividing cells. It is suggested that all these changes in the cell wall of VBNC enterococci are specific to this particular physiological state. This may provide indirect confirmation of the viability of these cells. PMID:10788366

  17. THE MECHANISM OF THE INHIBITION OF HEMOLYSIS

    PubMed Central

    Ponder, Eric

    1946-01-01

    This paper is concerned with hemolytic systems containing sodium taurocholate, sodium glycocholate, or saponin, and either human red cells or the ghosts (stromata) of human red cells. The lysins are allowed to react with the cells for a short time (1 minute or less), and with the ghosts for a long time (4 hours), and the quantity of lysin remaining in the bulk phase, after the removal of the cells or of the ghosts, is found by (a) colorimetric methods, and (b) methods which measure its hemolytic activity. In the experiments in which the lysins react with the cells for a time so short that none of them is hemolyzed, it is found: (1) that the suspension medium of a cell suspension contains inhibitory substances which render a small amount of the lysin non-lytic, (2) that on the addition of the lysin to the cell suspension, a further and much larger amount of lysin is rendered non-lytic, and (3) that the quantity of chromogenic material in the bulk phase, after the lysin has been in contact with the cells and the latter have been removed, is substantially the same as that initially present. No appreciable quantity of lysin, accordingly, accumulates in increased concentration at the cell surfaces. The results of the colorimetric determinations show that the apparent disappearance of lysin from the bulk phase, once thought to be due to an accumulation of lysin at the cell surfaces, is the result either of an inhibitory process or of a sudden utilization of lysin unrecognized by existing theory. In the experiments in which the lysins react with stromata for 4 hours, it is found: (1) that the suspension medium of a stroma suspension contains inhibitory substances which render some of the lysin non-lytic, (2) that when the lysin reacts with the stromata over a period of time, a much greater quantity of lysin is rendered non-lytic, and (3) that the concentration of chromogenic material in the bulk phase of the lysin-stroma system, after 4 hours of reaction, is substantially the same as it was initially. The observations can be accounted for by supposing that the lysin molecules are as chromogenic after reacting with the cell components as before, and by their not being bound to the cell or to the ghost, but diffusing back, combined with the components with which they have reacted, into the bulk phase. Such a process would have similarities to the process of penetration and breakdown of mixed protein-lipoid films. Because it is not possible at present to decide how much of the lysin is rendered inert because of utilization in a reaction with cell components, and how much because of the effect of inhibitory substances, difficulties and uncertainties arise in connection with the expressions which have been used to describe the kinetics of hemolysis. PMID:19873457

  18. Human interleukin for DA cells or leukemia inhibitory factor is released by Vero cells in human embryo coculture.

    PubMed

    Papaxanthos-Roche, A; Taupin, J L; Mayer, G; Daniel, J Y; Moreau, J F

    1994-09-01

    In the light of the newly discovered implications of human interleukin for DA cells and leukemia inhibitory factor in embryology, we searched for the presence of this soluble cytokine in the supernatant of Vero cell coculture systems. Using a bioassay as well as a specific ELISA, we demonstrated that Vero cells are able to release large quantities of human interleukin for DA cells and leukemia inhibitory factor in the embryo-growing medium of such cocultures.

  19. Self-digitization microfluidic chip for absolute quantification of mRNA in single cells.

    PubMed

    Thompson, Alison M; Gansen, Alexander; Paguirigan, Amy L; Kreutz, Jason E; Radich, Jerald P; Chiu, Daniel T

    2014-12-16

    Quantification of mRNA in single cells provides direct insight into how intercellular heterogeneity plays a role in disease progression and outcomes. Quantitative polymerase chain reaction (qPCR), the current gold standard for evaluating gene expression, is insufficient for providing absolute measurement of single-cell mRNA transcript abundance. Challenges include difficulties in handling small sample volumes and the high variability in measurements. Microfluidic digital PCR provides far better sensitivity for minute quantities of genetic material, but the typical format of this assay does not allow for counting of the absolute number of mRNA transcripts samples taken from single cells. Furthermore, a large fraction of the sample is often lost during sample handling in microfluidic digital PCR. Here, we report the absolute quantification of single-cell mRNA transcripts by digital, one-step reverse transcription PCR in a simple microfluidic array device called the self-digitization (SD) chip. By performing the reverse transcription step in digitized volumes, we find that the assay exhibits a linear signal across a wide range of total RNA concentrations and agrees well with standard curve qPCR. The SD chip is found to digitize a high percentage (86.7%) of the sample for single-cell experiments. Moreover, quantification of transferrin receptor mRNA in single cells agrees well with single-molecule fluorescence in situ hybridization experiments. The SD platform for absolute quantification of single-cell mRNA can be optimized for other genes and may be useful as an independent control method for the validation of mRNA quantification techniques.

  20. Antifungal mechanism of the combination of Cinnamomum verum and Pelargonium graveolens essential oils with fluconazole against pathogenic Candida strains.

    PubMed

    Essid, Rym; Hammami, Majdi; Gharbi, Dorra; Karkouch, Ines; Hamouda, Thouraya Ben; Elkahoui, Salem; Limam, Ferid; Tabbene, Olfa

    2017-09-01

    The present study aimed to investigate the anti-Candida activity of ten essential oils (EOs) and to evaluate their potential synergism with conventional drugs. The effect on secreted aspartic protease (SAP) activity and the mechanism of action were also explored. The antifungal properties of essential oils were investigated using standard micro-broth dilution assay. Only Cinnamomum verum, Thymus capitatus, Syzygium aromaticum, and Pelargonium graveolens exhibited a broad spectrum of activity against a variety of pathogenic Candida strains. Chemical composition of active essential oils was performed by gas chromatography-mass spectrometry (GC-MS). Synergistic effect was observed with the combinations C. verum/fluconazole and P. graveolens/fluconazole, with FIC value 0.37. Investigation of the mechanism of action revealed that C. verum EO reduced the quantity of ergosterol to 83%. A total inhibition was observed for the combination C. verum/fluconazole. However, P. graveolens EO may disturb the permeability barrier of the fungal cell wall. An increase of MIC values of P. graveolens EO and the combination with fluconazole was observed with osmoprotectants (sorbitol and PEG6000). Furthermore, the combination with fluconazole may affect ergosterol biosynthesis and disturb fatty acid homeostasis in C. albicans cells as the quantity of ergosterol and oleic acid was reduced to 52.33 and 72%, respectively. The combination of P. graveolens and C. verum EOs with fluconazole inhibited 78.31 and 64.72% SAP activity, respectively. To our knowledge, this is the first report underlying the mechanism of action and the inhibitory effect of SAP activity of essential oils in synergy with fluconazole. Naturally occurring phytochemicals C. verum and P. graveolens could be effective candidate to enhance the efficacy of fluconazole-based therapy of C. albicans infections.

  1. Plant Responses to Brief Touching: A Mechanism for Early Neighbour Detection?

    PubMed

    Markovic, Dimitrije; Nikolic, Neda; Glinwood, Robert; Seisenbaeva, Gulaim; Ninkovic, Velemir

    2016-01-01

    In natural habitats plants can be exposed to brief and light contact with neighbouring plants. This mechanical stimulus may represent a cue that induces responses to nearby plants. However, little is known about the effect of touching on plant growth and interaction with insect herbivores. To simulate contact between plants, a soft brush was used to apply light and brief mechanical stimuli to terminal leaves of potato Solanum tuberosum L. The number of non-glandular trichomes on the leaf surface was counted on images made by light microscope while glandular trichomes and pavement cells were counted on images made under scanning electronic microscope. Volatile compounds were identified and quantified using coupled gas chromatography-mass spectrometry (GC-MS). Treated plants changed their pattern of biomass distribution; they had lower stem mass fraction and higher branch and leaf mass fraction than untouched plants. Size, weight and number of tubers were not significantly affected. Touching did not cause trichome damage nor change their total number on touched terminal leaves. However, on primary leaves the number of glandular trichomes and pavement cells was significantly increased. Touching altered the volatile emission of treated plants; they released higher quantities of the sesquiterpenes (E)-β-caryophyllene, germacrene D-4-ol and (E)-nerolidol, and lower quantities of the terpenes (E)-ocimene and linalool, indicating a systemic effect of the treatment. The odour of touched plants was significantly less preferred by the aphids Macrosiphum euphorbiae and Myzus persicae compared to odour of untouched plants. The results suggest that light contact may have a potential role in the detection of neighbouring plants and may affect plant-insect interactions.

  2. Plant Responses to Brief Touching: A Mechanism for Early Neighbour Detection?

    PubMed Central

    Markovic, Dimitrije; Nikolic, Neda; Glinwood, Robert; Seisenbaeva, Gulaim; Ninkovic, Velemir

    2016-01-01

    In natural habitats plants can be exposed to brief and light contact with neighbouring plants. This mechanical stimulus may represent a cue that induces responses to nearby plants. However, little is known about the effect of touching on plant growth and interaction with insect herbivores. To simulate contact between plants, a soft brush was used to apply light and brief mechanical stimuli to terminal leaves of potato Solanum tuberosum L. The number of non-glandular trichomes on the leaf surface was counted on images made by light microscope while glandular trichomes and pavement cells were counted on images made under scanning electronic microscope. Volatile compounds were identified and quantified using coupled gas chromatography–mass spectrometry (GC-MS). Treated plants changed their pattern of biomass distribution; they had lower stem mass fraction and higher branch and leaf mass fraction than untouched plants. Size, weight and number of tubers were not significantly affected. Touching did not cause trichome damage nor change their total number on touched terminal leaves. However, on primary leaves the number of glandular trichomes and pavement cells was significantly increased. Touching altered the volatile emission of treated plants; they released higher quantities of the sesquiterpenes (E)-β-caryophyllene, germacrene D-4-ol and (E)-nerolidol, and lower quantities of the terpenes (E)-ocimene and linalool, indicating a systemic effect of the treatment. The odour of touched plants was significantly less preferred by the aphids Macrosiphum euphorbiae and Myzus persicae compared to odour of untouched plants. The results suggest that light contact may have a potential role in the detection of neighbouring plants and may affect plant-insect interactions. PMID:27828995

  3. Nickel-cadmium cells

    NASA Technical Reports Server (NTRS)

    Rubin, E. J.; Turchan, M. J.

    1974-01-01

    A high energy density nickel cadmium cell of aerospace quality was designed. The approach used was to utilize manufacturing techniques which produce highly uniform and controlled starting materials in addition to improvements in the overall design. Parameters controlling the production of plaque and both positive and negative plate were studied. Quantities of these materials were produced and prototype cells were assembled to test the proposed design.

  4. Cardiac mesenchymal progenitors from postmortem cardiac tissues retained cellular characterization.

    PubMed

    Kami, D; Kitani, T; Nakata, M; Gojo, S

    2014-05-01

    Currently, cells for transplantation in regenerative medicine are derived from either autologous or allogeneic tissue. The former has the drawbacks that the quality of donor cells may depend on the condition of the patient, while the quantity of the cells may also be limited. To solve these problems, we investigated the potential of allogeneic cardiac mesenchymal progenitors (CMPs) derived from postmortem hearts, which may be immunologically privileged similar to bone marrow-derived mesenchymal progenitors. We examined whether viable CMPs could be isolated from C57/B6 murine cardiac tissues harvested at 24 hours postmortem. After 2- to 3-week propagation with a high dose of basic fibroblast growth factor, we performed cellular characteristics analyses, which included proliferation and differentiation property flow cytometry and microarray analyses. Postmortem CMPs had a longer lag phase after seeding than CMPs obtained from living tissues, but otherwise had similar characteristics in all the analyses. In addition, global gene expression analysis by microarray showed that cells derived from postmortem and living tissues had similar characteristics. These results indicate that allogeneic postmortem CMPs have potential for cell transplantation because they circumvent the issue of both the quality and quantity of donor cells. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Mechanical contribution of lamellar and interlamellar elastin along the mouse aorta.

    PubMed

    Clark, T E; Lillie, M A; Vogl, A W; Gosline, J M; Shadwick, R E

    2015-10-15

    The mechanical properties of aortic elastin vary regionally, but the microstructural basis for this variation is unknown. This study was designed to identify the relative contributions of lamellar and interlamellar elastin to circumferential load bearing in the mouse thoracic and abdominal aortas. Forces developed in uniaxial tests of samples of fresh and autoclaved aorta were correlated with elastin content and morphology obtained from histology and multiphoton laser scanning microscopy. Autoclaving should render much of the interlamellar elastin mechanically incompetent. In autoclaved tissue force per unit sample width correlated with lamellar elastin content (P≪0.001) but not total elastin content. In fresh tissue at low strain where elastin dominates the mechanical response, forces were higher than in the autoclaved tissue, but force did not correlate with total elastin content. Therefore although interlamellar elastin likely contributed to the stiffness in the fresh aorta, its contribution appeared not in proportion to its quantity. In both fresh and autoclaved tissue, elastin stiffness consistently decreased along the abdominal aorta, a key area for aneurysm development, and this difference could not be fully accounted for on the basis of either lamellar or total elastin content. These findings are relevant to the development of mathematical models of arterial mechanics, particularly for mouse models of arterial diseases involving elastic tissue. In microstructural based models the quantity of each mural constituent determines its contribution to the total response. This study shows elastin's mechanical response cannot necessarily be accounted for on the basis of fibre quantity, orientation, and modulus. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. A proposed performance index for galactic cosmic ray shielding materials

    NASA Technical Reports Server (NTRS)

    Wilson, John W.; Wood, J. S.; Shinn, Judy L.; Cucinotta, Francis A.; Nealy, John E.

    1993-01-01

    In past studies, the reductions in absorbed dose and dose equivalent due to choice of material composition have been used to indicate shield effectiveness against exposure to galactic cosmic rays. However, these quantities are highly inaccurate in assessing shield effectiveness for protection against the biological effects of long-term exposure to the galactic heavy ions. A new quantity for shield performance is defined that correlates well with cell killing and cell transformation behind various shield thicknesses and materials. In addition, a relative performance index is identified that is inversely related to biological injury for different materials at a fixed shield mass and is directly related to the ratio of the fourth- and the second-order linear energy transfer (LET) moments.

  7. Glide-Plane Symmetry and Superconducting Gap Structure of Iron-Based Superconductors

    NASA Astrophysics Data System (ADS)

    Maier, Thomas

    This talk will provide a review of the implications of the glide plane symmetry of a single Fe-pnictide/chalcogen plane on the structure of the superconducting gap. It will be shown that `` η-pairing'' with non-zero total momentum occurs inevitably in this system, but that its contribution to the superconducting condensate has the usual even parity symmetry and time reversal symmetry is preserved. I will demonstrate that for a single plane the gap function, which appears in physical quantities, is identical to that found in 1 Fe per unit cell pseudo-crystal momentum calculations and discuss the effects of the symmetry breaking out-of-plane hopping integrals in three dimensions. A portion of this research was conducted at the Center for Nanophase Materials Sciences, which is a DOE Office of Science User Facility.

  8. Anionic polymer, poly(methyl vinyl ether-maleic anhydride)-coated beads-based capture of human influenza A and B virus.

    PubMed

    Sakudo, Akikazu; Baba, Koichi; Tsukamoto, Megumi; Sugimoto, Atsuko; Okada, Takashi; Kobayashi, Takanori; Kawashita, Norihito; Takagi, Tatsuya; Ikuta, Kazuyoshi

    2009-01-15

    An anionic magnetic beads-based method was developed for the capture of human influenza A and B viruses from nasal aspirates, allantoic fluid and culture medium. A polymer, poly(methyl vinyl ether-maleic anhydride) [poly(MVE-MA)], was used to endow magnetic beads with a negative charge and bioadhesive properties. After incubation with samples containing human influenza virus, the beads were separated from supernatants by applying a magnetic field. The adsorption [corrected] of the virus by the beads was confirmed by hemagglutinin assay, immunochromatography, Western blotting, egg infection, and cell infection. Successful capture was proved using 5 H1N1 influenza A viruses, 10 H3N2 influenza A viruses, and 6 influenza B viruses. Furthermore, the infectivity in chicken embryonated eggs and Madin-Darby canine kidney (MDCK) cells of the captured human influenza virus was similar to that of the total viral quantity of starting materials. Therefore, this method of capture using magnetic beads coated with poly(MVE-MA) can be broadly used for the recovery of infectious human influenza viruses.

  9. Microarrays for Undergraduate Classes

    ERIC Educational Resources Information Center

    Hancock, Dale; Nguyen, Lisa L.; Denyer, Gareth S.; Johnston, Jill M.

    2006-01-01

    A microarray experiment is presented that, in six laboratory sessions, takes undergraduate students from the tissue sample right through to data analysis. The model chosen, the murine erythroleukemia cell line, can be easily cultured in sufficient quantities for class use. Large changes in gene expression can be induced in these cells by…

  10. Phosphorus retention by fly-ash amended filter media in aged bioretention cells

    USDA-ARS?s Scientific Manuscript database

    Bioretention cells (BRCs) have shown potential for stormwater quantity and quality control. However, the phosphorus (P) removal in BRC has been variable due to differences of soil properties in filter media. The objectives of this research were to identify and evaluate P accumulation in filter media...

  11. Collective synthesis of natural products by means of organocascade catalysis.

    PubMed

    Jones, Spencer B; Simmons, Bryon; Mastracchio, Anthony; MacMillan, David W C

    2011-07-13

    Organic chemists are now able to synthesize small quantities of almost any known natural product, given sufficient time, resources and effort. However, translation of the academic successes in total synthesis to the large-scale construction of complex natural products and the development of large collections of biologically relevant molecules present significant challenges to synthetic chemists. Here we show that the application of two nature-inspired techniques, namely organocascade catalysis and collective natural product synthesis, can facilitate the preparation of useful quantities of a range of structurally diverse natural products from a common molecular scaffold. The power of this concept has been demonstrated through the expedient, asymmetric total syntheses of six well-known alkaloid natural products: strychnine, aspidospermidine, vincadifformine, akuammicine, kopsanone and kopsinine. ©2011 Macmillan Publishers Limited. All rights reserved

  12. Using genetic algorithm to determine the optimal order quantities for multi-item multi-period under warehouse capacity constraints in kitchenware manufacturing

    NASA Astrophysics Data System (ADS)

    Saraswati, D.; Sari, D. K.; Johan, V.

    2017-11-01

    The study was conducted on a manufacturer that produced various kinds of kitchenware with kitchen sink as the main product. There were four types of steel sheets selected as the raw materials of the kitchen sink. The problem was the manufacturer wanted to determine how much steel sheets to order from a single supplier to meet the production requirements in a way to minimize the total inventory cost. In this case, the economic order quantity (EOQ) model was developed using all-unit discount as the price of steel sheets and the warehouse capacity was limited. Genetic algorithm (GA) was used to find the minimum of the total inventory cost as a sum of purchasing cost, ordering cost, holding cost and penalty cost.

  13. In vitro and in vivo infectivity and pathogenicity of the lymphoid cell-derived woodchuck hepatitis virus.

    PubMed

    Lew, Y Y; Michalak, T I

    2001-02-01

    Woodchuck hepatitis virus (WHV) and human hepatitis B virus are closely related, highly hepatotropic mammalian DNA viruses that also replicate in the lymphatic system. The infectivity and pathogenicity of hepadnaviruses propagating in lymphoid cells are under debate. In this study, hepato- and lymphotropism of WHV produced by naturally infected lymphoid cells was examined in specifically established woodchuck hepatocyte and lymphoid cell cultures and coculture systems, and virus pathogenicity was tested in susceptible animals. Applying PCR-based assays discriminating between the total pool of WHV genomes and covalently closed circular DNA (cccDNA), combined with enzymatic elimination of extracellular viral sequences potentially associated with the cell surface, our study documents that virus replicating in woodchuck lymphoid cells is infectious to homologous hepatocytes and lymphoid cells in vitro. The productive replication of WHV from lymphoid cells in cultured hepatocytes was evidenced by the appearance of virus-specific DNA, cccDNA, and antigens, transmissibility of the virus through multiple passages in hepatocyte cultures, and the ability of the passaged virus to infect virus-naive animals. The data also revealed that WHV from lymphoid cells can initiate classical acute viral hepatitis in susceptible animals, albeit small quantities (approximately 10(3) virions) caused immunovirologically undetectable (occult) WHV infection that engaged the lymphatic system but not the liver. Our results provide direct in vitro and in vivo evidence that lymphoid cells in the infected host support propagation of infectious hepadnavirus that has the potential to induce hepatitis. They also emphasize a principal role of the lymphatic system in the maintenance and dissemination of hepadnavirus infection, particularly when infection is induced by low virus doses.

  14. Gigantism in a Bacterium, Epulopiscium fishelsoni, Correlates with Complex Patterns in Arrangement, Quantity, and Segregation of DNA

    PubMed Central

    Bresler, V.; Montgomery, W. L.; Fishelson, L.; Pollak, P. E.

    1998-01-01

    Epulopiscium fishelsoni, gut symbiont of the brown surgeonfish (Acanthurus nigrofuscus) in the Red Sea, attains a larger size than any other eubacterium, varies 10- to 20-fold in length (and >2,000-fold in volume), and undergoes a complex daily life cycle. In early morning, nucleoids contain highly condensed DNA in elongate, chromosome-like structures which are physically separated from the general cytoplasm. Cell division involves production of two (rarely three) nucleoids within a cell, deposition of cell walls around expanded nucleoids, and emergence of daughter cells from the parent cell. Fluorescence measurements of DNA, RNA, and other cell components indicate the following. DNA quantity is proportional to cell volume over cell lengths of ∼30 μm to >500 μm. For cells of a given size, nucleoids of cells with two nucleoids (binucleoid) contain approximately equal amounts of DNA. And each nucleoid of a binucleoid cell contains one-half the DNA of the single nucleoid in a uninucleoid cell of the same size. The life cycle involves approximately equal subdivision of DNA among daughter cells, formation of apical caps of condensed DNA from previously decondensed and diffusely distributed DNA, and “pinching” of DNA near the middle of the cell in the absence of new wall formation. Mechanisms underlying these patterns remain unclear, but formation of daughter nucleoids and cells occurs both during diurnal periods of host feeding and bacterial cell growth and during nocturnal periods of host inactivity when mean bacterial cell size declines. PMID:9791108

  15. Relative quantity judgments between discrete spatial arrays by chimpanzees (Pan troglodytes) and New Zealand robins (Petroica longipes).

    PubMed

    Garland, Alexis; Beran, Michael J; McIntyre, Joseph; Low, Jason

    2014-08-01

    Quantity discrimination for items spread across spatial arrays was investigated in chimpanzees (Pan troglodytes) and North Island New Zealand robins (Petroica longipes), with the aim of examining the role of spatial separation on the ability of these 2 species to sum and compare nonvisible quantities which are both temporally and spatially separated, and to assess the likely mechanism supporting such summation performance. Birds and chimpanzees compared 2 sets of discrete quantities of items that differed in number. Six quantity comparisons were presented to both species: 1v2, 1v3, 1v5, 2v3, 2v4, and 2v5. Each was distributed 1 at a time across 2 7-location arrays. Every individual item was viewed 1 at a time and hidden, with no more than a single item in each location of an array, in contrast to a format where all items were placed together into 2 single locations. Subjects responded by selecting 1 of the 2 arrays and received the entire quantity of food items hidden within that array. Both species performed better than chance levels. The ratio of items between sets was a significant predictor of performance in the chimpanzees, but it was not significant for robins. Instead, the absolute value of the smaller quantity of items presented was the significant factor in robin responses. These results suggest a species difference for this task when considering various dimensions such as ratio or total number of items in quantity comparisons distributed across discrete 7-location arrays.

  16. Structural analysis of cell wall polysaccharides using PACE

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mortimer, Jennifer C.

    The plant cell wall is composed of many complex polysaccharides. The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. Many techniques to characterize polysaccharide structure are complicated, requiring expensive equipment and specialized operators e.g. NMR, MALDI-MS. PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al. 2002). Whilst the method here describes xylan analysis, it can be applied (by use of the appropriate glycosyl hydrolase) to any cell wall polysaccharide.

  17. The Cholinergic Synapse International Symposium Held in Berlin, Germany on 23-27 September 1990

    DTIC Science & Technology

    1990-09-27

    DNA into Spodoptera frugiperda cells and recombinant virus carrying the AChE gene were iden- tified by filter-hybridisation and isolated. Purified...enzyme when expressed in COS cells. Expression in the Spodoptera -baculovirus system allows for production of large quantities of AChE and its mutant...recombinant- virus clones were multiplied and used for infection of S. frugiperda cells. Infected cells were harvested 2.4 days following infection and

  18. Imaging of blood antigen distribution on blood cells by thermal lens microscopy

    NASA Astrophysics Data System (ADS)

    Kimura, Hiroko; Sekiguchi, Kazuya; Nagao, Fumiko; Mukaida, Masahiro; Kitamori, Takehiko; Sawada, Tsuguo

    2000-05-01

    Blood group antigens on a cell were measured by a new microscopic method, i.e. thermal lens microscopy which involves spectrometry using a laser-induced thermal-lens effect. The blood group antigen was immunologically stained using antibody labeled with colloidal gold. Human leukocyte antigens (HLA) on lymphocytes and mononuclear leukocytes were observed by the thermal lens microscope, and Lewis blood group antigens on erythrocytes and polymorphonuclear leukocytes were also observed. The antigen distribution on each cell-surface was imaged using this technique. In spite of convex surface of living cells, colloidal gold was correctly quantified by adjusting the deviation of the focal point of the probe laser by the phase of the signal. In the measurement of leukocyte antigens, antigens of HLA-A, -B, -C loci on the lymphocytes were identified and quantitated by using a single cell. The image of HLA-A, -B, -C antigen distribution on a mononuclear leukocyte was obtained. In the measurement of erythrocyte antigens, a small quantity of Lewis antigens was detected on the cord erythrocytes. Localized small quantities of membrane antigens are better quantitated without extraction or cytolysis. Our thermal lens microscope is a powerful and highly sensitive analytical tool for detecting and quantitating localized antigens in single cells and/or cell-surface-associated molecules.

  19. Glucose Starvation Alters Heat Shock Response, Leading to Death of Wild Type Cells and Survival of MAP Kinase Signaling Mutant

    PubMed Central

    Higgins, LeeAnn; Markowski, Todd; Brambl, Robert

    2016-01-01

    A moderate heat shock induces Neurospora crassa to synthesize large quantities of heat shock proteins that are protective against higher, otherwise lethal temperatures. However, wild type cells do not survive when carbohydrate deprivation is added to heat shock. In contrast, a mutant strain defective in a stress-activated protein kinase does survive the combined stresses. In order to understand the basis for this difference in survival, we have determined the relative levels of detected proteins in the mutant and wild type strain during dual stress, and we have identified gene transcripts in both strains whose quantities change in response to heat shock or dual stress. These data and supportive experimental evidence point to reasons for survival of the mutant strain. By using alternative respiratory mechanisms, these cells experience less of the oxidative stress that proves damaging to wild type cells. Of central importance, mutant cells recycle limited resources during dual stress by undergoing autophagy, a process that we find utilized by both wild type and mutant cells during heat shock. Evidence points to inappropriate activation of TORC1, the central metabolic regulator, in wild type cells during dual stress, based upon behavior of an additional signaling mutant and inhibitor studies. PMID:27870869

  20. Endophytic fungi specifically introduce novel metabolites into grape flesh cells in vitro

    PubMed Central

    Ao, Xiu-Jin; Ren, An-Yun; Zhang, Han-Bo

    2018-01-01

    Since endophytes can affect metabolism of host plants, they are expected to be used to improve crop quality, especially for crops with organoleptic sensitive products such as wine grape. However, details of metabolic interactions between endophytes and host plants were less understood. In this work, we used high pressure liquid chromatography (HPLC) to analyze the metabolites of fruit flesh cells of grape treated with dual culture of different endophytic fungal strains (EFS). We observed that the dual-culture with different fungal strains show different metabolites composition in grape cells. In response to different EFS, quantities of detected metabolites in grape cells varied from 6 to 17 in this assay, and 1 to 11 novel metabolites were introduced into metabolome of grape cells. Dual-culture with fungal strains CS2, RH16 and RH5 introduced the highest quantities (10 or 11) of novel metabolites in grape cells. More importantly, the modification of metabolic profiles in grape cells via fungal endophytes appeared to be fungal strain/genus-specificity. Overall, this work revealed that introduction of specific metabolites in host plants may be one consequence during the process of endophytes-host metabolic interactions, which raise the possibility to shape grape qualities and characteristics using tool of fungal endophytes. PMID:29734364

  1. Rapid high-throughput cloning and stable expression of antibodies in HEK293 cells.

    PubMed

    Spidel, Jared L; Vaessen, Benjamin; Chan, Yin Yin; Grasso, Luigi; Kline, J Bradford

    2016-12-01

    Single-cell based amplification of immunoglobulin variable regions is a rapid and powerful technique for cloning antigen-specific monoclonal antibodies (mAbs) for purposes ranging from general laboratory reagents to therapeutic drugs. From the initial screening process involving small quantities of hundreds or thousands of mAbs through in vitro characterization and subsequent in vivo experiments requiring large quantities of only a few, having a robust system for generating mAbs from cloning through stable cell line generation is essential. A protocol was developed to decrease the time, cost, and effort required by traditional cloning and expression methods by eliminating bottlenecks in these processes. Removing the clonal selection steps from the cloning process using a highly efficient ligation-independent protocol and from the stable cell line process by utilizing bicistronic plasmids to generate stable semi-clonal cell pools facilitated an increased throughput of the entire process from plasmid assembly through transient transfections and selection of stable semi-clonal cell pools. Furthermore, the time required by a single individual to clone, express, and select stable cell pools in a high-throughput format was reduced from 4 to 6months to only 4 to 6weeks. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  2. 30 CFR 15.21 - Tolerances for ingredients.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...; (c) Carbonaceous materials: ±3 percent; and (d) Moisture and ingredients other than specified in... Moisture and Other Ingredients Quantity of ingredients (as percent of total explosive or sheath) Tolerance...

  3. 30 CFR 15.21 - Tolerances for ingredients.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...; (c) Carbonaceous materials: ±3 percent; and (d) Moisture and ingredients other than specified in... Moisture and Other Ingredients Quantity of ingredients (as percent of total explosive or sheath) Tolerance...

  4. 30 CFR 15.21 - Tolerances for ingredients.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...; (c) Carbonaceous materials: ±3 percent; and (d) Moisture and ingredients other than specified in... Moisture and Other Ingredients Quantity of ingredients (as percent of total explosive or sheath) Tolerance...

  5. 30 CFR 15.21 - Tolerances for ingredients.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...; (c) Carbonaceous materials: ±3 percent; and (d) Moisture and ingredients other than specified in... Moisture and Other Ingredients Quantity of ingredients (as percent of total explosive or sheath) Tolerance...

  6. Odor intensity and characterization of jet exhaust and chemical analytical measurements

    NASA Technical Reports Server (NTRS)

    Kendall, D. A.; Levins, P. L.

    1973-01-01

    Odor and chemical analyses were carried out on the exhaust samples from a J-57 combustor can operated over a range of inlet conditions, and with several fuel types and nozzle modifications. The odor characteristics and total intensity of odor for each exhaust were determined over a range of dilutions to allow for a least squares determination of the intensity at 1,000 to 1 dilutions. Analytical measures included the concentration of total hydrocarbons and the concentrations of aromatic organic species and oxygenated organic species from collected samples which were taken concurrently. A correlation was found between the concentration of the odorous oxygenated fraction and the total intensity of aroma. Inlet operating conditions and nozzle modifications which increase the efficiency of combustion as measured by exhaust gas analyses reduce the odor intensity and the quantity of oxygenates in the exhaust. The type of fuel burned altered the intensity of odor in relation to the quantity of oxygenates produced and, in some instances, changed the odor character.

  7. Quality and Quantity of Sorghum Hydroponic Fodder from Different Varieties and Harvest Time

    NASA Astrophysics Data System (ADS)

    Chrisdiana, R.

    2018-02-01

    This experiment was designed to compare different varieties and harvest time of sorghum hydroponic fodder based on nutrient content and biomass production. Experimental design for fodder productivity was completely randomized design with 2 x 3 factorial, i.e., sorghum varieties (KD 4 and Super-1) and time of harvesting the sorghum hydroponic fodder (8,12 and 16 d). Total biomass and DM production, were affected significantly (p<0.05) on harvest time. Total biomass and nutrient content were increased in longer harvest time. The nutrient content were increased with decreasing total value of DM. Super-1 varieties produce larger biomass and nutrient content higher than KD4 (p<0.05). Based on sorghum hidroponic fodder quality and quantity, sorghum hidroponic fodder with Super-1 varieties harvested at 12 d had a good quality of fodder and it can be alternative of technology providing quality forage and land saving with a short time planting period and continous production.

  8. All-In-One: Advanced preparation of Human Parenchymal and Non-Parenchymal Liver Cells.

    PubMed

    Werner, Melanie; Driftmann, Sabrina; Kleinehr, Kathrin; Kaiser, Gernot M; Mathé, Zotlan; Treckmann, Juergen-Walter; Paul, Andreas; Skibbe, Kathrin; Timm, Joerg; Canbay, Ali; Gerken, Guido; Schlaak, Joerg F; Broering, Ruth

    2015-01-01

    Liver cells are key players in innate immunity. Thus, studying primary isolated liver cells is necessary for determining their role in liver physiology and pathophysiology. In particular, the quantity and quality of isolated cells are crucial to their function. Our aim was to isolate a large quantity of high-quality human parenchymal and non-parenchymal cells from a single liver specimen. Hepatocytes, Kupffer cells, liver sinusoidal endothelial cells, and stellate cells were isolated from liver tissues by collagenase perfusion in combination with low-speed centrifugation, density gradient centrifugation, and magnetic-activated cell sorting. The purity and functionality of cultured cell populations were controlled by determining their morphology, discriminative cell marker expression, and functional activity. Cell preparation yielded the following cell counts per gram of liver tissue: 2.0 ± 0.4 × 10(7) hepatocytes, 1.8 ± 0.5 × 10(6 )Kupffer cells, 4.3 ± 1.9 × 10(5) liver sinusoidal endothelial cells, and 3.2 ± 0.5 × 10(5) stellate cells. Hepatocytes were identified by albumin (95.5 ± 1.7%) and exhibited time-dependent activity of cytochrome P450 enzymes. Kupffer cells expressed CD68 (94.5 ± 1.2%) and exhibited phagocytic activity, as determined with 1 μm latex beads. Endothelial cells were CD146(+) (97.8 ± 1.1%) and exhibited efficient uptake of acetylated low-density lipoprotein. Hepatic stellate cells were identified by the expression of α-smooth muscle actin (97.1 ± 1.5%). These cells further exhibited retinol (vitamin A)-mediated autofluorescence. Our isolation procedure for primary parenchymal and non-parenchymal liver cells resulted in cell populations of high purity and quality, with retained physiological functionality in vitro. Thus, this system may provide a valuable tool for determining liver function and disease.

  9. Tetramer-organizing polyproline-rich peptides differ in CHO cell-expressed and plasma-derived human butyrylcholinesterase tetramers.

    PubMed

    Schopfer, Lawrence M; Lockridge, Oksana

    2016-06-01

    Tetrameric butyrylcholinesterase (BChE) in human plasma is the product of multiple genes, namely one BCHE gene on chromosome 3q26.1 and multiple genes that encode polyproline-rich peptides. The function of the polyproline-rich peptides is to assemble BChE into tetramers. CHO cells transfected with human BChE cDNA express BChE monomers and dimers, but only low quantities of tetramers. Our goal was to identify the polyproline-rich peptides in CHO-cell derived human BChE tetramers. CHO cell-produced human BChE tetramers were purified from serum-free culture medium. Peptides embedded in the tetramerization domain were released from BChE tetramers by boiling and identified by liquid chromatography-tandem mass spectrometry. A total of 270 proline-rich peptides were sequenced, ranging in size from 6-41 residues. The peptides originated from 60 different proteins that reside in multiple cell compartments including the nucleus, cytoplasm, and endoplasmic reticulum. No single protein was the source of the polyproline-rich peptides in CHO cell-expressed human BChE tetramers. In contrast, 70% of the tetramer-organizing peptides in plasma-derived BChE tetramers originate from lamellipodin. No protein source was identified for polyproline peptides containing up to 41 consecutive proline residues. In conclusion, the use of polyproline-rich peptides as a tetramerization motif is documented only for the cholinesterases, but is expected to serve other tetrameric proteins as well. The CHO cell data suggest that the BChE tetramer-organizing peptide can arise from a variety of proteins. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. On total noncommutativity in quantum mechanics

    NASA Astrophysics Data System (ADS)

    Lahti, Pekka J.; Ylinen, Kari

    1987-11-01

    It is shown within the Hilbert space formulation of quantum mechanics that the total noncommutativity of any two physical quantities is necessary for their satisfying the uncertainty relation or for their being complementary. The importance of these results is illustrated with the canonically conjugate position and momentum of a free particle and of a particle closed in a box.

  11. 7 CFR 1124.73 - Payments to producers and to cooperative associations.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... deduct any payment made pursuant to paragraph (c)(1) of this section; and (3) For the total quantity of... dates such milk was received from that producer; (3) The total pounds of butterfat, protein, and other... follows: (1) Partial payment. For each producer who has not discontinued shipments as of the 18th day of...

  12. 7 CFR 1124.73 - Payments to producers and to cooperative associations.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... deduct any payment made pursuant to paragraph (c)(1) of this section; and (3) For the total quantity of... dates such milk was received from that producer; (3) The total pounds of butterfat, protein, and other... follows: (1) Partial payment. For each producer who has not discontinued shipments as of the 18th day of...

  13. 7 CFR 1124.73 - Payments to producers and to cooperative associations.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... deduct any payment made pursuant to paragraph (c)(1) of this section; and (3) For the total quantity of... dates such milk was received from that producer; (3) The total pounds of butterfat, protein, and other... follows: (1) Partial payment. For each producer who has not discontinued shipments as of the 18th day of...

  14. 7 CFR 1124.73 - Payments to producers and to cooperative associations.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... deduct any payment made pursuant to paragraph (c)(1) of this section; and (3) For the total quantity of... dates such milk was received from that producer; (3) The total pounds of butterfat, protein, and other... follows: (1) Partial payment. For each producer who has not discontinued shipments as of the 18th day of...

  15. 7 CFR 1124.73 - Payments to producers and to cooperative associations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... deduct any payment made pursuant to paragraph (c)(1) of this section; and (3) For the total quantity of... dates such milk was received from that producer; (3) The total pounds of butterfat, protein, and other... follows: (1) Partial payment. For each producer who has not discontinued shipments as of the 18th day of...

  16. Trypanosoma congolense: B-lymphocyte responses differ between trypanotolerant and trypanosusceptible cattle.

    PubMed

    Taylor, K A; Lutje, V; Kennedy, D; Authié, E; Boulangé, A; Logan-Henfrey, L; Gichuki, B; Gettinby, G

    1996-06-01

    Trypanosomiasis is a serious constraint to livestock production in sub-Saharan Africa. Some breeds of cattle are genetically more resistant to the pathogenic effects of trypanosome infection. We measured B-cell activation and the quantity and isotype of antibody produced at the cellular level in six trypanotolerant N'Dama and five trypanosusceptible Boran cattle. The frequencies of spleen cells secreting total and parasite-specific IgM and IgG were measured prior to and 16, 28, and 35 days after a primary challenge with Trypanosoma congolense. Boran cattle had higher frequencies of splenic cells secreting IgM specific for trypanosome-derived variable surface glycoprotein (VSG), cysteine protease (congopain, CP), and heat shock protein (hsp 70/BiP) and the nonparasite antigen, ovalbumin, than did N'Dama cattle. In contrast, the number of VSG-specific IgG-secreting cells was significantly greater in N'Dama than in Boran cattle. During infection, low titers of anti-VSG IgM were detected transiently in the serum of all animals. However, N'Dama had significantly more VSG-specific IgG in blood than Boran during infection. The peripheral blood mononuclear cell population of N'Dama cattle contained a higher percentage of surface IgM-positive B-cells prior to and throughout infection than were found in the blood of Boran. In addition, during infection N'Dama cattle had more circulating lymphocytes that could be activated in vitro to undergo differentiation into IgM- and IgG-secreting cells. These findings demonstrate differences in the frequency of trypanosome-specific antibody-secreting cells in the spleen and in the activation state of B-cells in the blood between N'Dama and Boran cattle during a primary infection with T. congolense.

  17. Spiking and Excitatory/Inhibitory Input Dynamics of Barrel Cells in Response to Whisker Deflections of Varying Velocity and Angular Direction.

    PubMed

    Patel, Mainak

    2018-01-15

    The spiking of barrel regular-spiking (RS) cells is tuned for both whisker deflection direction and velocity. Velocity tuning arises due to thalamocortical (TC) synchrony (but not spike quantity) varying with deflection velocity, coupled with feedforward inhibition, while direction selectivity is not fully understood, though may be due partly to direction tuning of TC spiking. Data show that as deflection direction deviates from the preferred direction of an RS cell, excitatory input to the RS cell diminishes minimally, but temporally shifts to coincide with the time-lagged inhibitory input. This work constructs a realistic large-scale model of a barrel; model RS cells exhibit velocity and direction selectivity due to TC input dynamics, with the experimentally observed sharpening of direction tuning with decreasing velocity. The model puts forth the novel proposal that RS→RS synapses can naturally and simply account for the unexplained direction dependence of RS cell inputs - as deflection direction deviates from the preferred direction of an RS cell, and TC input declines, RS→RS synaptic transmission buffers the decline in total excitatory input and causes a shift in timing of the excitatory input peak from the peak in TC input to the delayed peak in RS input. The model also provides several experimentally testable predictions on the velocity dependence of RS cell inputs. This model is the first, to my knowledge, to study the interaction of direction and velocity and propose physiological mechanisms for the stimulus dependence in the timing and amplitude of RS cell inputs. Copyright © 2017 IBRO. Published by Elsevier Ltd. All rights reserved.

  18. [Effect of CsA bleomycin-induced interstitial pulmonary disease in mice].

    PubMed

    Ren, Ying; Yang, Hui; Zhu, Ping; Fan, Chun-mei; Wang, Yan-hong; Li, Jia; Liu, Hui

    2012-03-01

    To observe the therapeutic effect of cyclosporine A (CsA) on bleomycin (BLM) induced pulmonary fibrosis and to investigate its mechanism. One hundred and twenty C57BL/6 female mice were divided randomly into five groups: BLM model group, control saline group, CsA30 mg treatment group, CsA50 mg treatment group and control treatment group. Treatment groups and model groups were administrated BLM intratracheally to induce interstitial pulmonary disease model, with control saline group administrated with equal volume of normal saline instead. Mice in treatment groups were intraperitoneal injected with CsA, while control treatment group were injected with equal volume of normal saline instead. On the 4th, 7th and 14th day after administration, 8 mice of each group were sacrificed, and the peripheral blood was obtained to count total leucocytes with counting chamber and quantify CD4(+); T cells, CD14(+); monocytes and CD19(+); B cells by flow cytometry (FCM). Bronchoalveolar levage fluid was harvested for cell counting and Giemsa staining. Lung tissues were harvested for immunohistochemical staining and pathological examination. The quantity of total leucocyte was higher in BLM model group than those in control saline group.The proportion of CD14(+); T cells and CD19(+);B cells in BLM model group were increased markedly than those in control saline group on the 4th, 7th and 14th day post BLM. With CsA treatment, The proportion of CD14(+); T cells was lower than BLM model group at the same time point, especially on the 4th day. The proportion of CD19(+); B cells were significantly lower than those of BLM model group at the same time point(7 d, 14 d). The total and classification of cells of BLM model group were increased markedly than those in control saline group, and decreased obviously in the treatment groups at the same time point. Examination of lung tissues: With the prolonged time of BLM administration, it showed wider alveolar septum, more collagen deposition, as well as more infiltrating inflammatory cells which consisted of generous lymphocyte and few mononuclear macrophages than those in saline control group. With the prolonged time of CsA injection, the interstitial pulmonary inflammation was remissive, and there was less fibroblast infiltration and collagen deposition in pulmonary interstitium and periphery of bronchiole. Alveolar epithelial cells, bronchiolar epithelial cells, mononuclear macrophages, neutrophils and lymphocytes were demonstrated to express CD147, there was higher CD147 expression in BLM model group than those in CsA treatment groups. CsA may heal BLM induced interstitial pulmonary disease by blocking CD147-CypA interaction, then decreasing chemotaxis for the immunocyte, and reducing migration of immunocytes to the lung and collagen deposition in the lung.

  19. Modules for estimating solid waste from fossil-fuel technologies

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Crowther, M.A.; Thode, H.C. Jr.; Morris, S.C.

    1980-10-01

    Solid waste has become a subject of increasing concern to energy industries for several reasons. Increasingly stringent air and water pollution regulations result in a larger fraction of residuals in the form of solid wastes. Control technologies, particularly flue gas desulfurization, can multiply the amount of waste. With the renewed emphasis on coal utilization and the likelihood of oil shale development, increased amounts of solid waste will be produced. In the past, solid waste residuals used for environmental assessment have tended only to include total quantities generated. To look at environmental impacts, however, data on the composition of the solidmore » wastes are required. Computer modules for calculating the quantities and composition of solid waste from major fossil fuel technologies were therefore developed and are described in this report. Six modules have been produced covering physical coal cleaning, conventional coal combustion with flue gas desulfurization, atmospheric fluidized-bed combustion, coal gasification using the Lurgi process, coal liquefaction using the SRC-II process, and oil shale retorting. Total quantities of each solid waste stream are computed together with the major components and a number of trace elements and radionuclides.« less

  20. Set size and culture influence children's attention to number.

    PubMed

    Cantrell, Lisa; Kuwabara, Megumi; Smith, Linda B

    2015-03-01

    Much research evidences a system in adults and young children for approximately representing quantity. Here we provide evidence that the bias to attend to discrete quantity versus other dimensions may be mediated by set size and culture. Preschool-age English-speaking children in the United States and Japanese-speaking children in Japan were tested in a match-to-sample task where number was pitted against cumulative surface area in both large and small numerical set comparisons. Results showed that children from both cultures were biased to attend to the number of items for small sets. Large set responses also showed a general attention to number when ratio difficulty was easy. However, relative to the responses for small sets, attention to number decreased for both groups; moreover, both U.S. and Japanese children showed a significant bias to attend to total amount for difficult numerical ratio distances, although Japanese children shifted attention to total area at relatively smaller set sizes than U.S. children. These results add to our growing understanding of how quantity is represented and how such representation is influenced by context--both cultural and perceptual. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Quantification and classification of ship scraping waste at Alang-Sosiya, India.

    PubMed

    Srinivasa Reddy, M; Basha, Shaik; Sravan Kumar, V G; Joshi, H V; Ghosh, P K

    2003-12-01

    Alang-Sosiya located on the Western Coast of Gulf of Cambay, is the largest ship recycling yard in the world. Every year on average 365 ships having a mean weight (2.10x10(6)+/-7.82x10(5) LDT) are scrapped. This industry generates a huge quantity of solid waste in the form of broken wood, rubber, insulation materials, paper, metals, glass and ceramics, plastics, leather, textiles, food waste, chemicals, paints, thermocol, sponge, ash, oil mixed sponges, miscellaneous combustible and non-combustible. The quantity and composition of solid waste was collected for a period of three months and the average values are presented in this work. Sosiya had the most waste 15.63 kg/m(2) compared to Alang 10.19 kg/m(2). The combustible solid waste quantity was around 83.0% of the total solid waste available at the yard, which represents an average weight of 9.807 kg/m(2); whereas, non-combustible waste is 1.933 kg/m(2). There is not much difference between the average of total solid waste calculated from the sampling data (96.71 MT/day) and the data provided by the port authorities (96.8 MT/day).

  2. Integrating nutrient bioavailability and co-production links when identifying sustainable diets: How low should we reduce meat consumption?

    PubMed Central

    Gazan, Rozenn; Vieux, Florent; Micard, Valérie; Amiot, Marie-Josèphe; Darmon, Nicole

    2018-01-01

    Background Reducing the consumption of meat and other animal-based products is widely advocated to improve the sustainability of diets in high-income countries. However, such reduction may impair nutritional adequacy, since the bioavailability of key nutrients is higher when they come from animal- vs plant-based foods. Meat reduction may also affect the balance between foods co-produced within the same animal production system. Objective The objective was to assess the impact of introducing nutrient bioavailability and co-production links considerations on the dietary changes needed − especially regarding meat ‒ to improve diet sustainability. Methods Diet optimization with linear and non-linear programming was used to design, for each gender, three modeled diets departing the least from the mean observed French diet (OBS) while reducing by at least 30% the diet-related environmental impacts (greenhouse gas emissions, eutrophication, acidification): i) in the nutrition-environment (NE) model, the fulfillment of recommended dietary allowances for all nutrients was imposed; ii) in the NE-bioavailability (NEB) model, nutritional adequacy was further ensured by accounting for iron, zinc, protein and provitamin A bioavailability; iii) in the NEB-co-production (NEB-CP) model, two links between co-produced animal foods (milk–beef and blood sausage–pork) were additionally included into the models by proportionally co-constraining their respective quantities. The price and environmental impacts of individual foods were assumed to be constant. Results ‘Fruit and vegetables’ and ‘Starches’ quantities increased in all modeled diets compared to OBS. In parallel, total meat and ruminant meat quantities decreased. Starting from 110g/d women’s OBS diet (168g/d for men), total meat quantity decreased by 78%, 67% and 32% for women (68%, 66% and 62% for men) in NE, NEB and NEB-CP diets, respectively. Starting from 36g/d women’s OBS diet (54g/d for men), ruminant meat quantity dropped severely by 84% and 87% in NE and NEB diets for women (80% and 78% for men), whereas it only decreased by 27% in NEB-CP diets (38% for men). The share of energy and proteins of animal origin was similar for the 3 modeled diets (approximately 1/5 of total energy, and 1/2 of protein) and lower than in OBS diet (approximately 1/3 of total energy, and 2/3 of protein). Conclusions Decreasing meat content was strictly needed to achieve more sustainable diets for French adults, but the reduction was less severe when nutrient bioavailability and co-production links were taken into account. PMID:29444098

  3. Integrating nutrient bioavailability and co-production links when identifying sustainable diets: How low should we reduce meat consumption?

    PubMed

    Barré, Tangui; Perignon, Marlène; Gazan, Rozenn; Vieux, Florent; Micard, Valérie; Amiot, Marie-Josèphe; Darmon, Nicole

    2018-01-01

    Reducing the consumption of meat and other animal-based products is widely advocated to improve the sustainability of diets in high-income countries. However, such reduction may impair nutritional adequacy, since the bioavailability of key nutrients is higher when they come from animal- vs plant-based foods. Meat reduction may also affect the balance between foods co-produced within the same animal production system. The objective was to assess the impact of introducing nutrient bioavailability and co-production links considerations on the dietary changes needed - especially regarding meat ‒ to improve diet sustainability. Diet optimization with linear and non-linear programming was used to design, for each gender, three modeled diets departing the least from the mean observed French diet (OBS) while reducing by at least 30% the diet-related environmental impacts (greenhouse gas emissions, eutrophication, acidification): i) in the nutrition-environment (NE) model, the fulfillment of recommended dietary allowances for all nutrients was imposed; ii) in the NE-bioavailability (NEB) model, nutritional adequacy was further ensured by accounting for iron, zinc, protein and provitamin A bioavailability; iii) in the NEB-co-production (NEB-CP) model, two links between co-produced animal foods (milk-beef and blood sausage-pork) were additionally included into the models by proportionally co-constraining their respective quantities. The price and environmental impacts of individual foods were assumed to be constant. 'Fruit and vegetables' and 'Starches' quantities increased in all modeled diets compared to OBS. In parallel, total meat and ruminant meat quantities decreased. Starting from 110g/d women's OBS diet (168g/d for men), total meat quantity decreased by 78%, 67% and 32% for women (68%, 66% and 62% for men) in NE, NEB and NEB-CP diets, respectively. Starting from 36g/d women's OBS diet (54g/d for men), ruminant meat quantity dropped severely by 84% and 87% in NE and NEB diets for women (80% and 78% for men), whereas it only decreased by 27% in NEB-CP diets (38% for men). The share of energy and proteins of animal origin was similar for the 3 modeled diets (approximately 1/5 of total energy, and 1/2 of protein) and lower than in OBS diet (approximately 1/3 of total energy, and 2/3 of protein). Decreasing meat content was strictly needed to achieve more sustainable diets for French adults, but the reduction was less severe when nutrient bioavailability and co-production links were taken into account.

  4. Digital encoding of cellular mRNAs enabling precise and absolute gene expression measurement by single-molecule counting.

    PubMed

    Fu, Glenn K; Wilhelmy, Julie; Stern, David; Fan, H Christina; Fodor, Stephen P A

    2014-03-18

    We present a new approach for the sensitive detection and accurate quantitation of messenger ribonucleic acid (mRNA) gene transcripts in single cells. First, the entire population of mRNAs is encoded with molecular barcodes during reverse transcription. After amplification of the gene targets of interest, molecular barcodes are counted by sequencing or scored on a simple hybridization detector to reveal the number of molecules in the starting sample. Since absolute quantities are measured, calibration to standards is unnecessary, and many of the relative quantitation challenges such as polymerase chain reaction (PCR) bias are avoided. We apply the method to gene expression analysis of minute sample quantities and demonstrate precise measurements with sensitivity down to sub single-cell levels. The method is an easy, single-tube, end point assay utilizing standard thermal cyclers and PCR reagents. Accurate and precise measurements are obtained without any need for cycle-to-cycle intensity-based real-time monitoring or physical partitioning into multiple reactions (e.g., digital PCR). Further, since all mRNA molecules are encoded with molecular barcodes, amplification can be used to generate more material for multiple measurements and technical replicates can be carried out on limited samples. The method is particularly useful for small sample quantities, such as single-cell experiments. Digital encoding of cellular content preserves true abundance levels and overcomes distortions introduced by amplification.

  5. Ethanol-induced impairment of polyamine homeostasis – A potential cause of neural tube defect and intrauterine growth restriction in fetal alcohol syndrome

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Haghighi Poodeh, Saeid, E-mail: saeid.haghighi@oulu.fi; Medical Research Center, Oulu University Hospital, Oulu; Alhonen, Leena

    Highlights: • Polyamine pools in embryonic and extraembryonic tissues are developmentally regulated. • Alcohol administration perturbs polyamine levels in the tissues with various patterns. • Total absence of polyamines in the embryo head at 9.5 dpc is critical for development. • The deficiency is associated with reduction in endothelial cell sprouting in the head. • Retarded migration of neural crest cells may cause development of neural tube defect. - Abstract: Introduction: Polyamines play a fundamental role during embryogenesis by regulating cell growth and proliferation and by interacting with RNA, DNA and protein. The polyamine pools are regulated by metabolism andmore » uptake from exogenous sources. The use of certain inhibitors of polyamine synthesis causes similar defects to those seen in alcohol exposure e.g. retarded embryo growth and endothelial cell sprouting. Methods: CD-1 mice received two intraperitoneal injections of 3 g/kg ethanol at 4 h intervals 8.75 days post coitum (dpc). The fetal head, trunk, yolk sac and placenta were collected at 9.5 and 12.5 dpc and polyamine concentrations were determined. Results: No measurable quantity of polyamines could be detected in the embryo head at 9.5 dpc, 12 h after ethanol exposure. Putrescine was not detectable in the trunk of the embryo at that time, whereas polyamines in yolk sac and placenta were at control level. Polyamine deficiency was associated with slow cell growth, reduction in endothelial cell sprouting, an altered pattern of blood vessel network formation and consequently retarded migration of neural crest cells and growth restriction. Discussion: Our results indicate that the polyamine pools in embryonic and extraembryonic tissues are developmentally regulated. Alcohol administration, at the critical stage, perturbs polyamine levels with various patterns, depending on the tissue and its developmental stage. The total absence of polyamines in the embryo head at 9.5 dpc may explain why this stage is so vulnerable to the development of neural tube defect, and growth restriction, the findings previously observed in fetal alcohol syndrome.« less

  6. Evaluation of electrical fields inside a biological structure.

    PubMed Central

    Drago, G. P.; Ridella, S.

    1982-01-01

    A digital computer simulation has been carried out of exposure of a cell, modelled as a multilayered spherical structure, to an alternating electrical field. Electrical quantities of possible biological interest can be evaluated everywhere inside the cell. A strong frequency selective behaviour in the range 0-10 MHz has been obtained. PMID:6279135

  7. Gene screening of Wharton's jelly derived stem cells.

    PubMed

    Mechiche Alami, S; Velard, F; Draux, F; Siu Paredes, F; Josse, J; Lemaire, F; Gangloff, S C; Graesslin, O; Laurent-Maquin, D; Kerdjoudj, H

    2014-01-01

    Stem cells are the most powerful candidate for the treatment of various diseases. Suitable stem cell source should be harvested with minimal invasive procedure, found in great quantity, and transplanted with no risk of immune response and tumor formation. Fetal derived stem cells have been introduced as an excellent alternative to adult and embryonic stem cells use, but unfortunately, their degree of "stemness" and molecular characterization is still unclear. Several studies have been performed deciphering whether fetal stem cells meet the needs of regenerative medicine. We believe that a transcriptomic screening of Wharton's jelly stem cells will bring insights on cell population features.

  8. Production of minimally disturbed synchronous cultures of hematopoietic cells

    NASA Technical Reports Server (NTRS)

    Thornton, Maureen; Eward, Kathryn Leigh; Helmstetter, Charles E.; Edward, K. L. (Principal Investigator)

    2002-01-01

    A method is describedforproducing sizable quantities of synchronously dividing, minimally disturbed mammalian cells. Cultures were grown immobilized on surfaces such that cell division within the population resulted in the continuous release of synchronous newborn cells. As judged by the quality and duration of synchronous growth, cell size distributions, and DNA compositions, newborn mouse L1210 cells grew with a very high level of synchrony without overt evidence of growth disturbances. The technology should be applicable to a variety of hematopoietic cells, as evidenced by similar results with human MOLT-4 and U937 cell lines.

  9. System and method for conditioning a hardwood pulp liquid hydrolysate

    DOEpatents

    Waite, Darrell M; Arnold, Richard; St. Pierre, James; Pendse, Hemant P; Ceckler, William H

    2013-12-17

    A system and method for hardwood pulp liquid hydrolysate conditioning includes a first evaporator receives a hardwood mix extract and outputting a quantity of vapor and extract. A hydrolysis unit receives the extract, hyrolyzes and outputs to a lignin separation device, which separates and recovers a quantity of lignin. A neutralization device receives extract from the lignin separation device and a neutralizing agent, producing a mixture of solid precipitate and a fifth extract. The solid precipitate is removed from the fifth extract. A second evaporator removes a quantity of acid from the fifth extract in a vapor form. This vapor may be recycled to improve total acid recovery or discarded. A desalination device receives the diluted extract, separates out some of the acid and salt and outputs a desalinated solution.

  10. The role of astrocytes in amyloid production and Alzheimer's disease

    PubMed Central

    Frost, Georgia R.

    2017-01-01

    Alzheimer's disease (AD) is marked by the presence of extracellular amyloid beta (Aβ) plaques, intracellular neurofibrillary tangles (NFTs) and gliosis, activated glial cells, in the brain. It is thought that Aβ plaques trigger NFT formation, neuronal cell death, neuroinflammation and gliosis and, ultimately, cognitive impairment. There are increased numbers of reactive astrocytes in AD, which surround amyloid plaques and secrete proinflammatory factors and can phagocytize and break down Aβ. It was thought that neuronal cells were the major source of Aβ. However, mounting evidence suggests that astrocytes may play an additional role in AD by secreting significant quantities of Aβ and contributing to overall amyloid burden in the brain. Astrocytes are the most numerous cell type in the brain, and therefore even minor quantities of amyloid secretion from individual astrocytes could prove to be substantial when taken across the whole brain. Reactive astrocytes have increased levels of the three necessary components for Aβ production: amyloid precursor protein, β-secretase (BACE1) and γ-secretase. The identification of environmental factors, such as neuroinflammation, that promote astrocytic Aβ production, could redefine how we think about developing therapeutics for AD. PMID:29237809

  11. Silicon Dioxide Thin Film Mediated Single Cell Nucleic Acid Isolation

    PubMed Central

    Bogdanov, Evgeny; Dominova, Irina; Shusharina, Natalia; Botman, Stepan; Kasymov, Vitaliy; Patrushev, Maksim

    2013-01-01

    A limited amount of DNA extracted from single cells, and the development of single cell diagnostics make it necessary to create a new highly effective method for the single cells nucleic acids isolation. In this paper, we propose the DNA isolation method from biomaterials with limited DNA quantity in sample, and from samples with degradable DNA based on the use of solid-phase adsorbent silicon dioxide nanofilm deposited on the inner surface of PCR tube. PMID:23874571

  12. Development of an Ammonium Sulfate DNA Extraction Method for Obtaining Amplifiable DNA in a Small Number of Cells and Its Application to Clinical Specimens

    PubMed Central

    Oh, Seo Young; Kim, Wook Youn; Hwang, Tae Sook; Han, Hye Seung; Lim, So Dug; Kim, Wan Seop

    2013-01-01

    DNA extraction from microdissected cells has become essential for handling clinical specimens with advances in molecular pathology. Conventional methods have limitations for extracting amplifiable DNA from specimens containing a small number of cells. We developed an ammonium sulfate DNA extraction method (A) and compared it with two other methods (B and C). DNA quality and quantity, β-globin amplification, and detectability of two cancer associated gene mutations were evaluated. Method A showed the best DNA yield, particularly when the cell number was very low. Amplification of the β-globin gene using DNA from the SNU 790 cell line and papillary thyroid carcinoma (PTC) cells extracted with Method A demonstrated the strongest band. BRAF V600E mutation analysis using ethanol-fixed PTC cells from a patient demonstrated both a “T” peak increase and an adjacent “A” peak decrease when 25 and 50 cells were extracted, whereas mutant peaks were too low to be analyzed using the other two methods. EGFR mutation analysis using formalin-fixed paraffin-embedded lung cancer tissues demonstrated a mutant peak with Method A, whereas the mutant peak was undetectable with Methods B or C. Method A yielded the best DNA quantity and quality with outstanding efficiency, particularly when paucicellular specimens were used. PMID:23691506

  13. Limited elimination of two viruses by cryotherapy of pelargonium apices related to virus distribution.

    PubMed

    Gallard, A; Mallet, R; Chevalier, M; Grapin, A

    2011-01-01

    The possibility of eradicating the pelargonium flower break virus (PFBV) and pelargonium line pattern virus (PLPV) by cryotherapy of axillary shoot apices was investigated using five Pelargonium cultivars. Viruses were detected by DAS-ELISA and their location was determined by immunolocalization. Apex culture did not permit elimination of PFBV and only 15 percent regenerated plants of 'Stellar Artic' cultivar were ELISA PLPV-negative. Plants regenerated from cryotherapy-treated apices were tested by DAS-ELISA after a 3-month in vitro culture period. Viruses were not detected in 25 percent and 50 percent of the plants tested for PFBV and PLPV, respectively. However, immunolocalization carried out on apices originating from cryopreserved shoot tips sampled from DAS-ELISA negative plants showed that they were still virus-infected. Using immunolocalization, PFBV and PLPV could be detected in Pelargonium apices, even in the meristematic dome. However, viral particles were more numerous in basal zone cells than in meristematic cells. Our results demonstrate that PFBV and PLPV are present within meristematic cells and that cryopreservation can partly reduce the quantity of these viruses in Pelargonium plants but not eliminate them totally. Additional knowledge on localization and behaviour of viruses during cryopreservation is essential to optimize cryotherapy and plant genetic resource management.

  14. Biosorption of Strontium from Simulated Nuclear Wastewater by Scenedesmus spinosus under Culture Conditions: Adsorption and Bioaccumulation Processes and Models

    PubMed Central

    Liu, Mingxue; Dong, Faqin; Kang, Wu; Sun, Shiyong; Wei, Hongfu; Zhang, Wei; Nie, Xiaoqin; Guo, Yuting; Huang, Ting; Liu, Yuanyuan

    2014-01-01

    Algae biosorption is an ideal wastewater treatment method when coupled with algae growth and biosorption. The adsorption and bioaccumulation of strontium from simulated nuclear wastewater by Scenedesmus spinosus were investigated in this research. One hundred mL of cultured S. spinosus cells with a dry weight of 1.0 mg in simulated nuclear wastewater were used to analyze the effects on S. spinosus cell growth as well as the adsorption and bioaccumulation characters under conditions of 25 ± 1 °C with approximately 3,000 lux illumination. The results showed that S. spinosus had a highly selective biosorption capacity for strontium, with a maximum bioremoval ratio of 76%. The adsorbed strontium ion on cell walls was approximately 90% of the total adsorbed amount; the bioaccumulation in the cytoplasm varied by approximately10%. The adsorption quantity could be described with an equilibrium isotherm. The pseudo-second-order kinetic model suggested that adsorption was the rate-limiting step of the biosorption process. A new bioaccumulation model with three parameters was proposed and could give a good fit with the experiment data. The results suggested that S. spinosus may be a potential biosorbent for the treatment of nuclear wastewater in culture conditions. PMID:24919131

  15. 9 CFR 381.121 - Quantity of contents.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ..., in distinct contrast to other matter on the container, and shall be declared in accordance with the... totally with impervious packaging material and is packed with a usable medium. (2) The statement shall be...

  16. 9 CFR 381.121 - Quantity of contents.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ..., in distinct contrast to other matter on the container, and shall be declared in accordance with the... totally with impervious packaging material and is packed with a usable medium. (2) The statement shall be...

  17. 14 CFR 125.9 - Definitions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... OR MORE; AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT General § 125.9 Definitions. (a) For the... total quantity of hydraulic fluid. (c) For the purposes of this part, maximum zero fuel weight means the...

  18. 14 CFR 125.9 - Definitions.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... OR MORE; AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT General § 125.9 Definitions. (a) For the... total quantity of hydraulic fluid. (c) For the purposes of this part, maximum zero fuel weight means the...

  19. 14 CFR 125.9 - Definitions.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... OR MORE; AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT General § 125.9 Definitions. (a) For the... total quantity of hydraulic fluid. (c) For the purposes of this part, maximum zero fuel weight means the...

  20. 14 CFR 125.9 - Definitions.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... OR MORE; AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT General § 125.9 Definitions. (a) For the... total quantity of hydraulic fluid. (c) For the purposes of this part, maximum zero fuel weight means the...

  1. 14 CFR 125.9 - Definitions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... OR MORE; AND RULES GOVERNING PERSONS ON BOARD SUCH AIRCRAFT General § 125.9 Definitions. (a) For the... total quantity of hydraulic fluid. (c) For the purposes of this part, maximum zero fuel weight means the...

  2. Finite horizon EOQ model for non-instantaneous deteriorating items with price and advertisement dependent demand and partial backlogging under inflation

    NASA Astrophysics Data System (ADS)

    Palanivel, M.; Uthayakumar, R.

    2015-07-01

    This paper deals with an economic order quantity (EOQ) model for non-instantaneous deteriorating items with price and advertisement dependent demand pattern under the effect of inflation and time value of money over a finite planning horizon. In this model, shortages are allowed and partially backlogged. The backlogging rate is dependent on the waiting time for the next replenishment. This paper aids the retailer in minimising the total inventory cost by finding the optimal interval and the optimal order quantity. An algorithm is designed to find the optimum solution of the proposed model. Numerical examples are given to demonstrate the results. Also, the effect of changes in the different parameters on the optimal total cost is graphically presented and the implications are discussed in detail.

  3. Electric Power Quarterly, July-September 1984

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Not Available

    1985-01-01

    The Electric Power Quarterly (EPQ) provides electric utilities' plant-level information about the cost, quantity, and quality of fossil fuel receipts, net generation, fuel consumption, and fuel stocks. The EPQ contains monthly data and quarterly totals for the reporting quarter. In this report, data collected on Form EIA-759 regarding electric utilities' net generation, fuel consumption, and fuel stocks are presented on a plant-by-plant basis. In addition, quantity, cost, and quality of fossil fuel receipts collected on the Federal Energy Regulatory Commission (FERC) Form 423 are presented on a plant-by-plant basis.

  4. Electric Power Quarterly, October-December 1984

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Not Available

    1985-04-01

    The Electric Power Quarterly (EPQ) provides electric utilities' plant-level information about the cost, quantity, and quality of fossil fuel receipts, net generation, fuel consumption, and fuel stocks. The EPQ contains monthly data and quarterly totals for the reporting quarter. In this report, data collected on Form EIA-759 regarding electric utilities' net generation, fuel consumption, and fuel stocks are presented on a plant-by-plant basis. In addition, quantity, cost, and quality of fossil fuel receipts collected on the Federal Energy Regulatory Commission (FERC) Form 423 are presented on a plant-by-plant basis.

  5. An EOQ model for weibull distribution deterioration with time-dependent cubic demand and backlogging

    NASA Astrophysics Data System (ADS)

    Santhi, G.; Karthikeyan, K.

    2017-11-01

    In this article we introduce an economic order quantity model with weibull deterioration and time dependent cubic demand rate where holding costs as a linear function of time. Shortages are allowed in the inventory system are partially and fully backlogging. The objective of this model is to minimize the total inventory cost by using the optimal order quantity and the cycle length. The proposed model is illustrated by numerical examples and the sensitivity analysis is performed to study the effect of changes in parameters on the optimum solutions.

  6. Construction Productivity Advancement Research (CPAR) Program: Improved Materials and Processes for Sealing and Resealing Joints in Portland Cement Concrete Pavements - Field Evaluation

    DTIC Science & Technology

    1993-10-01

    sealant was determined by noting the type and number of defects each sealant incurred. Figure 4 provides a sample evaluation sheet used dur- ing the field...was conducted by visually inspect- ing the mater~al for defects . If any defects were noted, the type of defect was described and the quant~ty of that... defect was measured. The quantity of the defect was dividted by the total quantity of sealant and the result reported as percent defect . Adhesion and

  7. Analysis and metabolic engineering of lipid-linked oligosaccharides in glycosylation-deficient CHO cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Jones, Meredith B., E-mail: mbauman7@jhu.edu; Tomiya, Noboru, E-mail: ntomiya1@jhu.edu; Betenbaugh, Michael J., E-mail: beten@jhu.edu

    2010-04-23

    Glycosylation-deficient Chinese Hamster Ovary (CHO) cell lines can be used to expand our understanding of N-glycosylation pathways and to study Congenital Disorders of Glycosylation, diseases caused by defects in the synthesis of N-glycans. The mammalian N-glycosylation pathway involves the step-wise assembly of sugars onto a dolichol phosphate (P-Dol) carrier, forming a lipid-linked oligosaccharide (LLO), followed by the transfer of the completed oligosaccharide onto the protein of interest. In order to better understand how deficiencies in this pathway affect the availability of the completed LLO donor for use in N-glycosylation, we used a non-radioactive, HPLC-based assay to examine the intermediates inmore » the LLO synthesis pathway for CHO-K1 cells and for three different glycosylation-deficient CHO cell lines. B4-2-1 cells, which have a mutation in the dolichol phosphate-mannose synthase (DPM2) gene, accumulated LLO with the structure Man{sub 5}GlcNAc{sub 2}-P-P-Dol, while MI8-5 cells, which lack glucosyltransferase I (ALG6) activity, accumulated Man{sub 9}GlcNAc{sub 2}-P-P-Dol. CHO-K1 and MI5-4 cells both produced primarily the complete LLO, Glc{sub 3}Man{sub 9}GlcNAc{sub 2}-P-P-Dol, though the relative quantity was lower in MI5-4. MI5-4 cells have reduced hexokinase activity which could affect the availability of many of the substrates required for LLO synthesis and, consequently, impair production of the final LLO donor. Increasing hexokinase activity by overexpressing hexokinase II in MI5-4 caused a decrease in the relative quantities of the incomplete LLO intermediates from Man{sub 5}GlcNAc{sub 2}-PP-Dol through Glc{sub 1}Man{sub 9}GlcNAc{sub 2}-PP-Dol, and an increase in the relative quantity of the final LLO donor, Glc{sub 3}Man{sub 9}GlcNAc{sub 2}-P-P-Dol. This study suggests that metabolic engineering may be a useful strategy for improving LLO availability for use in N-glycosylation.« less

  8. Use of and Self-Perceived Need for Assistive Devices in Individuals with Disabilities in Taiwan

    PubMed Central

    Yeung, Kwok-Tak; Lin, Chung-Hui; Teng, Ya-Ling; Chen, Fen-Fen; Lou, Shu-Zon; Chen, Chiung-Ling

    2016-01-01

    Assistive devices (ADs) can help individuals with disabilities achieve greater independence, and it can enhance the quality of their lives. This study investigated the use of and self-perceived need for ADs in individuals with disabilities, and determined the influence of gender, age as well as type and degree of disability on the use of and self-perceived need for ADs. This descriptive study utilized a cross-sectional survey design with a convenience sample of participants. A total of 1018 subjects with disabilities who visited an exhibition of assistive technology and two ADs research and development centers completed a questionnaires either by themselves or via a caregiver who completed the questionnaire on behalf of the subject or via interviewers trained specifically for this study. The Mann-Whitney U test and Kruskal-Wallis test were used to determine the influence of participant characteristics on the use of ADs. The results showed that 77.2% and 83.3% of the participants reported that they used and needed AD(s) to engage in activities of daily living. The mean quantity of the use of and self-perceived need for total types of ADs were 3.0 and 5.3, respectively. Participants with different disabilities reported different percentages of the use of various types of ADs. No difference was found between genders and among the age groups in the use of quantity of ADs. Individuals with different types and degrees of disability used different quantities of ADs. Participants with physical, visual and multiple disabilities used significantly more ADs compared to participants with intellectual disability. The total quantity of ADs used increased significantly with increased severity of disability. The mean use of assistive devices was lower compared to the mean need of individuals with disabilities. Further study is required to determine why patients feel the need for but not currently use a specific assistive device. PMID:27023276

  9. Gas chromatograph sample-transfer valve

    NASA Technical Reports Server (NTRS)

    Wang, W. S.; Wright, H. W., Jr.

    1971-01-01

    Slide-type gate valve incorporates sampling volume and transfer passageway for guiding a metered quantity of gas from pressurized test cell to gas chromatograph. Gate is moved by pneumatic bellows-type actuator.

  10. Multi-scale modeling of APC and [Formula: see text]-catenin regulation in the human colonic crypt.

    PubMed

    Emerick, Brooks; Schleiniger, Gilberto; Boman, Bruce M

    2018-06-01

    Stem cell renewal and differentiation in the human colonic crypt are linked to the [Formula: see text]-catenin pathway. The spatial balance of Wnt factors in proliferative cells within the crypt maintain an appropriate level of cellular reproduction needed for normal crypt homeostasis. Mutational events at the gene level are responsible for deregulating the balance of Wnt factors along the crypt, causing an overpopulation of proliferative cells, a loss of structure of the crypt domain, and the initiation of colorectal carcinomas. We formulate a PDE model describing cell movement and reproduction in a static crypt domain. We consider a single cell population whose proliferative capabilities are determined by stemness, a quantity defined by intracellular levels of adenomatous polyposis coli (APC) scaffold protein and [Formula: see text]-catenin. We fit APC regulation parameters to biological data that describe normal protein gradients in the crypt. We also fit cell movement and protein flux parameters to normal crypt characteristics such as renewal time, total cell count, and proportion of proliferating cells. The model is used to investigate abnormal crypt dynamics when subjected to a diminished APC gradient, a scenario synonymous to mutations in the APC gene. We find that a 25% decrease in APC synthesis leads to a fraction of 0.88 proliferative, which is reflective of normal-appearing FAP crypts. A 50% drop in APC activity yields a fully proliferative crypt showing a doubling of the level of stemness, which characterizes the initial stages of colorectal cancer development. A sensitivity analysis of APC regulation parameters shows the perturbation of factors that is required to restore crypt dynamics to normal in the case of APC mutations.

  11. Synergistic cytotoxicity and mechanism of caffeine and lysozyme on hepatoma cell line HepG2

    NASA Astrophysics Data System (ADS)

    Yang, Hongchao; Li, Jingjuan; Cui, Lin; Ren, Yanqing; Niu, Liying; Wang, Xinguo; Huang, Yun; Cui, Lijian

    2018-03-01

    The influences of caffeine, lysozyme and the joint application of them on the hepatoma cell line HepG2 proliferation inhibition and cell apoptosis were observed by 3-(4, 5-dimethyl-2-thiazyl)-2, 5-diphenyl-2H-tetrazolium bromide assay and Hoechst 33342, which showed the proliferation inhibition rate of the joint application on HepG2 cells was 47.21%, significantly higher than caffeine or lysozyme, and the joint application promoted the apoptosis of HepG2 cells obviously. Van't Hoff classical thermodynamics formula, the Föster theory of non-radiation energy transfer and fluorescence phase diagram were used to manifest that the process of lysozyme binding to caffeine followed a two-state model, which was spontaneous at low temperature driven by enthalpy change, and the predominant intermolecular force was hydrogen bonding or Van der Waals force to stabilize caffeine-lysozyme complex with the distance 5.86 nm. The attenuated total reflection-Fourier transform infrared spectra indicated that caffeine decreased the relative contents of α-helix and β-turn, which inferred the structure of lysozyme tended to be "loose". Synchronous fluorescence spectra and ultraviolet spectra supported the above conclusion. The amino acid residues in the cleft of lysozyme were exposed and electropositivity was increased attributing to the loose structure, which were conducive to increasing caffeine concentration on the HepG2 cell surface by electrostatic interaction to show synergistic effect. The great quantities of microvilli on the liver cancer cell membrane surface, is beneficial for the lysozyme-caffeine compound to aggregate on cell surface to increase the concentration of caffeine to play stronger physiological role by electrostatic effect.

  12. Pretreatment with IL-1β enhances proliferation and chondrogenic potential of synovium-derived mesenchymal stem cells.

    PubMed

    Matsumura, Etsuko; Tsuji, Kunikazu; Komori, Keiichiro; Koga, Hideyuki; Sekiya, Ichiro; Muneta, Takeshi

    2017-02-01

    Synovial mesenchymal stem cells (MSCs) are an attractive cell source for cartilage regeneration because of their high proliferative ability and chondrogenic potential. We have performed clinical trials using synovial MSCs to regenerate articular cartilage. To achieve good clinical outcomes for cell transplantation therapy, it is important to control both quantity (cell number) and quality (pluripotency or chondrogenic potential) of the cells for transplantation. Interleukin (IL)-1β is a pro-inflammatory cytokine with significant pro-proliferative potential for mesenchymal cells. However, the effects of IL-1β on synovial MSCs remain unknown. We investigated the effects of pretreatment with IL-1β on synovial MSCs. Human synovial tissue was harvested during total knee arthroplasty. Nucleated cells were plated and cultured in the absence or presence of IL-1β at 10 -13 , 10 -12 , 10 -11 , 10 -10 , 10 -9 or 10 -8  g/mL for 14 days. The number of synovial MSCs increased in a concentration-dependent manner. When cultured for 21 days in chondrogenic medium after pretreatment with 10 -8   g/mL IL-1β, pellet aggregation was observed, whereas pretreatment with 10 -12 , 10 -11 or 10 -10  g/mL IL-1β significantly increased the weight of cartilage pellets (P <0.01). Surface markers for adhesion ability and pluripotency were reduced with high concentrations of IL-1β. IL-6 and IL-8 expression increased, but no changes in the expression level of growth factors were indicated by cytokine array. We have demonstrated that pretreatment of IL-1β increased the proliferation and chondrogenic potential of synovial MSCs, which may promote the regenerative potential of synovial MSCs. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  13. Differential nuclear staining assay for high-throughput screening to identify cytotoxic compounds.

    PubMed

    Lema, Carolina; Varela-Ramirez, Armando; Aguilera, Renato J

    As large quantities of novel synthetic molecules continue to be generated there is a challenge to identify therapeutic agents with cytotoxic activity. Here we introduce a Differential Nuclear Staining (DNS) assay adapted to live-cell imaging for high throughput screening (HTS) that utilizes two fluorescent DNA intercalators, Hoechst 33342 and Propidium iodide (PI). Since Hoechst can readily cross cell membranes to stain DNA of living and dead cells, it was used to label the total number of cells. In contrast, PI only enters cells with compromised plasma membranes, thus selectively labeling dead cells. The DNS assay was successfully validated by utilizing well known cytotoxic agents with fast or slow cytotoxic activities. The assay was found to be suitable for HTS with Z' factors ranging from 0.86 to 0.60 for 96 and 384-well formats, respectively. Furthermore, besides plate-to-plate reproducibility, assay quality performance was evaluated by determining ratios of signal-to-noise and signal-to-background, as well as coefficient of variation, which resulted in adequate values and validated the assay for HTS initiatives. As proof of concept, eighty structurally diverse compounds from a small molecule library were screened in a 96-well plate format using the DNS assay. Using this DNS assay, six hits with cytotoxic properties were identified and all of them were also successfully identified by using the commercially available MTS assay (CellTiter 96® Cell Proliferation Assay). In addition, the DNS and a flow cytometry assay were used to validate the activity of the cytotoxic compounds. The DNS assay was also used to generate dose-response curves and to obtain CC 50 values. The results indicate that the DNS assay is reliable and robust and suitable for primary and secondary screens of compounds with potential cytotoxic activity.

  14. Differential nuclear staining assay for high-throughput screening to identify cytotoxic compounds

    PubMed Central

    LEMA, Carolina; VARELA-RAMIREZ, Armando; AGUILERA, Renato J.

    2016-01-01

    As large quantities of novel synthetic molecules continue to be generated there is a challenge to identify therapeutic agents with cytotoxic activity. Here we introduce a Differential Nuclear Staining (DNS) assay adapted to live-cell imaging for high throughput screening (HTS) that utilizes two fluorescent DNA intercalators, Hoechst 33342 and Propidium iodide (PI). Since Hoechst can readily cross cell membranes to stain DNA of living and dead cells, it was used to label the total number of cells. In contrast, PI only enters cells with compromised plasma membranes, thus selectively labeling dead cells. The DNS assay was successfully validated by utilizing well known cytotoxic agents with fast or slow cytotoxic activities. The assay was found to be suitable for HTS with Z′ factors ranging from 0.86 to 0.60 for 96 and 384-well formats, respectively. Furthermore, besides plate-to-plate reproducibility, assay quality performance was evaluated by determining ratios of signal-to-noise and signal-to-background, as well as coefficient of variation, which resulted in adequate values and validated the assay for HTS initiatives. As proof of concept, eighty structurally diverse compounds from a small molecule library were screened in a 96-well plate format using the DNS assay. Using this DNS assay, six hits with cytotoxic properties were identified and all of them were also successfully identified by using the commercially available MTS assay (CellTiter 96® Cell Proliferation Assay). In addition, the DNS and a flow cytometry assay were used to validate the activity of the cytotoxic compounds. The DNS assay was also used to generate dose-response curves and to obtain CC50 values. The results indicate that the DNS assay is reliable and robust and suitable for primary and secondary screens of compounds with potential cytotoxic activity. PMID:27042697

  15. Pigments, size and distribution of Synechococcus spp. in the Black Sea

    NASA Astrophysics Data System (ADS)

    Uysal, Zahit

    2000-03-01

    Pigments, size and distribution of Phycoerythrin-containing unicellular cyanobacteria Synechococcus spp. within the euphotic zone were studied for the first time in April-May 1994 in the western and southwestern Black Sea by epifluorescence microscopy and flow-cytometry. Synechococcus was present in varying quantities at every station and depth studied. Surface spatial distribution of Synechococcus revealed that cells were much more abundant in offshore waters than near coastal regions under the direct influence of the Danube river. Minimum and maximum cell concentrations ranged between 9×10 2 and 1.45×10 5 cells/ml at the surface, between 2×10 3 and 1.23×10 5 cells/ml at the chlorophyll sub-maximum layer, and between 1.3×10 2 and 3.5×10 2 at the nitrite maximum layer. Cells at the chlorophyll sub-maximum layer (based on in-situ fluorometer readings) fluoresce brighter and longer than the ones at the surface and lower depths. Spectral properties of chromophore pigment types of total 64 clonal isolates from different depths down to the lower layer of the euphotic zone (˜60 m) in the southern Black Sea coast revealed that all have type 2 phycoerythrobilin in common, lacking in phycourobilin. In vivo fluorescence emission maxima for the phycoerythrobilin were about the same (˜578 nm) for all isolates. All isolates examined showed in vivo absorption maxima at between 435 and 442 nm and at about 681 nm due to chlorophyll- a. Based on the flow cytometer mean forward light scatter data for size distribution, it could be concluded that cells at the surface mixed layer (0-10 m) were larger in cell size than the cells at lower depths (20-60 m).

  16. Sleep Quality in Medical Students; the Impact of Over-Use of Mobile Cell-Phone and Social Networks.

    PubMed

    Mohammadbeigi, Abolfazl; Absari, Rozita; Valizadeh, Farzaneh; Saadati, Mohammadreza; Sharifimoghadam, Soroush; Ahmadi, Ali; Mokhtari, Mohsen; Ansari, Hossein

    2016-01-01

    Poor sleep quality is closely associated with lifestyle habits including use of mobile cell-phones.This study aimed to identify the relationship between sleep quality due to abuse in mobile cell-phones and engagement in social networks. This cross-sectional study was conducted on 380 undergraduate students selected by proportional stratified sampling in Qom, Iran in 2015. Data were collected by two standard questionnaire including Cell-Phone Over-Use Scale (COS) and Pittsburgh sleep quality questionnaire beside the status of usage in cell-phone social networks. T-test, chi-square, Pearson correlation coefficient and multivariate logistic regression were used in data analysis. The mean age of participants was 21.8 ±3.2 yr, 69.1% were female, and 11.7% were married. The mean of COS and sleep quality scores were 48.18 ±17.5 and 5.38 ±2.31, respectively. The prevalence of over-use of cell phone was 10.7% (CI 0.95; 8.8%, 12.6%) and the prevalence of poor sleep quality was 61.7% (CI 0.95; 57.1%, 66.3%). The mean of all aspects and total score of sleep quality showed a direct significant association by cell-phone addiction score except sleep duration score that was inversely. Based on multivariate analysis affected to cell-phone addiction, being male gender and studying in general physician level are the most important predictors of poor sleep quality. Over use of internet and social networks via smart phones is related to poor sleep quality and quantity. Predefined sport programs, educational, cultural, and interesting entertainment are the essential needs for all medical students. These interventions are more important especially for male students who have longer educational.

  17. 49 CFR 172.202 - Description of hazardous material on shipping papers.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...) Bulk packages, provided some indication of the total quantity is shown, for example, “1 cargo tank” or... cylinders.” (C) Packages containing only residue. (6) For transportation by aircraft, the total net mass per package, must be shown unless a gross mass is indicated in Columns (9A) or (9B) of the § 172.101 table in...

  18. 49 CFR 172.202 - Description of hazardous material on shipping papers.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...) Bulk packages, provided some indication of the total quantity is shown, for example, “1 cargo tank” or... cylinders.” (C) Packages containing only residue. (6) For transportation by aircraft, the total net mass per package, must be shown unless a gross mass is indicated in Columns (9A) or (9B) of the § 172.101 table in...

  19. 49 CFR 172.202 - Description of hazardous material on shipping papers.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...) Bulk packages, provided some indication of the total quantity is shown, for example, “1 cargo tank” or... cylinders.” (C) Packages containing only residue. (6) For transportation by aircraft, the total net mass per package, must be shown unless a gross mass is indicated in Columns (9A) or (9B) of the § 172.101 table in...

  20. Microbial Contents of Vacuum Cleaner Bag Dust and Emitted Bioaerosols and Their Implications for Human Exposure Indoors

    PubMed Central

    Veillette, Marc; Knibbs, Luke D.; Pelletier, Ariane; Charlebois, Remi; Blais Lecours, Pascale; He, Congrong; Morawska, Lidia

    2013-01-01

    Vacuum cleaners can release large concentrations of particles, both in their exhaust air and from resuspension of settled dust. However, the size, variability, and microbial diversity of these emissions are unknown, despite evidence to suggest they may contribute to allergic responses and infection transmission indoors. This study aimed to evaluate bioaerosol emission from various vacuum cleaners. We sampled the air in an experimental flow tunnel where vacuum cleaners were run, and their airborne emissions were sampled with closed-face cassettes. Dust samples were also collected from the dust bag. Total bacteria, total archaea, Penicillium/Aspergillus, and total Clostridium cluster 1 were quantified with specific quantitative PCR protocols, and emission rates were calculated. Clostridium botulinum and antibiotic resistance genes were detected in each sample using endpoint PCR. Bacterial diversity was also analyzed using denaturing gradient gel electrophoresis (DGGE), image analysis, and band sequencing. We demonstrated that emission of bacteria and molds (Penicillium/Aspergillus) can reach values as high as 1E5 cell equivalents/min and that those emissions are not related to each other. The bag dust bacterial and mold content was also consistent across the vacuums we assessed, reaching up to 1E7 bacterial or mold cell equivalents/g. Antibiotic resistance genes were detected in several samples. No archaea or C. botulinum was detected in any air samples. Diversity analyses showed that most bacteria are from human sources, in keeping with other recent results. These results highlight the potential capability of vacuum cleaners to disseminate appreciable quantities of molds and human-associated bacteria indoors and their role as a source of exposure to bioaerosols. PMID:23934489

  1. Current status of treating neurodegenerative disease with induced pluripotent stem cells.

    PubMed

    Pen, A E; Jensen, U B

    2017-01-01

    Degenerative diseases of the brain have proven challenging to treat, let alone cure. One of the treatment options is the use of stem cell therapy, which has been under investigation for several years. However, treatment with stem cells comes with a number of drawbacks, for instance the source of these cells. Currently, a number of options are tested to produce stem cells, although the main issues of quantity and ethics remain for most of them. Over recent years, the potential of induced pluripotent stem cells (iPSCs) has been widely investigated and these cells seem promising for production of numerous different tissues both in vitro and in vivo. One of the major advantages of iPSCs is that they can be made autologous and can provide a sufficient quantity of cells by culturing, making the use of other stem cell sources unnecessary. As the first descriptions of iPSC production with the transcription factors Sox2, Klf4, Oct4 and C-Myc, called the Yamanaka factors, a variety of methods has been developed to convert somatic cells from all germ layers to pluripotent stem cells. Improvement of these methods is necessary to increase the efficiency of reprogramming, the quality of pluripotency and the safety of these cells before use in human trials. This review focusses on the current accomplishments and remaining challenges in the production and use of iPSCs for treatment of neurodegenerative diseases of the brain such as Alzheimer's disease and Parkinson's disease. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  2. Bone Marrow Aspirate Concentrate for Cartilage Defects of the Knee: From Bench to Bedside Evidence.

    PubMed

    Cotter, Eric J; Wang, Kevin C; Yanke, Adam B; Chubinskaya, Susan

    2018-04-01

    Objective To critically evaluate the current basic science, translational, and clinical data regarding bone marrow aspirate concentrate (BMAC) in the setting of focal cartilage defects of the knee and describe clinical indications and future research questions surrounding the clinical utility of BMAC for treatment of these lesions. Design A literature search was performed using the PubMed and Ovid MEDLINE databases for studies in English (1980-2017) using keywords, including ["bone marrow aspirate" and "cartilage"], ["mesenchymal stem cells" and "cartilage"], and ["bone marrow aspirate" and "mesenchymal stem cells" and "orthopedics"]. A total of 1832 articles were reviewed by 2 independent authors and additional literature found through scanning references of cited articles. Results BMAC has demonstrated promising results in the clinical application for repair of chondral defects as an adjuvant procedure or as an independent management technique. A subcomponent of BMAC, bone marrow derived-mesenchymal stem cells (MSCs) possess the ability to differentiate into cells important for osteogenesis and chondrogenesis. Modulation of paracrine signaling is perhaps the most important function of BM-MSCs in this setting. In an effort to increase the cellular yield, authors have shown the ability to expand BM-MSCs in culture while maintaining phenotype. Conclusions Translational studies have demonstrated good clinical efficacy of BMAC both concomitant with cartilage restoration procedures, at defined time points after surgery, and as isolated injections. Early clinical data suggests BMAC may help stimulate a more robust hyaline cartilage repair tissue response. Numerous questions remain regarding BMAC usage, including cell source, cell expansion, optimal pathology, and injection timing and quantity.

  3. Use of focused acoustics for cell disruption to provide ultra scale-down insights of microbial homogenization and its bioprocess impact--recovery of antibody fragments from rec E. coli.

    PubMed

    Li, Qiang; Aucamp, Jean P; Tang, Alison; Chatel, Alex; Hoare, Mike

    2012-08-01

    An ultra scale-down (USD) device that provides insight of how industrial homogenization impacts bioprocess performance is desirable in the biopharmaceutical industry, especially at the early stage of process development where only a small quantity of material is available. In this work, we assess the effectiveness of focused acoustics as the basis of an USD cell disruption method to mimic and study high-pressure, step-wise homogenization of rec Escherichia coli cells for the recovery of an intracellular protein, antibody fragment (Fab'). The release of both Fab' and of overall protein follows first-order reaction kinetics with respect to time of exposure to focused acoustics. The rate constant is directly proportional to applied electrical power input per unit volume. For nearly total protein or Fab' release (>99%), the key physical properties of the disruptate produced by focused acoustics, such as cell debris particle size distribution and apparent viscosity show good agreement with those for homogenates produced by high-pressure homogenization operated to give the same fractional release. The only key difference is observed for partial disruption of cells where focused acoustics yields a disruptate of lower viscosity than homogenization, evidently due to a greater extent of polynucleic acids degradation. Verification of this USD approach to cell disruption by high-pressure homogenization is achieved using USD centrifugation to demonstrate the same sedimentation characteristics of disruptates prepared using both the scaled-down focused acoustic and the pilot-scale homogenization methods for the same fraction of protein release. Copyright © 2012 Wiley Periodicals, Inc.

  4. Isolation of high-quality total RNA from leaves of Myrciaria dubia "CAMU CAMU".

    PubMed

    Gómez, Juan Carlos Castro; Reátegui, Alina Del Carmen Egoavil; Flores, Julián Torres; Saavedra, Roberson Ramírez; Ruiz, Marianela Cobos; Correa, Sixto Alfredo Imán

    2013-01-01

    Myrciaria dubia is a main source of vitamin C for people in the Amazon region. Molecular studies of M. dubia require high-quality total RNA from different tissues. So far, no protocols have been reported for total RNA isolation from leaves of this species. The objective of this research was to develop protocols for extracting high-quality total RNA from leaves of M. dubia. Total RNA was purified following two modified protocols developed for leaves of other species (by Zeng and Yang, and by Reid et al.) and one modified protocol developed for fruits of the studied species (by Silva). Quantity and quality of purified total RNA were assessed by spectrophotometric and electrophoretic analysis. Additionally, quality of total RNA was evaluated with reverse-transcription polymerase chain reaction (RT-PCR). With these three modified protocols we were able to isolate high-quality RNA (A260nm/A280nm >1.9 and A260nm/A230nm >2.0). Highest yield was produced with the Zeng and Yang modified protocol (384±46µg ARN/g fresh weight). Furthermore, electrophoretic analysis showed the integrity of isolated RNA and the absence of DNA. Another proof of the high quality of our purified RNA was the successful cDNA synthesis and amplification of a segment of the M. dubia actin 1 gene. We report three modified protocols for isolation total RNA from leaves of M. dubia. The modified protocols are easy, rapid, low in cost, and effective for high-quality and quantity total RNA isolation suitable for cDNA synthesis and polymerase chain reaction.

  5. Long-Term Production and Delivery of Human Growth Hormone In vivo

    NASA Astrophysics Data System (ADS)

    Heartlein, Michael W.; Roman, Victoria A.; Jiang, Ji-Lei; Sellers, Joan W.; Zuliani, Antoinette M.; Treco, Douglas A.; Selden, Richard F.

    1994-11-01

    The application of somatic cell gene therapy to large patient populations will require the development of safe and practical approaches to the generation and characterization of genetically manipulated cells. Transkaryotic implantation is a gene therapy system based on the production of clonal strains of engineered primary and secondary cells, using nonviral methods. We demonstrate here that, on implantation, these clonal cell strains stably and reproducibly deliver pharmacologic quantities of protein for the lifetime of the experimental animals.

  6. Contributions of photosynthetic and non-photosynthetic cell types to leaf respiration in Vicia faba L. and their responses to growth temperature.

    PubMed

    Long, Benedict M; Bahar, Nur H A; Atkin, Owen K

    2015-11-01

    In intact leaves, mitochondrial populations are highly heterogeneous among contrasting cell types; how such contrasting populations respond to sustained changes in the environment remains, however, unclear. Here, we examined respiratory rates, mitochondrial protein composition and response to growth temperature in photosynthetic (mesophyll) and non-photosynthetic (epidermal) cells from fully expanded leaves of warm-developed (WD) and cold-developed (CD) broad bean (Vicia faba L.). Rates of respiration were significantly higher in mesophyll cell protoplasts (MCPs) than epidermal cell protoplasts (ECPs), with both protoplast types exhibiting capacity for cytochrome and alternative oxidase activity. Compared with ECPs, MCPs contained greater relative quantities of porin, suggesting higher mitochondrial surface area in mesophyll cells. Nevertheless, the relative quantities of respiratory proteins (normalized to porin) were similar in MCPs and ECPs, suggesting that ECPs have lower numbers of mitochondria yet similar protein complement to MCP mitochondria (albeit with lower abundance serine hydroxymethyltransferase). Several mitochondrial proteins (both non-photorespiratory and photorespiratory) exhibited an increased abundance in response to cold in both protoplast types. Based on estimates of individual protoplast respiration rates, combined with leaf cell abundance data, epidermal cells make a small but significant (2%) contribution to overall leaf respiration which increases twofold in the cold. Taken together, our data highlight the heterogeneous nature of mitochondrial populations in leaves, both among contrasting cell types and in how those populations respond to growth temperature. © 2015 John Wiley & Sons Ltd.

  7. Leading and Manning Army 21, 1984 Summer Study

    DTIC Science & Technology

    1984-11-01

    to the four major issues treated in this report which were: Manning the mobilized forces, Recruiting and retention - Total Army, Impact of Quality of Life , and Quality and quantity of manpower for the future.

  8. 40 CFR 180.900 - Exemptions from the requirement of a tolerance.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From... granted when it appears that the total quantity of the pesticide chemical in or on all raw agricultural...

  9. 40 CFR 180.900 - Exemptions from the requirement of a tolerance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From... granted when it appears that the total quantity of the pesticide chemical in or on all raw agricultural...

  10. 7 CFR 948.132 - Issuance of exemption certificates.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... applicant, the location of his farm or ranch, the location, or locations, of all potatoes remaining to be shipped, the total quantity of potatoes which may be shipped under the certificate of exemption, and such...

  11. 7 CFR 948.132 - Issuance of exemption certificates.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... applicant, the location of his farm or ranch, the location, or locations, of all potatoes remaining to be shipped, the total quantity of potatoes which may be shipped under the certificate of exemption, and such...

  12. 7 CFR 948.132 - Issuance of exemption certificates.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... applicant, the location of his farm or ranch, the location, or locations, of all potatoes remaining to be shipped, the total quantity of potatoes which may be shipped under the certificate of exemption, and such...

  13. 7 CFR 948.132 - Issuance of exemption certificates.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... applicant, the location of his farm or ranch, the location, or locations, of all potatoes remaining to be shipped, the total quantity of potatoes which may be shipped under the certificate of exemption, and such...

  14. 7 CFR 948.132 - Issuance of exemption certificates.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... applicant, the location of his farm or ranch, the location, or locations, of all potatoes remaining to be shipped, the total quantity of potatoes which may be shipped under the certificate of exemption, and such...

  15. 27 CFR 19.599 - Bottling and packaging records.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... spirits bottled or packaged (if different from the proof recorded under paragraph (f) of this section); (k) Total quantity bottled, packaged, or otherwise disposed of in bulk; (l) Losses or gains of the distilled...

  16. 27 CFR 19.599 - Bottling and packaging records.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... spirits bottled or packaged (if different from the proof recorded under paragraph (f) of this section); (k) Total quantity bottled, packaged, or otherwise disposed of in bulk; (l) Losses or gains of the distilled...

  17. 27 CFR 19.599 - Bottling and packaging records.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... spirits bottled or packaged (if different from the proof recorded under paragraph (f) of this section); (k) Total quantity bottled, packaged, or otherwise disposed of in bulk; (l) Losses or gains of the distilled...

  18. 27 CFR 19.599 - Bottling and packaging records.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... proof recorded under paragraph (f) of this section); (k) Total quantity bottled, packaged, or otherwise disposed of in bulk; (l) Losses or gains of the distilled spirits product; and (m) If labeled as bottled in...

  19. Radiation Response of Cancer Stem-Like Cells From Established Human Cell Lines After Sorting for Surface Markers

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Al-Assar, Osama; Muschel, Ruth J.; Mantoni, Tine S.

    2009-11-15

    Purpose: A subpopulation of cancer stem-like cells (CSLC) is hypothesized to exist in different cancer cell lines and to mediate radioresistance in solid tumors. Methods and Materials: Cells were stained for CSLC markers and sorted (fluorescence-activated cell sorter/magnetic beads) to compare foci and radiosensitivity of phosphorylated histone H2AX at Ser 139 (gamma-H2AX) in sorted vs. unsorted populations in eight cell lines from different organs. CSLC properties were examined using anchorage-independent growth and levels of activated Notch1. Validation consisted of testing tumorigenicity and postirradiation enrichment of CSLC in xenograft tumors. Results: The quantity of CSLC was generally in good agreement withmore » primary tumors. CSLC from MDA-MB-231 (breast) and Panc-1 and PSN-1 (both pancreatic) cells had fewer residual gamma-H2AX foci than unsorted cells, pointing to radioresistance of CSLC. However, only MDA-MB-231 CSLC were more radioresistant than unsorted cells. Furthermore, MDA-MB-231 CSLC showed enhanced anchorage-independent growth and overexpression of activated Notch1 protein. The expression of cancer stem cell surface markers in the MDA-MB-231 xenograft model was increased after exposure to fractionated radiation. In contrast to PSN-1 cells, a growth advantage for MDA-MB-231 CSLC xenograft tumors was found compared to tumors arising from unsorted cells. Conclusions: CSLC subpopulations showed no general radioresistant phenotype, despite the quantities of CSLC subpopulations shown to correspond relatively well in other reports. Likewise, CSLC characteristics were found in some but not all of the tested cell lines. The reported problems in testing for CSLC in cell lines may be overcome by additional techniques, beyond sorting for markers.« less

  20. Functional antigen binding by the defective B cells of CBA/N mice.

    PubMed

    Snippe, H; Merchant, B; Lizzio, E F; Inman, J K

    1982-01-01

    CBA/N mice have an X-linked B cell defect which prevents them from responding to nonmitogenic thymic independent (TI-2) antigens such as dinitrophenylated DNP-Ficoll (1,2). The F1 male progeny of CBA/N female mice express the same defect. Spleen cell suspensions from such defective mice (CBA/N X C3H/HeN F1 males) could not respond to DNP-Ficoll following in vitro immunization and subsequent transfer into irradiated, syngeneic, F1 male recipients as expected. In contrast, normal CBA/N X C3H/HeN F1 female spleen cells could respond and effect a "rescue"; they mounted strong plaque-forming cell responses 7 days after in vitro exposure to DNP-Ficoll and subsequent transfer into irradiated F1 male recipients. Defective F1 male spleen cells, however, could bind significant quantities of 125I-DNP-Ficoll after in vitro exposure. Extensive washing of these spleen cells could not reverse this binding. Such DNP-Ficoll-exposed and washed F1 male spleen cells could, after transfer, aid normal untreated F1 female cells in their rescue function. The defective F1 male spleen cells could convey immunogenic quantities of DNP-Ficoll to the "rescuing" F1 female cells. Mitomycin treatment of F1 male cells did not interfere with their conveyor function. Goat anti-mouse mu serum impeded the passive antigen conveyor function of defective F1 male cells as did prior exposure to high concentrations of free DNP hapten. Our data support the view that the B cell defect of CBA/N X C3H/HeN F1 male mice does not relate to antigen binding, but rather to an inability to be effectively triggered by certain cell-bound polymeric antigens.

  1. [Sorption of microorganisms by fiber materials].

    PubMed

    Nikovskaia, G N; Gordienko, A S; Globa, L I

    1986-01-01

    Candida guilliermondii and Escherichia coli cells were adsorbed on glass and basalt fibres with a similar specific surface, but with a different charge. The quantity of adsorbed microorganisms did not depend on the type and charge of a fibre surface. However, cells were adsorbed faster and more firmly on positively charged and uncharged fibres than on negatively charged fibres.

  2. Management of food industry waste employing vermicomposting technology.

    PubMed

    Garg, V K; Suthar, S; Yadav, Anoop

    2012-12-01

    This paper reports the vermicomposting of food industry sludges (FIS) mixed with different organic wastes employing Eisenia fetida. A total of 10 vermicomposting units containing different wastes combinations were established. After 15 weeks significant increase in total nitrogen (N(total)) (60-214%), total available phosphorous (P(avail)) (35.8-69.6%), total sodium (Na(total)) (39-95%), and total potassium (K(total)) (43.7-74.1%), while decrease in pH (8.45-19.7%), total organic carbon (OC(total)) (28.4-36.1%) and C:N ratio (61.2-77.8%) was recorded. The results indicated that FIS may be converted into good quality manure by vermicomposting if spiked with other organic wastes in appropriate quantities. Copyright © 2011 Elsevier Ltd. All rights reserved.

  3. Tooth-meshing-harmonic static-transmission-error amplitudes of helical gears

    NASA Astrophysics Data System (ADS)

    Mark, William D.

    2018-01-01

    The static transmission errors of meshing gear pairs arise from deviations of loaded tooth working surfaces from equispaced perfect involute surfaces. Such deviations consist of tooth-pair elastic deformations and geometric deviations (modifications) of tooth working surfaces. To a very good approximation, the static-transmission-error tooth-meshing-harmonic amplitudes of helical gears are herein expressed by superposition of Fourier transforms of the quantities: (1) the combination of tooth-pair elastic deformations and geometric tooth-pair modifications and (2) fractional mesh-stiffness fluctuations, each quantity (1) and (2) expressed as a function of involute "roll distance." Normalization of the total roll-distance single-tooth contact span to unity allows tooth-meshing-harmonic amplitudes to be computed for different shapes of the above-described quantities (1) and (2). Tooth-meshing harmonics p = 1, 2, … are shown to occur at Fourier-transform harmonic values of Qp, p = 1, 2, …, where Q is the actual (total) contact ratio, thereby verifying its importance in minimizing transmission-error tooth-meshing-harmonic amplitudes. Two individual shapes and two series of shapes of the quantities (1) and (2) are chosen to illustrate a wide variety of shapes. In most cases representative of helical gears, tooth-meshing-harmonic values p = 1, 2, … are shown to occur in Fourier-transform harmonic regions governed by discontinuities arising from tooth-pair-contact initiation and termination, thereby showing the importance of minimizing such discontinuities. Plots and analytical expressions for all such Fourier transforms are presented, thereby illustrating the effects of various types of tooth-working-surface modifications and tooth-pair stiffnesses on transmission-error generation.

  4. Effectiveness of computer-assisted anesthetic delivery system (sta™) in dental implant surgery: a prospective study

    PubMed Central

    GRASSI, F.R.; RAPONE, B.; SCARANO CATANZARO, F.; CORSALINI, M.; KALEMAJ, Z.

    2017-01-01

    SUMMARY Objectives. This prospective cohort study aimed to investigate effectiveness of Computerized Local Anesthesia (CLA) on oral implantology through estimation of pain and discomfort and total quantity of injected anesthetic. Methods. Forty-five consecutive patients whose treatment plan included immediate or late dental implants were included in this study. The main inclusion criteria comprised: previous implant intervention under conventional anesthesia (CA) during the past 3 years and no previous treatment of pain relief. All patients reported on a 0–10 scale on previous experience with CA, and new experience with CLA. The same CLA system, namely Single Tooth Anesthesia (STA) was used for all patients with half of the quantity normally used for CA. Data on quantity of anesthetic and reported ratings were collected and described. Potential associations and determinant variables were analysed through correlation analysis and regression models. Results. Out of 45 patients, 27 received post-extractive implant surgery whereas the rest 18 implant surgery on healed sites. The reported pain from STA (mean 1.6, SD 0.7) showed important difference as compared to CA (7.9, SD 1.2; z=5.873; p<0.0001). The comfort perceived during the STA ranged from 7 to 10 (mean 9.5, SD 0.79). A second injection with half of the initial dose was necessary in three cases only. Conclusions. STA system proved to be effective during interventions of dental implantology, by markedly reducing patients’ pain and discomfort and the total quantity of necessary anesthetic. PMID:29682255

  5. Quality of diet plans for weight loss featured in women's magazines. A cross-sectional descriptive study.

    PubMed

    Martinighi, Maiara; Koga da Silva, Edina Mariko

    2017-01-01

    Brazil has the fifth largest population of obese individuals in the world. Women's magazines publish a large number of diet plans, and therefore the objective of this study was to assess the quality of these plans. Cross-sectional descriptive study. We included the Brazilian women's magazines of highest circulation published between January and June 2014 that advertised diets for weight loss on their covers. We extracted the quantities of macro and micronutrients from each of these diet plans and compared these quantities with the World Health Organization nutritional guidelines for adult women. We also checked the total energy quantities of these plans, and any recommendations about water intake and physical activity. We identified 136 potentially eligible magazine issues; 41 were excluded and 95 issues of 6 different magazines were included in the study. We found that 83.1 % of the plans had carbohydrate and fiber levels below the recommendations. On the other hand, the protein and saturated fatty acid levels were above the recommendations in 97.8% and 95.7% of the plans, respectively; 75.7% of the diets had inadequate calcium levels and 70.5% had low iron levels. Only 30 plans specified the total daily quantity of dietary energy and in 53.3% of these, the information was inconsistent with our estimates; 20% of the plans had no recommendations on daily water intake and 37.5% did not give recommendations regarding physical activity practices. The diet plans for weight loss featured in Brazilian women's magazines are of low quality.

  6. Program and charts for determining shock tube, and expansion tunnel flow quantities for real air

    NASA Technical Reports Server (NTRS)

    Miller, C. G., III; Wilder, S. E.

    1975-01-01

    A computer program in FORTRAN 4 language was written to determine shock tube, expansion tube, and expansion tunnel flow quantities for real-air test gas. This program permits, as input data, a number of possible combinations of flow quantities generally measured during a test. The versatility of the program is enhanced by the inclusion of such effects as a standing or totally reflected shock at the secondary diaphragm, thermochemical-equilibrium flow expansion and frozen flow expansion for the expansion tube and expansion tunnel, attenuation of the flow in traversing the acceleration section of the expansion tube, real air as the acceleration gas, and the effect of wall boundary layer on the acceleration section air flow. Charts which provide a rapid estimation of expansion tube performance prior to a test are included.

  7. System and method for conditioning a hardwood pulp liquid hydrolysate

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Waite, Darrell; Arnold, Richard; St. Pierre, James

    2015-06-30

    A system and method for hardwood pulp liquid hydrolysate conditioning includes a first evaporator receives a hardwood mix extract and outputting a quantity of vapor and extract. A hydrolysis unit receives the extract, hydrolyzes and outputs to a lignin separation device, which separates and recovers a quantity of lignin. A neutralization device receives extract from the lignin separation device and a neutralizing agent, producing a mixture of solid precipitate and a fifth extract. The solid precipitate is removed from the fifth extract. A second evaporator removes a quantity of acid from the fifth extract in a vapor form. This vapormore » may be recycled to improve total acid recovery or discarded. A desalination device receives the diluted extract, separates out some of the acid and salt and outputs a desalinated solution.« less

  8. A High Resolution Tampa Bay Hydrodynamic Model and its Application to Residence Time Estimation and Salt Balance Diagnosis

    NASA Astrophysics Data System (ADS)

    Zheng, L.; Weisberg, R. H.

    2016-02-01

    A 3D, numerical circulation model, with high resolution (20 m) at important mass conveyances (inlets and rivers etc.), is developed to estimate the bulk residence time and diagnose the salt balances and salt fluxes for Tampa Bay estuary. These analyses are justified via quantitative comparisons between the simulation and observations of sea level, velocity and salinity. The non-tidal circulation is the primary agent for the flushing of Tampa Bay. Tides alone have a minor effect. Exceptions pertain to within a tidal excursion from the bay mouth and regions with multiple inlets where different tide phases aid in flushing. The fully 3D salt flux divergences (SFD) and fluxes vary spatially throughout the estuary. On experimental duration (three month) average, the total advective SFD is balanced primarily by the vertical diffusive SFD, except near the bottom of the channel where the horizontal diffusive SFD is also important. Instantaneously, the local rate of salinity change is controlled primarily by the advective SFD, with a secondary contribution by the vertical diffusive SFD everywhere and the horizontal diffusive SFD near the channel bottom. After decomposing the advective salt fluxes and their divergences into mean quantity and tidal pumping, the horizontal and vertical advective SFDs by the mean quantities are large and counterbalance, with their sum being a small but significant residual. The horizontal and vertical advective SFDs by tidal pumping are relatively small (when compared with the mean quantities) and counterbalance; but, when summed, their residual is comparable in magnitude to that by the mean quantities. So whereas the salt fluxes by tidal pumping are secondary importance to the salt fluxes by the mean quantities, their total flux divergences are of comparable importance. The salt flux 3D components vary along the Tampa Bay axis, and these findings may be typical of coastal plain estuaries given their geometrical complexities.

  9. Detection of the quantity of kinesin and microgravity-sensitive kinesin genes in rat bone marrow stromal cells grown in a simulated microgravity environment

    NASA Astrophysics Data System (ADS)

    Ni, Chengzhi; Wang, Chunyan; Li, Yuan; Li, Yinghui; Dai, Zhongquan; Zhao, Dongming; Sun, Hongyi; Wu, Bin

    2011-06-01

    Kinesin and kinesin-like proteins (KLPs) constitute a superfamily of microtubule motor proteins found in all eukaryotic organisms. Members of the kinesin superfamily are known to play important roles in many fundamental cellular and developmental processes. To date, few published studies have reported on the effects of microgravity on kinesin expression. In this paper, we describe the expression pattern and microgravity-sensitive genes of kinesin in rat bone marrow stromal cells cultured in a ground-based rotating bioreactor. The quantity of kinesin under the clinorotation condition was examined by immunoblot analysis with anti-kinesin. Furthermore, the distribution of kinesin at various times during clinorotation was determined by dual immunostaining, using anti-kinesin monoclonal antibody or anti-β-tubulin monoclonal antibody. In terms of kinesin quantity, we found that the ratios of the amounts of clinorotated/stationary KLPs decreased from clinorotation day 5 to day 10, although it increased on days 2 and 3. Immunofluorescence analysis revealed that kinesin in the nucleus was the first to be affected by simulated microgravity, following the kinesin at the periphery that was affected at various times during clinorotation. Real-time RT-PCR analysis of kinesin mRNA expression was performed and led to the identification of 3 microgravity-sensitive kinesin genes: KIF9, KIFC1, and KIF21A. Our results suggest that kinesin has a distinct expression pattern, and the identification of microgravity-sensitive kinesin genes offers insight into fundamental cell biology.

  10. 48 CFR 49.202 - Profit.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ....202 Federal Acquisition Regulations System FEDERAL ACQUISITION REGULATION CONTRACT MANAGEMENT... with the total work required by the contract (engineering estimates of the percentage of completion... contractor, with particular regard to— (i) Attainment of quantity and quality production; (ii) Reduction of...

  11. Dioxin and Dioxin-Like Compounds Toxic Equivalency Information

    EPA Pesticide Factsheets

    EPA requires that, in addition to the total grams released for the entire dioxin and dioxin-like compounds category, TRI facilities must report the quantity for each individual member on a new Form R Schedule 1.

  12. Characterization of Jamaican Delonix regia and Cassia fistula Seed Extracts

    PubMed Central

    Reid, Raymond; Rattray, Vaughn; Williams, Ruth; Denny, Marcel

    2016-01-01

    Delonix regia and Cassia fistula seed extracts were evaluated for their antioxidant activity, total phenolics, ash, zinc and fatty acid content. Fourier Transform Infrared Spectroscopy (FTIR) was utilized to assess the chemical functionalities present within the seeds. Antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Trolox equivalent antioxidant capacity (TEAC) assays. Total phenolics were determined by the Folin-Ciocalteu assay. Lipid extracts were characterized by nuclear magnetic resonance spectroscopy and gas chromatography/mass spectrometry. Zinc concentration was determined by atomic absorption spectroscopy. Extracts from the seeds of C. fistula had a higher antioxidant activity, free radical scavenging activity, and phenolic content than D. regia. FTIR revealed that the seeds are a rich source of protein with small quantities of fat. C. fistula extracts contained a higher percentage of total fat than D. regia. Palmitic acid was identified as the predominant saturated fatty acid in both extracts. Oleic acid and linoleic acid were identified in smaller quantities. Seed extracts may be considered for use in food and nutraceutical applications. PMID:27034834

  13. Gonadotrophin-inhibitory hormone receptor expression in the chicken pituitary gland: potential influence of sexual maturation and ovarian steroids.

    PubMed

    Maddineni, S; Ocón-Grove, O M; Krzysik-Walker, S M; Hendricks, G L; Proudman, J A; Ramachandran, R

    2008-09-01

    Gonadotrophin-inhibitory hormone (GnIH), a hypothalamic RFamide, has been found to inhibit gonadotrophin secretion from the anterior pituitary gland originally in birds and, subsequently, in mammalian species. The gene encoding a transmembrane receptor for GnIH (GnIHR) was recently identified in the brain, pituitary gland and gonads of song bird, chicken and Japanese quail. The objectives of the present study are to characterise the expression of GnIHR mRNA and protein in the chicken pituitary gland, and to determine whether sexual maturation and gonadal steroids influence pituitary GnIHR mRNA abundance. GnIHR mRNA quantity was found to be significantly higher in diencephalon compared to either anterior pituitary gland or ovaries. GnIHR mRNA quantity was significantly higher in the pituitaries of sexually immature chickens relative to sexually mature chickens. Oestradiol or a combination of oestradiol and progesterone treatment caused a significant decrease in pituitary GnIHR mRNA quantity relative to vehicle controls. GnIHR-immunoreactive (ir) cells were identified in the chicken pituitary gland cephalic and caudal lobes. Furthermore, GnIHR-ir cells were found to be colocalised with luteinising hormone (LH)beta mRNA-, or follicle-stimulating hormone (FSH)beta mRNA-containing cells. GnIH treatment significantly decreased LH release from anterior pituitary gland slices collected from sexually immature, but not from sexually mature chickens. Taken together, GnIHR gene expression is possibly down regulated in response to a surge in circulating oestradiol and progesterone levels as the chicken undergoes sexual maturation to allow gonadotrophin secretion. Furthermore, GnIHR protein expressed in FSHbeta or LHbeta mRNA-containing cells is likely to mediate the inhibitory effect of GnIH on LH and FSH secretion.

  14. Amending greenroof soil with biochar to affect runoff water quantity and quality.

    PubMed

    Beck, Deborah A; Johnson, Gwynn R; Spolek, Graig A

    2011-01-01

    Numbers of greenroofs in urban areas continue to grow internationally; so designing greenroof soil to reduce the amount of nutrients in the stormwater runoff from these roofs is becoming essential. This study evaluated changes in extensive greenroof water discharge quality and quantity after adding biochar, a soil amendment promoted for its ability to retain nutrients in soils and increase soil fertility. Prototype greenroof trays with and without biochar were planted with sedum or ryegrass, with barren soil trays used as controls. The greenroof trays were subjected to two sequential 7.4cm/h rainfall events using a rain simulator. Runoff from the rain events was collected and evaluated. Trays containing 7% biochar showed increased water retention and significant decreases in discharge of total nitrogen, total phosphorus, nitrate, phosphate, and organic carbon. The addition of biochar to greenroof soil improves both runoff water quality and retention. Copyright © 2011 Elsevier Ltd. All rights reserved.

  15. Docosahexaenoic acid attenuates oxidative stress and protects human gingival fibroblasts against cytotoxicity induced by hydrogen peroxide and butyric acid.

    PubMed

    Zgorzynska, Emilia; Wierzbicka-Ferszt, Anita; Dziedzic, Barbara; Witusik-Perkowska, Monika; Zwolinska, Anna; Janas, Anna; Walczewska, Anna

    2015-01-01

    The oxidative burst of the host cells associated with bacterial pathogen infection contributes to the destruction of periodontal tissue. The present study investigates the effect of docosahexaenoic acid (DHA) on human gingival fibroblast (HGF) viability and ROS generation. The cell viability by MTT assay, ROS level using H2DCF-DA probe, and protein thiol content were measured in HGFs after 24h preincubation with different concentrations of DHA followed by treatment with H2O2. The cell death rate was determined by Annexin V/propidium iodide staining, and mitochondrial membrane potential (ΔΨm) was examined by MitoTracker Red probe in H2O2- and butyric acid-treated HGFs. The fatty acid composition of plasma membranes after incubation with DHA was determined by gas chromatography mass spectrometry. DHA preincubation in a dose-dependent manner increased the viability of HGFs exposed to H2O2 and decreased ROS generation compared to the control cells. In HGFs preincubated with 30μM DHA, the ΔΨm significantly increased in both H2O2- and butyric acid-treated cells. Moreover, incubation with DHA preserved the protein thiol level as effectively as N-acetylcysteine. Application of 50μM DHA increased the quantity of viable cells, decreased the number of necrotic cells after H2O2 treatment, and protected HGFs from apoptosis induced by butyric acid. DHA in the plasma membranes of these HGFs represented about 6% of the total amount of fatty acids. These results demonstrate that enrichment of HGFs with DHA reduces ROS generation and enhances the mitochondrial membrane potential protecting the fibroblasts against cytotoxic factors. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. 78 FR 56646 - Determination of Total Amounts of Fiscal Year 2014 WTO Tariff-Rate Quotas for Raw Cane Sugar and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-13

    ... Secretary Determination of Total Amounts of Fiscal Year 2014 WTO Tariff- Rate Quotas for Raw Cane Sugar and... the Fiscal Year (FY) 2014 (October 1, 2013-September 30, 2014) in-quota aggregate quantity of raw cane sugar at 1,117,195 metric tons raw value (MTRV). The Secretary also announces the establishment of the...

  17. 75 FR 47258 - Determination of Total Amounts of Fiscal Year 2011 Tariff-Rate Quotas for Raw Cane Sugar and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-05

    ... Determination of Total Amounts of Fiscal Year 2011 Tariff-Rate Quotas for Raw Cane Sugar and Certain Sugars...) 2011 in-quota aggregate quantity of the raw, as well as, refined and specialty sugar Tariff-Rate Quotas (TRQ) as required under the U.S. World Trade Organization (WTO) commitments. The FY 2011 raw cane sugar...

  18. 77 FR 55451 - Determination of Total Amounts of Fiscal Year 2013 Tariff-Rate Quotas for Raw Cane Sugar and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-10

    ... Secretary Determination of Total Amounts of Fiscal Year 2013 Tariff-Rate Quotas for Raw Cane Sugar and...) 2013 (October 1, 2012-September 30, 2013) in-quota aggregate quantity of the raw, as well as, refined and specialty sugar Tariff-Rate Quotas (TRQ). The FY 2013 raw cane sugar TRQ is established at 1,117...

  19. Development and Application of an Approach to Optimize Renewable Energy Systems in Afghanistan

    DTIC Science & Technology

    2012-06-01

    upon renewable energy sources for power production , the more desirable the system design. Total operations and maintenance cost has the third...Engineers (USACE) practices for implementing energy systems for ANSF infrastructure are limited to diesel generators, and, thus, preclude alternative...system attribute values: total O&M cost, renewable fraction, generator production , wind production , solar production , battery quantity, life cycle

  20. Quantity and quality of runoff from selected guttered and unguttered roadways in northeastern Ramsey County, Minnesota

    USGS Publications Warehouse

    Mitton, G.B.; Payne, G.A.

    1997-01-01

    Length of latent period was statistically compared to constituent concentration levels of total phosphorus, dissolved sulfate, and total zinc and there was a correlation. Constituent loads were not associated with latent period. No correlation was found between traffic volumes which ranged from 1,888 to 7,172 vehicles per day and constituent concentrations or loads for this study.

  1. Linear actuation using milligram quantities of CL-20 and TAGDNAT.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Snedigar, Shane; Salton, Jonathan Robert; Tappan, Alexander Smith

    2009-07-01

    There are numerous applications for small-scale actuation utilizing pyrotechnics and explosives. In certain applications, especially when multiple actuation strokes are needed, or actuator reuse is required, it is desirable to have all gaseous combustion products with no condensed residue in the actuator cylinder. Toward this goal, we have performed experiments on utilizing milligram quantities of high explosives to drive a millimeter-diameter actuator with a stroke of 30 mm. Calculations were performed to select proper material quantities to provide 0.5 J of actuation energy. This was performed utilizing the thermochemical code Cheetah to calculate the impetus for numerous propellants and tomore » select quantities based on estimated efficiencies of these propellants at small scales. Milligram quantities of propellants were loaded into a small-scale actuator and ignited with an ignition increment and hot wire ignition. Actuator combustion chamber pressure was monitored with a pressure transducer and actuator stroke was monitored using a laser displacement meter. Total actuation energy was determined by calculating the kinetic energy of reaction mass motion against gravity. Of the materials utilized, the best performance was obtained with a mixture of 2,4,6,8,10,12-hexanitro-2,4,6,8,10,12-hexaazaisowurtzitane (CL-20) and bis-triaminoguanidinium(3,3{prime}dinitroazotriazolate) (TAGDNAT).« less

  2. Effect of indirect non-thermal plasma on particle size distribution and composition of diesel engine particles

    NASA Astrophysics Data System (ADS)

    Linbo, GU; Yixi, CAI; Yunxi, SHI; Jing, WANG; Xiaoyu, PU; Jing, TIAN; Runlin, FAN

    2017-11-01

    To explore the effect of the gas source flow rate on the actual diesel exhaust particulate matter (PM), a test bench for diesel engine exhaust purification was constructed, using indirect non-thermal plasma technology. The effects of different gas source flow rates on the quantity concentration, composition, and apparent activation energy of PM were investigated, using an engine exhaust particle sizer and a thermo-gravimetric analyzer. The results show that when the gas source flow rate was large, not only the maximum peak quantity concentrations of particles had a large drop, but also the peak quantity concentrations shifted to smaller particle sizes from 100 nm to 80 nm. When the gas source flow rate was 10 L min-1, the total quantity concentration greatly decreased where the removal rate of particles was 79.2%, and the variation of the different mode particle proportion was obvious. Non-thermal plasma (NTP) improved the oxidation ability of volatile matter as well as that of solid carbon. However, the NTP gas source rate had little effects on oxidation activity of volatile matter, while it strongly influenced the oxidation activity of solid carbon. Considering the quantity concentration and oxidation activity of particles, a gas source flow rate of 10 L min-1 was more appropriate for the purification of particles.

  3. Master standard data quantity food production code. Macro elements for synthesizing production labor time.

    PubMed

    Matthews, M E; Waldvogel, C F; Mahaffey, M J; Zemel, P C

    1978-06-01

    Preparation procedures of standardized quantity formulas were analyzed for similarities and differences in production activities, and three entrée classifications were developed, based on these activities. Two formulas from each classification were selected, preparation procedures were divided into elements of production, and the MSD Quantity Food Production Code was applied. Macro elements not included in the existing Code were simulated, coded, assigned associated Time Measurement Units, and added to the MSD Quantity Food Production Code. Repeated occurrence of similar elements within production methods indicated that macro elements could be synthesized for use within one or more entrée classifications. Basic elements were grouped, simulated, and macro elements were derived. Macro elements were applied in the simulated production of 100 portions of each entrée formula. Total production time for each formula and average production time for each entrée classification were calculated. Application of macro elements indicated that this method of predetermining production time was feasible and could be adapted by quantity foodservice managers as a decision technique used to evaluate menu mix, production personnel schedules, and allocation of equipment usage. These macro elements could serve as a basis for further development and refinement of other macro elements which could be applied to a variety of menu item formulas.

  4. Novel method for the rapid isolation of RPE cells specifically for RNA extraction and analysis

    PubMed Central

    Wang, Cynthia Xin-Zhao; Zhang, Kaiyan; Aredo, Bogale; Lu, Hua; Ufret-Vincenty, Rafael L.

    2012-01-01

    RPE cells are involved in the pathogenesis of many retinal diseases. Accurate analysis of RPE gene expression profiles in different scenarios will increase our understanding of disease mechanisms. Our objective in this study was to develop an improved method for the isolation of RPE cells, specifically for RNA analysis. Mouse RPE cells were isolated using different techniques, including mechanical dissociation techniques and a new technique we refer to here as “Simultaneous RPE cell Isolation and RNA Stabilization” (SRIRS method). RNA was extracted from the RPE cells. An RNA bioanalyzer was used to determine the quantity and quality of RNA. qPCR was used to determine contamination with non-RPE-derived RNA. Several parameters with a potential impact on the isolation protocol were studied and optimized. A marked improvement in the quantity and quality of RPE-derived RNA was obtained with the SRIRS technique. We could get the RPE in direct contact with the RNA protecting agent within 1 minute of enucleation, and the RPE isolated within 11 minutes of enucleation. There was no significant contamination with vascular, choroidal or scleral-derived RNA. We have developed a fast, easy and reliable method for the isolation of RPE cells that leads to a high yield of RPE-derived RNA while preserving its quality. We believe this technique will be useful for future studies looking at gene expression profiles of RPE cells and their role in the pathophysiology of retinal diseases. PMID:22721721

  5. Development of blueberry liquor: influence of distillate, sweetener and fruit quantity.

    PubMed

    Caldeira, Ilda; Lopes, Daniel; Delgado, Teresa; Canas, Sara; Anjos, Ofélia

    2018-02-01

    In this work different formulations of blueberry liquor were tested and characterised based on their physico-chemical and sensory characteristics. Three factors were evaluated: the distillate used to produce the liquor (wine spirit or grape marc spirit); the sweetener (white sugar or honey) and the fruit quantity (two doses). For each liquor, pH, total acidity, dry soluble solids content, dry extract, alcoholic strength, reducing sugars, colour intensity, methanol content, acetaldehyde and fusel alcohols were determined. Sensory tests were carried out with a trained panel. The three factors studied significantly influenced the physico-chemical features of the liquors, being the quantity of fruit the most discriminating factor, except for the volatile compounds which were mainly influenced by the distillate. As regards the sensory analysis, it was found that the most appreciated liquor was that prepared with wine spirit, sugar and a lower dose of blueberry, and the less appreciated formulation was the one made with grape marc spirit, honey and a lower quantity of blueberry. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  6. Quantity Discrimination in Wolves (Canis lupus).

    PubMed

    Utrata, Ewelina; Virányi, Zsófia; Range, Friederike

    2012-01-01

    Quantity discrimination has been studied extensively in different non-human animal species. In the current study, we tested 11 hand-raised wolves (Canis lupus) in a two-way choice task. We placed a number of food items (one to four) sequentially into two opaque cans and asked the wolves to choose the larger amount. Moreover, we conducted two additional control conditions to rule out non-numerical properties of the presentation that the animals might have used to make the correct choice. Our results showed that wolves are able to make quantitative judgments at the group, but also at the individual level even when alternative strategies such as paying attention to the surface area or time and total amount are ruled out. In contrast to previous canine studies on dogs (Canis familiaris) and coyotes (Canis latrans), our wolves' performance did not improve with decreasing ratio, referred to as Weber's law. However, further studies using larger quantities than we used in the current set-up are still needed to determine whether and when wolves' quantity discrimination conforms to Weber's law.

  7. Quantity Discrimination in Wolves (Canis lupus)

    PubMed Central

    Utrata, Ewelina; Virányi, Zsófia; Range, Friederike

    2012-01-01

    Quantity discrimination has been studied extensively in different non-human animal species. In the current study, we tested 11 hand-raised wolves (Canis lupus) in a two-way choice task. We placed a number of food items (one to four) sequentially into two opaque cans and asked the wolves to choose the larger amount. Moreover, we conducted two additional control conditions to rule out non-numerical properties of the presentation that the animals might have used to make the correct choice. Our results showed that wolves are able to make quantitative judgments at the group, but also at the individual level even when alternative strategies such as paying attention to the surface area or time and total amount are ruled out. In contrast to previous canine studies on dogs (Canis familiaris) and coyotes (Canis latrans), our wolves’ performance did not improve with decreasing ratio, referred to as Weber’s law. However, further studies using larger quantities than we used in the current set-up are still needed to determine whether and when wolves’ quantity discrimination conforms to Weber’s law. PMID:23181044

  8. Bulky waste quantities and treatment methods in Denmark.

    PubMed

    Larsen, Anna W; Petersen, Claus; Christensen, Thomas H

    2012-02-01

    Bulky waste is a significant and increasing waste stream in Denmark. However, only little research has been done on its composition and treatment. In the present study, data about collection methods, waste quantities and treatment methods for bulky waste were obtained from two municipalities. In addition a sorting analysis was conducted on combustible waste, which is a major fraction of bulky waste in Denmark. The generation of bulky waste was found to be 150-250 kg capita(-1) year(-1), and 90% of the waste was collected at recycling centres; the rest through kerbside collection. Twelve main fractions were identified of which ten were recyclable and constituted 50-60% of the total quantity. The others were combustible waste for incineration (30-40%) and non-combustible waste for landfilling (10%). The largest fractions by mass were combustible waste, bricks and tile, concrete, non-combustible waste, wood, and metal scrap, which together made up more than 90% of the total waste amounts. The amount of combustible waste could be significantly reduced through better sorting. Many of the waste fractions consisted of composite products that underwent thorough separation before being recycled. The recyclable materials were in many cases exported to other countries which made it difficult to track their destination and further treatment.

  9. [Relationships between vegetation characteristics and soil properties at different restoration stages on slope land with purple soils in Hengyang of Hunan Province, South-central China].

    PubMed

    Yang, Ning; Zou, Dong-Sheng; Yang, Man-Yuan; Hu, Li-Zhen; Zou, Fang-Ping; Song, Guang-Tao; Lin, Zhong-Gui

    2013-01-01

    By using space series to replace time series, this paper studied the relationships between the vegetation characteristics and soil properties at different restoration stages on the slope land with purple soils in Hengyang of Hunnan Province South-central China. There existed obvious differences in the soil physical and chemical properties at different restoration stages. From grassplot, grass-shrub, shrub to shrub-arbor, the soil organic matter, total and available N, and moisture contents increased markedly, soil bulk density had an obvious decrease, soil total and available P contents changed little, and soil pH decreased gradually, but no significant differences were observed among different restoration stages. At different restoration stages, the biomass of plant community had effects on the quantity and composition of soil microbes. The quantities of soil bacteria and fungi had significant positive correlations with the aboveground biomass of plant community, but the quantity of soil actinomycetes had less correlation with plant community's aboveground biomass. At different restoration stages, the activities of soil urease, protease, alkaline phosphatase, invertase, cellulase, catalase, and polyphenol oxidase decreased with increasing soil layer, and had significant positive correlations with plant community's richness and aboveground biomass.

  10. Inventory simulation tools: Separating nuclide contributions to radiological quantities

    NASA Astrophysics Data System (ADS)

    Gilbert, Mark R.; Fleming, Michael; Sublet, Jean-Christophe

    2017-09-01

    The activation response of a material is a primary factor considered when evaluating its suitability for a nuclear application. Various radiological quantities, such as total (becquerel) activity, decay heat, and γ dose, can be readily predicted via inventory simulations, which numerically evolve in time the composition of a material under exposure to neutron irradiation. However, the resulting data sets can be very complex, often necessarily resulting in an over-simplification of the results - most commonly by just considering total response metrics. A number of different techniques for disseminating more completely the vast amount of data output from, in particular, the FISPACT-II inventory code system, including importance diagrams, nuclide maps, and primary knock-on atom (PKA) spectra, have been developed and used in scoping studies to produce database reports for the periodic table of elements. This paper introduces the latest addition to this arsenal - standardised and automated plotting of the time evolution in a radiological quantity for a given material separated by contributions from dominant radionuclides. Examples for relevant materials under predicted fusion reactor conditions, and for bench-marking studies against decay-heat measurements, demonstrate the usefulness and power of these radionuclide-separated activation plots. Note to the reader: the pdf file has been changed on September 22, 2017.

  11. [Analysis of academic impact of publications from National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention based on SCIE database in recent five years].

    PubMed

    Zhen, Li; Bin, Zheng

    2017-03-02

    To describe the academic impact of publications from National Institute of Parasitic Diseases (NIPD), Chinese Center for Disease Control and Prevention, so as to give the quantity evidence for scientific research decision making. The SCIE papers of NIPD published from 2011-2015 were searched and statistically analyzed. The number of published papers, citation frequencies, h-index, and funding resources were analyzed. The academic impact of the institute was assessed according to these data. A total of 361 papers were published by NIPD, and the quantity increased year by year. The majority type is original articles. The total citations were 1 641 times, the average citation per paper was 5.19 and h-index was 17. The majority of these papers were published in foreign professional periodicals, whose impact factors were between 1.194 and 6.751. The major resources of NIPD were from China, and NIPD also had good collaborations with institutions in US and Switzerland. In China and Asia, NIPD led the research in the field of parasitology and tropical medicine. The quantity and quality of annual published papers of NIPD are on the rise. However, NIPD lagged behind the leading institutions in the world.

  12. Murine maternal cell microchimerism: analysis using real-time PCR and in vivo imaging1

    PubMed Central

    Su, Eric C.; Johnson, Kirby L.; Tighiouart, Hocine; Bianchi, Diana W.

    2009-01-01

    In humans, maternal cells are present in the affected tissues of children with inflammatory myopathy, scleroderma, and neonatal lupus. It is unknown if maternal cell microchimerism (MCM) contributes to the pathology of disease. We sought to understand the factors that affect MCM to serve as a baseline for future mechanistic studies. Using a mouse model, we bred female mice transgenic for the luciferase (Luc) reporter gene to wild type (WT) males. The WT offspring were sacrificed at various postnatal ages. DNA was extracted from multiple organs and real-time PCR amplification was used to quantify Luc transgene as a marker for maternally derived cells. Sensitivity was 1 to 2 transgenic cells per 100,000 WT cells. MCM was noted in 85% of mice and 45% of tissues assayed. The average quantity of MCM was 158 maternal cells per 100,000 neonatal cells. The organs displaying the highest frequency and quantity of MCM were heart and lung (p<0.001). Postnatal age up to 21 days did not appear to affect levels of MCM (p=0.47), whereas increasing parity may increase levels of MCM. The data show that MCM is a common occurrence in healthy newborn mice, is present in their major organs, and that there are organ specific differences. This may represent differential migration of maternal cells, or varying receptivity of specific fetal organs to microchimerism. Pregnancy history appears to play a role in maternal cell trafficking. The role of MCM in pregnancy and disease pathogenesis remains to be elucidated. PMID:18256332

  13. On the bioavailability of trace metals in surface sediments: a combined geochemical and biological approach.

    PubMed

    Roosa, Stéphanie; Prygiel, Emilie; Lesven, Ludovic; Wattiez, Ruddy; Gillan, David; Ferrari, Benoît J D; Criquet, Justine; Billon, Gabriel

    2016-06-01

    The bioavailability of metals was estimated in three river sediments (Sensée, Scarpe, and Deûle Rivers) impacted by different levels of Cu, Cd, Pb, and Zn (Northern France). For that, a combination of geochemistry and biological responses (bacteria and chironomids) was used. The results obtained illustrate the complexity of the notion of "bioavailability." Indeed, geochemical indexes suggested a low toxicity, even in surface sediments with high concentrations of total metals and a predicted severe effect levels for the organisms. This was also suggested by the abundance of total bacteria as determined by DAPI counts, with high bacterial cell numbers even in contaminated areas. However, a fraction of metals may be bioavailable as it was shown for chironomid larvae which were able to accumulate an important quantity of metals in surface sediments within just a few days.We concluded that (1) the best approach to estimate bioavailability in the selected sediments is a combination of geochemical and biological approaches and that (2) the sediments in the Deûle and Scarpe Rivers are highly contaminated and may impact bacterial populations but also benthic invertebrates.

  14. Use of microbial activity measurements for monitoring RBC biofilms.

    PubMed

    Coello, M Dolores; Rodríguez-Barroso, M R; Aragón, C A; Quiroga, J M

    2010-10-01

    Fixed biomass technologies, such as rotating biological contactors or biodiscs, have been applied for wastewater depuration both in large and medium-small-sized agglomerations. Biofilm's composition and microorganism activity are essential parameters for the successful operation and control of these systems. Biofilm's thickness and total dry weight have been widely used for biofilm's characterization but, actually, are not sufficient to describe biofilm activity. In fact, biofilm's activity is not proportional to the quantity of fixed biomass, but increases with the thickness of biofilm up to a determined level called the "active thickness". Above this level, the diffusion of nutrients through the film becomes a limiting factor. A stable, thin, and active biofilm thus offers numerous advantages in water and wastewater treatment. Different parameters have been used to evaluate biofilm's activity. The specific oxygen uptake rate, INT-dehydrogenase activity, and the ratio active/total cells have been applied for that purpose. These methods are not only simple and rapid but also sensitive, precise, and representative. The results obtained confirm the potential of the microbial activity measurements studied for an accurate biofilm's characterization and biomass activity estimation in fundamental research and for the practical operation and control of fixed biomass depuration systems.

  15. THE MECHANISM OF THE INHIBITION OF HEMOLYSIS

    PubMed Central

    Ponder, Eric

    1945-01-01

    This paper contains a description of some of the inhibitory, and occasionally acceleratory, effects of sols of lecithins, cholesterol, and proteins in hemolytic systems containing simple lysins, together with investigations on the nature of the reactions by means of which the effects are brought about. The principal conclusions are: A. As regards sols of lecithins. 1. In lysin-inhibitor-cell systems, distearyl lecithin is an inhibitor of saponin and digitonin hemolysis, part of the effect being the result of a reaction with the components of the red cell surface and part being the result of a reaction with lysin in the bulk phase of the system. Lecithin ab ovo (Merck) is an accelerator of saponin hemolysis and either an accelerator or an inhibitor of digitonin hemolysis according to the initial concentration of lysin present in the system. Soybean lecithin is an inhibitor of both saponin and digitonin hemolysis, but both soybean lecithin and lecithin ab ovo contain also a hemolytic, or acceleratory, component. 2. The inhibitory effects depend on the order in which the components of the hemolytic system are mixed together. Distearyl lecithin is about 5 times more inhibitory in cell-inhibitor-lysin systems than in lysin-inhibitor-cell systems containing saponin, digitonin, or taurocholate. Lecithin ab ovo is more inhibitory in cell-inhibitor-lysin systems when the time of contact between cells and inhibitor is short, but when it is long, the hemolytic properties of the lecithin offset its inhibitory properties. A similar state of affairs is observed with soybean lecithin. 3. An increase in temperature decreases the inhibitory effect of distearyl lecithin in systems containing saponin or digitonin. B. As regards sols of cholesterol. 4. The quantity of lysin Δ apparently inhibited by a quantity Q of cholesterol sol is dependent on both the type of red cell and the number of red cells added to the system. 5. Δ is a non-linear function of Q and of c 1, the initial quantity of lysin present in the hemolytic system, Δ generally increasing as c 1 increases. 6. The inhibitory effect of cholesterol sols is essentially due to a reaction between the cholesterol and the lysin in the bulk phase of the system, modified by what appear to be redistribution effects which depend on the kind and number of red cells added to complete the hemolytic system. 7. The value of Δ depends on the temperature and on the length of time during which the cholesterol and the lysin remain in contact before the addition of the cells. 8. Distearyl lecithin considerably enhances the inhibitory effects of cholesterol sols. C. As regards the proteins. 9. Freshly prepared serum globulin is inhibitory in systems containing saponin, digitonin, taurocholate, and oleate, and the effect is due to reactions in the bulk phase of the system, modified by redistribution effects. 10. Serum albumin either accelerates or inhibits lysis by saponin, depending on the initial concentration of lysin, and the inhibition depends on such factors as the type of red cell used and the time of contact. In the case of sodium taurocholate, the inhibition has a very marked pH dependence. D. As regards plasma. 11. The way in which the inhibitory effect depends on the length of time during which inhibitor and lysin are in contact before the addition of the cells is not the same when plasma is used as an inhibitor as when a cholesterol sol is used as the inhibitor. The amount of cholesterol sol which is equal in inhibitory power to a given amount of plasma accordingly varies according to the length of the time of contact which is selected. 12. The inhibitory effect in systems containing saponin, plasma, and red cells can be shown to depend on the order in which the components are mixed, when the concentration of the plasma is small. 13. The question as to how much of the inhibitory power of plasma can be accounted for by the contained cholesterol (total or free) is one which can be answered only if the experimental conditions are defined with respect to initial concentration of lysin, time of contact, and several other variables. Very roughly, about 50 per cent of the total inhibition of plasma, or a little more, can be attributed to the cholesterol fraction. 14. Since the inhibitory effects of plasma are the result of reactions in the bulk phase of the system, complicated by redistributions among the phases, of reactions between some of its components and components of the red cell surface, and of enhancing effects of its components upon each other, it is not surprising that nothing better than an empirical expression should have been found to describe the inhibition quantitatively. PMID:19873439

  16. Estimates of ground-water pumpage from the Yakima River Basin aquifer system, Washington, 1960-2000

    USGS Publications Warehouse

    Vaccaro, J.J.; Sumioka, S.S.

    2006-01-01

    Ground-water pumpage in the Yakima River Basin, Washington, was estimated for eight categories of use for 1960-2000 as part of an investigation to assess groundwater availability in the basin. Methods used, pumpage estimates, reliability of the estimates, and a comparison with appropriated quantities are described. The eight categories of pumpage were public water supply, self-supplied domestic (exempt wells), irrigation, frost protection, livestock and dairy operations, industrial and commercial, fish and wildlife propagation, and ground-water claims. Pumpage estimates were based on methods that varied by the category and primarily represent pumpage for groundwater rights. Washington State Department of Ecology’s digital database has 2,874 active ground-water rights in the basin that can withdraw an annual quantity of about 529,231 acre-feet during dry years. Irrigation rights are for irrigation of about 129,570 acres. All but 220 of the rights were associated with well drillers’ logs, allowing for a spatial representation of the pumpage. Five-hundred and sixty of the irrigation rights were estimated to be standby/reserve rights. During this study, another 30 rights were identified that were not in the digital database. These rights can withdraw an annual quantity of about 20,969 acre-feet; about 6,700 acre-feet of these rights are near but outside the basin. In 1960, total annual pumpage in the basin, excluding standby/reserve pumpage, was about 115,776 acre-feet. By 2000, total annual pumpage was estimated to be 395,096 acre-feet, and excluding the standby/reserve rights, the total was 312,284 acre-feet. Irrigation accounts for about 60 percent of the pumpage, followed by public water supply at about 12 percent. The smallest category of pumpage was for livestock use with pumpage estimated to be 6,726 acre-feet. Total annual pumpage in 2000 was about 430 cubic feet per second, which is about 11 percent of the surface-water demand. Maximum pumpage is in July and August and during 2000, was about 100 cubic feet per second each month averaged over the Yakima River Basin aquifer system. During 2000, non-standby/reserve pumpage associated with ground-water rights was estimated to total 253,454 acre-feet, or about 198,290 acre-feet less than the appropriated quantity. The unused part of the appropriated value is about equivalent to the irrigation pumpage for primary rights.

  17. All-In-One: Advanced preparation of Human Parenchymal and Non-Parenchymal Liver Cells

    PubMed Central

    Werner, Melanie; Driftmann, Sabrina; Kleinehr, Kathrin; Kaiser, Gernot M.; Mathé, Zotlan; Treckmann, Juergen-Walter; Paul, Andreas; Skibbe, Kathrin; Timm, Joerg; Canbay, Ali; Gerken, Guido; Schlaak, Joerg F.; Broering, Ruth

    2015-01-01

    Background & Aims Liver cells are key players in innate immunity. Thus, studying primary isolated liver cells is necessary for determining their role in liver physiology and pathophysiology. In particular, the quantity and quality of isolated cells are crucial to their function. Our aim was to isolate a large quantity of high-quality human parenchymal and non-parenchymal cells from a single liver specimen. Methods Hepatocytes, Kupffer cells, liver sinusoidal endothelial cells, and stellate cells were isolated from liver tissues by collagenase perfusion in combination with low-speed centrifugation, density gradient centrifugation, and magnetic-activated cell sorting. The purity and functionality of cultured cell populations were controlled by determining their morphology, discriminative cell marker expression, and functional activity. Results Cell preparation yielded the following cell counts per gram of liver tissue: 2.0±0.4×107 hepatocytes, 1.8±0.5×106 Kupffer cells, 4.3±1.9×105 liver sinusoidal endothelial cells, and 3.2±0.5×105 stellate cells. Hepatocytes were identified by albumin (95.5±1.7%) and exhibited time-dependent activity of cytochrome P450 enzymes. Kupffer cells expressed CD68 (94.5±1.2%) and exhibited phagocytic activity, as determined with 1μm latex beads. Endothelial cells were CD146+ (97.8±1.1%) and exhibited efficient uptake of acetylated low-density lipoprotein. Hepatic stellate cells were identified by the expression of α-smooth muscle actin (97.1±1.5%). These cells further exhibited retinol (vitamin A)-mediated autofluorescence. Conclusions Our isolation procedure for primary parenchymal and non-parenchymal liver cells resulted in cell populations of high purity and quality, with retained physiological functionality in vitro. Thus, this system may provide a valuable tool for determining liver function and disease. PMID:26407160

  18. Scientific publications in nephrology and urology journals from Chinese authors in East Asia: a 10-year survey of the literature.

    PubMed

    Xu, Jing; Mao, Zhi-Guo; Kong, Mei; Hu, Liang-Hao; Ye, Chao-Yang; Xu, Cheng-Gang; Rong, Shu; Sun, Li-Jun; Wu, Jun; Dai, Bing; Chen, Dong-Ping; Zhu, Yu-Xian; Zhang, Yi-Xiang; Zhang, Yu-Qiang; Zhao, Xue-Zhi; Mei, Chang-Lin

    2011-04-08

    Diseases of the kidneys and genitourinary tract are common health problems that affect people of all ages and demographic backgrounds. In this study, we compared the quantity and quality of nephrological and urological articles published in international journals from the three major regions of China: the mainland (ML), Hong Kong (HK), and Taiwan (TW). Nephrological and urological articles originating from ML, TW, and HK that were published in 61 journals from 1999-2008 were retrieved from the PubMed database. We recorded the numbers of total articles, clinical trials, randomized controlled trials, case reports, impact factors (IF), citations, and articles published in the leading general-medicine journals. We used these data to compare the quantity and quality of publication output from the three regions. The total number of articles increased significantly from 1999 to 2008 in the three regions. The number of articles from ML has exceeded that from HK since 2004, and surpassed that from TW in 2008. Publications from TW had the highest accumulated IF, total citations of articles, and the most articles published in leading general-medicine journals. However, HK publications had the highest average IF. Although ML produced the largest quantity of articles, it exhibited the lowest quality among the three regions. The number of nephrological and urological publications originating from the three major regions of China increased significantly from 1999 to 2008. The annual number of publications by ML researchers exceeded those from TW and HK. However, the quality of articles from TW and HK was higher than that from ML.

  19. Electric Power Quarterly, October-December 1985. [Glossary

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Not Available

    1986-05-05

    The Electric Power Quarterly (EPQ) provides information on electric utilities at the plant level. The information concerns the following: cost, quantity, and quality of fossil fuel receipts; net generation; fuel consumption; and fuel stocks. The EPQ contains monthly data and quarterly totals for the reporting quarter. Data collected on Form EIA-759 regarding electric utilities' net generation, fuel consumption, and fuel stocks are presented on a plant-by-plant basis. In addition, quantity, cost, and quality of fossil fuel receipts collected on the Federal Energy Regulatory Commission (FERC) Form 423 are presented on a plant-by-plant basis.

  20. Electric Power Quarterly, January-March 1986

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Not Available

    1986-07-21

    The ''Electric Power Quarterly (EPQ)'' provides information on electric utilities at the plant level. The information concerns the following: cost, quantity, and quality of fossil fuel receipts; net generation; fuel consumption; and fuel stocks. The ''EPQ'' contains monthly data and quarterly totals for the reporting quarter. In this report, data collected on Form EIA-759 regarding electric utilities' net generation, fuel consumption, and fuel stocks are presented on a plant-by-plant basis. In addition, quantity, cost, and quality of fossil fuel receipts collected on the Federal Energy Regulatory Commission (FERC) Form 423 are presented on a plant-by-plant basis.

  1. Communication methods, systems, apparatus, and devices involving RF tag registration

    DOEpatents

    Burghard, Brion J [W. Richland, WA; Skorpik, James R [Kennewick, WA

    2008-04-22

    One technique of the present invention includes a number of Radio Frequency (RF) tags that each have a different identifier. Information is broadcast to the tags from an RF tag interrogator. This information corresponds to a maximum quantity of tag response time slots that are available. This maximum quantity may be less than the total number of tags. The tags each select one of the time slots as a function of the information and a random number provided by each respective tag. The different identifiers are transmitted to the interrogator from at least a subset of the RF tags.

  2. Photovoltaic Performance Enhancement of Silicon Solar Cells Based on Combined Ratios of Three Species of Europium-Doped Phosphors.

    PubMed

    Ho, Wen-Jeng; You, Bang-Jin; Liu, Jheng-Jie; Bai, Wen-Bin; Syu, Hong-Jhang; Lin, Ching-Fuh

    2018-05-18

    This paper presents a scheme for the enhancement of silicon solar cells in terms of luminescent emission band and photovoltaic performance. The proposed devices are coated with an luminescent down-shifting (LDS) layer comprising three species of europium (Eu)-doped phosphors mixed within a silicate film (SiO₂) using a spin-on film deposition. The three species of phosphor were mixed at ratios of 0.5:1:1.5, 1:1:1, or 1.5:1:0.5 in weight percentage (wt %). The total quantity of Eu-doped phosphors in the silicate solution was fixed at 3 wt %. The emission wavelengths of the Eu-doped phosphors were as follows: 518 nm (specie-A), 551 nm (specie-B), and 609 nm (specie-C). We examined the extended luminescent emission bands via photoluminescence measurements at room temperature. Closely matching the luminescent emission band to the high responsivity band of the silicon semiconductor resulted in good photovoltaic performance. Impressive improvements in efficiency were observed in all three samples: 0.5:1:1.5 (20.43%), 1:1:1 (19.67%), 1.5:1:0.5 (16.81%), compared to the control with a layer of pure SiO₂ (13.80%).

  3. Use of Dimethyl Pimelimidate with Microfluidic System for Nucleic Acids Extraction without Electricity.

    PubMed

    Jin, Choong Eun; Lee, Tae Yoon; Koo, Bonhan; Choi, Kyung-Chul; Chang, Suhwan; Park, Se Yoon; Kim, Ji Yeun; Kim, Sung-Han; Shin, Yong

    2017-07-18

    The isolation of nucleic acids in the lab on a chip is crucial to achieve the maximal effectiveness of point-of-care testing for detection in clinical applications. Here, we report on the use of a simple and versatile single-channel microfluidic platform that combines dimethyl pimelimidate (DMP) for nucleic acids (both RNA and DNA) extraction without electricity using a thin-film system. The system is based on the adaption of DMP into nonchaotropic-based nucleic acids and the capture of reagents into a low-cost thin-film platform for use as a microfluidic total analysis system, which can be utilized for sample processing in clinical diagnostics. Moreover, we assessed the use of the DMP system for the extraction of nucleic acids from various samples, including mammalian cells, bacterial cells, and viruses from human disease, and we also confirmed that the quality and quantity of the nucleic acids extracted were sufficient to allow for the robust detection of biomarkers and/or pathogens in downstream analysis. Furthermore, this DMP system does not require any instruments and electricity, and has improved time efficiency, portability, and affordability. Thus, we believe that the DMP system may change the paradigm of sample processing in clinical diagnostics.

  4. Protein-based composites and biomaterials

    USDA-ARS?s Scientific Manuscript database

    World industrialization has generated substantial quantities of petroleum-based plastics over many years, which are not biodegradable or compostable, and are permanently residing on land, in landfills, or in the oceans, as environmental pollution. Recently, total or partially degradable materials ha...

  5. 40 CFR 190.10 - Standards for normal operations.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ....10 Section 190.10 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) RADIATION PROTECTION PROGRAMS ENVIRONMENTAL RADIATION PROTECTION STANDARDS FOR NUCLEAR POWER OPERATIONS Environmental... operations and to radiation from these operations. (b) The total quantity of radioactive materials entering...

  6. 40 CFR 190.10 - Standards for normal operations.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ....10 Section 190.10 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) RADIATION PROTECTION PROGRAMS ENVIRONMENTAL RADIATION PROTECTION STANDARDS FOR NUCLEAR POWER OPERATIONS Environmental... operations and to radiation from these operations. (b) The total quantity of radioactive materials entering...

  7. Producing Quality Water for Industrial Use.

    ERIC Educational Resources Information Center

    Schaezler, Donald J.

    1978-01-01

    This article discusses the quality of water demanded by industrial plants and the techniques which are currently employed to achieve them. Both quality and quantity requirements are considered including total plant operation, physical and chemical operating controls, and systems monitoring. (CS)

  8. PHYSIOLOGY OF NITROGEN FIXATION BY BACILLUS POLYMYXA

    PubMed Central

    Grau, F. H.; Wilson, P. W.

    1962-01-01

    Grau, F. H. (University of Wisconsin, Madison) and P. W. Wilson. Physiology of nitrogen fixation by Bacillus polymyxa. J. Bacteriol. 83:490–496. 1962.—Of 17 strains of Bacillus polymyxa tested for fixation of molecular nitrogen, 15 fixed considerable quantities (30 to 150 μg N/ml). Two strains of the closely related B. macerans did not use N2, but possibly other members of this species may do so. Confirmation of fixation was obtained by showing incorporation of N15 into cell material. Both iron and molybdenum are specifically required for fixation; without the addition of these metals to the nitrogen-free medium, the growth rate and the total nitrogen fixed were reduced about 30 to 50%. No requirement for added molybdenum could be shown when ammonia was the nitrogen source, and the absence of iron caused only a slight decrease in growth. Washed-cell suspensions of B. polymyxa containing an active hydrogenase readily incorporated N15 into cell materials when provided with mannitol, glucose, or pyruvate but not when formate was the substrate. Hydrogen is a specific inhibitor of fixation, reducing both the rate and final amount of nitrogen fixed; it did not reduce growth on ammonia. Fixation was strictly anaerobic, 1% oxygen in the gas phase being sufficient to stop fixation. Arsenate is a powerful inhibitor of fixation of N2 by washed-cell suspensions of B. polymyxa, indicating that high-energy phosphate may be significant for this process. PMID:13901244

  9. Access to improved water and its relationship with diarrhoea in Kathmandu Valley, Nepal: a cross-sectional study

    PubMed Central

    Shrestha, Salina; Aihara, Yoko; Yoden, Kanako; Yamagata, Zentaro; Nishida, Kei; Kondo, Naoki

    2013-01-01

    Objective To assess the associations between diarrhoea and types of water sources, total quantity of water consumed and the quantity of improved water consumed in rapidly growing, highly populated urban areas in developing countries. Design Cross-sectional analysis using population-representative secondary data obtained from an interview survey conducted by the Asian Development Bank for the 2009 Kathmandu Valley Water Distribution, Sewerage and Urban Development Project. Setting Kathmandu Valley, Nepal. Participants 2282 households. Methods A structured questionnaire was used to collect information from households on the quantity and sources of water consumed; health, socioeconomic and demographic status of households; drinking water treatment practices and toilet facilities. Results Family members of 179 households (7.8%) reported having developed diarrhoea during the previous month. For households in which family members consumed less than 100 L of water per capita per day (L/c/d), which is the minimum quantity recommended by WHO, the risk of contracting diarrhoea doubled (1.56-fold to 2.92-fold). In households that used alternative water sources (such as wells, stone spouts and springs) in addition to improved water (provided by a water management authority), the likelihood of contracting diarrhoea was 1.81-fold higher (95% CI 1.00 to 3.29) than in those that used only improved water. However, access to an improved water source was not associated with a lower risk of developing diarrhoea if optimal quantities of water were not consumed (ie, <100 L/c/d). These results were independent of socioeconomic and demographic variables, daily drinking water treatment practices, toilet facilities and residential areas. Conclusions Providing access to a sufficient quantity of water—regardless of the source—may be more important in preventing diarrhoea than supplying a limited quantity of improved water. PMID:23811169

  10. Economic grand rounds: the price is right? Changes in the quantity of services used and prices paid in response to parity.

    PubMed

    Goldman, Howard H; Barry, Colleen L; Normand, Sharon-Lise T; Azzone, Vanessa; Busch, Alisa B; Huskamp, Haiden A

    2012-02-01

    The impact of parity coverage on the quantity of behavioral health services used by enrollees and on the prices of these services was examined in a set of Federal Employees Health Benefit (FEHB) Program plans. After parity implementation, the quantity of services used in the FEHB plans declined in five service categories, compared with plans that did not have parity coverage. The decline was significant for all service types except inpatient care. Because a previous study of the FEHB Program found that total spending on behavioral health services did not increase after parity implementation, it can be inferred that average prices must have increased over the period. The finding of a decline in service use and increase in prices provides an empirical window on what might be expected after implementation of the federal parity law and the parity requirement under the health care reform law.

  11. Investigation of grass carp by-products from a fish farm in Vojvodina

    NASA Astrophysics Data System (ADS)

    Okanović, Đ.; Tasić, T.; Kormanjoš, Š.; Ikonić, P.; Šojić, B.; Pelić, M.; Ristić, M.

    2017-09-01

    The quantity of by-products obtained during grass carp primary processing and chemical characteristics of internal organs were investigated. The total average weight of byproducts was 783.69 g (36.99%) in relation to live body weight which was cca 2118.5 g. The by-product contributing the largest quantity to total live body weight was the head with 458.22 g (21.63% of live body weight), followed by complete internal organs and tail and fins, with weights of 198.03 g or 9.35% and 57.93 g or 2.73%, respectively. The chemical composition of internal organs from the grass carp was mostly water (65.55%), following by crude fats and crude proteins (17.47% and 13.35%, respectively). The low collagen content (13.43% of total crude protein) indicates the high nutritional quality of the protein content from internal organs. Nitrogenous complexes from the internal organs were predominantly proteins. Digestible nitrogen was approximately equal to total nitrogen (89.38%), indicating that all proteins of the internal organs had high biological value. Based on the results obtained, it can be concluded that carp internal organs could be important sources of proteins and fats, and thus, could be used in Serbia as a raw material for feed and technical fat production.

  12. Analysis of tissue specific progenitor cell differentiation using FT-IR

    NASA Astrophysics Data System (ADS)

    Ishii, Katsunori; Kimura, Akinori; Kushibiki, Toshihiro; Awazu, Kunio

    2007-07-01

    Tissue specific progenitor cells and its differentiations have got a lot of attentions in regenerative medicine. The process of differentiations, the formation of tissues, has become better understood by the study using a lot of cell types progressively. These studies of cells and tissue dynamics at molecular levels are carried out through various approaches like histochemical methods, application of molecular biology and immunology. However, in case of using regenerative sources (cells, tissues and biomaterials etc.) clinically, they are measured and quality-controlled by non-contact and non-destructive methods from the view point of safety. Or the analysis with small quantities of materials could be possible if the quantities of materials are acceptable. A non-contact and non-destructive quality control method has been required. Recently, the use of Fourier Transform Infrared spectroscopy (FT-IR) has been used to monitor biochemical changes in cells, and has gained considerable importance. The changes in the cells and tissues, which are subtle and often not obvious in the histpathological studies, are shown to be well resolved using FT-IR. Moreover, although most techniques designed to detect one or a few changes, FT-IR is possible to identify the changes in the levels of various cellular biochemicals simultaneously under in vivo and in vitro conditions. The objective of this study is to establish the infrared spectroscopy of tissue specific progenitor cell differentiations as a quality control of cell sources for regenerative medicine. In the present study, as a basic study, we examine the adipose differentiation kinetics of preadipose cells (3T3-L1) and the osteoblast differentiation kinetics of mesenchymal stem cells (Kusa-A1) to analyze the infrared absorption spectra.

  13. Comparative analysis of the expression of c-Fos and interleukin-2 proteins in hypothalamus cells during various treatments.

    PubMed

    Barabanova, S V; Artyukhina, Z E; Ovchinnikova, K T; Abramova, T V; Kazakova, T B; Khavinson, V Kh; Malinin, V V; Korneva, E A

    2008-03-01

    The aim of the present work was to perform a combined analysis of the degree of activation of the anterior hypothalamus of the rat and expression of the interleukin-2 gene during treatments of different types: mild stress ("handling") and adaption to it, as well as intranasal administration of physiological saline and the peptides Vilon (Lys-Glu) and Epitalon (Ala-Glu-Asp-Gly). Changes in the numbers of c-Fos-and IL-2-positive cells in structures of the lateral area (LHA) and anterior (AHN), supraoptic (SON), and paraventricular (PVN) nuclei of the hypothalamus in Wistar rats. Ratios of the quantities of c-Fos-and IL-2-positive cells were determined in intact animals and after activation of brain cells initiated by different treatments; the influences of adaptation to handling on the nature of changes in the expression of these proteins was also studied. Combined analysis of the intensity of expression of these two proteins - c-Fos, a marker of neuron activation and a trans-factor for the IL-2 cytokine gene and other inducible genes, and IL-2 - in intact animals and after various treatments showed that the process of cell activation in most of the hypothalamic structures studied correlated with decreases in the quantity of IL-2-positive cells in these structures; different patterns of changes in the numbers of c-Fos-and IL-2-positive cells were seen in response to different treatments in conditions of stress and adaptation to it.

  14. 7 CFR 1212.70 - Reports.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... AND ORDERS; MISCELLANEOUS COMMODITIES), DEPARTMENT OF AGRICULTURE HONEY PACKERS AND IMPORTERS RESEARCH, PROMOTION, CONSUMER EDUCATION AND INDUSTRY INFORMATION ORDER Honey Packers and Importers Research, Promotion... necessary to perform its duties. (b) First handlers must report: (1) The total quantity of honey and honey...

  15. Contact integrity testing of stress-tested silicon terrestrial solar cells

    NASA Technical Reports Server (NTRS)

    Prince, J. L.; Lathrop, J. W.; Witter, G. W.

    1980-01-01

    A test procedure was developed and applied to terrestrial silicon solar cells in order to determine the effect of accelerated environmental and time-temperature aging on metal contact integrity. Quantities of cells of four different manufacturers were given the contact integrity test after being subjected to accelerated stress tests that included forward bias-temperature, thermal cycle and thermal shock, power cycle, and bias-temperature humidity tests at two temperature-humidity levels. Significant effects due to certain stress tests were found for some cell types. It is concluded that cells fabricated using plated nickel/solder metallization showed significantly more serious contact integrity degradation than silver-metallized cells.

  16. Identification of key factors regulating self-renewal and differentiation in EML hematopoietic precursor cells by RNA-sequencing analysis.

    PubMed

    Zong, Shan; Deng, Shuyun; Chen, Kenian; Wu, Jia Qian

    2014-11-11

    Hematopoietic stem cells (HSCs) are used clinically for transplantation treatment to rebuild a patient's hematopoietic system in many diseases such as leukemia and lymphoma. Elucidating the mechanisms controlling HSCs self-renewal and differentiation is important for application of HSCs for research and clinical uses. However, it is not possible to obtain large quantity of HSCs due to their inability to proliferate in vitro. To overcome this hurdle, we used a mouse bone marrow derived cell line, the EML (Erythroid, Myeloid, and Lymphocytic) cell line, as a model system for this study. RNA-sequencing (RNA-Seq) has been increasingly used to replace microarray for gene expression studies. We report here a detailed method of using RNA-Seq technology to investigate the potential key factors in regulation of EML cell self-renewal and differentiation. The protocol provided in this paper is divided into three parts. The first part explains how to culture EML cells and separate Lin-CD34+ and Lin-CD34- cells. The second part of the protocol offers detailed procedures for total RNA preparation and the subsequent library construction for high-throughput sequencing. The last part describes the method for RNA-Seq data analysis and explains how to use the data to identify differentially expressed transcription factors between Lin-CD34+ and Lin-CD34- cells. The most significantly differentially expressed transcription factors were identified to be the potential key regulators controlling EML cell self-renewal and differentiation. In the discussion section of this paper, we highlight the key steps for successful performance of this experiment. In summary, this paper offers a method of using RNA-Seq technology to identify potential regulators of self-renewal and differentiation in EML cells. The key factors identified are subjected to downstream functional analysis in vitro and in vivo.

  17. Identification of Key Factors Regulating Self-renewal and Differentiation in EML Hematopoietic Precursor Cells by RNA-sequencing Analysis

    PubMed Central

    Chen, Kenian; Wu, Jia Qian

    2014-01-01

    Hematopoietic stem cells (HSCs) are used clinically for transplantation treatment to rebuild a patient's hematopoietic system in many diseases such as leukemia and lymphoma. Elucidating the mechanisms controlling HSCs self-renewal and differentiation is important for application of HSCs for research and clinical uses. However, it is not possible to obtain large quantity of HSCs due to their inability to proliferate in vitro. To overcome this hurdle, we used a mouse bone marrow derived cell line, the EML (Erythroid, Myeloid, and Lymphocytic) cell line, as a model system for this study. RNA-sequencing (RNA-Seq) has been increasingly used to replace microarray for gene expression studies. We report here a detailed method of using RNA-Seq technology to investigate the potential key factors in regulation of EML cell self-renewal and differentiation. The protocol provided in this paper is divided into three parts. The first part explains how to culture EML cells and separate Lin-CD34+ and Lin-CD34- cells. The second part of the protocol offers detailed procedures for total RNA preparation and the subsequent library construction for high-throughput sequencing. The last part describes the method for RNA-Seq data analysis and explains how to use the data to identify differentially expressed transcription factors between Lin-CD34+ and Lin-CD34- cells. The most significantly differentially expressed transcription factors were identified to be the potential key regulators controlling EML cell self-renewal and differentiation. In the discussion section of this paper, we highlight the key steps for successful performance of this experiment. In summary, this paper offers a method of using RNA-Seq technology to identify potential regulators of self-renewal and differentiation in EML cells. The key factors identified are subjected to downstream functional analysis in vitro and in vivo. PMID:25407807

  18. Large number discrimination by mosquitofish.

    PubMed

    Agrillo, Christian; Piffer, Laura; Bisazza, Angelo

    2010-12-22

    Recent studies have demonstrated that fish display rudimentary numerical abilities similar to those observed in mammals and birds. The mechanisms underlying the discrimination of small quantities (<4) were recently investigated while, to date, no study has examined the discrimination of large numerosities in fish. Subjects were trained to discriminate between two sets of small geometric figures using social reinforcement. In the first experiment mosquitofish were required to discriminate 4 from 8 objects with or without experimental control of the continuous variables that co-vary with number (area, space, density, total luminance). Results showed that fish can use the sole numerical information to compare quantities but that they preferentially use cumulative surface area as a proxy of the number when this information is available. A second experiment investigated the influence of the total number of elements to discriminate large quantities. Fish proved to be able to discriminate up to 100 vs. 200 objects, without showing any significant decrease in accuracy compared with the 4 vs. 8 discrimination. The third experiment investigated the influence of the ratio between the numerosities. Performance was found to decrease when decreasing the numerical distance. Fish were able to discriminate numbers when ratios were 1:2 or 2:3 but not when the ratio was 3:4. The performance of a sample of undergraduate students, tested non-verbally using the same sets of stimuli, largely overlapped that of fish. Fish are able to use pure numerical information when discriminating between quantities larger than 4 units. As observed in human and non-human primates, the numerical system of fish appears to have virtually no upper limit while the numerical ratio has a clear effect on performance. These similarities further reinforce the view of a common origin of non-verbal numerical systems in all vertebrates.

  19. Tire-wear particles as a source of zinc to the environment

    USGS Publications Warehouse

    Councell, T.B.; Duckenfield, K.U.; Landa, E.R.; Callender, E.

    2004-01-01

    Tire-tread material has a zinc (Zn) content of about 1 wt %. The quantity of tread material lost to road surfaces by abrasion has not been well characterized. Two approaches were used to assess the magnitude of this nonpoint source of Zn in the U.S. for the period 1936-1999. In the first approach, tread-wear rates from the automotive engineering literature were used in conjunction with vehicle distance-driven data from the U.S. Department of Transportation to determine Zn releases. A second approach calculated this source term from the volume of tread lost during lifetime tire wear. These analyses showed that the quantity of Zn released by tire wear in the mid-1990s was of the same magnitude as that released from waste incineration. For 1999, the quantity of Zn released by tire wear in the U.S. is estimated to be 10 000-11 000 metric tons. A specific case study focused on Zn sources and sinks in an urban-suburban watershed (Lake Anne) in the Washington, DC, metropolitan area for a time period of the late 1990s. The atmospheric flux of total Zn (wet deposition) to the watershed was 2 ??g/cm2/yr. The flux of Zn to the watershed estimated from tire wear was 42 ??g/cm2/yr. The measured accumulation rate of total Zn in age-dated sediment cores from Lake Anne was 27 ??g/cm2/yr. These data suggest that tire-wear Zn inputs to urban-suburban watersheds can be significantly greater than atmospheric inputs, although the watershed appears to retain appreciable quantities of vehicular Zn inputs.

  20. Prescribed fires effects on physico-chemical properties and quantity of runoff and soil erosion in a Mediterranean forest

    NASA Astrophysics Data System (ADS)

    Esteban Lucas-Borja, Manuel; Plaza Alvaréz, Pedro Antonio; Sagra, Javier; Alfaro Sánchez, Raquel; Moya, Daniel; Ferrandiz Gotor, Pablo; De las Heras Ibañez, Jorge

    2017-04-01

    Wildfires have an important influence in forest ecosystems. Contrary to high severity fire, which may have negative impacts on the ecosystems, low severity induce small changes on soil properties. Thus and in order to reduce fire risk, low-severity prescribed fires have been widely used as a fuel reduction tool and silvicultural treatment in Mediterranean forest ecosystems. However, fire may alter microsite conditions and little is known about the impact of prescribed burning on the physico-chemical properties of runoff. In this study, we compared the effects of prescribed burning on physico-chemical properties and quantity of runoff and soil erosion during twelve months after a low severity prescribed fire applied in twelve 16 m2 plot (6 burned plots and 6 control plots used for comparison) set up in the Lezuza forest (Albacete, central-eastern Spain). Physico-chemical properties and quantity of runoff and soil losses were monitored after each rainfall event (five rainfall events in total). Also, different forest stand characteristics (slope, tree density, basal area and shrub/herbal cover) affecting each plot were measured. Results showed that forest stand characteristics were very similar in all used plots. Also, physico-chemical runoff properties were highly modified after the prescribed fire, increasing water pH, carbonates, bicarbonates, total dissolved solids and organic matter content dissolved in water. Electrical conductivity, calcium, sodium, chloride and magnesium were not affected by prescribed fire. Soil losses were highly related to precipitation intensity and tree interception. Tree intercepted the rainfall and significantly reduced soil losses and also runoff quantity. In conclusion and after the first six-month experiment, the influence of prescribed fires on physico-chemical runoff properties should be taken into account for developing proper prescribed burnings guidelines.

  1. Sleep Quantity and Quality during Acute Concussion: A Pilot Study

    PubMed Central

    Raikes, Adam C.; Schaefer, Sydney Y.

    2016-01-01

    Study Objectives: A number of subjective and objective studies provide compelling evidence of chronic post-concussion changes in sleep, yet very little is known about the acute effects of concussion on sleep quality and quantity. Therefore, the purpose of this prospective pilot study was to use actigraphy to examine the changes in sleep quality and quantity acutely following concussion at home rather than in a hospital or sleep laboratory. Methods: Seventeen young adults (7 with acute concussion, 10 controls) were recruited for this study. All participants completed two 5-day testing sessions separated by 30 days from intake (controls) or day of injury (concussion). Participants wore actigraphs and kept a sleep journal. Sleep parameter outcomes included nighttime total sleep time (nTST), 24-h total sleep time (TST), wake after sleep onset (WASO), and sleep efficiency (SE). The coefficient of variation (CV) for each sleep parameter was computed for each session. Results: nTST and TST CV was significantly greater in the concussion group. There is the additional indication that individuals with a concussion may require and obtain more sleep shortly after injury and subsequently have a shorter duration of sleep at 1 mo post-injury. This pattern was not seen in the measures of sleep quality (WASO, SE). Conclusions: Individuals with a concussion demonstrated increased nighttime sleep duration variability. This increase persisted at 1 mo post-injury and may be associated with previously documented self-reports of poor sleep quality lasting months and years after a concussion. Additionally, this increase may predispose individuals to numerous negative health outcomes if left untreated. Citation: Raikes AC, Schaefer SY. Sleep quantity and quality during acute concussion: a pilot study. SLEEP 2016;39(12):2141–2147. PMID:27748242

  2. LOX Tank Helium Removal for Propellant Scavenging

    NASA Technical Reports Server (NTRS)

    Chato, David J.

    2009-01-01

    System studies have shown a significant advantage to reusing the hydrogen and oxygen left in these tanks after landing on the Moon in fuel cells to generate power and water for surface systems. However in the current lander concepts, the helium used to pressurize the oxygen tank can substantially degrade fuel cell power and water output by covering the reacting surface with inert gas. This presentation documents an experimental investigation of methods to remove the helium pressurant while minimizing the amount of the oxygen lost. This investigation demonstrated that significant quantities of Helium (greater than 90% mole fraction) remain in the tank after draining. Although a single vent cycle reduced the helium quantity, large amounts of helium remained. Cyclic venting appeared to be more effective. Three vent cycles were sufficient to reduce the helium to small (less than 0.2%) quantities. Two vent cycles may be sufficient since once the tank has been brought up to pressure after the second vent cycle the helium concentration has been reduced to the less than 0.2% level. The re-pressurization process seemed to contribute to diluting helium. This is as expected since in order to raise the pressure liquid oxygen must be evaporated. Estimated liquid oxygen loss is on the order of 82 pounds (assuming the third vent cycle is not required).

  3. Piezoelectric axial flow microvalve

    DOEpatents

    Gemmen, Randall; Thornton, Jimmy; Vipperman, Jeffrey S.; Clark, William W.

    2007-01-09

    This invention is directed to a fuel cell operable with a quantity of fuel and a quantity of an oxidizer to produce electrical power, the fuel cell including a fuel cell body including a labyrinth system structured to permit the fuel and the oxidizer to flow therethrough; at least a first catalyst in fluid communication with the labyrinth; and at least a first microvalve operably disposed within at least a portion of the labyrinth. The microvalve utilizes a deflectable member operable upon the application of a voltage from a voltage source. The microvalve includes an elongated flow channel formed therein and extending substantially longitudinally between the first and second ends to permit substantially longitudinal flow of the fluid therethrough and between the first and second ends; and the deflectable member disposed on the valve body, the deflectable member including at least a first piezoelectric portion that is piezoelectrically operable to deflect the deflectable member between an open position and a closed position upon the application of a voltage, the deflectable member in the closed position being operable to resist the flow of the fluid through the flow channel.

  4. Accumulation and hyperaccumulation of copper in plants

    NASA Astrophysics Data System (ADS)

    Adam, V.; Trnkova, L.; Huska, D.; Babula, P.; Kizek, R.

    2009-04-01

    Copper is natural component of our environment. Flow of copper(II) ions in the environment depends on solubility of compounds containing this metal. Mobile ion coming from soil and rocks due to volcanic activity, rains and others are then distributed to water. Bio-availability of copper is substantially lower than its concentration in the aquatic environment. Copper present in the water reacts with other compounds and creates a complex, not available for organisms. The availability of copper varies depending on the environment, but moving around within the range from 5 to 25 % of total copper. Thus copper is stored in the sediments and the rest is transported to the seas and oceans. It is common knowledge that copper is essential element for most living organisms. For this reason this element is actively accumulated in the tissues. The total quantity of copper in soil ranges from 2 to 250 mg / kg, the average concentration is 30 mg / kg. Certain activities related to agriculture (the use of fungicides), possibly with the metallurgical industry and mining, tend to increase the total quantity of copper in the soil. This amount of copper in the soil is a problem particularly for agricultural production of food. The lack of copper causes a decrease in revenue and reduction in quality of production. In Europe, shows the low level of copper in total 18 million hectares of farmland. To remedy this adverse situation is the increasing use of copper fertilizers in agricultural soils. It is known that copper compounds are used in plant protection against various illnesses and pests. Mining of minerals is for the development of human society a key economic activity. An important site where the copper is mined in the Slovakia is nearby Smolníka. Due to long time mining in his area (more than 700 years) there are places with extremely high concentrations of various metals including copper. Besides copper, there are also detected iron, zinc and arsenic. Various plant species have adapted on such stress. The aim of this study is to investigate the behaviour of copper in plants and to assess its potential effect on the surrounding environment. To detect copper in biological samples electrochemical methods were employed particularly differential pulse voltammetry (DPV). Copper gave signals at 0.02 V measured by DPV. The obtained calibration dependence was linear (R2 = 0.995). Further, this method was utilized for determination of copper in real soil samples obtained from previously mentioned heavy-metal-polluted mining area. The content varied within range from tens to hundreds of mg of copper per kg of the soil. Moreover, we focused on investigation of copper influence on seedlings of Norway spruce. The seedlings were treated with copper (0, 0.1, 10 and 100 mM) for four weeks. We observed anatomical-morphological changes and other biochemical parameters in plants. We determined that seedlings synthesized more than 48 % protective thiols (glutathione and phytochelatins) compared to control ones. We investigated copper distribution in plant tissues by diphenylcarbazide staining. We found out that copper is highly accumulated in parenchymal stalk cells. In needles, change in auto-fluorescence of parenchymal cells of mesoderm similarly to endodermis cells. Besides, we analyzed samples of plants from the polluted area (spruce, pin, birch). The data obtained well correlated with previously mentioned. Acknowledgement The work on this experiment was supported by grant: INCHEMBIOL MSM0021622412.

  5. Effect of citronellol and the Chinese medical herb complex on cellular immunity of cancer patients receiving chemotherapy/radiotherapy.

    PubMed

    Zhuang, Shu-Ru; Chen, Su-Lin; Tsai, Jih-Hsin; Huang, Chi-Chou; Wu, Tzu-Chin; Liu, Wen-Shan; Tseng, Hsien-Chun; Lee, Hong-Sen; Huang, Min-Chang; Shane, Guang-Tzuu; Yang, Cheng-Hua; Shen, You-Cheng; Yan, Yeong-Yu; Wang, Chin-Kun

    2009-06-01

    Leukopenia and immunity impairment usually occur during cancer therapy. Citronellol, an oil soluble compound derived from the geranium, has anticancer and antiinflammatory properties, as well as promoting wound healing. Ganoderma lucidum, Codonopsis pilosula and Angelicae sinensis are traditional Chinese herbs, all of which have proven immunomodulatory functions in laboratory-based research. This randomized, double-blind, placebo-controlled study examined whether the Chinese medicinal herb complex (CCMH; a mixture of citronellol and extracts of G. lucidum, C. pilosula and A. sinensis) improves the immune cell counts of cancer patients receiving chemotherapy and/or radiotherapy. A total of 105 cancer patients receiving chemotherapy or radiotherapy were enrolled. The quantities of immune cells in the blood of the subjects were determined before and after 6 weeks of cancer treatment, with either CCMH or a placebo. CCMH significantly reduced the depletion of leukocytes (14.2% compared with 28.2%) and neutrophils (11.0% compared with 29.1%). Analysis of the lymphocyte phenotype revealed that the patients receiving the placebo had reduced CD4 lymphocytes and natural killer (NK) cells than the CCMH-treated patients. Treatment with CCMH for patients receiving chemotherapy and/or radiotherapy may improve their immune function, improving their ability to fight off the cancer, as well as any secondary infections that could compromise their treatment and their health. (c) 2009 John Wiley & Sons, Ltd.

  6. Sensitive and reliable detection of Kit point mutation Asp 816 to Val in pathological material

    PubMed Central

    Kähler, Christian; Didlaukat, Sabine; Feller, Alfred C; Merz, Hartmut

    2007-01-01

    Background Human mastocytosis is a heterogenous disorder which is linked to a gain-of-function mutation in the kinase domain of the receptor tyrosine kinase Kit. This D816V mutation leads to constitutive activation and phosphorylation of Kit with proliferative disorders of mast cells in the peripheral blood, skin, and spleen. Most PCR applications used so far are labour-intensive and are not adopted to daily routine in pathological laboratories. The method has to be robust and working on such different materials like archival formalin-fixed, paraffin-embedded tissue (FFPE) and blood samples. Such a method is introduced in this publication. Methods The Kit point mutation Asp 816 to Val is heterozygous which means a problem in detection by PCR because the wild-type allele is also amplified and the number of cells which bear the point mutation is in most of the cases low. Most PCR protocols use probes to block the wild-type allele during amplification with more or less satisfying result. This is why point-mutated forward primers were designed and tested for efficiency in amplification of the mutated allele. Results One primer combination (A) fits the most for the introduced PCR assay. It was able just to amplify the mutated allele with high specificity from different patient's materials (FFPE or blood) of varying quality and quantity. Moreover, the sensitivity for this assay was convincing because 10 ng of DNA which bears the point mutation could be detected in a total volume of 200 ng of DNA. Conclusion The PCR assay is able to deal with different materials (blood and FFPE) this means quality and quantity of DNA and can be used for high-througput screening because of its robustness. Moreover, the method is easy-to-use, not labour-intensive, and easy to realise in a standard laboratory. PMID:17900365

  7. Growth of Foot-and-Mouth Disease Virus in Dispersed Tissue Cells

    PubMed Central

    Patty, R. E.; Tozzini, F.; Seibold, H. R.; Callis, J. J.

    1962-01-01

    Methods are described for rapid and economical production of large quantities of foot-and-mouth disease virus in stationary cultures of trypsin-dispersed bovine kidney cells in a simple medium. Yields of between 107 and 108 plaque-forming units per milliliter were obtained from serum-free cultures containing approximately a million and a half viable trypsin-dispersed cells per milliliter. Some of the advantages and disadvantages of these methods of virus production are discussed. ImagesFig. 1Fig. 2Fig. 3 PMID:17649388

  8. Antigen-specific helper factors present in the supernatant of concanavalin A-induced spleen cell cultures.

    PubMed

    Kilburn, D G; Anaka, R

    1981-08-01

    The supernatants from cultures of concanavalin A-induced spleen cells contained both antigen-specific and nonspecific (Interleukin 2) helper factors (Hf). The antigen-specific factor could be isolated from the supernatant by adsorption onto and elution from antigen-Sepharose immunoadsorbents. Specific Hf was produced in cultures of either immune or nonimmune spleen cells although in the latter case the quantity of Hf was significantly less. The specific Hf did not manifest the thymocyte stimulatory property of 112.

  9. Post Remedial Action Report, Lansdowne Radioactive Residence Complex, Dismantlement/Removal Project. Volume 4. Radiological Oversight and Certification

    DTIC Science & Technology

    1990-06-01

    needed. [The quantity of radium] in the finished product was converted to the sulfate form and placed in platinum needles and cells made to the...of radon gas. The instrument incorporates a one-liter scintillation cell (a chamber lined with zinc sulfide phosphor sensitive to alpha particles) for...scintilla- tion cell at a flow rate of 1 L/min. The microprocessor is programmed to provide data output, in pCi/L. Normal data output is printed

  10. Enzyme Amplified Detection of Microbial Cell Wall Components

    NASA Technical Reports Server (NTRS)

    Wainwright, Norman R.

    2004-01-01

    This proposal is MBL's portion of NASA's Johnson Space Center's Astrobiology Center led by Principal Investigator, Dr. David McKay, entitled: 'Institute for the Study of Biomarkers in Astromaterials.' Dr. Norman Wainwright is the principal investigator at MBL and is responsible for developing methods to detect trace quantities of microbial cell wall chemicals using the enzyme amplification system of Limulus polyphemus and other related methods.

  11. Laser capture microdissection of embryonic cells and preparation of RNA for microarray assays.

    PubMed

    Redmond, Latasha C; Pang, Christopher J; Dumur, Catherine; Haar, Jack L; Lloyd, Joyce A

    2014-01-01

    In order to compare the global gene expression profiles of different embryonic cell types, it is first necessary to isolate the specific cells of interest. The purpose of this chapter is to provide a step-by-step protocol to perform laser capture microdissection (LCM) on embryo samples and obtain sufficient amounts of high-quality RNA for microarray hybridizations. Using the LCM/microarray strategy on mouse embryo samples has some challenges, because the cells of interest are available in limited quantities. The first step in the protocol is to obtain embryonic tissue, and immediately cryoprotect and freeze it in a cryomold containing Optimal Cutting Temperature freezing media (Sakura Finetek), using a dry ice-isopentane bath. The tissue is then cryosectioned, and the microscope slides are processed to fix, stain, and dehydrate the cells. LCM is employed to isolate specific cell types from the slides, identified under the microscope by virtue of their morphology. Detailed protocols are provided for using the currently available ArcturusXT LCM instrument and CapSure(®) LCM Caps, to which the selected cells adhere upon laser capture. To maintain RNA integrity, upon removing a slide from the final processing step, or attaching the first cells on the LCM cap, LCM is completed within 20 min. The cells are then immediately recovered from the LCM cap using a denaturing solution that stabilizes RNA integrity. RNA is prepared using standard methods, modified for working with small samples. To ensure the validity of the microarray data, the quality of the RNA is assessed using the Agilent bioanalyzer. Only RNA that is of sufficient integrity and quantity is used to perform microarray assays. This chapter provides guidance regarding troubleshooting and optimization to obtain high-quality RNA from cells of limited availability, obtained from embryo samples by LCM.

  12. Generation of human secondary cardiospheres as a potent cell processing strategy for cell-based cardiac repair.

    PubMed

    Cho, Hyun-Jai; Lee, Ho-Jae; Chung, Yeon-Ju; Kim, Ju-Young; Cho, Hyun-Ju; Yang, Han-Mo; Kwon, Yoo-Wook; Lee, Hae-Young; Oh, Byung-Hee; Park, Young-Bae; Kim, Hyo-Soo

    2013-01-01

    Cell therapy is a promising approach for repairing damaged heart. However, there are large rooms to be improved in therapeutic efficacy. We cultured a small quantity (5-10 mg) of heart biopsy tissues from 16 patients who received heart transplantation. We produced primary and secondary cardiospheres (CSs) using repeated three-dimensional culture strategy and characterized the cells. Approximately 5000 secondary CSs were acquired after 45 days. Genetic analysis confirmed that the progenitor cells in the secondary CSs originated from the innate heart, but not from extra-cardiac organs. The expressions of Oct4 and Nanog were significantly induced in secondary CSs compared with adherent cells derived from primary CSs. Those expressions in secondary CSs were higher in a cytokine-deprived medium than in a cytokine-supplemented one, suggesting that formation of the three-dimensional structure was important to enhance stemness whereas supplementation with various cytokines was not essential. Signal blocking experiments showed that the ERK and VEGF pathways are indispensable for sphere formation. To optimize cell processing, we compared four different methods of generating spheres. Method based on the hanging-drop or AggreWell™ was superior to that based on the poly-d-lysine-coated dish or Petri dish with respect to homogeneity of the product, cellular potency and overall simplicity of the process. When transplanted into the ischemic myocardium of immunocompromised mice, human secondary CSs differentiated into cardiomyocytes and endothelial cells. These results demonstrate that generation of secondary CSs from a small quantity of adult human cardiac tissue is a feasible and effective cell processing strategy to improve the therapeutic efficacy of cell therapy. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. Laser Capture Microdissection of Embryonic Cells and Preparation of RNA for Microarray Assays

    PubMed Central

    Redmond, Latasha C.; Pang, Christopher J.; Dumur, Catherine; Haar, Jack L.; Lloyd, Joyce A.

    2014-01-01

    In order to compare the global gene expression profiles of different embryonic cell types, it is first necessary to isolate the specific cells of interest. The purpose of this chapter is to provide a step-by-step protocol to perform laser capture microdissection (LCM) on embryo samples and obtain sufficient amounts of high-quality RNA for microarray hybridizations. Using the LCM/microarray strategy on mouse embryo samples has some challenges, because the cells of interest are available in limited quantities. The first step in the protocol is to obtain embryonic tissue, and immediately cryoprotect and freeze it in a cryomold containing Optimal Cutting Temperature freezing media (Sakura Finetek), using a dry ice–isopentane bath. The tissue is then cryosectioned, and the microscope slides are processed to fix, stain, and dehydrate the cells. LCM is employed to isolate specific cell types from the slides, identified under the microscope by virtue of their morphology. Detailed protocols are provided for using the currently available ArcturusXT LCM instrument and CapSure® LCM Caps, to which the selected cells adhere upon laser capture. To maintain RNA integrity, upon removing a slide from the final processing step, or attaching the first cells on the LCM cap, LCM is completed within 20 min. The cells are then immediately recovered from the LCM cap using a denaturing solution that stabilizes RNA integrity. RNA is prepared using standard methods, modified for working with small samples. To ensure the validity of the microarray data, the quality of the RNA is assessed using the Agilent bioanalyzer. Only RNA that is of sufficient integrity and quantity is used to perform microarray assays. This chapter provides guidance regarding troubleshooting and optimization to obtain high-quality RNA from cells of limited availability, obtained from embryo samples by LCM. PMID:24318813

  14. Quantifying the Labeling and the Levels of Plant Cell Wall Precursors Using Ion Chromatography Tandem Mass Spectrometry1[W][OA

    PubMed Central

    Alonso, Ana P.; Piasecki, Rebecca J.; Wang, Yan; LaClair, Russell W.; Shachar-Hill, Yair

    2010-01-01

    The biosynthesis of cell wall polymers involves enormous fluxes through central metabolism that are not fully delineated and whose regulation is poorly understood. We have established and validated a liquid chromatography tandem mass spectrometry method using multiple reaction monitoring mode to separate and quantify the levels of plant cell wall precursors. Target analytes were identified by their parent/daughter ions and retention times. The method allows the quantification of precursors at low picomole quantities with linear responses up to the nanomole quantity range. When applying the technique to Arabidopsis (Arabidopsis thaliana) T87 cell cultures, 16 hexose-phosphates (hexose-Ps) and nucleotide-sugars (NDP-sugars) involved in cell wall biosynthesis were separately quantified. Using hexose-P and NDP-sugar standards, we have shown that hot water extraction allows good recovery of the target metabolites (over 86%). This method is applicable to quantifying the levels of hexose-Ps and NDP-sugars in different plant tissues, such as Arabidopsis T87 cells in culture and fenugreek (Trigonella foenum-graecum) endosperm tissue, showing higher levels of galacto-mannan precursors in fenugreek endosperm. In Arabidopsis cells incubated with [U-13CFru]sucrose, the method was used to track the labeling pattern in cell wall precursors. As the fragmentation of hexose-Ps and NDP-sugars results in high yields of [PO3]−/or [H2PO4]− ions, mass isotopomers can be quantified directly from the intensity of selected tandem mass spectrometry transitions. The ability to directly measure 13C labeling in cell wall precursors makes possible metabolic flux analysis of cell wall biosynthesis based on dynamic labeling experiments. PMID:20442274

  15. Porous Nanogold/Polyurethane Scaffolds with Improved Antibiofilm, Mechanical, and Thermal Properties and with Reduced Effects on Cell Viability: A Suitable Material for Soft Tissue Applications.

    PubMed

    Tamayo, L; Acuña, D; Riveros, A L; Kogan, M J; Azócar, M I; Páez, M; Leal, M; Urzúa, M; Cerda, E

    2018-04-25

    The use of implants carries on a series of problems, among them infections, poor biocompatibility, high levels of cytotoxicity, and significant mechanical differences between implants and host organs that promote stress shielding effects. These problems indicate that the materials used to make implants must meet essential requirements and high standards for implantations to be successful. In this work, we present the synthesis, characterization and evaluation of the antibiofilm, mechanical, and thermal properties, and cytotoxic effect of a nanocomposite-based scaffold on polyurethane (PU) and gold nanoparticles (AuNPs) for soft tissue applications. The effect of the quantity of AuNPs on the antibacterial activity of nanocomposite scaffolds was evaluated against Staphylococcus epidermidis and Klebsiella spp., with a resulting 99.99% inhibition of both bacteria using a small quantity of nanoparticles. Cytotoxicity was evaluated with the T10 1/2 test against fibroblast cells. The results demonstrated that porous nanogold/PU scaffolds have no toxic effects on fibroblast cells to the 5 day exposition. With respect to mechanical properties, stress-strain curves showed that the compressive modulus and yield strength of PU scaffolds were significantly enhanced by AuNPs (by at least 10 times). This is due to changes in the arrangement of hard segments of PU, which increase the stiffness of the polymer. Thermogravimetric analysis showed that the degradation onset temperature rises with an increase in the quantity of AuNPs. These properties and characteristics demonstrate that porous nanogold/PU scaffolds are suitable material for use in soft tissue implants.

  16. RNA isolation from mammalian cells using porous polymer monoliths: an approach for high-throughput automation.

    PubMed

    Chatterjee, Anirban; Mirer, Paul L; Zaldivar Santamaria, Elvira; Klapperich, Catherine; Sharon, Andre; Sauer-Budge, Alexis F

    2010-06-01

    The life science and healthcare communities have been redefining the importance of ribonucleic acid (RNA) through the study of small molecule RNA (in RNAi/siRNA technologies), micro RNA (in cancer research and stem cell research), and mRNA (gene expression analysis for biologic drug targets). Research in this field increasingly requires efficient and high-throughput isolation techniques for RNA. Currently, several commercial kits are available for isolating RNA from cells. Although the quality and quantity of RNA yielded from these kits is sufficiently good for many purposes, limitations exist in terms of extraction efficiency from small cell populations and the ability to automate the extraction process. Traditionally, automating a process decreases the cost and personnel time while simultaneously increasing the throughput and reproducibility. As the RNA field matures, new methods for automating its extraction, especially from low cell numbers and in high throughput, are needed to achieve these improvements. The technology presented in this article is a step toward this goal. The method is based on a solid-phase extraction technology using a porous polymer monolith (PPM). A novel cell lysis approach and a larger binding surface throughout the PPM extraction column ensure a high yield from small starting samples, increasing sensitivity and reducing indirect costs in cell culture and sample storage. The method ensures a fast and simple procedure for RNA isolation from eukaryotic cells, with a high yield both in terms of quality and quantity. The technique is amenable to automation and streamlined workflow integration, with possible miniaturization of the sample handling process making it suitable for high-throughput applications.

  17. The role of prenatal exposures on body fat patterns at 7 years: Intrauterine programming or birthweight effects?

    PubMed

    Santos, S; Severo, M; Gaillard, R; Santos, A C; Barros, H; Oliveira, A

    2016-11-01

    It remains unknown whether the effects of prenatal exposures on child's adiposity reflect entirely intrauterine programming. We aimed to assess the effects of maternal gestational weight gain, diabetes and smoking on the child's body fat patterns, disentangling the direct (through intrauterine programming) and indirect (through birthweight) effects. We included 4747 singleton 7-year-old children from the Generation XXI birth cohort (Porto, Portugal). At birth, maternal and newborn's characteristics were obtained. Anthropometrics were measured at age 7 years and body fat patterns were identified by principal component analysis. Path analysis was used to quantify direct, indirect and total effects of gestational weight gain, diabetes and smoking on body fat patterns. Pattern 1 was characterized by strong factor loadings with body mass index, fat mass index and waist-to-height ratio (fat quantity) and pattern 2 with waist-to-hip ratio, waist-to-thigh ratio, and waist-to-weight ratio (fat distribution). The positive total effect of maternal gestational weight gain and diabetes on the child's fat quantity was mainly through a direct pathway, responsible for 91.7% and 83.7% of total effects, respectively (β = 0.022; 95% Confidence Interval (CI): 0.017, 0.027; β = 0.041; 95% CI: -0.011, 0.093). No effects on fat distribution were found. Maternal prenatal smoking had a positive direct effect on patterns 1 and 2, explaining 94.9% and 76.1% of total effects, respectively. The effects of maternal gestational weight gain, diabetes and smoking on a child's fat quantity seem to be mainly through intrauterine programming. Maternal smoking also showed a positive direct effect on child's fat distribution. Copyright © 2016 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.

  18. Inorganic salt mixtures as electrolyte media in fuel cells

    NASA Technical Reports Server (NTRS)

    Angell, Charles Austen (Inventor); Francis-Gervasio, Dominic (Inventor); Belieres, Jean-Philippe (Inventor)

    2012-01-01

    Fuel cell designs and techniques for converting chemical energy into electrical energy uses a fuel cell are disclosed. The designs and techniques include an anode to receive fuel, a cathode to receive oxygen, and an electrolyte chamber in the fuel cell, including an electrolyte medium, where the electrolyte medium includes an inorganic salt mixture in the fuel cell. The salt mixture includes pre-determined quantities of at least two salts chosen from a group consisting of ammonium trifluoromethanesulfonate, ammonium trifluoroacetate, and ammonium nitrate, to conduct charge from the anode to the cathode. The fuel cell includes an electrical circuit operatively coupled to the fuel cell to transport electrons from the cathode.

  19. Recombinant glucose uptake system

    DOEpatents

    Ingrahm, Lonnie O.; Snoep, Jacob L.; Arfman, Nico

    1997-01-01

    Recombinant organisms are disclosed that contain a pathway for glucose uptake other than the pathway normally utilized by the host cell. In particular, the host cell is one in which glucose transport into the cell normally is coupled to PEP production. This host cell is transformed so that it uses an alternative pathway for glucose transport that is not coupled to PEP production. In a preferred embodiment, the host cell is a bacterium other than Z. mobilis that has been transformed to contain the glf and glk genes of Z. mobilis. By uncoupling glucose transport into the cell from PEP utilization, more PEP is produced for synthesis of products of commercial importance from a given quantity of biomass supplied to the host cells.

  20. [Quantity and function of T follicular helper cells in the bone marrow of patients with immune thrombocytopenia].

    PubMed

    Zhang, Yang; Qu, Wen; Ruan, Er-Bao; Fu, Rong; Wang, Guo-Jin; Liu, Hong; Wang, Xiao-Ming; Wu, Yu-Hong; Song, Jia; Xing, Li-Min; Guan, Jing; Li, Li-Juan; Wang, Hua-Quan; Shao, Zong-Hong

    2014-06-01

    This study was purposed to detect the quantity and function of bone marrow (BM) T follicular helper (Tfh) cells of patients with immune thrombocytopenia, and to explore the role of Tfh cells in the pathogenesis of ITP. Twenty-one newly diagnosed ITP patients, twenty ITP patients in recovery stage and eighteen normal controls were enrolled in this study. The percentages of Tfh cells, Tfh-related molecules ICOS, CD40L, IL-21 in BM were detected by flow cytometry (FCM), and the mRNA expression of BCL-6 in BMMNC was determined by semi-quantitive RT-PCR. Correlation of Tfh cell level with the disease severity of ITP patients was analysed. The results showed that the ratio of CD4(+)CXCR5(+)/CD4(+) cells in newly diagnosed ITP patients [(5.532 ± 2.599)%] was significantly higher than that in ITP patients with recovery stage [(4.064 ± 2.026)%] and controls [(4.048 ± 1.413)%] (P < 0.05). The ratio of CD4(+)CXCR5(+)ICOS(+)/CD4(+) CXCR5(+) cells in newly diagnosed ITP patients [(14.586 ± 8.561)%] was higher than that in recovery stage ITP patients [(12.884 ± 10.161)%] and controls [(7.487 ± 5.176)%]. The differences be-tween newly diagnosed ITP patients and controls were statistically significant (P < 0.05). The ratio of CD4(+)CXCR5(+) CD40L(+)/CD4(+) CXCR5(+) cells in newly diagnosed ITP patients [(15.309 ± 10.756)%] and in ITP patients with recovery stage [(18.242 ± 12.243)%] were significantly higher than that in controls [(8.618 ± 5.719) %] (P < 0.05). The ratio of intracytoplasm CD4(+) CXCR5(+) IL-21(+)/CD4(+)CXCR5(+) cells in newly diagnosed ITP patients [(58.560 ± 26.285)%] and in ITP patients with recovery stage [(57.035 ± 30.936)%] were significantly higher than that in controls [(36.289 ± 24.868)%] (P < 0.05). The relative expression levels of BCL-6 mRNA in BMMNC of three groups were (1.407 ± 0.264), (1.149 ± 0.217) and (0.846 ± 0.157), respectively. The differences between 3 groups were significant(P < 0.05). It is concluded that the quantity and function of Tfh cells in ITP patients increase, which may play an important role in the pathogenesis of ITP.

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