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Sample records for lysed cells indicating

  1. 21 CFR 864.8540 - Red cell lysing reagent.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Red cell lysing reagent. 864.8540 Section 864.8540...) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Reagents § 864.8540 Red cell lysing reagent. (a) Identification. A red cell lysing reagent is a device used to lyse (destroy) red blood cells...

  2. Lyse theses; Lyse teser

    Energy Technology Data Exchange (ETDEWEB)

    Gjelsvik, Martin

    2012-07-01

    The energy sector is undergoing major changes: demand for energy is growing, especially outside Europe in the emerging economies. At the same time the energy production increases pollutants that harm the environment. Despite the construction of more renewable energy, fossil fuels play a dominant role in the years ahead. The owners and the board of Lyse Group has since its establishment in 1999 developed new business areas based on the Group's core competencies. Energy and telecommunications are currently the mainstays of the group, and a growing portion of its revenues from the telecom sector. The historical basis for Lyse has been hydropower production and distribution of electricity in the region - which still make up the bulk of the business. But industry drift has been great: Today Lyse is a major energy company with almost 300 000 paired fiber broadband customers throughout Norway. Lyse owners indicates a long-term industrial perspective, which has provided the opportunity for large investments: Lyse have in 2012 a power grid where 75% is in the ground, a gas network that can accommodate biogas, a district heating network that covers a large area of commercial space and a fiber network covering much of the Stavanger region. Lyse is working to develop new business models. The goal is to create smarter grids and smart home solutions through connection of business areas such as energy and telecommunications. The board of Lyse has asked IRIS develop a set of guiding principles that can provide a platform for strategy development in Lyse. The order from Lyse has not been a desire for the 'politically correct' or scenarios about the future. A thesis can on the contrary be considered as research-based assertion, based on what the research communities and experts believe is 'true' today. The report in its entirety will be available from the 8. of October 2012. (eb)

  3. Ultrasensitive detection of cell lysing in an microfabricated semiconductor laser cavity

    Energy Technology Data Exchange (ETDEWEB)

    Gourley, P.L.; French, T.; McDonald, A.E.; Shields, E.A. [Sandia National Labs., Albuquerque, NM (United States); Gourley, M.F. [Washington Hospital Center, Washington, DC (United States)

    1998-01-01

    In this paper the authors report investigations of semiconductor laser microcavities for use in detecting changes of human blood cells during lysing. By studying the spectra before and during mixing of blood fluids with de-ionized water, they are able to quantify the cell shape and concentration of hemoglobin in real time during the dynamical process of lysing. The authors find that the spectra can detect subtle changes that are orders of magnitude smaller than can be observed by standard optical microscopy. Such sensitivity in observing cell structural changes has implications for measuring cell fragility, monitoring apoptotic events in real time, development of photosensitizers for photodynamic therapy, and in-vitro cell micromanipulation techniques.

  4. Extraction and fractionation of RNA and DNA from single cells using selective lysing and isotachophoresis

    Science.gov (United States)

    Shintaku, Hirofumi; Santiago, Juan G.

    2015-03-01

    Single cell analyses of RNA and DNA are crucial to understanding the heterogeneity of cell populations. The numbers of approaches to single cells analyses are expanding, but sequence specific measurements of nucleic acids have been mostly limited to studies of either DNA or RNA, and not both. This remains a challenge as RNA and DNA have very similar physical and biochemical properties, and cross-contamination with each other can introduce false positive results. We present an electrokinetic technique which creates the opportunity to fractionate and deliver cytoplasmic RNA and genomic DNA to independent downstream analyses. Our technique uses an on-chip system that enables selective lysing of cytoplasmic membrane, extraction of RNA (away from genomic DNA and nucleus), focusing, absolute quantification of cytoplasmic RNA mass. The absolute RNA mass quantification is performed using fluorescence observation without enzymatic amplification in genomic DNA amount in the nucleus can be measured. We demonstrate the technique using single mouse B lymphocyte cells, for which we extracted an average of 14.1 pg total cytoplasmic RNA per cell. We also demonstrate correlation analysis between the absolute amount of cytoplasmic RNA and relative amount of genomic DNA, showing heterogeneity associated with cell cycle.

  5. A rapid chemical method for lysing Arabidopsis cells for protein analysis

    Directory of Open Access Journals (Sweden)

    Takano Tetsuo

    2011-07-01

    Full Text Available Abstract Background Protein extraction is a frequent procedure in biological research. For preparation of plant cell extracts, plant materials usually have to be ground and homogenized to physically break the robust cell wall, but this step is laborious and time-consuming when a large number of samples are handled at once. Results We developed a chemical method for lysing Arabidopsis cells without grinding. In this method, plants are boiled for just 10 minutes in a solution containing a Ca2+ chelator and detergent. Cell extracts prepared by this method were suitable for SDS-PAGE and immunoblot analysis. This method was also applicable to genomic DNA extraction for PCR analysis. Our method was applied to many other plant species, and worked well for some of them. Conclusions Our method is rapid and economical, and allows many samples to be prepared simultaneously for protein analysis. Our method is useful not only for Arabidopsis research but also research on certain other species.

  6. Experimental and theoretical study of hydrodynamic cell lysing of cancer cells in a high-throughput Circular Multi-Channel Microfiltration device

    KAUST Repository

    Ma, W.

    2013-04-01

    Microfiltration is an important microfluidic technique suitable for enrichment and isolation of cells. However, cell lysing could occur due to hydrodynamic damage that may be detrimental for medical diagnostics. Therefore, we conducted a systematic study of hydrodynamic cell lysing in a high-throughput Circular Multi-Channel Microfiltration (CMCM) device integrated with a polycarbonate membrane. HeLa cells (cervical cancer cells) were driven into the CMCM at different flow rates. The viability of the cells in the CMCM was examined by fluorescence microscopy using Acridine Orange (AO)/Ethidium Bromide (EB) as a marker for viable/dead cells. A simple analytical cell viability model was derived and a 3D numerical model was constructed to examine the correlation of between cell lysing and applied shear stress under varying flow rate and Reynolds number. The measured cell viability as a function of the shear stress was consistent with theoretical and numerical predictions when accounting for cell size distribution. © 2013 IEEE.

  7. The Bacillus thuringiensis vegetative insecticidal protein Vip3A lyses midgut epithelium cells of susceptible insects.

    Science.gov (United States)

    Yu, C G; Mullins, M A; Warren, G W; Koziel, M G; Estruch, J J

    1997-02-01

    The Vip3A protein is a member of a newly discovered class of vegetative insecticidal proteins with activity against a broad spectrum of lepidopteran insects. Histopathological observations indicate that Vip3A ingestion by susceptible insects such as the black cutworm (Agrotis ipsilon) and fall armyworm (Spodoptera frugiperda) causes gut paralysis at concentrations as low as 4 ng/cm2 of diet and complete lysis of gut epithelium cells resulting in larval death at concentrations above 40 ng/cm2. The European corn borer (Ostrinia nubilalis), a nonsusceptible insect, does not develop any pathology upon ingesting Vip3A. While proteolytic processing of the Vip3A protein by midgut fluids obtained from susceptible and nonsusceptible insects is comparable, in vivo immunolocalization studies show that Vip3a binding is restricted to gut cells of susceptible insects. Therefore, the insect host range for Vip3A seems to be determined by its ability to bind gut cells. These results indicate that midgut epithelium cells of susceptible insects are the primary target for the Vip3A insecticidal protein and that their subsequent lysis is the primary mechanism of lethality. Disruption of gut cells appears to be the strategy adopted by the most effective insecticidal proteins.

  8. Concerning the role of cell lysis-cryptic growth in anaerobic side-stream reactors: the single-cell analysis of viable, dead and lysed bacteria.

    Science.gov (United States)

    Foladori, P; Velho, V F; Costa, R H R; Bruni, L; Quaranta, A; Andreottola, G

    2015-05-01

    In the Anaerobic Side-Stream Reactor (ASSR), part of the return sludge undergoes alternating aerobic and anaerobic conditions with the aim of reducing sludge production. In this paper, viability, enzymatic activity, death and lysis of bacterial cells exposed to aerobic and anaerobic conditions for 16 d were investigated at single-cell level by flow cytometry, with the objective of contributing to the understanding of the mechanisms of sludge reduction in the ASSR systems. Results indicated that total and viable bacteria did not decrease during the anaerobic phase, indicating that anaerobiosis at ambient temperature does not produce a significant cell lysis. Bacteria decay and lysis occurred principally under aerobic conditions. The aerobic decay rate of total bacteria (bTB) was considered as the rate of generation of lysed bacteria. Values of bTB of 0.07-0.11 d(-1) were measured in anaerobic + aerobic sequence. The enzymatic activity was not particularly affected by the transition from anaerobiosis to aerobiosis. Large solubilisation of COD and NH4(+) was observed only under anaerobic conditions, as a consequence of hydrolysis of organic matter, but not due to cell lysis. The observations supported the proposal of two independent mechanisms contributing equally to sludge reduction: (1) under anaerobic conditions: sludge hydrolysis of non-bacterial material, (2) under aerobic conditions: bacterial cell lysis and oxidation of released biodegradable compounds.

  9. The Bacillus thuringiensis vegetative insecticidal protein Vip3A lyses midgut epithelium cells of susceptible insects.

    OpenAIRE

    Yu, C G; Mullins, M A; Warren, G W; Koziel, M G; Estruch, J J

    1997-01-01

    The Vip3A protein is a member of a newly discovered class of vegetative insecticidal proteins with activity against a broad spectrum of lepidopteran insects. Histopathological observations indicate that Vip3A ingestion by susceptible insects such as the black cutworm (Agrotis ipsilon) and fall armyworm (Spodoptera frugiperda) causes gut paralysis at concentrations as low as 4 ng/cm2 of diet and complete lysis of gut epithelium cells resulting in larval death at concentrations above 40 ng/cm2....

  10. Process for inhibiting the growth of a culture of lactic acid bacteria, and optionally lysing the bacterial cells, and uses of the resulting lysed culture

    NARCIS (Netherlands)

    Nauta, Arjen; Venema, Gerard; Kok, Jan; Ledeboer, Aat M.

    1995-01-01

    The invention provides a process for inhibiting the growth of a culture of lactic acid bacteria, or a product containing such culture e.g. a cheese product, in which in the cells of the lactic acid bacteria a holin obtainable from bacteriophages of Gram-positive bacteria, esp. from bacteriophages of

  11. Cascade cell lyses and DNA extraction for identification of genes and microorganisms in kefir grains.

    Science.gov (United States)

    Kowalczyk, Magdalena; Kolakowski, Piotr; Radziwill-Bienkowska, Joanna M; Szmytkowska, Agnieszka; Bardowski, Jacek

    2012-02-01

    Kefir is a dairy product popular in many countries in Central Europe, especially in Poland and other countries of Eastern and Northern Europe. This type of fermented milk is produced by a complex population of symbiotic bacteria and yeasts. In this work, conditions for DNA extraction, involving disruption of kefir grains and a cascade of cell lysis treatments, were established. Extraction procedure of total microbial DNA was carried out directly from fresh kefir grains. Using different lysis stringency conditions, five DNA pools were obtained. Genetic diversity of DNA pools were validated by RAPD analysis, which showed differences in patterns of amplified DNA fragments, indicating diverse microbial composition of all the analysed samples. These DNA pools were used for construction of genomic DNA libraries for sequencing. As much as 50% of the analysed nucleotide sequences showed homology to sequences from bacteria belonging to the Lactobacillus genus. Several sequences were similar to sequences from bacteria representing Lactococcus, Oenococcus, Pediococcus, Streptococcus and Leuconostoc species. Among homologues of yeast proteins were those from Candida albicans, Candida glabrata, Kluyveromyces lactis and Saccharomyces cerevisiae. In addition, several sequences were found to be homologous to sequences from bacteriophages.

  12. Tumor Lysing Genetically Engineered T Cells Loaded with Multi-Modal Imaging Agents

    Science.gov (United States)

    Bhatnagar, Parijat; Alauddin, Mian; Bankson, James A.; Kirui, Dickson; Seifi, Payam; Huls, Helen; Lee, Dean A.; Babakhani, Aydin; Ferrari, Mauro; Li, King C.; Cooper, Laurence J. N.

    2014-03-01

    Genetically-modified T cells expressing chimeric antigen receptors (CAR) exert anti-tumor effect by identifying tumor-associated antigen (TAA), independent of major histocompatibility complex. For maximal efficacy and safety of adoptively transferred cells, imaging their biodistribution is critical. This will determine if cells home to the tumor and assist in moderating cell dose. Here, T cells are modified to express CAR. An efficient, non-toxic process with potential for cGMP compliance is developed for loading high cell number with multi-modal (PET-MRI) contrast agents (Super Paramagnetic Iron Oxide Nanoparticles - Copper-64; SPION-64Cu). This can now be potentially used for 64Cu-based whole-body PET to detect T cell accumulation region with high-sensitivity, followed by SPION-based MRI of these regions for high-resolution anatomically correlated images of T cells. CD19-specific-CAR+SPIONpos T cells effectively target in vitro CD19+ lymphoma.

  13. A novel assay of biofilm antifungal activity reveals that amphotericin B and caspofungin lyse Candida albicans cells in biofilms.

    Science.gov (United States)

    DiDone, Louis; Oga, Duana; Krysan, Damian J

    2011-08-01

    The ability of Candida albicans to form drug-resistant biofilms is an important factor in its contribution to human disease. Assays to identify and characterize molecules with activity against fungal biofilms are crucial for the development of drugs with improved anti-biofilm activity. Here we report the application of an adenylate kinase (AK)-based cytotoxicity assay of fungal cell lysis to the characterization of agents active against C. albicans biofilms. We have developed three protocols for the AK assay. The first measures AK activity in the supernatants of biofilms treated with antifungal drugs and can be performed in parallel with a standard 2,3-bis-(2-methoxy-4-nitro-5-sulphophenyl)-2H-tetrazolium-5-caboxanilide-based biofilm susceptibility assay; a second, more sensitive protocol measures the AK activity present within the biofilm matrix; and a third procedure allows the direct visualization of lytic activity toward biofilms formed on catheter material. Amphotericin B and caspofungin, the two most effective anti-biofilm drugs currently used to treat fungal infections, both directly lyse planktonic C. albicans cells in vitro, leading to the release of AK into the culture medium. These studies serve to validate the AK-based lysis assay as a useful addition to the methods for the characterization of antifungal agents active toward biofilms and provide insights into the mode of action of amphotericin B and caspofungin against C. albicans biofilms. Copyright © 2011 John Wiley & Sons, Ltd.

  14. Evidence for the formation of endothelin by lysed red blood cells from endogenous precursor.

    Science.gov (United States)

    Tippler, B; Herbst, C; Simmet, T

    1994-12-12

    The release of endothelin from various blood cell fractions was investigated. Human as well as rat blood cell fractions homogenized by sonification were incubated in buffer for up to 60 min. Neither in platelet nor leukocyte homogenates from either species could immunoreactive endothelin be detected. In contrast, homogenates of red blood cells from both species showed a rapid and time-dependent rise of immunoreactive endothelin levels, reaching a peak at 15 min and decreasing thereafter. However, at time point 0 no immunoreactive endothelin could be detected. Reverse phase high performance liquid chromatography showed immunoreactive endothelin to consist of endothelin-1 as well as big endothelin-1. The release of immunoreactive endothelin in human and rat homogenates was concentration-dependently inhibited by the protease inhibitors, leupeptin, phosphoramidon, chymostatin and pepstatin A in order of increasing potency. Intact red blood cells did not incorporate [125I]endothelin-1 nor did they transform exogenous big endothelin-1 to endothelin-1. However, haemolysis of red blood cells with hypotonic saline (0.2%) or incubation with pore-forming staphylococcal alpha-toxin induced the release of immunoreactive endothelin into the buffer samples. Thus, apart from the indirect vasoconstrictor, haemoglobin, red blood cells can also liberate the direct vasoconstrictor, endothelin, a finding expected to be of considerable pathophysiological significance.

  15. PENGARUH EKSTRAK JAMU TERHADAP AKTIVITAS SEL NATURAL KILLER DALAM MELISIS ALUR SEL LEUKIMIA (K-562 SECARA IN VITRO [The Effects of Commercial “Jamu” Extracts on Natural Killer Cell Activity in Lysing Leukemic Cell Line (K-562 in vitro

    Directory of Open Access Journals (Sweden)

    Elisa Veronica D.C. 2

    2002-04-01

    Full Text Available Natural killer (NK cell consitutes white blood cells which specifically functions in lysing tumor and virus invected cells. In this research, a commercial “Jamu” was tested to observe its effect on NK cells activity against leukemic cell lines (K562 in vitro. Jamu was extracted with hot water, diluted and added into cell cultures consisted of a mixture of human peripheric limphocyte cells, as the source of the effector NK cells, and K562 cell line i.e., the target cells which were cell line derived from human leukemia and had been labelled with H3-thymidine. The mixture of the cells were made by culturing the two cells at the ratio of 50:1 and 100 : 1, respectively. The results showed that lysing activity of NK cells in the presence of “Jamu” water extract measured as lysing percentage and lysing index increased only slightly, which were not statiscally significant. It should be considered that the test used in this research represents only a part of the lysing mechanism by NK cells against the target cells. An in vivo test for a period of time will be recessary to elucidate ffurther this NK cell activity.

  16. Bacteriophage enzymes for the prevention and treatment of bacterial infections: Stability and stabilization of the enzyme lysing Streptococcus pyogenes cells

    Energy Technology Data Exchange (ETDEWEB)

    Klyachko, N. L.; Dmitrieva, N. F.; Eshchina, A. S.; Ignatenko, O. V.; Filatova, L. Y.; Rainina, Evguenia I.; Kazarov, A. K.; Levashov, A. V.

    2008-06-01

    Recombinant, phage associated lytic enzyme Ply C capable to lyse streptococci of groups A and C was stabilized in the variety of the micelles containing compositions to improve the stability of the enzyme for further application in medicine. It was shown that, in the micellar polyelectrolyte composition M16, the enzyme retained its activity for 2 months; while in a buffer solution under the same conditions ((pH 6.3, room temperature), it completely lost its activity in 2 days

  17. Cancer Patient T Cells Genetically Targeted to Prostate-Specific Membrane Antigen Specifically Lyse Prostate Cancer Cells and Release Cytokines in Response to Prostate-Specific Membrane Antigen

    Directory of Open Access Journals (Sweden)

    Michael C. Gong

    1999-06-01

    Full Text Available The expression of immunoglobulin-based artificial receptors in normal T lymphocytes provides a means to target lymphocytes to cell surface antigens independently of major histocompatibility complex restriction. Such artificial receptors have been previously shown to confer antigen-specific tumoricidal properties in murine T cells. We constructed a novel ζ chain fusion receptor specific for prostate-specific membrane antigen (PSMA termed Pz-1. PSMA is a cell-surface glycoprotein expressed on prostate cancer cells and the neovascular endothelium of multiple carcinomas. We show that primary T cells harvested from five of five patients with different stages of prostate cancer and transduced with the Pz-1 receptor readily lyse prostate cancer cells. Having established a culture system using fibroblasts that express PSMA, we next show that T cells expressing the Pz-1 receptor release cytokines in response to cell-bound PSMA. Furthermore, we show that the cytokine release is greatly augmented by B7.1-mediated costimulation. Thus, our findings support the feasibility of adoptive cell therapy by using genetically engineered T cells in prostate cancer patients and suggest that both CD4+ and CD8+ T lymphocyte functions can be synergistically targeted against tumor cells.

  18. TCR gamma delta cytotoxic T lymphocytes expressing the killer cell-inhibitory receptor p58.2 (CD158b) selectively lyse acute myeloid leukemia cells.

    Science.gov (United States)

    Dolstra, H; Fredrix, H; van der Meer, A; de Witte, T; Figdor, C; van de Wiel-van Kemenade, E

    2001-05-01

    Cytotoxic T lymphocytes (CTL) are thought to play an important role in the graft-versus-leukemia (GVL) response. Unfortunately, GVL reactivity is often associated with life-threatening graft-versus-host disease (GVHD). Characterization of CTL that selectively attack leukemic cells but not normal cells may lead to the development of adjuvant immunotherapy that separates GVL from GVHD. Here, we describe TCR gamma delta (V gamma 9/V delta 1) CTL, isolated from the peripheral blood of an AML patient after stem cell transplantation (SCT), that very efficiently lysed freshly isolated acute myeloid leukemia (AML) cells and AML cell lines. Interestingly, HLA-matched non-malignant hematopoietic cells were not killed. We revealed that the killer cell-inhibitory receptor (KIR) p58.2 (CD158b) specific for group 2 HLA-C molecules negatively regulates the cytotoxic effector function displayed by these TCR gamma delta CTL. First, an antibody against HLA-C enhances lysis of non-malignant cells. Secondly, stable transfection of HLA-Cw*0304 into the class I-negative cell line 721.221 inhibited lysis. Finally, engagement of p58.2 by antibodies immobilized on Fc gamma R-expressing murine P815 cells inhibits CD3- and TCR gamma delta-directed lysis. Compared to non-malignant hematopoietic cells, AML cells express much lower levels of MHC class I molecules making them susceptible to lysis by p58.2(+) TCR gamma delta CTL. Such KIR-regulated CTL reactivity may have a role in the GVL response without affecting normal tissues of the host and leading to GVHD.

  19. Further characterization of particulate fractions from lysed cell envelopes of Halobacterium halobium and isolation of gas vacuole membranes.

    Science.gov (United States)

    Toeckenius, W; Kunau, W H

    1968-08-01

    Lysates of cell envelopes from Halobacterium halobium have been separated into four fractions. A soluble, colorless fraction (I) containing protein, hexosamines, and no lipid is apparently derived from the cell wall. A red fraction (II), containing approximately 40 per cent lipid, 60 per cent protein, and a small amount of hexosamines consists of cell membrane disaggregated into fragments of small size. A third fraction (III) of purple color consists of large membrane sheets and has a very similar composition to II, containing the same classes of lipids but no hexosamines; its buoyant density is 1.18 g/ml. The fourth fraction (IV) has a buoyant density of 1.23 g/ml and contains the "intracytoplasmic membranes." These consist mainly of protein, and no lipid can be extracted with chloroform-methanol. Fractions I and II, which result from disaggregation of cell wall and cell membrane during lysis, contain a high proportion of dicarboxyl amino acids; this is in good agreement with the assumption that disruption of the cell envelope upon removal of salt is due to the high charge density. The intracytoplasmic membranes (IV) represent the gas vacuole membranes in the collapsed state. In a number of mutants that have lost the ability to form gas vacuoles, no vacuole membranes or any structure that could be related to them has been found.

  20. Listeria monocytogenes that lyse in the macrophage cytosol trigger AIM2-mediated pyroptosis

    Science.gov (United States)

    Sauer, John-Demian; Witte, Chelsea E.; Zemansky, Jason; Hanson, Bill; Lauer, Peter; Portnoy, Daniel A.

    2010-01-01

    Summary To gain insight into the mechanisms by which host cells detect cytosolic invasion by intracellular pathogens, a genetic screen was performed to identify Listeria monocytogenes mutants that induced altered levels of host cell death. A mutation in lmo2473 resulted in hyper-stimulation of host cell death and IL-1β secretion (pyroptosis) following bacteriolysis in the macrophage cytosol. In addition, strains engineered to lyse in the cytosol by expression of both bacteriophage holin and lysin or induced to lyse by treatment with ampicillin stimulated pyroptosis. Pyroptosis was independent of the Nlrp3 and Nlrc4 receptors, but dependent on ASC and AIM2. Importantly, wild type L. monocytogenes were also found to lyse, albeit at low levels, and trigger AIM2-dependent pyroptosis. Since AIM2 is activated by DNA, these data suggested that pyroptosis is triggered by bacterial DNA released during lysis. PMID:20417169

  1. Cytotoxic T-lymphocyte clones, established by stimulation with the HLA-A2 binding p5365-73 wild type peptide loaded on dendritic cells In vitro, specifically recognize and lyse HLA-A2 tumour cells overexpressing the p53 protein

    DEFF Research Database (Denmark)

    Barfoed, Annette Malene; Petersen, T R; Kirkin, A F;

    2000-01-01

    to carry identical T-cell receptors. The CTL clone, 2D9, was shown to specifically lyse the HLA-A*0201+ squamous carcinoma cell line SCC9 and the breast cancer cell line MDA-MB-468. Our data demonstrate that human peripheral blood lymphocytes from normal healthy individuals comprise T cells capable...... of recognizing p53 derived wild type (self) peptides. Furthermore, the capacity of R9V specific T cell clones to exert HLA restricted cytotoxicity, argues that the R9V peptide is naturally presented on certain cancer cells. This supports the view that p53 derived wild type peptides might serve as candidate...... target antigens for the immunotherapeutic treatment of cancer....

  2. Lytic Characteristics and Identification of Two Alga-lysing Bacterial Strains

    Institute of Scientific and Technical Information of China (English)

    PEI Haiyan; HU Wenrong

    2006-01-01

    All previously reported bacterial species which are capable of lysing harmful algae have been isolated from coastal environments in which harmful algae blooms have occurred. Due to the low concentration of alga-lysing bacteria in an algal bloom, it is difficult to isolate the alga-lysing bacteria by existing methods. In this paper, two algae-lysing bacterial strains,P01 and P03, have been isolated from a biosystem immobilized on a sponge that was highly effective in removing algae and microcystins. Their lysing modes and effects on Microcystis aeruginosa have been studied. The results show that the degradation processes of these two strains for M. aeruginosa accorded with a first-order reaction model when the chlorophylla concentration was in the range from 0 to 1000 μg L-1. The degradation rate constants were 0.106 7, 0.127 4 and 0.279 2 for P01and0.0683, 0.0744 and 0.02897 for P03, when the bacterial densities were 8.6 × 105, 8.6 × 106 and 8.6 × 107cells mL 1, respectively. Moreover, the two bacterial strains had favourable lytic effects not only on M. aeruginosa, but also on Chlorella and Scene-desmus. Their lytic effect on M. aeruginosa did not require physical cell to cell contact, but proceeded by the production of an extracellular product. The bacterial strains were identified as Bacillus species by PCR amplification of the 16S rRNA gene, BLAST analysis, and comparison with sequences in the GenBank nucleotide database.

  3. Listeria monocytogenes that lyse in the macrophage cytosol trigger AIM2-mediated pyroptosis

    OpenAIRE

    Sauer, John-Demian; Witte, Chelsea E.; Zemansky, Jason; Hanson, Bill; Lauer, Peter; Portnoy, Daniel A.

    2010-01-01

    To gain insight into the mechanisms by which host cells detect cytosolic invasion by intracellular pathogens, a genetic screen was performed to identify Listeria monocytogenes mutants that induced altered levels of host cell death. A mutation in lmo2473 resulted in hyper-stimulation of host cell death and IL-1β secretion (pyroptosis) following bacteriolysis in the macrophage cytosol. In addition, strains engineered to lyse in the cytosol by expression of both bacteriophage holin and lysin or ...

  4. Listeria monocytogenes that lyse in the macrophage cytosol trigger AIM2-mediated pyroptosis

    OpenAIRE

    Sauer, John-Demian; Witte, Chelsea E.; Zemansky, Jason; Hanson, Bill; Lauer, Peter; Portnoy, Daniel A.

    2010-01-01

    To gain insight into the mechanisms by which host cells detect cytosolic invasion by intracellular pathogens, a genetic screen was performed to identify Listeria monocytogenes mutants that induced altered levels of host cell death. A mutation in lmo2473 resulted in hyper-stimulation of host cell death and IL-1β secretion (pyroptosis) following bacteriolysis in the macrophage cytosol. In addition, strains engineered to lyse in the cytosol by expression of both bacteriophage holin and lysin or ...

  5. Isolation and algae-lysing characteristics of the algicidal bacterium B5

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Water blooms have become a worldwide environmental problem. Recently, algicidal bacteria have attracted wide attention as possible agents for inhibiting algal water blooms. In this study, one strain of algicidal bacterium B5 was isolated from activated sludge. On the basis of analysis of its physiological characteristics and 16S rDNA gene sequence, it was identified as Bacillus fusiformis. Its algae-lysing characteristics on Microcystis aeruginosa, Chlorella and Scenedesmus were tested. The results showed that: (1) the algicidal bacterium B5 is a Gram-negative bacterium. The 16S rDNA nucleotide sequence homology of strain B5 with 2 strains of B. fusiformis reached 99.86%, so B5 was identified as B. fusiformis; (2) the algal-lysing effects of the algicidal bacterium B5 on M. aeruginosa, Chlorella and Scenedesmus were pronounced. The initial bacterial and algal cell densities strongly influence the removal rates of chlorophyll-a. The greater the initial bacterial cell density, the faster the degradation of chlorophyll-a. The greater the initial algal cell density, the slower the degradation of chlorophyll-a. When the bacterial cell density was 3.6 × 107 cells/ml, nearly 90% of chlorophyll-a was removed. When the chlorophyll-a concentration was less than 550 μg/L, about 70 % was removed; (3) the strain B5 lysed algae not directly but by secreting metabolites and these metabolites could bear heat treatment.

  6. Alga-lysing bioreactor and the dominant bacteria strain

    Institute of Scientific and Technical Information of China (English)

    PEI Hai-yan; HU Wen-rong; MU Rui-min; LI Xiao-cai

    2007-01-01

    Alga-lysing bacteria have been paid much attention to in recent years. In this study, the alga-lysing strain P05 which was isolated from an immobilizing biosystem was immobilized by coke and elastic filler, forming two biological reactors. The removal efficiencies of algae, NH3-N and organic matter using the two reactors were studied. The results showed that strain P05 was an ideal algal-lysing bacteria strain because it was easy to be immobilized by coke and elastic filler which are of cheap, low biodegradability and the simple immobilization procedure. After 7 d filming, the biological film could be formed and the reactors were used to treat the eutrophic water. These two reactors were of stability and high effect with low cost and easy operation. The optimal hydraulic retention time (HRT) of each reactor was 4 h. The algae removal rates were 80.38% and 82.1% (in term of Chl-a) of coke reactor and filler reactor, respectively. And that of NH3-N were 52.3% and 52.7%. The removal rates of CODMn were 39.03% and 39.64%. The strain P05 was identified as Bacillus sp. by PCR amplification of the 16S rRNA gene, BLAST analysis, and comparison with sequences in the GenBank nucleotide database.

  7. Lake viruses lyse cyanobacteria, Cylindrospermopsis raciborskii, enhances filamentous-host dispersal in Australia

    Science.gov (United States)

    Pollard, Peter C.; Young, Loretta M.

    2010-01-01

    Globally, cyanobacterial blooms are increasing along with observations of the controlling influence of viruses. Our aim here was to test whether viruses from an Australian freshwater lake could lyse the cyanobacterium Cylindrospermopsis raciborskii (Woloszynska) Seenaya and Subba Raju. C. raciborskii was selectively isolated from Lake Samsonvale southeast Queensland Australia using a Modified Jaworski Medium (without any form of inorganic nitrogen). Microscopy confirmed the resulting culture of a single cyanobacterial species. Natural viral-like particles (VLPs) were incubated with C. raciborskii cells, the host abundance decreased by 86% in 5 days, while the number of VLPs increased stepwise. As a cell lysed, the filaments of cells split into smaller, but viable, fragments. This process may help disperse the cyanobacterium in the wild. Hence the use of this virus to control blooms may inadvertently encourage the dispersal of toxic filamentous cyanobacteria. The cyanophage (virus infecting cyanobacteria) replication time was 21 h, with an average burst size of 64 viruses cell -1. Transmission Electron Microscopy showed this cyanophage for C. raciborskii, with its long, non-contractile tail and a capsid diameter of 70 nm, belongs to the Siphoviridae family of viruses. This cyanophage can affect the abundance and distribution of the cyanobacterium C. raciborskii in this Australian freshwater lake.

  8. Lysing bloom-causing alga Phaeocystis globosa with microbial algicide: An efficient process that decreases the toxicity of algal exudates.

    Science.gov (United States)

    Cai, Guanjing; Yang, Xujun; Lai, Qiliang; Yu, Xiaoqi; Zhang, Huajun; Li, Yi; Chen, Zhangran; Lei, Xueqian; Zheng, Wei; Xu, Hong; Zheng, Tianling

    2016-02-05

    Algicidal microbes could effectively remove the harmful algae from the waters. In this study, we were concerned with the ecological influence of an algicide extracted from Streptomyces alboflavus RPS, which could completely lyse the Phaeocystis globosa cells within two days. In microcosms, 4 μg/mL of the microbial algicide could efficiently remove P. globosa cells without suppressing other aquatic organisms. Bioluminescent assays confirmed that the toxicity of microbial algicide at this concentration was negligible. Interestingly, the toxicity of P. globosa exudates was also significantly reduced after being treated with the algicide. Further experiments revealed that the microbial algicide could instantly increase the permeability of the plasma membrane and disturb the photosynthetic system, followed by the deformation of organelles, vacuolization and increasing oxidative stress. The pre-incubation of N-acetyl cysteine (NAC) verified that the rapid damages to the plasma membrane and photosynthetic system caused the algal death in the early phase, and the increasing oxidative stress killed the rest. The late accumulation and possible release of CAT also explained the decreasing toxicity of the algal culture. These results indicated that this microbial algicide has great potential in controlling the growth of P. globosa on site.

  9. Attenuating l-lysine production by deletion of ddh and lysE and their effect on l-threonine and l-isoleucine production in Corynebacterium glutamicum.

    Science.gov (United States)

    Dong, Xunyan; Zhao, Yue; Hu, Jinyu; Li, Ye; Wang, Xiaoyuan

    2016-11-01

    The fermentative production of l-threonine and l-isoleucine with Corynebacterium glutamicum is usually accompanied by the by-production of l-lysine, which shares partial biosynthesis pathway with l-threonine and l-isoleucine. Since the direct precursor for l-lysine synthesis, diaminopimelate, is a component of peptidoglycan and thus essential for cell wall synthesis, reducing l-lysine by-production could be troublesome. Here, a basal strain with eliminated l-lysine production was constructed from the wild type C. glutamicum ATCC13869 by deleting the chromosomal ddh and lysE. Furthermore, the basal strain as well as the ddh single mutant strain was engineered for l-threonine production by over-expressing lysC1, hom1 and thrB, and for l-isoleucine production by over-expressing lysC1, hom1, thrB and ilvA1. Fermentation experiments with the engineered strains showed that (i) deletion of ddh improved l-threonine production by 17%, and additional deletion of lysE further improved l-threonine production by 28%; (ii) deletion of ddh improved l-isoleucine production by 8% and improved cell growth by 21%, whereas additional deletion of lysE had no further influence on both l-isoleucine production and cell growth; (iii) l-lysine by-production was reduced by 95% and 86% in l-threonine and l-isoleucine production, respectively, by deletion of ddh and lysE. This is the first report on improving l-threonine and l-isoleucine production by deleting ddh and lysE in C. glutamicum. The results demonstrate deletion of ddh and lysE as an effective strategy to reduce l-lysine by-production without surrendering the cell growth of C. glutamicum.

  10. 21 CFR 864.5300 - Red cell indices device.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Red cell indices device. 864.5300 Section 864.5300....5300 Red cell indices device. (a) Identification. A red cell indices device, usually part of a larger... corpuscular hemoglobin (MCH), and the mean corpuscular hemoglobin concentration (MCHC). The red cell...

  11. Miniaturized Cell Lysis Device Using Spherically Focused Ultrasound

    Institute of Scientific and Technical Information of China (English)

    李刚; 肖宏; 郭旻; 程京

    2003-01-01

    A prototype of a miniaturized cell lysis device developed using a concave spherical transducer is capable of lysing bacteria without added chemical denaturants, enzymes or microparticles and is capable of efficiently lysing yeast without any mechanical or enzymatic pretreatment.The device is designed for miniature bio-analysis systems where cell lysing is needed to obtain intracellular materials for further analysis such as DNA identification.The device lysis efficiency was evaluated using viable cell counts and microscopy.Additionally, the device efficiency was compared with that of traditional chemical cell lysis methods using standard molecular biological techniques such as agarose gels and ultraviolet (UV) spectroscopy.The results indicate that efficient bacteria and cell disruption can be achieved through a low-voltage-driven and spherically focused high-frequency ultrasonic device.

  12. Measuring cell viability with membrane impermeable zinc fluorescent indicator.

    Science.gov (United States)

    Stork, Christian J; Li, Yang V

    2006-09-15

    Recent findings suggest that the accumulation of cytoplasmic zinc [Zn2+]i is a ubiquitous component in the cell death cascade. Zn2+ can be liberated from intracellular stores following oxidative stress and contribute to cell death processes. Here we show that the membrane/cell impermeable Zn2+ fluorescent indicator Newport Green (NG), which is non-toxic and impermeable to the membranes of healthy cells, can label unhealthy cells in tissue slices in a manner comparable to the traditional viability indicator propidium iodide (PI). Using confocal microscopy, we detected PI labeled nuclei colocalized with NG fluorescence. Our results indicate that cells which absorbed PI into their nuclei also allowed cell-impermeable Zn2+ dye to penetrate their plasma membranes, subsequently exhibiting cytosolic and nuclear fluorescence. As in PI staining, we observed marked increases in NG fluorescence in damaged/dead cells of tissue slices. Two other cell impermeable fluorescent Zn2+ dyes, Fluozin-3 and Zinpyr-4, also stained cytosolic Zn2+ in PI labeled cells. Our data indicates that the application of a Zn2+ fluorescent indicator is a fast, simple, non-toxic and reliable method for visualizing cell viability within in vitro tissue preparations. Accordingly, we demonstrate that intracellular accumulation of Zn2+ correlates with neuronal death.

  13. Physicochemical characterization of Staphylococcus aureus-lysing LysK enzyme in complexes with polycationic

    Science.gov (United States)

    Staphylococcus aureus causes many serious visceral, skin, and respiratory diseases. About 90% of clinical strains are multi-drug resistant, but the use of bacteriophage lytic enzymes offers a viable alternative to antibiotic therapy. LysK, the phage K endolysin can lyse S. aureus when purified and ...

  14. Staphylococcus aureus Targets the Duffy Antigen Receptor for Chemokines (DARC) to Lyse Erythrocytes

    NARCIS (Netherlands)

    Spaan, András N.; Reyes-Robles, Tamara; Badiou, Cédric; Cochet, Sylvie; Boguslawski, Kristina M.; Yoong, Pauline; Day, Christopher J.; Gosselaar-de Haas, Carla J C; van Kessel, Kok P M; Vandenesch, François; Jennings, Michael P.; Le Van Kim, Caroline; Colin, Yves; Van Strijp, Jos A G; Henry, Thomas; Torres, Victor J.

    2015-01-01

    In order for Staphylococcus aureus to thrive inside the mammalian host, the bacterium has to overcome iron scarcity. S. aureus is thought to produce toxins that lyse erythrocytes, releasing hemoglobin, the most abundant iron source in mammals. Here we identify the Duffy antigen receptor for chemokin

  15. THE EFFECTS OF OXIMES IN THE ASSAY OF ACETYLCHOLINESTERASE ACTIVITY IN LYSED ERYTHROCYTES IN VITRO

    Directory of Open Access Journals (Sweden)

    M. Abdollahi.

    1997-06-01

    Full Text Available Organophosphorus compounds are known to inhibit the esteratic site of acetylcholinesterase by phosphorylation. The phosphorylated esteratic site of acetylcholinesterase undergoes hydrolytic regeneration at a slow or negligible rate. Nucleophilic agents such as hydroxytamine, hydroxamic acids, and oximes reactivate the enzyme more erapidfy than does spontaneous hydrolysis. The red cell cholinesterose activity was assayed using dithio bis-2-nitrobenzoic acid (DTNB commonly known as Ellman's reagent. The principle of this assay method is the rate of hydrolysis of acetylthiocholine (substrate by a red celt suspension. Thiocholine that is produced, forms a yellow complex, when EUman's reagent (DTNB is used in the assay. This was tested in vitro in lysed erythrocyte samples of 35 healthy persons who had no known exposure to cholinesterose inhibitors, after the observation of immediate increase in absorption of light at 440 nm. All of data were statistically analyzed using one-way ANOVA and student t-test. A value of p<0.01 was considered. Results of this study show an increased absorbance in 440 nm, for pretreated samples with pratidoxime. This was observed by doses of (0.1, 0.5, 1,2 mmol, p<0.01. It was also a good dose dependent increase in absorbance at 440 nm for pralidoxime, (r=0.940, p<0.01. Also there is a significant increase in absorbance at 440 nm for samples pretreated by obidoxime at doses of (0.1, 0.5, 1,2 mmol. There is also a good correlation between absorbance at 440 nm and variou doses of obidoxime (r=0.946 , p<0.01. It is concluded that oximes can hydrofyzes the substrate, which then would be a source of error in determination of acetylcholinesterase activity and must be token into account.

  16. Combination of driselase and lysing enzyme in one molar potassium chloride is effective for the production of protoplasts from germinated conidia of Fusarium verticillioides.

    Science.gov (United States)

    Ramamoorthy, Vellaisamy; Govindaraj, Lavanya; Dhanasekaran, Madhumitha; Vetrivel, Sharmilee; Kumar, Krish K; Ebenezar, Edward

    2015-04-01

    Various cell wall degrading enzymes and the protoplasting media were evaluated for the production of protoplast in Fusarium verticillioides. Among the various enzymes tested, driselase at 12.5 mg/ml in 1 M KCl protoplasting medium produced the maximum number of protoplast. Next to driselase, lysing enzyme at 10 mg/ml in 1.2 M MgSO4 protoplasting medium was found to be the second best enzyme for the production of protoplast. More interestingly, the combined use of driselase @ 12.5 mg/ml and lysing enzyme @ 10 mg/ml in 1 M KCl exhibited the additive effect on protoplast formation. Germinated conidia of F. verticillioides are the most susceptible fungal material for protoplast production. The use of sucrose at 1.2 M in the regeneration medium supported the maximum regeneration of protoplast. From the present study, we recommend driselase (12.5 mg/ml) and lysing enzyme (10 mg/ml) in 1 M KCl protoplasting medium and germinated conidia of F. verticillioides for the maximum production of protoplasts and 1.2 M sucrose is the best osmoticum for the regeneration of protoplasts. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Automated Parameters for Troubled-Cell Indicators Using Outlier Detection

    NARCIS (Netherlands)

    Vuik, M.J.; Ryan, J.K.

    2016-01-01

    In Vuik and Ryan [J. Comput. Phys., 270 (2014), pp. 138--160] we studied the use of troubled-cell indicators for discontinuity detection in nonlinear hyperbolic partial differential equations and introduced a new multiwavelet technique to detect troubled cells. We found that these methods perform we

  18. Screening fluorescent voltage indicators with spontaneously spiking HEK cells.

    Directory of Open Access Journals (Sweden)

    Jeehae Park

    Full Text Available Development of improved fluorescent voltage indicators is a key challenge in neuroscience, but progress has been hampered by the low throughput of patch-clamp characterization. We introduce a line of non-fluorescent HEK cells that stably express NaV 1.3 and KIR 2.1 and generate spontaneous electrical action potentials. These cells enable rapid, electrode-free screening of speed and sensitivity of voltage sensitive dyes or fluorescent proteins on a standard fluorescence microscope. We screened a small library of mutants of archaerhodopsin 3 (Arch in spiking HEK cells and identified two mutants with greater voltage-sensitivity than found in previously published Arch voltage indicators.

  19. Gamma-H2AX biodosimetry for use in large scale radiation incidents: comparison of a rapid ‘96 well lyse/fix’ protocol with a routine method

    Directory of Open Access Journals (Sweden)

    Jayne Moquet

    2014-03-01

    Full Text Available Following a radiation incident, preliminary dose estimates made by γ-H2AX foci analysis can supplement the early triage of casualties based on clinical symptoms. Sample processing time is important when many individuals need to be rapidly assessed. A protocol was therefore developed for high sample throughput that requires less than 0.1 ml blood, thus potentially enabling finger prick sampling. The technique combines red blood cell lysis and leukocyte fixation in one step on a 96 well plate, in contrast to the routine protocol, where lymphocytes in larger blood volumes are typically separated by Ficoll density gradient centrifugation with subsequent washing and fixation steps. The rapid ‘96 well lyse/fix’ method reduced the estimated sample processing time for 96 samples to about 4 h compared to 15 h using the routine protocol. However, scoring 20 cells in 96 samples prepared by the rapid protocol took longer than for the routine method (3.1 versus 1.5 h at zero dose; 7.0 versus 6.1 h for irradiated samples. Similar foci yields were scored for both protocols and consistent dose estimates were obtained for samples exposed to 0, 0.2, 0.6, 1.1, 1.2, 2.1 and 4.3 Gy of 250 kVp X-rays at 0.5 Gy/min and incubated for 2 h. Linear regression coefficients were 0.87 ± 0.06 (R2 = 97.6% and 0.85 ± 0.05 (R2 = 98.3% for estimated versus actual doses for the routine and lyse/fix method, respectively. The lyse/fix protocol can therefore facilitate high throughput processing for γ-H2AX biodosimetry for use in large scale radiation incidents, at the cost of somewhat longer foci scoring times.

  20. An unconventional approach to impedance microbiology: detection of culture media conductivity variations due to bacteriophage generated lyses of host bacteria.

    Science.gov (United States)

    Mortari, Alessia; Adami, Andrea; Lorenzelli, Leandro

    2015-05-15

    A novel and unconventional approach to impedance microbiology has been under investigation. In our approach, solution conductivity variations are generated from bacteriophage lyses of infected host cells and the consequent release of conductive endoplasmic material. To sensitively detect the lysis, low conductive growth media have been developed. A microchip has been fabricated to perform the analysis. The microchip is made of two bare gold electrodes and PDMS microchamber of 36 nL volume. Escherichia coli and selective phages T4 have been used as case study. Proof-of-principle experiments are here presented and discussed. The method was characterised in a wide range between 10(4) and 10(8) CFU/mL, where linear relation was found between conductivity variation and cell concentration in a log10 vs. log10 plot. The method is suited to integration with sample preparation based on phage-functionalised magnetic beads. It has a potential detection limit below 1 CFU/chamber and a total assay time of less than 1 h.

  1. Morphological and molecular characterization of several actinophages isolated from soil which lyse Streptomyces cattleya or S. venezuelae.

    Science.gov (United States)

    Anné, J; Wohlleben, W; Burkardt, H J; Springer, R; Pühler, A

    1984-10-01

    Several lytic and lysogenic actinophages were isolated from soil samples infected with Streptomyces cattleya and S. venezuelae. The morphologies and some biological properties of the phages, and the physico-chemical characteristics of their DNAs, were compared. Electron micrographs indicated that all the phage heads were of an icosahedral form, but head size and length of the tail varied. Two of the phages had a broad host range; the other isolates could lyse only a limited number of species. The molecular sizes of the phage DNAs were between 32.2 and 98.5 kb as estimated by electron microscopy and restriction enzyme analysis. The same study also indicated that one of the DNA species contained cohesive ends. The G + C content of the DNAs ranged between 45.1 and 74.2 mol % as estimated from melting studies. Sedimentation velocity experiments implied that several of the phage DNAs were probably heavily glycosylated or methylated. These modifications might explain the partial or slow digestion of some of the DNAs by several of the 23 restriction enzymes tested. Protoplasts of the appropriate Streptomyces strains could be efficiently transfected with phage DNA in the presence of 25% (w/v) polyethylene glycol (mol. wt 6000).

  2. Visceral mobilization can lyse and prevent peritoneal adhesions in a rat model.

    Science.gov (United States)

    Bove, Geoffrey M; Chapelle, Susan L

    2012-01-01

    Peritoneal adhesions are almost ubiquitous following surgery. Peritoneal adhesions can lead to bowel obstruction, digestive problems, infertility, and pain, resulting in many hospital readmissions. Many approaches have been used to prevent or treat adhesions, but none offer reliable results. A method that consistently prevented or treated adhesions would benefit many patients. We hypothesized that an anatomically-based visceral mobilization, designed to promote normal mobility of the abdominal contents, could manually lyse and prevent surgically-induced adhesions. Cecal and abdominal wall abrasion was used to induce adhesions in 3 groups of 10 rats (Control, Lysis, and Preventive). All rats were evaluated 7 days following surgery. On postoperative day 7, unsedated rats in the Lysis group were treated using visceral mobilization, consisting of digital palpation, efforts to manually lyse restrictions, and mobilization of their abdominal walls and viscera. This was followed by immediate post-mortem adhesion evaluation. The rats in the Preventive group were treated daily in a similar fashion, starting the day after surgery. Adhesions in the Control rats were evaluated 7 days after surgery without any visceral mobilization. The therapist could palpate adhesions between the cecum and other viscera or the abdominal wall. Adhesion severity and number of adhesions were significantly lower in the Preventive group compared to other groups. In the Lysis and Preventive groups there were clear signs of disrupted adhesions. These initial observations support visceral mobilization may have a role in the prevention and treatment of post-operative adhesions. Copyright © 2011 Elsevier Ltd. All rights reserved.

  3. STUDIES ON THE BACTERIOPHAGE OF D'HERELLE : VI. ON THE VIRULENCE OF THE OVERGROWTH IN THE LYSED CULTURES OF BACILLUS PESTIS CAVIAE (M. T. II).

    Science.gov (United States)

    Bronfenbrenner, J; Muckenfuss, R S; Korb, C

    1926-10-31

    Resistants isolated from the overgrowth of cultures of B. pestis caviae (M. T. II) lysed by various strains of specific bacteriophage proved to be avirulent when administered to mice by feeding, or by intraperitoneal injection. These cultures remained resistant to the action of bacteriophage so long as they were carried on agar. When transferred to broth, however, one group of resistants, namely, those isolated by means of "weak" phages, became susceptible to lysis after five to seven daily passages. The other group of resistants, isolated from the cultures lysed by one of the "strong" phages, failed to become susceptible to lysis even after nearly 200 passages in broth. Simultaneously with the recovery of susceptibility, the cultures of the first group regained a degree of virulence comparable to that of the parent culture of B. pestis caviae. The cultures of the second group of resistants have failed thus far to recover virulence (10 months after isolation). The latter cultures, apart from lack of both virulence and susceptibility to lysis, are identical with the parent culture of B. pestis caviae, as indicated by biochemical and antigenic properties. Our findings offer evidence in favor of the view that resistant strains result from selection among variants already existing in the parent culture and do not arise through the inheritance of specific immunity properties produced by the action of phage.

  4. Wind characterization for design and comparison with standards, an example from Lyse at the Swedish west coast

    Energy Technology Data Exchange (ETDEWEB)

    Ganander, H. [Teknikgruppen AB, Sollentuna (Sweden); Carlen, I. [Chalmers Univ. of Technology, Goeteborg (Sweden). Div. of Building Technology; Bergstroem, H. [Uppsala Univ. (Sweden). Dept. of Meteorology

    1996-12-01

    The Lyse site at the Swedish west coast is an area with an archipelago of rocky islands to the west and an equally rocky mainland to the east. In between there are some open sea areas. As being the responsible project manager for the erection and the operation of a turbine at a site like Lyse, the question arises about characterization of the wind for design or purchase of a wind turbine. Or in other words what wind turbine class has to be used for the design, according to existing standards like for example IEC-1400 ? 3 refs, 10 figs

  5. Isolation,Identifcation and Characterization of Algae-lysing Strain H5 from Xiangxi Bay of Three Gorges Reservoir%溶藻细菌H5的分离、鉴定及溶藻特性研究

    Institute of Scientific and Technical Information of China (English)

    刘国勇; 胡亚平; 石小丹; 聂小倩; 黄应平

    2012-01-01

    [Objective] The study was conducted to isolate and identify alage-lysing bacterium and explore the characterization of alage-lysing bacterium so as to provide reference for the control of algae bloom. [ Method] A high-efficient algae-lysing strain H5 was isolated during spring blooms in Xiangxi Bay at Three Gorges Reservoir. According to the similarity analysis of its 16S rRNA gene sequence and by Biolog microsta-tion system, the strain was identified. By direct counting, the lytic efficiency of H5 to Stephanodiscus hantzschii, Peridiniopsis niei, Komma caudata and performing mode of H5 to S. hantzschii were studied. [ Result] 16S rDNA sequence analysis and Biolog analysis showed that H5 strain belonged to Lysinibacillus fusiformis. The highest and lowest algicidal rate of the strain was 71.3% and 57.4% respectively in the cultures of 5. hantzschii, P. niel, and K. caudata. The filtrate of cell spent media, and the heated above filtrate displayed the same algae-lytic ability, while the cell-free supernatant of the cells of bacteria showed no algae-lytic ability, indicating that some extracellular and thermo-stable substances were produced by this strain. [Conclusion] The strain H5 produced better lytic efficiency to S. hantzschii and performed algae-lytic ability by producing extracellular substances.%[目的]分离和鉴定溶藻细菌,研究其溶藻特性,为进一步研究溶藻细菌对水华的治理作用提供帮助.[方法]从香溪河春季水华集聚区水体中分离得到1株有高效溶藻效果的菌株(H5),采用16S rDNA序列相似性分析和Biolog微生物自动鉴定系统等对细菌进行鉴定.采用直接计数法,研究了其对汉斯冠盘藻(Stephanodiscus hantzschii、倪氏拟多甲藻(Peridiniopsis niei)、具尾逗隐藻(Kommia caudata)的抑制效果,及对汉斯冠盘藻的溶藻作用方式.[结果]根据生理生化及16S rDNA序列分析鉴定,H5属于纺缍形赖氨酸芽孢杆菌(Lysinibacillus fusiformis).该菌对汉斯冠

  6. Specific killing of P53 mutated tumor cell lines by a cross-reactive human HLA-A2-restricted P53-specific CTL line

    DEFF Research Database (Denmark)

    Würtzen, P A; Pedersen, L O; Poulsen, H S

    2001-01-01

    , 4 HLA-A2(+) p53-mutated tumor cell lines were lysed by the CTL line, indicating that these peptides are endogenously processed and presented on HLA-A2 molecule. Thus, monocyte-derived DC pulsed with a pool of peptides are able to induce CTL reactivity to wild-type p53 peptides presented by several...

  7. Certain Red Blood Cell Indices of Maternal and Umbilical Cord ...

    African Journals Online (AJOL)

    Uche

    umbilical cord packed cell volume and haemoglobin concentration in our locality. Keywords: Umbilical ... parasitaemia, or had premature delivery, history of haemorheological ... labour (immediately after delivery) by clamping and cutting the ...

  8. Roles of export genes cgmA and lysE for the production of L-arginine and L-citrulline by Corynebacterium glutamicum.

    Science.gov (United States)

    Lubitz, Dorit; Jorge, João M P; Pérez-García, Fernando; Taniguchi, Hironori; Wendisch, Volker F

    2016-10-01

    L-arginine is a semi-essential amino acid with application in cosmetic, pharmaceutical, and food industries. Metabolic engineering strategies have been applied for overproduction of L-arginine by Corynebacterium glutamicum. LysE was the only known L-arginine exporter of this bacterium. However, an L-arginine-producing strain carrying a deletion of lysE still accumulated about 10 mM L-arginine in the growth medium. Overexpression of the putative putrescine and cadaverine export permease gene cgmA was shown to compensate for the lack of lysE with regard to L-arginine export. Moreover, plasmid-borne overexpression of cgmA rescued the toxic effect caused by feeding of the dipeptide Arg-Ala to lysE-deficient C. glutamicum and argO-deficient Escherichia coli strains. Deletion of the repressor gene cgmR improved L-arginine titers by 5 %. Production of L-lysine and L-citrulline was not affected by cgmA overexpression. Taken together, CgmA may function as an export system not only for the diamine putrescine and cadaverine but also for L-arginine. The major export system for L-lysine and L-arginine LysE may also play a role in L-citrulline export since production of L-citrulline was reduced when lysE was deleted and improved by 45 % when lysE was overproduced.

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

    DEFF Research Database (Denmark)

    Rasmussen, E.S.

    1999-01-01

    The performance of two cell viability test kits based on the use of redox indicators yielding fluorescent products, the AlamarBlue assay and a resazurin-based in vitro toxicology assay kit from Sigma, was compared in the present study. Cultures of human neonatal foreskin fibroblasts were exposed...... components were tentatively identified as resazurin and resorufin. The AlamarBlue assay has gained wide application as a cell viability indicator that allows continuous monitoring of cell proliferation or cytotoxicity in human and animal cells, bacteria, and fungi, but no studies with the deliberate use...... of resazurin reduction to measure cell. proliferation in cultures of somatic mammalian cells have been published. In the AlamarBlue dye solution, resazurin is supplemented with various stabilizing agents, but the need for their use is questioned....

  10. [Characteristics of a new cyanophage lysing unicellular cyanobacteria of the genus Synechococcus].

    Science.gov (United States)

    Khudiakov, I Ia

    1977-01-01

    A new cyanophage S-2L growing on three strains of the cyanobacterium belonging to the Synechococcus genus has been isolated. The cyanophage has an icosahedral head, 56 nm in diameter, and flexible tail with a non-contracting sheath, 120 nm long. Over 95 per cent of the cyanophage particles are adsorbed within 10 min, the rate constant of adsorption being 3.2-10(-9) ml/min. The latent period lasts 5 hours, the yield is 100 particles per cell. The intracellular growth is characterized by the accumulation of the cyanophage particles at the poles of the cell. The photosynthetic lamellae remain intact up to the beginning of lysis which consists in local disruptions of the cell envelope.

  11. Naive Treg-like CCR7(+) mononuclear cells indicate unfavorable prognosis in hepatocellular carcinoma.

    Science.gov (United States)

    Shi, Jie-Yi; Duan, Meng; Sun, Qi-Man; Yang, Liuxiao; Wang, Zhi-Chao; Mynbaev, Ospan A; He, Yi-Feng; Wang, Ling-Yan; Zhou, Jian; Tang, Qi-Qun; Cao, Ya; Fan, Jia; Wang, Xiao-Ying; Gao, Qiang

    2016-07-01

    Chemokine receptor-like 1 (CCRL1) has the potential in creating a low level of CCL19 and CCL21 to hinder CCR7(+) cell tracking to tumor tissue. Previously, we found a tumor suppressive role of CCRL1 by impairing CCR7-related chemotaxis of tumor cells in human hepatocellular carcinoma (HCC). Here, we reported a contribution of CCR7(+) mononuclear cells in the tumor microenvironment to the progression of disease. Immunohistochemistry was used to investigate the distribution and clinical significance of CCR7(+) cells in a cohort of 240 HCC patients. Furthermore, the phenotype, composition, and functional status of CCR7(+) cells were determined by flow cytometry, immunofluorescence, and in vitro co-culture assays. We found that CCR7(+) mononuclear cells were dispersed around tumor tissue and negatively related to tumoral expression of CCRL1 (P CCR7(+) mononuclear cells positively correlated with the absence of tumor capsule, vascular invasion, and poor differentiation (P CCR7(+) mononuclear cells was significantly associated with worse survival and increased recurrence. We found that CCR7(+) mononuclear cells featured a naive Treg-like phenotype (CD45RA(+)CD25(+)FOXP3(+)) and possessed tumor-promoting potential by producing TGF-β1. Moreover, CCR7(+) cells were also composed of several immunocytes, a third of which were CD8(+) T cells. CCR7(+) Treg-like cells facilitate tumor growth and indicate unfavorable prognosis in HCC patients, but fortunately, their tracking to tumor tissue is under the control of CCRL1.

  12. The Effect of Predators on Cholera Biofilms: If it Lyses, We Can Smash It

    Science.gov (United States)

    Kalziqi, Arben; Bernardy, Eryn; Thomas, Jacob; Ratcliff, Will; Hammer, Brian; Yunker, Peter

    Many microbes form biofilms--dense clumps of cells and proteins--on surfaces. Biofilms are complex communities that facilitate the study of biological competition (e.g., two types of microbes may compete to form a biofilm in the same location) and interesting physics (e.g., the source of a biofilm's rigidity). Vibrio cholerae can produce biofilms which have a network-like structure--however, cholera can be genetically engineered to kill other cholera with different genotypes, which leaves behind a structureless ``slime'' rather than such a biofilm. Through mechanical creep testing of both predator-prey and non-predator populations, we found that the predator-prey population responds viscously and decreases in height with repeated compression, whereas the non-predator population responds elastically and maintains its original height. The current work suggests that cell lysis after killing disrupts biofilm formation, preventing microbial colonies from forming rigid networks.

  13. Real-time motion analysis reveals cell directionality as an indicator of breast cancer progression.

    Directory of Open Access Journals (Sweden)

    Michael C Weiger

    Full Text Available Cancer cells alter their migratory properties during tumor progression to invade surrounding tissues and metastasize to distant sites. However, it remains unclear how migratory behaviors differ between tumor cells of different malignancy and whether these migratory behaviors can be utilized to assess the malignant potential of tumor cells. Here, we analyzed the migratory behaviors of cell lines representing different stages of breast cancer progression using conventional migration assays or time-lapse imaging and particle image velocimetry (PIV to capture migration dynamics. We find that the number of migrating cells in transwell assays, and the distance and speed of migration in unconstrained 2D assays, show no correlation with malignant potential. However, the directionality of cell motion during 2D migration nicely distinguishes benign and tumorigenic cell lines, with tumorigenic cell lines harboring less directed, more random motion. Furthermore, the migratory behaviors of epithelial sheets observed under basal conditions and in response to stimulation with epidermal growth factor (EGF or lysophosphatitic acid (LPA are distinct for each cell line with regard to cell speed, directionality, and spatiotemporal motion patterns. Surprisingly, treatment with LPA promotes a more cohesive, directional sheet movement in lung colony forming MCF10CA1a cells compared to basal conditions or EGF stimulation, implying that the LPA signaling pathway may alter the invasive potential of MCF10CA1a cells. Together, our findings identify cell directionality as a promising indicator for assessing the tumorigenic potential of breast cancer cell lines and show that LPA induces more cohesive motility in a subset of metastatic breast cancer cells.

  14. Characterization of novel bacteriophage phiC119 capable of lysing multidrug-resistant Shiga toxin-producing Escherichia coli O157:H7

    Science.gov (United States)

    Amarillas, Luis; Chaidez, Cristóbal; González-Robles, Arturo; Lugo-Melchor, Yadira

    2016-01-01

    Background Shiga toxin-producing Escherichia coli (STEC) is one of the most common and widely distributed foodborne pathogens that has been frequently implicated in gastrointestinal and urinary tract infections. Moreover, high rates of multiple antibiotic-resistant E. coli strains have been reported worldwide. Due to the emergence of antibiotic-resistant strains, bacteriophages are considered an attractive alternative to biocontrol pathogenic bacteria. Characterization is a preliminary step towards designing a phage for biocontrol. Methods In this study, we describe the characterization of a bacteriophage designated phiC119, which can infect and lyse several multidrug-resistant STEC strains and some Salmonella strains. The phage genome was screened to detect the stx-genes using PCR, morphological analysis, host range was determined, and genome sequencing were carried out, as well as an analysis of the cohesive ends and identification of the type of genetic material through enzymatic digestion of the genome. Results Analysis of the bacteriophage particles by transmission electron microscopy showed that it had an icosahedral head and a long tail, characteristic of the family Siphoviridae. The phage exhibits broad host range against multidrug-resistant and highly virulent E. coli isolates. One-step growth experiments revealed that the phiC119 phage presented a large burst size (210 PFU/cell) and a latent period of 20 min. Based on genomic analysis, the phage contains a linear double-stranded DNA genome with a size of 47,319 bp. The phage encodes 75 putative proteins, but lysogeny and virulence genes were not found in the phiC119 genome. Conclusion These results suggest that phage phiC119 may be a good biological control agent. However, further studies are required to ensure its control of STEC and to confirm the safety of phage use. PMID:27672499

  15. Characterization of novel bacteriophage phiC119 capable of lysing multidrug-resistant Shiga toxin-producing Escherichia coli O157:H7

    Directory of Open Access Journals (Sweden)

    Luis Amarillas

    2016-09-01

    Full Text Available Background Shiga toxin-producing Escherichia coli (STEC is one of the most common and widely distributed foodborne pathogens that has been frequently implicated in gastrointestinal and urinary tract infections. Moreover, high rates of multiple antibiotic-resistant E. coli strains have been reported worldwide. Due to the emergence of antibiotic-resistant strains, bacteriophages are considered an attractive alternative to biocontrol pathogenic bacteria. Characterization is a preliminary step towards designing a phage for biocontrol. Methods In this study, we describe the characterization of a bacteriophage designated phiC119, which can infect and lyse several multidrug-resistant STEC strains and some Salmonella strains. The phage genome was screened to detect the stx-genes using PCR, morphological analysis, host range was determined, and genome sequencing were carried out, as well as an analysis of the cohesive ends and identification of the type of genetic material through enzymatic digestion of the genome. Results Analysis of the bacteriophage particles by transmission electron microscopy showed that it had an icosahedral head and a long tail, characteristic of the family Siphoviridae. The phage exhibits broad host range against multidrug-resistant and highly virulent E. coli isolates. One-step growth experiments revealed that the phiC119 phage presented a large burst size (210 PFU/cell and a latent period of 20 min. Based on genomic analysis, the phage contains a linear double-stranded DNA genome with a size of 47,319 bp. The phage encodes 75 putative proteins, but lysogeny and virulence genes were not found in the phiC119 genome. Conclusion These results suggest that phage phiC119 may be a good biological control agent. However, further studies are required to ensure its control of STEC and to confirm the safety of phage use.

  16. Musashi1 regulates breast tumor cell proliferation and is a prognostic indicator of poor survival

    Directory of Open Access Journals (Sweden)

    Wang Xiao-Yang

    2010-08-01

    Full Text Available Abstract Background Musashi1 (Msi1 is a conserved RNA-binding protein that regulates the Notch and Wnt pathways, and serves as a stem cell marker in the breast and other tissues. It is unknown how Msi1 relates to other breast cancer markers, whether it denotes tumor initiating cells (TICs, and how it affects gene expression and tumor cell survival in breast cancer cells. Results Msi1 expression was analyzed in 20 breast cancer cell lines and in 140 primary breast tumors by western blotting and immunohistochemistry, respectively. Lentivirus RNA interference was used to reduce Msi1 expression in breast cancer cell lines MCF-7 and T47D grown as spheroid cultures and to assess stem cell gene expression and the growth of these cell lines as xenografts. In normal human breast tissue, Msi1 was expressed in 10.6% of myoepithelum and 1.2% of ductal epithelium in the terminal ductal lobular unit (TDLU, whereas, less than 0.05% of ductal epithelium and myoepithelium in large ducts outside the TDLU expressed Msi1. Msi1 was expressed in 55% of the breast cancer cell lines and correlated with ErbB2 expression in 50% of the cell lines. Msi1 was expressed in 68% of primary tumors and in 100% of lymph node metastases, and correlated with 5 year survival. Msi1 was enriched in CD133+ MCF-7 and T47D cells and in spheroid cultures of these cells, and Msi1 'knockdown' (KD with a lentivirus-expressed shRNA decreased the number and size of spheroid colonies. Msi1 KD reduced Notch1, c-Myc, ErbB2 and pERK1/2 expression, and increased p21CIP1 expression, which is consistent with known Msi1 target mRNAs. Msi1 KD also reduced the expression of the somatic and embryonic stem cell markers, CD133, Bmi1, Sox2, Nanog and Oct4. Xenografts of MCF-7 and T47D Msi1 KD cells resulted in a marked reduction of tumor growth, reduced Msi1 and Notch1 expression and increased p21CIP1 expression. Conclusion Msi1 is a negative prognostic indicator of breast cancer patient survival, and is

  17. Parathyroid hormone-related protein serves as a prognostic indicator in oral squamous cell carcinoma.

    Science.gov (United States)

    Lv, Zhongjing; Wu, Xiangbing; Cao, Wei; Shen, ZongZe; Wang, Lizhen; Xie, FuRong; Zhang, JianJun; Ji, Tong; Yan, Ming; Chen, WanTao

    2014-12-18

    In our previous study, parathyroid hormone-like hormone (PTHLH) which encodes parathyroid hormone-related protein (PTHrP) was revealed to be up-regulated in oral squamous cell carcinoma (OSCC) compared with paired apparently normal surgical margins using microarray method. However, the function and prognostic indicators of PTHLH/PTHrP in OSCC remain obscure. The mRNA levels of PTHLH and its protein levels were investigated in 9 OSCC cell lines and in 36 paired OSCC specimens by real-time PCR and western blotting. The biological function of PTHLH/PTHrP was investigated using small interfering RNA (siRNA) in 3 OSCC cell lines, and immunohistochemistry was used to estimate the prognostic value of PTHrP in 101 patients with head and neck squamous cell carcinoma (HNSCC), including OSCC and oropharyngeal squamous cell carcinoma. Cell cycle was tested by flow cytometry and cell cycle related genes were investigated by western blotting and immunocytochemistry assay. This study showed that the mRNA and protein levels of PTHLH in 9 OSCC cell lines were much higher than that in normal epithelial cells (P PTHrP by siRNAs could reduce cell proliferation and inhibit plate and soft agar colony formation as well as affect the cell cycle of OSCC cells. The key proteins related to the cell cycle were changed by anti-PTHLH siRNA. The results showed that cyclin D1 and CDK4 expressions were significantly reduced in the cells transfected with anti-PTHLH siRNA. On the other hand, the expression of p21 was increased. The results also showed that high PTHrP level was associated with poor pathologic differentiation (P = 0.0001) and poor prognosis (P = 0.0003) in patients with HNSCC. This study suggests that PTHLH/PTHrP is up-regulated in OSCCs. Therefore, PTHLH/PTHrP could play a role in the pathogenesis of OSCC by affecting cell proliferation and cell cycle, and the protein levels of PTHrP might serve as a prognostic indicator for evaluating patients with HNSCCs.

  18. An Efficient DNA Extraction Method for Lactobacillus casei, a Difficult-to-Lyse Bacterium

    Directory of Open Access Journals (Sweden)

    Alimolaei

    2016-02-01

    Full Text Available Background There are several protocols to extract DNA from Lactobacillus spp. In the case of L. casei it is harder because of its especial and thick cell wall. Objectives In this study, nine DNA extraction protocols (by lysozyme treatment were evaluated and compared in two categories (traditional and kit-based protocols and an improved method was presented. Materials and Methods DNA quantity and quality was determined by spectrophotometry, agarose gel electrophoresis and polymerase chain reaction (PCR. Results The results revealed that the yield of extracted DNA differed by each protocol (5.8 - 17.1 μg/100 μL, but provided appropriate DNA for PCR amplification. The modified protocol offered the best total DNA extraction method when both quality (DNA purity; 1.54 μg and quantity (DNA yield; 17.1 μg were considered. Conclusions We suggest this protocol for effective and inexpensive DNA isolation from L. casei for downstream biological processes such as PCR.

  19. Bioluminescent Indicator for Highly Sensitive Analysis of Estrogenic Activity in a Cell-Based Format.

    Science.gov (United States)

    Takenouchi, Osamu; Kanno, Akira; Takakura, Hideo; Hattori, Mitsuru; Ozawa, Takeaki

    2016-11-16

    Estrogens regulate different physiological systems with wide ranges of concentrations. The rapid analysis of estrogens is crucially important for drug discovery and medical diagnosis, but quantitation of nanomolar estrogens in live cells persists as an important challenge. We herein describe a bioluminescent indicator used to detect low concentrations of estrogens quantitatively with a high signal-to-background ratio. The indicator comprises a ligand-binding domain of an estrogen receptor connected with its binding peptide, which is sandwiched between split fragments of a luciferase mutant. Results show that the indicator recovered its bioluminescence upon binding to 17β-estradiol at concentrations higher than 1.0 × 10(-10) M. The indicator was reactive to agonists but did not respond to antagonists. The indicator is expected to be applicable for rapid screening estrogenic compounds and inhibitors, facilitating the discovery of drug candidates in a high-throughput manner.

  20. Indications and Results of HLA-Identical Sibling Hematopoietic Cell Transplantation for Sickle Cell Disease.

    Science.gov (United States)

    Walters, Mark C; De Castro, Laura M; Sullivan, Keith M; Krishnamurti, Lakshmanan; Kamani, Naynesh; Bredeson, Christopher; Neuberg, Donna; Hassell, Kathryn L; Farnia, Stephanie; Campbell, Andrew; Petersdorf, Effie

    2016-02-01

    Although a number of published trials exist of HLA-identical sibling hematopoietic cell transplantation (HCT) for sickle cell disease (SCD) that span 2 decades, when and for whom this therapy should be pursued is a subject of debate. Assessments of the risks of transplant-related complications that include infertility and debilitating graft-versus-host disease and long-term quality of life after successful HCT are difficult to perform without prospective trials in transplant and nontransplant cohorts. However, it is possible to assess the risk of mortality and to compare published rates of survival in individuals with SCD treated and not treated by HCT. In this brief review, projections about mortality risk based on recent published reports are reviewed and summarized. The published data show overall survival and event-free survival rates of 95% and 92%, respectively, in children treated by HLA-identical sibling HCT. The overall survival rates in the Center for International Blood and Marrow Transplant Research (N = 412) and European Blood and Marrow Transplant (N = 487) registries were 91% and 95%, respectively. These results provide broad support for the therapeutic value of HLA-identical sibling HCT for children with SCD and serve as the basis for a strong recommendation in favor of the option of HCT when a suitable donor is available. The experience of HLA-identical sibling HCT in adults with SCD is limited but appears to be similar to results in children. These preliminary observations, however, warrant further investigation.

  1. Chromatin organization as an indicator of glucocorticoid induced natural killer cell dysfunction.

    Science.gov (United States)

    Misale, Michael S; Witek Janusek, Linda; Tell, Dina; Mathews, Herbert L

    2017-09-12

    It is well-established that psychological distress reduces natural killer cell immune function and that this reduction can be due to the stress-induced release of glucocorticoids. Glucocorticoids are known to alter epigenetic marks associated with immune effector loci, and are also known to influence chromatin organization. The purpose of this investigation was to assess the effect of glucocorticoids on natural killer cell chromatin organization and to determine the relationship of chromatin organization to natural killer cell effector function, e.g. interferon gamma production. Interferon gamma production is the prototypic cytokine produced by natural killer cells and is known to modulate both innate and adaptive immunity. Glucocorticoid treatment of human peripheral blood mononuclear cells resulted in a significant reduction in interferon gamma production. Glucocorticoid treatment also resulted in a demonstrable natural killer cell nuclear phenotype. This phenotype was localization of the histone, post-translational epigenetic mark, H3K27me3, to the nuclear periphery. Peripheral nuclear localization of H3K27me3 was directly related to cellular levels of interferon gamma. This nuclear phenotype was determined by direct visual inspection and by use of an automated, high through-put technology, the Amnis ImageStream. This technology combines the per-cell information content provided by standard microscopy with the statistical significance afforded by large sample sizes common to standard flow cytometry. Most importantly, this technology provides for a direct assessment of the localization of signal intensity within individual cells. The results demonstrate glucocorticoids to dysregulate natural killer cell function at least in part through altered H3K27me3 nuclear organization and demonstrate H3K27me3 chromatin organization to be a predictive indicator of glucocorticoid induced immune dysregulation of natural killer cells. Copyright © 2017 Elsevier Inc. All rights

  2. Single cell genomics indicates horizontal gene transfer and viral infections in a deep subsurface Firmicutes population

    Directory of Open Access Journals (Sweden)

    Jessica eLabonté

    2015-04-01

    Full Text Available A major fraction of Earth's prokaryotic biomass dwells in the deep subsurface, where cellular abundances per volume of sample are lower, metabolism is slower, and generation times are longer than those in surface terrestrial and marine environments. How these conditions impact biotic interactions and evolutionary processes is largely unknown. Here we employed single cell genomics to analyze cell-to-cell genome content variability and signatures of horizontal gene transfer (HGT and viral infections in five cells of Candidatus Desulforudis audaxviator, which were collected from a three km-deep fracture water in the 2.9 Ga-old Witwatersrand Basin of South Africa. Between 0 and 32 % of genes recovered from single cells were not present in the original, metagenomic assembly of Desulforudis, which was obtained from a neighboring subsurface fracture. We found a transposable prophage, a retron, multiple clustered regularly interspaced short palindromic repeats (CRISPRs and restriction-modification systems, and an unusually high frequency of transposases in the analyzed single cell genomes. This indicates that recombination, HGT and viral infections are prevalent evolutionary events in the studied population of microorganisms inhabiting a highly stable deep subsurface environment.

  3. Intracellular pH distribution as a cell health indicator in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Aabo, Thomas; Glückstad, Jesper; Siegumfeldt, Henrik

    2011-01-01

    Internal pH regulation is vital for many cell functions, including transport mechanisms and metabolic enzyme activity. More specifically, transport mechanisms are to a wide degree governed by internal pH distributions. We introduce the term standard deviation of the intracellular pH (s.......d.(pHint)) to describe the internal pH distributions. The cellular pH distributional response to external stress such as heat has not previously been determined. In this study, the intracellular pH (pHi) and the s.d.(pHint) of Saccharomyces cerevisiae cells exposed to supralethal temperatures were measured using...... temperatures than pHi. s.d.(pHint) is therefore proposed as an early health/vitality indicator in S. cerevisiae cells exposed to heat stress....

  4. Microcystins derived from lysing Microcystis cells do not cause negative effects on crustacean zooplankton in Lake Taihu, China

    NARCIS (Netherlands)

    Chen, F.; Dai, X.; Shu, T.; Gulati, R.D.; Liu, Z.

    2013-01-01

    Microcystins (MCs) have a toxic effect on crustacean zooplankton in the laboratory, but there is little or no unequivocal evidence in the literature of their lethal effects on crustacean zooplankton in the field. We used the natural microcystins extracted from Microcystis spp. to test if they could

  5. A comparison of fluorescent Ca2+ indicators for imaging local Ca2+ signals in cultured cells

    Science.gov (United States)

    Lock, Jeffrey T.; Parker, Ian

    2015-01-01

    Localized subcellular changes in Ca2+ serve as important cellular signaling elements, regulating processes as diverse as neuronal excitability and gene expression. Studies of cellular Ca2+ signaling have been greatly facilitated by the availability of fluorescent Ca2+ indicators. The respective merits of different indicators to monitor bulk changes in cellular Ca2+ levels have been widely evaluated, but a comprehensive comparison for their use in detecting and analyzing local, subcellular Ca2+ signals is lacking. Here, we evaluated several fluorescent Ca2+ indicators in the context of local Ca2+ signals (puffs) evoked by inositol 1,4,5-trisphosphate (IP3) in cultured human neuroblastoma SH-SY5Y cells, using high-speed video-microscopy. Altogether, nine synthetic Ca2+ dyes (Fluo-4, Fluo-8, Fluo-8 high affinity, Fluo-8 low affinity, Oregon Green BAPTA-1, Cal-520, Rhod-4, Asante Calcium Red, and X-Rhod-1) and three genetically-encoded Ca2+-indicators (GCaMP6-slow, -medium and -fast variants) were tested; criteria include the magnitude, kinetics, signal-to-noise ratio and detection efficiency of local Ca2+ puffs. Among these, we conclude that Cal-520 is the optimal indicator for detecting and faithfully tracking local events; that Rhod-4 is the red-emitting indicator of choice; and that none of the GCaMP6 variants are well suited for imaging subcellular Ca2+ signals. PMID:26572560

  6. In HepG2 cells, coexisting carnitine deficiency masks important indicators of marginal biotin deficiency.

    Science.gov (United States)

    Bogusiewicz, Anna; Boysen, Gunnar; Mock, Donald M

    2015-01-01

    A large number of birth defects are related to nutrient deficiencies; concern that biotin deficiency is teratogenic in humans is reasonable. Surprisingly, studies indicate that increased urinary 3-hydroxyisovalerylcarnitine (3HIAc), a previously validated marker of biotin deficiency, is not a valid biomarker in pregnancy. In this study we hypothesized that coexisting carnitine deficiency can prevent the increase in 3HIAc due to biotin deficiency. We used a 2-factor nutrient depletion design to induce isolated and combined biotin and carnitine deficiency in HepG2 cells and then repleted cells with carnitine. To elucidate the metabolic pathogenesis, we quantitated intracellular and extracellular free carnitine, acylcarnitines, and acylcarnitine ratios using liquid chromatography-tandem mass spectrometry. Relative to biotin-sufficient, carnitine-sufficient cells, intracellular acetylcarnitine increased by 90%, propionylcarnitine more than doubled, and 3HIAc increased by >10-fold in biotin-deficient, carnitine-sufficient (BDCS) cells, consistent with a defensive mechanism in which biotin-deficient cells transesterify the acyl-coenzyme A (acyl-CoA) substrates of the biotin-dependent carboxylases to the related acylcarnitines. Likewise, in BDCS cells, the ratio of acetylcarnitine to malonylcarnitine and the ratio of propionylcarnitine to methylmalonylcarnitine both more than tripled, and the ratio of 3HIAc to 3-methylglutarylcarnitine (MGc) increased by >10-fold. In biotin-deficient, carnitine-deficient (BDCD) cells, the 3 substrate-derived acylcarnitines changed little, but the substrate:product ratios were masked to a lesser extent. Moreover, carnitine repletion unmasked biotin deficiency in BDCD cells as shown by increases in acetylcarnitine, propionylcarnitine, and 3HIAc (each increased by >50-fold). Likewise, ratios of acetylcarnitine:malonylcarnitine, propionylcarnitine:methylmalonylcarnitine, and 3HIAc:MGc all increased by >8-fold. Our findings provide strong

  7. Effect of artificial photoperiod on the blood cell indices of the catfish, Clarias batrachus

    Directory of Open Access Journals (Sweden)

    S Srivastava

    2010-04-01

    Full Text Available The present study is aimed to assess the influence of artificial photoperiod on the blood cell indices of an Indian catfish, Clarias batrachus(Linn.. Blood samples taken from adult fishes exposed to artificial photoperiod of 24L:0D and 0L:24D for a short period of 24 hrs, were analyzed for total RBC, total WBC, differential leukocyte count (DLC and some physiological variables – glucose and chloride. The total RBC and WBC counts were unaffected by both the artificial photoperiod regimes. However, lymphopenia (p<.05 and neutrophilia (p<.05 were observed under 24L:0D photoperiod. Blood chloride levels were significantly higher (p<.05 in 24L:0D whereas blood glucose levels remained unchanged in both the photoperiod treatments. The 0L:24D photoperiod did not produce any significant change in the blood cell indices neither in the physiological variables. The findings indicate that exposure to continuous light for as short a duration as of 24 hrs elicits stress responses in the leukocyte profile of this nocturnal fish and highlights the role of leukocyte profile as potential stress biomarkers in vertebrates.

  8. High virus-to-cell ratios indicate ongoing production of viruses in deep subsurface sediments.

    Science.gov (United States)

    Engelhardt, Tim; Kallmeyer, Jens; Cypionka, Heribert; Engelen, Bert

    2014-07-01

    Marine sediments cover two-thirds of our planet and harbor huge numbers of living prokaryotes. Long-term survival of indigenous microorganisms within the deep subsurface is still enigmatic, as sources of organic carbon are vanishingly small. To better understand controlling factors of microbial life, we have analyzed viral abundance within a comprehensive set of globally distributed subsurface sediments. Phages were detected by electron microscopy in deep (320 m below seafloor), ancient (∼14 Ma old) and the most oligotrophic subsurface sediments of the world's oceans (South Pacific Gyre (SPG)). The numbers of viruses (10(4)-10(9) cm(-3), counted by epifluorescence microscopy) generally decreased with sediment depth, but always exceeded the total cell counts. The enormous numbers of viruses indicate their impact as a controlling factor for prokaryotic mortality in the marine deep biosphere. The virus-to-cell ratios increased in deeper and more oligotrophic layers, exhibiting values of up to 225 in the deep subsurface of the SPG. High numbers of phages might be due to absorption onto the sediment matrix and a diminished degradation by exoenzymes. However, even in the oldest sediments, microbial communities are capable of maintaining viral populations, indicating an ongoing viral production and thus, viruses provide an independent indicator for microbial life in the marine deep biosphere.

  9. Increased Expression of SETD7 Promotes Cell Proliferation by Regulating Cell Cycle and Indicates Poor Prognosis in Hepatocellular Carcinoma.

    Directory of Open Access Journals (Sweden)

    Yuanyuan Chen

    Full Text Available To investigate the role of SET domain containing 7 (SETD7 in hepatocellular carcinoma (HCC and determine whether SETD7 can be used as a predictor of overall survival in HCC patients.mRNAs and proteins of SETD7 and related genes in HCC tumor samples and paired adjacent non-tumorous liver tissues (ANLTs (n = 20 or culture cells were determined by quantitative real-time PCR and Western blot. Cell proliferation and apoptosis with SETD7 knockdown SMMC-7721 cells or SETD7 overexpressed HepG2 cells were analyzed by CCK8 assay or flow cytometry. Gene expression alterations in SETD7 knockdown of SMMC-7721 cells were determined by digital gene expression (DGE profiling. Defined data on patients (n = 225 with HCC were retrieved for the further study. Tissue microarrays (TMAs were performed using paraffin tissues with tumor and ANLTs. SETD7 and related proteins were determined by TMAs immunohistochemistry. Statistical analyses were conducted to associate SETD7 expression with tumor features and patient outcomes, as well as related proteins expression.SETD7 expression was significantly higher in HCC tumor tissues than in ANLTs. SETD7 overexpression in vitro can promote HepG2 cell proliferation, whereas SETD7 knockdown can inhibit SMMC-7721 cell proliferation by regulating the cell cycle. SETD7 expression was significantly correlated with five genes expression. Increased SETD7 is associated with metastasis, recurrence, large tumor size, and poor tumor differentiation, and indicates poor prognosis in HCC patients.SETD7 plays a critical role in HCC, and its immunohistochemistry signature provides potential clinical significance for personalized prediction of HCC prognosis.

  10. Erythropoietin reduces storage lesions and decreases apoptosis indices in blood bank red blood cells

    Science.gov (United States)

    Penuela, Oscar Andrés; Palomino, Fernando; Gómez, Lina Andrea

    2015-01-01

    Background Recent evidence shows a selective destruction of the youngest circulating red blood cells (neocytolysis) trigged by a drop in erythropoietin levels. Objective The aim of this study was to evaluate the effect of recombinant human erythropoietin beta on the red blood cell storage lesion and apoptosis indices under blood bank conditions. Methods Each one of ten red blood cell units preserved in additive solution 5 was divided in two volumes of 100 mL and assigned to one of two groups: erythropoietin (addition of 665 IU of recombinant human erythropoietin) and control (isotonic buffer solution was added). The pharmacokinetic parameters of erythropoietin were estimated and the following parameters were measured weekly, for six weeks: Immunoreactive erythropoietin, hemolysis, percentage of non-discocytes, adenosine triphosphate, glucose, lactate, lactate dehydrogenase, and annexin-V/esterase activity. The t-test or Wilcoxon's test was used for statistical analysis with significance being set for a p-value 6 weeks under blood bank conditions, with persistent supernatant concentrations of erythropoietin during the entire storage period. Adenosine triphosphate was higher in the Erythropoietin Group in Week 6 (4.19 ± 0.05 μmol/L vs. 3.53 ± 0.02 μmol/L; p-value = 0.009). The number of viable cells in the Erythropoietin Group was higher than in the Control Group (77% ± 3.8% vs. 71% ± 2.3%; p-value <0.05), while the number of apoptotic cells was lower (9.4% ± 0.3% vs. 22% ± 0.8%; p-value <0.05). Conclusions Under standard blood bank conditions, an important proportion of red blood cells satisfy the criteria of apoptosis. Recombinant human erythropoietin beta seems to improve storage lesion parameters and mitigate apoptosis. PMID:26969770

  11. Visualisation of cell cycle modifications by X-ray irradiation of single HeLa cells using fluorescent ubiquitination-based cell cycle indicators.

    Science.gov (United States)

    Kaminaga, K; Noguchi, M; Narita, A; Sakamoto, Y; Kanari, Y; Yokoya, A

    2015-09-01

    To explore the effects of X-ray irradiation on mammalian cell cycle dynamics, single cells using the fluorescent ubiquitination-based cell cycle indicator (Fucci) technique were tracked. HeLa cells expressing Fucci were used to visualise cell cycle modifications induced by irradiation. After cultured HeLa-Fucci cells were exposed to 5 Gy X-rays, fluorescent cell images were captured every 20 min for 48 h using a fluorescent microscope. Time dependence of the fluorescence intensity of S/G2 cells was analysed to examine the cell cycle dynamics of irradiated and non-irradiated control cells. The results showed that irradiated cells could be divided into two populations: one with similar cell cycle dynamics to that of non-irradiated cells, and another displaying a prolonged G2 phase. Based on these findings, it is proposed in this article that an underlying switch mechanism is involved in cell cycle regulation and the G2/M checkpoint of HeLa cells.

  12. Science Letters: Brain natriuretic peptide: A potential indicator of cardiomyogenesis after autologous mesenchymal stem cell transplantation?

    Institute of Scientific and Technical Information of China (English)

    LI Nan; WANG Jian-an

    2006-01-01

    We observed in a pilot study that there was a transient elevation of brain natriuretic peptide (BNP) level shortly after the transplantation in the patient with ischemic heart failure, which is unexplainable by the simultaneous increase of the cardiac output and six-minute walk distance. Similar findings were observed in the phase I trial. We postulated on the basis of the finding of Fukuda in vitro that this transient elevation of BNP level against the improvement of cardiac function and exercise capacity might indicate cardiomyogenesis in patients after mesenchymal stem cell transplantation. Further study is warranted to verify the hypothesis.

  13. Use of genetically-encoded calcium indicators for live cell calcium imaging and localization in virus-infected cells.

    Science.gov (United States)

    Perry, Jacob L; Ramachandran, Nina K; Utama, Budi; Hyser, Joseph M

    2015-11-15

    Calcium signaling is a ubiquitous and versatile process involved in nearly every cellular process, and exploitation of host calcium signals is a common strategy used by viruses to facilitate replication and cause disease. Small molecule fluorescent calcium dyes have been used by many to examine changes in host cell calcium signaling and calcium channel activation during virus infections, but disadvantages of these dyes, including poor loading and poor long-term retention, complicate analysis of calcium imaging in virus-infected cells due to changes in cell physiology and membrane integrity. The recent expansion of genetically-encoded calcium indicators (GECIs), including blue and red-shifted color variants and variants with calcium affinities appropriate for calcium storage organelles like the endoplasmic reticulum (ER), make the use of GECIs an attractive alternative for calcium imaging in the context of virus infections. Here we describe the development and testing of cell lines stably expressing both green cytoplasmic (GCaMP5G and GCaMP6s) and red ER-targeted (RCEPIAer) GECIs. Using three viruses (rotavirus, poliovirus and respiratory syncytial virus) previously shown to disrupt host calcium homeostasis, we show the GECI cell lines can be used to detect simultaneous cytoplasmic and ER calcium signals. Further, we demonstrate the GECI expression has sufficient stability to enable long-term confocal imaging of both cytoplasmic and ER calcium during the course of virus infections.

  14. Transient Oral Human Cytomegalovirus Infections Indicate Inefficient Viral Spread from Very Few Initially Infected Cells.

    Science.gov (United States)

    Mayer, Bryan T; Krantz, Elizabeth M; Swan, David; Ferrenberg, James; Simmons, Karen; Selke, Stacy; Huang, Meei-Li; Casper, Corey; Corey, Lawrence; Wald, Anna; Schiffer, Joshua T; Gantt, Soren

    2017-06-15

    Cytomegalovirus (CMV) is acquired by the oral route in children, and primary infection is associated with abundant mucosal replication, as well as the establishment of latency in myeloid cells that results in lifelong infection. The efficiency of primary CMV infection in humans following oral exposure, however, is unknown. We consistently detected self-limited, low-level oral CMV shedding events, which we termed transient CMV infections, in a prospective birth cohort of 30 highly exposed CMV-uninfected infants. We estimated the likelihood of transient oral CMV infections by comparing their observed frequency to that of established primary infections, characterized by persistent high-level shedding, viremia, and seroconversion. We developed mathematical models of viral dynamics upon initial oral CMV infection and validated them using clinical shedding data. Transient infections comprised 76 to 88% of oral CMV shedding events. For this high percentage of transient infections to occur, we identified two mathematical prerequisites: a very small number of initially infected oral cells (1 to 4) and low viral infectivity (<1.5 new cells infected/cell). These observations indicate that oral CMV infection in infants typically begins with a single virus that spreads inefficiently to neighboring cells. Thus, although the incidence of CMV infection is high during infancy, our data provide a mechanistic framework to explain why multiple CMV exposures are typically required before infection is successfully established. These findings imply that a sufficiently primed immune response could prevent CMV from establishing latent infection in humans and support the achievability of a prophylactic CMV vaccine.IMPORTANCE CMV infects the majority of the world's population and is a major cause of birth defects. Developing a vaccine to prevent CMV infection would be extremely valuable but would be facilitated by a better understanding of how natural human CMV infection is acquired. We

  15. Cocoa Extract Indicated Has Activity on Selectively Killing Breast Cancer Cells

    Directory of Open Access Journals (Sweden)

    Ariza Budi Tunjung Sari

    2015-09-01

    Full Text Available Effect of the cocoa crude extract on mortality of breast cancer cell lines i.e. MCF-7, T47D and normal cell (Vero, was observed. Crude cocoa extract prepared from a freshly dried cocoa bean that was containing 14% catechin and 0.6% caffeine. Catechin and caffeine content were modulated to 2-folds (28% catechin or 1.2% caffeine and 3-folds (42% catechin or 1.8% caffeine by adding pure compounds. Extracts were dissolved in dimethylsulfoxide (DMSO at concentrations ranging from 200 to 1600 μg/ml. The positive control was doxorubicin (0.5-16 μg/ml in DMSO. Cell lines (MCF-7, T47D, and Vero were incubated in test sample for 24h at 37°, prior to 3-(4,4-dimetylthiazole-2-yl-2,5-diphenyltetrazolium bromide (MTT assay. The absorbance of each well was measured at 550 nm, and lethal concentration (LC50 was calculated. The cocoa extract induced mortality of breast cancer cell lines but not in Vero cells. The effect on MCF-7 was greater than on T47D, given the LC50 was 1236 μg/ml (MCF-7 and 1893 μg/ml (T47D. Cytotoxic potential of cocoa extract was much lower than doxorubicin whose LC50 was 0,777 μg/ml (MCF-7 and 0,082 μg/ml (T47D. Increasing catechin content to 2-folds did not significantly affect LC50 value, but 3-folds catechin content reduced LC50 to 1021 μg/ml. Meanwhile increasing caffeine content to 2-folds significantly reduced LC50 to 750 μg/ml, however, 3-fold content resulted in slightly higher LC50 at 780 μg/ml. This indicates that cocoa extract have anti-cancer potential, and purification may improve this property.

  16. A Clinical Indications Prediction Scale Based on TWIST1 for Human Mesenchymal Stem Cells

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    Siddaraju V. Boregowda

    2016-02-01

    Full Text Available In addition to their stem/progenitor properties, mesenchymal stem cells (MSCs also exhibit potent effector (angiogenic, antiinflammatory, immuno-modulatory functions that are largely paracrine in nature. It is widely believed that effector functions underlie most of the therapeutic potential of MSCs and are independent of their stem/progenitor properties. Here we demonstrate that stem/progenitor and effector functions are coordinately regulated at the cellular level by the transcription factor Twist1 and specified within populations according to a hierarchical model. We further show that manipulation of Twist1 levels by genetic approaches or by exposure to widely used culture supplements including fibroblast growth factor 2 (Ffg2 and interferon gamma (IFN-gamma alters MSC efficacy in cell-based and in vivo assays in a predictable manner. Thus, by mechanistically linking stem/progenitor and effector functions our studies provide a unifying framework in the form of an MSC hierarchy that models the functional complexity of populations. Using this framework, we developed a CLinical Indications Prediction (CLIP scale that predicts how donor-to-donor heterogeneity and culture conditions impact the therapeutic efficacy of MSC populations for different disease indications.

  17. Visualizing cell-cycle kinetics after hypoxia/reoxygenation in HeLa cells expressing fluorescent ubiquitination-based cell cycle indicator (Fucci).

    Science.gov (United States)

    Goto, Tatsuaki; Kaida, Atsushi; Miura, Masahiko

    2015-12-10

    Hypoxia induces G1 arrest in many cancer cell types. Tumor cells are often exposed to hypoxia/reoxygenation, especially under acute hypoxic conditions in vivo. In this study, we investigated cell-cycle kinetics and clonogenic survival after hypoxia/reoxygenation in HeLa cells expressing fluorescent ubiquitination-based cell cycle indicator (Fucci). Hypoxic treatment halted cell-cycle progression during mid-S to G2 phase, as determined by the cell cycle-regulated E3 ligase activities of SCF(Skp2) and APC/C(Cdh1), which are regulators of the Fucci probes; however, the DNA content of the arrested cells was equivalent to that in G1 phase. After reoxygenation, time-lapse imaging and DNA content analysis revealed that all cells reached G2 phase, and that Fucci fluorescence was distinctly separated into two fractions 24h after reoxygenation: red cells that released from G2 arrest after repairing DNA double-strand breaks (DSBs) exhibited higher clonogenic survival, whereas most cells that stayed green contained many DSBs and exhibited lower survival. We conclude that hypoxia disrupts coordination of DNA synthesis and E3 ligase activities associated with cell-cycle progression, and that DSB repair could greatly influence cell-cycle kinetics and clonogenic survival after hypoxia/reoxygenation.

  18. A New Indicator Cell Line Established to Monitor Bovine Foamy Virus Infection

    Institute of Scientific and Technical Information of China (English)

    Hong-yan Guo; Zhi-bin Liang; Yue Li; Juan Tan; Qi-min Chen; Wen-tao Qiao

    2011-01-01

    In order to improve the accuracy for quantitating the bovine foamy virus(BFV)in vitro,we developed a baby hamster kidney cell(BHK)-21-derived indicator cell line containing a plasmid that encodes the firefly luciferase driven by the BFV long terminal repeat promoter(LTR,from -7 to 1012). The BFV titer could be determined by detecting the luciferase expression since the viral trans-activator BTas protein activates the promoter activity of the LTR. One clone,designated BFVL,was selected from ten neomycin-resistant clones.BFVL showed a specific and inducible dose-and time-dependent luciferase activity in response to BFV infection.Although the changes in luciferase activity of BFVL peaked at 84 h post infection,it was possible to differentiate infected and uninfected cells at 48 h post infection. A linear relationship was established between the multiplicity of infection(MOI)of BFV and the activated ratio of luciferase expression in BFVL. Moreover,the sensitivity of the BFVL-based assay for detecting infectious BFV was 10,000 times higher than the conventional CPE-based assay at 48 h post infection. These findings suggest that the BFVL-based assay is rapid,easy,sensitive,quantitative and specific for detection of BFV infection.

  19. Are Soft Short Tests Good Indicators of Internal Li-ion Cell Defects?

    Science.gov (United States)

    Jeevarajan, J.; Chung, J.-S.; Jung, K.; Park, J.

    2013-01-01

    The self discharge test at full state of charge, may not be a good one to detect subtle defects since the li-ion chemistry has the highest self discharge at full state of charge. One should characterize self discharge versus storage time for each cell manufacturer/design to differentiate between normal self discharge and that due to a subtle manufacturing defect. The various soft short test methods indicate that if this test is carried out at full discharge (0% SOC) with all capacity removed (by lowering the current load in a stepwise manner to the same end of discharge voltage), then the cells need to be placed in storage for more than 72 hours to get a good analysis on the presence of subtle defects since it takes more than 72 hours to achieve voltage stabilization. If the cells are to be charged up even to a small percentage (ex. 1%), 72 hours are sufficient to determine issues. However, the pass/fail criteria should be based on a valid OCV decline. Less than 10 mV voltage decline is not a good method to detect subtle defects. As mentioned in the first bullet, self discharge is a competing reaction when a charge is introduced and hence a characterization of the self discharge versus storage time is required to fully correlate voltage decline to a failure due to a subtle defect. Soft short test method cannot be relied on for defect detection because cells with and without voltage decline seemed to have similar defects and characteristics. Screening methods such as internal resistance and capacity as well as a 3-sigma range for OCV, mass and dimensions should be used to screen out outliers. A very critical aspect in the understanding of subtle defects is to carry out destructive analysis of cells from every lot to confirm the quality of production and screen all cells and batteries in a stringent manner to have a high quality set of flight cells. Self Discharge Test: Fully charged cells shall be placed in Open circuit stand for 72 hours (OCV measurement twice a

  20. Vismodegib in the treatment of basal cell carcinoma: indications for clinical practice.

    Science.gov (United States)

    Calzavara Pinton, Piergiacomo; Licitra, Lisa; Peris, Katia; Santoro, Armando; Ascierto, Paolo Antonio

    2015-01-01

    Basal cell carcinoma (BCC) is a frequent skin cancer which can cause substantial morbidity due to its location on the face, its frequency of relapse and its capacity to invade local tissues. The primary treatment of BCC usually involves surgery or radiotherapy. In patients who have exhausted surgical and radiotherapy options or with metastatic BCC, guidelines recommend the use of the Hedgehog pathway inhibitor vismodegib. This molecule is indicated for the treatment of adults with metastatic BCC, or with locally advanced BCC which has recurred following surgery or who are not eligible to surgery or radiation. This paper aims to provide suggestions on the optimal management of BCC patients treated with vismodegib in clinical practice, according to the large experience gained by a group of Italian dermatologists and oncologists. In particular, the focus of this paper will be on the monitoring of patients and the management of adverse events.

  1. Feces of feedlot cattle contain a diversity of bacteriophages that lyse non-O157 Shiga toxin-producing Escherichia coli.

    Science.gov (United States)

    Wang, Jiaying; Niu, Yan D; Chen, Jinding; Anany, Hany; Ackermann, Hans-W; Johnson, Roger P; Ateba, Collins N; Stanford, Kim; McAllister, Tim A

    2015-07-01

    This study aimed to isolate and characterize bacteriophages that lyse non-O157 Shiga toxin-producing Escherichia coli (STEC) from cattle feces. Of 37 non-O157 STEC-infecting phages isolated, those targeting O26 (AXO26A, AYO26A, AYO26B), O103 (AXO103A, AYO103A), O111 (AXO111A, AYO111A), O121 (AXO121A, AXO121B), and O145 (AYO145A, AYO145B) were further characterized. Transmission electron microscopy showed that the 11 isolates belonged to 3 families and 6 genera: the families Myoviridae (types rV5, T4, ViI, O1), Siphoviridae (type T5), and Podoviridae (type T7). Genome size of the phages as determined by pulsed-field gel electrophoresis ranged from 38 to 177 kb. Excluding phages AXO26A, AYO103A, AYO145A, and AYO145B, all other phages were capable of lysing more than 1 clinically important strain from serogroups of O26, O91, O103, O111, O113, O121, and O128, but none exhibited infectivity across all serogroups. Moreover, phages AYO26A, AXO121A, and AXO121B were also able to lyse 4 common phage types of STEC O157:H7. Our findings show that a diversity of non-O157 STEC-infecting phages are harbored in bovine feces. Phages AYO26A, AYO26B, AXO103A, AXO111A, AYO111A, AXO121A, and AXO121B exhibited a broad host range against a number of serogroups of STEC and have potential for the biocontrol of STEC in the environment.

  2. Photobinding of drugs to cells as an indicator of potential photoallergy.

    Science.gov (United States)

    Miranda, M A; Castell, J V; Gómez-Lechón, M J; Hernández, D; Martínez, L A

    1995-08-01

    The photobinding of drugs to cells has been investigated as a possible indicator of the photoallergic potential. Tiaprofenic acid (TA), carprofen (CP), benoxaprofen (BP) and ibuprofen (IP) were selected as test substances and human fibroblasts as model biological system. Radioactive labelling of the drugs was achieved through alkaline exchange of the alpha-carboxyl hydrogens by tritium, using (3)H(2)O as solvent. When the labelled compounds were co-irradiated with fibroblasts, TA gave rise to the highest amount of radioactivity covalently bound to cells. This also occurred, albeit to a lower extent, with CP and BP; by contrast, no effect was observed for IP. The observed rank order is in agreement with the available in vivo data. The time course of the process was determined for TA, distinguishing between covalent and non-covalent binding. The results, together with gas Chromatographie analyses of the irradiation mixtures and binding studies with TA and its major photoproduct decarboxytiaprofenic acid (decarboxy-TA) in the dark, indicated that most of the observed photobinding might be due to the photoproduct rather than to the parent drug. It is conceivable that hydrogen abstraction by the excited ketone could take place with proteins as reaction partners. This process would lead to the generation of a radical pair, whose coupling would result in the formation of a covalent bond. The contribution of this mechanistic pathway to the in vivo photobinding has to be considered when a lipophilic photoproduct is formed which, as for decarboxy-TA, still contains an active chromophore.

  3. Selection and characterization of a candidate therapeutic bacteriophage that lyses the Escherichia coli O104:H4 strain from the 2011 outbreak in Germany.

    Directory of Open Access Journals (Sweden)

    Maia Merabishvili

    Full Text Available In 2011, a novel strain of O104:H4 Escherichia coli caused a serious outbreak of foodborne hemolytic uremic syndrome and bloody diarrhea in Germany. Antibiotics were of questionable use and 54 deaths occurred. Candidate therapeutic bacteriophages that efficiently lyse the E. coli O104:H4 outbreak strain could be selected rather easily from a phage bank or isolated from the environment. It is argued that phage therapy should be more considered as a potential armament against the growing threat of (resistant bacterial infections.

  4. Laser therapy for upper urinary tract transitional cell carcinoma: indications and management.

    Science.gov (United States)

    Bader, Markus J; Sroka, Ronald; Gratzke, Christian; Seitz, Michael; Weidlich, Patrick; Staehler, Michael; Becker, Armin; Stief, Christian G; Reich, Oliver

    2009-07-01

    Ureteroscopically guided laser techniques are commonly used in the treatment of upper urinary tract transitional cell carcinoma (UUTT); however, there is an ongoing debate with regard to indication and management. To review the indication, feasibility, and treatment outcome of laser application for definitive endoscopic treatment of UUTT, focusing on technical aspects of different laser devices and their impact on tissue. PubMed and Medline were searched for reports on laser therapy in UUTT from 1980 to 2008, with particular focus on the technical background of various laser systems. For decades, nephroureterectomy has been considered the gold standard for treating UUTT. With the intent to preserve functioning renal parenchyma, minimally invasive approaches, initially advocated for patients requiring a nephron-sparing approach (ie, single functioning kidney, renal insufficiency or significant comorbidities), have gained widespread acceptance due to advances in ureteroscopy, percutaneous renal surgery, and laparoscopy. Ureteroscopically guided laser ablation has been used successfully, resulting in recurrence rates ranging from 31% to 65% and disease-free rates of 35% to 86%, depending on stage and grade at diagnosis. To obtain the highest treatment success, the initial staging and grading of the tumour is crucial. Because low-grade tumours rarely if ever progress in stage or grade, the success rate of ureteroscopic therapy parallels that of endoscopic resection of identical bladder tumours. In the treatment of higher grade, advanced tumours, ureteroscopic therapy is less likely to be curative, and thus, endoscopic manoeuvres can only be palliative. Due to the relatively low prevalence of this tumour and the lack of comparable randomised, multicentre trials, the indications for an endoscopic laser treatment option has to be defined based on the patient's individual situation.

  5. Are lung cysts in renal cell cancer (RCC) patients an indication for FLCN mutation analysis?

    Science.gov (United States)

    Johannesma, Paul C; Houweling, Arjan C; Menko, Fred H; van de Beek, I; Reinhard, Rinze; Gille, Johan J P; van Waesberghe, JanHein T M; Thunnissen, Erik; Starink, Theo M; Postmus, Pieter E; van Moorselaar, R Jeroen A

    2016-04-01

    Renal cell cancer (RCC) represents 2-3% of all cancers and is the most lethal of the urologic malignancies, in a minority of cases caused by a genetic predisposition. Birt-Hogg-Dubé syndrome (BHD) is one of the hereditary renal cancer syndromes. As the histological subtype and clinical presentation in BHD are highly variable, this syndrome is easily missed. Lung cysts--mainly under the main carina--are reported to be present in over 90% of all BHD patients and might be an important clue in differentiating between sporadic RCC and BHD associated RCC. We conducted a retrospective study among patients diagnosed with sporadic RCC, wherein we retrospectively scored for the presence of lung cysts on thoracic CT. We performed FLCN mutation analysis in 8 RCC patients with at least one lung cysts under the carina. No mutations were identified. We compared the radiological findings in the FLCN negative patients to those in 4 known BHD patients and found multiple basal lung cysts were present significantly more frequent in FLCN mutation carriers and may be an indication for BHD syndrome in apparent sporadic RCC patients.

  6. Red cell indices for distinguishing macrocytosis of aplastic anaemia and megaloblastic anaemia.

    Science.gov (United States)

    Gupta, P K; Saxena, Renu; Karan, A S; Choudhry, V P

    2003-07-01

    Megaloblastic anaemia and aplastic anaemia are important causes of pancytopenia in India. Since both may have presence of macrocytes, peripheral smear examination alone may pose a difficulty in distinction between the two in the absence of macro-ovalocytes and hypersegmented neutrophils. The present study was conducted to evaluate the role of red cell indices in differentiation between macrocytosis of aplastic anaemia and megaloblastic anaemia. Haemogram from 25 cases each of biopsy proven megaloblastic anaemia and aplastic anaemia were reviewed. It was observed that MCV was greater than 97 fl in 15 cases of aplastic anemia (mean MCV 109.7 fl), and 25 cases of megaloblastic anaemia (mean MCV 113.2 fl). Hb, MCV & MCHC were comparable in the two groups. However, mean RDW in megaloblastic anaemia (mean 87.7 fl) was significantly higher than those in aplastic anaemia (mean 71.4 fl). The difference in RDW of patients with megaloblastic anaemia and aplastic anaemia was statistically significant. We conclude that RDW can be of help to differentiate between the two conditions.

  7. Proliferative index activity in oral squamous cell carcinoma: indication for postoperative radiotherapy?

    Science.gov (United States)

    Gontarz, M; Wyszyńska-Pawelec, G; Zapała, J; Czopek, J; Lazar, A; Tomaszewska, R

    2014-10-01

    The predictive value of the Ki-67 labelling index and its relationship with radiosensitivity in oral squamous cell carcinoma (SCC) remains controversial. We sought to evaluate whether the expression of Ki-67 antigen found in SCC of the tongue and the floor of the mouth is an indication for postoperative radiotherapy (PORT). The first study group included 34 patients who were treated only with primary surgery, while the second group included 26 patients who underwent primary surgery combined with PORT. The correlation between Ki-67 expression and loco-regional recurrence, as well as the 5-year disease-specific survival, was assessed in the two groups. Cases of high-proliferative tumours showed a significantly higher risk of loco-regional recurrence (P=0.018) and a poorer prognosis (P=0.001) only in the 34 patients treated with surgery alone. In multivariate Cox regression analysis, high Ki-67 expression was an independent predictor of loco-regional recurrence (HR 5.42, P=0.029) and disease-specific survival (HR 9.02, P=0.004). The correlation between Ki-67 expression and the risk of loco-regional recurrence in SCC of the tongue and the floor of the mouth may be useful in the selection of patients at a higher risk of recurrence who would benefit from PORT, despite adequate margins of resection and early stage of the disease.

  8. Ratio of spleen diameter to red blood cell distribution width: a novel indicator for celiac disease.

    Science.gov (United States)

    Balaban, Daniel Vasile; Popp, Alina; Lungu, Andrei Marian; Costache, Raluca Simona; Anca, Ioana Alina; Jinga, Mariana

    2015-04-01

    Celiac disease (CD) is currently considerably underdiagnosed, setting the need for developing tools to select patients with probability of CD, who warrant further testing. Red blood cell distribution width (RDW) has been shown in previous studies to be a sensitive predictor for CD, but it lacks specificity. Splenic hypotrophy is also noted frequently in celiac patients. Our aim was to evaluate if spleen diameter to RDW ratio can be used as an indicator for CD. We evaluated 15 newly diagnosed CD patients, 52 patients with inflammatory bowel disease, and 35 patients with irritable bowel syndrome (IBS). We evaluated the differences in spleen diameter, RDW, and their ratio among the four groups. Two-thirds of the CD patients had elevated RDW, compared to 9% in the IBS group. A small spleen was seen in 80% of the celiacs, compared to 21.9% in the ulcerative colitis group, 10% in the Crohn disease group, and 9% in the IBS group. A spleen diameter to RDW ratio under 6 had a sensitivity of 73.3% and specificity of 88.5% in predicting CD, with an AUROC of 0.737. Spleen diameter to RDW ratio is a simple, widely available score, which can be used to select adult patients with probability of CD.

  9. Immunohistochemical Analysis of Scarring Trachoma Indicates Infiltration by Natural Killer and Undefined CD45 Negative Cells.

    Directory of Open Access Journals (Sweden)

    Victor H Hu

    2016-05-01

    Full Text Available The phenotype and function of immune cells infiltrating the conjunctiva in scarring trachoma have yet to be fully characterized. We assessed tissue morphology and immunophenotype of cellular infiltrates found in trachomatous scarring compared to control participants.Clinical assessments and conjunctival biopsy samples were obtained from 34 individuals with trachomatous scarring undergoing trichiasis surgery and 33 control subjects undergoing cataract or retinal detachment surgery. Biopsy samples were fixed in buffered formalin and embedded in paraffin wax. Hematoxylin and eosin (H&E staining was performed for assessment of the inflammatory cell infiltrate. Immunohistochemical staining of single markers on individual sections was performed to identify cells expressing CD3 (T-cells, CD4 (helper T-cells, CD8 (suppressor/cytotoxic T-cells and Natural Killer, NK, cells, NCR1 (NK cells, CD20 (B-cells, CD45 (nucleated hematopoietic cells, CD56 (NK and T-cells, CD68 (macrophages/monocytes and CD83 (mature dendritic cells. The degree of scarring was assessed histologically using cross-polarized light to visualize collagen fibres.Scarring, regardless of clinical inflammation, was associated with increased inflammatory cell infiltrates on H&E and CD45 staining. Scarring was also associated with increased CD8+ and CD56+ cells, but not CD3+ cells, suggestive of a NK cell infiltrate. This was supported by the presence of NCR1+ cells. There was some increase in CD20+ cells, but no evidence for increased CD4+, CD68+ or CD83+ cells. Numerous CD45 negative cells were also seen in the population of infiltrating inflammatory cells in scarred conjunctiva. Disorganization of the normal collagen architecture was strongly associated with clinical scarring.These data point to the infiltration of immune cells with a phenotype suggestive of NK cells in conjunctival trachomatous scarring. A large proportion of CD45 negative inflammatory cells were also present. Future work

  10. Employment Status as an Indicator of Recovery and Function One Year after Hematopoietic Stem Cell Transplantation.

    Science.gov (United States)

    Morrison, Eleshia J; Ehlers, Shawna L; Bronars, Carrie A; Patten, Christi A; Brockman, Tabetha A; Cerhan, James R; Hogan, William J; Hashmi, Shahrukh K; Gastineau, Dennis A

    2016-09-01

    Employment after hematopoietic stem cell transplantation (HSCT) is an indicator of post-transplantation recovery and function, with economic and social implications. As survival rates for HSCT continue to improve, greater emphasis can be placed on factors affecting the quality of post-transplantation survival, including the ability to resume employment. A sample of recipients of autologous or allogeneic HSCT was accrued (n = 1000) to complete a longitudinal lifestyle survey before transplantation and at 1 year after transplantation. The present study examines associations between employment and patient characteristics, disease variables, illness status, and quality of life among 1-year survivors (n = 702). Participants had a mean age of 55 years (range, 18 to 78) and were predominately male (59.7%), married/partnered (77.1%), and non-Hispanic Caucasian (89.5%); most (79.4%) had received autologous transplantation. Of the 690 participants reporting some form of employment before illness diagnosis, 62.4% had returned to work by 1 year after HSCT. Full-time employment at 1 year after HSCT was significantly associated with remission of illness, improved illness, fewer post-transplantation hospitalizations, less fatigue and pain, higher quality of life, and higher rating of perceived health. Those unemployed because of their health reported the highest rates of fatigue and pain and lowest quality of life, and they were most likely to report poor perceived health. These findings highlight work reintegration as an important outcome and marker of survivors' overall adjustment after transplantation. Identifying factors affecting post-transplantation employment offers opportunities for behavioral interventions to target modifiable risk factors to optimize post-transplantation survivorship, inclusive of increased rates of return to work and decreased rates of associated disability.

  11. Protein ligand-tethered synthetic calcium indicator for localization control and spatiotemporal calcium imaging in plant cells.

    Science.gov (United States)

    Takaoka, Yousuke; Shigenaga, Miyuki; Imai, Masaki; Nukadzuka, Yuuki; Ishimaru, Yasuhiro; Saito, Kei; Yokoyama, Ryusuke; Nishitani, Kazuhiko; Ueda, Minoru

    2016-01-01

    In plant biology, calcium ions are involved in a variety of intriguing biological phenomena as a secondary messenger. However, most conventional calcium indicators are not applicable for plant cells because of the difficulty with their localization control in plant cells. We here introduce a method to monitor spatiotemporal Ca(2+) dynamics in living plant cells based on linking the synthetic calcium indicator Calcium Green-1 to a natural product-based protein ligand. In a proof-of-concept study using cultured BY-2 cells overexpressing the target protein for the ligand, the ligand-tethered probe accumulated in the cytosol and nucleus, and enabled real-time monitoring of the cytosolic and nucleus Ca(2+) dynamics under the physiological condition. The present strategy using ligand-tethered fluorescent sensors may be successfully applied to reveal the spatiotemporal dynamics of calcium ions in living plant cells.

  12. Host natural killer immunity is a key indicator of permissiveness for donor cell engraftment in patients with severe combined immunodeficiency☆

    Science.gov (United States)

    Hassan, Amel; Lee, Pamela; Maggina, Paraskevi; Xu, Jin Hua; Moreira, Diana; Slatter, Mary; Nademi, Zohreh; Worth, Austen; Adams, Stuart; Jones, Alison; Cale, Catherine; Allwood, Zoe; Rao, Kanchan; Chiesa, Robert; Amrolia, Persis; Gaspar, Hubert; Davies, E. Graham; Veys, Paul; Gennery, Andrew; Qasim, Waseem

    2014-01-01

    Background Severe combined immunodeficiency (SCID) can be cured by using allogeneic hematopoietic stem cell transplantation, and the absence of host immunity often obviates the need for preconditioning. Depending on the underlying genetic defect and when blocks in differentiation occur during lymphocyte ontogeny, infants with SCID have absent or greatly reduced numbers of functional T cells. Natural killer (NK) cell populations are usually absent in the SCID-X1 and Janus kinase 3 forms of SCID and greatly reduced in adenosine deaminase deficiency SCID but often present in other forms of the disorder. Objective To determine if SCID phenotypes indicate host permissiveness to donor cell engraftment. Methods A retrospective data analysis considered whether host NK cells influenced donor T-cell engraftment, immune reconstitution, and long-term outcomes in children who had undergone nonconditioned allogeneic stem cell transplantation between 1990 and 2011 in the United Kingdom. Detailed analysis of T- and B-cell immune reconstitution and donor chimerism was compared between the NK+ (n = 24) and NK− (n = 53) forms of SCID. Results Overall, 77 children underwent transplantation, with survival of 90% in matched sibling donor/matched family donor transplants compared with 60% when alternative donors were used. Infants with NK−SCID were more likely to survive than NK+ recipients (87% vs 62%, P < .01) and had high-level donor T-cell chimerism with superior long-term recovery of CD4 T-cell immunity. Notably, 33% of children with NK+SCID required additional transplantation procedures compared with only 8% of children with NK−SCID (P < .005). Conclusions NK−SCID disorders are highly permissive for donor T-cell engraftment without preconditioning, whereas the presence of NK cells is a strong indicator that preparative conditioning is required for engraftment of T-cell precursors capable of supporting robust T-cell reconstitution. PMID:24794685

  13. Expression Patterns of Cancer-Testis Antigens in Human Embryonic Stem Cells and Their Cell Derivatives Indicate Lineage Tracks

    OpenAIRE

    Nadya Lifantseva; Anna Koltsova; Tatyana Krylova; Tatyana Yakovleva; Galina Poljanskaya; Olga Gordeeva

    2011-01-01

    Pluripotent stem cells can differentiate into various lineages but undergo genetic and epigenetic changes during long-term cultivation and, therefore, require regular monitoring. The expression patterns of cancer-testis antigens (CTAs) MAGE-A2, -A3, -A4, -A6, -A8, -B2, and GAGE were examined in undifferentiated human embryonic stem (hES) cells, their differentiated derivatives, teratocarcinoma (hEC) cells, and cancer cell lines of neuroectodermal and mesodermal origin. Undifferentiated hES ce...

  14. ARMc8 indicates aggressive colon cancers and promotes invasiveness and migration of colon cancer cells.

    Science.gov (United States)

    Jiang, Guiyang; Zhang, Yong; Zhang, Xiupeng; Fan, Chuifeng; Wang, Liang; Xu, Hongtao; Yu, Juanhan; Wang, Enhua

    2015-11-01

    Recent studies have implicated ARMc8 in promoting tumor formation in non-small cell lung cancer and breast cancer; however, so far, no studies have revealed the expression pattern or cellular function of ARMc8 in colon cancer. In this study, we used immunohistochemical staining to measure ARMc8 expression in 206 cases of colon cancer and matched adjacent normal colon tissue. Clinically important behaviors of cells, including invasiveness and migration, were evaluated after upregulation of ARMc8 expression in HT29 cells through gene transfection or downregulation of expression in LoVo cells using RNAi. We found that ARMc8 was primarily located in the membrane and cytoplasm of tumor cells, and its expression level was significantly higher in colon cancer in comparison to that in the adjacent normal colon tissues (p TNM stage (p = 0.006), lymph node metastasis (p = 0.001), and poor prognosis (p = 0.002) of colon cancer. The invasiveness and migration capacity of HT29 cells transfected with ARMc8 were significantly greater than those of control cells (p colon cancer cells. ARMc8 is likely to become a potential therapeutic target for colon cancer.

  15. Indicator cell lines for the detection of hidden mycoplasma contamination, using an adenosine phosphorylase screening test

    NARCIS (Netherlands)

    Spierenburg, G.T.; Polak-Vogelzang, A.A.; Bast, B.J.E. G.

    Mycoplasmas are a major cause of cell culture contamination and are especially troublesome during HAT selection. The enzyme adenosine phosphorylase (adoP) is present in all common mycoplasma species but is considered to have a low activity in mammalian cells. However, using an adoP screening test,

  16. Mean-cluster approach indicates cell sorting time scales are determined by collective dynamics

    Science.gov (United States)

    Beatrici, Carine P.; de Almeida, Rita M. C.; Brunnet, Leonardo G.

    2017-03-01

    Cell migration is essential to cell segregation, playing a central role in tissue formation, wound healing, and tumor evolution. Considering random mixtures of two cell types, it is still not clear which cell characteristics define clustering time scales. The mass of diffusing clusters merging with one another is expected to grow as td /d +2 when the diffusion constant scales with the inverse of the cluster mass. Cell segregation experiments deviate from that behavior. Explanations for that could arise from specific microscopic mechanisms or from collective effects, typical of active matter. Here we consider a power law connecting diffusion constant and cluster mass to propose an analytic approach to model cell segregation where we explicitly take into account finite-size corrections. The results are compared with active matter model simulations and experiments available in the literature. To investigate the role played by different mechanisms we considered different hypotheses describing cell-cell interaction: differential adhesion hypothesis and different velocities hypothesis. We find that the simulations yield normal diffusion for long time intervals. Analytic and simulation results show that (i) cluster evolution clearly tends to a scaling regime, disrupted only at finite-size limits; (ii) cluster diffusion is greatly enhanced by cell collective behavior, such that for high enough tendency to follow the neighbors, cluster diffusion may become independent of cluster size; (iii) the scaling exponent for cluster growth depends only on the mass-diffusion relation, not on the detailed local segregation mechanism. These results apply for active matter systems in general and, in particular, the mechanisms found underlying the increase in cell sorting speed certainly have deep implications in biological evolution as a selection mechanism.

  17. Effect of pH on biologic degradation of Microcystis aeruginosa by alga-lysing bacteria in sequencing batch biofilm reactors

    Institute of Scientific and Technical Information of China (English)

    Hongjing LI; Mengli HAO; Jingxian LIU; Chen CHEN1; Zhengqiu FAN; Xiangrong WANG

    2012-01-01

    In this paper, the effect of pH on biological degradation of Microcystis aeruginosa by alga-lysing bacteria in laboratory-scale sequencing batch biofilm reactors (SBBRs) was investigated. After 10 d filming with waste activated sludge, the biological film could be formed, and the bioreactors in which laid polyolefin resin filler were used to treat algal culture. By comparing the removal efficiency of chlorophyll a at different aerobic time, the optimum time was determined as 5 h. Under pH 6.5, 7.5, and 8.5 conditions, the removal rates of Microcystis aeruginosa were respectively 75.9%, 83.6%, and 78.3% (in term of chlorophyll a), and that of Chemical Oxygen Demand (CODMn) were 30.6%, 35.8%, and 33.5%. While the removal efficiencies of ammonia nitrogen (NH+ -N) were all 100%. It was observed that the sequence of the removal efficiencies of algae, NH+ -N and organic matter were pH 7.5 〉 pH 8.5 〉 pH 6.5. The results showed that the dominant alga-lysing bacteria in the SBBRs was strain HM-01, which was identified as Bacillus sp. by Polymerase Chain Reaction (PCR) amplification of the 16S rRNA gene, Basic Local Alignment Search Tool (BLAST) analysis, and compar- ison with sequences in the GenBank nucleotide database. The algicidal activated substance which HM-01 strain excreted could withstand high temperature and pressure, also had better hydrophily and stronger polarity.

  18. Expression of Immunoglobulin Receptors with Distinctive Features Indicating Antigen Selection by Marginal Zone B Cells from Human Spleen

    Science.gov (United States)

    Colombo, Monica; Cutrona, Giovanna; Reverberi, Daniele; Bruno, Silvia; Ghiotto, Fabio; Tenca, Claudya; Stamatopoulos, Kostas; Hadzidimitriou, Anastasia; Ceccarelli, Jenny; Salvi, Sandra; Boccardo, Simona; Calevo, Maria Grazia; De Santanna, Amleto; Truini, Mauro; Fais, Franco; Ferrarini, Manlio

    2013-01-01

    Marginal zone (MZ) B cells, identified as surface (s)IgMhighsIgDlowCD23low/−CD21+CD38− B cells, were purified from human spleens, and the features of their V(D)J gene rearrangements were investigated and compared with those of germinal center (GC), follicular mantle (FM) and switched memory (SM) B cells. Most MZ B cells were CD27+ and exhibited somatic hypermutations (SHM), although to a lower extent than SM B cells. Moreover, among MZ B-cell rearrangements, recurrent sequences were observed, some of which displayed intraclonal diversification. The same diversifying sequences were detected in very low numbers in GC and FM B cells and only when a highly sensitive, gene-specific polymerase chain reaction was used. This result indicates that MZ B cells could expand and diversify in situ and also suggested the presence of a number of activation-induced cytidine deaminase (AID)-expressing B cells in the MZ. The notion of antigen-driven expansion/selection in situ is further supported by the VH CDR3 features of MZ B cells with highly conserved amino acids at specific positions and by the finding of shared (“stereotyped”) sequences in two different spleens. Collectively, the data are consistent with the notion that MZ B cells are a special subset selected by in situ antigenic stimuli. PMID:23877718

  19. Photoacoustic detection of hemozoin in human mononuclear cells as an early indicator of malaria infection

    Science.gov (United States)

    Custer, Jonathan R.; Kariuki, Michael; Beerntsen, Brenda T.; Viator, John A.

    2010-02-01

    Malaria is a blood borne infection affecting hundreds of millions of people worldwide2. The parasites reproduce within the blood cells, eventually causing their death and lysis. This process releases the parasites into the blood, continuing the cycle of infection. Usually, malaria is diagnosed only after a patient presents symptoms, including high fever, nausea, and, in advanced cases, coma and death. While invading the bloodstream of a host, malaria parasites convert hemoglobin into an insoluble crystal, known as hemozoin. These crystals, approximately several hundred nanometers in size, are contained within red blood cells and white blood cells that ingest free hemozoin in the blood. Thus, infected red blood cells and white blood cells contain a unique optical absorber that can be detected in blood samples using static photoacoustic detection methods. We separated the white blood cells from malaria infected blood and tested it in a photoacoustic set up using a tunable laser system consisting of an optical parametric oscillator pumped by an Nd:YAG laser with pulse duration of 5 ns. Our threshold of detection was 10 infected white blood cells per microliter, which is more sensitive than current diagnosis methods using microscopic analysis of blood.

  20. Inflammatory mediators and cell adhesion molecules as indicators of severity of atherosclerosis: the Rotterdam Study

    NARCIS (Netherlands)

    M.P.M. de Maat (Moniek); M.L. Bots (Michiel); M.M.B. Breteler (Monique); J. Meijer (John); A.J. Kiliaan (Amanda); J.C.M. Witteman (Jacqueline); A. Hofman (Albert)

    2002-01-01

    textabstractInflammatory mediators and soluble cell adhesion molecules predict cardiovascular events. It is not clear whether they reflect the severity of underlying atherosclerotic disease. Within the Rotterdam Study, we investigated the associations of C-reactive protein (CRP), i

  1. Parametric Response Mapping as an Indicator of Bronchiolitis Obliterans Syndrome after Hematopoietic Stem Cell Transplantation

    NARCIS (Netherlands)

    Galban, Craig J.; Boes, Jennifer L.; Bule, Maria; Kitko, Carrie L.; Couriel, Daniel R.; Johnson, Timothy D.; Lama, Vihba; Telenga, Eef D.; van den Berge, Maarten; Rehemtulla, Alnawaz; Kazerooni, Ella A.; Ponkowski, Michael J.; Ross, Brian D.; Yanik, Gregory A.

    2014-01-01

    The management of bronchiolitis obliterans syndrome (BOS) after hematopoietic cell transplantation presents many challenges, both diagnostically and therapeutically. We developed a computed tomography (CT) voxel-wise methodology termed parametric response mapping (PRM) that quantifies normal

  2. Can Lucifer Yellow Indicate Correct Permeability of Biological Cell Membrane under An Electric and Magnetic Field?

    Directory of Open Access Journals (Sweden)

    Tahereh Pourmirjafari Firoozabadi

    2015-01-01

    Full Text Available The effect of external magnetic and electric fields, in the range of electroporation and magnetoporation, on Lucifer Yellow (LY fluorescence in the absence of cells is studied. Electric-field-induced quenching and magnetic field-induced increase are observed for fluorescence intensity of LY. Regard to the fact that the variation of field-induced fluorescence, even in the absence of cells, can be observed, the application of LY, as a marker, is debatable in electroporation and magnetoporation techniques.

  3. Daily genetic profiling indicates JAK/STAT signaling promotes early hepatic stellate cell transdifferentiation

    Institute of Scientific and Technical Information of China (English)

    Ashley; M; Lakner; Cathy; C; Moore; Alyssa; A; Gulledge; Laura; W; Schrum

    2010-01-01

    AIM: To identify signaling pathways and genes that initiate and commit hepatic stellate cells (HSCs) to transdifferentiation. METHODS: Primary HSCs were isolated from male Sprague-Dawley rats and cultured on plastic for 0-10 d. Gene expression was assessed daily (quiescent to day 10 culture-activation) by real time polymerase chain reaction and data clustered using AMADA software. The significance of JAK/STAT signaling to HSC transdifferentiation was determined by treating cells with a JAK2 inhibitor. RESUL...

  4. Multiple Applications of Alamar Blue as an Indicator of Metabolic Function and Cellular Health in Cell Viability Bioassays

    Directory of Open Access Journals (Sweden)

    Sephra N. Rampersad

    2012-09-01

    Full Text Available Accurate prediction of the adverse effects of test compounds on living systems, detection of toxic thresholds, and expansion of experimental data sets to include multiple toxicity end-point analysis are required for any robust screening regime. Alamar Blue is an important redox indicator that is used to evaluate metabolic function and cellular health. The Alamar Blue bioassay has been utilized over the past 50 years to assess cell viability and cytotoxicity in a range of biological and environmental systems and in a number of cell types including bacteria, yeast, fungi, protozoa and cultured mammalian and piscine cells. It offers several advantages over other metabolic indicators and other cytotoxicity assays. However, as with any bioassay, suitability must be determined for each application and cell model. This review seeks to highlight many of the important considerations involved in assay use and design in addition to the potential pitfalls.

  5. Immunolocalization indicates that both original and regenerated lizard tail tissues contain populations of long retaining cells, putative stem/progenitor cells.

    Science.gov (United States)

    Alibardi, Lorenzo

    2015-11-01

    The regeneration of the tail in lizards is likely sustained by stem/progenitor cells located in the stump after amputation of the tail. This microscopic and ultrastructural study shows the localization of 5-bromo-deoxy-uridine (5BrdU)-long retaining labeled cells in different tissues of the tail stump. These putative stem/progenitor cells are sparsely detected in the epidermis of scales, adipose tissue, intermuscle connective septa, myosatellite cells, and perichondrion of the vertebrae. Most of 5BrdU-labeled cells are present in the bone marrow of vertebrae as hemocytoblasts and reticulate cells, whereas more numerous myelocytes and polychromatophilic erythroblasts show a variable level of nuclear labeling. 5BrdU and tritiated-thymidine labeled and unlabeled hemopoietic cells are seen in circulating vessels and in the blastema where their maturation is completed. This observation indicates that the entire differentiation span of both white and red blood cells, at least during tail regeneration, lasts longer than 4 weeks. Labeled polychromatophilic erythroblasts and heterophilic and basophilic myelocytes are present in the synusoidal vessels of the regenerating tail. This study indicates that extravasating blood cells involved in immunity make large part of the forming blastema cell population, but are replaced by mesenchymal cells of different origin. The presence of long retaining labeled cells in tissues of the tail stump is likely connected to the production of blastema mesenchymal cells. Although no direct cell-lineage study has been done, histological, immunocytochemical, and autoradiographic studies have indicated that it is from these tissues that proliferating cells appear mainly localized after tail amputation and blastema formation.

  6. Swelling of root cell walls as an indicator of their functional state.

    Science.gov (United States)

    Meychik, N R; Yermakov, I P

    2001-02-01

    The swelling capacity of cell walls isolated from different parts of lupine root was investigated. The water content in fragments of intact roots (Q) and swelling coefficient of standardized samples of cell walls (Kcw) were determined, and the dependences of Q and Kcw on the distance from the root tip (L) were plotted. It was shown that the change in Q value along the stretch of the lupine root reaches its maximum at distances of 1.5-6 cm or 7-12 cm from the root tip in 7-day-old and 14-day-old seedlings, respectively, whereas the Kcw value distribution over the root length is virtually invariable. In the radial direction, both the Q and Kcw values in cortex tissues are about twice higher than in the central cylinder. In our opinion, the changes of both Q and Kcw in the radial direction are associated with different degrees of cross-linking between polymer chains in cell wall structures of root cortex and central cylinder. The results of measurement of the Kcw value are consistent with the widely accepted mechanisms of water transport in roots in the radial direction. These data show that water transport through apoplast to the border between the cortex and central cylinder is accompanied by an increase in the resistance to water flow. Among other factors, this increase is due to a greater degree of cross-linking between cell wall polymers in the central cylinder. The results of measurement of the swelling coefficient of standardized cell wall samples in water and in 10 mM KCl at different pH values show that the swelling capacity of root cell walls varies according to the physicochemical properties of synthetic ion exchangers. Cell walls shrink (cell wall volume decreases) as ion concentration in solution increases and pH decreases. This causes an increase in the hydraulic resistance (or a decrease in the hydraulic conductivity) of apoplast. It was concluded that swelling is determined by the physicochemical properties of the cell wall, whereas the change in the

  7. Inhibition of cell adhesion by xARVCF indicates a regulatory function at the plasma membrane.

    Science.gov (United States)

    Reintsch, Wolfgang E; Mandato, Craig A; McCrea, Pierre D; Fagotto, François

    2008-09-01

    The cytoplasmic tail of cadherins is thought to regulate the strength and dynamics of cell-cell adhesion. Part of its regulatory activity has been attributed to a membrane-proximal region, the juxtamembrane domain (JMD), and its interaction with members of the p120 catenin subfamily. We show that titration of xARVCF, a member of this family, to the plasma membrane disrupts adhesion in the early embryo. Adhesion can be restored by coexpression of constitutively active Rac, suggesting that intracellular signaling is the primary cause in the loss of adhesion phenotype. Our observations suggest that the recruitment of p120 type catenins to the plasma membrane by the cadherin cytoplasmic tail may create protein complexes, which actively modulate the adhesion "status" of embryonic cells.

  8. Global epigenomic analysis indicates protocadherin-7 activates osteoclastogenesis by promoting cell–cell fusion

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, Haruhiko [Department of Orthopaedic Surgery, Faculty of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033 (Japan); Department of Cell Signaling, Graduate School of Medical and Dental Science, Tokyo Medical and Dental University, Yushima 1-5-45, Bunkyo-ku, Tokyo 113-8549 (Japan); Nakashima, Tomoki [Department of Cell Signaling, Graduate School of Medical and Dental Science, Tokyo Medical and Dental University, Yushima 1-5-45, Bunkyo-ku, Tokyo 113-8549 (Japan); Japan Science and Technology Agency, PRESTO, Yushima 1-5-45, Bunkyo-ku, Tokyo 113-8549 (Japan); Hayashi, Mikihito [Department of Cell Signaling, Graduate School of Medical and Dental Science, Tokyo Medical and Dental University, Yushima 1-5-45, Bunkyo-ku, Tokyo 113-8549 (Japan); Japan Science and Technology Agency, ERATO, Takayanagi Osteonetwork Project, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Izawa, Naohiro; Yasui, Tetsuro [Department of Orthopaedic Surgery, Faculty of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033 (Japan); Aburatani, Hiroyuki [Genome Science Division, Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1, Komaba, Meguro-ku, Tokyo 153-8904 (Japan); Tanaka, Sakae [Department of Orthopaedic Surgery, Faculty of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033 (Japan); Takayanagi, Hiroshi, E-mail: takayana@m.u-tokyo.ac.jp [Japan Science and Technology Agency, ERATO, Takayanagi Osteonetwork Project, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Department of Immunology, Graduate School of Medicine and Faculty of Medicine, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)

    2014-12-12

    Highlights: • Identification of epigenetically regulated genes during osteoclastogenesis. • Pcdh7 is regulated by H3K4me3 and H3K27me3 during osteoclastogenesis. • Pcdh7 expression is increased by RANKL during osteoclastogenesis. • Establishment of novel cell fusion analysis for osteoclasts by imaging cytometer. • Pcdh7 regulates osteoclastogenesis by promoting cell fusion related gene expressions. - Abstract: Gene expression is dependent not only on genomic sequences, but also epigenetic control, in which the regulation of chromatin by histone modification plays a crucial role. Histone H3 lysine 4 trimethylation (H3K4me3) and histone H3 lysine 27 trimethylation (H3K27me3) are related to transcriptionally activated and silenced sequences, respectively. Osteoclasts, the multinucleated cells that resorb bone, are generated by the fusion of precursor cells of monocyte/macrophage lineage. To elucidate the molecular and epigenetic regulation of osteoclast differentiation, we performed a chromatin immunoprecipitation sequencing (ChIP-seq) analysis for H3K4me3 and H3K27me3 in combination with RNA sequencing. We focused on the histone modification change from H3K4me3(+)H3K27me3(+) to H3K4me3(+)H3K27me3(–) and identified the protocadherin-7 gene (Pcdh7) to be among the genes epigenetically regulated during osteoclastogenesis. Pcdh7 was induced by RANKL stimulation in an NFAT-dependent manner. The knockdown of Pcdh7 inhibited RANKL-induced osteoclast differentiation due to the impairment of cell–cell fusion, accompanied by a decreased expression of the fusion-related genes Dcstamp, Ocstamp and Atp6v0d2. This study demonstrates that Pcdh7 plays a key role in osteoclastogenesis by promoting cell–cell fusion.

  9. Differences in endolymphatic sac mitochondria-rich cells indicate specific functions.

    NARCIS (Netherlands)

    Peters, T.A.; Tonnaer, E.L.G.M.; Kuijpers, W.; Cremers, C.W.R.J.; Curfs, J.H.A.J.

    2002-01-01

    OBJECTIVE/HYPOTHESIS: The purpose of the study was to examine the specific involvement of endolymphatic sac mitochondria-rich cells in endolymph homeostasis. STUDY DESIGN: Transmission electron microscopy and immunohistochemistry were performed on the endolymphatic sac of young adult rats, and two

  10. Lack of indication of myocardial cell damage after myocardial ischaemia in patients with severe stable angina

    DEFF Research Database (Denmark)

    Hansen, Knud Nørregaard; Egstrup, K; Nielsen, J R

    1992-01-01

    To evaluate myocardial cell damage in relation to spontaneous and exercise-induced ischaemia, release of myoglobin, creatine kinase (CK) and its isoenzyme MB (CK-MB) into the serum was estimated in 10 patients with severe stable angina. All patients had a positive exercise test, significant steno...

  11. Stem cell differentiation indicated by noninvasive photonic characterization and fractal analysis of subcellular architecture.

    Science.gov (United States)

    Chalut, Kevin J; Kulangara, Karina; Wax, Adam; Leong, Kam W

    2011-08-01

    We hypothesised that global structural changes in stem cells would manifest with differentiation, and that these changes would be observable with light scattering microscopy. Analysed with a fractal dimension formalism, we observed significant structural changes in differentiating human mesenchymal stem cells within one day after induction, earlier than could be detected by gene expression profiling. Moreover, light scattering microscopy is entirely non-perturbative, so the same sample could be monitored throughout the differentiation process. We explored one possible mechanism, chromatin remodelling, to account for the changes we observed. Correlating with the staining of HP1α, a heterochromatin protein, we applied novel microscopy methods and fractal analysis to monitor the plastic dynamics of chromatin within stem cell nuclei. We showed that the level of chromatin condensation changed during differentiation, and provide one possible explanation for the changes seen with the light scattering method. These results lend physical insight into stem cell differentiation while providing physics-based methods for non-invasive detection of the differentiation process.

  12. Association of ABO Incompatibility With Red Blood Cell Indices of Cord Blood Unit

    Directory of Open Access Journals (Sweden)

    Shu-Huey Chen

    2012-04-01

    Conclusion: Group A or B neonates with a group O mother have lower mean Hb, Hct, and RBC values for CBUs than other neonates. The role of RBC indices in predicting neonatal hemolytic hyperbilirubinemia remains unclear and further studies are needed to identify the possible clinical association.

  13. Imaging the nanomolar range of nitric oxide with an amplifier-coupled fluorescent indicator in living cells

    Science.gov (United States)

    Sato, Moritoshi; Hida, Naoki; Umezawa, Yoshio

    2005-10-01

    Nitric oxide (NO) is a small uncharged free radical that is involved in diverse physiological and pathophysiological mechanisms. NO is generated by three isoforms of NO synthase, endothelial, neuronal, and inducible ones. When generated in vascular endothelial cells, NO plays a key role in vascular tone regulation, in particular. Here, we describe an amplifier-coupled fluorescent indicator for NO to visualize physiological nanomolar dynamics of NO in living cells (detection limit of 0.1 nM). This genetically encoded high-sensitive indicator revealed that 1 nM of NO, which is enough to relax blood vessels, is generated in vascular endothelial cells even in the absence of shear stress. The nanomolar range of basal endothelial NO thus revealed appears to be fundamental to vascular homeostasis. fluorescence resonance energy transfer | genetic encoding

  14. Thalidomide induced early gene expression perturbations indicative of human embryopathy in mouse embryonic stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Gao, Xiugong, E-mail: xiugong.gao@fda.hhs.gov; Sprando, Robert L.; Yourick, Jeffrey J.

    2015-08-15

    Developmental toxicity testing has traditionally relied on animal models which are costly, time consuming, and require the sacrifice of large numbers of animals. In addition, there are significant disparities between human beings and animals in their responses to chemicals. Thalidomide is a species-specific developmental toxicant that causes severe limb malformations in humans but not in mice. Here, we used microarrays to study transcriptomic changes induced by thalidomide in an in vitro model based on differentiation of mouse embryonic stem cells (mESCs). C57BL/6 mESCs were allowed to differentiate spontaneously and RNA was collected at 24, 48, and 72 h after exposure to 0.25 mM thalidomide. Global gene expression analysis using microarrays revealed hundreds of differentially expressed genes upon thalidomide exposure that were enriched in gene ontology (GO) terms and canonical pathways associated with embryonic development and differentiation. In addition, many genes were found to be involved in small GTPases-mediated signal transduction, heart development, and inflammatory responses, which coincide with clinical evidences and may represent critical embryotoxicities of thalidomide. These results demonstrate that transcriptomics in combination with mouse embryonic stem cell differentiation is a promising alternative model for developmental toxicity assessment. - Highlights: • Studied genomic changes in mouse embryonic stem cells upon thalidomide exposure • Identified gene expression changes that may represent thalidomide embryotoxicity • The toxicogenomic changes coincide well with known thalidomide clinical outcomes. • The mouse embryonic stem cell model is suitable for developmental toxicity testing. • The model has the potential for high-throughput screening of a multitude of compounds.

  15. Nuclear expression of β-catenin and stem cell markers as potential prognostic indicators in medulloblastoma

    Directory of Open Access Journals (Sweden)

    Kiran Krishne Gowda

    2012-01-01

    Full Text Available Aims: To study the prognostic role of β-catenin and stem cell markers in medulloblastoma (MB. Materials and Methods: Sixty cases of MB were retrospectively analyzed to study the expression of β-catenin, CD15, and CD133 by immunohistochemistry. Their expression was correlated with histological subtypes and event-free survival (EFS. Patients were divided into Group 1 and 2 based on non-occurrence and occurrence of events during the follow-up period. Results: Fifty of the 60 cases were of classic type of MB while nine were of desmoplastic subtype and one case showed chondroid and rhabdomyoblastic differentiation. Immunoreactivity for β-catenin was observed as nuclear and/or cytoplasmic positivity within the tumor cells. Forty-one (68.3% cases showed cytoplasmic positivity, while nuclear positivity was seen in 21 (35% cases. There was a significant correlation between nuclear expression of β-catenin and different histological subtypes by Chi-square test (P value<0.05. A statistically significant positive correlation of β-catenin nuclear positivity with EFS was observed. Among 60 cases, 37 cases (67.3% showed presence of CD15+ tumor cells with percentage of positivity varying between 0.1 to 17.1%. Overall, 42 of 60 (70% cases showed presence of CD133+ cells. The percentage of positivity varied between 0.1 to 16.5%. A statistically significant negative correlation of CD15 and CD133 positivity with EFS was observed. Conclusions: Nucleopositive β-catenin cases were associated with a favorable outcome on univariate analysis. Both CD15 and CD133 positivity were associated with a worse outcome on univariate analysis.

  16. Expression Analysis of the Hippo Cascade Indicates a Role in Pituitary Stem Cell Development

    Directory of Open Access Journals (Sweden)

    Emily J Lodge

    2016-03-01

    Full Text Available The pituitary gland is a primary endocrine organ that controls major physiological processes. Abnormal development or homeostatic disruptions can lead to human disorders such as hypopituitarism or tumours. Multiple signalling pathways, including WNT, BMP, FGF and SHH regulate pituitary development but the role of the Hippo-YAP1/TAZ cascade is currently unknown. In multiple tissues, the Hippo kinase cascade underlies neoplasias; it influences organ size through the regulation of proliferation and apoptosis, and has roles in determining stem cell potential. We have used a sensitive mRNA in situ hybridisation method (RNAscope to determine the expression patterns of the Hippo pathway components during mouse pituitary development. We have also carried out immunolocalisation studies to determine when YAP1 and TAZ, the transcriptional effectors of the Hippo pathway, are active. We find that YAP1/TAZ are active in the stem/progenitor cell population throughout development and at postnatal stages, consistent with their role in promoting the stem cell state. Our results demonstrate for the first time the collective expression of major components of the Hippo pathway during normal embryonic and postnatal development of the pituitary gland.

  17. Indications on suitable scaffold as carrier of stem cells in the alveoloplasty of cleft palate.

    Science.gov (United States)

    Paganelli, C; Fontana, P; Porta, F; Majorana, A; Pazzaglia, U E; Sapelli, P L

    2006-08-01

    Autologous iliac crest bone is used to close the residual alveolar bone defect in cleft palate patients during late mixed dentition. Surgery involves physical and anaesthesiologic risks, long-time hospitalization, high costs and not always good results (15% failure rate). Alternatives to iliac crest bone grafting are going to be evaluated: synthetic, xenograft and allograft matrices combined with platelet-rich plasma or recombined bone morphogenic proteins for osteoinductivity are commercially available. These alternatives have not yet been determined to be equivalent to the previous treatment. A new field of research is represented by stem cells, which have been also used to regenerate ischaemic cardiac tissue after heart attack, to treat hypophosphatasia and osteoporosis. Our aim was to use osteoblasts from stem cells to close the residual palate cleft in association with a suitable carrier. Stem cells are expanded in the Aastrom bioreactor, differentiated into osteoblasts and positioned in the bone defect by means of a Spongostan scaffold. This scaffold has the best characteristics as commercial availability, low cost, good manageability, absence of allergic reactions or other side effects on patient, biocompatibility, imbibition, radiotransparency, reabsorbability and osteoinductivity. Previous studies encourage Spongostan scaffold application.

  18. Inverse associations between obesity indicators and thymic T-cell production levels in aging atomic-bomb survivors.

    Science.gov (United States)

    Yoshida, Kengo; Nakashima, Eiji; Kubo, Yoshiko; Yamaoka, Mika; Kajimura, Junko; Kyoizumi, Seishi; Hayashi, Tomonori; Ohishi, Waka; Kusunoki, Yoichiro

    2014-01-01

    Reduction of the naive T-cell population represents a deteriorating state in the immune system that occurs with advancing age. In animal model studies, obesity compromises the T-cell immune system as a result of enhanced adipogenesis in primary lymphoid organs and systemic inflammation. In this study, to test the hypothesis that obesity may contribute to the aging of human T-cell immunity, a thousand atomic-bomb survivors were examined for obesity status and ability to produce naive T cells, i.e., T-cell receptor excision circle (TREC) numbers in CD4 and CD8 T cells. The number of TRECs showed a strong positive correlation with naive T cell numbers, and lower TREC numbers were associated with higher age. We found that the TREC number was inversely associated with levels of obesity indicators (BMI, hemoglobin A1c) and serum CRP levels. Development of type-2 diabetes and fatty liver was also associated with lower TREC numbers. This population study suggests that obesity with enhanced inflammation is involved in aging of the human T-cell immune system. Given the fact that obesity increases the risk of numerous age-related diseases, attenuated immune competence is a possible mechanistic link between obesity and disease development among the elderly.

  19. Inverse associations between obesity indicators and thymic T-cell production levels in aging atomic-bomb survivors.

    Directory of Open Access Journals (Sweden)

    Kengo Yoshida

    Full Text Available Reduction of the naive T-cell population represents a deteriorating state in the immune system that occurs with advancing age. In animal model studies, obesity compromises the T-cell immune system as a result of enhanced adipogenesis in primary lymphoid organs and systemic inflammation. In this study, to test the hypothesis that obesity may contribute to the aging of human T-cell immunity, a thousand atomic-bomb survivors were examined for obesity status and ability to produce naive T cells, i.e., T-cell receptor excision circle (TREC numbers in CD4 and CD8 T cells. The number of TRECs showed a strong positive correlation with naive T cell numbers, and lower TREC numbers were associated with higher age. We found that the TREC number was inversely associated with levels of obesity indicators (BMI, hemoglobin A1c and serum CRP levels. Development of type-2 diabetes and fatty liver was also associated with lower TREC numbers. This population study suggests that obesity with enhanced inflammation is involved in aging of the human T-cell immune system. Given the fact that obesity increases the risk of numerous age-related diseases, attenuated immune competence is a possible mechanistic link between obesity and disease development among the elderly.

  20. pH-sensitivity of YFP provides an intracellular indicator of programmed cell death

    Directory of Open Access Journals (Sweden)

    Purcel Sydney B

    2010-11-01

    Full Text Available Abstract Background Programmed cell death (PCD is an essential process for the life cycle of all multicellular organisms. In higher plants however, relatively little is known about the cascade of genes and signalling molecules responsible for the initiation and execution of PCD. To aid with the discovery and analysis of plant PCD regulators, we have designed a novel cell death assay based on low cytosolic pH as a marker of PCD. Results The acidification that occurs in the cytosol during plant PCD was monitored by way of the extinction of YFP fluorescence at low pH. This fluorescence was recovered experimentally when bringing the intracellular pH back to 7, demonstrating that there was no protein degradation of YFP. Because it uses YFP, the assay is none-destructive, does not interfere with the PCD process and allows time-lapse studies to be carried out. In addition, changes of sub-cellular localisation can be visualised during PCD using the protein of interest fused to RFP. Coupled to a transient expression system, this pH-based assay can be used to functionally analyse genes involved in PCD, using point mutations or co-expressing PCD regulators. Transfecting mBAX and AtBI-1in onion epidermal cells showed that the pH shift is downstream of PCD suppression by AtBI-1. In addition, this method can be used to score PCD in tissues of stably transformed transgenic lines. As proof of principle, we show the example of YFP extinction during xylogenesis in Arabidopsis. This demonstrates that the assay is applicable to PCD studies in a variety of tissues. Conclusions The observation that YFP fluorescence is lost during the plant PCD process provides a new tool to study the genetic regulation and cell biology of the process. In addition, plant cell biologists should make a note of this effect of PCD on YFP fluorescence to avoid misinterpretation of their data and to select a pH insensitive reporter if appropriate. This method represents an efficient and

  1. Direct and indirect measurement of somatic cell count as indicator of intramammary infection in dairy goats.

    Science.gov (United States)

    Persson, Ylva; Olofsson, Ida

    2011-03-04

    Mastitis is the most important and costly disease in dairy goat production. Subclinical mastitis is common in goats and is mainly caused by contagious bacteria. Several methods to diagnose subclinical mastitis are available. In this study indirect measurement of somatic cell count (SCC) by California Mastitis Test (CMT) and direct measurement of SCC using a portable deLaval cell counter (DCC) are evaluated. Swedish goat farmers would primarily benefit from diagnostic methods that can be used at the farm. The purpose of the study was to evaluate SCC measured by CMT and DCC as possible markers for intramammary infection (IMI) in goats without clinical symptoms of mastitis. Moreover to see how well indirect measurement of SCC (CMT) corresponded to direct measurement of SCC (DCC). Udder half milk samples were collected once from dairy goats (n = 111), in five different farms in Northern and Central Sweden. Only clinically healthy animals were included in the study. All goats were in mid to late lactation at sampling. Milk samples were analyzed for SCC by CMT and DCC at the farm, and for bacterial growth at the laboratory. Intramammary infection, defined as growth of udder pathogens, was found in 39 (18%) of the milk samples. No growth was found in 180 (81%) samples while 3 (1%) samples were contaminated. The most frequently isolated bacterial species was coagulase negative staphylococci (CNS) (72% of all isolates), followed by Staphylococcus aureus (23% of all isolates). Somatic cell count measured by DCC was strongly (p = 0.000) associated with bacterial growth. There was also a very strong association between CMT and bacterial growth. CMT 1 was associated with freedom of IMI while CMT ≥2 was associated with IMI. Indirect measurement of SCC by CMT was well correlated with SCC measured by DCC. According to the results, SCC measured with CMT or DCC can predict udder infection in goats, and CMT can be used as a predictor of the SCC.

  2. Direct and indirect measurement of somatic cell count as indicator of intramammary infection in dairy goats

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    Olofsson Ida

    2011-03-01

    Full Text Available Abstract Background Mastitis is the most important and costly disease in dairy goat production. Subclinical mastitis is common in goats and is mainly caused by contagious bacteria. Several methods to diagnose subclinical mastitis are available. In this study indirect measurement of somatic cell count (SCC by California Mastitis Test (CMT and direct measurement of SCC using a portable deLaval cell counter (DCC are evaluated. Swedish goat farmers would primarily benefit from diagnostic methods that can be used at the farm. The purpose of the study was to evaluate SCC measured by CMT and DCC as possible markers for intramammary infection (IMI in goats without clinical symptoms of mastitis. Moreover to see how well indirect measurement of SCC (CMT corresponded to direct measurement of SCC (DCC. Method Udder half milk samples were collected once from dairy goats (n = 111, in five different farms in Northern and Central Sweden. Only clinically healthy animals were included in the study. All goats were in mid to late lactation at sampling. Milk samples were analyzed for SCC by CMT and DCC at the farm, and for bacterial growth at the laboratory. Results Intramammary infection, defined as growth of udder pathogens, was found in 39 (18% of the milk samples. No growth was found in 180 (81% samples while 3 (1% samples were contaminated. The most frequently isolated bacterial species was coagulase negative staphylococci (CNS (72% of all isolates, followed by Staphylococcus aureus (23% of all isolates. Somatic cell count measured by DCC was strongly (p = 0.000 associated with bacterial growth. There was also a very strong association between CMT and bacterial growth. CMT 1 was associated with freedom of IMI while CMT ≥2 was associated with IMI. Indirect measurement of SCC by CMT was well correlated with SCC measured by DCC. Conclusions According to the results, SCC measured with CMT or DCC can predict udder infection in goats, and CMT can be used as a

  3. Proteomics indicates modulation of tubulin polymerization by L-menthol inhibiting human epithelial colorectal adenocarcinoma cell proliferation.

    Science.gov (United States)

    Faridi, Uzma; Sisodia, Brijesh S; Shukla, Ashutosh K; Shukla, Rakesh K; Darokar, Mahendra P; Dwivedi, Upendra N; Shasany, Ajit K

    2011-05-01

    Menthol is a naturally occurring cyclic monoterpene used in oral hygiene products, confectionary, pharmaceuticals, cosmetics, pesticides, and as a flavoring agent. In the present study, we analyzed the differentially expressing proteome in L-menthol-treated Caco-2 cell line as it was found to inhibit cell proliferation. Interestingly, free tubulin proteins were observed to be limited after menthol treatment. Semiquantitative RT-PCR with α-tubulin primers showed no change in the level of RNA expression in menthol-treated cell line. However, tubulin polymerization assay with menthol indicated a trend similar to taxol in promoting microtubule assembly. Further, physical counting of apoptotic nuclei and active caspase-3 assays confirmed onset of apoptosis though the rate was slower as compared with that of taxol treatment. This study is the first report of a monoterpene L-menthol modulating tubulin polymerization and apoptosis to inhibit cancer cell proliferation.

  4. Thalidomide induced early gene expression perturbations indicative of human embryopathy in mouse embryonic stem cells.

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    Gao, Xiugong; Sprando, Robert L; Yourick, Jeffrey J

    2015-08-15

    Developmental toxicity testing has traditionally relied on animal models which are costly, time consuming, and require the sacrifice of large numbers of animals. In addition, there are significant disparities between human beings and animals in their responses to chemicals. Thalidomide is a species-specific developmental toxicant that causes severe limb malformations in humans but not in mice. Here, we used microarrays to study transcriptomic changes induced by thalidomide in an in vitro model based on differentiation of mouse embryonic stem cells (mESCs). C57BL/6 mESCs were allowed to differentiate spontaneously and RNA was collected at 24, 48, and 72h after exposure to 0.25mM thalidomide. Global gene expression analysis using microarrays revealed hundreds of differentially expressed genes upon thalidomide exposure that were enriched in gene ontology (GO) terms and canonical pathways associated with embryonic development and differentiation. In addition, many genes were found to be involved in small GTPases-mediated signal transduction, heart development, and inflammatory responses, which coincide with clinical evidences and may represent critical embryotoxicities of thalidomide. These results demonstrate that transcriptomics in combination with mouse embryonic stem cell differentiation is a promising alternative model for developmental toxicity assessment.

  5. Intrinsic indicator of photodamage during label-free multiphoton microscopy of cells and tissues.

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    Roberta Galli

    Full Text Available Multiphoton imaging has evolved as an indispensable tool in cell biology and holds prospects for clinical applications. When addressing endogenous signals such as coherent anti-Stokes Raman scattering (CARS or second harmonic generation, it requires intense laser irradiation that may cause photodamage. We report that increasing endogenous fluorescence signal upon multiphoton imaging constitutes a marker of photodamage. The effect was studied on mouse brain in vivo and ex vivo, on ex vivo human brain tissue samples, as well as on glioblastoma cells in vitro, demonstrating that this phenomenon is common to a variety of different systems, both ex vivo and in vivo. CARS microscopy and vibrational spectroscopy were used to analyze the photodamage. The development of a standard easy-to-use model that employs rehydrated cryosections allowed the characterization of the irradiation-induced fluorescence and related it to nonlinear photodamage. In conclusion, the monitoring of endogenous two-photon excited fluorescence during label-free multiphoton microscopy enables to estimate damage thresholds ex vivo as well as detect photodamage during in vivo experiments.

  6. Large-scale population study of human cell lines indicates that dosage compensation is virtually complete.

    Directory of Open Access Journals (Sweden)

    Colette M Johnston

    2008-01-01

    Full Text Available X chromosome inactivation in female mammals results in dosage compensation of X-linked gene products between the sexes. In humans there is evidence that a substantial proportion of genes escape from silencing. We have carried out a large-scale analysis of gene expression in lymphoblastoid cell lines from four human populations to determine the extent to which escape from X chromosome inactivation disrupts dosage compensation. We conclude that dosage compensation is virtually complete. Overall expression from the X chromosome is only slightly higher in females and can largely be accounted for by elevated female expression of approximately 5% of X-linked genes. We suggest that the potential contribution of escape from X chromosome inactivation to phenotypic differences between the sexes is more limited than previously believed.

  7. Multivariate analysis of cell culture bioprocess data--lactate consumption as process indicator.

    Science.gov (United States)

    Le, Huong; Kabbur, Santosh; Pollastrini, Luciano; Sun, Ziran; Mills, Keri; Johnson, Kevin; Karypis, George; Hu, Wei-Shou

    2012-12-31

    Multivariate analysis of cell culture bioprocess data has the potential of unveiling hidden process characteristics and providing new insights into factors affecting process performance. This study investigated the time-series data of 134 process parameters acquired throughout the inoculum train and the production bioreactors of 243 runs at the Genentech's Vacaville manufacturing facility. Two multivariate methods, kernel-based support vector regression (SVR) and partial least square regression (PLSR), were used to predict the final antibody concentration and the final lactate concentration. Both product titer and the final lactate level were shown to be predicted accurately when data from the early stages of the production scale were employed. Using only process data from the inoculum train, the prediction accuracy of the final process outcome was lower; the results nevertheless suggested that the history of the culture may exert significant influence on the final process outcome. The parameters contributing most significantly to the prediction accuracy were related to lactate metabolism and cell viability in both the production scale and the inoculum train. Lactate consumption, which occurred rather independently of the residual glucose and lactate concentrations, was shown to be a prominent factor in determining the final outcome of production-scale cultures. The results suggest possible opportunities to intervene in metabolism, steering it towards the type with a strong propensity towards high productivity. Such intervention could occur in the inoculum stage or in the early stage of the production-scale reactors. Overall, this study presents pattern recognition as an important process analytical technology (PAT). Furthermore, the high correlation between lactate consumption and high productivity can provide a guide to apply quality by design (QbD) principles to enhance process robustness.

  8. Structural and electrochemical studies of a hexa phenylbenzene pyro lysed soft carbon as anode material in lithium batteries

    Energy Technology Data Exchange (ETDEWEB)

    Bonino, Franco [Chemistry Department, University ' La Sapienza' , P. le A. Moro 5, 00185 Rome (Italy)]. E-mail: bonino@uniromal.it; Brutti, Sergio [Chemistry Department, University ' La Sapienza' , P. le A. Moro 5, 00185 Rome (Italy); Piana, Michele [Chemistry Department, University ' La Sapienza' , P. le A. Moro 5, 00185 Rome (Italy); Natale, Sergio [Chemistry Department, University ' La Sapienza' , P. le A. Moro 5, 00185 Rome (Italy); Scrosati, Bruno [Chemistry Department, University ' La Sapienza' , P. le A. Moro 5, 00185 Rome (Italy); Gherghel, Lileta [Max-Planck Institute for Polymer Research, Ackermannweg 10, 55124 Mainz (Germany); Muellen, Klaus [Max-Planck Institute for Polymer Research, Ackermannweg 10, 55124 Mainz (Germany)

    2006-04-25

    XRD, Sem micrographs, Bet analyses and typical electrochemical experiments (cyclic voltammetry, step voltammetry and Li insertion/de insertion at constant current) have been carried out to characterize a new type of soft carbons obtained by pyrolysis of hexa phenylbenzene (Hb). By means of XRD and cyclic voltammetry at least three different type of sites for lithium storage were found. The first is graphite like type with d {sub 002} graph ene layer distance greater than pure graphite; the second is associated to disordered volumes among crystallities and the third is represented by Li sites at the hydrogen-terminated edges of hexagonal carbon fragments, characterized by higher energy in comparison with simple insertion sites. These last two types of sites are able to store some extra lithium, compared to pure graphite. BET analyses and cyclic voltammetries demonstrate the key role of the milling time on the characteristics and properties of this HPB pyrolysed carbon. Specific capacities shown by this pyrolysed material in Li coin-type cell have been also reported.

  9. Hematopoietic stem cell transplantation for follicular lymphoma: optimal timing and indication.

    Science.gov (United States)

    Kim, Sung Won

    2014-01-01

    The definitive management of advanced follicular lymphoma (FL) remains controversial due to various treatment options, including watchful waiting, single-agent or combination chemotherapy, monoclonal antibody, and radioimmunotherapy. These options can provide prolonged progression-free survival. However, they cannot cure advanced FL. Allogeneic hematopoietic stem cell transplantation (allo-SCT) remains the sole curative therapy for FL. Allo-SCT has had a major impact with the use of reduced-intensity conditioning regimens because of its lower associated nonrelapse mortality compared with myeloablative regimens. Autologous SCT (auto-SCT) shows high response rates and extends progression-free survival in patients with chemosensitive relapse. In the rituximab era, however, associated comorbidities, risk of secondary cancers, and presence of refractory disease have become problematic in the auto-SCT population. On the basis of results from large-scale randomized trials, upfront auto-SCT is not recommended. Novel conditioning regimens including radioimmunotherapy followed by either auto-SCT or allo-SCT are likely to show efficacy even in chemorefractory disease. Consequently, the optimal timing for SCT remains a matter of opinion, except for patients in first remission. However, the outcomes of allo-SCT and auto-SCT keep on improving. Physicians should note that there is no therapy with a track record equivalent to that of SCT for relapsed or refractory FL.

  10. Extrapulmonary small cell carcinoma. An indication for prophylactic cranial irradiation? A single center experience

    Energy Technology Data Exchange (ETDEWEB)

    Frueh, Martin; Cerny, Thomas [Kantonsspital St. Gallen (Switzerland). Dept. of Oncology/Hematology; Kacsir, Bela; Plasswilm, Ludwig [Kantonsspital St. Gallen (Switzerland). Dept. of Radiation Oncology; Ess, Silvia [Cancer Registry, Canton of St. Gallen, Appenzell (Switzerland); Rodriguez, Regulo [Kantonsspital St. Gallen (Switzerland). Dept. of Pathology

    2011-09-15

    Information about extrapulmonary small cell carcinoma (EPSCC) is limited and the role of prophylactic cranial irradiation (PCI) is unknown. Disease presentation and outcome of all EPSCC at our hospital between 1990 and 2009 were retrospectively analyzed. Of 30 EPSCC, the male:female ratio was 58%:42%; 83% had a performance status of 0-2. Median age was 71 years (32-80). Seventeen (57%) had limited stage (LS), 13 (43%) extensive stage (ES). The location of the primary tumor was gastrointestinal (n = 8), unknown (6), gynecological (6), urogenital (5), and ear nose throat (5). Four (13%) developed brain metastases (2 ES, 2 LS). In ES, first line chemotherapy (CT) was given in 85%, mostly platinum-etoposide (64%). Response rate was 90%. In LS, CT and radiotherapy (RT) {+-} resection resulted in persistent remissions in 67% of patients. Median survival was 16 months (1-107 months), 18 months (1-107 months), and 9 months (0.4-25 months) for LS + ES, LS, and ES, respectively. Weight loss {>=}5 % and ECOG performance status 3 + 4 were associated with poorer survival (p < 0.001 and p < 0.01, respectively). The incidence of brain metastases was relatively low (13%). More studies are necessary, before routinely offering PCI to patients with EPSCC. Best survival outcomes in LS were achieved with multimodality treatment including CT and RT. Prognosis was poor in patients with ES. (orig.)

  11. Stromal fibers in oral squamous cell carcinoma: A possible new prognostic indicator?

    Science.gov (United States)

    Kardam, Priyanka; Mehendiratta, Monica; Rehani, Shweta; Kumra, Madhumani; Sahay, Khushboo; Jain, Kanu

    2016-01-01

    Background: Many studies have been carried out to study the role of extracellular matrix proteins, growth factors and matrix metalloproteinases on tumor invasion. However, literature related to the analysis of connective tissue fibers in varying grades of oral squamous cell carcinoma (OSCC) is very limited. Aim: To analyze the changes in collagen and elastic fibers in varying grades of (OSCC). Settings and Design: This retrospective study was carried out using a light and polarizing microscope. Materials and Methods: Three sections each were cut from fifty samples of varying grades of OSCC and ten samples of control followed by staining with H and E, Picrosirius-Red and Verhoeff–Van Gieson. Qualitative and quantitative analysis of collagen and elastic fibers were accomplished using set criteria. Statistical Analysis: Data were entered into the Statistical Package for Social Sciences (SPSS) version 13.5 for analysis. Results: A change in colors of collagen fibers was seen on progressing from well to poorly differentiated OSCC. Thin collagen fibers predominantly exhibited greenish yellow, but the thick fibers exhibited a variety of colors. As the grade of OSCC progressed, collagen fibers were loosely packed haphazardly arranged. Statistically insignificant results were obtained for quantitative analysis of collagen and qualitative analysis of elastic fibers. Conclusion: The collagen fibers undergo a change in color, orientation and packing in the stroma of varying grades of OSCC. The uniqueness of this study lies in the exploration of elastic fibers in OSCC which has not been done so far. PMID:27721605

  12. Rapid 96-well plates DNA extraction and sequencing procedures to identify genome-wide transposon insertion sites in a difficult to lyse bacterium: Lactobacillus casei.

    Science.gov (United States)

    Scornec, Hélène; Tichit, Magali; Bouchier, Christiane; Pédron, Thierry; Cavin, Jean-François; Sansonetti, Philippe J; Licandro-Seraut, Hélène

    2014-11-01

    Random transposon mutagenesis followed by adequate screening methods is an unavoidable procedure to characterize genetics of bacterial adaptation to environmental changes. We have recently constructed a mutant library of Lactobacillus casei and we aimed to fully annotate it. However, we have observed that, for L. casei which is a difficult to lyse bacterium, methods used to identify the transposon insertion site in a few mutants (transposon rescue by restriction and recircularization or PCR-based methods) were not transposable for a larger number because they are too time-consuming and sometimes not reliable. Here, we describe a method for large-scale and reliable identification of transposon insertion sites in a L. casei mutant library of 9250 mutants. DNA extraction procedure based on silica membranes in 96-column format was optimized to obtain genomic DNA from a large number of mutants. Then reliable direct genomic sequencing was improved to fit the obtained genomic DNA extracts. Using this procedure, readable and identifiable sequences were obtained for 87% of the L. casei mutants. This method extends the applications of a library of this type, reduces the number of insertions needed to be screened, and allows selection of specific mutants from an arrayed and stored mutant library. This method is applicable to any already existing mutant library (obtained by transposon or insertional mutagenesis) and could be useful for other bacterial species, especially for highly lysis-resistant bacteria species such as lactic acid bacteria. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Type II cytokeratin gene expression is indicative of early cell differentiation in the chick embryo

    Energy Technology Data Exchange (ETDEWEB)

    Charlebois, T.S.

    1988-01-01

    Embryonic development in vertebrates appears to involve a series of inductive tissue interactions that lead to regional specializations, which eventually become elaborated in the basic body plan of the embryo. The inductive interactions leading to early regionalization of the embryo are often particularly difficult to evaluate because of the absence of available morphological or biochemical evidence that such events have occurred. In the 36 hour chick embryo, the regional subdivision of the early ectoderm is evidence by a marked lens-forming bias in the head ectoderm, which is absent in the presumptive dorsal epidermis of the trunk region. As a strategy for isolating genes whose differential expression might reflect this regional subdivision, a cDNA library from 36 hour embryos was prepared and screened for differential hybridization to ({sup 32}P)cDNA probes synthesized using template RNA isolated from 36 hour head ectoderm and trunk ectoderm. A cDNA clone (T4) was isolated which hybridizes to transcripts present at much higher levels in trunk ectoderm than in head ectoderm. Partial nucleotide and deduced amino acid sequences of this clone indicate that it represents a gene encoding a type II cytokeratin. The distribution of transcripts complementary to the T4 probe was evaluated in early embryos using RNA gel blot analysis and in situ hybridization to tissue sections.

  14. Phylogenomic Analyses Indicate that Early Fungi Evolved Digesting Cell Walls of Algal Ancestors of Land Plants

    Science.gov (United States)

    Chang, Ying; Wang, Sishuo; Sekimoto, Satoshi; Aerts, Andrea L.; Choi, Cindy; Clum, Alicia; LaButti, Kurt M.; Lindquist, Erika A.; Yee Ngan, Chew; Ohm, Robin A.; Salamov, Asaf A.; Grigoriev, Igor V.; Spatafora, Joseph W.; Berbee, Mary L.

    2015-01-01

    As decomposers, fungi are key players in recycling plant material in global carbon cycles. We hypothesized that genomes of early diverging fungi may have inherited pectinases from an ancestral species that had been able to extract nutrients from pectin-containing land plants and their algal allies (Streptophytes). We aimed to infer, based on pectinase gene expansions and on the organismal phylogeny, the geological timing of the plant–fungus association. We analyzed 40 fungal genomes, three of which, including Gonapodya prolifera, were sequenced for this study. In the organismal phylogeny from 136 housekeeping loci, Rozella diverged first from all other fungi. Gonapodya prolifera was included among the flagellated, predominantly aquatic fungal species in Chytridiomycota. Sister to Chytridiomycota were the predominantly terrestrial fungi including zygomycota I and zygomycota II, along with the ascomycetes and basidiomycetes that comprise Dikarya. The Gonapodya genome has 27 genes representing five of the seven classes of pectin-specific enzymes known from fungi. Most of these share a common ancestry with pectinases from Dikarya. Indicating functional and sequence similarity, Gonapodya, like many Dikarya, can use pectin as a carbon source for growth in pure culture. Shared pectinases of Dikarya and Gonapodya provide evidence that even ancient aquatic fungi had adapted to extract nutrients from the plants in the green lineage. This implies that 750 million years, the estimated maximum age of origin of the pectin-containing streptophytes represents a maximum age for the divergence of Chytridiomycota from the lineage including Dikarya. PMID:25977457

  15. Phylogenomic Analyses Indicate that Early Fungi Evolved Digesting Cell Walls of Algal Ancestors of Land Plants.

    Science.gov (United States)

    Chang, Ying; Wang, Sishuo; Sekimoto, Satoshi; Aerts, Andrea L; Choi, Cindy; Clum, Alicia; LaButti, Kurt M; Lindquist, Erika A; Yee Ngan, Chew; Ohm, Robin A; Salamov, Asaf A; Grigoriev, Igor V; Spatafora, Joseph W; Berbee, Mary L

    2015-05-14

    As decomposers, fungi are key players in recycling plant material in global carbon cycles. We hypothesized that genomes of early diverging fungi may have inherited pectinases from an ancestral species that had been able to extract nutrients from pectin-containing land plants and their algal allies (Streptophytes). We aimed to infer, based on pectinase gene expansions and on the organismal phylogeny, the geological timing of the plant-fungus association. We analyzed 40 fungal genomes, three of which, including Gonapodya prolifera, were sequenced for this study. In the organismal phylogeny from 136 housekeeping loci, Rozella diverged first from all other fungi. Gonapodya prolifera was included among the flagellated, predominantly aquatic fungal species in Chytridiomycota. Sister to Chytridiomycota were the predominantly terrestrial fungi including zygomycota I and zygomycota II, along with the ascomycetes and basidiomycetes that comprise Dikarya. The Gonapodya genome has 27 genes representing five of the seven classes of pectin-specific enzymes known from fungi. Most of these share a common ancestry with pectinases from Dikarya. Indicating functional and sequence similarity, Gonapodya, like many Dikarya, can use pectin as a carbon source for growth in pure culture. Shared pectinases of Dikarya and Gonapodya provide evidence that even ancient aquatic fungi had adapted to extract nutrients from the plants in the green lineage. This implies that 750 million years, the estimated maximum age of origin of the pectin-containing streptophytes represents a maximum age for the divergence of Chytridiomycota from the lineage including Dikarya.

  16. Isolation of a T-cell clone showing HLA-DRB1*0405-restricted cytotoxicity for hematopoietic cells in a patient with aplastic anemia.

    Science.gov (United States)

    Nakao, S; Takami, A; Takamatsu, H; Zeng, W; Sugimori, N; Yamazaki, H; Miura, Y; Ueda, M; Shiobara, S; Yoshioka, T; Kaneshige, T; Yasukawa, M; Matsuda, T

    1997-05-15

    The existence of T cells capable of inhibiting in vitro hematopoiesis has been shown in aplastic anemia (AA), although whether such inhibition is mediated by a specific immune reaction involving an HLA allele remained unknown. We isolated a CD4+ Vbeta21+ T-cell clone that was most dominant among Vbeta21+ T cells in the bone marrow (BM) of an AA patient whose HLA-DRB1 alleles included 1501 and 0405. The T-cell clone named NT4.2 lysed an autologous Epstein-Barr virus-transformed lymphoblastoid cell line (LCL) and phytohemagglutinin-stimulated lymphocytes (PHA-blasts) as well as allogeneic LCLs sharing HLA-DRB1*0405. Cytotoxicity against LCL cells and PHA-blasts by NT4.2 was blocked by anti-HLA-DR monoclonal antibody (MoAb) or anti-CD3 MoAb. NT4.2 also lysed autologous BM mononuclear cells enriched with CD34+ cells that had been cultured for one week in the presence of colony-stimulating factors as well as allogeneic CD34+ cells of a normal individual carrying HLA-DRB1*0405, cultured in the same way. Moreover, NT4.2 strongly inhibited colony formation by hematopoietic progenitor cells derived from cultured CD34+ cells sharing HLA-DRB1*0405. These results indicate that the AA patient has T cells capable of killing hematopoietic cells in an HLA-DRB1*0405-restricted manner and that such cytotoxic T cells may contribute to the pathogenesis of AA.

  17. Single Entity Electrochemistry Progresses to Cell Counting.

    Science.gov (United States)

    Gooding, J Justin

    2016-10-10

    Red blood cells have been counted in an electrochemical collision experiment recently described by Compton and co-workers. As a cell collides with the electrode it lyses and a current is observed from the reduction of oxygen from within the cell.

  18. Real-time visualization of calcium ion activity in shallow benthic foraminiferal cells using the fluorescent indicator Fluo-3 AM

    Science.gov (United States)

    Toyofuku, Takashi; Jan de Nooijer, Lennart; Yamamoto, Hiroyuki; Kitazato, Hiroshi

    2008-05-01

    Calcium ion storage and movements within foraminiferal cells were observed using the fluorescent cell-permeant calcium indicator Fluo-3 AM. Living specimens of the shallow-water benthic foraminifer Ammonia beccarii were incubated with 8 μM Fluo-3 AM in both natural seawater and calcium-free artificial seawater. No fluorescence was observed in specimens that were incubated in Fluo-3 AM/calcium-free seawater, while fluorescent emission was identified in all foraminifera that were incubated in Fluo-3 AM/natural seawater. In another series of experiments, juvenile specimens were incubated with Fluo-3 AM to trace calcium ion activity during the process of chamber formation. The method described here is a potential tool to observe the flow of calcium ions within living foraminiferal cells and may yield valuable information on the process of calcite precipitation.

  19. Auto-luminescent genetically-encoded ratiometric indicator for real-time Ca2+ imaging at the single cell level.

    Directory of Open Access Journals (Sweden)

    Kenta Saito

    Full Text Available BACKGROUND: Efficient bioluminescence resonance energy transfer (BRET from a bioluminescent protein to a fluorescent protein with high fluorescent quantum yield has been utilized to enhance luminescence intensity, allowing single-cell imaging in near real time without external light illumination. METHODOLOGY/PRINCIPAL FINDINGS: We applied BRET to develop an autoluminescent Ca(2+ indicator, BRAC, which is composed of Ca(2+-binding protein, calmodulin, and its target peptide, M13, sandwiched between a yellow fluorescent protein variant, Venus, and an enhanced Renilla luciferase, RLuc8. Adjusting the relative dipole orientation of the luminescent protein's chromophores improved the dynamic range of BRET signal change in BRAC up to 60%, which is the largest dynamic range among BRET-based indicators reported so far. Using BRAC, we demonstrated successful visualization of Ca(2+ dynamics at the single-cell level with temporal resolution at 1 Hz. Moreover, BRAC signals were acquired by ratiometric imaging capable of canceling out Ca(2+-independent signal drifts due to change in cell shape, focus shift, etc. CONCLUSIONS/SIGNIFICANCE: The brightness and large dynamic range of BRAC should facilitate high-sensitive Ca(2+ imaging not only in single live cells but also in small living subjects.

  20. Red Cell Indices and Formulas Used in Differentiation of β-Thalassemia Trait from Iron Deficiency in Thai Adults.

    Science.gov (United States)

    Plengsuree, Soontharee; Punyamung, Manoo; Yanola, Jintana; Nanta, Siriporn; Jaiping, Kanokwan; Maneewong, Kritsanee; Wongwiwatthananukit, Sanchai; Pornprasert, Sakorn

    2015-01-01

    β-Thalassemia (β-thal) and iron deficiency cause most microcytic anemias. Red cell indices and formulas have been established as simple, fast, and inexpensive in discrimination between these two hematological disorders in school children. However, whether these formulas could be applied to diagnose β-thal trait and iron deficiency in adult Thai subjects is unclear. The aim of this study was to examine the diagnostic accuracy of five red cell indices [red blood cell (RBC) counts, mean corpuscular volume (MCV), mean corpuscular hemoglobin (Hb) (MCH), mean corpuscular Hb concentration (MCHC), and red cell distribution width (RDW)] and nine formulas (RDW/RBC, RDW Index, Sirdah, Green and King, Mentzer, England and Fraser, Ehsani, Srivastava and Shine and Lal). Their sensitivity, specificity, positive predictive value (PPV), and negative predictive values (NPV), efficiency, and Youden's Index were analyzed in 102 β-thal trait and 64 iron deficiency adult Thai subjects. The RDW/RBC formula proved to be the most reliable index as they had 100.0% specificity and PPV and the highest efficiency (94.58%) and Youden's Index (91.18%), as well as high sensitivity (91.18%) and NPV (87.67%). Therefore, this formula could be used in initial discrimination of β-thal trait from iron deficiency in adult Thai subjects.

  1. Conserved B-cell epitopes among human bocavirus species indicate potential diagnostic targets.

    Directory of Open Access Journals (Sweden)

    Zhuo Zhou

    Full Text Available BACKGROUND: Human bocavirus species 1-4 (HBoV1-4 have been associated with respiratory and enteric infections in children. However, the immunological mechanisms in response to HBoV infections are not fully understood. Though previous studies have shown cross-reactivities between HBoV species, the epitopes responsible for this phenomenon remain unknown. In this study, we used genomic and immunologic approaches to identify the reactive epitopes conserved across multiple HBoV species and explored their potential as the basis of a novel diagnostic test for HBoVs. METHODOLOGY/PRINCIPAL FINDINGS: We generated HBoV1-3 VP2 gene fragment phage display libraries (GFPDLs and used these libraries to analyze mouse antisera against VP2 protein of HBoV1, 2, and 3, and human sera positive for HBoVs. Using this approach, we mapped four epitope clusters of HBoVs and identified two immunodominant peptides--P1 (¹MSDTDIQDQQPDTVDAPQNT²⁰, and P2 (¹⁶²EHAYPNASHPWDEDVMPDL¹⁸⁰--that are conserved among HBoV1-4. To confirm epitope immunogenicity, we immunized mice with the immunodominant P1 and P2 peptides identified in our screen and found that they elicited high titer antibodies in mice. These two antibodies could only recognize the VP2 of HBoV 1-4 in Western blot assays, rather than those of the two other parvoviruses human parvovirus B19 and human parvovirus 4 (PARV4. Based on our findings, we evaluated epitope-based peptide-IgM ELISAs as potential diagnostic tools for HBoVs IgM antibodies. We found that the P1+P2-IgM ELISA showed a higher sensitivity and specificity in HBoVs IgM detection than the assays using a single peptide. CONCLUSIONS/SIGNIFICANCE: The identification of the conserved B-cell epitopes among human bocavirus species contributes to our understanding of immunological cross-reactivities of HBoVs, and provides important insights for the development of HBoV diagnostic tools.

  2. Vitamin D₃ and monomethyl fumarate enhance natural killer cell lysis of dendritic cells and ameliorate the clinical score in mice suffering from experimental autoimmune encephalomyelitis.

    Science.gov (United States)

    Al-Jaderi, Zaidoon; Maghazachi, Azzam A

    2015-11-13

    Experimental autoimmune encephalomyelitis (EAE) is a CD4⁺ T cell mediated inflammatory demyelinating disease that is induced in mice by administration of peptides derived from myelin proteins. We developed EAE in SJL mice by administration of PLP139-151 peptide. The effect of treating these mice with 1α,25-Dihydroxyvitamin D₃ (vitamin D₃), or with monomethyl fumarate (MMF) was then examined. We observed that both vitamin D₃ and MMF inhibited and/or prevented EAE in these mice. These findings were corroborated with isolating natural killer (NK) cells from vitamin D₃-treated or MMF-treated EAE mice that lysed immature or mature dendritic cells. The results support and extend other findings indicating that an important mechanism of action for drugs used to treat multiple sclerosis (MS) is to enhance NK cell lysis of dendritic cells.

  3. Single Cell Characterization of Prostate Cancer-Circulating Tumor Cells

    Science.gov (United States)

    2013-09-01

    al., 2010). In addition, there were a significant number of cell cycle and mitosis associated transcripts in the highly expressed gene set including...red blood cell lysis with 10 volumes of 16 PharmLyse (BD Biosciences) for 15 minutes at room temperature . Remaining cells were pelleted at 4uC for 15...processes (23%, GO:0008152) or the cell cycle (10%, GO:0007049), consistent with mitotically active cells (Fig. 4C). Cell cycle and mitosis associated

  4. Ratiometric bioluminescence indicators for monitoring cyclic adenosine 3',5'-monophosphate in live cells based on luciferase-fragment complementation.

    Science.gov (United States)

    Takeuchi, Masaki; Nagaoka, Yasutaka; Yamada, Toshimichi; Takakura, Hideo; Ozawa, Takeaki

    2010-11-15

    Bioluminescent indicators for cyclic 3',5'-monophosphate AMP (cAMP) are powerful tools for noninvasive detection with high sensitivity. However, the absolute photon counts are affected substantially by adenosine 5'-triphosphate (ATP) and d-luciferin concentrations, limiting temporal analysis in live cells. This report describes a genetically encoded bioluminescent indicator for detecting intracellular cAMP based on complementation of split fragments of two-color luciferase mutants originated from click beetles. A cAMP binding domain of protein kinase A was connected with an engineered carboxy-terminal fragment of luciferase, of which ends were connected with amino-terminal fragments of green luciferase and red luciferase. We demonstrated that the ratio of green to red bioluminescence intensities was less influenced by the changes of ATP and d-luciferin concentrations. We also showed an applicability of the bioluminescent indicator for time-course and quantitative assessments of intracellular cAMP in living cells and mice. The bioluminescent indicator will enable quantitative analysis and imaging of spatiotemporal dynamics of cAMP in opaque and autofluorescent living subjects.

  5. Basal cytokeratin phenotypes of myoepithelial cells indicates the origin of ductal carcinomas in situ of the breast.

    Science.gov (United States)

    Chen, Ling; Yin, Xiaona; Lu, Shanshan; Chen, Guorong; Dong, Lei

    2015-09-01

    Terminal duct lobular unit (TDLU) is widely accepted as the origin of ductal carcinoma in situ of breast. The differentiation states of myoepithelial cells of breast ductal system hint the development of breast hyperplastic lesions. Basal cytokeratin (CK) phenotypes indicate the differentiation of myoepithelial cells. Using antibodies of CK5/6, CK14, and CK17, this study reports the basal CK phenotypes of myoepithelial cells in 20 foci of normal breast, 20 usual ductal hyperplasias, 36 ductal carcinomas in situ (DCIS), and 28 sclerosing adenosis (SA). The results showed that the positive staining of basal CKs of myoepithelial cells in normal ducts were significantly higher than those in normal lobules. The basal CK expression of myoepithelial cells of DCIS and usual ductal hyperplasia was similar to that of normal duct, whereas that of SA was similar to that of normal lobule. We propose a modified model of TDLU origin of intraductal carcinoma that most of DCIS originate from terminal ducts of TDLU, whereas most SA originate from lobules.

  6. Indications of hematopoietic stem cell transplantations and therapeutic strategies of accidental irradiations; Indications des greffes de cellules souches hematopoietiques et strategies therapeutiques des irradiations accidentelles

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2003-07-01

    Produced by a group of experts, this document first discusses the issue of accidental irradiations in terms of medical management. They notably outline the peculiar characteristics of these irradiations with respect to therapeutic irradiations. They agreed on general principles regarding casualty sorting criteria and process, and their medical treatment (systematic hematopoiesis stimulation, allogeneic transplantation of hematopoietic stem cells). They discuss some practical aspects of these issues: casualty sorting within a therapeutic perspective (actions to be performed within 48 hours), therapeutic strategies (support therapy, use of cytokines, and therapy by hematopoietic stem cell transplant). They state a set of recommendations regarding the taking into care and diagnosis, therapeutic strategies, research perspectives, and teaching

  7. In HepG2 Cells, Coexisting Carnitine Deficiency Masks Important Indicators of Marginal Biotin Deficiency123

    Science.gov (United States)

    Bogusiewicz, Anna; Boysen, Gunnar; Mock, Donald M

    2015-01-01

    Background: A large number of birth defects are related to nutrient deficiencies; concern that biotin deficiency is teratogenic in humans is reasonable. Surprisingly, studies indicate that increased urinary 3-hydroxyisovalerylcarnitine (3HIAc), a previously validated marker of biotin deficiency, is not a valid biomarker in pregnancy. Objective: In this study we hypothesized that coexisting carnitine deficiency can prevent the increase in 3HIAc due to biotin deficiency. Methods: We used a 2-factor nutrient depletion design to induce isolated and combined biotin and carnitine deficiency in HepG2 cells and then repleted cells with carnitine. To elucidate the metabolic pathogenesis, we quantitated intracellular and extracellular free carnitine, acylcarnitines, and acylcarnitine ratios using liquid chromatography–tandem mass spectrometry. Results: Relative to biotin-sufficient, carnitine-sufficient cells, intracellular acetylcarnitine increased by 90%, propionylcarnitine more than doubled, and 3HIAc increased by >10-fold in biotin-deficient, carnitine-sufficient (BDCS) cells, consistent with a defensive mechanism in which biotin-deficient cells transesterify the acyl-coenzyme A (acyl-CoA) substrates of the biotin-dependent carboxylases to the related acylcarnitines. Likewise, in BDCS cells, the ratio of acetylcarnitine to malonylcarnitine and the ratio of propionylcarnitine to methylmalonylcarnitine both more than tripled, and the ratio of 3HIAc to 3-methylglutarylcarnitine (MGc) increased by >10-fold. In biotin-deficient, carnitine-deficient (BDCD) cells, the 3 substrate-derived acylcarnitines changed little, but the substrate:product ratios were masked to a lesser extent. Moreover, carnitine repletion unmasked biotin deficiency in BDCD cells as shown by increases in acetylcarnitine, propionylcarnitine, and 3HIAc (each increased by >50-fold). Likewise, ratios of acetylcarnitine:malonylcarnitine, propionylcarnitine:methylmalonylcarnitine, and 3HIAc:MGc all increased

  8. Differential mitochondrial calcium responses in different cell types detected with a mitochondrial calcium fluorescent indicator, mito-GCaMP2

    Institute of Scientific and Technical Information of China (English)

    Min Chen; Yanru Wang; Tingting Hou; Huiliang Zhang; Aijuan Qu; Xianhua Wang

    2011-01-01

    Mitochondrial calcium plays a crucial role in mitochondriai metabolism,cell calcium handling,and cell death.However,some mechanisms concerning mitochondrial calcium regulation are still unknown,especially how mitochondrial calcium couples with cytosolic calcium.In this work,we constructed a novel mitochondrial calcium fluorescent indicator (mito-GCaMP2) by genetic manipulation.Mito-GCaMP2 was imported into mitochondria with high efficiency and the fluorescent signals co-localized with that of tetramethyl rhodamine methyl ester,a mitochondrial membrane potential indicator.The mitochondrial inhibitors specifically decreased the signals of mito-GCaMP2.The apparent Kd of mito-GCaMP2 was 195.0 nmol/L at pH 8.0 in adult rat cardiomyocytes.Furthermore,we observed that mito-GCaMP2 preferred the alkaline pH surrounding of mitochondria.In HeLa cells,we found that mitochondrial calcium ([Ca2+]mito)responded to the changes of cytosolic calcium ([Ca2+]cyto)induced by histamine or thapasigargin.Moreover,external Ca2+ (100 μmol/L) directly induced an increase of [Ca2+]mito in permeabilized HeLa cells.However,in rat cardiomyocytes [Ca2+]mito did not respond to cytosolic calcium transients stimulated by electric pacing or caffeine.In permeabilized cardiomyocytes,600 nmol/L free Ca2+ repeatedly increased the fluorescent signals of mito-GCaMP2,which excluded the possibility that mito-GCaMP2 lost its function in cardiomyocytes mitochondria.These results showed that the response of mitochondrial calcium is diverse in different cell lineages and suggested that mitochondria in cardiomyocytes may have a special defense mechanism to control calcium flux.

  9. Long non-coding RNAs as surrogate indicators for chemical stress responses in human-induced pluripotent stem cells.

    Science.gov (United States)

    Tani, Hidenori; Onuma, Yasuko; Ito, Yuzuru; Torimura, Masaki

    2014-01-01

    In this study, we focused on two biological products as ideal tools for toxicological assessment: long non-coding RNAs (lncRNAs) and human-induced pluripotent stem cells (hiPSCs). lncRNAs are an important class of pervasive non-protein-coding transcripts involved in the molecular mechanisms associated with responses to cellular stresses. hiPSCs possess the capabilities of self-renewal and differentiation into multiple cell types, and they are free of the ethical issues associated with human embryonic stem cells. Here, we identified six novel lncRNAs (CDKN2B-AS1, MIR22HG, GABPB1-AS1, FLJ33630, LINC00152, and LINC0541471_v2) that respond to model chemical stresses (cycloheximide, hydrogen peroxide, cadmium, or arsenic) in hiPSCs. Our results indicated that the lncRNAs responded to general and specific chemical stresses. Compared with typical mRNAs such as p53-related mRNAs, the lncRNAs highly and rapidly responded to chemical stresses. We propose that these lncRNAs have the potential to be surrogate indicators of chemical stress responses in hiPSCs.

  10. Effect of Aloe vera on some indicators of cell damage after a period of aerobic exercise in male athletes

    Directory of Open Access Journals (Sweden)

    Barati Amir-Hosein

    2015-07-01

    Full Text Available Introduction: Aloe vera is a medicinal plant with antioxidant activity. The purpose of this study was to determine the effect of Aloe vera on some indicators of cell damage after a period of aerobic exercise in male athletes of 15-18 years old. Methods: In this clinical trial study, 20 male students with average weight of 64.85 ± 51 and height of 172.05 ± 6.4 were randomly assigned to 2 groups of Aloe vera supplement (n = 10 and placebo (n = 10. Then aerobic training was conducted for 4 weeks. The supplemented group took 3 capsules, each capsule contains 2 g of dried Aloe vera and placebo group took 3 capsules/day containing dextran after every meal. To determine the index of cell injury markers creatine kinase (CK, lactate dehydrogenase (LDH and reactive protein C (CRP were determined and blood samples were collected 24 hours before and after each test. The data were analyzed using ANOVA and independent t test. Significance level was considered as P ≤ .05. Results: The use of Aloe vera during aerobic exercise significantly reduced LDH (P = .006 in the supplement group (15% reduction compared to placebo group and CRP with 11% reduction in the supplement group compared to placebo (P = .008. There was no significant reduction in CK. Conclusion: The findings of this study showed that Aloe vera reduces cell damage and inflammation indicators. This result may reflect the role of Aloe vera as anti-inflammatory and antioxidant agent.

  11. Long non-coding RNAs as surrogate indicators for chemical stress responses in human-induced pluripotent stem cells.

    Directory of Open Access Journals (Sweden)

    Hidenori Tani

    Full Text Available In this study, we focused on two biological products as ideal tools for toxicological assessment: long non-coding RNAs (lncRNAs and human-induced pluripotent stem cells (hiPSCs. lncRNAs are an important class of pervasive non-protein-coding transcripts involved in the molecular mechanisms associated with responses to cellular stresses. hiPSCs possess the capabilities of self-renewal and differentiation into multiple cell types, and they are free of the ethical issues associated with human embryonic stem cells. Here, we identified six novel lncRNAs (CDKN2B-AS1, MIR22HG, GABPB1-AS1, FLJ33630, LINC00152, and LINC0541471_v2 that respond to model chemical stresses (cycloheximide, hydrogen peroxide, cadmium, or arsenic in hiPSCs. Our results indicated that the lncRNAs responded to general and specific chemical stresses. Compared with typical mRNAs such as p53-related mRNAs, the lncRNAs highly and rapidly responded to chemical stresses. We propose that these lncRNAs have the potential to be surrogate indicators of chemical stress responses in hiPSCs.

  12. Asymmetric localization of Vangl2 and Fz3 indicate novel mechanisms for planar cell polarity in mammals.

    Science.gov (United States)

    Montcouquiol, Mireille; Sans, Nathalie; Huss, David; Kach, Jacob; Dickman, J David; Forge, Andrew; Rachel, Rivka A; Copeland, Neal G; Jenkins, Nancy A; Bogani, Debora; Murdoch, Jennifer; Warchol, Mark E; Wenthold, Robert J; Kelley, Matthew W

    2006-05-10

    Planar cell polarity (PCP) is a process in which cells develop with uniform orientation within the plane of an epithelium. To begin to elucidate the mechanisms of PCP in vertebrates, the localization of the protein Vangl2 (Van Gogh-like) was determined during the development of the mammalian cochlea. Results indicate that Vangl2 becomes asymmetrically localized to specific cell-cell boundaries along the axis of polarization and that this asymmetry is lost in PCP mutants. In addition, PDZ2 (postsynaptic density/Discs large/zona occludens 1), PDZ3, and PDZ4 of the PCP protein Scrb1 (Scribble) are shown to bind to the C-terminal PDZ binding domain of Vangl2, suggesting that Scrb1 plays a direct role in asymmetric targeting of Vangl2. Finally, Fz3 (Frizzled), a newly demonstrated mediator of PCP, is also asymmetrically localized in a pattern that matches that of Vangl2. The presence and asymmetry of Fz3 at the membrane is shown to be dependent on Vangl2. This result suggests a role for Vangl2 in the targeting or anchoring of Fz3, a hypothesis strengthened by the existence of a physical interaction between the two proteins. Together, our data support the idea that protein asymmetry plays an important role in the development of PCP, but the colocalization and interaction of Fz3 and Vangl2 suggests that novel PCP mechanisms exist in vertebrates.

  13. Cobalt distribution in keratinocyte cells indicates nuclear and peri-nuclear accumulation and interaction with magnesium and zinc homeostasis

    Energy Technology Data Exchange (ETDEWEB)

    Ortega, R.; Seznec, H.; Moretto, Ph. [Univ Bordeaux 1, CNRS, IN2P3, Ctr Etud Nucl Bordeaux Gradignan, F-33175 Gradignan, (France); Bresson, C.; Sandre, C.; Gombert, C.; Moulin, Ch. [CEA, DEN, SECR, Lab Speciat Radionucleides et Mol, F-91191 Gif Sur Yvette, (France); Tabarant, M. [CEA, DEN, SCP, Lab React Surfaces et Interfaces, F-91191 Gif Sur Yvette, (France); Bleuet, P. [European Synchrotron Radiat Facil, F-38043 Grenoble, (France); Simionovici, A. [Univ Grenoble 1, LGIT, OSUG, CNRS, F-38041 Grenoble, (France)

    2009-07-01

    Cobalt is known to be toxic at high concentration, to induce contact dermatosis, and occupational radiation skin damage because of its use in nuclear industry. We investigated the intracellular distribution of cobalt in HaCaT human keratinocytes as a model of skin cells, and its interaction with endogenous trace elements. Direct micro-chemical imaging based on ion beam techniques was applied to determine the quantitative distribution of cobalt in HaCaT cells. In addition, synchrotron radiation X-ray fluorescence microanalysis in tomography mode was performed, for the first time on a single cell, to determine the 3D intracellular distribution of cobalt. Results obtained with these micro-chemical techniques were compared to a more classical method based on cellular fractionation followed by inductively coupled plasma atomic emission spectrometry (ICP-AES) measurements. Cobalt was found to accumulate in the cell nucleus and in peri-nuclear structures indicating the possible direct interaction with genomic DNA, and nuclear proteins. The peri-nuclear accumulation in the cytosol suggests that cobalt could be stored in the endoplasmic reticulum or the Golgi apparatus. The multi-elemental analysis revealed that cobalt exposure significantly decreased magnesium and zinc content, with a likely competition of cobalt for magnesium and zinc binding sites in proteins. Overall, these data suggest a multiform toxicity of cobalt related to interactions with genomic DNA and nuclear proteins, and to the alteration of zinc and magnesium homeostasis. (authors)

  14. Altered Th17 cells and Th17/regulatory T-cell ratios indicate the subsequent conversion from undifferentiated connective tissue disease to definitive systemic autoimmune disorders.

    Science.gov (United States)

    Szodoray, Peter; Nakken, Britt; Barath, Sandor; Csipo, Istvan; Nagy, Gabor; El-Hage, Fadi; Osnes, Liv T; Szegedi, Gyula; Bodolay, Edit

    2013-12-01

    A shift in the balance between Th17-cells and regulatory T-cells (Treg) is an important feature of systemic autoimmune diseases (SAID), and may also contribute to their development. Hereby, we assessed the distribution of peripheral Th17 and Treg-cells in patients with undifferentiated connective tissue disease (UCTD), the forerunner of SAIDs and followed these parameters during the development towards definitive SAIDs. Fifty-one UCTD patients were investigated and followed-up for 3 years. Flow cytometry was used to identify and follow three cell-populations: Th17-cells (CD4+IL-17+ T-cells), natural regulatory T-cells (CD4(+)CD25(bright)FoxP3(+); nTregs) and IL-10 producing Type-1 regulatory T-cells (CD4+IL-10+ T-cells; Tr1). Altogether 37.3% of these patients progressed into SAIDs. Th17-cells were increased in UCTD vs. controls, which further increased in those, whom developed SAIDs eventually. The Th17/nTreg ratio gradually increased from controls through UCTD patients, reaching the highest values in SAID-progressed patients. Regarding the Th17/Tr1 ratios, a similar tendency was observed moreover Th17/Tr1 could distinguish between UCTD patients with, or without subsequent SAID progression in a very early UCTD stage. Various immunoserological markers showed association with Th17 and Th17/nTreg at baseline, indicating the consecutive development of a distinct SAID. The derailed Th17/Treg balance may contribute to disease progression therefore could function as a prognostic marker.

  15. Brain-derived neurotrophic factor as an indicator of chemical neurotoxicity: an animal-free CNS cell culture model.

    Science.gov (United States)

    Woehrling, Elizabeth K; Hill, Eric J; Nagel, David; Coleman, Michael D

    2013-12-01

    Recent changes to the legislation on chemicals and cosmetics testing call for a change in the paradigm regarding the current 'whole animal' approach for identifying chemical hazards, including the assessment of potential neurotoxins. Accordingly, since 2004, we have worked on the development of the integrated co-culture of post-mitotic, human-derived neurons and astrocytes (NT2.N/A), for use as an in vitro functional central nervous system (CNS) model. We have used it successfully to investigate indicators of neurotoxicity. For this purpose, we used NT2.N/A cells to examine the effects of acute exposure to a range of test chemicals on the cellular release of brain-derived neurotrophic factor (BDNF). It was demonstrated that the release of this protective neurotrophin into the culture medium (above that of control levels) occurred consistently in response to sub-cytotoxic levels of known neurotoxic, but not non-neurotoxic, chemicals. These increases in BDNF release were quantifiable, statistically significant, and occurred at concentrations below those at which cell death was measureable, which potentially indicates specific neurotoxicity, as opposed to general cytotoxicity. The fact that the BDNF immunoassay is non-invasive, and that NT2.N/A cells retain their functionality for a period of months, may make this system useful for repeated-dose toxicity testing, which is of particular relevance to cosmetics testing without the use of laboratory animals. In addition, the production of NT2.N/A cells without the use of animal products, such as fetal bovine serum, is being explored, to produce a fully-humanised cellular model.

  16. Effects of three algae-lysing bacterial strains on growth of Anabaena flos-aquae%3株溶藻菌对水华鱼腥藻生长的效应

    Institute of Scientific and Technical Information of China (English)

    孙秀敏; 郑培忠; 万旗东; 沈健英

    2012-01-01

    The effects of 3 algae-lysing bacterial strains(AL-l,AL-6 and AL-8)on the growth and photosynthetic pigments of Anabaena flos-aquae were determined by spectrophotometry. The results showed that all the 3 bacterial strains could significantly inhibit the growth of A. flos-aquae and their inhibiting effects had a certain correlation to their concentrations. Also the 3 bacterial strains could influence for A. flos-aquae's pigments to absorb light spectrum. The 3 bacterial strains were somewhat different in algae-lysing mode:AL-l and AL-6 indirectly played an algae-lysing role through secreting heat-stable and heat-labile substances while AL-8 played an algae-lysing role directly and indirectly.%以分光光度法测定3株溶藻菌(AL-1、AL-6和AL-8)对水华鱼腥藻(Anabaena flos-aquae)生长效应和光合色素的影响.结果表明:3株溶藻菌均能显著抑制水华鱼腥藻的生长,对其生长速率、干重的抑制效应明显,并表现出一定的浓度相关性;且3株溶藻菌均能影响水华鱼腥藻的色素吸收光谱.3株溶藻菌溶藻方式略有不同,AL-1和AL-6可以通过分泌热稳定的物质和热不稳定的物质间接起到溶藻效果,而AL-8可同时通过直接和间接方式溶藻.

  17. Upregulation of immunoproteasome subunits in myositis indicates active inflammation with involvement of antigen presenting cells, CD8 T-cells and IFNΓ.

    Directory of Open Access Journals (Sweden)

    Khetam Ghannam

    Full Text Available OBJECTIVE: In idiopathic inflammatory myopathies (IIM infiltration of immune cells into muscle and upregulation of MHC-I expression implies increased antigen presentation and involvement of the proteasome system. To decipher the role of immunoproteasomes in myositis, we investigated individual cell types and muscle tissues and focused on possible immune triggers. METHODS: Expression of constitutive (PSMB5, -6, -7 and corresponding immunoproteasomal subunits (PSMB8, -9, -10 was analyzed by real-time RT-PCR in muscle biopsies and sorted peripheral blood cells of patients with IIM, non-inflammatory myopathies (NIM and healthy donors (HD. Protein analysis in muscle biopsies was performed by western blot. Affymetrix HG-U133 platform derived transcriptome data from biopsies of different muscle diseases and from immune cell types as well as monocyte stimulation experiments were used for validation, coregulation and coexpression analyses. RESULTS: Real-time RT-PCR revealed significantly increased expression of immunoproteasomal subunits (PSMB8/-9/-10 in DC, monocytes and CD8+ T-cells in IIM. In muscle biopsies, the immunosubunits were elevated in IIM compared to NIM and exceeded levels of matched blood samples. Proteins of PSMB8 and -9 were found only in IIM but not NIM muscle biopsies. Reanalysis of 78 myositis and 20 healthy muscle transcriptomes confirmed these results and revealed involvement of the antigen processing and presentation pathway. Comparison with reference profiles of sorted immune cells and healthy muscle confirmed upregulation of PSMB8 and -9 in myositis biopsies beyond infiltration related changes. This upregulation correlated highest with STAT1, IRF1 and IFNγ expression. Elevation of T-cell specific transcripts in active IIM muscles was accompanied by increased expression of DC and monocyte marker genes and thus reflects the cell type specific involvement observed in peripheral blood. CONCLUSIONS: Immunoproteasomes seem to indicate

  18. EVALUATION OF RED CELL INDICES AND RETICULOCYTE MATURITY INDICES INCLUDING RETICULOCYTE HAEMOGLOBIN CONCENTRATION IN IRON DEFICIENCY ANAEMIA IN ADULT FEMALE POPULATION

    Directory of Open Access Journals (Sweden)

    Anitha Sunkara

    2016-12-01

    Full Text Available BACKGROUND Prevalence of iron deficiency anaemia is higher in both developed, underdeveloped countries and in developing countries particularly in toddlers, adolescent girls and women of childbearing age. Diagnosis of iron deficiency is made by biochemical investigations along with the routine haemogram. Automated analysers in the recent years provide many reticulocyte parameters like Low Fluorescence Ratio (LFR, Medium Fluorescence Ratio (MFR and High Fluorescence Ratio (HFR, Reticulocyte Haemoglobin Concentration-haemoglobin (Ret-He, which not only aid in the diagnosis of Iron Deficiency Anaemia (IDA without the necessity of biochemical investigations, but also help in the follow up of these patients for bone marrow response accurately. The aim of the present study is to evaluate the effectiveness of these parameters in the diagnosis of iron deficiency anaemia in the female population in reproductive age group. MATERIALS AND METHODS The present study included peripheral blood samples from fifty eight women aged between 18-45 years from routine workload including 20 normal (Hb >12.0 g/dL with normal serum ferritin and serum iron levels and 38 iron deficiency anaemia samples (Hb <12.0 g/dL and with low serum ferritin and serum iron levels. RESULTS There was significant differences in the means between the subjects from the control group and IDA group in red cell distribution width, RDW CV (% (13.26±1.07 vs. 18.22±3.34 p-value <0.00001 and immature reticulocyte fraction (% (9.87±5.68 vs. 17.89±9.00 p-value=.000646, reticulocyte haemoglobin concentration (pg (33.34±2.92 vs. 22.0±4.13, p-value=<0.00001. Serum iron and serum ferritin showed statistically significant difference between two groups. CONCLUSIONS The reticulocyte parameters including Ret-He provide an important information in suspected cases of iron deficiency. Routine haemogram can thus be more helpful not only on morphological categorisation of anaemia, but also in knowing the

  19. Expression of lux-genes as an indicator of metabolic activity of cells in model ecosystem studies

    Science.gov (United States)

    Boyandin, A. N.; Popova, L. Yu.

    Quick response to different impacts and easy measurement make the luminescent systems of luminous bacteria an object convenient for application in various fields. Cloning of gene luminescence in different organisms is currently used to study both the survival of microbial cells and the effect of different factors on their metabolic activity, including the environment. A primary test-object in estimating bacteriological contamination of water bodies, Escherichia coli, can be conveniently used as an indicator of bactericidal properties of aquatic ecosystems. The application of Escherichia coli Z905/pPHL7 (lux +) as a marker microorganism can facilitate monitoring the microbiological status of closed biocenoses, including systems with higher organisms. The investigation of various parameters of microecosystems (carbon nutrition type, concentrations of inorganic ions and toxic compounds) shows that the recombinant strain E. coli Z905/pPHL7 can be effectively used as a marker.

  20. WE-E-BRE-10: Level of Breast Cancer Stem Cell Correlated with Tumor Radioresistence: An Indication for Individualized Breast Cancer Therapy Adapted to Cancer Stem Cell Fractions

    Energy Technology Data Exchange (ETDEWEB)

    Qi, S; Pajonk, F; McCloskey, S; Low, D; Kupelian, P; Steinberg, M; Sheng, K [UCLA, Los Angeles, CA (United States)

    2014-06-15

    Purposes: The presence of cancer stem cells (CSCs) in a solid tumor could result in poor tumor control probability. The purposes are to study CSC radiosensitivity parameters α and β and their correlation to CSC levels to understand the underlying radioresistance mechanisms and enable individualized treatment design. Methods: Four established breast cancer cell lines (MCF-7, T47D, MDA-MB-231, and SUM159PT) were irradiated in vitro using single radiation doses of 0, 2, 4, 6, 8 or 10 Gy. The fractions of CSCs in each cell lines were determined using cancer stem cell markers. Mammosphere assays were also performed to better estimate the number of CSCs and represent the CSC repopulation in a human solid tumor. The measured cell surviving fractions were fitted using the Linear-quadratic (LQ) model with independent fitting parameters: α-TC, β-TC (TCs), α-CSC, β-CSC (CSCs), and fs (the percentage of CSCs in each sample). Results: The measured fs increased following the irradiation by MCF-7 (0.1%), T47D (0.9%), MDA-MB-231 (1.18%) and SUM159T (2.46%), while decreasing surviving curve slopes were observed, indicating greater radioresistance, in the opposite order. The fitting yielded the radiosensitive parameters for the MCF-7: α-TC=0.1±0.2Gy{sup −1}, β-TC= 0.08 ±0.14Gy{sup −2}, α-CSC=0.04±0.07Gy{sup −1}, β-CSC =0.02±0.3Gy{sup −2}; for the SUM159PT, α-TC=0.08±0.25 Gy{sup −1}, β-TC=0.02±0.02Gy{sup −2}, α-CSC=0.04±0.18Gy{sup −1}, β-CSC =0.004±0.24Gy{sup −2}. In the mammosphere assay, where fs were higher than the corresponding cell line assays, there was almost no shoulder found in the surviving curves (more radioresistant in mammosphere assays) yielding β-CSC of approximately 0. Conclusion: Breast cancer stem cells were more radioresistant characterized by smaller α and β values compared to differentiated breast cancer cells. Percentage of breast cancer stem cells strongly correlated to overall tumor radioresistance. This observation

  1. Characterization of Thermophilic Strain SY-14 with Capability to Lyse Bacterial Cells%嗜热溶胞菌SY-14的基本特性研究

    Institute of Scientific and Technical Information of China (English)

    宋玉栋; 胡洪营; 席劲瑛

    2007-01-01

    对从污泥堆肥中分离得到的具有溶胞能力的嗜热产芽孢杆菌SY-14进行了初步鉴定,并对其生长、产酶及溶胞特性进行了研究.根据菌株形态及16S rDNA测序结果,SY-14为土芽胞杆菌属(Geobacillus sp.).SY-14最适生长温度60℃左右,最适生长pH为6.0~7.0.SY-14培养上清液对完整和加热灭活后的微生物细胞都具有溶胞能力,6 h的细胞溶解率分别可达到70%和85%,上清液经过热处理后溶胞能力明显下降,表明溶胞能力主要来自于酶的作用.间歇培养过程中,SY-14培养上清液溶胞活性在对数生长期快速升高,进入稳定期前后达到最大,之后快速下降.SY-14培养上清液对受试的5株革兰氏阴性菌及部分革兰氏阳性菌都具有溶胞能力.

  2. A chimeric LysK-lysostaphin fusion enzyme lysing Staphylococcus aureus cells: a study of both kinetics of inactivation and specifics of interaction with anionic polymers

    Science.gov (United States)

    A staphylolytic fusion protein (K-L) was created, harboring three unique lytic activities comprised of the LysK CHAP endopeptidase, and amidase domains, and the lysostaphin glycyl-glycine endopeptidase domain. To assess the potential of possible therapeutic applications, the kinetic behavior of K-L...

  3. Correlation of discocyte frequency and ATP concentration in preserved blood: A morphological indicator of red blood cell viability

    Directory of Open Access Journals (Sweden)

    Leonart M.S.S.

    1997-01-01

    Full Text Available Red blood cells (RBC are viable if kept in an adequate preservative solution, although gradual changes in morphology and metabolism may occur. There is a gradual decrease in adenosine-5'-triphosphate (ATP concentration, pH, glucose consumption, and enzyme activity during preservation. The normal discocyte shapes are initially replaced by echinocytes and stomatocytes and, at final stages, by spherocytes, the last step before splenic sequestration. Post-transfusional survival has been correlated with the ATP concentration. RBC preserved in ADSOL, a solution containing adenine, dextrose, sodium chloride, and mannitol, are viable for transfusion for up to 6 weeks. Erythrocytes from 10 blood units taken from healthy adult donors were preserved for 12 weeks in ADSOL at 4oC. We now report a significant correlation (r2 = 0.98 between the percentage of discocytes (89 to 7% and ATP (100 to 10% concentration in ADSOL-preserved RBC. The results suggest that the percent of discocyte shapes used as an indicator of ATP concentration may be a useful indicator for quality control of RBC viability in centers which have limited assay facilities.

  4. Drug-induced death of leukaemic cells after G2/M arrest: higher order DNA fragmentation as an indicator of mechanism.

    OpenAIRE

    1998-01-01

    Many reports have documented apoptotic death in different cell types within hours of exposure to cytotoxic drugs; lower drug concentrations may cause cell cycle arrest at G2/M and subsequent death, which has been distinguished from 'classic' apoptosis. We have analysed etoposide-induced cell death in two lymphoblastoid T-cell lines, CCRF-CEM and MOLT-4, specifically in relation to DNA cleavage as indicated by pulse-field gel and conventional electrophoresis. High (5 microM) concentration etop...

  5. Analysis of Gene Expression Profiles of Microdissected Cell Populations Indicates that Testicular Carcinoma In situ Is an Arrested Gonocyte

    DEFF Research Database (Denmark)

    Sonne, S. B.; Almstrup, K.; Dalgaard, M.

    2009-01-01

    performed laser microdissection of CIS cells. Highly enriched cell populations were obtained and subjected to gene expression analysis. The expression profile of CIS cells was compared with microdissected gonocytes, oogonia, and cultured embryonic stem cells with and without genomic aberrations. Three...... a meiotic cell, and the similarity to embryonic stem cells was modest compared with gonocytes. Thus, we provide new evidence that the molecular phenotype of CIS cells is similar to that of gonocytes. Our data are in line with the idea that CIS cells may be gonocytes that survived in the postnatal testis....... speculate that disturbed development of somatic cells in the fetal testis may play a role in allowing undifferentiated cells to survive in the postnatal testes. The further development of CIS into invasive germ cell tumors may depend on signals from their postpubertal niche of somatic cells, including...

  6. Cell wall ultrastructure of flocculent and non-flocculent Schizosaccharomyces pombe strains. Effect of cell wall hydrolysing enzymes on flocculation and cell wall ultastructure.

    Science.gov (United States)

    Geleta, Anna; Kristóf, Z; Maráz, Anna

    2007-03-01

    Scanning and transmission electron microscopic studies revealed the presence of slime-like, amorphous material on the surface of Schizosaccahromyces pombe RIVE 4-2-1 cells, independently, whether they were in flocculated or in non-flocculated state. Close contact of the adjacent cells via the merging outermost cell wall layers was found, however, only in the case of floc formation, which was induced by cultivating the cells in the presence of 6% (v/v) ethanol. Irreversible loss of the flocculation ability of the cells by treatment with proteinases suggests that proteinaceous cell surface molecules as lectins contribute to the cell-to-cell interaction during flocculation. Both proteinase K and pronase treatments removed a distinct outer layer of the cell wall, which indicated that the protein moieties of the phosphogalactomannan outer surface layer has a crucial role in the maintenance of cell wall integrity. In the case of lysing enzyme treatment the removal of the outermost layer was also observed as the first step of the cell wall digestion, while driselase treatment resulted in almost complete digestion of the cell wall.

  7. A quality monitoring program for red blood cell components: in vitro quality indicators before and after implementation of semiautomated processing.

    Science.gov (United States)

    Acker, Jason P; Hansen, Adele L; Kurach, Jayme D R; Turner, Tracey R; Croteau, Ioana; Jenkins, Craig

    2014-10-01

    Canadian Blood Services has been conducting quality monitoring of red blood cell (RBC) components since 2005, a period spanning the implementation of semiautomated component production. The aim was to compare the quality of RBC components produced before and after this production method change. Data from 572 RBC units were analyzed, categorized by production method: Method 1, RBC units produced by manual production methods; Method 2, RBC units produced by semiautomated production and the buffy coat method; and Method 3, RBC units produced by semiautomated production and the whole blood filtration method. RBC units were assessed using an extensive panel of in vitro tests, encompassing regulated quality control criteria such as hematocrit (Hct), hemolysis, and hemoglobin (Hb) levels, as well as adenosine triphosphate, 2,3-diphosphoglycerate, extracellular K(+) and Na(+) levels, methemoglobin, p50, RBC indices, and morphology. Throughout the study, all RBC units met mandated Canadian Standards Association guidelines for Hb and Hct, and most (>99%) met hemolysis requirements. However, there were significant differences among RBC units produced using different methods. Hb content was significantly lower in RBC units produced by Method 2 (51.5 ± 5.6 g/unit; p < 0.001). At expiry, hemolysis was lowest in Method 2-produced RBC units (p < 0.05) and extracellular K(+) levels were lowest in units produced by Method 1 (p < 0.001). While overall quality was similar before and after the production method change, the observed differences, although small, indicate a lack of equivalency across RBC products manufactured by different methods. © 2014 AABB.

  8. A Practical Optimisation Method to Improve QOS and GOS-Based Key Performance Indicators in GSM Network Cell Cluster Environment [

    Directory of Open Access Journals (Sweden)

    Joseph Isabona

    2014-11-01

    Full Text Available The delivering of both good quality of service (QoS and Grade of Service (GoS in any competitive mob ile communication environment is a major factor to redu cing subscribers’ churn rate. Therefore, it is important for wireless mobile network operators to ensure stability and efficiency by delivering a consistent, reliable and high-quality end user (sub scriber satisfaction. This can only be achieve by conducting a regular network performance monitoring and optimisation as it directly impacts the qualit y of the offered services and hence user satisfaction. I n this paper, we present the results of network performance evaluation and optimisation of a GSM ne twork on cell cluster-basis, in Asaba region, South East Nigeria. We employ a combination of essential key performance indicators such as dropped call rat e, call setup success rate and outage call rate to exa mine overall QoS and GoS performance of the GSM network. Our results after network optimisation sho wed significant performance improvement in terms of call drop rate, call set up success rate, and call block rate across. Specifically, the end user satis faction rate has increased from 94.45%, 87.74%, and 92.85% to 99.05%, 95.38% and 99.03% respectively across the three GSM cell clusters. The GoS is reduced fro m 3.33%, 6.60% and 2.38% to 0.00%, 3.70% and 0.00% respectively. Furthermore, ESA, which corresp ond end points service availability, has improved from 94.44%, 93.40% and 97.62% to 100%, 96.30% and 100% respectively. In addition, the average throughput has improved from 73.74kbits/s, 85.06kbi ts/s and 87.54kbits/s to 77.07kbits/s, 92.38kbits/s and 102kbits/s respectively across the three GSM cell c lusters.

  9. Microbial community analysis of natural water and preliminary study on algae-lysing protease%城市自然水体微生物种群分析及溶藻蛋白酶初步研究

    Institute of Scientific and Technical Information of China (English)

    吴雅丽; 傅以钢

    2013-01-01

    The PCR-DGGE was used to analyze the microbial community structure in natural waters and the influence on the microbial community structure changes by impact of the different concentrations of Microcystic aeruginosa. At the same time, the function of using the algae to induce the algae-lysing protease in the microorganisms was investigated. The re?sults showed that the diversity of microorganisms in natural waters was identified and the microbial community structure did not changed obviously under the impact of algae, but the succession of some dominant bacteria was observed. The algae-lysing protease was induced by the dosing algae and the protease in the Clostridium botulinum was basically determined as algae-lysing protease by sequence analysis of the protein fragment.%采用PCR-DGGE技术,研究了城市自然水体中微生物菌群结构及投加不同浓度的铜绿微囊藻对水体微生物种群结构变化的影响,同时探讨了藻类诱导水体中微生物产生溶藻蛋白酶的作用;结果表明自然水体中微生物多样性十分丰富,加藻冲击后水体微生物种群结构没有明显变化,个别菌种的优势发生演替;微生物溶藻蛋白酶被诱导,通过片段序列分析,可以基本确定为微生物Clostridium botulinum体内溶藻酶.

  10. HIF1-alpha overexpression indicates a good prognosis in early stage squamous cell carcinomas of the oral floor

    Directory of Open Access Journals (Sweden)

    Joos Ulrich

    2005-07-01

    Full Text Available Abstract Background Hypoxia-inducible factor 1 (HIF-1 is a transcription factor, which plays a central role in biologic processes under hypoxic conditions, especially concerning tumour angiogenesis. HIF-1α is the relevant, oxygen-dependent subunit and its overexpression has been associated with a poor prognosis in a variety of malignant tumours. Therefore, HIF-1α expression in early stage oral carcinomas was evaluated in relation to established clinico-pathological features in order to determine its value as a prognostic marker. Methods 85 patients with histologically proven surgically treated T1/2 squamous cell carcinoma (SCC of the oral floor were eligible for the study. Tumor specimens were investigated by means of tissue micro arrays (TMAs and immunohistochemistry for the expression of HIF-1. Correlations between clinical features and the expression of HIF-1 were evaluated by Kaplan-Meier curves, log-rank tests and multivariate Cox regression analysis. Results HIF-1α was frequently overexpressed in a probably non-hypoxia related fashion. The expression of HIF-1α was related with a significantly improved 5-year survival rate (p Conclusion HIF-1α overexpression is an indicator of favourable prognosis in T1 and T2 SCC of the oral floor. Node negative patients lacking HIF-1α expression may therefore be considered for adjuvant radiotherapy.

  11. A spatial hazard model for cluster detection on continuous indicators of disease: application to somatic cell score.

    Science.gov (United States)

    Gay, Emilie; Senoussi, Rachid; Barnouin, Jacques

    2007-01-01

    Methods for spatial cluster detection dealing with diseases quantified by continuous variables are few, whereas several diseases are better approached by continuous indicators. For example, subclinical mastitis of the dairy cow is evaluated using a continuous marker of udder inflammation, the somatic cell score (SCS). Consequently, this study proposed to analyze spatialized risk and cluster components of herd SCS through a new method based on a spatial hazard model. The dataset included annual SCS for 34 142 French dairy herds for the year 2000, and important SCS risk factors: mean parity, percentage of winter and spring calvings, and herd size. The model allowed the simultaneous estimation of the effects of known risk factors and of potential spatial clusters on SCS, and the mapping of the estimated clusters and their range. Mean parity and winter and spring calvings were significantly associated with subclinical mastitis risk. The model with the presence of 3 clusters was highly significant, and the 3 clusters were attractive, i.e. closeness to cluster center increased the occurrence of high SCS. The three localizations were the following: close to the city of Troyes in the northeast of France; around the city of Limoges in the center-west; and in the southwest close to the city of Tarbes. The semi-parametric method based on spatial hazard modeling applies to continuous variables, and takes account of both risk factors and potential heterogeneity of the background population. This tool allows a quantitative detection but assumes a spatially specified form for clusters.

  12. Destruction of solid tumors by immune cells

    Science.gov (United States)

    López, Álvaro G.; Seoane, Jesús M.; Sanjuán, Miguel A. F.

    2017-03-01

    The fractional cell kill is a mathematical expression describing the rate at which a certain population of cells is reduced to a fraction of itself. In order to investigate the fractional cell kill that governs the rate at which a solid tumor is lysed by a cell population of cytotoxic CD8+ T cells (CTLs), we present several in silico simulations and mathematical analyses. When the CTLs eradicate efficiently the tumor cells, the models predict a correlation between the morphology of the tumors and the rate at which they are lysed. However, when the effectiveness of the immune cells is decreased, the mathematical function fails to reproduce the process of lysis. This limit is thoroughly discussed and a new fractional cell kill is proposed.

  13. [Application of the Ca2+ indicator fluo-3 and fluo-4 in the process of H2O2 induced apoptosis of A549 cell].

    Science.gov (United States)

    Zhang, Siyang; Li, Chunyan; Gao, Jian; Qiu, Xueshan; Cui, Zeshi

    2014-03-01

    Lung cancer is a common malignant tumor all over the world, and Ca(2+) is a critical regulator for apoptosis of cancer cells. The monitoring of cytoplastic Ca(2+) level in real-time will contribute to further investigate the molecular mechanisms of apoptosis mediated by Ca2+ in lung cancer cells. To evaluate the Ca(2+) indicator fluo-3 and fluo-4 in the process of H2O2 induced the apoptosis of lung adenocarcinoma A549 cells. The cytoplastic Ca(2+) concentration ([Ca(2+)]i) was determined in real-time, and the correlations between [Ca(2+)]i and cell apoptosis were investigated. The differences in fluorescence intensity and measured value were compared between the two Ca(2+) indicators. Cells were loaded with the Ca(2+) indicator fluo-3 or fluo-4 for 1 h, and then stimulated with 50 mM H2O2. Laser scanning confocal microscope was applied to perform real-time monitoring on the variation of [Ca(2+)]i in selected cells. DAPI staining was used to observe apoptosis in H2O2 treated cells. Our results showed that the fluorescence intensity of fluo-4 was stronger than that of fluo-3 in the same condition of dye concentration, loading time and image acquisition parameters before or after H2O2 stimulation. The cytoplastic [Ca(2+)]i was rapidly elevated in H2O2 stimulated A549 cells. The range of [Ca(2+)]i in selected cells loaded with fluo-3 was 112.2 nM-1,069.6 nM, and that in selected cells loaded with fluo-4 was 7.6 nM-505.4 nM. Moreover, the apoptotic rate was significantly increased in H2O2 treated cells, compared with untreated ones (Pfluo-4 was probably more applicable to measure [Ca(2+)]i in cells with less content of Ca(2+).

  14. Design of a high-throughput human neural crest cell migration assay to indicate potential developmental toxicants.

    Science.gov (United States)

    Nyffeler, Johanna; Karreman, Christiaan; Leisner, Heidrun; Kim, Yong Jun; Lee, Gabsang; Waldmann, Tanja; Leist, Marcel

    2017-01-01

    Migration of neural crest cells (NCCs) is one of the pivotal processes of human fetal development. Malformations arise if NCC migration and differentiation are impaired genetically or by toxicants. In the currently available test systems for migration inhibition of NCC (MINC), the manual generation of a cell-free space results in extreme operator dependencies, and limits throughput. Here a new test format was established. The assay avoids scratching by plating cells around a commercially available circular stopper. Removal of the stopper barrier after cell attachment initiates migration. This microwell-based circular migration zone NCC function assay (cMINC) was further optimized for toxicological testing of human pluripotent stem cell (hPSC)-derived NCCs. The challenge of obtaining data on viability and migration by automated image processing was addressed by developing a freeware. Data on cell proliferation were obtained by labelling replicating cells, and by careful assessment of cell viability for each experimental sample. The role of cell proliferation as an experimental confounder was tested experimentally by performing the cMINC in the presence of the proliferation-inhibiting drug cytosine arabinoside (AraC), and by a careful evaluation of mitotic events over time. Data from these studies led to an adaptation of the test protocol, so that toxicant exposure was limited to 24 h. Under these conditions, a prediction model was developed that allows classification of toxicants as either inactive, leading to unspecific cytotoxicity, or specifically inhibiting NC migration at non-cytotoxic concentrations.

  15. Fluorescence-based co-culture of normal and cancerous cells as an indicator of therapeutic effects in cancer.

    Science.gov (United States)

    Tamura, Masato; Matsui, Hirofumi; Hyodo, Ichinosuke; Tanaka, Junko; Miwa, Yoshihiro

    2014-10-15

    Comprehensive evaluation of the effects of cancer therapies in vitro is difficult because of the need to distinguish the main effects from the side effects within the data. This problem cannot be overcome by methods involving monoculture, because the effects of anti-cancer drugs in a monoculture can only be measured on either normal or cancerous cells in isolation. In order to promote therapeutic development, therefore, we need a novel drug evaluation method which can simultaneously determine both therapeutic activity and toxicity under a co-culture of normal and cancerous cells. Co-culture creates a more biomimetic condition in comparison to monoculture. The novel method proposed in this study uses an easy experiment for estimating the effects of treatments with various kinds of drugs as a solution to the abovementioned problems. We have previously established two cell lines: a rat gastric mucosal cell line (RGM) and its corresponding cancerous mutant cell line (RGK). In this study, we have developed a new evaluation procedure using a co-culture of green fluorescent protein-expressing RGM cells (RGM-GFP) and kusabira orange-expressing RGK cells (RGK-KO). These cell lines emit green and red fluorescence, respectively. We demonstrated the capability of the method in evaluations of the cancer-selective effects of anti-cancer drugs and X-ray treatment. These results clearly distinguished the cancer-selective toxicity of the applied therapies.

  16. DNA damage induced by cis- and carboplatin as indicator for in vitro sensitivity of ovarian carcinoma cells

    Directory of Open Access Journals (Sweden)

    de Wilde Rudy L

    2009-10-01

    Full Text Available Abstract Background The DNA damage by platinum cytostatics is thought to be the main cause of their cytotoxicity. Therefore the measurement of the DNA damage induced by cis- and carboplatin should reflect the sensitivity of cancer cells toward the platinum chemotherapeutics. Methods DNA damage induced by cis- and carboplatin in primary cells of ovarian carcinomas was determined by the alkaline comet assay. In parallel, the reduction of cell viability was measured by the fluorescein diacetate (FDA hydrolysis assay. Results While in the comet assay the isolated cells showed a high degree of DNA damage after a 24 h treatment, cell viability revealed no cytotoxicity after that incubation time. The individual sensitivities to DNA damage of 12 tumour biopsies differed up to a factor of about 3. DNA damage after a one day treatment with cis- or carboplatin correlated well with the cytotoxic effects after a 7 day treatment (r = 0,942 for cisplatin r = 0.971 for carboplatin. In contrast to the platinum compounds the correlation of DNA damage and cytotoxicity induced by adriamycin was low (r = 0,692, or did not exist for gemcitabine. Conclusion The measurement of DNA damage induced by cis- and carboplatin is an accurate method to determine the in vitro chemosensitivity of ovarian cancer cells towards these cytostatics, because of its quickness, sensitivity, and low cell number needed.

  17. Application of the Ca2+ Indicator Fluo-3 and Fluo-4 in the Process of H2O2 
Induced Apoptosis of A549 Cell

    Directory of Open Access Journals (Sweden)

    Siyang ZHANG

    2014-03-01

    Full Text Available Background and objective Lung cancer is a common malignant tumor all over the world, and Ca2+ is a critical regulator for apoptosis of cancer cells. The monitoring of cytoplastic Ca2+ level in real-time will contribute to further investigate the molecular mechanisms of apoptosis mediated by Ca2+ in lung cancer cells. To evaluate the Ca2+ indicator fluo-3 and fluo-4 in the process of H2O2 induced the apoptosis of lung adenocarcinoma A549 cells. The cytoplastic Ca2+ concentration ([Ca2+]i was determined in real-time, and the correlations between [Ca2+]i and cell apoptosis were investigated. The differences in fluorescence intensity and measured value were compared between the two Ca2+ indicators. Methods Cells were loaded with the Ca2+ indicator fluo-3 or fluo-4 for 1 h, and then stimulated with 50 mM H2O2. Laser scanning confocal microscope was applied to perform real-time monitoring on the variation of [Ca2+]i in selected cells. DAPI staining was used to observe apoptosis in H2O2 treated cells. Results Our results showed that the fluorescence intensity of fluo-4 was stronger than that of fluo-3 in the same condition of dye concentration, loading time and image acquisition parameters before or after H2O2 stimulation. The cytoplastic [Ca2+]i was rapidly elevated in H2O2 stimulated A549 cells. The range of [Ca2+]i in selected cells loaded with fluo-3 was 112.2 nM-1,069.6 nM, and that in selected cells loaded with fluo-4 was 7.6 nM-505.4 nM. Moreover, the apoptotic rate was significantly increased in H2O2 treated cells, compared with untreated ones (P<0.01. Conclusion In summary, H2O2 promoted Ca2+ release in A549 cells, and induced cell apoptosis. Ca2+ indicator fluo-4 was probably more applicable to measure [Ca2+]i in cells with less content of Ca2+.

  18. Evaluation of the removal of indicator bacteria from domestic sludge processed by Autothermal Thermophilic Aerobic Digestion (ATAD).

    Science.gov (United States)

    Piterina, Anna V; Bartlett, John; Pembroke, Tony J

    2010-09-01

    The degradation of sludge solids in an insulated reactor during Autothermal Thermophilic Aerobic Digestion (ATAD) processing results in auto-heating, thermal treatment and total solids reduction, however, the ability to eliminate pathogenic organisms has not been analysed under large scale process conditions. We evaluated the ATAD process over a period of one year in a two stage, full scale Irish ATAD plant established in Killarney and treating mixed primary and secondary sludge, by examining the sludge microbiologically at various stages during and following ATAD processing to determine its ability to eliminate indicator organisms. Salmonella spp. (pathogen) and fecal-coliform (indicator) densities were well below the limits used to validate class A biosolids in the final product. Enteric pathogens present at inlet were deactivated during the ATAD process and were not detected in the final product using both traditional microbial culture and molecular phylogenetic techniques. A high DNase activity was detected in the bulk sludge during the thermophilic digestion stage which may be responsible for the rapid turn over of DNA from lysed cells and the removal of mobile DNA. These results offer assurance for the safe use of ATAD sludge as a soil supplement following processing.

  19. Evaluation of the Removal of Indicator Bacteria from Domestic Sludge Processed by Autothermal Thermophilic Aerobic Digestion (ATAD

    Directory of Open Access Journals (Sweden)

    Tony J. Pembroke

    2010-09-01

    Full Text Available The degradation of sludge solids in an insulated reactor during Autothermal Thermophilic Aerobic Digestion (ATAD processing results in auto-heating, thermal treatment and total solids reduction, however, the ability to eliminate pathogenic organisms has not been analysed under large scale process conditions. We evaluated the ATAD process over a period of one year in a two stage, full scale Irish ATAD plant established in Killarney and treating mixed primary and secondary sludge, by examining the sludge microbiologically at various stages during and following ATAD processing to determine its ability to eliminate indicator organisms. Salmonella spp. (pathogen and fecal-coliform (indicator densities were well below the limits used to validate class A biosolids in the final product. Enteric pathogens present at inlet were deactivated during the ATAD process and were not detected in the final product using both traditional microbial culture and molecular phylogenetic techniques. A high DNase activity was detected in the bulk sludge during the thermophilic digestion stage which may be responsible for the rapid turn over of DNA from lysed cells and the removal of mobile DNA. These results offer assurance for the safe use of ATAD sludge as a soil supplement following processing.

  20. Folate-conjugated immunoglobulin targets melanoma tumor cells for NK cell effector functions

    Science.gov (United States)

    Skinner, Cassandra C.; McMichael, Elizabeth L.; Jaime-Ramirez, Alena C.; Abrams, Zachary B.; Lee, Robert J.; Carson, William E.

    2016-01-01

    The folate receptor (FR) is over-expressed on the vascular side of cancerous cells including those of the breast, ovaries, testes, and cervix. We hypothesized that a folate-conjugated immunoglobulin (F-IgG) would bind to the FR that is over-expressed on melanoma tumor cells to target these cells for lysis by natural killer (NK) cells. Folate receptor expression was confirmed in the Mel-39 (human melanoma) cell line by flow cytometry and immunoblot analysis, using KB (human oral epithelial) and F01 (human melanoma) as a positive and negative control, respectively. FR-positive and negative cell lines were treated with F-IgG or control immunoglobulin G (C-IgG) in the presence or absence of cytokines in order to determine NK cell ability to lyse FR-positive cell lines. NK cell activation was significantly upregulated and lysis of Mel 39 tumor cells enhanced following treatment with F-IgG, as compared to C-IgG at all effector:target (E:T) ratios (p<0.01). This trend was further enhanced by NK cell stimulation with the activating cytokine interleukin-12 (IL-12). NK cell production of cytokines such as interferon-gamma (IFN-γ), macrophage inflammatory protein 1 alpha (MIP-1α), and regulated on activation normal T-cell expressed and secreted (RANTES) were also significantly increased in response to co-stimulation with IL-12 stimulation and F-IgG-coated Mel 39 target cells, as compared to controls (p<0.01). In contrast, F-IgG did not bind to the FR-negative cell line F01 and had no significant effect on NK cell lysis or cytokine production. This research indicates the potential use of F-IgG for its ability to induce an immune response from NK cells against FR-positive melanoma tumor cells which can be further enhanced by the addition of cytokines. PMID:27035691

  1. Waste indicators

    Energy Technology Data Exchange (ETDEWEB)

    Dall, O.; Lassen, C.; Hansen, E. [Cowi A/S, Lyngby (Denmark)

    2003-07-01

    The Waste Indicator Project focuses on methods to evaluate the efficiency of waste management. The project proposes the use of three indicators for resource consumption, primary energy and landfill requirements, based on the life-cycle principles applied in the EDIP Project. Trial runs are made With the indicators on paper, glass packaging and aluminium, and two models are identified for mapping the Danish waste management, of which the least extensive focuses on real and potential savings. (au)

  2. Quality indicators

    DEFF Research Database (Denmark)

    Hjorth-Andersen, Christian

    1991-01-01

    In recent literature it has been suggested that consumers need have no knowledge of product quality as a number of quality indicators (or signals) may be used as substitutes. Very little attention has been paid to the empirical verification of these studies. The present paper is devoted...... to the issue of how well these indicators perform, using market data provided by consumer magazines from 3 countries. The results strongly indicate that price is a poor quality indicator. The paper also presents some evidence which suggests that seller reputation and easily observable characteristics are also...

  3. Programmed cell death as a prognostic indicator for radiation therapy in cervical carcinoma patients: A pilot study

    Directory of Open Access Journals (Sweden)

    Bhosle S

    2005-01-01

    Full Text Available Purpose: In clinical practice, radiation therapy often fails in cervical carcinoma stage IIIB and there is a need to develop a predictive assay for prognosis of radiation treatment outcome in cancer patient. We have attempted to evaluate the relevance of changes in Membrane Fluidity (MF and associated apoptotic cell death in cervical cancer cells after first fractionated dose of radiation therapy to treatment outcome of stage IIIB cervical carcinoma patients. Materials and Methods: Biopsies of 15 patients with histologically proven cervix cancer were collected from the patients before and 24 h after first fractionated radiation dose of 2 grays (Gy. Cell suspension made in Dulbecco′s Modified Eagle′s Medium (DMEM were used for further investigations and cell suspension of cervix cancer patient were used to measure MF by fluorescence polarization method and apoptotic index (AI was determined by Tdt dUTP Nucleotide End Labeling (TUNEL assay. Results: A substantial increase in MF and AI was observed in cervical cancer cells irradiated ex vivo . A significant correlation ( P < 0.001 was found between the changes in AI after first fractionated dose of radiotherapy and treatment outcome of patients. No significant correlation ( P > 0.1 was detected between changes in MF and treatment outcome of patients. Conclusion: Preliminary results showed significant change in MF and a marked increase in percentage apoptosis of cervix cancer cells irradiated ex vivo . The changes in AI after first fractionated dose of radiotherapy in cervical carcinoma patients may provide a predictor of prognosis for radiotherapy in uterine cervical carcinoma patients.

  4. MART-10 represses cholangiocarcinoma cell growth and high vitamin D receptor expression indicates better prognosis for cholangiocarcinoma

    Science.gov (United States)

    Chiang, Kun-Chun; Yeh, Ta-Sen; Huang, Cheng-Cheng; Chang, Yu-Chan; Juang, Horng-Heng; Cheng, Chi-Tung; Pang, Jong-Hwei S.; Hsu, Jun-Te; Takano, Masashi; Chen, Tai C.; Kittaka, Atsushi; Hsiao, Michael; Yeh, Chun-Nan

    2017-01-01

    Cholangiocarcinoma (CCA) is a devastating disease due to no effective treatments available. Since the non-mineral functions of vitamin D emerges, 1α,25(OH)2D3, the active form of vitamin D, has been applied in anti-cancer researches. In this study, we demonstrated that both the 1α,25(OH)2D3 analog, MART-10, and 1α,25(OH)2D3 possessed anti-growth effect on human CCA cells with MART-10 much more potent than 1α,25(OH)2D3. The growth inhibition of both drugs were mediated by induction of G0/G1 cell cycle arrest through upregulation of p27 and downregulation of CDK4, CDK6, and cyclin D3. Human neutrophil gelatinase associated lipocalin (NGAL) was found to be involved in 1α,25(OH)2D3 and MART-10 meditated growth inhibition for CCA as knockdown of NGAL decreased Ki-67 expression in SNU308 cells and rendered SNU308 cells less responsive to 1α,25(OH)2D3 and MART-10 treatment. Vitamin D receptor (VDR) knockdown partly abolished MART-10-induced inhibition of NGAL and cell growth in SNU308 cells. The xenograft animal study demonstrated MART-10 could effectively repressed CCA growth in vivo without inducing obvious side effects. The IHC examination of human CCA specimen for VDR revealed that higher VDR expression was linked with better prognosis. Collectively, our results suggest that MART-10 could be a promising regimen for CCA treatment. PMID:28256614

  5. Retention of Ag-specific memory CD4(+) T cells in the draining lymph node indicates lymphoid tissue resident memory populations.

    Science.gov (United States)

    Marriott, Clare L; Dutton, Emma E; Tomura, Michio; Withers, David R

    2017-03-15

    Several different memory T-cell populations have now been described based upon surface receptor expression and migratory capabilities. Here we have assessed murine endogenous memory CD4(+) T cells generated within a draining lymph node and their subsequent migration to other secondary lymphoid tissues. Having established a model response targeting a specific peripheral lymph node, we temporally labelled all the cells within draining lymph node using photoconversion. Tracking of photoconverted and non-photoconverted Ag-specific CD4(+) T cells revealed the rapid establishment of a circulating memory population in all lymph nodes within days of immunisation. Strikingly, a resident memory CD4(+) T cell population became established in the draining lymph node and persisted for several months in the absence of detectable migration to other lymphoid tissue. These cells most closely resembled effector memory T cells, usually associated with circulation through non-lymphoid tissue, but here, these cells were retained in the draining lymph node. These data indicate that lymphoid tissue resident memory CD4(+) T-cell populations are generated in peripheral lymph nodes following immunisation.

  6. Investigation of influences of the paraformaldehyde fixation and paraffin embedding removal process on refractive indices and scattering properties of epithelial cells

    Science.gov (United States)

    Su, Jing-Wei; Hsu, Wei-Chen; Tjiu, Jeng-Wei; Chiang, Chun-Pin; Huang, Chao-Wei; Sung, Kung-Bin

    2014-07-01

    The scattering properties and refractive indices (RI) of tissue are important parameters in tissue optics. These parameters can be determined from quantitative phase images of thin slices of tissue blocks. However, the changes in RI and structure of cells due to fixation and paraffin embedding might result in inaccuracies in the estimation of the scattering properties of tissue. In this study, three-dimensional RI distributions of cells were measured using digital holographic microtomography to obtain total scattering cross sections (TSCS) of the cells based on the first-order Born approximation. We investigated the slight loss of dry mass and drastic shrinkage of cells due to paraformaldehyde fixation and paraffin embedding removal processes. We propose a method to compensate for the correlated changes in volume and RI of cells. The results demonstrate that the TSCS of live cells can be estimated using restored cells. The percentage deviation of the TSCS between restored cells and live cells was only -8%. Spatially resolved RI and scattering coefficients of unprocessed oral epithelium ranged from 1.35 to 1.39 and from 100 to 450 cm-1, respectively, estimated from paraffin-embedded oral epithelial tissue after restoration of RI and volume.

  7. Short hairpin RNA screen indicates that Klotho beta/FGF19 protein overcomes stasis in human colonic epithelial cells.

    Science.gov (United States)

    Kim, Jinyong; Eskiocak, Ugur; Stadler, Guido; Lou, Zhenjun; Kuro-o, Makoto; Shay, Jerry W; Wright, Woodring E

    2011-12-16

    Normal human colonic epithelial cells (HCECs) are not immortalized by telomerase alone but also require CDK4. Some human cell types growth-arrest due to stress- or aberrant signaling-induced senescence (stasis). Stasis represents the consequences of growth conditions culture that are inadequate to maintain long-term proliferation. Overexpressed CDK4 titers out p16 and allows cells to ignore the growth arrest signals produced by stasis. To identify factors contributing to the inadequate culture environment, we used a 62,000-member shRNA library to knock down factors cooperating with human telomerase reverse transcriptase (hTERT) in the immortalization of HCECs. Knockdown of Klotho gamma (KLG; also known as KLPH and LCTL) allowed hTERT to immortalize HCECs. KLG is one isoform of the Klotho family of factors that coordinate interaction between different FGF ligands and the FGF receptor. We also found that knockdown of KLG induced another member of the Klotho family, Klotho beta (KLB). Induction of KLB was maintained and could activate ERK1/2 in immortalized cells. Supplementation of the culture medium with the KLB ligand FGF19 had a similar effect on hTERT-expressing HCECs as knockdown of KLG regarding both immortalization and down-regulation of the tumor suppressor Klotho alpha. Together, these data suggest that KLB is an important regulator in the immortalization of HCECs by facilitating FGF19 growth factor signaling.

  8. The H4IIE cell bioassay as an indicator of dioxin-like chemicals in wildlife and the environment.

    Science.gov (United States)

    Whyte, J J; Schmitt, C J; Tillitt, D E

    2004-01-01

    The H4IIE cell bioassay has proven utility as a screening tool for planar halogenated hydrocarbons (PHHs) and structurally similar chemicals accumulated in organisms from the wild. This bioassay has additional applications in hazard assessment of PHH exposed populations. In this review, the toxicological principles, current protocols, performance criteria, and field applications for the assay are described. The H4IIE cell bioassay has several advantages over the analytical measurement of PHHs in environmental samples, but conclusions from studies can be strengthened when both bioassay and analytical chemistry data are presented together. Often, the bioassay results concur with biological effects in organisms and support direct measures of PHHs. For biomonitoring purposes and prioritization of PHH-contaminated environments, the H4IIE bioassay may be faster and less expensive than analytical measurements. The H4IIE cell bioassay can be used in combination with other biomarkers such as in vivo measurements of CYP1A1 induction to help pinpoint the sources and identities of dioxin-like chemicals. The number of studies that measure H4IIE-derived TCDD-EQs continues to increase, resulting in subtle improvements over time. Further experiments are required to determine if TCDD-EQs derived from mammalian cells are adequate predictors of toxicity to non-mammalian species. The H4IIE cell bioassay has been used in over 300 published studies, and its combination of speed, simplicity, and ability to integrate the effects of complex contaminant mixtures makes it a valuable addition to hazard assessment and biomonitoring studies.

  9. Rejection of normal and neoplastic hemopoietic cells by lethally irradiated mice

    Energy Technology Data Exchange (ETDEWEB)

    Afifi, M.S.H.

    1985-01-01

    The objective of this study was to investigate the mechanisms of rejection of normal and neoplastic hemopoietic cells by lethally irradiated mice, in part by investigating the hypothesis that two or more cell types are involved in recognition and rejection of hemopoietic cells. Interferon (IFN) was used as a tool for investigating such mechanisms. IFN alpha/beta stimulated the rejection of normal hemopoietic marrow cell grafts in Fl hybrid and in allogeneic host mice but did not affect the growth of cells in syngeneic mice. IFN alpha/beta was effective in hosts pretreated with silica but not in hosts pretreated with cyclophosphamide (Cy) or with anti-asialoGMI serum. Rabbit anti-IFN alpha/beta, but not anti-IFN gamma, serum inhibited genetic resistance to bone marrow cells. These results indicated that IFN alpha/beta was acting indirectly during the rejection of normal hemopoietic cells. It is proposed that four events occur in succession: a host cell recognizes the hemopoietic histocompatibility (Hh) antigens expressed on the surface of incompatible stem cells; this recognition leads to secretion of IFN; IFN activates natural killer (NK) cells; NK cells lyse donor stem cells. Silica interrupts one or both of the first two events. i.e., recognition and/or interrupts one or both of the first two events, i.e. recognition and/or IGN secretion.

  10. Evaluation of clinical trial eligibility and prognostic indices in a population-based cohort of systemic peripheral T-cell lymphomas from the Danish Lymphoma Registry

    DEFF Research Database (Denmark)

    Pedersen, Martin Bjerregaard; Hamilton-Dutoit, Stephen Jacques; Bendix, Knud;

    2015-01-01

    patients, approximately half were eligible for multiagent chemotherapy with or without consolidating SCT. Both IPI and PIT are useful prognostic indices in all 'primary nodal' PTCL entities. The prognostic value of ALK protein expression in anaplastic large cell lymphoma is significantly downsized when...

  11. Solar Indices

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Collection includes a variety of indices related to solar activity contributed by a number of national and private solar observatories located worldwide. This...

  12. Analysis of COPII Vesicles Indicates a Role for the Emp47-Ssp120 Complex in Transport of Cell Surface Glycoproteins.

    Science.gov (United States)

    Margulis, Neil G; Wilson, Joshua D; Bentivoglio, Christine M; Dhungel, Nripesh; Gitler, Aaron D; Barlowe, Charles

    2016-03-01

    Coat protein complex II (COPII) vesicle formation at the endoplasmic reticulum (ER) transports nascent secretory proteins forward to the Golgi complex. To further define the machinery that packages secretory cargo and targets vesicles to Golgi membranes, we performed a comprehensive proteomic analysis of purified COPII vesicles. In addition to previously known proteins, we identified new vesicle proteins including Coy1, Sly41 and Ssp120, which were efficiently packaged into COPII vesicles for trafficking between the ER and Golgi compartments. Further characterization of the putative calcium-binding Ssp120 protein revealed a tight association with Emp47 and in emp47Δ cells Ssp120 was mislocalized and secreted. Genetic analyses demonstrated that EMP47 and SSP120 display identical synthetic positive interactions with IRE1 and synthetic negative interactions with genes involved in cell wall assembly. Our findings support a model in which the Emp47-Ssp120 complex functions in transport of plasma membrane glycoproteins through the early secretory pathway.

  13. Analysis of Papaya Cell Wall-Related Genes during Fruit Ripening Indicates a Central Role of Polygalacturonases during Pulp Softening

    Science.gov (United States)

    Fabi, João Paulo; Broetto, Sabrina Garcia; da Silva, Sarah Lígia Garcia Leme; Zhong, Silin; Lajolo, Franco Maria; do Nascimento, João Roberto Oliveira

    2014-01-01

    Papaya (Carica papaya L.) is a climacteric fleshy fruit that undergoes dramatic changes during ripening, most noticeably a severe pulp softening. However, little is known regarding the genetics of the cell wall metabolism in papayas. The present work describes the identification and characterization of genes related to pulp softening. We used gene expression profiling to analyze the correlations and co-expression networks of cell wall-related genes, and the results suggest that papaya pulp softening is accomplished by the interactions of multiple glycoside hydrolases. The polygalacturonase cpPG1 appeared to play a central role in the network and was further studied. The transient expression of cpPG1 in papaya results in pulp softening and leaf necrosis in the absence of ethylene action and confirms its role in papaya fruit ripening. PMID:25162506

  14. Stochastic modeling indicates that aging and somatic evolution in the hematopoetic system are driven by non-cell-autonomous processes.

    Science.gov (United States)

    Rozhok, Andrii I; Salstrom, Jennifer L; DeGregori, James

    2014-12-01

    Age-dependent tissue decline and increased cancer incidence are widely accepted to be rate-limited by the accumulation of somatic mutations over time. Current models of carcinogenesis are dominated by the assumption that oncogenic mutations have defined advantageous fitness effects on recipient stem and progenitor cells, promoting and rate-limiting somatic evolution. However, this assumption is markedly discrepant with evolutionary theory, whereby fitness is a dynamic property of a phenotype imposed upon and widely modulated by environment. We computationally modeled dynamic microenvironment-dependent fitness alterations in hematopoietic stem cells (HSC) within the Sprengel-Liebig system known to govern evolution at the population level. Our model for the first time integrates real data on age-dependent dynamics of HSC division rates, pool size, and accumulation of genetic changes and demonstrates that somatic evolution is not rate-limited by the occurrence of mutations, but instead results from aged microenvironment-driven alterations in the selective/fitness value of previously accumulated genetic changes. Our results are also consistent with evolutionary models of aging and thus oppose both somatic mutation-centric paradigms of carcinogenesis and tissue functional decline. In total, we demonstrate that aging directly promotes HSC fitness decline and somatic evolution via non-cell-autonomous mechanisms.

  15. Protection of cells against oxidative stress by nanomolar levels of hydroxyflavones indicates a new type of intracellular antioxidant mechanism.

    Directory of Open Access Journals (Sweden)

    Emanuele Lombardo

    Full Text Available Natural polyphenol compounds are often good antioxidants, but they also cause damage to cells through more or less specific interactions with proteins. To distinguish antioxidant activity from cytotoxic effects we have tested four structurally related hydroxyflavones (baicalein, mosloflavone, negletein, and 5,6-dihydroxyflavone at very low and physiologically relevant levels, using two different cell lines, L-6 myoblasts and THP-1 monocytes. Measurements using intracellular fluorescent probes and electron paramagnetic resonance spectroscopy in combination with cytotoxicity assays showed strong antioxidant activities for baicalein and 5,6-dihydroxyflavone at picomolar concentrations, while 10 nM partially protected monocytes against the strong oxidative stress induced by 200 µM cumene hydroperoxide. Wide range dose-dependence curves were introduced to characterize and distinguish the mechanism and targets of different flavone antioxidants, and identify cytotoxic effects which only became detectable at micromolar concentrations. Analysis of these dose-dependence curves made it possible to exclude a protein-mediated antioxidant response, as well as a mechanism based on the simple stoichiometric scavenging of radicals. The results demonstrate that these flavones do not act on the same radicals as the flavonol quercetin. Considering the normal concentrations of all the endogenous antioxidants in cells, the addition of picomolar or nanomolar levels of these flavones should not be expected to produce any detectable increase in the total cellular antioxidant capacity. The significant intracellular antioxidant activity observed with 1 pM baicalein means that it must be scavenging radicals that for some reason are not eliminated by the endogenous antioxidants. The strong antioxidant effects found suggest these flavones, as well as quercetin and similar polyphenolic antioxidants, at physiologically relevant concentrations act as redox mediators to enable

  16. Homeobox gene expression profile indicates HOXA5 as a candidate prognostic marker in oral squamous cell carcinoma.

    Science.gov (United States)

    Rodini, Camila Oliveira; Xavier, Flávia Caló Aquino; Paiva, Katiúcia Batista Silva; De Souza Setúbal Destro, Maria Fernanda; Moyses, Raquel Ajub; Michaluarte, Pedro; Carvalho, Marcos Brasilino; Fukuyama, Erica Erina; Tajara, Eloiza Helena; Okamoto, Oswaldo Keith; Nunes, Fabio Daumas

    2012-04-01

    The search for molecular markers to improve diagnosis, individualize treatment and predict behavior of tumors has been the focus of several studies. This study aimed to analyze homeobox gene expression profile in oral squamous cell carcinoma (OSCC) as well as to investigate whether some of these genes are relevant molecular markers of prognosis and/or tumor aggressiveness. Homeobox gene expression levels were assessed by microarrays and qRT-PCR in OSCC tissues and adjacent non-cancerous matched tissues (margin), as well as in OSCC cell lines. Analysis of microarray data revealed the expression of 147 homeobox genes, including one set of six at least 2-fold up-regulated, and another set of 34 at least 2-fold down-regulated homeobox genes in OSCC. After qRT-PCR assays, the three most up-regulated homeobox genes (HOXA5, HOXD10 and HOXD11) revealed higher and statistically significant expression levels in OSCC samples when compared to margins. Patients presenting lower expression of HOXA5 had poorer prognosis compared to those with higher expression (P=0.03). Additionally, the status of HOXA5, HOXD10 and HOXD11 expression levels in OSCC cell lines also showed a significant up-regulation when compared to normal oral keratinocytes. Results confirm the presence of three significantly upregulated (>4-fold) homeobox genes (HOXA5, HOXD10 and HOXD11) in OSCC that may play a significant role in the pathogenesis of these tumors. Moreover, since lower levels of HOXA5 predict poor prognosis, this gene may be a novel candidate for development of therapeutic strategies in OSCC.

  17. Red blood cell count as an indicator of microvascular complications in Chinese patients with type 2 diabetes mellitus

    Directory of Open Access Journals (Sweden)

    Wang ZS

    2013-05-01

    Full Text Available Zhan-Sheng Wang,1,2 Zhan-Chun Song,1 Jing-Hui Bai,1 Fei Li,3 Tao Wu,1 Ji Qi,2 Jian Hu11Department of Cardiology, First Affiliated Hospital of China Medical University, 2Second Department of Cardiology, Fourth People's Hospital of Shenyang, Shenyang, 3Department of Cardiology, Shenzhou Hospital of Shenyang Medical College, Shenyang, People's Republic of ChinaBackground: Rheological disorders of red blood cells (RBC and decreased RBC deformability have been involved in the development of diabetic microangiopathy. However, few studies have evaluated the association of RBC count with microvascular complications in patients with type 2 diabetes mellitus (T2DM. The purpose of this study was to investigate the association of RBC count with microvascular complications in patients with T2DM.Methods: This study involved 369 patients with T2DM: 243 with one or more microvascular complications and 126 without microvascular complications. Anticoagulated blood was collected and analyzed in an automated blood cell counter. The presence of risk factors for microvascular complications was determined.Results: The proportion of patients with microvascular complications increased as the RBC count decreased (P < 0.001. After adjustment for known risk factors for microvascular complications by logistic regression analysis, lower quartiles of RBC count were associated with a higher risk of microvascular complications compared with the reference group composed of the highest quartile (first quartile, odds ratio 4.98, 95% confidence interval 1.54–6.19, P = 0.008; second quartile, odds ratio 3.21, 95% confidence interval 1.17–5.28, P = 0.024.Conclusion: A decreased RBC count is associated with microvascular complications in Chinese patients with T2DM. The RBC count is a potential marker to improve further the ability to identify diabetic patients at high risk of microvascular complications.Keywords: red blood cell count, microvascular complication, type 2 diabetes

  18. The identification of lymphocyte-like cells and lymphoid-related genes in amphioxus indicates the twilight for the emergence of adaptive immune system.

    Directory of Open Access Journals (Sweden)

    Gonghua Huang

    Full Text Available To seek evidence of a primitive adaptive immune system (AIS before vertebrate, we examined whether lymphocytes or lymphocyte-like cells and the related molecules participating in the lymphocyte function existed in amphioxus. Anatomical analysis by electron microscopy revealed the presence of lymphocyte-like cells in gills, and these cells underwent morphological changes in response to microbial pathogens that are reminiscent of those of mammalian lymphocytes executing immune response to microbial challenge. In addition, a systematic comparative analysis of our cDNA database of amphioxus identified a large number of genes whose vertebrate counterparts are involved in lymphocyte function. Among these genes, several genes were found to be expressed in the vicinity of the lymphocyte-like cells by in situ hybridization and up-regulated after exposure to microbial pathogens. Our findings in the amphioxus indicate the twilight for the emergence of AIS before the invertebrate-vertebrate transition during evolution.

  19. A brief report of basic science: the effects of preincisional low-dose ketamine on natural killer cell activity in male Fischer 344 rats after intra-abdominal surgery.

    Science.gov (United States)

    Estes, Savannah; Dinh, Tim; Garrett, Normalynn

    2009-01-01

    Although the first line of defense in cancer treatment often is surgery, studies suggest that postoperative pain and anesthetic drugs suppress the activity of cells that lyse metastatic cells, that is, natural killer cells. We assessed the affect of low-dose ketamine on natural killer cell activity. The findings are presented in this brief report.

  20. Cell-to-cell spread and massive vacuole formation after Cryptococcus neoformans infection of murine macrophages

    OpenAIRE

    Casadevall Arturo; Alvarez Mauricio

    2007-01-01

    Abstract Background The interaction between macrophages and Cryptococcus neoformans (Cn) is critical for containing dissemination of this pathogenic yeast. However, Cn can either lyse macrophages or escape from within them through a process known as phagosomal extrusion. Both events result in live extracellular yeasts capable of reproducing and disseminating in the extracellular milieu. Another method of exiting the intracellular confines of cells is through host cell-to-cell transfer of the ...

  1. High-throughput microfluidic device for single cell analysis using multiple integrated soft lithographic pumps.

    Science.gov (United States)

    Patabadige, Damith E W; Mickleburgh, Tom; Ferris, Lorin; Brummer, Gage; Culbertson, Anne H; Culbertson, Christopher T

    2016-05-01

    The ability to accurately control fluid transport in microfluidic devices is key for developing high-throughput methods for single cell analysis. Making small, reproducible changes to flow rates, however, to optimize lysis and injection using pumps external to the microfluidic device are challenging and time-consuming. To improve the throughput and increase the number of cells analyzed, we have integrated previously reported micropumps into a microfluidic device that can increase the cell analysis rate to ∼1000 cells/h and operate for over an hour continuously. In order to increase the flow rates sufficiently to handle cells at a higher throughput, three sets of pumps were multiplexed. These pumps are simple, low-cost, durable, easy to fabricate, and biocompatible. They provide precise control of the flow rate up to 9.2 nL/s. These devices were used to automatically transport, lyse, and electrophoretically separate T-Lymphocyte cells loaded with Oregon green and 6-carboxyfluorescein. Peak overlap statistics predicted the number of fully resolved single-cell electropherograms seen. In addition, there was no change in the average fluorescent dye peak areas indicating that the cells remained intact and the dyes did not leak out of the cells over the 1 h analysis time. The cell lysate peak area distribution followed that expected of an asynchronous steady-state population of immortalized cells.

  2. Multifocal Langerhans cell histiocytosis of bone: Indications for radiotherapy; Histiocytose langerhansienne osseuse multifocale: place de la radiotherapie

    Energy Technology Data Exchange (ETDEWEB)

    Gaundong Mbethe, G.L.; Dejean, C.; Henriques de Figueiredo, B.; Sargos, P.; Italiano, A.; Kantor, G. [Service de radiotherapie, centre regional de lutte contre le cancer, institut Bergonie, 33 - Bordeaux (France)

    2010-12-15

    Langerhans cell histiocytosis is a non-malignant proliferative disease of unknown etiology that can affect one or more organs. This is a rare disease, 1 to 2/100, 000, affecting mainly children with a male predominance. The osseous lesions are the most frequent (60 to 90%). There is however no consensus treatment for the management of these sites. We report the cases of two patients successfully treated with radiotherapy after primary chemotherapy, at doses of 15 Gy in ten sessions of 1.5 Gy for one patient and 18 Gy in ten fractions of 1.8 Gy for the other. Single or multifocal bone Langerhans cell histiocytosis without visceral involvement is a benign, self-limiting affection in most cases. Some bone lesions could be treated by radiotherapy alone. But the high variability of doses currently given in the literature does not allow determining the lowest effective dose limiting the risk of secondary neoplasia or impaired growth in children, in whom lower doses of 6 to 8 Gy are recommended. The decision of radiotherapy must be weighed against the risk of the disease. Caution should be the rule in this non-malignant tumour pathology. (authors)

  3. Analysis of microRNA expression in canine mammary cancer stem-like cells indicates epigenetic regulation of transforming growth factor-beta signaling.

    Science.gov (United States)

    Rybicka, A; Mucha, J; Majchrzak, K; Taciak, B; Hellmen, E; Motyl, T; Krol, M

    2015-02-01

    Cancer stem cells (CSCs) display both unique self-renewal ability as well as the ability to differentiate into many kinds of cancer cells. They are supposed to be responsible for cancer initiation, recurrence and drug resistance. Despite the fact that a variety of methods are currently employed in order to target CSCs, little is known about the regulation of their phenotype and biology by miRNAs. The aim of our study was to assess miRNA expression in canine mammary cancer stem-like cells (expressing stem cell antigen 1, Sca-1; CD44 and EpCAM) sorted from canine mammary tumour cell lines (CMT-U27, CMT-309 and P114). In order to prove their stem-like phenotype, we conducted a colony formation assay that confirmed their ability to form colonies from a single cell. Profiles of miRNA expression were investigated using Agilent custom-designed microarrays. The results were further validated by real-time rt-PCR analysis of expression of randomly selected miRNAs. Target genes were indicated and analysed using Kioto Encyclopedia of Genes and Genomes (KEGG) and BioCarta databases. The results revealed 24 down-regulated and nine up-regulated miRNAs in cancer stem-like cells compared to differentiated tumour cells. According to KEGG and BioCarta databases, target genes (n=240) of significantly down-regulated miRNAs were involved in transforming growth factor-beta signaling, mitogen-activated protein kinases (MAPK) signaling pathway, anaplastic lymphoma receptor tyrosine kinase (ALK) and peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PGC1A) pathways. The analysis of single-gene overlapping with different pathways showed that the most important genes were: TGFBR1, TGFBR2, SOS1, CHUK, PDGFRA, SMAD2, MEF2A, MEF2C and MEF2D. All of them are involved in tumor necrosis factor-beta signaling and may indicate its important role in cancer stem cell biology. Increased expression of TGFBR2, SMAD2, MEF2A and MEF2D in canine mammary cancer stem-like cells was further

  4. Fasting blood glucose is a novel prognostic indicator for extranodal natural killer/T-cell lymphoma, nasal type

    Science.gov (United States)

    Cai, Q; Luo, X; Liang, Y; Rao, H; Fang, X; Jiang, W; Lin, T; Lin, T; Huang, H

    2013-01-01

    Background: Extranodal natural killer (NK)/T-cell lymphoma, nasal type (ENKTL) is an aggressive disease with poor prognosis, requiring risk stratification. However, the prognosis of ENKTL is not fully defined and needs supplementation. We hypothesised that fasting blood glucose (FBG) may be a new prognostic factor for ENKTL. Methods: We retrospectively analysed 130 patients newly diagnosed with ENKTL. Results: Both univariate analysis and multivariate analysis revealed that FBG >100 mg dl−1 was associated with a poor outcome. Patients with FBG >100 mg dl−1 at diagnosis had more adverse clinical features, achieved lower complete remission rates (P=0.003) and had worse overall survival (PKPI) scores with different survival outcomes (PKPI. PMID:23299534

  5. Comparative effects of the blue green algae Nodularia spumigena and a lysed extract on detoxification and antioxidant enzymes in the green lipped mussel (Perna viridis)

    Energy Technology Data Exchange (ETDEWEB)

    Davies, Warren R. [Department of Biotechnology and Environmental Biology, RMIT University, Melbourne, Victoria (Australia)]. E-mail: warren.davies@rmit.edu.au; Siu, William H.L. [Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong (China); Jack, Ralph W. [Department of Microbiology, University of Otago, Dunedin (New Zealand); Wu, Rudolf S.S. [Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong (China); Lam, Paul K.S. [Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong (China); Nugegoda, Dayanthi [Department of Biotechnology and Environmental Biology, RMIT University, Melbourne, Victoria (Australia)

    2005-07-01

    Nodularia spumigena periodically proliferates to cause toxic algal blooms with some aquatic animals enduring and consuming high densities of the blue green algae or toxic lysis. N. spumigena contains toxic compounds such as nodularin and lipopolysaccharides. This current work investigates physiological effects of exposure from bloom conditions of N. spumigena cells and a post-bloom lysis. Biochemical and antioxidative biomarkers were comparatively studied over an acute 3-day exposure. In general, a post-bloom N. spumigena lysis caused opposite physiological responses to bloom densities of N. spumigena. Specifically, increases in glutathione (GSH) and glutathione peroxidase (GPx) and decreases in glutathione S-transferase (GST) were observed from the N. spumigena lysis. In contrast, N. spumigena cell densities decreased GSH and increased GST and lipid peroxidation (LPO) in mussels. Findings also suggest that at different stages of a toxic bloom, exposure may result in toxic stress to specific organs in the mussel.

  6. Attenuation fluctuations and local dermal reflectivity are indicators of immune cell infiltrate and epidermal hyperplasia in skin inflammation

    Science.gov (United States)

    Phillips, Kevin G.; Wang, Yun; Choudhury, Niloy; Levitz, David; Swanzey, Emily; Lagowski, James; Kulesz-Martin, Molly; Jacques, Steven

    2012-02-01

    Psoriasis is a common inflammatory skin disease resulting from genetic and environmental alterations of cutaneous immune responses responsible for skin homeostasis. While numerous therapeutic targets involved in the immunopathogenesis of psoriasis have been identified, the in vivo dynamics of psoriasis remains under investigated. To elucidate the spatial-temporal morphological evolution of psoriasis we undertook in vivo time course focus-tracked optical coherence tomography (OCT) imaging to non-invasively document dermal alterations due to immune cell infiltration and epidermal hyperplasia in an Imiquimod (IMQ) induced model of psoriasis-like inflammation in DBA2/C57Bl6 hybrid mice. Quantitative appraisal of dermal architectural changes was achieved through a three parameter fit of OCT axial scans in the dermis of the form A(z) = ρ exp(-mu;z +ɛ(z)). Ensemble averaging of the fit parameters over 2000 axial scans per mouse in each treatment arm revealed that the local dermal reflectivity ρ, decreased significantly in response to 6 day IMQ treatment (p = 0.0001), as did the standard deviation of the attenuation fluctuation std(ɛ(z)), (p = 0.04), in comparison to cream controls and day 1 treatments. No significant changes were observed in the average dermal attenuation rate, μ. Our results suggest these label-free OCT-based metrics can be deployed to investigate new therapeutic targets in animal models as well as aid in clinical staging of psoriasis in conjunction with the psoriasis area and severity index.

  7. Integrative proteomics and tissue microarray profiling indicate the association between overexpressed serum proteins and non-small cell lung cancer.

    Directory of Open Access Journals (Sweden)

    Yansheng Liu

    Full Text Available Lung cancer is the leading cause of cancer deaths worldwide. Clinically, the treatment of non-small cell lung cancer (NSCLC can be improved by the early detection and risk screening among population. To meet this need, here we describe the application of extensive peptide level fractionation coupled with label free quantitative proteomics for the discovery of potential serum biomarkers for lung cancer, and the usage of Tissue microarray analysis (TMA and Multiple reaction monitoring (MRM assays for the following up validations in the verification phase. Using these state-of-art, currently available clinical proteomic approaches, in the discovery phase we confidently identified 647 serum proteins, and 101 proteins showed a statistically significant association with NSCLC in our 18 discovery samples. This serum proteomic dataset allowed us to discern the differential patterns and abnormal biological processes in the lung cancer blood. Of these proteins, Alpha-1B-glycoprotein (A1BG and Leucine-rich alpha-2-glycoprotein (LRG1, two plasma glycoproteins with previously unknown function were selected as examples for which TMA and MRM verification were performed in a large sample set consisting about 100 patients. We revealed that A1BG and LRG1 were overexpressed in both the blood level and tumor sections, which can be referred to separate lung cancer patients from healthy cases.

  8. Flow cytometry analysis of FITC-labeled concanavalin A binding to human blood cells as an indicator of radiation-induced membrane alterations

    Energy Technology Data Exchange (ETDEWEB)

    Donnadieu-Claraz, M.; Paillole, N.; Voisin, P. [CEA Centre d`Etudes de Fontenay-aux-Roses, 92 (France). Dept. de Protection de la Sante de l`Homme et de Dosimetrie; Djounova, J. [National Centre of Radiobiology and Radiation Protection, Sofia (Bulgaria)

    1995-12-31

    The {sup 3}H concanavalin-A binding to human blood cells have been described as a promising biological indicator of radiation overexposure. Flow cytometry adaptation of this technique using fluorescein-labelled concanavalin-A were performed to estimate time-dependent changes in binding on human blood cells membranes after in vitro {gamma} irradiation ({sup 60}Co). Result revealed significant enhanced lectin-binding to platelets and erythrocytes in a dose range of 0,5-5 Gy, 1 and 3 hours after irradiation. However for both platelets and erythrocytes, it was impossible to discriminate between the different doses. Further studies are necessary to confirm the suitability of lectin-binding as a biological indicator for radiation dose assessment. (authors). 5 refs., 1 fig.

  9. Studies on nonidet P40 lysis of murine lymphoid cells. I. Use of cholera toxin and cell surface Ig to determine degree of dissociation of the plasma membrane.

    Science.gov (United States)

    Hart, D A

    1975-09-01

    Lymphoid cells from A/J mice were iodinated (125I) by the lactoperoxidase lysed with the non-ionic detergent NP-40. The plasma membrane glycolipid receptor for cholera toxin and cell surface immunoglobulin were utilized in immune precipitation systems to characterize the degree of dissociation of the plasma membrane under various conditions. It was found that at 0.1% NP-40 and at cell concentration from 5 to 10 times 10(7) cells/ml, lipid-protein and protein-lipid-protein complexes formed in NP-40 which were soluble after centrifugation at 10(5) times G. Column chromatography of 125I-cell lysates on agarose A-0.5 M in 0.1% or 0.5% NP-40/PBS indicated that the majority of iodinated cell surface material existed as aggregates in detergent micelles. The availability of the oligosaccharide moiety of the glycolipid to interact with the cholera toxin was dependent on both the detergent concentration and the cell concentration used for cell lysis. However, the cell surface immunoglobulin was immunoprecipitable under all conditions of lysis tested.

  10. Tattoo Pigments Are Observed in the Kupffer Cells of the Liver Indicating Blood-Borne Distribution of Tattoo Ink.

    Science.gov (United States)

    Sepehri, Mitra; Sejersen, Tobias; Qvortrup, Klaus; Lerche, Catharina M; Serup, Jørgen

    2017-01-01

    Tattoo pigments are deposited in the skin and known to distribute to regional lymph nodes. Tattoo pigments are small particles and may be hypothesized to reach the blood stream and become distributed to peripheral organs. This has not been studied in the past. The aim of the study was to trace tattoo pigments in internal organs in mice extensively tattooed with 2 different tattoo ink products. Three groups of mice were studied, i.e., 10 tattooed black, 10 tattooed red, and 5 untreated controls. They were tattooed on the entire back with commercial tattoo inks, black and red. Mice were sacrificed after 1 year. Samples were isolated from tattooed skin, lymph nodes, liver, spleen, kidney, and lung. Samples were examined for deposits of tattoo pigments by light microscopy and transmission electron microscopy (TEM). TEM identified intracellular tattoo pigments in the skin and in lymph nodes. TEM in both groups of tattooed mice showed tattoo pigment deposits in the Kupffer cells in the liver, which is a new observation. TEM detected no pigment in other internal organs. Light microscopy showed dense pigment in the skin and in lymph nodes but not in internal organs. The study demonstrated black and red tattoo pigment deposits in the liver; thus, tattoo pigment distributed from the tattooed skin via the blood stream to this important organ of detoxification. The finding adds a new dimension to tattoo pigment distribution in the body, i.e., as observed via the blood in addition to the lymphatic pathway. © 2017 S. Karger AG, Basel.

  11. Co-expression of CD147 and GLUT-1 indicates radiation resistance and poor prognosis in cervical squamous cell carcinoma.

    Science.gov (United States)

    Huang, Xin-Qiong; Chen, Xiang; Xie, Xiao-Xue; Zhou, Qin; Li, Kai; Li, Shan; Shen, Liang-Fang; Su, Juan

    2014-01-01

    The aim of this study was to investigate the association of CD147 and GLUT-1, which play important roles in glycolysis in response to radiotherapy and clinical outcomes in patients with locally advanced cervical squamous cell carcinoma (LACSCC). The records of 132 female patients who received primary radiation therapy to treat LACSCC at FIGO stages IB-IVA were retrospectively reviewed. Forty-seven patients with PFS (progression-free survival) of less than 36 months were regarded as radiation-resistant. Eighty-five patients with PFS longer than 36 months were regarded as radiation-sensitive. Using pretreatment paraffin-embedded tissues, we evaluated CD147 and GLUT-1 expression by immunohistochemistry. Overexpression of CD147, GLUT-1, and CD147 and GLUT-1 combined were 44.7%, 52.9% and 36.5%, respectively, in the radiation-sensitive group, and 91.5%, 89.4% and 83.0%, respectively, in the radiation-resistant group. The 5-year progress free survival (PFS) rates in the CD147-low, CD147-high, GLUT-1-low, GLUT-1-high, CD147- and/or GLUT-1-low and CD147- and GLUT-1- dual high expression groups were 66.79%, 87.10%, 52.78%, 85.82%, 55.94%, 82.90% and 50.82%, respectively. CD147 and GLUT-1 co-expression, FIGO stage and tumor diameter were independent poor prognostic factors for patients with LACSCC in multivariate Cox regression analysis. Patients with high expression of CD147 alone, GLUT-1 alone or co-expression of CD147 and GLUT-1 showed greater resistance to radiotherapy and a shorter PFS than those with low expression. In particular, co-expression of CD147 and GLUT-1 can be considered as a negative independent prognostic factor.

  12. Barefoot Plantar Pressure Indicates Progressive Neurological Damage in Patients with Human T-Cell Lymphotropic Virus Type 1 Infection.

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    Beatriz Helena B Vasconcelos

    Full Text Available The human T-Cell Lymphotropic Virus Type 1 (HTLV-1 is a retrovirus associated with neurological alterations; individuals with HTLV-1 infection may develop HTLV-1 associated myelopathy / tropical spastic paraparesis (HAM/TSP. Frequent neurological complaints include foot numbness and leg weakness. In this study, we compared the distribution of the body weight on different areas of the foot in HTLV-1 patients with HAM/TSP, asymptomatic HTLV-1 patients, and healthy individuals.We studied 36 HTLV-1 infected patients, who were divided in two groups of 18 patients each based on whether or not they had been diagnosed with HAM/TSP, and 17 control subjects. The evaluation included an interview on the patient's clinical history and examinations of the patient's reflexes, foot skin tactile sensitivity, and risk of falling. The pressure distribution on different areas of the foot was measured with baropodometry, using a pressure platform, while the patients had their eyes open or closed.The prevalence of neurological disturbances-altered reflexes and skin tactile sensitivity and increased risk of falling-was higher in HTLV-1 HAM/TSP patients than in HTLV-1 asymptomatic patients. The medium and maximum pressure values were higher in the forefoot than in the midfoot and hindfoot in both HTLV-1 groups. In addition, the pressure on the hindfoot was lower in HAM/TSP patients compared to control subjects.The neurological disturbances associated with HTLV-1 infection gradually worsened from HTLV-1 asymptomatic patients to HAM/TSP patients. Baropodometry is a valuable tool to establish the extent of neurological damage in patients suffering from HTLV-1 infection.

  13. In vivo characterization of genetic expression of virus-transduced calcium indicators in retinal ganglion cells using a low-cost funduscope.

    Science.gov (United States)

    Chang, Yao-Chuan; Walston, Steven T; Chow, Robert H; Weiland, James D

    2016-08-01

    Virus-transduced calcium indicators are effective reporters of neural activity, offering the advantage of cell-specific labeling. To track the level of in vivo expression of genetically encoded calcium indicators (GECIs) in rodent retina, we developed a noninvasive imaging approach based on a custom-modified low-cost and simple fundus system that enabled us to monitor and characterize in vivo bright-field and fluorescence retinal image. The system clearly resolves individual retinal ganglion cells (RGCs) and axons. RGC fluorescence intensity and number of observable fluorescent cells show a consistent rising trend from week 1 to week 3 after viral injection, indicating a uniform increase of GCaMP6f expression. At defined time points, we prepared wholemount retina mounted on a transparent multielectrode array (MEA) and used calcium imaging to identify the optimal time for studying the responsiveness of RGCs to external electrical stimulation. The results show that the fluorescence-endoscopy fundus system is a powerful and widely accessible tool for evaluating in vivo fluorescence reporter expression.

  14. A confocal study of mechanism of 5-hydroxytryptaminoinduced intracellular calcium dynamics in cultured ratstomach fundus smooth muscle cells with a new Ca2+indicator STDIn-AM

    Institute of Scientific and Technical Information of China (English)

    ZHANG; Xiaoling; (张小玲); YAN; Hongtao; (阎宏涛)

    2002-01-01

    A new fluorescent Ca2+ indicator STDIn-AM for detecting i transients in cultured smooth muscle cells is presented. By making a comparison, the difference between STDIn and fluo-3 is discussed in detail. Using the new Ca2+ indicator, the mechanism of 5-hydroxytryptamino (5-HT) induced intracellular calcium dynamics in stomach fundus smooth muscle cells (SFSMC) of rats is investigated. It is shown that in contrast with fluo-3, STDIn is uniformly distributed in the cytosolic compartment but excluded from the nucleus, when it is transfected into cells. This feature makes it a real cytosol Ca2+ indicator and can reflect changes of cytosol more accurately than that of fluo-3. In addition, STDIn responds to the i transients more sensitive and faster than fluo-3. The results also show that, the L-type Ca2+ channel inhibitor Mn9202 and the PLC inhibitor Compound 48/80 can significantly inhibit the i elevation induced by 5-HT, while the PKC inhibitor D-Sphingosine can enhance the effect of 5-HT. The results suggest that 5-HT acts by the way of 5-HT2 receptors on SFSMC, then through 5-HT2 receptors coupled IP3/Ca2+ and GC/PKC double signal transduction pathways to make Ca2+ release from intracellular Ca2+ stores and Ca2+ influx possibly through L-type calcium channels.

  15. Social indicators.

    Science.gov (United States)

    Sheldon, E B; Parke, R

    1975-05-16

    The notions of social indicators and social accounting, expressed by analogy with the national economic accounts, generated excitement in the 1960's, and the interest continues to grow if we may judge from governmental activity and the publication of programmatic and research papers. But the concepts which focused much of the early enthusiasm gave exaggerated promise of policy applications and provided an unproductive basis for research. The essential theoretical prerequisites for developing a system of social accounts-defining the variables and the interrelationships among them-are missing. It is now realized that evaluation research, particularly experimentation, must be relied on for evaluation of government programs. Through the development and analysis of descriptive time series and the modeling of social processes, we will be able to describe the state of the society and its dynamics and thus improve immensely our ability to state problems in a productive fashion, obtain clues as to promising lines of endeavor, and ask good questions. But these activities cannot measure program effectiveness. Finally, we must be skeptical about definitions of the social indicators enterprise which confine it to social engineering efforts. The issue is not whether social indicators are useful for policy but, rather, how this usefulness comes about. The interest in social indicators has stimulated a revival of interest in quantitative, comparative, social analysis (60), in the analysis of social change, in conceptual and measurement work on such topics as prejudice, crime, and learning, and in the development of models of social processes. The fruit of these efforts will be more directly a contribution to the policy-maker's cognition than to his decisions. Decision emerges from a mosaic of inputs, including valuational and political, as well as technical components. The work we have described deals with only one type of input; it is a contribution to the intellectual mapping

  16. Aberrant plasma IL-7 and soluble IL-7 receptor levels indicate impaired T-cell response to IL-7 in human tuberculosis.

    Directory of Open Access Journals (Sweden)

    Christian Lundtoft

    2017-06-01

    Full Text Available T-cell proliferation and generation of protective memory during chronic infections depend on Interleukin-7 (IL-7 availability and receptivity. Regulation of IL-7 receptor (IL-7R expression and signalling are key for IL-7-modulated T-cell functions. Aberrant expression of soluble (s and membrane-associated (m IL-7R molecules is associated with development of autoimmunity and immune failure in acquired immune deficiency syndrome (AIDS patients. Here we investigated the role of IL-7/IL-7R on T-cell immunity in human tuberculosis. We performed two independent case-control studies comparing tuberculosis patients and healthy contacts. This was combined with follow-up examinations for a subgroup of tuberculosis patients under therapy and recovery. Blood plasma and T cells were characterised for IL-7/sIL-7R and mIL-7R expression, respectively. IL-7-dependent T-cell functions were determined by analysing STAT5 phosphorylation, antigen-specific cytokine release and by analysing markers of T-cell exhaustion and inflammation. Tuberculosis patients had lower soluble IL-7R (p < 0.001 and higher IL-7 (p < 0.001 plasma concentrations as compared to healthy contacts. Both markers were largely independent and aberrant expression normalised during therapy and recovery. Furthermore, tuberculosis patients had lower levels of mIL-7R in T cells caused by post-transcriptional mechanisms. Functional in vitro tests indicated diminished IL-7-induced STAT5 phosphorylation and impaired IL-7-promoted cytokine release of Mycobacterium tuberculosis-specific CD4+ T cells from tuberculosis patients. Finally, we determined T-cell exhaustion markers PD-1 and SOCS3 and detected increased SOCS3 expression during therapy. Only moderate correlation of PD-1 and SOCS3 with IL-7 expression was observed. We conclude that diminished soluble IL-7R and increased IL-7 plasma concentrations, as well as decreased membrane-associated IL-7R expression in T cells, reflect impaired T-cell

  17. Infusion of hemolyzed red blood cells within peripheral blood stem cell grafts in patients with and without sickle cell disease.

    Science.gov (United States)

    Fitzhugh, Courtney D; Unno, Hayato; Hathaway, Vincent; Coles, Wynona A; Link, Mary E; Weitzel, R Patrick; Zhao, Xiongce; Wright, Elizabeth C; Stroncek, David F; Kato, Gregory J; Hsieh, Matthew M; Tisdale, John F

    2012-06-14

    Peripheral blood stem cell (PBSC) infusions are associated with complications such as elevated blood pressure and decreased creatinine clearance. Patients with sickle cell disease experience similar manifestations, and some have postulated release of plasma-free hemoglobin with subsequent nitric oxide consumption as causative. We sought to evaluate whether the infusion of PBSC grafts containing lysed red blood cells (RBCs) leads to the toxicity observed in transplant subjects. We report a prospective cohort study of 60 subjects divided into 4 groups based on whether their infusions contained dimethyl sulfoxide (DMSO) and lysed RBCs, no DMSO and fresh RBCs, DMSO and no RBCs, or saline. Our primary end point, change in maximum blood pressure compared with baseline, was not significantly different among groups. Tricuspid regurgitant velocity and creatinine levels also did not differ significantly among groups. Our data do not support free hemoglobin as a significant contributor to toxicity associated with PBSC infusions. This study was registered at clinicaltrials.gov (NCT00631787).

  18. Unusual expression of red fluorescence at M phase induced by anti-microtubule agents in HeLa cells expressing the fluorescent ubiquitination-based cell cycle indicator (Fucci)

    Energy Technology Data Exchange (ETDEWEB)

    Honda-Uezono, Asumi [Section of Oral Radiation Oncology, Department of Oral Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549 (Japan); Section of Maxillofacial Surgery, Department of Maxillofacial and Neck Reconstruction, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549 (Japan); Kaida, Atsushi [Section of Oral Radiation Oncology, Department of Oral Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549 (Japan); Michi, Yasuyuki; Harada, Kiyoshi [Section of Maxillofacial Surgery, Department of Maxillofacial and Neck Reconstruction, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549 (Japan); Hayashi, Yoshiki; Hayashi, Yoshio [Department of Medicinal Chemistry, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192-0392 (Japan); Miura, Masahiko, E-mail: masa.mdth@tmd.ac.jp [Section of Oral Radiation Oncology, Department of Oral Health Sciences, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549 (Japan)

    2012-11-16

    Highlights: Black-Right-Pointing-Pointer Fucci visualizes cell cycle by green and red fluorescence. Black-Right-Pointing-Pointer Plinabulin, induced unusual red fluorescence at M-phase in HeLa-Fucci cells. Black-Right-Pointing-Pointer The unusual pattern was followed by mitotic catastrophe. Black-Right-Pointing-Pointer The unusual pattern may be an early indicator of cell death in HeLa cells. -- Abstract: Plinabulin (NPI-2358) is a novel microtubule-depolymerizing agent. In HeLa cells, plinabulin arrests the cell-cycle at M phase and subsequently induces mitotic catastrophe. To better understand the effects on this compound on the cell-cycle, we used the fluorescent ubiquitination-based cell cycle indicator (Fucci), which normally enables G1 and S/G2/M cells to emit red and green fluorescence, respectively. When HeLa-Fucci cells were treated with 50 nM plinabulin, cells began to fluoresce both green and red in an unusual pattern; most cells exhibited the new pattern after 24 h of treatment. X-irradiation efficiently induced G2 arrest in plinabulin-treated cells and significantly retarded the emergence of the unusual pattern, suggesting that entering M phase is essential for induction of the pattern. By simultaneously visualizing chromosomes with GFP-histone H2B, we established that the pattern emerges after nuclear envelope breakdown but before metaphase. Pedigree assay revealed a significant relationship between the unusual expression and mitotic catastrophe. Nocodazole, KPU-133 (a more potent derivative of plinabulin), and paclitaxel also exerted similar effects. From these data, we conclude that the unusual pattern may be associated with dysregulation of late M phase-specific E3 ligase activity and mitotic catastrophe following treatment with anti-microtubule agents.

  19. Testicular Cell Indices and Peripheral Blood Testosterone Concentrations in Relation to Age and Semen Quality in Crossbred (Holstein Friesian×Tharparkar Bulls

    Directory of Open Access Journals (Sweden)

    S. K. Rajak

    2014-11-01

    Full Text Available Present study analyzed the changes in peripheral blood testosterone concentrations and testicular cytogram in relation to age and semen quality in crossbred males. Three different age groups of crossbred males viz. bull calves (6 months, n = 5, young bulls (15 months, n = 5 and adult bulls (4 to 6 years, n = 8 were utilized for the study. Testicular fine needle aspiration cytology technique was used to quantify testicular cytology and their indices. Peripheral blood testosterone concentrations were measured using enzyme-linked immunosorbent assay method. Semen samples collected from adult bulls were microscopically evaluated for quality parameters. Mean peripheral blood testosterone concentrations in bull calves, young bulls and adult bulls were 2.28±0.09 ng/mL, 1.42±0.22 ng/mL and 5.66±1.08 ng/mL respectively, and that in adult bulls were significantly different (p<0.01 from young bulls and bull calves. There was no significant difference between the proportion of different testicular cells in bull calves and young bulls. Between young and adult bulls, significant differences (p<0.01 were observed in the proportion of spermatocytes, spermatozoa, and sperm: Sertoli cell ratio. The proportions of Sertoli cells showed a significant difference (p<0.01 between the three age groups. The number of primary spermatocytes had a positive correlation with peripheral blood testosterone concentrations in bull calves (r = 0.719, p<0.01. Number of Sertoli cells per 100 germ cells was negatively correlated with blood testosterone concentration in young bulls (r = −0.713, p<0.01. Among different semen parameters in adult bulls, ejaculate volume (r = 0.790, p<0.05 had positive relationship, and sperm motility had significant negative correlation (r = −0.711, p<0.05 with testosterone concentrations. The number of Sertoli cells and Sertoli cell index had a positive correlation with various semen quality parameters (p<0.001. Results of the present study

  20. Overexpression of Rap-1A indicates a poor prognosis for oral cavity squamous cell carcinoma and promotes tumor cell invasion via Aurora-A modulation.

    Science.gov (United States)

    Chen, Chang-Han; Chuang, Hui-Ching; Huang, Chao-Cheng; Fang, Fu-Min; Huang, Hsuan-Ying; Tsai, Hsin-Ting; Su, Li-Jen; Shiu, Li-Yen; Leu, Steve; Chien, Chih-Yen

    2013-02-01

    The functions of Rap-1A in oral carcinogenesis are largely unexplored. In this study, we examined the expression of Rap-1A at different malignant stages of oral cavity squamous cell carcinoma (OCSCC). Semiquantitative RT-PCR, quantitative RT-PCR, and Western blotting were used to evaluate Rap-1A mRNA and protein expressions, respectively, in paired OCSCC patient specimens. To determine the possible correlation between Rap-1A expression and various clinical characteristics, 256 samples from patients with OCSCC were evaluated by immunohistochemical staining. Strong Rap-1A expression was a significant prognostic marker and predictor of aggressive OCSCC. The overall and disease-specific 5-year survival rates were significantly correlated with strong expression of Rap-1A (P Rap-1A could promote oral cancer cell migration and invasion by Transwell chambers and wound healing assay. Conversely, the suppression of Rap-1A expression using Rap-1A-mediated siRNA was sufficient to decrease cell motility. Furthermore, our data also illustrated that Aurora-A could not only induce mRNA and protein expressions of Rap-1A for enhancing cancer cell motility but also co-localize and form a complex with Rap-1A in the oral cancer cell line. Finally, immunohistochemical staining, indirect immunofluorescence, and Western blotting analysis of human aggressive OCSCC specimens revealed a significantly positive correlation between Rap-1A and Aurora-A expression. Taken together, our results suggest that the Aurora-A/Rap-1A pathway is associated with survival, tumor progression, and metastasis of OCSCC patients. Copyright © 2013 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.

  1. Trichomonas vaginalis and Tritrichomonas foetus: interaction with fibroblasts and muscle cells - new insights into parasite-mediated host cell cytotoxicity

    Directory of Open Access Journals (Sweden)

    Ricardo Chaves Vilela

    2012-09-01

    Full Text Available Trichomonas vaginalis and Tritrichomonas foetus are parasitic, flagellated protists that inhabit the urogenital tract of humans and bovines, respectively. T. vaginalis causes the most prevalent non-viral sexually transmitted disease worldwide and has been associated with an increased risk for human immunodeficiency virus-1 infection in humans. Infections by T. foetus cause significant losses to the beef industry worldwide due to infertility and spontaneous abortion in cows. Several studies have shown a close association between trichomonads and the epithelium of the urogenital tract. However, little is known concerning the interaction of trichomonads with cells from deeper tissues, such as fibroblasts and muscle cells. Published parasite-host cell interaction studies have reported contradictory results regarding the ability of T. foetus and T. vaginalis to interact with and damage cells of different tissues. In this study, parasite-host cell interactions were examined by culturing primary human fibroblasts obtained from abdominal biopsies performed during plastic surgeries with trichomonads. In addition, mouse 3T3 fibroblasts, primary chick embryo myogenic cells and L6 muscle cells were also used as models of target cells. The parasite-host cell cultures were processed for scanning and transmission electron microscopy and were tested for cell viability and cell death. JC-1 staining, which measures mitochondrial membrane potential, was used to determine whether the parasites induced target cell damage. Terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling staining was used as an indicator of chromatin damage. The colorimetric crystal violet assay was performed to ana-lyse the cytotoxicity induced by the parasite. The results showed that T. foetus and T. vaginalis adhered to and were cytotoxic to both fibroblasts and muscle cells, indicating that trichomonas infection of the connective and muscle tissues is likely to occur; such

  2. Mesodermal gene expression in the acoel Isodiametra pulchra indicates a low number of mesodermal cell types and the endomesodermal origin of the gonads.

    Directory of Open Access Journals (Sweden)

    Marta Chiodin

    Full Text Available Acoelomorphs are bilaterally symmetric small marine worms that lack a coelom and possess a digestive system with a single opening. Two alternative phylogenetic positions of this group within the animal tree are currently debated. In one view, Acoelomorpha is the sister group to all remaining Bilateria and as such, is a morphologically simple stepping stone in bilaterian evolution. In the other, the group is a lineage within the Deuterostomia, and therefore, has derived a simple morphology from a more complex ancestor. Acoels and the closely related Nemertodermatida and Xenoturbellida, which together form the Acoelomorpha, possess a very limited number of cell types. To further investigate the diversity and origin of mesodermal cell types we describe the expression pattern of 12 orthologs of bilaterian mesodermal markers including Six1/2, Twist, FoxC, GATA4/5/6, in the acoel Isodiametra pulchra. All the genes are expressed in stem cells (neoblasts, gonads, and at least subsets of the acoel musculature. Most are expressed in endomesodermal compartments of I. pulchra developing embryos similar to what has been described in cnidarians. Our molecular evidence indicates a very limited number of mesodermal cell types and suggests an endomesodermal origin of the gonads and the stem cell system. We discuss our results in light of the two prevailing phylogenetic positions of Acoelomorpha.

  3. Imbalanced protein expression patterns of anabolic, catabolic, anti-catabolic and inflammatory cytokines in degenerative cervical disc cells: new indications for gene therapeutic treatments of cervical disc diseases.

    Directory of Open Access Journals (Sweden)

    Demissew S Mern

    Full Text Available Degenerative disc disease (DDD of the cervical spine is common after middle age and can cause loss of disc height with painful nerve impingement, bone and joint inflammation. Despite the clinical importance of these problems, in current publications the pathology of cervical disc degeneration has been studied merely from a morphologic view point using magnetic resonance imaging (MRI, without addressing the issue of biological treatment approaches. So far a wide range of endogenously expressed bioactive factors in degenerative cervical disc cells has not yet been investigated, despite its importance for gene therapeutic approaches. Although degenerative lumbar disc cells have been targeted by different biological treatment approaches, the quantities of disc cells and the concentrations of gene therapeutic factors used in animal models differ extremely. These indicate lack of experimentally acquired data regarding disc cell proliferation and levels of target proteins. Therefore, we analysed proliferation and endogenous expression levels of anabolic, catabolic, ant-catabolic, inflammatory cytokines and matrix proteins of degenerative cervical disc cells in three-dimensional cultures. Preoperative MRI grading of cervical discs was used, then grade III and IV nucleus pulposus (NP tissues were isolated from 15 patients, operated due to cervical disc herniation. NP cells were cultured for four weeks with low-glucose in collagen I scaffold. Their proliferation rates were analysed using 3-(4, 5-dimethylthiazolyl-2-2,5-diphenyltetrazolium bromide. Their protein expression levels of 28 therapeutic targets were analysed using enzyme-linked immunosorbent assay. During progressive grades of degeneration NP cell proliferation rates were similar. Significantly decreased aggrecan and collagen II expressions (P<0.0001 were accompanied by accumulations of selective catabolic and inflammatory cytokines (disintegrin and metalloproteinase with thrombospondin motifs 4

  4. Differences in indicators of malignancy between luminal epithelial cell type and myoepithelial cell type of simple solid carcinoma in the canine mammary gland.

    Science.gov (United States)

    Yoshimura, H; Nakahira, R; Kishimoto, T E; Michishita, M; Ohkusu-Tsukada, K; Takahashi, K

    2014-11-01

    Routinely diagnosed simple solid carcinoma (SSC) of the canine mammary gland comprises a heterogeneous group of tumors. Seventy-two cases that had been diagnosed as SSC based on hematoxylin and eosin-stained tissue sections were reclassified immunohistochemically on the basis of myoepithelial markers p63 and α-smooth muscle actin, as well as a luminal epithelial marker cytokeratin 8. Only 23 cases (32%) were true SSC, composed only of luminal epithelial cells, whereas 11 cases (15%) were malignant myoepithelioma (MM), composed predominantly of myoepithelial cells, and 38 cases (53%) were biphasic carcinoma (BC), characterized by biphasic proliferation of luminal epithelial and basal/myoepithelial components. As the pathological parameters were compared between the reclassified tumor types, infiltrative potential, vascular/lymphatic invasion, lymph node metastasis, and Ki-67 labeling index were higher in true SSC compared with MM and BC, suggesting that the former may exhibit a poorer prognosis compared with the latter two.

  5. Assay of Histamine in Single Mast Cells by Capillary Zone Electrophoresis with Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Capillary zone electrophoresis was employed for the analysis of histamine in single rat peritoneal mast cells using an amperometric detector. In this method, individual mast cells and then 0.02 mol/L NaOH as a lysing solution are injected into the front end of the separation capillary. A cell injector was constructed for easy injection of single cells. Histamine in single mast cells has been identified and quantified.

  6. Defects in neural stem cell proliferation and olfaction in Chd7 deficient mice indicate a mechanism for hyposmia in human CHARGE syndrome.

    Science.gov (United States)

    Layman, W S; McEwen, D P; Beyer, L A; Lalani, S R; Fernbach, S D; Oh, E; Swaroop, A; Hegg, C C; Raphael, Y; Martens, J R; Martin, D M

    2009-06-01

    Mutations in CHD7, a chromodomain gene, are present in a majority of individuals with CHARGE syndrome, a multiple anomaly disorder characterized by ocular Coloboma, Heart defects, Atresia of the choanae, Retarded growth and development, Genital hypoplasia and Ear anomalies. The clinical features of CHARGE syndrome are highly variable and incompletely penetrant. Olfactory dysfunction is a common feature in CHARGE syndrome and has been potentially linked to primary olfactory bulb defects, but no data confirming this mechanistic link have been reported. On the basis of these observations, we hypothesized that loss of Chd7 disrupts mammalian olfactory tissue development and function. We found severe defects in olfaction in individuals with CHD7 mutations and CHARGE, and loss of odor evoked electro-olfactogram responses in Chd7 deficient mice, suggesting reduced olfaction is due to a dysfunctional olfactory epithelium. Chd7 expression was high in basal olfactory epithelial neural stem cells and down-regulated in mature olfactory sensory neurons. We observed smaller olfactory bulbs, reduced olfactory sensory neurons, and disorganized epithelial ultrastructure in Chd7 mutant mice, despite apparently normal functional cilia and sustentacular cells. Significant reductions in the proliferation of neural stem cells and regeneration of olfactory sensory neurons in the mature Chd7(Gt/+) olfactory epithelium indicate critical roles for Chd7 in regulating neurogenesis. These studies provide evidence that mammalian olfactory dysfunction due to Chd7 haploinsufficiency is linked to primary defects in olfactory neural stem cell proliferation and may influence olfactory bulb development.

  7. Infection with foot-and-mouth disease virus (FMDV) induces a natural killer (NK) cell response in cattle that is lacking following vaccination

    Science.gov (United States)

    Natural killer (NK) cells play a role in innate antiviral immunity by directly lysing virus-infected cells and producing antiviral cytokines such as interferon gamma (IFNgamma). We developed a system for characterizing the bovine NK response to foot-and-mouth disease virus (FMDV), which causes a dis...

  8. NK cell-mediated killing of AML blasts. Role of histamine, monocytes and reactive oxygen metabolites

    Energy Technology Data Exchange (ETDEWEB)

    Brune, M.; Mellqvist, U.H. [Sahlgren`s Univ. Hospital, Dept. of Medicine, Haematology Section, Goeteborg (Sweden); Hansson, M.; Hermodsson, S.; Hellstrand, K. [Sahlgren`s Univ. Hospital, Dept. of Virology, Goeteborg (Sweden)

    1996-10-01

    Blasts recovered from patients with acute myelogenous leukaemia (AML) were lysed by heterologeous natural killer (NK) cells treated with NK cell-activating cytokine-induced killing of AML blasts was inhibited by monocytes, recovered from peripheral blood by counterflow centrifugal elutriation. Histamine, at concentrations exceeding 0.1 {mu}M, abrogated the monocyte-induced inhibition of NK cells; thereby, histamine and IL-2 or histamine and IFN-{alpha} synergistically induced NK cell-mediated destruction of AML blasts. The effect of histamine was completely blocked by the histamine H2-receptor (H2R) antagonist ranitidine but not by its chemical control AH20399AA. Catalase, a scavenger of reactive oxygen metabolites (ROM), reversed the monocyte-induced inhibition of NK cell-mediated killing of blast cells, indicating that the inhibitory signal was mediated by products of the respiratory burst of monocytes. It is concluded that (i) monocytes inhibit anti-leukemic properties of NK cells, (ii) the inhibition is conveyed by monocyte-derived ROM, and (iii) histamine reverses the inhibitory signal and, thereby, synergizes with NK cell-activating cytokines to induce killing of AML blasts. (au) 19 refs.

  9. Expression of prolactin receptor and response to prolactin stimulation of human NK cell lines

    Institute of Scientific and Technical Information of China (English)

    Rui SUN; Ai Ling LI; Hai Ming WEI; Zhi Gang TIAN

    2004-01-01

    We have previously shown a critical role of prolactin (PRL) during maturation and anti-tumor effects of murine natural killer (NK) cells in vitro and in vivo. We extended that study by exploring the ability of human NK cell lines (NK-92 and YT cell) to express PRL receptor (PRL-R) and to respond to PRL stimulation in vitro. Both human NK cell lines constitutively expressed PRL-R on membrane and mRNA transcripts,NK-92 cells contained higher level of PRL-R than YT cells,which correlated to the enhanced capacity of the cells to proliferate and to lyse target cells in response to PRL stimulation in the presence of trace amount of IL-2 or IL-15 in vitro. Two differences between IL-2 and IL-15 in functioning on human NK cells were for the first time observed. PRL synergized with IL-15 to improve proliferation of NK cells in a dose-dependent manner without double peak manifesting like IL-2. Although PRL enhanced the cytotoxicity of IL-2 or IL- 15 activated NK cells,it exerted the function through up-regulating gene expression of perforin without influence of FasL in IL-2-stimulated NK cells,while in IL-15-stimulated NK cells,PRL did the function through up-regulating gene expression of both perforin and FasL but not IFNγ. PRL increased expressions of IL-2Rα on membrane and of IL-2 mRNA in cells,indicating that PRL up-regulated NK cell function by improving positive feedback between IL-2 and IL-2R. The similar results were also observed in network between IL-15 and IL-15R. These data indicate a potential role of PRL in human NK cell modulation.

  10. CD80 (B7-1) expression on human acute myeloid leukaemic cells cultured with GM-CSF, IL-3 and IL-6.

    Science.gov (United States)

    Hicks, C; Keoshkerian, E; Gaudry, L; Lindeman, R

    2001-06-01

    Acute myeloid leukaemia (AML) blasts rarely express the B7 family of co-stimulatory molecules and do not elicit a clinically significant autologous T-lymphocyte anti-tumour response. The aim of this study was the in vitro modification of AML blasts to an antigen-presenting cell phenotype characterised by upregulated expression of the co-stimulatory molecule CD80 (B7-1). Circulating AML cells were induced to undergo partial differentiation in culture with the cytokines IL-3, IL-6 and GM-CSF; they exhibited variable upregulation of CD80 and continued to express MHC class I and II. These cells remained viable to day 20, in contrast with normal peripheral blood mononuclear cells (PBMNC), which did not survive under the culture conditions. In contrast to unmanipulated blasts, cultured leukaemic cells expressed B7-1. Where initial cytogenetic abnormalities were present, they were also seen in flow-sorted CD80-expressing cells after culture in cytokines, indicating their malignant origin. The immunogenic potential of these cultured cells was highlighted by allogeneic and autologous mixed lymphocyte reactions, in which both differentiated, but not unmanipulated, blasts produced expansion of T-lymphocyte numbers. Autologous cytotoxic T-lymphocyte (CTL) assays indicated specific killing of B7-1+ leukaemic cells, which was greatly enhanced after priming of the T-lymphocytes by B7-1+ blasts prior to the CTL assay, then enabling the CTL to lyse both unmanipulated and differentiated leukaemic cells.

  11. Metabolic signatures of Besnoitia besnoiti-infected endothelial host cells and blockage of key metabolic pathways indicate high glycolytic and glutaminolytic needs of the parasite.

    Science.gov (United States)

    Taubert, A; Hermosilla, C; Silva, L M R; Wieck, A; Failing, K; Mazurek, S

    2016-05-01

    Besnoitia besnoiti is an obligate intracellular and emerging coccidian parasite of cattle with a significant economic impact on cattle industry. During acute infection, fast-proliferating tachyzoites are continuously formed mainly in endothelial host cells of infected animals. Given that offspring formation is a highly energy and cell building block demanding process, the parasite needs to exploit host cellular metabolism to meet its metabolic demands. Here, we analyzed the metabolic signatures of B. besnoiti-infected endothelial host cells and aimed to influence parasite proliferation by inhibitors of specific metabolic pathways. The following inhibitors were tested: fluoro 2-deoxy-D-glucose and 2-deoxy-D-glucose (FDG, DG; inhibitors of glycolysis), 6-diazo-5-oxo-L-norleucin (DON; inhibitor of glutaminolysis), dichloroacetate (DCA; inhibitor of pyruvate dehydrogenase kinase which favorites channeling of glucose carbons into the TCA cycle) and adenosine-monophosphate (AMP; inhibitor of ribose 5-P synthesis). Overall, B. besnoiti infections of bovine endothelial cells induced a significant and infection rate-dependent increase of glucose, lactate, glutamine, glutamate, pyruvate, alanine, and serine conversion rates which together indicate a parasite-triggered up-regulation of glycolysis and glutaminolysis. Thus, addition of DON, FDG, and DG into the cultivation medium of B. besnoiti infected endothelial cells led to a dose-dependent inhibition of parasite replication (4 μM DON, 99.5 % inhibition; 2 mM FDG, 99.1 % inhibition; 2 mM DG, 93 % inhibition; and 8 mM DCA, 71.9 % inhibition). In contrast, AMP had no significant effects on total tachyzoite production up to a concentration of 20 mM. Together, these data may open new strategies for the development of therapeutics for B. besnoiti infections.

  12. Red cell indices and discriminant functions in the detection of beta-thalassaemia trait in a population with high prevalence of iron deficiency anaemia.

    Science.gov (United States)

    Madan, N; Sikka, M; Sharma, S; Rusia, U; Kela, K

    1999-01-01

    Red cell indices and discriminant functions were studied in 463 heterozygous beta-thalassaemics (337 without iron deficiency, 126 with iron deficiency) and 195 patients of iron deficiency anaemia (IDA) to ascertain their utility in the detection of betathalassaemia trait (BTT). Majority of traits in both groups had an elevated RBC count (> or = 5.0 x 10(12)/L). The counts were significantly higher than of patients with IDA, only 4.6% of whom had this degree of erythrocytosis. Mean Hb concentration was significantly higher in traits as compared to iron deficient subjects (p < 0.0001). Mean MCV and MCH were significantly (p < 0.0001) lower in traits more so in those with ID as compared to patients of IDA. MCV < 80 fl and MCH < 27 pg were found to be sensitive markers in the detection of traits even in the presence of ID. Of the four discriminant functions studied MCSQ was found to be most sensitive in detection of BTT and it identified 97.9% traits. DF of England and Fraser was most specific for BTT being < 8.4 in only 6.2% patients with IDA. Detection of erythrocytosis especially in the presence of mild anaemia and calculation of discriminant functions derived from red cell indices were found to play an important role in screening for BTT even in the presence of ID and helped identify those patients who required further laboratory evaluation.

  13. Circulating precursor CCR7(lo)PD-1(hi) CXCR5⁺ CD4⁺ T cells indicate Tfh cell activity and promote antibody responses upon antigen reexposure.

    Science.gov (United States)

    He, Jing; Tsai, Louis M; Leong, Yew Ann; Hu, Xin; Ma, Cindy S; Chevalier, Nina; Sun, Xiaolin; Vandenberg, Kirsten; Rockman, Steve; Ding, Yan; Zhu, Lei; Wei, Wei; Wang, Changqi; Karnowski, Alexander; Belz, Gabrielle T; Ghali, Joanna R; Cook, Matthew C; Riminton, D Sean; Veillette, André; Schwartzberg, Pamela L; Mackay, Fabienne; Brink, Robert; Tangye, Stuart G; Vinuesa, Carola G; Mackay, Charles R; Li, Zhanguo; Yu, Di

    2013-10-17

    Follicular B helper T (Tfh) cells support high affinity and long-term antibody responses. Here we found that within circulating CXCR5⁺ CD4⁺ T cells in humans and mice, the CCR7(lo)PD-1(hi) subset has a partial Tfh effector phenotype, whereas CCR7(hi)PD-1(lo) cells have a resting phenotype. The circulating CCR7(lo)PD-1(hi) subset was indicative of active Tfh differentiation in lymphoid organs and correlated with clinical indices in autoimmune diseases. Thus the CCR7(lo)PD-1(hi) subset provides a biomarker to monitor protective antibody responses during infection or vaccination and pathogenic antibody responses in autoimmune diseases. Differentiation of both CCR7(hi)PD-1(lo) and CCR7(lo)PD-1(hi) subsets required ICOS and BCL6, but not SAP, suggesting that circulating CXCR5⁺ helper T cells are primarily generated before germinal centers. Upon antigen reencounter, CCR7(lo)PD-1(hi) CXCR5⁺ precursors rapidly differentiate into mature Tfh cells to promote antibody responses. Therefore, circulating CCR7(lo)PD-1(hi) CXCR5⁺ CD4⁺ T cells are generated during active Tfh differentiation and represent a new mechanism of immunological early memory.

  14. High-risk human papillomavirus infections and overexpression of p53 protein as prognostic indicators in transitional cell carcinoma of the urinary bladder.

    Science.gov (United States)

    Furihata, M; Inoue, K; Ohtsuki, Y; Hashimoto, H; Terao, N; Fujita, Y

    1993-10-15

    Ninety Japanese patients with transitional cell carcinoma of the urinary bladder were investigated for tumor incorporation of DNA for high-risk human papillomavirus (HPV) types 16, 18, and 33 by in situ hybridization with biotinylated DNA probes. In addition, immunohistochemical analysis of p53 protein expression was performed with an antibody to p53 protein. Twenty-eight tumors were positive for HPV DNA, and multiple HPV infection was detected in 17 cases. Positive nuclear staining of cancer cells by the antibody to p53 protein was detected in 32 cases. DNA for HPV 16, 18, and/or 33 and the overexpression of p53 protein were simultaneously observed in 6 tumors by using a mirror section method. The overexpression of p53 protein was frequently detected in invasive and nonpapillary tumors (P infection was more common in noninvasive and papillary tumors (P infection or overexpression of p53 protein may be related to tumor behavior and may indicate a relatively poor prognosis in patients with transitional cell carcinoma.

  15. Agreement and Reliability of Fasted and Oral Glucose Tolerance Test-Derived Indices of Insulin Sensitivity and Beta Cell Function in Boys.

    Science.gov (United States)

    Cockcroft, Emma Joanne; Williams, Craig Anthony; Jackman, Sarah Rebecca; Armstrong, Neil; Barker, Alan R

    2017-06-01

    Assessment of plasma insulin and glucose outcomes is important in paediatric studies aimed at reducing future risk of type 2 diabetes and cardiovascular disease. The aims of this study are to determine the between-method agreement and the day-to-day reliability of fasting and oral glucose tolerance test (OGTT)-derived estimates of insulin sensitivity and β-cell function in healthy boys. Fasting and OGTT assesments of insulin resistance and β-cell function were performed on 28 boys (12.3±2.9 years). Measurements were repeated after 1 week (fasting, n=28) and 1 day (OGTT, n=8). Agreement between estimates of insulin resistance and β-cell function was examined using Pearson's correlation coefficient. Reliability was assessed using change in the mean, Pearson's correlation coefficient, and typical error expressed as a coefficient of variation (CV). The Matsuda index was positively related with QUICKI (r=0.88, P0.05). For reliability, QUICKI had the lowest CV% for the fasting (4.7%) and the Cederholm index for the OGTT (6.4%) estimates. The largest CV% was observed in fasting insulin (30.8%) and insulinogenic index 30' (62.5%). This study highlights differences in between-method agreement and day-to-day reliability for estimates of insulin resistance in youth. The low CV supports the use of the FGIR (fasting) and Cederholm (OGTT) indices in this population. © Georg Thieme Verlag KG Stuttgart · New York.

  16. Progress in tetrazolium salts as indicators of cell viability%四唑盐细胞活力指示剂研究进展

    Institute of Scientific and Technical Information of China (English)

    金诚; 宿连征; 刘汗青; 曹鑫; 谷利军; 王锐

    2012-01-01

    四唑盐类化合物是细胞生物学、细胞与分子毒理学及临床诊断领域用于测定细胞活力的生物指示剂.分别综述了:1)四唑盐及其性质;2)甲臜及其四唑盐的制备方法;3)四唑盐的发展阶段、市场需求以及商品化四唑盐基于比色技术在细胞毒性、抗菌药物的筛选及抗菌药敏实验等方面的应用,并对其发展前景进行了展望.%Tetrazolium salts have become some of the most widely used as indicators in cell biology, molecular cellulartoxicology and clinical diagnosis for measuring cell viability. Three concerning fields are summarized: 1 ) tetrazolium salts and the properties, 2 ) preparation methods of formazans and tetrazolium salts, 3 ) development stages of tetrazolium salts, demand market of tetrazolium salts and the application of commercial tetrazolium salts for antimicrobial susceptibility testing and screening of antimicrobial substances, measuring cell toxicity using conventional colorimetric techniques. At last,the trends of tetrazolium salts are prospected.

  17. CCL22-specific T Cells

    DEFF Research Database (Denmark)

    Martinenaite, Evelina; Munir Ahmad, Shamaila; Hansen, Morten

    2016-01-01

    Tumor cells and tumor-infiltrating macrophages produce the chemokine CCL22, which attracts regulatory T cells (Tregs) into the tumor microenvironment, decreasing anticancer immunity. Here, we investigated the possibility of targeting CCL22-expressing cells by activating specific T cells. We...... analyzed the CCL22 protein signal sequence, identifying a human leukocyte antigen A2- (HLA-A2-) restricted peptide epitope, which we then used to stimulate peripheral blood mononuclear cells (PMBCs) to expand populations of CCL22-specific T cells in vitro. T cells recognizing an epitope derived from...... the signal-peptide of CCL22 will recognize CCL22-expressing cells even though CCL22 is secreted out of the cell. CCL22-specific T cells recognized and killed CCL22-expressing cancer cells. Furthermore, CCL22-specific T cells lysed acute monocytic leukemia cells in a CCL22 expression-dependent manner. Using...

  18. Enhanced Eradication of Lymphoma by Tumor-Specific Cytotoxic T-Cells Secreting an Engineered Tumor-Specific Immunotoxin

    Science.gov (United States)

    2010-06-01

    5% CO2. The level of proliferation was then assessed using an ATP assay (CellTiter-Glo G7570; Promega, Madison, WI). Phenotyping. Cell-surface...trypan blue or an ATP assay (Promega). Cytotoxicity assay. T cell cytotoxic activity was evaluated in a flow cytometry based assay. Target cells...ml PMA and 1µM ionomycin. Cells were collected and lysed, and the lysate analyzed for bioluminescent signal. This revealed that the promoter induced

  19. PARTIAL CHARACTERISATION or A TRYPANOSOME-LYSING ...

    African Journals Online (AJOL)

    2003-11-01

    Nov 1, 2003 ... (trypanolysin) from non-vector insect, Schistocerca gregaria. Design: Laboratory based ..... show how intimate the two molecules function(29). The trypanolysin ... capacity using modern approaches in biotechnology. (17,30).

  20. A study on body mass index, blood pressure, and red blood cell indices in new entering students of the university of isfahan.

    Science.gov (United States)

    Moafi, Alireza; Rahgozar, Soheila; Ghias, Majid; Ahar, Elham Vahdat; Borumand, Amirbahador; Sabbaghi, Amirhosein; Sameti, Amirabass; Hashemi, Mostafa

    2011-10-01

    Obesity and increased blood pressure are identified as risk factors for cardiac and pulmonary disorders. On the other hand, iron deficiency (another preventable disease) is common in adolescence and considered as associated with health impairment. The present study evaluates body mass index (BMI) and its association with blood pressure and hematological indices in freshman students entering the University of Isfahan in 2009. All the 1675 students who entered the University of Isfahan in September 2009 were examined. Height, weight, BMI, blood pressure, hemoglobin (Hb) and red blood cell (RBC) indices of these students were measured. The prevalence of high blood pressure, its association with BMI and the relation between BMI and anemia, iron deficiency and educational achievement were assessed. All participants, including 514 males and 1161 females, went under clinical observations. The average age was 20.7 ± 3.8. year Among the students, 18.2% of males and 20% of females were underweight. High systolic blood pressure was more common in the students with BMI > 25 kg/m(2) (p students' average test scores. High incidence of abnormal BMI in the study population, and its association with systolic blood pressure indicate the importance of nutritional guidelines and counseling programs for freshman students. On the other hand, high incidence of anemia in this population ascertains the necessity of anemia screening programs before academic studies.

  1. Detection of Human Papillomavirus 16-Specific IgG and IgM Antibodies in Patient Sera: A Potential Indicator of Oral Squamous Cell Carcinoma Risk Factor.

    Science.gov (United States)

    Kerishnan, Jesinda P; Gopinath, Subash C B; Kai, Sia Bik; Tang, Thean-Hock; Ng, Helen Lee-Ching; Rahman, Zainal Ariff Abdul; Hashim, Uda; Chen, Yeng

    2016-01-01

    The association between human papillomavirus type 16 (HPV16) and oral cancer has been widely reported. However, detecting anti-HPV antibodies in patient sera to determine risk for oral squamous cell carcinoma (OSCC) has not been well studied. In the present investigation, a total of 206 OSCC serum samples from the Malaysian Oral Cancer Database & Tissue Bank System, with 134 control serum samples, were analyzed by enzyme-linked immunosorbant assay (ELISA) to detect HPV16-specific IgG and IgM antibodies. In addition, nested PCR analysis using comprehensive consensus primers (PGMY09/11 and GP5(+)/6(+)) was used to confirm the presence of HPV. Furthermore, we have evaluated the association of various additional causal factors (e.g., smoking, alcohol consumption, and betel quid chewing) in HPV-infected OSCC patients. Statistical analysis of the Malaysian population indicated that OSCC was more prevalent in female Indian patients that practices betel quid chewing. ELISA revealed that HPV16 IgG, which demonstrates past exposure, could be detected in 197 (95.6%) OSCC patients and HPV16-specific IgM was found in a total of 42 (20.4%) OSCC patients, indicating current exposure. Taken together, our study suggest that HPV infection may play a significant role in OSCC (OR: 13.6; 95% CI: 3.89-47.51) and HPV16-specific IgG and IgM antibodies could represent a significant indicator of risk factors in OSCC patients.

  2. Detection of Human Papillomavirus 16-Specific IgG and IgM Antibodies in Patient Sera: A Potential Indicator of Oral Squamous Cell Carcinoma Risk Factor

    Science.gov (United States)

    Kerishnan, Jesinda P.; Gopinath, Subash C.B.; Kai, Sia Bik; Tang, Thean-Hock; Ng, Helen Lee-Ching; Rahman, Zainal Ariff Abdul; Hashim, Uda; Chen, Yeng

    2016-01-01

    The association between human papillomavirus type 16 (HPV16) and oral cancer has been widely reported. However, detecting anti-HPV antibodies in patient sera to determine risk for oral squamous cell carcinoma (OSCC) has not been well studied. In the present investigation, a total of 206 OSCC serum samples from the Malaysian Oral Cancer Database & Tissue Bank System, with 134 control serum samples, were analyzed by enzyme-linked immunosorbant assay (ELISA) to detect HPV16-specific IgG and IgM antibodies. In addition, nested PCR analysis using comprehensive consensus primers (PGMY09/11 and GP5+/6+) was used to confirm the presence of HPV. Furthermore, we have evaluated the association of various additional causal factors (e.g., smoking, alcohol consumption, and betel quid chewing) in HPV-infected OSCC patients. Statistical analysis of the Malaysian population indicated that OSCC was more prevalent in female Indian patients that practices betel quid chewing. ELISA revealed that HPV16 IgG, which demonstrates past exposure, could be detected in 197 (95.6%) OSCC patients and HPV16-specific IgM was found in a total of 42 (20.4%) OSCC patients, indicating current exposure. Taken together, our study suggest that HPV infection may play a significant role in OSCC (OR: 13.6; 95% CI: 3.89-47.51) and HPV16-specific IgG and IgM antibodies could represent a significant indicator of risk factors in OSCC patients. PMID:27279791

  3. Activation of stress-related signalling pathway in human cells upon SiO2 nanoparticles exposure as an early indicator of cytotoxicity

    LENUS (Irish Health Repository)

    Mohamed, Bashir M

    2011-07-29

    Abstract Background Nanomaterials such as SiO2 nanoparticles (SiO2NP) are finding increasing applications in the biomedical and biotechnological fields such as disease diagnostics, imaging, drug delivery, food, cosmetics and biosensors development. Thus, a mechanistic and systematic evaluation of the potential biological and toxic effects of SiO2NP becomes crucial in order to assess their complete safe applicability limits. Results In this study, human monocytic leukemia cell line THP-1 and human alveolar epithelial cell line A549 were exposed to a range of amorphous SiO2NP of various sizes and concentrations (0.01, 0.1 and 0.5 mg\\/ml). Key biological indicators of cellular functions including cell population density, cellular morphology, membrane permeability, lysosomal mass\\/pH and activation of transcription factor-2 (ATF-2) were evaluated utilizing quantitative high content screening (HCS) approach and biochemical techniques. Despite the use of extremely high nanoparticle concentrations, our findings showed a low degree of cytotoxicity within the panel of SiO2NP investigated. However, at these concentrations, we observed the onset of stress-related cellular response induced by SiO2NP. Interestingly, cells exposed to alumina-coated SiO2NP showed low level, and in some cases complete absence, of stress response and this was consistent up to the highest dose of 0.5 mg\\/ml. Conclusions The present study demonstrates and highlights the importance of subtle biological changes downstream of primary membrane and endocytosis-associated phenomena resulting from high dose SiO2NP exposure. Increased activation of transcription factors, such as ATF-2, was quantitatively assessed as a function of i) human cell line specific stress-response, ii) SiO2NP size and iii) concentration. Despite the low level of cytotoxicity detected for the amorphous SiO2NP investigated, these findings prompt an in-depth focus for future SiO2NP-cell\\/tissue investigations based on the combined

  4. Identification of an Algae-lysing Bacterium of Anabaena flosaquae and Primary Research on Their Relationship%一株水华鱼腥藻溶藻菌的分离鉴定及菌藻关系初探

    Institute of Scientific and Technical Information of China (English)

    叶姜瑜; 钟以蓉; 俞岚; 李文娟

    2011-01-01

    [Objective] The aim was to explore the relationship between alage-lysing bacterium and Anabaena flosaquae so as to provide reference for the control of bloom. [Method] An algae-lysing bacterium strain named S7 was isolated from eutrophic river. The lytic efficiency and performing mode of S7 strain to Anabaena flosaquae was studied. Influence of different environmental factors and the relationship between S7 strain and Anabaena flosaquae was also studied, and then the bacterium strain was physiologically identified. [ Result] More than 90% of Anabaena flosaquae had been removed by 7 d when the volume ratio of medium to algae solution was 30% , the pH was 9 and the temperature was 35 ℃. These results also showed that a mutual inhibit relationship existed between S7 strain and Anabaena flosaquae. The S7 strain killed the algae by indirectly through certain lytic agents in absence of direct contact with the target but by secreting metabolites. Moreover, these lytic agents also had the thermostability. 16S rDNA sequence analysis showed that S7 strain belonged to Chryseobactenum sp.. [Conclusion] The examine Poly-p proved that S7 strain was polyphosphate accumulating bacteria(PAOs) and produced better lytic efficiency.%[目的]研究溶藻特性及菌藻关系,为进一步研究溶藻细菌对水华的治理作用提供帮助.[方法]从富营养化水体中分离得到一株有高效溶藻效果的菌株(S7),研究了其对水华鱼腥藻(Anabaena flosaquae)的抑制效果、作用方式和不同环境因子对溶藻效果的影响,以及菌藻关系,并对菌株进行了菌体Poly-p染色、革兰氏染色和分子鉴定.[结果]菌株投加量为藻波量的30%时,7d叶绿素a的去除率达到90%以上.pH为9、温度35℃下藻的去除率最高.S7菌株与水华鱼腥藻形成竞争共栖的生态关系,并通过分泌溶藻物质间接抑制水华鱼腥藻生长,且该物质具有一定的热稳定性.根据生理生化及16S rDNA序列分析鉴定,S7

  5. Identification of an Algae-lysing Bacterium of Anabaena flosaquae and Primary Research on Their Relationship%一株水华鱼腥藻溶藻菌的分离鉴定及菌藻关系初探

    Institute of Scientific and Technical Information of China (English)

    叶姜瑜; 钟以蓉; 俞岚; 李文娟

    2012-01-01

    [Objective] The aim was to explore the relationship between alage-lysing bacterium and Anabaena flosaquae so as to provide reference for the control of bloom. [Method] An algae-lysing bacterium strain named S7 was isolated from eu- trophic river. The lyric efficiency and performing mode of S7 strain to Anabaena flos- aquae was studied. Influence of different environmental factors and the relationship between S7 strain and Anabaena flosaquae was also studied, and then the bacteri- um strain was physiologically identified. [Result] More than 90% of Anabaena flos- aquae had been removed by 7 d when the volume ratio of medium to algae solu- tion was 30%, the pH was 9 and the temperature was 35 ℃. These results also showed that a mutual inhibit relationship existed between S7 strain and Anabaena flos-aquae. The S7 strain killed the algae by indirectly through certain lyric agents in absence of direct contact with the target but by secreting metabolites. Moreover, these lyric agents also had the thermostability. 16SrDNA sequence analysis showed that S7 strain belonged to Chryseobacterium sp. [Conclusion] The examined Poly-p proved that S7 strain is polyphosphate accumulating bacteria (PAOs) and has better lyric efficiency.%[目的]研究溶藻特性及菌藻关系,为进一步研究溶藻细菌对水华的治理作用提供帮助。[方法]从富营养化水体中分离得到一株有高效溶藻效果的菌株(S7),研究了其对水华鱼腥藻(Anabaenaflosaquae)的抑制效果、作用方式和不同环境因子对溶藻效果的影响,以及菌藻关系,并对菌株进行了菌体Poly—P的染色、革兰氏染色和分子鉴定。【结果]菌株投加量为藻液量的30%时,7d叶绿素a的去除率达到90%以上。pH为9,温度35℃下藻的去除率最高。S7菌株与水华鱼腥藻形成竞争共栖的生态关系,并通过分泌溶藻物质间接抑制水华鱼腥藻生长,且该物质具有一定的热稳定性。根据生

  6. Evaluating 3D printing to solve the sample-to-device interface for LRS and POC diagnostics: example of an interlock meter-mix device for metering and lysing clinical urine samples.

    Science.gov (United States)

    Jue, Erik; Schoepp, Nathan G; Witters, Daan; Ismagilov, Rustem F

    2016-05-21

    This paper evaluates the potential of 3D printing, a semi-automated additive prototyping technology, as a means to design and prototype a sample-to-device interface, amenable to diagnostics in limited-resource settings, where speed, accuracy and user-friendly design are critical components. As a test case, we built and validated an interlock meter-mix device for accurately metering and lysing human urine samples for use in downstream nucleic acid amplification. Two plungers and a multivalve generated and controlled fluid flow through the device and demonstrate the utility of 3D printing to create leak-free seals. Device operation consists of three simple steps that must be performed sequentially, eliminating manual pipetting and vortexing to provide rapid (5 to 10 s) and accurate metering and mixing. Bretherton's prediction was applied, using the bond number to guide a design that prevents potentially biohazardous samples from leaking from the device. We employed multi-material 3D printing technology, which allows composites with rigid and elastomeric properties to be printed as a single part. To validate the meter-mix device with a clinically relevant sample, we used urine spiked with inactivated Chlamydia trachomatis and Neisseria gonorrhoeae. A downstream nucleic acid amplification by quantitative PCR (qPCR) confirmed there was no statistically significant difference between samples metered and mixed using the standard protocol and those prepared with the meter-mix device, showing the 3D-printed device could accurately meter, mix and dispense a human urine sample without loss of nucleic acids. Although there are some limitations to 3D printing capabilities (e.g. dimension limitations related to support material used in the printing process), the advantages of customizability, modularity and rapid prototyping illustrate the utility of 3D printing for developing sample-to-device interfaces for diagnostics.

  7. Definition of transcriptome-based indices for quantitative characterization of chemically disturbed stem cell development: introduction of the STOP-Toxukn and STOP-Toxukk tests.

    Science.gov (United States)

    Shinde, Vaibhav; Hoelting, Lisa; Srinivasan, Sureshkumar Perumal; Meisig, Johannes; Meganathan, Kesavan; Jagtap, Smita; Grinberg, Marianna; Liebing, Julia; Bluethgen, Nils; Rahnenführer, Jörg; Rempel, Eugen; Stoeber, Regina; Schildknecht, Stefan; Förster, Sunniva; Godoy, Patricio; van Thriel, Christoph; Gaspar, John Antonydas; Hescheler, Jürgen; Waldmann, Tanja; Hengstler, Jan G; Leist, Marcel; Sachinidis, Agapios

    2017-02-01

    Stem cell-based in vitro test systems can recapitulate specific phases of human development. In the UKK test system, human pluripotent stem cells (hPSCs) randomly differentiate into cells of the three germ layers and their derivatives. In the UKN1 test system, hPSCs differentiate into early neural precursor cells. During the normal differentiation period (14 days) of the UKK system, 570 genes [849 probe sets (PSs)] were regulated >fivefold; in the UKN1 system (6 days), 879 genes (1238 PSs) were regulated. We refer to these genes as 'developmental genes'. In the present study, we used genome-wide expression data of 12 test substances in the UKK and UKN1 test systems to understand the basic principles of how chemicals interfere with the spontaneous transcriptional development in both test systems. The set of test compounds included six histone deacetylase inhibitors (HDACis), six mercury-containing compounds ('mercurials') and thalidomide. All compounds were tested at the maximum non-cytotoxic concentration, while valproic acid and thalidomide were additionally tested over a wide range of concentrations. In total, 242 genes (252 PSs) in the UKK test system and 793 genes (1092 PSs) in the UKN1 test system were deregulated by the 12 test compounds. We identified sets of 'diagnostic genes' appropriate for the identification of the influence of HDACis or mercurials. Test compounds that interfered with the expression of developmental genes usually antagonized their spontaneous development, meaning that up-regulated developmental genes were suppressed and developmental genes whose expression normally decreases were induced. The fraction of compromised developmental genes varied widely between the test compounds, and it reached up to 60 %. To quantitatively describe disturbed development on a genome-wide basis, we recommend a concept of two indices, 'developmental potency' (D p) and 'developmental index' (D i), whereby D p is the fraction of all developmental genes

  8. Genetic analysis of the Arabidopsis protein kinases MAP3Kε1 and MAP3Kε2 indicates roles in cell expansion and embryo development.

    Science.gov (United States)

    Chaiwongsar, Suraphon; Strohm, Allison K; Su, Shih-Heng; Krysan, Patrick J

    2012-01-01

    MAP3Kε1 and MAP3Kε2 are a pair of Arabidopsis thaliana genes that encode protein kinases related to cdc7p from Saccharomyces cerevisiae. We have previously shown that the map3kε1;map3kε2 double-mutant combination causes pollen lethality. In this study, we have used an ethanol-inducible promoter construct to rescue this lethal phenotype and create map3kε1(-/-);map3kε2(-/-) double-mutant plants in order to examine the function of these genes in the sporophyte. These rescued double-mutant plants carry a yellow fluorescent protein (YFP)-MAP3Kε1 transgene under the control of the alcohol-inducible AlcA promoter from Aspergillus nidulans. The double-mutant plants were significantly smaller and had shorter roots than wild-type when grown in the absence of ethanol treatment. Microscopic analysis indicated that cell elongation was reduced in the roots of the double-mutant plants and cell expansion was reduced in rosette leaves. Treatment with ethanol to induce expression of YFP-MAP3Kε1 largely rescued the leaf phenotypes. The double-mutant combination also caused embryos to arrest in the early stages of development. Through the use of YFP reporter constructs we determined that MAP3Kε1 and MAP3Kε2 are expressed during embryo development, and also in root tissue. Our results indicate that MAP3Kε1 and MAP3Kε2 have roles outside of pollen development and that these genes affect several aspects of sporophyte development.

  9. Towards a Novel Spatially-Resolved Hemolysis Detection Method Using a Fluorescent Indicator and Loaded Ghost Cells: Proof-of-Principle.

    Science.gov (United States)

    Jansen, Sebastian V; Müller, Indra; Kiesendahl, Nicole; Schmitz-Rode, Thomas; Steinseifer, Ulrich

    2015-09-01

    It is of the utmost importance to reduce flow-induced hemolysis in devices such as heart-valve prostheses and blood pumps. Thus, in vitro measurements of hemolysis are performed in order to optimize their design in this regard. However, with existing measurement methods, hemolysis can only be assessed as an integrated value over the complete test-circuit. Currently, there are no spatially-resolved in vitro hemolysis measurement techniques known to the authors that would allow for a determination of the critical regions within a device. In this study, a novel spatially-resolved measurement principle is proposed. Ghost cells (i.e. erythrocytes with a lower hemoglobin concentration) were loaded with a calcium-dicitrato complex, and a fluorescent calcium indicator was suspended in the extracellular medium. Calcium and indicator are separated until the cell membrane ruptures (i.e. hemolysis occurs). In the moment of hemolysis, the two compounds bind to each other and emit a fluorescent signal that can be recorded and spatially-resolved in a setup very similar to a standard Particle Image Velocimetry measurement. A proof-of-principle experiment was performed by intentionally inducing hemolysis in a flow-model with a surfactant. The surfactant-induced hemolysis demonstrated a clear increase of the fluorescent signal compared to that of a negative reference. Furthermore, the signal was spatially restricted to the area of hemolysis. Although further challenges need to be addressed, a successful proof-of-principle for novel spatially-resolved hemolysis detection is presented. This method can contribute to better design optimization of devices with respect to flow-induced hemolysis.

  10. Environmental and human risk assessment of landfill leachate: An integrated approach with the use of cytotoxic and genotoxic stress indices in mussel and human cells

    Energy Technology Data Exchange (ETDEWEB)

    Toufexi, Eirini; Tsarpali, Vasiliki [Section of Animal Biology, Department of Biology, School of Natural Sciences, University of Patras, GR 26500 Patras (Greece); Efthimiou, Ioanna; Vidali, Maria-Sophia; Vlastos, Dimitris [Department of Environmental and Natural Resources Management, University of Patras, 2 Seferi Str., GR 30100 Agrinio (Greece); Dailianis, Stefanos, E-mail: sdailianis@upatras.gr [Section of Animal Biology, Department of Biology, School of Natural Sciences, University of Patras, GR 26500 Patras (Greece)

    2013-09-15

    Highlights: • Landfill leachate poses a threat for aquatic biota and humans. • Leachate induces cytotoxic and oxidative effects on mussel hemocytes. • Increased levels of DNA damage were observed both in vivo and in vitro in hemocytes. • Leachate low doses enhance MN formation in human lymphocyte cultures. • Potential leachate aneugenic activity was detected in human lymphocytes. -- Abstract: The present study investigates leachate hazardous effects on marine biota and human cells, with the use of a battery of assays, both under in vivo and in vitro conditions. According to the results, mussels exposed for 4 days to 0.01 and 0.1% (v/v) of leachate showed increased levels of DNA damage and micronuclei (MN) frequencies in their hemocytes. Similarly, enhanced levels of DNA damage were also observed in hemocytes treated in vitro with relevant concentrations of leachate, followed by a significant enhancement of both superoxide anions (·O{sub 2}{sup −}) and lipid peroxidation products (malondialdehyde/MDA). On the other hand, human lymphocyte cultures treated with such a low concentrations of leachate (0.1, 0.2 and 1%, v/v), showed increased frequencies of MN formation and large MN size ratio, as well as decreased cell proliferation, as indicated by the use of the cytokinesis block micronucleus (CBMN) assay and Cytokinesis Block Proliferation Index (CBPI) respectively. These findings showed the clear-cut genotoxic and cytotoxic effects of leachate on both cellular types, as well as its potential aneugenic activity in human lymphocytes.

  11. Long noncoding RNA PVT1 indicates a poor prognosis of gastric cancer and promotes cell proliferation through epigenetically regulating p15 and p16.

    Science.gov (United States)

    Kong, Rong; Zhang, Er-bao; Yin, Dan-dan; You, Liang-hui; Xu, Tong-peng; Chen, Wen-ming; Xia, Rui; Wan, Li; Sun, Ming; Wang, Zhao-xia; De, Wei; Zhang, Zhi-hong

    2015-04-12

    Mounting evidence indicates that long noncoding RNAs (lncRNAs) could play a pivotal role in cancer biology. However, the overall biological role and clinical significance of PVT1 in gastric carcinogenesis remains largely unknown. Expression of PVT1 was analyzed in 80 GC tissues and cell lines by qRT-PCR. The effect of PVT1 on proliferation was evaluated by MTT and colony formation assays, and cell apoptosis was evaluated by Flow-cytometric analysis. GC cells transfected with shPVT1 were injected into nude mice to study the effect of PVT1 on tumorigenesis in vivo. RIP was performed to confirm the interaction between PVT1 and EZH2. ChIP was used to study the promoter region of related genes. The higher expression of PVT1 was significantly correlated with deeper invasion depth and advanced TNM stage. Multivariate analyses revealed that PVT1 expression served as an independent predictor for overall survival (p = 0.031). Further experiments demonstrated that PVT1 knockdown significantly inhibited the proliferation both in vitro and in vivo. Importantly, we also showed that PVT1 played a key role in G1 arrest. Moreover, we further confirmed that PVT1 was associated with enhancer of zeste homolog 2 (EZH2) and that this association was required for the repression of p15 and p16. To our knowledge, this is the first report showed that the role and the mechanism of PVT1 in the progression of gastric cancer. Together, these results suggest that lncRNA PVT1 may serve as a candidate prognostic biomarker and target for new therapies in human gastric cancer.

  12. A Study on Body Mass Index, Blood Pressure, and Red Blood Cell Indices in New Entering Students of the University of Isfahan

    Science.gov (United States)

    Moafi, Alireza; Rahgozar, Soheila; Ghias, Majid; ahar, Elham Vahdat; Borumand, Amirbahador; Sabbaghi, Amirhosein; Sameti, Amirabass; Hashemi, Mostafa

    2011-01-01

    Objectives: Obesity and increased blood pressure are identified as risk factors for cardiac and pulmonary disorders. On the other hand, iron deficiency (another preventable disease) is common in adolescence and considered as associated with health impairment. The present study evaluates body mass index (BMI) and its association with blood pressure and hematological indices in freshman students entering the University of Isfahan in 2009. Methods: All the 1675 students who entered the University of Isfahan in September 2009 were examined. Height, weight, BMI, blood pressure, hemoglobin (Hb) and red blood cell (RBC) indices of these students were measured. The prevalence of high blood pressure, its association with BMI and the relation between BMI and anemia, iron deficiency and educational achievement were assessed. Results: All participants, including 514 males and 1161 females, went under clinical observations. The average age was 20.7 ± 3.8. year Among the students, 18.2% of males and 20% of females were underweight. High systolic blood pressure was more common in the students with BMI > 25 kg/m2 (p < 0.001). Anemia was seen in 8.7% of females. In males, however, a relation between anemia frequency and BMI < 18.5 kg/m2 was more distinct (p = 0.002). There was no association between anemia and students’ average test scores. Conclusions: High incidence of abnormal BMI in the study population, and its association with systolic blood pressure indicate the importance of nutritional guidelines and counseling programs for freshman students. On the other hand, high incidence of anemia in this population ascertains the necessity of anemia screening programs before academic studies. PMID:22174970

  13. Primary WWOX phosphorylation and JNK activation during etoposide induces cytotoxicity in HEK293 cells

    Directory of Open Access Journals (Sweden)

    M Jamshidiha

    2010-06-01

    Full Text Available "n  "nBackground and the purpose of the study: Etoposide is an antineoplastic agent used in multiple cancers. It is known that etoposide induce cell death via interaction with topoisomerase II; however, the etopoisde cellular response is poorly understood. Upon etoposide induced DNA damage, many stress signaling pathways including JNK are activated. In response to DNA damage, it has been shown that WWOX, a recently introduced tumor suppressor, can be activated. In this study the activation of WWOX and JNK and their interaction following etoposide treatment were evaluated. "nMaterials and Methods:HEK293 cells treated with etoposide were lysed in a time course manner. The whole cell lysates were used to evaluate JNK and WWOX activation pattern using Phospho specific antibodies on western blots. The viability of cells treated with etoposide, JNK specific inhibitor and their combination was examined using MTT assay. "nResults:Findings of this study indicate that WWOX and JNK are activated in a simultaneous way in response to DNA damage. Moreover, JNK inhibition enhances etoposide induced cytotoxicity in HEK293. "nConclusion:Taken together, our results indicate that etoposide induces cytotoxicity and WWOX phosphorylation and the cytotoxicty is augmented by blocking JNK pathway.

  14. The effect of microgravity on the in vitro NK cell function during six International Space Station Missions

    Science.gov (United States)

    Buravkova, L. B.; Grigorieva, V.; Rykova, M. P.

    2007-09-01

    The level of natural killer (NK) cytotoxic activity was measured during co-cultivation of human lymphocytes and target cells (K-562) in microgravity. Flight experiments were carried out using special instrumentation, the "Fibroblast-1 " cassettes, in the frame of Russian scientific program during six ISS missions. Lymphocyte suspensions from human venous blood were used in experiments during short-term flights on six ISS missions (7-12). Russian space crew members performed the experiments after Soyuz docking. The first step was mixing lymphocytes and3H-labeled K-562 cells and their incubation at 37°C during 24 hs; the second step was filtration of the cell suspension. The frozen medium and filters were analyzed for the cytokine level and cytotoxic activity after landing. It was found that lymphocytes with different basal levels of cytotoxic activity kept the ability of recognizing and lysing malignant cells. In microgravity, cytotoxity increased to 160% of the basal levels. Donor individual features modulated the magnitude of the increase. The measurement of interleukin levels (TNF-α, IL-1, IL-2) in medium showed that synthesis of TNF-α increased during cell co-cultivation in microgravity. The level of IL-2 was very low inflight and ground control samples. The production of IL-1 by lymphocytes decreased after in-flight incubation. The results indicate that microgravity did not disturb the cytotoxic function of immune cells in vitro during 24 h incubation with specific target cells.

  15. DNAM-1 mediates epithelial cell-specific cytotoxicity of aberrant intraepithelial lymphocyte lines from refractory celiac disease type II patients.

    Science.gov (United States)

    Tjon, Jennifer M-L; Kooy-Winkelaar, Yvonne M C; Tack, Greetje J; Mommaas, A Mieke; Schreurs, Marco W J; Schilham, Marco W; Mulder, Chris J; van Bergen, Jeroen; Koning, Frits

    2011-06-01

    In refractory celiac disease (RCD), intestinal epithelial damage persists despite a gluten-free diet. Characteristic for RCD type II (RCD II) is the presence of aberrant surface TCR-CD3(-) intraepithelial lymphocytes (IELs) that can progressively replace normal IELs and eventually give rise to overt lymphoma. Therefore, RCD II is considered a malignant condition that forms an intermediate stage between celiac disease (CD) and overt lymphoma. We demonstrate in this study that surface TCR-CD3(-) IEL lines isolated from three RCD II patients preferentially lyse epithelial cell lines. FACS analysis revealed that DNAM-1 was strongly expressed on the three RCD cell lines, whereas other activating NK cell receptors were not expressed on all three RCD cell lines. Consistent with this finding, cytotoxicity of the RCD cell lines was mediated mainly by DNAM-1 with only a minor role for other activating NK cell receptors. Furthermore, enterocytes isolated from duodenal biopsies expressed DNAM-1 ligands and were lysed by the RCD cell lines ex vivo. Although DNAM-1 on CD8(+) T cells and NK cells is known to mediate lysis of tumor cells, this study provides, to our knowledge, the first evidence that (pre)malignant cells themselves can acquire the ability to lyse epithelial cells via DNAM-1. This study confirms previous work on epithelial lysis by RCD cell lines and identifies a novel mechanism that potentially contributes to the gluten-independent tissue damage in RCD II and RCD-associated lymphoma.

  16. Modulation of Oxidative Stress by Gamma-Glutamylcysteine (GGC) and Conjugated Linoleic Acid (CLA) Isomer Mixture in Human Umbilical Vein Endothelial Cells

    Science.gov (United States)

    2012-04-02

    treatment and pooled. Then, cells were suspended and lysed with a hypotonic buffer and 10% (w/v) Nonidet P - 40 . After spinning, the cell pellet was re...imately 40 % cell death was microscopically observed in cells incubated with GGC and 100 lmol/L CLA. Despite lower levels of GSH, treatment with GGC... p < 0.05 were considered to be sta- tistically significant. All results were expressed as mean ± standard deviation. 3. Results 3.1. Cell viability

  17. The modified method of two-step differential extraction of sperm and vaginal epithelial cell DNA from vaginal fluid mixed with semen.

    Science.gov (United States)

    Yoshida, K; Sekiguchi, K; Mizuno, N; Kasai, K; Sakai, I; Sato, H; Seta, S

    1995-03-21

    This investigation was undertaken as an efficient method for isolating sperm DNA from a mixed fluid sample which contains vaginal epithelial cells in a greater amount. The modified method of the two-step differential extraction procedure was found to be suitable for separating sperm DNA and vaginal epithelial cell DNA from the mixed stains. As the first step of digestion, vaginal epithelial cells in the mixed stains were lysed with Proteinase K and SDS, and sperm heads remaining in the lysed solution were collected by centrifugation. As the second step digestion, the sperm heads were lysed with the buffer containing Proteinase K, SDS and DTT as reducing agent. DNA fractions extracted from the two lysed solutions were enriched, one with sperm DNA and the other with vaginal epithelial cell DNA. MCT118(D1S80), ApoB VNTR and HLADQ alpha types of sperm DNA were detected and were confirmed by matching with corresponding male blood DNA. In the case of vaginal secretion mixed with semen of two males, the mixture of MCT118 types of the two males was detected in sperm DNA fraction.

  18. Dual optical recordings for action potentials and calcium handling in induced pluripotent stem cell models of cardiac arrhythmias using genetically encoded fluorescent indicators.

    Science.gov (United States)

    Song, LouJin; Awari, Daniel W; Han, Elizabeth Y; Uche-Anya, Eugenia; Park, Seon-Hye E; Yabe, Yoko A; Chung, Wendy K; Yazawa, Masayuki

    2015-05-01

    Reprogramming of human somatic cells to pluripotency has been used to investigate disease mechanisms and to identify potential therapeutics. However, the methods used for reprogramming, in vitro differentiation, and phenotyping are still complicated, expensive, and time-consuming. To address the limitations, we first optimized a protocol for reprogramming of human fibroblasts and keratinocytes into pluripotency using single lipofection and the episomal vectors in a 24-well plate format. This method allowed us to generate multiple lines of integration-free and feeder-free induced pluripotent stem cells (iPSCs) from seven patients with cardiac diseases and three controls. Second, we differentiated human iPSCs derived from patients with Timothy syndrome into cardiomyocytes using a monolayer differentiation method. We found that Timothy syndrome cardiomyocytes showed slower, irregular contractions and abnormal calcium handling compared with the controls. The results are consistent with previous reports using a retroviral method for reprogramming and an embryoid body-based method for cardiac differentiation. Third, we developed an efficient approach for recording the action potentials and calcium transients simultaneously in control and patient cardiomyocytes using genetically encoded fluorescent indicators, ArcLight and R-GECO1. The dual optical recordings enabled us to observe prolonged action potentials and abnormal calcium handling in Timothy syndrome cardiomyocytes. We confirmed that roscovitine rescued the phenotypes in Timothy syndrome cardiomyocytes and that these findings were consistent with previous studies using conventional electrophysiological recordings and calcium imaging with dyes. The approaches using our optimized methods and dual optical recordings will improve iPSC applicability for disease modeling to investigate mechanisms underlying cardiac arrhythmias and to test potential therapeutics.

  19. The Value of Erythrocyte Indices and Red Cell Volume Distribution Width in Differential Diagnosis of Iron Deficiency Anemia and Anemia of Chronic Disease

    Directory of Open Access Journals (Sweden)

    Abdullah Altıntaş

    2007-01-01

    Full Text Available Iron deficiency anemia (IDA and anemia of chronic disease are the most commonly confused anemias. We investigate the diagnostic value of erythrocyte indices, red cell volume distribution width, and serum ferritin levels to make differential diagnosis of anemia in controls and anemic patients.Iron deficiency anemia (44 patients, anemia of chronic disease (41, IDA with anemia of chronic disease (17 and control (50 groups were compared. We performed serum ferritin, CBC, and sedimentation rate in all patientsand bone marrow aspiration in patients with anemia of chronic disease.Although mean cell volume (MCV and mean corpuscular hemoglobin (MCH are low in IDA and anemia of chronic disease, it is much striking in the former one (p<0.001, p<0.001. Only 7.3% of patients with anemia of chronic disease had a MCV<70 fL and MCH<24 pg, 90.0% of patients with IDA were below that cut-off point. Serum ferritin means were in patients with IDA and anemia of chronic disease were 4.6±3.3 and 489.6 ±519.9 ng/ml, respectively.The probability of IDA is low when RDW is normal in microcytic anemias. RDW is high in half of patients with anemia of chronic disease. Reference values for ferritin must be changed in patients with anemia of chronic disease and IDA. If serum ferritin is 57.6-146.4 ng/ml anemia of chronic disease and iron deficiency must ruled out by other diagnostic tests.

  20. Receptor activity on some mesenchymal cells in CNS of normal rabbits. Indications of the monocytic origin of intracerebral perivascular cells, epiplexus cells and mononuclear phagocytes in the subarachnoid space.

    Science.gov (United States)

    Oehmichen, M

    1976-01-01

    The surface receptor activity for various cell types within rabbit CNS was investigated. Sheep red blood cells (E) used as markers were washed, sensitized with the IgG-fraction of E-antibodies (EA) or additionally coated with complement (EAC) and incubated with CNS cells. The inhibitory effect produced by the addition of soluble IgG was investigated. Incubation (1 h) with red cells was undertaken in three ways: 1. Rabbit leptomeninx was stripped and incubated. 2. Rabbits were killed, the brain was perfused with warm buffer and red cell complexes were then injected intracerebrally, intrathecally and intraventricularly into the perfused brains. 3. E, EA and EAC were injected intracerebrally, intrathecally and intra-ventricularly into living rabbits. Using these methods, receptor sites for IgG and complement of mononuclear cells from the subarachnoid space, epiplexus cells and perivascular cells from intracerebral vessels could be demonstrated. In other areas of the body, these receptors have been demonstrated using similar methods only on cells from the monocyte-macrophage series. The common derivation of these three cell types from the monocytes as well as their comparable function within the immune response is discussed.

  1. Cell death biomarker M65 is a useful indicator of liver inflammation and fibrosis in chronic hepatitis B: A cross-sectional study of diagnostic accuracy.

    Science.gov (United States)

    Wei, Xinhuan; Wei, Hongshan; Lin, Wei; Hu, Zhongjie; Zhang, Jing

    2017-05-01

    Cell death markers, M65 and M30, have been suggested to be sensitive markers of liver inflammation and fibrosis in nonalcoholic fatty liver disease and chronic hepatitis C. Our aim was to investigate whether these markers were useful in diagnosing liver inflammation and fibrosis in chronic hepatitis B (CHB).We examined 186 patients with CHB; 18 sex- and age-matched healthy subjects were controls. The blood samples were collected from CHB patients within 1 week before or after liver biopsy. According to METAVIR score system, liver inflammation was graded from A0 to A3, and fibrosis from F0 to F4.Serum M65 and M30 levels were in parallel with the grades of liver inflammation. M65, not M30, increased significantly in patients with severe inflammation and normal alanine aminotransferase. M65 is one of the independent predictors of severe liver inflammation (≥A2). The levels of M65 and M30 levels significantly increased in parallel with the degree of inflammation in F1 patients, whereas they showed no statistical difference between different stages of fibrosis in A1 patients.Serum M65 is a useful indicator of liver inflammation in CHB patients. Serum M65, not M30, is valuable in the grading of liver fibrosis.

  2. Preoperative red cell distribution width: Not a useful prognostic indicator for 30-day mortality in patients who undergo major- or ultra-major noncardiac surgery

    Directory of Open Access Journals (Sweden)

    Yik-Nang Cheung

    2016-01-01

    Full Text Available Background: Red cell distribution width (RDW has been shown to be associated with mortality in cardiac surgical patients. This study investigates the association of RDW with the 30-day mortality for those patients who undergo major- or ultra-major noncardiac surgery. Methods: Patients who received major- or ultra-major noncardiac surgery between July 2012 and May 2013 were included in the study and patients those with preoperative hemoglobin 13.35% (P = 0.025, odds ratio [OR]: 1.52, INR (P = 0.008, OR: 4.49, albumin level (P < 0.001, OR: 1.10, use of antihypertensives (P = 0.001, OR: 1.82, and preoperative pulse rate (P = 0.006, OR: 1.02 independently predicted the 30-day mortality. However, the area under receiver operating characteristic curve for the prediction of 30-day mortality using RDW was only 0.614. Conclusions: Although preoperative RDW independently predicted 30-day mortality in patients who underwent major- or ultra-major noncardiac surgery, it may not serve as an influential prognostic indicator in view of its low sensitivity and specificity.

  3. Fatty acid content, health and risk indices, physicochemical composition, and somatic cell counts of milk from organic and conventional farming systems in tropical south-eastern Mexico.

    Science.gov (United States)

    Delgadillo-Puga, Claudia; Sánchez-Muñoz, Bernardo; Nahed-Toral, José; Cuchillo-Hilario, Mario; Díaz-Martínez, Margarita; Solis-Zabaleta, Roman; Reyes-Hernández, Aurora; Castillo-Domíguez, Rosa Maria

    2014-06-01

    Organic agriculture and livestock farming is claimed to promote animal welfare and can offer animal products with better hygienic-sanitary quality, based on principles of health, ecology, fairness, and care. However, no clear advantages of organic milk (OM) versus conventional milk (CM) from tropical conditions are available. The aims of the study were to determine fatty acid profile, health-promoting (HPI) and thrombogenic (TI) indices, physicochemical composition, and somatic cell counts (SCC) of OM and CM in tropical south-eastern Mexico. Female cross-breed cows (400-600 kg) were employed. CM had larger values of saturated fatty acids (SFA) and polyunsaturated fatty acid (PUFA) (63.6 %; 4.57 %) than OM (61.48 %; 4.22 %), while OM resulted in a larger value of monounsaturated fatty acid (MUFA) (34.3 %) than CM (31.7 %). HPI and TI showed that OM was more favorable than CM. Milk production and physicochemical composition (PC) as well as density had no significant difference, while SCC was significantly lower in OM than in CM on a monthly basis. These results showed that OM promotes a healthful and balanced diet, and is already produced by sustainable ecologic technologies employing traditional agrosilvopastoral management, which is more environmentally friendly and promotes ecological resilience.

  4. Repeatability of measurements of packed cell volume and egg count as indicators of endoparasite load and their relationship with sheep productivity.

    Science.gov (United States)

    Bekele, T; Kasali, O B; Rege, J E

    1991-12-01

    Monthly measurements of packed cell volume (PCV) and nematode and trematode eggs per gram (EPG) were made in Ethiopian highland sheep at Debre Berhan, Dejen, Deneba, Tulu Meko and Wereilu from June 1988 to December 1989. High frequencies of low PCV, high nematode EPG and high trematode EPG were found at Tulu Meko. Among the productivity traits examined, body condition scores and live-weights were significantly (P less than 0.05) associated with differences in PCV and nematode and trematode EPG levels at most sites. The lambing interval was, however, not significantly (P greater than 0.05) affected by these variables. Monthly repeatabilities of PCV, body weight and body condition scores were 0.44 +/- 0.01, 0.71 +/- 0.01 and 0.35 +/- 0.01, respectively, while those of nematode (0.09 +/- 0.01) and trematode EPGs (0.20 +/- 0.02) were much lower. The high repeatability for PCV indicates that it was less affected by the variable factors influencing egg output, and hence it could be utilized in conjunction with nematode and trematode EPG levels for endoparasite monitoring. Repeatability of the lambing interval across parities was 0.43 +/- 0.14.

  5. Phenylpropanoid Defences in Nicotiana tabacum Cells: Overlapping Metabolomes Indicate Common Aspects to Priming Responses Induced by Lipopolysaccharides, Chitosan and Flagellin-22.

    Directory of Open Access Journals (Sweden)

    Msizi I Mhlongo

    Full Text Available Plants have evolved both constitutive and inducible defence strategies to cope with different biotic stimuli and stresses. Exposure of a plant to a challenging stress can lead to a primed state that allows it to launch a more rapid and stronger defence. Here we applied a metabolomic approach to study and compare the responses induced in Nicotiana tabacum cells by microbe-associated molecular pattern (MAMP molecules, namely lipopolysaccharides (LPS, chitosan (CHT and flagellin-22 (FLG22. Early response metabolites, extracted with methanol, were analysed by UHPLC-MS/MS. Using multivariate statistical tools the metabolic profiles induced by these elicitors were analysed. In the metabolic fingerprint of these agents a total of 19 cinnamic acid derivatives conjugated to quinic acids (chlorogenic acids, shikimic acid, tyramine, polyamines or glucose were found as discriminant biomarkers. In addition, treatment with the phytohormones salicylic acid (SA, methyljasmonic acid (MJ and abscisic acid (ABA resulted in differentially-induced phenylpropanoid pathway metabolites. The results indicate that the phenylpropanoid pathway is activated by these elicitors while hydroxycinnamic acid derivatives are commonly associated with the metabolic response to the MAMPs, and that the activated responses are modulated by both SA and MJ, with ABA not playing a role.

  6. A combinational therapy of EGFR-CAR NK cells and oncolytic herpes simplex virus 1 for breast cancer brain metastases.

    Science.gov (United States)

    Chen, Xilin; Han, Jianfeng; Chu, Jianhong; Zhang, Lingling; Zhang, Jianying; Chen, Charlie; Chen, Luxi; Wang, Youwei; Wang, Hongwei; Yi, Long; Elder, J Bradley; Wang, Qi-En; He, Xiaoming; Kaur, Balveen; Chiocca, E Antonio; Yu, Jianhua

    2016-05-10

    Breast cancer brain metastases (BCBMs) are common in patients with metastatic breast cancer and indicate a poor prognosis. These tumors are especially resistant to currently available treatments due to multiple factors. However, the combination of chimeric antigen receptor (CAR)-modified immune cells and oncolytic herpes simplex virus (oHSV) has not yet been explored in this context. In this study, NK-92 cells and primary NK cells were engineered to express the second generation of EGFR-CAR. The efficacies of anti-BCBMs of EGFR-CAR NK cells, oHSV-1, and their combination were tested in vitro and in a breast cancer intracranial mouse model. In vitro, compared with mock-transduced NK-92 cells or primary NK cells, EGFR-CAR-engineered NK-92 cells and primary NK cells displayed enhanced cytotoxicity and IFN-γ production when co-cultured with breast cancer cell lines MDA-MB-231, MDA-MB-468, and MCF-7. oHSV-1 alone was also capable of lysing and destroying these cells. However, a higher cytolytic effect of EGFR-CAR NK-92 cells was observed when combined with oHSV-1 compared to the monotherapies. In the mice intracranially pre-inoculated with EGFR-expressing MDA-MB-231 cells, intratumoral administration of either EGFR-CAR-transduced NK-92 cells or oHSV-1 mitigated tumor growth. Notably, the combination of EGFR-CAR NK-92 cells with oHSV-1 resulted in more efficient killing of MDA-MB-231 tumor cells and significantly longer survival of tumor-bearing mice when compared to monotherapies. These results demonstrate that regional administration of EGFR-CAR NK-92 cells combined with oHSV-1 therapy is a potentially promising strategy to treat BCBMs.

  7. High frequency of cytolytic 21-Hydroxylase specific CD8+ T cells in autoimmune Addison’s disease patients1

    Science.gov (United States)

    Dawoodji, Amina; Chen, Ji-Li; Shepherd, Dawn; Dalin, Frida; Tarlton, Andrea; Alimohammadi, Mohammad; Penna-Martinez, Marissa; Meyer, Gesine; Mitchell, Anna L; Gan, Earn H; Bratland, Eirik; Bensing, Sophie; Husebye, Eystein; Pearce, Simon H.; Badenhoop, Klaus; Kämpe, Olle; Cerundolo, Vincenzo

    2016-01-01

    The mechanisms behind the destruction of the adrenal glands in autoimmune Addison’s disease remain unclear. Autoantibodies against steroid 21-hydroxylase, an intracellular key enzyme of the adrenal cortex, are found in over 90% of patients, but these autoantibodies are not thought to mediate the disease. Here we demonstrate highly frequent 21-hydroxylase specific T cells detectable in 20 patients with Addison’s disease. Using overlapping 18aa peptides spanning the full length of 21-hydroxylase, we identified immunodominant CD8+ and CD4+ T cell responses in a large proportion of Addison’s patients both ex-vivo and after in-vitro culture of peripheral blood lymphocytes up to 20 years after diagnosis. In a large proportion of patients, CD8+ 21-hydroxylase specific T cells and CD4+ 21-hydroxylase specific T cells were very abundant and detectable in ex-vivo assays. HLA class-I tetramer-guided isolation of 21-hydroxylase specific CD8+ T cells showed their ability to lyse 21-hydroxylase positive target cells, consistent with a potential mechanism for disease pathogenesis. These data indicate strong cytotoxic T lymphocyte responses to 21-hydroxylase often occur in-vivo, and that reactive cytotoxic T lymphocytes have substantial proliferative and cytolytic potential. These results have implications for earlier diagnosis of adrenal failure and ultimately a potential target for therapeutic intervention and induction of immunity against adrenal cortex cancer. PMID:25063864

  8. Renal cell carcinoma-infiltrating natural killer cells express differential repertoires of activating and inhibitory receptors and are inhibited by specific HLA class I allotypes.

    Science.gov (United States)

    Schleypen, Julia S; Von Geldern, Marion; Weiss, Elisabeth H; Kotzias, Nicole; Rohrmann, Karl; Schendel, Dolores J; Falk, Christine S; Pohla, Heike

    2003-10-10

    Among tumor-infiltrating lymphocytes (TILs) directly isolated from renal cell carcinomas (RCCs), we found substantial numbers of natural killer (NK) cells in most tumor tissues. They could be identified reliably in situ with an antibody directed against the activating receptor (AR) NKp46 that is exclusively expressed by all NK cells. NK-enriched TILs (NK-TILs) showed cytotoxicity against major histocompatibility complex (MHC) class I-negative cell lines. The ability to detect lysis of target cells was dependent on the percentage of NK cells within the TILs, and cytotoxicity was only observed after overnight activation with low-dose interleukin-2 (IL-2). Infiltrating NK cells were found to express various inhibitory receptors (IRs); among these the CD94/NKG2A receptor complex was overrepresented compared to the autologous peripheral blood mononuclear cell (PBMC) population. Other IRs were underrepresented, indicating that NK subpopulations vary in their tumor-infiltrating capacity. IRs expressed by NK-TILs are functional since receptor engagement with MHC class I ligands presented by human leukocyte antigen (HLA)-transfected target cell lines was able to inhibit NK-mediated cytotoxicity. NK-TILs were also able to lyse autologous or allogeneic tumor cell lines in vitro. This activity correlated with low HLA class I surface expression since lysis could be inhibited by interferon (IFN)-gamma-expressing RCC transductants that displayed a higher surface density of HLA class I molecules. Therefore, NK cells infiltrating tumor tissues have an inherent ability to recognize transformed cells, but they require cytokine activation and are sensitive to inhibition by IR ligands.

  9. Role of protein kinase C in TBT-induced inhibition of lytic function and MAPK activation in human natural killer cells.

    Science.gov (United States)

    Abraha, Abraham B; Rana, Krupa; Whalen, Margaret M

    2010-11-01

    Human natural killer (NK) cells are lymphocytes that destroy tumor and virally infected cells. Previous studies have shown that exposure of NK cells to tributyltin (TBT) greatly diminishes their ability to destroy tumor cells (lytic function) while activating mitogen-activated protein kinases (MAPK) (p44/42, p38, and JNK) in NK cells. The signaling pathway that regulates NK lytic function appears to include activation of protein kinase C(PKC) as well as MAPK activity. TBT-induced activation of MAPKs would trigger a portion of the NK lytic signaling pathway, which would then leave the NK cell unable to trigger this pathway in response to a subsequent encounter with a target cell. In the present study we evaluated the involvement of PKC in inhibition of NK lysis of tumor cells and activation of MAPKs caused by TBT exposure. TBT caused a 2–3-fold activation of PKC at concentrations ranging from 50 to 300 nM (16–98 ng/ml),indicating that activation of PKC occurs in response to TBT exposure. This would then leave the NK cell unable to respond to targets. Treatment with the PKC inhibitor, bisindolylmaleimide I, caused an 85% decrease in the ability of NK cells to lyse tumor cells, validating the involvement of PKC in the lytic signaling pathway. The role of PKC in the activation of MAPKs by TBT was also investigated using bisindolylmaleimide I. The results indicated that, in NK cells where PKC activation was blocked, there was no activation of the MAPK, p44/42 in response to TBT.However, TBT-induced activation of the MAPKs, p38 and JNK did not require PKC activation. These results indicate the pivotal role of PKC in the TBT-induced loss of NK lytic function including activation of p44/42 by TBT in NK cells.

  10. Efficient analysis of a small number of cancer cells at the single-cell level using an electroactive double-well array.

    Science.gov (United States)

    Kim, Soo Hyeon; Fujii, Teruo

    2016-07-01

    Analysis of the intracellular materials of a small number of cancer cells at the single-cell level is important to improve our understanding of cellular heterogeneity in rare cells. To analyze an extremely small number of cancer cells (less than hundreds of cells), an efficient system is required in order to analyze target cells with minimal sample loss. Here, we present a novel approach utilizing an advanced electroactive double-well array (EdWA) for on-chip analysis of a small number of cancer cells at the single-cell level with minimal loss of target cells. The EdWA consisted of cell-sized trap-wells for deterministic single-cell trapping using dielectrophoresis and high aspect ratio reaction-wells for confining the cell lysates extracted by lysing trapped single cells via electroporation. We demonstrated a highly efficient single-cell arraying (a cell capture efficiency of 96 ± 3%) by trapping diluted human prostate cancer cells (PC3 cells). On-chip single-cell analysis was performed by measuring the intracellular β-galactosidase (β-gal) activity after lysing the trapped single cells inside a tightly enclosed EdWA in the presence of a fluorogenic enzyme substrate. The PC3 cells showed large cell-to-cell variations in β-gal activity although they were cultured under the same conditions in a culture dish. This simple and effective system has great potential for high throughput single-cell analysis of rare cells.

  11. Potential therapeutic strategy for gastric cancer peritoneal metastasis by NKG2D ligands-specific T cells

    Directory of Open Access Journals (Sweden)

    Liu XQ

    2015-10-01

    Full Text Available Xianqiang Liu,1 Meili Sun,2 Shui Yu,3 Kai Liu,4 Xirui Li,5 Huan Shi6 1Department of Breast and Thyroid Surgery, 2Department of Oncology, Jinan Central Hospital Affiliated to Shandong University, 3Department of Radiation Oncology, 4Department of Gastrointestinal Surgery, 5Medical Department, 6Department of Oncology, Shandong Cancer Hospital and Institute, Jinan, Shandong, People’s Republic of China Purpose: Despite advancements in its treatment, gastric cancer continues to be one of the leading causes of cancer deaths worldwide. Adoptive transfer of chimeric antigen receptor-modified T cells is a promising antitumor therapy for many cancers. The purpose of this study was to construct a chimeric receptor linking the extracellular domain of NKG2D to the CD28 and CD3zeta chain intracellular domains to target gastric cancers that expressed NKG2D ligands.Methods: Expression of NKG2D ligands including MICA, MICB, and ULBP1–3 in a gastric cancer cell line and primary gastric cancer cells from ascites samples were analyzed using flow cytometry. Co-culture experiments were performed by incubating chNKG2D T cells with gastric cancer cell lines and with primary human gastric cancer cells isolated from ascites and by measuring cytokine and chemokine release and cytotoxicity.Results: Gastric cancer cell lines and ascites-derived primary human gastric cancer cells expressed high levels of MICA, MICB, and ULBP2. ChNKG2D T cells secreted proinflammatory cytokines and chemokines when cultured with these cancer cells. In addition, chNKG2D T cells lysed gastric cancer cell lines and the ascites-derived primary human gastric cancer cells.Conclusion: These data indicate that treatment with chNKG2D-expressing T cells is a potential immunotherapy for gastric cancer with peritoneal metastasis. Keywords: chimeric antigen receptor, T cells, immunotherapy

  12. MicroRNA-224 is Readily Detectable in Urine of Individuals with Diabetes Mellitus and is a Potential Indicator of Beta-Cell Demise

    Directory of Open Access Journals (Sweden)

    Siobhán Bacon

    2015-06-01

    Full Text Available MicroRNA (miRNA are a class of non-coding, 19–25 nucleotide RNA critical for network-level regulation of gene expression. miRNA serve as paracrine signaling molecules. Using an unbiased array approach, we previously identified elevated levels of miR-224 and miR-103 to be associated with a monogenic form of diabetes; HNF1A-MODY. miR-224 is a novel miRNA in the field of diabetes. We sought to explore the role of miR-224 as a potential biomarker in diabetes, and whether such diabetes-associated-miRNA can also be detected in the urine of patients. Absolute levels of miR-224 and miR-103 were determined in the urine of n = 144 individuals including carriers of a HNF1A mutation, participants with type 1 diabetes mellitus (T1DM, type 2 diabetes mellitus (T2DM and normal controls. Expression levels were correlated with clinical and biochemical parameters. miR-224 was significantly elevated in the urine of carriers of a HNF1A mutation and participants with T1DM. miR-103 was highly expressed in urine across all diabetes cohorts when compared to controls. For both miR-224 and-103, we found a significant correlation between serum and urine levels (p < 0.01. We demonstrate that miRNA can be readily detected in the urine independent of clinical indices of renal dysfunction. We surmise that the differential expression levels of miR-224 in both HNF1A-MODY mutation carriers and T1DM may be an attempt to compensate for beta-cell demise.

  13. Expression profile of genes as indicators of developmental competence and quality of in vitro fertilization and somatic cell nuclear transfer bovine embryos.

    Directory of Open Access Journals (Sweden)

    Maria Jesús Cánepa

    Full Text Available Reproductive biotechnologies such as in vitro fertilization (IVF and somatic cell nuclear transfer (SCNT enable improved reproductive efficiency of animals. However, the birth rate of in vitro-derived embryos still lags behind that of their in vivo counterparts. Thus, it is critical to develop an accurate evaluation and prediction system of embryo competence, both for commercial purposes and for scientific research. Previous works have demonstrated that in vitro culture systems induce alterations in the relative abundance (RA of diverse transcripts and thus compromise embryo quality. The aim of this work was to analyze the RA of a set of genes involved in cellular stress (heat shock protein 70-kDa, HSP70, endoplasmic reticulum (ER stress (immunoglobulin heavy chain binding protein, Bip; proteasome subunit β5, PSMB5 and apoptosis (BCL-2 associated X protein, Bax; cysteine aspartate protease-3, Caspase-3 in bovine blastocysts produced by IVF or SCNT and compare it with that of their in vivo counterparts. Poly (A + mRNA was isolated from three pools of 10 blastocysts per treatment and analyzed by real-time RT-PCR. The RA of three of the stress indicators analyzed (Bax, PSMB5 and Bip was significantly increased in SCNT embryos as compared with that of in vivo-derived blastocysts. No significant differences were found in the RA of HSP70 and Caspase-3 gene transcripts. This study could potentially complement morphological analyses in the development of an effective and accurate technique for the diagnosis of embryo quality, ultimately aiding to improve the efficiency of assisted reproductive techniques (ART.

  14. Test-day somatic cell score, fat-to-protein ratio and milk yield as indicator traits for sub-clinical mastitis in dairy cattle.

    Science.gov (United States)

    Jamrozik, J; Schaeffer, L R

    2012-02-01

    Test-day (TD) records of milk, fat-to-protein ratio (F:P) and somatic cell score (SCS) of first-lactation Canadian Holstein cows were analysed by a three-trait finite mixture random regression model, with the purpose of revealing hidden structures in the data owing to putative, sub-clinical mastitis. Different distributions of the data were allowed in 30 intervals of days in milk (DIM), covering the lactation from 5 to 305 days. Bayesian analysis with Gibbs sampling was used for model inferences. Estimated proportion of TD records originated from cows infected with mastitis was 0.66 in DIM from 5 to 15 and averaged 0.2 in the remaining part of lactation. Data from healthy and mastitic cows exhibited markedly different distributions, with respect to both average value and the variance, across all parts of lactation. Heterogeneity of distributions for infected cows was also apparent in different DIM intervals. Cows with mastitis were characterized by smaller milk yield (down to -5 kg) and larger F:P (up to 0.13) and SCS (up to 1.3) compared with healthy contemporaries. Differences in averages between healthy and infected cows for F:P were the most profound at the beginning of lactation, when a dairy cow suffers the strongest energy deficit and is therefore more prone to mammary infection. Residual variances for data from infected cows were substantially larger than for the other mixture components. Fat-to-protein ratio had a significant genetic component, with estimates of heritability that were larger or comparable with milk yield, and was not strongly correlated with milk and SCS on both genetic and environmental scales. Daily milk, F:P and SCS are easily available from milk-recording data for most breeding schemes in dairy cattle. Fat-to-protein ratio can potentially be a valuable addition to SCS and milk yield as an indicator trait for selection against mastitis.

  15. Co-expression of tenascin-C and vimentin in human breast cancer cells indicates phenotypic transdifferentiation during tumour progression: correlation with histopathological parameters, hormone receptors, and oncoproteins.

    Science.gov (United States)

    Dandachi, N; Hauser-Kronberger, C; Moré, E; Wiesener, B; Hacker, G W; Dietze, O; Wirl, G

    2001-02-01

    Loss of epithelial morphology and the acquisition of mesenchymal characteristics are typical for carcinoma cells in tumour progression. In human breast carcinomas, up-regulation of tenascin-C (TN-C) and vimentin (Vim) is frequently observed in cancer cells and correlates with increased malignancy. Thus, it is possible that TN-C is co-expressed with Vim, representing cancer cells that have undergone epithelial-mesenchymal transition (EMT). This study examined 128 breast carcinomas using immunohistochemical techniques to demonstrate that mammary cancer cells are a prominent source of both TN-C and Vim. Statistical analysis revealed a significant association between TN-C and Vim expression in cancer cells. TN-C expression also correlated positively with overexpression of c-erbB-2 oncoprotein and down-regulation of oestrogen receptors (ERs). Eleven human mammary cancer cell lines and two 'normal' cell lines were examined by western blotting and immunohistochemistry. Co-expression of TN-C and Vim was detected in the carcinosarcoma cell line HS 578T, SK-BR-3 (B), fibroblast-like MDA-MB-231 cells, and the myoepithelial cell line HBL 100. These findings suggest that TN-C and Vim, when co-expressed in mammary carcinoma cells, represent regulator genes likely to be involved in EMT during mammary carcinogenesis.

  16. Investigating Nanosilver Effects on Blood Cells Counter in Male Rats

    Directory of Open Access Journals (Sweden)

    H Aghababa

    2013-03-01

    Full Text Available Introduction: Nanosilver particles are one of the functional grounds in nanotechnology field. These nanoparticles may produce free radicals and destruct different cells. In this study, Nanosilver toxic effects on RBC and WBC numbers in male Rats were studied. Thus, male rats were treated with nanosilver and RBC and WBC were detected. Methods: In this study, RBC was detected in male Wistar rats following exposure to 50, 100, 200 and 400ppm concentration of silver nanoparticles administrated peritoneally. Then, RBC and WBC were collected in rats 3, 8 and 12 days after treatment of nanosilver particles. Numbers of RBC and WBC were compared in treatment and control groups. Results: The study results indicated that dose of 400ppm nanosilver was effective on decrease of RBC and increase of WBC in treatment rats 12 day after treatment. These results were significant (p≤0/01. Discussion: The efficiency of 400ppm concentration of nanosilver, RBC decease and WBC increase could be referred to probabale lyses of RBC cell membranes and sever incitement of cellular immune system. The extra investigation is recommended regarding variety of new shapes, sizes and composition of nanosilver.

  17. Optical tweezers as a new biomedical tool to measure zeta potential of stored red blood cells.

    Directory of Open Access Journals (Sweden)

    Diego C N Silva

    Full Text Available During storage, red blood cells (RBCs for transfusion purposes suffer progressive deterioration. Sialylated glycoproteins of the RBC membrane are responsible for a negatively charged surface which creates a repulsive electrical zeta potential. These charges help prevent the interaction between RBCs and other cells, and especially among each RBCs. Reports in the literature have stated that RBCs sialylated glycoproteins can be sensitive to enzymes released by leukocyte degranulation. Thus, the aim of this study was, by using an optical tweezers as a biomedical tool, to measure the zeta potential in standard RBCs units and in leukocyte reduced RBC units (collected in CPD-SAGM during storage. Optical tweezers is a sensitive tool that uses light for measuring cell biophysical properties which are important for clinical and research purposes. This is the first study to analyze RBCs membrane charges during storage. In addition, we herein also measured the elasticity of RBCs also collected in CPD-SAGM. In conclusion, the zeta potential decreased 42% and cells were 134% less deformable at the end of storage. The zeta potential from leukodepleted units had a similar profile when compared to units stored without leukoreduction, indicating that leukocyte lyses were not responsible for the zeta potential decay. Flow cytometry measurements of reactive oxygen species suggested that this decay is due to membrane oxidative damages. These results show that measurements of zeta potentials provide new insights about RBCs storage lesion for transfusion purposes.

  18. High frequency of cytolytic 21-hydroxylase-specific CD8+ T cells in autoimmune Addison's disease patients.

    Science.gov (United States)

    Dawoodji, Amina; Chen, Ji-Li; Shepherd, Dawn; Dalin, Frida; Tarlton, Andrea; Alimohammadi, Mohammad; Penna-Martinez, Marissa; Meyer, Gesine; Mitchell, Anna L; Gan, Earn H; Bratland, Eirik; Bensing, Sophie; Husebye, Eystein S; Pearce, Simon H; Badenhoop, Klaus; Kämpe, Olle; Cerundolo, Vincenzo

    2014-09-01

    The mechanisms behind destruction of the adrenal glands in autoimmune Addison's disease remain unclear. Autoantibodies against steroid 21-hydroxylase, an intracellular key enzyme of the adrenal cortex, are found in >90% of patients, but these autoantibodies are not thought to mediate the disease. In this article, we demonstrate highly frequent 21-hydroxylase-specific T cells detectable in 20 patients with Addison's disease. Using overlapping 18-aa peptides spanning the full length of 21-hydroxylase, we identified immunodominant CD8(+) and CD4(+) T cell responses in a large proportion of Addison's patients both ex vivo and after in vitro culture of PBLs ≤20 y after diagnosis. In a large proportion of patients, CD8(+) and CD4(+) 21-hydroxylase-specific T cells were very abundant and detectable in ex vivo assays. HLA class I tetramer-guided isolation of 21-hydroxylase-specific CD8(+) T cells showed their ability to lyse 21-hydroxylase-positive target cells, consistent with a potential mechanism for disease pathogenesis. These data indicate that strong CTL responses to 21-hydroxylase often occur in vivo, and that reactive CTLs have substantial proliferative and cytolytic potential. These results have implications for earlier diagnosis of adrenal failure and ultimately a potential target for therapeutic intervention and induction of immunity against adrenal cortex cancer. Copyright © 2014 by The American Association of Immunologists, Inc.

  19. Sucrose-mediated giant cell formation in the genus Neisseria.

    Science.gov (United States)

    Johnson, K G; McDonald, I J

    1976-03-01

    Growth of Neisseria perflava, Neisseria cinerea, and Neisseria sicca strain Kirkland in media supplemented with sucrose (0.5 to 5.0% w/v) resulted in the formation of giant cells. Response to sucrose was specific in that a variety of other carbohydrates did not mediate giant cell formation. Giant cells appeared only under growth conditions and did not lyse upon transfer to medium lacking sucrose or upon resuspension in hypotonic media. Reversion of giant to normal cells occurred when giant cells were used as inocula and allowed to multiply in media lacking sucrose.

  20. Innovative T Cell-Targeted Therapy for Ovarian Cancer

    Science.gov (United States)

    2014-10-01

    A. Tabilio. 2009. Activated autologous T cells exert an anti-B-cell chronic lymphatic leukemia effect in vitro and in vivo. Cytotherapy 11:86-96... leukemia . Sci Transl Med 2011; 3(95): 95ra73. 15. Porter DL, Levine BL, Kalos M, Bagg A, June CH. Chimeric antigen receptor-modified T cells in chronic ...malignancies. Kasumi 3 is a CD33þ CD34þ undifferentiated leukemia cell line that was lysed at intermediate levels by gd T cells. Chronic myelogenous leukemia

  1. Induction of IL21 in Peripheral T Follicular Helper Cells Is an Indicator of Influenza Vaccine Response in a Previously Vaccinated HIV-Infected Pediatric Cohort.

    Science.gov (United States)

    de Armas, Lesley R; Cotugno, Nicola; Pallikkuth, Suresh; Pan, Li; Rinaldi, Stefano; Sanchez, M Celeste; Gonzalez, Louis; Cagigi, Alberto; Rossi, Paolo; Palma, Paolo; Pahwa, Savita

    2017-03-01

    HIV-infected patients of all ages frequently underperform in response to seasonal influenza vaccination, despite virologic control of HIV. The molecular mechanisms governing this impairment, as well as predictive biomarkers for responsiveness, remain unknown. This study was performed in samples obtained prevaccination (T0) from HIV-infected children who received the 2012-2013 seasonal influenza vaccine. Response status was determined based on established criterion for hemagglutination inhibition titer; participants with a hemagglutination titer ≥1:40 plus a ≥4-fold increase over T0 at 3 wk postvaccination were designated as responders. All children had a history of prior influenza vaccinations. At T0, the frequencies of CD4 T cell subsets, including peripheral T follicular helper (pTfh) cells, which provide help to B cells for developing into Ab-secreting cells, were similar between responders and nonresponders. However, in response to in vitro stimulation with influenza A/California/7/2009 (H1N1) Ag, differential gene expression related to pTfh cell function was observed by Fluidigm high-density RT-PCR between responders and nonresponders. In responders, H1N1 stimulation at T0 also resulted in CXCR5 induction (mRNA and protein) in CD4 T cells and IL21 gene induction in pTfh cells that were strongly associated with H1N1-specific B cell responses postvaccination. In contrast, CD4 T cells of nonresponders exhibited increased expression of IL2 and STAT5 genes, which are known to antagonize peripheral Tfh cell function. These results suggest that the quality of pTfh cells at the time of immunization is important for influenza vaccine responses and provide a rationale for targeted, ex vivo Ag-driven molecular profiling of purified immune cells to detect predictive biomarkers of the vaccine response.

  2. Characterization of thermophilic strain Geobacillus sp. SY-9 with capability to lyse bacterial Cells%嗜热溶胞土芽孢杆菌(Geobacillus sp.) SY-9的基本特性

    Institute of Scientific and Technical Information of China (English)

    宋玉栋; 胡洪营; 李鑫

    2007-01-01

    从污泥堆肥中分离得到一株具有溶胞能力的嗜热菌SY-9,经形态及16S rDNA测序初步鉴定为土芽孢杆菌属(Geobacillus sp.).对其生长、产酶及溶胞特性进行了研究.结果表明,SY-9最适生长pH值为7~9,最适生长温度约60℃,60℃的世代时间为34min.SY-9培养上清液具有溶胞能力,上清液经过热处理后溶胞能力明显下降,说明溶胞能力主要来自酶的作用.SY-9间歇培养过程中,培养上清液对大肠杆菌(Escherichia coli)的溶胞活性在SY-9进入稳定生长期后逐渐升高,达到最大值后随着培养时间的延长逐渐下降.SY-9培养上清液对受试的5株革兰氏阴性菌(3株大肠杆菌及2株假单胞菌)及部分革兰氏阳性菌(枯草芽孢杆菌、红球菌、球形节杆菌及溶壁微球菌)都具有溶胞能力.

  3. 改良的碱裂解法提取动物细胞附加体DNA%A modified alkaline lyses protocol for extracting episomal DNA from the mammalian cells

    Institute of Scientific and Technical Information of China (English)

    周海胜; 汪渊; 秦宜德; 储斌; 张胜权; 周青; 沈继龙

    2009-01-01

    目的 利用一种快速、简便的方法 提取动物细胞附加体DNA.方法 通过电转移的方法介导pEGFP-N1转染小鼠成纤维细胞NIH3T3和中国仓鼠卵巢癌细胞CHO等细胞系.转染后48 h,收集转染细胞.用STET缓冲液重悬细胞,加入碱裂解液裂解细胞,再用7.5 mol/L醋酸胺中和.离心收集上清,用1∶1酚/氯仿抽提2次,氯仿抽提1次,乙醇沉淀附加体DNA.TE溶解DNA.提取的DNA通过PCR和southern blot方法进行分析.结果 PCR和Southem blot结果证实利用改良的碱裂解法可以从动物细胞提取附加体DNA.结论 利用简便快速的碱裂解法可以提取哺乳动物细胞附加体DNA.

  4. Human immunodeficiency virus type 1 RNA detection in peripheral blood mononuclear cells by polymerase chain reaction: enhanced sensitivity after mitogenic stimulation.

    Science.gov (United States)

    Tetali, S K; Oyaizu, N; Paul, M; Pahwa, S

    1993-01-01

    The aim of this study was to investigate whether stimulus-induced up-regulation of human immunodeficiency virus type 1 (HIV-1) expression in peripheral blood mononuclear cells (PBMC) could enhance the diagnostic sensitivity of the polymerase chain reaction (PCR). PBMC derived from 11 HIV-1-infected asymptomatic adults were cultured with a stimulus of phytohemagglutinin (PHA) plus phorbol 12-myristate 13-acetate (PMA) for 36 h prior to lysing the cells for PCR. In all 11 patients studied, the intensity of PCR-assisted HIV RNA amplification (RNA-PCR) performed on stimulated cells was significantly (p < 0.001) higher than that obtained on unstimulated cells. A comparison of conventional PCR-assisted DNA amplification (DNA-PCR) with that of RNA-PCR was made on seven patients. The sensitivity of DNA-PCR was also increased by prior stimulation of cells, although not to the same extent as was observed for RNA-PCR. The results of our study indicate that the sensitivity of PCR can be significantly enhanced by prior activation of cells with PHA and PMA.

  5. Selective recruitment of Th I cells induced by re-infection of succeptible and resistant mice with Pseudomonas aerugionosa in the lungs indicates protective role of IL-12

    DEFF Research Database (Denmark)

    Moser, C; Jensen, P O; Kobayashi, O;

    2002-01-01

    for improved clearance of bacteria, was observed when compared with singly-infected mice. The improved outcome in re-infected mice correlated with changes in CD4 cell numbers. Surface expression of LFA-1 on pulmonary CD4 cells was increased in re-infected compared with singly-infected mice. Moreover...

  6. Morphometric evaluation of seminiferous tubule and proportionate numerical analysis of Sertoli and spermatogenic cells indicate differences between crossbred and purebred bulls

    Directory of Open Access Journals (Sweden)

    Utkarsh K. Tripathi

    2015-05-01

    Full Text Available Aim: The present study compared the testicular cytology and histology between crossbred (Holstein–Friesian [HF] × Tharparkar and purebred (HF and Tharparkar bulls to find out differences if any. Materials and Methods: Four peripubertal bulls from each breed were utilized for the study. Through percutaneous needle aspiration biopsy, Sertoli and spermatogenic cells were extracted, and morphometry was studied. For histological studies, testicular tissues obtained through unilateral castration were utilized. Sertoli cells specific GATA4 antibody was used to study the population of Sertoli cells in the seminiferous tubule through immunofluorescence. Results: The testicular weight, volume, and scrotal circumference differed significantly among the breeds. The diameter and area of the seminiferous tubule was high in HF, followed by Karan Fries (KF, and Tharparkar bulls. However, the degree of compactness, based on qualitative evaluation, was high in Tharparkar followed by KF and HF bulls. The intensity of Leydig cells was higher in Tharparkar bulls followed by KF and HF. The proportion of Sertoli cells was higher (p<0.05 in HF and Tharparkar bulls compared to KF bulls. Conclusion: It may be concluded that variations exist in testicular components of the breeds studied and the proportion of Sertoli cells in relation to spermatogenic cells was significantly lower in crossbred bulls compared to purebred bulls.

  7. Heat shock protein 27 is a potential indicator for response to YangZheng XiaoJi and chemotherapy agents in cancer cells.

    Science.gov (United States)

    Owen, Sioned; Zhao, Huishan; Dart, Alwyn; Wang, Yamei; Ruge, Fiona; Gao, Yong; Wei, Cong; Wu, Yiling; Jiang, Wen G

    2016-11-01

    Heat shock protein 27 (HSP27) is a member of the heat shock protein family which has been linked to tumour progression and, most interestingly, to chemotherapy resistance in cancer patients. The present study examined the potential interplay between HSP27 and YangZheng XiaoJi, a traditional Chinese medicine used in cancer treatment. A range of cell lines from different tumour types including pancreatic, lung, gastric, colorectal, breast, prostate and ovarian cancer (both wild-type and resistant) were used. Levels and activation of HSP27 and its potential associated signalling pathways were evaluated by protein array and western blotting. Knockdown of HSP27 in cancer cells was achieved using siRNA. Localisation and co-localisation of HSP27 and other proteins were carried out by immunofluorescence. Cell growth and migration were evaluated in their response to a range of chemotherapeutic agents. The present study first identified, by way of protein array, that YangZheng XiaoJi was able to inhibit the phosphorylation of HSP27 protein in cancer cells. We further demonstrated that HSP27, which is co-localised with caspase-9, can be blocked from localising in focal adhesions and co-localising with caspase-9 by YangZheng XiaoJi. The study also demonstrated that YangZheng XiaoJi was able to sensitise cancer cells including those cells that were resistant to chemotherapy, to chemotherapeutic agents. Finally, knocking down HSP27 markedly reduced the migration of cancer cells and increased the sensitivity of cancer cells to the inhibitory effect on cellular migration by YangZheng XiaoJi. YangZheng XiaoJi can act as an agent in first sensitising cancer cells to chemotherapy and secondly to overcome, to some degree, chemoresistance when used in an appropriate fashion in patients who have active HSP27.

  8. Heat shock protein 27 is a potential indicator for response to YangZheng XiaoJi and chemotherapy agents in cancer cells

    Science.gov (United States)

    Owen, Sioned; Zhao, Huishan; Dart, Alwyn; Wang, Yamei; Ruge, Fiona; Gao, Yong; Wei, Cong; Wu, Yiling; Jiang, Wen G.

    2016-01-01

    Heat shock protein 27 (HSP27) is a member of the heat shock protein family which has been linked to tumour progression and, most interestingly, to chemotherapy resistance in cancer patients. The present study examined the potential interplay between HSP27 and YangZheng XiaoJi, a traditional Chinese medicine used in cancer treatment. A range of cell lines from different tumour types including pancreatic, lung, gastric, colorectal, breast, prostate and ovarian cancer (both wild-type and resistant) were used. Levels and activation of HSP27 and its potential associated signalling pathways were evaluated by protein array and western blotting. Knockdown of HSP27 in cancer cells was achieved using siRNA. Localisation and co-localisation of HSP27 and other proteins were carried out by immunofluorescence. Cell growth and migration were evaluated in their response to a range of chemotherapeutic agents. The present study first identified, by way of protein array, that YangZheng XiaoJi was able to inhibit the phosphorylation of HSP27 protein in cancer cells. We further demonstrated that HSP27, which is co-localised with caspase-9, can be blocked from localising in focal adhesions and co-localising with caspase-9 by YangZheng XiaoJi. The study also demonstrated that YangZheng XiaoJi was able to sensitise cancer cells including those cells that were resistant to chemotherapy, to chemotherapeutic agents. Finally, knocking down HSP27 markedly reduced the migration of cancer cells and increased the sensitivity of cancer cells to the inhibitory effect on cellular migration by YangZheng XiaoJi. YangZheng XiaoJi can act as an agent in first sensitising cancer cells to chemotherapy and secondly to overcome, to some degree, chemoresistance when used in an appropriate fashion in patients who have active HSP27. PMID:27600495

  9. Analysis of Mycobacterium ulcerans-specific T-cell cytokines for diagnosis of Buruli ulcer disease and as potential indicator for disease progression

    Science.gov (United States)

    Antwi-Berko, Daniel; Mubarik, Yusif; Abass, Kabiru Mohammed; Owusu, Wellington; Owusu-Dabo, Ellis; Debrah, Linda Batsa; Debrah, Alexander Yaw; Jacobsen, Marc; Phillips, Richard O.

    2017-01-01

    Background Buruli ulcer disease (BUD), caused by Mycobacterium (M.) ulcerans, is the third most common mycobacterial disease after tuberculosis and leprosy. BUD causes necrotic skin lesions and is a significant problem for health care in the affected countries. As for other mycobacterial infections, T cell mediated immune responses are important for protection and recovery during treatment, but detailed studies investigating these immune responses in BUD patients are scarce. In this study, we aimed to characterise M. ulcerans-specific CD4+ T cell responses in BUD patients and to analyse specific cytokine-producing T cells in the context of disease severity and progression. Methodology/Principal findings For this case-control study, whole blood samples of BUD patients (N = 36, 1.5–17 years of age) and healthy contacts (N = 22, 3–15 years of age) were stimulated with antigen prepared from M. ulcerans and CD4+ T cells were analysed for the expression of TNFα, IFNγ and CD40L by flow cytometry. The proportions and profile of cytokine producing CD4+ T cells was compared between the two study groups and correlated with disease progression and severity. Proportions of cytokine double-positive IFNγ+TNFα+, TNFα+CD40L+, IFNγ+CD40L+ (p = 0.014, p = 0.010, p = 0.002, respectively) and triple positive IFNγ+TNFα+CD40L+ (p = 0.010) producing CD4+ T cell subsets were increased in BUD patients. In addition, TNFα+CD40L-IFNγ- CD4+ T cells differed between patients and controls (p = 0.034). TNFα+CD40L-IFNγ- CD4+ T cells were correlated with lesion size (p = 0.010) and proportion were higher in ‘slow’ healers compared to ‘fast healers’ (p = 0.030). Conclusions We were able to identify M. ulcerans-specific CD4+ T cell subsets with specific cytokine profiles. In particular a CD4+ T cell subset, producing TNFα but not IFNγ and CD40L, showed association with lesion size and healing progress. Further studies are required to investigate, if the identified CD4+ T cell

  10. Genome-wide effects of MELK-inhibitor in triple-negative breast cancer cells indicate context-dependent response with p53 as a key determinant.

    Science.gov (United States)

    Simon, Marisa; Mesmar, Fahmi; Helguero, Luisa; Williams, Cecilia

    2017-01-01

    Triple-negative breast cancer (TNBC) is an aggressive, highly recurrent breast cancer subtype, affecting approximately one-fifth of all breast cancer patients. Subpopulations of treatment-resistant cancer stem cells within the tumors are considered to contribute to disease recurrence. A potential druggable target for such cells is the maternal embryonic leucine-zipper kinase (MELK). MELK expression is upregulated in mammary stem cells and in undifferentiated cancers, where it correlates with poor prognosis and potentially mediates treatment resistance. Several MELK inhibitors have been developed, of which one, OTSSP167, is currently in clinical trials. In order to better understand how MELK and its inhibition influence TNBC, we verified its anti-proliferative and apoptotic effects in claudin-low TNBC cell lines MDA-MB-231 and SUM-159 using MTS assays and/or trypan blue viability assays together with analysis of PARP cleavage. Then, using microarrays, we explored which genes were affected by OTSSP167. We demonstrate that different sets of genes are regulated in MDA-MB-231 and SUM-159, but in both cell lines genes involved in cell cycle, mitosis and protein metabolism and folding were regulated. We identified p53 (TP53) as a potential upstream regulator of the regulated genes. Using western blot we found that OTSSP167 downregulates mutant p53 in all tested TNBC cell lines (MDA-MB-231, SUM-159, and BT-549), but upregulates wild-type p53 in the luminal A subtype MCF-7 cell line. We propose that OTSSP167 might have context-dependent or off-target effects, but that one consistent mechanism of action could involve the destabilization of mutant p53.

  11. Genome-wide effects of MELK-inhibitor in triple-negative breast cancer cells indicate context-dependent response with p53 as a key determinant

    Science.gov (United States)

    Simon, Marisa; Mesmar, Fahmi; Helguero, Luisa

    2017-01-01

    Triple-negative breast cancer (TNBC) is an aggressive, highly recurrent breast cancer subtype, affecting approximately one-fifth of all breast cancer patients. Subpopulations of treatment-resistant cancer stem cells within the tumors are considered to contribute to disease recurrence. A potential druggable target for such cells is the maternal embryonic leucine-zipper kinase (MELK). MELK expression is upregulated in mammary stem cells and in undifferentiated cancers, where it correlates with poor prognosis and potentially mediates treatment resistance. Several MELK inhibitors have been developed, of which one, OTSSP167, is currently in clinical trials. In order to better understand how MELK and its inhibition influence TNBC, we verified its anti-proliferative and apoptotic effects in claudin-low TNBC cell lines MDA-MB-231 and SUM-159 using MTS assays and/or trypan blue viability assays together with analysis of PARP cleavage. Then, using microarrays, we explored which genes were affected by OTSSP167. We demonstrate that different sets of genes are regulated in MDA-MB-231 and SUM-159, but in both cell lines genes involved in cell cycle, mitosis and protein metabolism and folding were regulated. We identified p53 (TP53) as a potential upstream regulator of the regulated genes. Using western blot we found that OTSSP167 downregulates mutant p53 in all tested TNBC cell lines (MDA-MB-231, SUM-159, and BT-549), but upregulates wild-type p53 in the luminal A subtype MCF-7 cell line. We propose that OTSSP167 might have context-dependent or off-target effects, but that one consistent mechanism of action could involve the destabilization of mutant p53. PMID:28235006

  12. Recovery of Epstein--Barr virus from nonproducer neonatal human lymphoid cell transformants. [X radiation

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, G.; Miller, G.

    1979-06-01

    Lymphoid cell lines (LCL) were established by infection of two batches of human umbilical cord lymphocytes with low multiplicities of the B95-8 strain of Epstein--Barr virus. Three of the 17 lines released minute mounts of transforming virus. The rest did not, nor did they make capsid antigen. However virus could be regularly recovered by lethal x-irradiation of transformed cells followed by cocultivation with primary human umbilical cord leukocytes. By this technique transforming activity could be identified in 15 of the 17 lines. These data indicate that these nonproducer human neonatal cell transformants established by EBV infection in vitro possess sufficient genetic information to code for production of biologically active mature virions. X rays alone failed to cause a detectable increase in the number of cells with capsid antigen or to enhance extracellular virus production. EBV-positive human serum blocked rescue if it was added during the first 2 to 4 hr after cocultivation, but not thereafter. Transforming virus could be recovered from x-rayed cells which were immediately thereafter lysed by freezing and thawing. These results suggest that recovery of virus following x-ray and cocultivation is not due to activation of the intracellular virus genome. Rather, it is likely that the method detects small numbers of virions which are cell associated. While transforming virus could regularly be rescued from lymphoblastoid cell lines resulting from in vitro transformation, attempts to rescue virus from Raji or EBV-converted BJAB cells were unsuccessful. This discrepancy suggests differences in genome complexity or in genome-cell interactions in different types of EBV-transformed cells.

  13. Higher percentage of in vitro apoptotic cells at time of diagnosis in patients with chronic lymphocytic leukemia indicate earlier treatment requirement: Ten years follow up

    Directory of Open Access Journals (Sweden)

    Kravić-Stevović Tamara

    2014-01-01

    Full Text Available Introduction. Chronic lymphocytic leukemia (CLL has an extremely variable clinical course. Biological reasons for that wide variation in clinical course and survival rates in CLL patients are not fully understood. Objective. The aim of the study was to evaluate the value of spontaneous apoptosis of CLL cells in vitro determined at presentation of disease, in prediction of treatment requirements and evolution of the CLL. Methods. Malignant B cells were isolated from the whole blood of 30 newly diagnosed CLL patients and cultured for 24 hours in RPMI-1640 medium supplemented with 10% of serum obtained from the same CLL patient. Cells were later fixed and processed for embedding in Epon, or cell smears were prepared and stained with TUNEL technique. Results. Ten-year follow-up revealed that patients with lower percentage of cells in apoptosis at presentation of disease had significant longer time treatment initiation (log rank test p0.05. Conclusion. The results of this study emphasize the importance of apoptosis of CLL cells at the time of the initial diagnosis in pathobiology of this disease. [Projekat Ministarstva nauke Republike Srbije, br. 41025

  14. Genome wide transcriptional analysis of resting and IL2 activated human natural killer cells: gene expression signatures indicative of novel molecular signaling pathways

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    Schmitz Alexander

    2007-07-01

    Full Text Available Abstract Background Human natural killer (NK cells are the key contributors of innate immune response and the effector functions of these cells are enhanced by cytokines such as interleukine 2 (IL2. We utilized genome-wide transcriptional profiling to identify gene expression signatures and pathways in resting and IL2 activated NK cell isolated from peripheral blood of healthy donors. Results Gene expression profiling of resting NK cells showed high expression of a number of cytotoxic factors, cytokines, chemokines and inhibitory and activating surface NK receptors. Resting NK cells expressed many genes associated with cellular quiescence and also appeared to have an active TGFβ (TGFB1 signaling pathway. IL2 stimulation induced rapid downregulation of quiescence associated genes and upregulation of genes associated with cell cycle progression and proliferation. Numerous genes that may enhance immune function and responsiveness including activating receptors (DNAM1, KLRC1 and KLRC3, death receptor ligand (TNFSF6 (FASL and TRAIL, chemokine receptors (CX3CR1, CCR5 and CCR7, interleukin receptors (IL2RG, IL18RAB and IL27RA and members of secretory pathways (DEGS1, FKBP11, SSR3, SEC61G and SLC3A2 were upregulated. The expression profile suggested PI3K/AKT activation and NF-κB activation through multiple pathways (TLR/IL1R, TNF receptor induced and TCR-like possibly involving BCL10. Activation of NFAT signaling was supported by increased expression of many pathway members and downstream target genes. The transcription factor GATA3 was expressed in resting cells while T-BET was upregulated on activation concurrent with the change in cytokine expression profile. The importance of NK cells in innate immune response was also reflected by late increased expression of inflammatory chemotactic factors and receptors and molecules involved in adhesion and lymphocyte trafficking or migration. Conclusion This analysis allowed us to identify genes implicated in

  15. Isocitrate lyase localisation in Saccharomyces cerevisiae cells.

    Science.gov (United States)

    Chaves, R S; Herrero, P; Ordiz, I; Angeles del Brio, M; Moreno, F

    1997-10-01

    The isocitrate lyase from Saccharomyces cerevisiae was only located in the cell cytoplasm. This protein was found not to be associated with cell organelles, even under growth conditions that induce peroxisome proliferation. This conclusion is supported by experiments carried out by damaging the protoplast plasma membrane with DEAE-dextran, by differential centrifugation of osmotically lysed protoplast and by using the green fluorescent protein (GFP) of Aequorea victoria as a reporter fusion tag to localise the subcellular compartment to which isocitrate lyase is targeted.

  16. Increased expression of high-mobility group A2: A novel independent indicator of poor prognosis in patients with esophageal squamous cell carcinoma

    Directory of Open Access Journals (Sweden)

    Rongna Wei

    2016-01-01

    Conclusions: High HMGA2 expression was related to lymph node metastasis and poor prognosis in ESCC. Our results indicated that HMGA2 could act as a potential biomarker for prognosis evaluation of ESCC patients.

  17. Local lysis with Alteplase for the treatment of acute embolic leg ischemia following the use of the Duett {sup trademark} closure device: preliminary results; Lokale Alteplase-Lyse zur Therapie der akuten embolischen Beinischaemie nach Einsatz des Duett {sup trademark} -Verschluss-Systems: vorlaeufige Ergebnisse

    Energy Technology Data Exchange (ETDEWEB)

    Schuermann, K.; Buecker, A.; Wingen, M.; Tacke, J.; Wein, B.; Guenther, R.W. [Klinik fuer Radiologische Diagnostik des Universitaetsklinikums der RWTH Aachen (Germany); Janssens, U. [Medizinische Klinik I (Kardiologie) des Universitaetsklinikums der RWTH Aachen (Germany)

    2004-04-01

    unter intravaskulaeren Ballonschutz der arteriellen Punktionsstelle in den Punktionskanal gespritzt wird. Bei 9 Patienten (0,64%) wurde das Prokoagulans unbemerkt in die Femoralarterie injiziert, und es trat eine akute Beinischaemie auf. Acht Patienten erhielten ueber einen kontralateralen femoralen Zugang eine lokale Alteplase-Lyse. Ein Patient wurde operiert. Im Mittel wurden 21 mg Alteplase (4-35 mg) in 14 h (4-21 h) appliziert. Der Verlauf der Lyse wurde angiographisch und klinisch kontrolliert. Die Patienten wurden nach 23 Monaten (4-35 Monaten) telefonisch befragt. Ergebnisse: Bei 3 Patienten war die Lyse vollstaendig, bei 5 Patienten verblieb wenig Restmaterial. In allen Faellen bildete sich die Ischaemiesymptomatik vollstaendig innerhalb der ersten Stunden nach Lysebeginn zurueck. In 5 Faellen trat waehrend der Lyse eine Blutung aus der mit dem Duett-System verschlossenen Punktionsstelle auf, in 2 Faellen mit Ausbildung eines falschen Aneurysmas. Dies fuehrte zum vorzeitigen Ende (n=2) oder zur Unterbrechung der Lyse (n=3). Alle Komplikationen wurden konservativ behandelt. Klinisch verblieben als Spaetfolgen bei 2 lysierten und dem operierten Patienten Par- und Hypaesthesien am Unterschenkel bzw. Fuss. (orig.)

  18. Ubiquitin Specific Protease 26 (USP26 expression analysis in human testicular and extragonadal tissues indicates diverse action of USP26 in cell differentiation and tumorigenesis.

    Directory of Open Access Journals (Sweden)

    Matthew S Wosnitzer

    Full Text Available Ubiquitin specific protease 26 (USP26, a deubiquitinating enzyme, is highly expressed early during murine spermatogenesis, in round spermatids, and at the blood-testis barrier. USP26 has also been recognized as a regulator of androgen receptor (AR hormone-induced action involved in spermatogenesis and steroid production in in vitro studies. Prior mutation screening of USP26 demonstrated an association with human male infertility and low testosterone production, but protein localization and expression in the human testis has not been characterized previously. USP26 expression analysis of mRNA and protein was completed using murine and human testis tissue and human tissue arrays. USP26 and AR mRNA levels in human testis were quantitated using multiplex qRT-PCR. Immunofluorescence colocalization studies were performed with formalin-fixed/paraffin-embedded and frozen tissues using primary and secondary antibodies to detect USP26 and AR protein expression. Human microarray dot blots were used to identify protein expression in extra-gonadal tissues. For the first time, expression of USP26 and colocalization of USP26 with androgen receptor in human testis has been confirmed predominantly in Leydig cell nuclei, with less in Leydig cell cytoplasm, spermatogonia, primary spermatocytes, round spermatids, and Sertoli cells. USP26 likely affects regulatory proteins of early spermatogenesis, including androgen receptor with additional activity in round spermatids. This X-linked gene is not testis-specific, with USP26 mRNA and protein expression identified in multiple other human organ tissues (benign and malignant including androgen-dependent tissues such as breast (myoepithelial cells and secretory luminal cells and thyroid tissue (follicular cells. USP26/AR expression and interaction in spermatogenesis and androgen-dependent cancer warrants additional study and may prove useful in diagnosis and management of male infertility.

  19. Evaluation of clinical trial eligibility and prognostic indices in a population-based cohort of systemic peripheral T-cell lymphomas from the Danish Lymphoma Registry

    DEFF Research Database (Denmark)

    Pedersen, Martin Bjerregård; Hamilton-Dutoit, Stephen Jacques; Bendix, Knud

    2014-01-01

    . Overall, 41% of the patients were considered eligible for interventional CTs implicating curatively intended multiagent chemotherapy, including, if considered appropriate, consolidating stem cell transplantation (SCT), as part of the upfront management strategy. Moreover, 28% was elderly fit and eligible...... for interventional CT, including those with SCT as part of the trial design. Approximately 7% were defined as 'too frail' for aggressive treatment schedules, whereas 24% were deemed not to be eligible for any CT. Both overall and progression-free survivals were effectively predicted by IPI and PIT (p...-positive anaplastic large cell lymphoma patients were significantly younger (median age 40 vs. 62, pnegative counterparts (p

  20. SAP domain-dependent Mkl1 signaling stimulates proliferation and cell migration by induction of a distinct gene set indicative of poor prognosis in breast cancer patients

    Science.gov (United States)

    2014-01-01

    Background The main cause of death of breast cancer patients is not the primary tumor itself but the metastatic disease. Identifying breast cancer-specific signatures for metastasis and learning more about the nature of the genes involved in the metastatic process would 1) improve our understanding of the mechanisms of cancer progression and 2) reveal new therapeutic targets. Previous studies showed that the transcriptional regulator megakaryoblastic leukemia-1 (Mkl1) induces tenascin-C expression in normal and transformed mammary epithelial cells. Tenascin-C is known to be expressed in metastatic niches, is highly induced in cancer stroma and promotes breast cancer metastasis to the lung. Methods Using HC11 mammary epithelial cells overexpressing different Mkl1 constructs, we devised a subtractive transcript profiling screen to identify the mechanism by which Mkl1 induces a gene set co-regulated with tenascin-C. We performed computational analysis of the Mkl1 target genes and used cell biological experiments to confirm the effect of these gene products on cell behavior. To analyze whether this gene set is prognostic of accelerated cancer progression in human patients, we used the bioinformatics tool GOBO that allowed us to investigate a large breast tumor data set linked to patient data. Results We discovered a breast cancer-specific set of genes including tenascin-C, which is regulated by Mkl1 in a SAP domain-dependent, serum response factor-independent manner and is strongly implicated in cell proliferation, cell motility and cancer. Downregulation of this set of transcripts by overexpression of Mkl1 lacking the SAP domain inhibited cell growth and cell migration. Many of these genes are direct Mkl1 targets since their promoter-reporter constructs were induced by Mkl1 in a SAP domain-dependent manner. Transcripts, most strongly reduced in the absence of the SAP domain were mechanoresponsive. Finally, expression of this gene set is associated with high

  1. Effects of two copper compounds on Microcystis aeruginosa cell density, membrane integrity, and microcystin release.

    Science.gov (United States)

    Tsai, Kuo-Pei

    2015-10-01

    Microcystin release following Microcystis aeruginosa cell lysis after copper-based algaecide treatment is often cited as a concern leading to restricted use of algaecide in restoration of natural water resources. To examine this concern, bench-scale experiments were conducted to study responses of M. aeruginosa to 8-day copper exposures as copper sulfate and copper-ethanolamine (Cu-EA). M. aeruginosa UTEX 2385 was cultured in BG11 medium to cell density of 10(6)cells/mL with total and extracellular microcystin of 93 and 53μg/L, respectively. Exposures of copper concentration ranged from 40 to 1000μgCu/L. Cell membrane integrity was indicated by erythrosine B. In the end of experiment, total microcystin and cell density in untreated control (313μg/L and 10(7)cells/mL) was 3.3 and 10 times greater than pretreatment value, respectively. Minimum amount of copper required to reduce M. aeruginosa population within 8 days was 160μgCu/L as copper sulfate and 80μgCu/L as Cu-EA, where total and extracellular microcystin concentrations (47 and 44μg/L for copper sulfate; 56 and 44μg/L for Cu-EA) were degraded with degradation rate coefficient 0.1 day(-1) and were less than pretreatment values. Given a copper concentration at 80µgCu/L as Cu-EA, M. aeruginosa cells were intact and less microcystin were released compared to treatments at 160-1000µgCu/L, where lysed cells and relatively greater microcystin release were observed. Based on the laboratory results, a minimum amount of copper required for reducing M. aeruginosa population could decrease total microcystin concentration and not compromise cells and minimize microcystin release.

  2. Tungsten effects on phosphate-dependent biochemical pathways are species and liver cell line dependent.

    Science.gov (United States)

    Johnson, David R; Ang, Chooyaw; Bednar, Anthony J; Inouye, Laura S

    2010-08-01

    Tungsten, in the form of tungstate, polymerizes with phosphate, and as extensive polymerization occurs, cellular phosphorylation and dephosphorylation reactions may be disrupted, resulting in negative effects on cellular functions. A series of studies were conducted to evaluate the effect of tungsten on several phosphate-dependent intracellular functions, including energy cycling (ATP), regulation of enzyme activity (cytosolic protein tyrosine kinase [cytPTK] and tyrosine phosphatase), and intracellular secondary messengers (cyclic adenosine monophosphate [cAMP]). Rat noncancerous hepatocyte (Clone-9), rat cancerous hepatocyte (H4IIE), and human cancerous hepatocyte (HepG2) cells were exposed to 1-1000 mg/l tungsten (in the form of sodium tungstate) for 24 h, lysed, and analyzed for the above biochemical parameters. Cellular ATP levels were not significantly affected in any cell line. After 4 h, tungsten significantly decreased cytPTK activity in Clone-9 cells at >or= 18 mg/l, had no effect in H4IIE cells, and significantly increased cytPTK activity by 70% in HepG2 cells at >or= 2 mg/l. CytPTK displayed a slight hormetic response to tungsten after 24-h exposure yet returned to normal after 48-h exposure. Tungsten significantly increased cAMP by over 60% in Clone-9 cells at >or= 100 mg/l, significantly increased cAMP in H4IIE cells at only 100 mg/l, and significantly increased cAMP in HepG2 cells between 1-100 mg/l but at much more modest levels (8-20%). In conclusion, these data indicate that tungsten produces complex results that must be carefully interpreted in the context of their respective animal models, as well as the phenotype of the cell lines (i.e., normal vs. cancerous).

  3. Montelukast Disposition: No Indication of Transporter-Mediated Uptake in OATP2B1 and OATP1B1 Expressing HEK293 Cells

    Science.gov (United States)

    Brännström, Marie; Nordell, Pär; Bonn, Britta; Davis, Andrew M.; Palmgren, Anna-Pia; Hilgendorf, Constanze; Rubin, Katarina; Grime, Ken

    2015-01-01

    Clinical studies with montelukast show variability in effect and polymorphic OATP2B1-dependent absorption has previously been implicated as a possible cause. This claim has been challenged with conflicting data and here we used OATP2B1-transfected HEK293 cells to clarify the mechanisms involved. For montelukast, no significant difference in cell uptake between HEK-OATP2B1 and empty vector cell lines was observed at pH 6.5 or pH 7.4, and no concentration-dependent uptake was detected. Montelukast is a carboxylic acid, a relatively potent inhibitor of OATP1B1, OATP1B3, and OATP2B1, and has previously been postulated to be actively transported into human hepatocytes. Using OATP1B1-transfected HEK293 cells and primary human hepatocytes in the presence of OATP inhibitors we demonstrate for the first time that active OATP-dependent transport is unlikely to play a significant role in the human disposition of montelukast. PMID:26694455

  4. Oropharyngeal malignant epithelial cell, lymphocyte and macrophage CD44 surface receptors for hyaluronate are expressed in sustained EBV infection: immunohistochemical data and EBV DNA tissue indices.

    Science.gov (United States)

    Groma, Valerija; Kazanceva, Anna; Nora-Krukle, Zaiga; Murovska, Modra

    2012-09-15

    The role of CD44 in Epstein-Barr virus (EBV)-related epithelial tumors is poorly understood. We studied the expression of CD44 in EBV infection in patients with oral squamous cell carcinoma (SCC) and nasopharyngeal carcinoma (NPC) and measured the EBV DNA. Whole blood, plasma and tissue samples from 8 male and 2 female patients with oral SCC, NPC, salivary gland lymphoepithelioma, normal salivary gland and buccal mucosa were assayed for EBV DNA. Expression of CD44, latent membrane protein (LMP), and labeling of lymphocytes, macrophages and dendritic cells were estimated by immunohistochemistry. Tissue EBV DNA was detected in 7 of 8 cases (87.5%) of oral malignant, benign and border-line lesions. LMP expression levels in tumors varied from absence and minimal to moderate - 50.3, 43.6, 6.0% and 91.1, 6.7, 2.2% for SCC and NPC, respectively. Levels of CD44 positivity in neoplasms were minimal (15.5 and 16.7%), moderate (30.3 and 47.8%), and diffuse (54.2 and 35.5%) for SCC and NPC, respectively, thus deviating from normal oral mucosa revealing heavily stained (100.0%) epithelial contours. CD19-positive B lymphocytes and S100-positive dendritic cells were intermixed with neoplastic cells. Collectively, CD44 mediated signaling may be implicated in EBV infection associated with the pathogenesis of oral SCC and NPC.

  5. Association of cancer metabolism-related proteins with oral carcinogenesis - indications for chemoprevention and metabolic sensitizing of oral squamous cell carcinoma?

    Science.gov (United States)

    Grimm, Martin; Cetindis, Marcel; Lehmann, Max; Biegner, Thorsten; Munz, Adelheid; Teriete, Peter; Kraut, Wiebke; Reinert, Siegmar

    2014-07-21

    Tumor metabolism is a crucial factor for the carcinogenesis of oral squamous cell carcinoma (OSCC). Expression of IGF-R1, glycolysis-related proteins (GLUT-1, HK 2, PFK-1, LDHA, TKTL1), mitochondrial enzymes (SDHA, SDHB, ATP synthase) were analyzed in normal oral mucosa (n = 5), oral precursor lesions (simple hyperplasia, n = 11; squamous intraepithelial neoplasia, SIN I-III, n = 35), and OSCC specimen (n = 42) by immunohistochemistry and real-time polymerase chain reaction (qPCR) analysis in OSCC cell lines. Metabolism-related proteins were correlated with proliferation activity (Ki-67) and apoptotic properties (TUNEL assay) in OSCC. Specificity of antibodies was confirmed by western blotting in cancer cell lines. Expression of IGF-R1, glycolysis-related proteins (GLUT-1, HK 2, LDHA, TKTL1), and mitochondrial enzymes (SDHA, SDHB, ATP synthase) were significantly increased in the carcinogenesis of OSCC. Metabolic active regions of OSCC were strongly correlated with proliferating cancer (Ki-67+) cells without detection of apoptosis (TUNEL assay). This study provides the first evidence of the expression of IGF-R1, glycolysis-related proteins GLUT-1, HK 2, PFK-1, LDHA, and TKTL1, as well as mitochondrial enzymes SDHA, SDHB, and ATP synthase in the multi-step carcinogenesis of OSCC. Both, hypoxia-related glucose metabolism and mitochondrial oxidative phosphorylation characteristics are associated with the carcinogenesis of OSCC. Acidosis and OXPHOS may drive a metabolic shift towards the pentose phosphate pathway (PPP). Therefore, inhibition of the PPP, glycolysis, and targeted anti-mitochondrial therapies (ROS generation) by natural compounds or synthetic vitamin derivatives may act as sensitizer for apoptosis in cancer cells mediated by adjuvant therapies in OSCC.

  6. Maternal plasma levels of cell-free β-HCG mRNA as a prenatal diagnostic indicator of placenta accrete.

    Science.gov (United States)

    Zhou, J; Li, J; Yan, P; Ye, Y H; Peng, W; Wang, S; Wang, X Tong

    2014-09-01

    Several biomarkers, including maternal serum creatinine kinase and α-fetoprotein, have been described as potential tools for the diagnosis of placental abnormalities. This study aimed to determine whether maternal plasma mRNA levels of the β subunit of human chorionic gonadotropin (β-HCG) could predict placenta accreta prenatally. Sixty-eight singleton pregnant women with prior cesarean deliveries (CDs) were classified into three groups: normal placentation (35 women, control group); placenta previa alone (21 women, placenta previa group); and both placenta previa and placenta accreta (12 women, placenta previa/accreta group). Maternal plasma concentrations of cell-free β-HCG mRNA were measured by real-time reverse-transcription polymerase chain reaction and were expressed as multiples of the median (MoM). Cell-free β-HCG mRNA concentrations (MoM, range) were significantly higher in women with placenta accreta (3.65, 2.78-7.19) than in women with placenta previa (0.94, 0.00-2.97) or normal placentation (1.00, 0.00-2.69) (Steel-Dwass test, P placenta previa/accreta group, the concentration of cell-free β-HCG mRNA was significantly higher among women who underwent CDs with hysterectomy (4.41, 3.49-7.19) than among women whose CDs did not result in hysterectomy (3.20, 2.78-3.70) (Mann-Whitney U test, P = 0.012). An increased level of cell-free β-HCG mRNA in the maternal plasma of women with placenta accreta may arise from direct uteroplacental transfer of cell-free placental mRNA molecules. The concentration of cell-free β-HCG mRNA in maternal plasma may be applicable to the prenatal diagnosis of placenta accreta, especially to identify women with placenta accreta likely to require hysterectomy. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. A study on body mass index, blood pressure, and red blood cell indices in new entering students of the University of Isfahan

    Directory of Open Access Journals (Sweden)

    Alireza Moafi

    2011-01-01

    Conclusions: High incidence of abnormal BMI in the study population, and its association with systolic blood pressure indicate the importance of nutritional guidelines and counseling programs for freshman students. On the other hand, high incidence of anemia in this population ascertains the necessity of anemia screening programs before academic studies.

  8. TO EVALUATE ANAEMIA BY ERYTHROCYTE INDICES, RED CELL DISTRIBUTION WIDTH AND HAEMOGLOBIN ELECTROPHORESIS WITH SPECIAL REFERENCE TO THALASSEMIA IN PAEDIATRIC AGE GROUP

    Directory of Open Access Journals (Sweden)

    Mahendra

    2015-02-01

    Full Text Available Anemia is not a diagnosis; it is a manifestation of an underlying disorder. Thus, even mild, asymptomatic anemia should be investigated so that the primary problem can be diagnosed and treated. Laboratory evaluation begins with a CBC, including WBC and platelet counts, RBC indices and morphology (MCV, MCH, MCHC, RBC distribution width [RDW], and examination of the peripheral smear. In many instances routine test like Hb, TLC, DLC, GBP fail to decide anemia especially in early cases and also fail to decide the type of anaemia. In such situations the RBC indices and RDW are very useful. These become abnormal even before changes in routine hemogram are appreciable. Thalassemia minor poses problems in diagnosis because GBP reveals no features of hemolysis rather it has microcytic hypochromic picture which has similarity with iron deficiency anemia. It is difficult to differentiate between two by only GBP. Several decision making rules have been proposed for differentiation. METHOD : The present study was carried out in 100 cases to evaluate anaemia in different age groups based on RBC Indices and RDW and t o evaluate sensitivity of RBC indices and RDW in diagnosis of anaemia. Cases showing positivity by various rules and RDW in favour of thalassemia minor were subjected to Hb electrophoresis for confirmation of diagnosis. RESULTS : RBC indices are more sensitive for diagnosis of microcytic hypochromic anemia, normocytic normochromic anemia and macrocytic anemia than PBS alone. RDW - CV is superior to all in use to differentiate iron deficiency anemia an d thalassemia minor having high sensitivity 87.3% and specificity 90 .5%.

  9. CXCR4 Expression in Gastric Cancer and Bone Marrow: Association with Hypoxia-Regulated Indices, Disseminated Tumor Cells, and Patients Survival

    Directory of Open Access Journals (Sweden)

    Dmitry Osinsky

    2015-01-01

    Full Text Available Aim. The analysis of the association of CXCR4 expression in gastric cancer (GC and bone marrow (BM with clinical characteristics. Patients and Methods. 65 patients with GC were investigated. Immunohistochemistry, immunocytochemistry, NMR-spectroscopy, and zymography were used. Results. CXCR4 was expressed in 78.5% of GC specimens and correlated with tumor hypoxia (P<0.05, VEGF expression (P<0.01, and gelatinases activity (P<0.05. CXCR4-positive cells in GC were detected in 80% of patients with disseminated tumor cells (DTCs. Overall survival (OS of patients with CXCR4-positive tumors was poorer than that of patients with CXCR4-negative tumors (P=0.037. The CXCR4-positive cells in BM were found in 46% of all patients and in 56% of patients with DTCs. CXCR4 expression in BM was not associated with OS. Risk of unfavourable outcome is increased in patients with CXCR4-positive tumors (P<0.05. CXCR4 expression in BM was positively associated with DTCs, especially in patients with M0 category. Risk of unfavourable outcome is increased in patients with M0 category and with both CXCR4-positive BM and DTCs (P=0.03. Conclusions. CXCR4 expression in tumor was positively correlated with hypoxia level and VEGF expression in tumor as well as OS. CXCR4 expression in BM is associated with DTCs.

  10. Natural Killer Cells Are Activated by Lactic Acid Bacteria-Matured Dendritic Cells

    DEFF Research Database (Denmark)

    Fink, Lisbeth Nielsen; Christensen, Hanne Risager; Frøkiær, Hanne

    Natural killer (NK) cells are cells of the non-specific immune system lysing altered self-cells. A non-cytolytic subset of NK cells may serve a regulatory role by secreting cytokines. Bacteria translocating across the gastrointestinal mucosa are presumed to gain access to NK cells, as consumption...... of certain lactic acid bacteria has been shown to increase in vivo NK cytotoxicity. Here, we investigated how human gut flora-derived lactobacilli affect NK cells in vitro, by measuring proliferation and IFN-gamma production of human NK cells upon bacterial stimulation. Human peripheral blood NK cells were...... incubated with 10 microg/ml UV-inactivated bacteria or 10 microg/ml phytohemagglutinin (PHA) for four days. Proliferation was assessed by incorporation of radioactive thymidine into NK cell DNA. The IFN-gamma concentration was measured by ELISA. Incubation of NK cells with a Lactobacillus acidophilus strain...

  11. Divergent effects of liraglutide, exendin-4, and sitagliptin on beta-cell mass and indicators of pancreatitis in a mouse model of hyperglycaemia.

    Directory of Open Access Journals (Sweden)

    Angeles Mondragon

    Full Text Available Glucagon-like peptide-1 (GLP-1 receptor agonists and dipeptidyl peptidase-4 (DPP4 inhibitors improve glucose tolerance by still incompletely understood mechanisms. Each class of antihyperglycemic drugs has also been proposed to increase pancreatitis risk. Here, we compare systematically the effects of two widely-used GLP-1 analogues, liraglutide and exendin-4, and the DPP4 inhibitor, sitagliptin, in the mouse.C57BL6 mice were maintained for 131 days on a normal diet (ND or a diet comprising 60% fat (HFD before measurements of fasting blood glucose and insulin, and intraperitoneal glucose tolerance. Beta- and alpha- cell volume, and Reg3b immunoreactivity, were measured by immunohistochemical analysis of pancreatic slices.Whereas liraglutide (200 µg/kg and exendin-4 (10 µg/kg treatment reduced body weight and/or improved glucose tolerance, sitagliptin (10 mg/kg was without effect on either parameter. Liraglutide caused a sharp reduction in beta-cell mass in both ND and HFD mice, whereas exendin-4 exerted no effect. By contrast, sitagliptin unmasked an action of high fat diet to increase beta-cell mass. Reg3B positive area was augmented by all three agents in normal chow-fed mice, whilst sitagliptin and exendin-4, but not liraglutide, affected this parameter in HFD animals. Correspondingly sitagliptin, but not the GLP-1 analogues, increased circulating amylase levels in ND and HFD mice.Liraglutide improves glucose tolerance in the mouse whilst exerting relatively modest effects on pancreatitis risk. Conversely, exendin-4 and sitagliptin, at doses which exert, respectively, minor or no effects on metabolic parameters, lead to signs of pancreatitis.

  12. P16INK4a Positive Cells in Human Skin Are Indicative of Local Elastic Fiber Morphology, Facial Wrinkling, and Perceived Age

    DEFF Research Database (Denmark)

    Waaijer, Mariëtte E C; Gunn, David A; Adams, Peter D

    2016-01-01

    from 178 participants (aged 45-81 years) of the Leiden Longevity Study. Local elastic fiber morphology, facial wrinkles, and perceived facial age were compared to tertiles of p16INK4a counts, while adjusting for chronological age and other potential confounders. The numbers of epidermal and dermal p16...... wrinkles and a higher perceived age. Participants in the lowest tertile of epidermal p16INK4a counts looked 3 years younger than those in the highest tertile, independently of chronological age and elastic fiber morphology. In conclusion, p16INK4a positive cell numbers in sun-protected human arm skin...

  13. Detection of Human Papillomavirus 16-Specific IgG and IgM Antibodies in Patient Sera: A Potential Indicator of Oral Squamous Cell Carcinoma Risk Factor

    OpenAIRE

    Kerishnan, Jesinda P.; Subash C B Gopinath; Kai, Sia Bik; Tang, Thean-Hock; Ng, Helen Lee-Ching; Rahman, Zainal Ariff Abdul; Hashim, Uda; Chen, Yeng

    2016-01-01

    The association between human papillomavirus type 16 (HPV16) and oral cancer has been widely reported. However, detecting anti-HPV antibodies in patient sera to determine risk for oral squamous cell carcinoma (OSCC) has not been well studied. In the present investigation, a total of 206 OSCC serum samples from the Malaysian Oral Cancer Database & Tissue Bank System, with 134 control serum samples, were analyzed by enzyme-linked immunosorbant assay (ELISA) to detect HPV16-specific IgG and IgM ...

  14. Composition of Herba Pogostemonis water extract and protection of infected mice against Salmonella Typhimurium-induced liver damage and mortality by stimulation of innate immune cells.

    Science.gov (United States)

    Kim, Sung Phil; Moon, Eunpyo; Nam, Seok Hyun; Friedman, Mendel

    2012-12-12

    GC-MS analysis of a hot water extract of Herba Pogostemonis (HP) revealed the presence of 131 compounds. HP slightly inhibited Salmonella Typhimurium bacteria in culture and stimulated uptake of the bacteria into RAW 264.7 murine macrophage cells as indicated by both increased fluorescence from internalized FITC-dextran and increased colony-forming unit (CFU) counts of the lysed macrophages. Postinfection, the HP-treated cells showed lower bacterial counts than the control. HP elicited altered morphology, elevated inducible NO synthase (iNOS) mRNA, and reduced pro-inflammatory cytokine expression in macrophage cells. Salmonella induced increased expression of iNOS mRNA, cognate polypeptides, and NO. Histology of mice infected with a sublethal dose (1 × 10(4) CFU) of Salmonella showed that intraperitoneally administered HP protected against necrosis of the liver, a biomarker of in vivo salmonellosis. The lifespan of mice infected with a lethal dose (1 × 10(5) CFU) was significantly extended. These results suggest that the activity of HP against bacterial infection in mice occurs through the activation of innate immune macrophage cells. The relationship of composition of HP to bioactivity is discussed.

  15. MUC16 provides immune protection by inhibiting synapse formation between NK and ovarian tumor cells

    Directory of Open Access Journals (Sweden)

    Migneault Martine

    2010-01-01

    Full Text Available Abstract Background Cancer cells utilize a variety of mechanisms to evade immune detection and attack. Effective immune detection largely relies on the formation of an immune synapse which requires close contact between immune cells and their targets. Here, we show that MUC16, a heavily glycosylated 3-5 million Da mucin expressed on the surface of ovarian tumor cells, inhibits the formation of immune synapses between NK cells and ovarian tumor targets. Our results indicate that MUC16-mediated inhibition of immune synapse formation is an effective mechanism employed by ovarian tumors to evade immune recognition. Results Expression of low levels of MUC16 strongly correlated with an increased number of conjugates and activating immune synapses between ovarian tumor cells and primary naïve NK cells. MUC16-knockdown ovarian tumor cells were more susceptible to lysis by primary NK cells than MUC16 expressing controls. This increased lysis was not due to differences in the expression levels of the ligands for the activating receptors DNAM-1 and NKG2D. The NK cell leukemia cell line (NKL, which does not express KIRs but are positive for DNAM-1 and NKG2D, also conjugated and lysed MUC16-knockdown cells more efficiently than MUC16 expressing controls. Tumor cells that survived the NKL challenge expressed higher levels of MUC16 indicating selective lysis of MUC16low targets. The higher csMUC16 levels on the NKL resistant tumor cells correlated with more protection from lysis as compared to target cells that were never exposed to the effectors. Conclusion MUC16, a carrier of the tumor marker CA125, has previously been shown to facilitate ovarian tumor metastasis and inhibits NK cell mediated lysis of tumor targets. Our data now demonstrates that MUC16 expressing ovarian cancer cells are protected from recognition by NK cells. The immune protection provided by MUC16 may lead to selective survival of ovarian cancer cells that are more efficient in

  16. MUC16 provides immune protection by inhibiting synapse formation between NK and ovarian tumor cells.

    Science.gov (United States)

    Gubbels, Jennifer A A; Felder, Mildred; Horibata, Sachi; Belisle, Jennifer A; Kapur, Arvinder; Holden, Helen; Petrie, Sarah; Migneault, Martine; Rancourt, Claudine; Connor, Joseph P; Patankar, Manish S

    2010-01-20

    Cancer cells utilize a variety of mechanisms to evade immune detection and attack. Effective immune detection largely relies on the formation of an immune synapse which requires close contact between immune cells and their targets. Here, we show that MUC16, a heavily glycosylated 3-5 million Da mucin expressed on the surface of ovarian tumor cells, inhibits the formation of immune synapses between NK cells and ovarian tumor targets. Our results indicate that MUC16-mediated inhibition of immune synapse formation is an effective mechanism employed by ovarian tumors to evade immune recognition. Expression of low levels of MUC16 strongly correlated with an increased number of conjugates and activating immune synapses between ovarian tumor cells and primary naïve NK cells. MUC16-knockdown ovarian tumor cells were more susceptible to lysis by primary NK cells than MUC16 expressing controls. This increased lysis was not due to differences in the expression levels of the ligands for the activating receptors DNAM-1 and NKG2D. The NK cell leukemia cell line (NKL), which does not express KIRs but are positive for DNAM-1 and NKG2D, also conjugated and lysed MUC16-knockdown cells more efficiently than MUC16 expressing controls. Tumor cells that survived the NKL challenge expressed higher levels of MUC16 indicating selective lysis of MUC16(low) targets. The higher csMUC16 levels on the NKL resistant tumor cells correlated with more protection from lysis as compared to target cells that were never exposed to the effectors. MUC16, a carrier of the tumor marker CA125, has previously been shown to facilitate ovarian tumor metastasis and inhibits NK cell mediated lysis of tumor targets. Our data now demonstrates that MUC16 expressing ovarian cancer cells are protected from recognition by NK cells. The immune protection provided by MUC16 may lead to selective survival of ovarian cancer cells that are more efficient in metastasizing within the peritoneal cavity and also at overcoming

  17. The relevance of serum carcinoembryonic antigen as an indicator of brain metastasis detection in advanced non-small cell lung cancer.

    Science.gov (United States)

    Lee, Dong-Soo; Kim, Yeon-Sil; Jung, So-Lyoung; Lee, Kyo-Young; Kang, Jin-Hyoung; Park, Sarah; Kim, Young-Kyoon; Yoo, Ie-Ryung; Choi, Byung-Ock; Jang, Hong-Seok; Yoon, Sei-Chul

    2012-08-01

    Although many biomarkers have emerged in non-small cell lung cancer (NSCLC), the predictive value of site-specific spread is not fully defined. We designed this study to determine if there is an association between serum biomarkers and brain metastasis in advanced NSCLC. We evaluated 227 eligible advanced NSCLC patients between May 2005 and March 2010. Patients who had been newly diagnosed with stage IV NSCLC but had not received treatment previously, and had available information on at least one of the following pretreatment serum biomarkers were enrolled: carcinoembryonic antigen (CEA), cytokeratin 19 fragments (CYFRA 21-1), cancer antigen 125 (CA 125), cancer antigen 19-9, and squamous cancer cell antigen. Whole body imaging studies and magnetic resonance imaging of the brain were reviewed, and the total number of metastatic regions was scored. Brain metastasis was detected in 66 (29.1%) patients. Although serum CEA, CYFRA 21-1, and CA 125 levels were significantly different between low total metastatic score group (score 1-3) and high total metastatic score group (score 4-7), only CEA level was significantly different between patients with brain metastasis and those without brain metastasis (p present study demonstrated that the pretreatment serum CEA level was significantly correlated with brain metastasis in advanced NSCLC. These findings suggested the possible role of CEA in the pathogenesis of brain invasion. More vigilant surveillance would be warranted in the high-risk group of patients with high serum CEA level and multiple synchronous metastasis.

  18. In vitro generation of human cytotoxic lymphocytes by virus. Viral glycoproteins induce nonspecific cell-mediated cytotoxicity without release of interferon

    OpenAIRE

    1981-01-01

    Purified hemagglutinin and fusion glycoproteins of measles virus either in soluble form or inserted in artifical membranes bind to human peripheral blood lymphocytes and induce cell-mediated cytotoxicity (CMC) in a dose-response fashion. Both autologous and heterologous noninfected target cells are lysed in vitro. The expression of CMC is not inhibited by anti-measles virus antibody added to lymphocytes previously exposed to viral glycoproteins. THe killer lymphocytes are Fc receptor positive...

  19. Differential expression of inwardly rectifying K+ channels and aquaporins 4 and 5 in autoimmune uveitis indicates misbalance in Müller glial cell-dependent ion and water homeostasis.

    Science.gov (United States)

    Eberhardt, Christina; Amann, Barbara; Feuchtinger, Annette; Hauck, Stefanie M; Deeg, Cornelia A

    2011-05-01

    Reactive gliosis is a well-established response to virtually every retinal disease. Autoimmune uveitis, a sight threatening disease, is characterized by recurrent relapses through autoaggressive T-cells. The purpose of this study was to assess retinal Müller glial cell function in equine recurrent uveitis (ERU), a spontaneous disease model resembling the human disease, by investigating membrane proteins implicated in ion and water homeostasis. We found that Kir2.1 was highly expressed in diseased retinas, whereas Kir4.1 was downregulated in comparison to controls. Distribution of Kir2.1 appeared Müller cell associated in controls, whereas staining of cell somata in the inner nuclear layer was observed in uveitis. In contrast to other subunits, Kir4.1 was evenly expressed along equine Müller cells, whereas in ERU, Kir4.1 almost disappeared from Müller cells. Hence, we suggest a different mechanism for potassium buffering in the avascular equine retina and, moreover, an impairment in uveitis. Uveitic retinas showed significantly increased expression of AQP4 as well as a displaced expression from Müller cells in healthy specimens to an intense circular expression pattern in the outer nuclear layer in ERU cases. Most interestingly, we detected the aquaporin family member protein AQP5 to be expressed in Müller cells with strong enrichments in Müller cell secondary processes. This finding indicates that fluid regulation within the equine retina may be achieved by an additional aquaporin. Furthermore, AQP5 was significantly decreased in uveitis. We conclude that the Müller cell response in autoimmune uveitis implies considerable changes in its potassium and water physiology.

  20. Geomagnetic Indices Bulletin (GIB)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The Geomagnetic Indices Bulletin is a one page sheet containing the magnetic indices Kp, Ap, Cp, An, As, Am and the provisional aa indices. The bulletin is published...

  1. Candidate gene association study in type 2 diabetes indicates a role for genes involved in beta-cell function as well as insulin action.

    Directory of Open Access Journals (Sweden)

    Inês Barroso

    2003-10-01

    Full Text Available Type 2 diabetes is an increasingly common, serious metabolic disorder with a substantial inherited component. It is characterised by defects in both insulin secretion and action. Progress in identification of specific genetic variants predisposing to the disease has been limited. To complement ongoing positional cloning efforts, we have undertaken a large-scale candidate gene association study. We examined 152 SNPs in 71 candidate genes for association with diabetes status and related phenotypes in 2,134 Caucasians in a case-control study and an independent quantitative trait (QT cohort in the United Kingdom. Polymorphisms in five of 15 genes (33% encoding molecules known to primarily influence pancreatic beta-cell function-ABCC8 (sulphonylurea receptor, KCNJ11 (KIR6.2, SLC2A2 (GLUT2, HNF4A (HNF4alpha, and INS (insulin-significantly altered disease risk, and in three genes, the risk allele, haplotype, or both had a biologically consistent effect on a relevant physiological trait in the QT study. We examined 35 genes predicted to have their major influence on insulin action, and three (9%-INSR, PIK3R1, and SOS1-showed significant associations with diabetes. These results confirm the genetic complexity of Type 2 diabetes and provide evidence that common variants in genes influencing pancreatic beta-cell function may make a significant contribution to the inherited component of this disease. This study additionally demonstrates that the systematic examination of panels of biological candidate genes in large, well-characterised populations can be an effective complement to positional cloning approaches. The absence of large single-gene effects and the detection of multiple small effects accentuate the need for the study of larger populations in order to reliably identify the size of effect we now expect for complex diseases.

  2. The use of natural abundance stable isotopic ratios to indicate the presence of oxygen-containing chemical linkages between cellulose and lignin in plant cell walls.

    Science.gov (United States)

    Zhou, Youping; Stuart-Williams, Hilary; Farquhar, Graham D; Hocart, Charles H

    2010-06-01

    Qualitative and quantitative understanding of the chemical linkages between the three major biochemical components (cellulose, hemicellulose and lignin) of plant cell walls is crucial to the understanding of cell wall structure. Although there is convincing evidence for chemical bonds between hemicellulose and lignin and the absence of chemical bonds between hemicellulose and cellulose, there is no conclusive evidence for the presence of covalent bonds between cellulose and lignin. This is caused by the lack of selectivity of current GC/MS-, NMR- and IR-based methods for lignin characterisation as none of these techniques directly targets the possible ester and ether linkages between lignin and cellulose. We modified the widely-accepted "standard" three-step extraction method for isolating cellulose from plants by changing the order of the steps for hemicellulose and lignin removal (solubilisation with concentrated NaOH and oxidation with acetic acid-containing NaClO(2), respectively) so that cellulose and lignin could be isolated with the possible chemical bonds between them intact. These linkages were then cleaved with NaClO(2) reagent in aqueous media of contrasting (18)O/(16)O ratios. We produced cellulose with higher purity (a lower level of residual hemicellulose and no detectable lignin) than that produced by the "standard" method. Oxidative artefacts may potentially be introduced at the lignin removal stage; but testing showed this to be minimal. Cellulose samples isolated from processing plant-derived cellulose-lignin mixtures in media of contrasting (18)O/(16)O ratios were compared to provide the first quantitative evidence for the presence of oxygen-containing ester and ether bonds between cellulose and lignin in Zea mays leaves. However, no conclusive evidence for the presence or lack of similar bonds in Araucaria cunninghamii wood was obtained. Copyright 2010 Elsevier Ltd. All rights reserved.

  3. Immunonucleochemistry: a new method for in situ detection of antigens in the nucleus of cells in culture.

    Science.gov (United States)

    Moon, Il Soo; Lee, Hyunsook; Park, Sung Dong; Seog, Dae-Hyun

    2010-04-01

    The advancement of immunocytochemistry (ICC) allows one to observe detailed spatial distribution of cellular antigens, but, with some limitations. Using conventional ICC, it is difficult to distinguish the nuclear localization from cytoplasm, as two large subcellular compartments overlap on the z-axis. In this study, we have investigated whether in situ immunostaining of 'naked' nuclei could provide an unambiguous method for detection of nuclear antigens. We have designed a protocol that efficiently lyses plasmalemma, while keeping the nuclear envelope intact. The optimal condition for lysing the plasmalemma was 0.5% Nonidet P-40 for 5 min in both neuronal and non-neuronal cultured cells. Using this protocol, we could unambiguously isolate nuclear from cytoplasmic ICC signals. Since the present protocol has been designed for immunostaining of 'naked' nuclei from cultured or isolated cells, we have coined a new term to refer to this procedure as 'immunonucleochemistry' ('INC' for abbreviation).

  4. Red Cell Morphology Interference in Platelet Indices from Patients with Anemia and Low Mean Corpuscular Volume from the IPS Universitaria, Clínica León XIII

    OpenAIRE

    Jaramillo Arbeláez, Patricia E.; Universidad de Antioquia-Clínica León XIII; Acevedo Toro, Paola A.; Universidad de Antioquia

    2013-01-01

    Introduction: The microcytosis and poikilocytosis interfere with platelet count in microcytic anemia, which alters the platelet indices determination in hematology analyzers with impedance.Objective: To evaluate the red cell morphology in blood smears to identify those alterations that interfere with platelet indices results in patients with microcytic anemia. Materials and Methods: Fifty adult patients hospitalized with heterogeneous microcytic anemia were confirmed through automated full bl...

  5. Impaired CD4-cell immune reconstitution upon HIV therapy in patients with toxoplasmic encephalitis compared to patients with pneumocystis pneumonia as AIDS indicating disease

    Directory of Open Access Journals (Sweden)

    Kastenbauer U

    2009-06-01

    Full Text Available Abstract Objectives There is only little data on immune reconstitution in antiretroviral naïve AIDS-patients with toxoplasmosis. The observation of several cases with reduced increase of CD4-cells upon start of antiretroviral treatment (ART prompted us to investigate the topic using the ClinSurv cohort. Methods 17 German HIV treatment centers contribute to ClinSurv, a multicentre observational cohort under the auspices of the Robert Koch Institute. We retrospectively selected all antiretroviral-naïve patients with toxoplasmic encephalitis (TE and -as comparator group -with pneumocystosis (PCP between January 1999 and December 2005. Results A total of 257 patients were included in the analysis, 61 with TE and 196 with PCP. Demographic baseline data showed differences with regard to gender, transmission group, and baseline CD4+ counts (60.9 vs. 44.7/μl, p = 0.022. After ART-initiation the increase in CD4+ lymphocytes was lower in the TE-versus the PCP-group in the first, second and fourth three-month-period (74.4 vs. 120.3/μl, p = 0.006; 96.6 vs. 136.2/μl, p = 0.021; 156.5 vs. 211.5/μl, p = 0.013. Viral load (VL was higher in the PCP-group at baseline (4.46 log10cop/ml vs. 5.00 log10cop/ml, p = 0.008, while virological success of ART was equal. Conclusions Our data show for the first time that the average CD4+ T-cell increase of patients with toxoplasmosis is impaired compared to PCP-patients. Most clinicians would not be prepared to discontinue follow-up TE-therapy unless CD4+ counts of 200/μl are reached. Explanation for our finding might be the myelosuppressive side effect of pyrimethamine, possible interactions of toxoplasmosis therapy with ART, or an unknown direct biological influence of toxoplasmosis on immune restoration.

  6. Effect of human umbilical cord mesenchymal stem cell adjuvant therapy on liver function and fibrosis indicators as well as the degree of inflammation in patients with hepatitis B cirrhosis

    Institute of Scientific and Technical Information of China (English)

    Gui-Jin Luo; Ping-Guang Lei

    2016-01-01

    Objective:To analyze the effect of human umbilical cord mesenchymal stem cell adjuvant therapy on liver function and fibrosis indicators as well as the degree of inflammation in patients with hepatitis B cirrhosis.Methods:A total of 80 cases with hepatitis B cirrhosis in our hospital from August 2012 to November 2014 were included for study. According to different treatment methods, all included patients were divided into observation group and control group by half. Control group received conventional treatment, observation group received human umbilical cord mesenchymal stem cell adjuvant therapy, and then differences in the levels of liver function indicators, liver fibrosis indicators, inflammation-related indicators and illness-related indicators were compared between two groups.Results:Serum ALB, GLB and A/G values of observation group after treatment were higher than those of control group, andα2-M, TB, APO-A1 and GGT values were lower than those of control group (P<0.05); serum HA, LN, CIV, PⅢNP and PLD values of observation group after treatment were lower than those of control group (P<0.05); serum TGF-β1, PCT, WBC and SIL-2R levels of observation group after treatment were lower than those of control group (P<0.05); serum FT3 and ADP values of observation group after treatment were higher than those of control group, and NO, EGF, ADM and IR values were lower than those of control group (P<0.05).Conclusions:Human umbilical cord mesenchymal stem cell adjuvant therapy for patients with hepatitis B cirrhosis can optimize liver function and inhibit disease progression, and it has active clinical significance.

  7. Poor visualization of renal collecting system in intravenous urography as an indicator of invasive transitional cell carcinoma in the upper urinary tract

    Institute of Scientific and Technical Information of China (English)

    SHEN Zhou-jun; LI Liao-yuan; LIAO Guo-dong; CHEN Dong

    2007-01-01

    Background Transitional cell carcinoma of the upper urinary tract (UUT-TCC) accounts for 5% to 10% of all renal tumours and 5% to 6% of all urothelial tumours all over the world. In China, the proportion of UUT-TCC to all urothelial tumours may be 26%, which is higher than that in the western world. The early diagnosis of UUT-TCC is difficult and the present study elucidates the diagnostic value of poor or nonvisualization (PNV) in intravenous urography in patients with UUT-TCC and its correlations with pathological findings and clinical characteristics.Methods The data of 172 consecutive patients between January 1997 and January 2005 with UUT-TCC who underwent nephroureterectomy in our departments were selected and analyzed retrospectively.Results Of our sample, 144 cases presented with gross haematuria (83.7%) and 12 with microscopic haematuria (7.0%). Forty-six cases (26.7%) were detectable by cytology. Filling defect identified 36 positive cases of 172 patients (20.9%), PNV was present in the images of 105 of 172 patients (61.0%). The detection rate by PNV (61.0%) was significantly different from that by cytology (26.7%) or by filling defect (20.9%) (P=0.031, P=0.001, respectively).Univariate logistic regression analysis for PNV showed that tumour stage, grade and size were significant predictors (P=0.028; P=0.031; P=0.006, respectively). Tumour stage and size were identified as independent risk factors in the multivariate logistic regression model (P=0.042; P=0.014).Conclusions Except for suspected urolithiasis, urinary tuberculosis or congenital abnormalities, UUT-TCC should be considered if PNV exists in intravenous urography especially of old patients. The value of PNV is much more significant than filling defect in intravenous urography in the diagnosis of UUT-TCC. It is supposed that PNV carries more risk of higher stage and larger tumour size in UTT-TCC.

  8. Observations on continuously growing roots of the sloth and the K14-Eda transgenic mice indicate that epithelial stem cells can give rise to both the ameloblast and root epithelium cell lineage creating distinct tooth patterns.

    Science.gov (United States)

    Tummers, Mark; Thesleff, Irma

    2008-01-01

    Root development is traditionally associated with the formation of Hertwig's epithelial root sheath (HERS), whose fragments give rise to the epithelial cell rests of Malassez (ERM). The HERS is formed by depletion of the core of stellate reticulum cells, the putative stem cells, in the cervical loop, leaving only a double layer of the basal epithelium with limited growth capacity. The continuously growing incisor of the rodent is subdivided into a crown analog half on the labial side, with a cervical loop containing a large core of stellate reticulum, and its progeny gives rise to enamel producing. The lingual side is known as the root analog and gives rise to ERM. We show that the lingual cervical loop contains a small core of stellate reticulum cells and suggest that it acts as a functional stem cell niche. Similarly we show that continuously growing roots represented by the sloth molar and K14-Eda transgenic incisor maintain a cervical loop with a small core of stellate reticulum cells around the entire circumference of the tooth and do not form a HERS, and still give rise to ERM. We propose that HERS is not a necessary structure to initiate root formation. Moreover, we conclude that crown vs. root formation, i.e. the production of enamel vs. cementum, and the differentiation of the epithelial cells into ameloblasts vs. ERM, can be regulated independently from the regulation of stem cell maintenance. This developmental flexibility may underlie the developmental and evolutionary diversity in tooth patterning.

  9. Cytotoxic T cell response against the chimeric ETV6-AML1 protein in childhood acute lymphoblastic leukemia.

    Science.gov (United States)

    Yotnda, P; Garcia, F; Peuchmaur, M; Grandchamp, B; Duval, M; Lemonnier, F; Vilmer, E; Langlade-Demoyen, P

    1998-07-15

    Cytotoxic T lymphocytes (CTL) are potent effector cells that could provide long term antitumor immunity if induced by appropriate vaccines. CTL recognize 8-14 amino acid-long peptides processed intracellularly and presented by MHC class I molecules. A well-characterized example of a potential tumor antigen in childhood pre-B Acute Lymphoblastic Leukemia (ALL) results from the chromosomal translocation 12;21 leading to the fusion of the ETV6 and AML1 genes. This translocation is observed in > 25% of ALL-patients. In this study, we have examined whether the chimeric ETV6-AML1 protein could serve as a tumor specific antigen for CTL in HLA-A2.1 individuals. We have identified a nonapeptide (RIAECILGM), encoded by the fusion region of the ETV6-AML1 protein, that binds to HLA-A2.1 molecules and induces specific primary CTL in peripheral blood lymphocytes from healthy donors. These CTL specifically lysed HLA-A2.1 tumor cells endogeneously expressing the ETV6-AML fusion protein. CTL with similar functional capacities were found with high frequencies and cloned from one patient's bone marrow indicating that ETV6-AML1-specific anti-ALL CTL are, at least in some patients, spontaneously stimulated and might participate to host antileukemia defense.

  10. Microphotographs of cyanobacteria documenting the effects of various cell-lysis techniques

    Science.gov (United States)

    Rosen, Barry H.; Loftin, Keith A.; Smith, Christopher E.; Lane, Rachael F.; Keydel, Susan P.

    2011-01-01

    Cyanotoxins are a group of organic compounds biosynthesized intracellularly by many species of cyanobacteria found in surface water. The United States Environmental Protection Agency has listed cyanotoxins on the Safe Drinking Water Act's Contaminant Candidate List 3 for consideration for future regulation to protect public health. Cyanotoxins also pose a risk to humans and other organisms in a variety of other exposure scenarios. Accurate and precise analytical measurements of cyanotoxins are critical to the evaluation of concentrations in surface water to address the human health and ecosystem effects. A common approach to total cyanotoxin measurement involves cell membrane disruption to release the cyanotoxins to the dissolved phase followed by filtration to remove cellular debris. Several methods have been used historically, however no standard protocols exist to ensure this process is consistent between laboratories before the dissolved phase is measured by an analytical technique for cyanotoxin identification and quantitation. No systematic evaluation has been conducted comparing the multiple laboratory sample processing techniques for physical disruption of cell membrane or cyanotoxins recovery. Surface water samples collected from lakes, reservoirs, and rivers containing mixed assemblages of organisms dominated by cyanobacteria, as well as laboratory cultures of species-specific cyanobacteria, were used as part of this study evaluating multiple laboratory cell-lysis techniques in partnership with the U.S. Environmental Protection Agency. Evaluated extraction techniques included boiling, autoclaving, sonication, chemical treatment, and freeze-thaw. Both treated and untreated samples were evaluated for cell membrane integrity microscopically via light, epifluorescence, and epifluorescence in the presence of a DNA stain. The DNA stain, which does not permeate live cells with intact membrane structures, was used as an indicator for cyanotoxin release into the

  11. Gamma-Glutamylcysteine Inhibits Oxidative Stress in Human Endothelial Cells

    Science.gov (United States)

    2012-01-01

    GGC treatment and pooled. Then, cells were suspended and lysed with hypotonic buffer and 10% (w/v) Nonidet P - 40 . After spinning, the cell pellet was...correlation with PPARγ DNA binding levels (pb0.05) and positive correlation trends with NF-κB p65 DNA binding ( p =0.059), 8-epi- PGF2α, ( p =0.080), and...TBARS ( p =0.129) levels through Pearson’s correlations. GSH synthetase (GSS) expression We found statistically lower GSS protein levels (0.93 to 0.85

  12. Lactobacilli Modulate Natural Killer Cell Responses In Vitro

    DEFF Research Database (Denmark)

    Fink, Lisbeth Nielsen; Christensen, Hanne Risager; Frøkiær, Hanne

    Natural killer (NK) cells are cells of the non-specific immune system lysing altered self-cells. A non-cytolytic subset of NK cells may serve a regulatory role by secreting cytokines. Bacteria translocating across the gastrointestinal mucosa are presumed to gain access to NK cells, as consumption...... of certain lactic acid bacteria has been shown to increase in vivo NK cytotoxicity. Here, we investigated how human gut flora-derived lactobacilli affect NK cells in vitro, by measuring proliferation and IFN-gamma production of human NK cells upon bacterial stimulation. CD3-CD56+ NK cells were isolated from...... buffy coats by negative isolation using non-NK lineage specific antibodies and magnetic beads. NK cells were incubated with 10 microg/ml UV-inactivated bacteria or 10 microg/ml phytohemagglutinin (PHA) for four days. Proliferation was assessed by incorporation of radioactive thymidine into NK cell DNA...

  13. In vitro enhancement of extracellular matrix formation as natural bioscaffold for stem cell culture

    Science.gov (United States)

    Naroeni, Aroem; Shalihah, Qonitha; Meilany, Sofy

    2017-02-01

    Growing cells in plastic with liquid media for in vitro study is very common but far from physiological. The use of scaffold materials is more biocompatible. Extracellular matrix provides tissue integrity which acts as a native scaffold for cell attachment and interaction, as well as it serves as a reservoir for growth factors. For this reason, we have developed natural scaffold from mice fibroblast to form a natural scaffold for stem cell culture. Fibroblasts were cultured under crowded condition and lysed to form natural scaffold. The natural scaffold formation was observed using immunofluorescence which then will be used and tested for stem cell propagation and differentiation.

  14. CREDIT Performance Indicator Framework

    DEFF Research Database (Denmark)

    Frandsen, Anne Kathrine; Bertelsen, Niels Haldor; Haugbølle, Kim

    2010-01-01

    During the past two years the Nordic Baltic research project CREDIT (Construction and Real Estate – Developing Indicators for Transparency) has worked with the aim to improve transparency of value creation in building and real estate. One of the central deliverables of the CREDIT project...... was a framework of indicators relevant in building and real estate and applicable in the Nordic and Baltic countries as well as a proposal for a set of key indicators. The study resulting in CREDIT Performance Indicator Framework has been based on 28 case studies of evaluation practises in the building and real...... regulations in the countries participating in CREDIT. The Performance Indicator Framework encompassed 187 indicators grouped in 7 main groups of indicators and 42 sub-groups. Based on the CREDIT case studies it was concluded that there neither is link between certain indicators and specific building types...

  15. Geomagnetic aa Indices

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The geomagnetic aa indices are the continuation of the series beginning in the year 1868. A full description of these indices is given in the International...

  16. Altimeter Setting Indicator

    Data.gov (United States)

    Department of Transportation — The Altimeter Setting Indicator (ASI) is an aneroid system used at airports to provide an altimeter setting for aircraft altimeters. This indicator may be an analog...

  17. Alternative Solar Indices

    Energy Technology Data Exchange (ETDEWEB)

    Lantz, L.J.

    1980-07-01

    Possible alternative Solar Indices which could either be a perturbation from the currently defined Solar Index or possible indices based on current technologies for other media markets are discussed. An overview is given of the current project, including the logic that was utilized in defining its current structure and then alternative indices and definitions are presented and finally, recommendations are made for adopting alternative indices.

  18. A t (11; 22 (p13; q12 EWS-WT 1 positive desmoplastic small round cell tumor of the maxilla: An unusual case indicating the role of molecular diagnosis in round cell sarcomas

    Directory of Open Access Journals (Sweden)

    Rekhi B

    2010-01-01

    Full Text Available A desmoplastic small round cell tumor (DSRCT is an uncommon tumor characterized by polyphenotypic expression and a specific reciprocal translocation t (11; 22 (p13; q12. It has been rarely identified in the head and neck region. Herein, we describe a DSRCT in the maxilla of a young man, who was initially diagnosed with a primitive neuroectodermal tumor (PNET, based on histopathological appearance of a round cell tumor, with MIC2 and -FLI-1 positivity, on immunohistochemistry (IHC. Diagnosis of a DSRCT was confirmed on molecular analysis with positive -RT-PCR and sequencing results for EWS-WT1 transcript and negativity for EWS-FL1. The case is presented to highlight the value of molecular diagnosis in round cell sarcomas at uncommon sites, especially when similar IHC markers can be expressed in a PNET and a DSRCT. An exact diagnosis of a round cell sarcoma has a therapeutic relevance.

  19. Epitope distance to the target cell membrane and antigen size determine the potency of T cell-mediated lysis by BiTE antibodies specific for a large melanoma surface antigen.

    Science.gov (United States)

    Bluemel, Claudia; Hausmann, Susanne; Fluhr, Petra; Sriskandarajah, Mirnalini; Stallcup, William B; Baeuerle, Patrick A; Kufer, Peter

    2010-08-01

    Melanoma chondroitin sulfate proteoglycan (MCSP; also called CSPG4, NG2, HMW-MAA, MSK16, MCSPG, MEL-CSPG, or gp240) is a surface antigen frequently expressed on human melanoma cells, which is involved in cell adhesion, invasion and spreading, angiogenesis, complement inhibition, and signaling. MCSP has therefore been frequently selected as target antigen for development of antibody- and vaccine-based therapeutic approaches. We have here used a large panel of monoclonal antibodies against human MCSP for generation of single-chain MCSP/CD3-bispecific antibodies of the BiTE (for bispecific T cell engager) class. Despite similar binding affinity to MCSP, respective BiTE antibodies greatly differed in their potency of redirected lysis of CHO cells stably transfected with full-length human MCSP, or with various MCSP deletion mutants and fusion proteins. BiTE antibodies binding to the membrane proximal domain D3 of MCSP were more potent than those binding to more distal domains. This epitope distance effect was corroborated with EpCAM/CD3-bispecific BiTE antibody MT110 by testing various fusion proteins between MCSP and EpCAM as surface antigens. CHO cells expressing small surface target antigens were generally better lysed than those expressing larger target antigens, indicating that antigen size was also an important determinant for the potency of BiTE antibody. The present study for the first time relates the positioning of binding domains and size of surface antigens to the potency of target cell lysis by BiTE-redirected cytotoxic T cells. In case of the MCSP antigen, this provides the basis for selection of a maximally potent BiTE antibody candidate for development of a novel melanoma therapy.

  20. Activation of p44/42 MAPK plays a role in the TBT-induced loss of human natural killer (NK) cell function.

    Science.gov (United States)

    Dudimah, Fred D; Griffey, Denisha; Wang, Xiaofei; Whalen, Margaret M

    2010-10-01

    Natural killer (NK) cells destroy (lyse) tumor cells, virally infected cells, and antibody-coated cells. Previous studies indicated that exposure to the environmental contaminant tributyltin (TBT) decreases the lytic function of NK cells and activates mitogen-activated protein kinases (MAPK), including p44/42 (Aluoch and Whalen Toxicology 209:263-277, 2005). If activation of p44/42 is required for TBT-induced decreases of lytic function, then activation of p44/42 to similar extents by pharmacological agents such as phorbol 12-myristate 13-acetate (PMA) should mimic to some extent changes induced in NK cells with TBT exposures. NK cells were exposed to PMA concentrations between 0.25 and 10 nM for 10 min, 1 h, and 6 h before determining the lytic function ((51)Cr release assay) and phosphorylation state of MAPKs (Western blot). A 1-h exposure of NK cells to 5 nM PMA resulted in a loss of lytic function of 47%. Western blot analysis showed that a 1-h exposure to 5 nM PMA caused a sixfold increase in phospho-p44/42 levels. Previous studies showed a fivefold increase in phospho-p44/42 in response to a 1-h exposure to 300 nM TBT. Exposure to 300 nM TBT caused about a 40% decrease in lytic function. This study supports the hypothesis that p44/42 activation (as seen with TBT exposures) can cause a loss of NK-cell lytic function.

  1. Cells

    Directory of Open Access Journals (Sweden)

    Zhao-Hui Jin

    2012-11-01

    Full Text Available As cancer stem cells (CSCs are postulated to play critical roles in cancer development, including metastasis and recurrence, CSC imaging would provide valuable information for cancer treatment and lead to CSC-targeted therapy. To assess the possibility of in vivo CSC targeting, we conducted basic studies on radioimmunotargeting of cancer cells positive for CD133, a CSC marker recognized in various cancers. Antibodies against CD133 were labeled with 125I, and their in vitro cell binding properties were tested. Using the same isotype IgG as a control, in vivo biodistribution of the labeled antibody retaining immunoreactivity was examined in mice bearing an HCT116 xenograft in which a population of the cancer cells expressed CD133. Intratumoral distribution of the labeled antibody was examined and compared to the CD133 expression pattern. The 125I-labeled anti-CD133 antibody showed a modest but significantly higher accumulation in the HCT116 xenograft compared to the control IgG. The intratumoral distribution of the labeled antibody mostly overlapped with the CD133 expression, whereas the control IgG was found in the area close to the necrotic tumor center. Our results indicate that noninvasive in vivo targeting of CSCs could be possible with radiolabeled antibodies against cell membrane markers.

  2. eHealth indicators

    DEFF Research Database (Denmark)

    HYPPÖNEN, Hannele; AMMENWERTH, Elske; Nøhr, Christian;

    2012-01-01

    eHealth indicators are needed to measure defined aspects of national eHealth implementations. However, until now, eHealth indicators are ambiguous or unclear. Therefore, an expert workshop "Towards an International Minimum Dataset for Monitoring National Health Information System Implementations......" was organized. The objective was to develop ideas for a minimum eHealth indicator set. The proposed ideas for indicators were classified based on EUnetHTA and De-Lone & McClean, and classification was compared with health IT evaluation criteria classification by Ammenwerth & Keizer. Analysis of the workshop...... results emphasized the need for a common methodological framework for defining and classifying eHealth indicators. It also showed the importance of setting the indicators into context. The results will benefit policy makers, developers and researchers in pursuit of provision and use of evidence...

  3. Indicators for environmental sustainability

    DEFF Research Database (Denmark)

    Dong, Yan; Hauschild, Michael Zwicky

    2017-01-01

    Decision making on sustainable consumption and production requires scientifically based information on sustainability. Different environmental sustainability targets exist for specific decision problems. To observe how well these targets are met, relevant environmental indicators are needed....... In this study, we reviewed indicators applied in life cycle assessment (LCA), planetary boundary framework (PB), and Sustainable Development Goals (SDGs) developed under United Nation. The aim is to 1) identify their applications and relevant decision context; 2) Review their indicators and categorize them...

  4. Indicators for environmental sustainability

    DEFF Research Database (Denmark)

    Dong, Yan; Hauschild, Michael Zwicky

    2017-01-01

    Decision making on sustainable consumption and production requires scientifically based information on sustainability. Different environmental sustainability targets exist for specific decision problems. To observe how well these targets are met, relevant environmental indicators are needed....... In this study, we reviewed indicators applied in life cycle assessment (LCA), planetary boundary framework (PB), and Sustainable Development Goals (SDGs) developed under United Nation. The aim is to 1) identify their applications and relevant decision context; 2) Review their indicators and categorize them...

  5. A cell extraction method for oily sediments

    Directory of Open Access Journals (Sweden)

    Michael eLappé

    2011-11-01

    Full Text Available Hydrocarbons can be found in many different habitats and represent an important carbon source for microbes. As fossil fuels, they are also an important economical resource, through natural seepage or accidental release they can also be major pollutants. DNA-specific stains and molecular probes bind to hydrocarbons, causing massive background fluorescence and thereby hampering cell enumeration. The cell extraction procedure of Kallmeyer et al. (2008 separates the cells from the sediment matrix. In principle, this technique can also be used to separate cells from oily sediments, but it is not optimized for this application.Here we present a modified extraction method in which the hydrocarbons are removed prior to cell extraction. Due to the reduced background fluorescence the microscopic image becomes clearer, making cell identification and enumeration much easier. Consequently, the resulting cell counts from samples treated according to our new protocol are significantly higher than those treated according to Kallmeyer et al. (2008. We tested different amounts of a variety of solvents for their ability to remove hydrocarbons and found that n-hexane and – in samples containing more biodegraded oils – methanol, delivered the best results. However, as solvents also tend to lyse cells, it was important to find the optimum solvent to sample ratio, at which hydrocarbon extraction is maximised and cell lysis minimized. A ratio between slurry and solvent of 1:2 to 1:5 delivered the highest cell counts without lysing too many cells. The method provided reproducibly good results on samples from very different environments, both marine and terrestrial.

  6. Data Quality and Indicators

    Directory of Open Access Journals (Sweden)

    Jorge Matute

    2007-01-01

    Full Text Available This study highlights the importance of collecting good quality data from multidisciplinary studies. Bias in data may be the result of instrument inaccuracies, imprecise data recording techniques, inaccurate data entry to computers or inappropriate statistical analysis and presentation. Recommendations for good data quality control are given. Different types of data are discussed: raw data, simple indicators and complex indicators. It is shown how measurements from the components of multidisciplinary systems can be combined to form complex indicators and a specific example is given using Z-scores and dot charts. Finally the accumulated effect of bias in the individual component measurements upon the combined indicator is shown.

  7. Scopus' SNIP Indicator

    CERN Document Server

    Leydesdorff, Loet

    2010-01-01

    Rejoinder to Moed [arXiv:1005.4906]: Our main objection is against developing new indicators which, like some of the older ones (for example, the "crown indicator" of CWTS), do not allow for indicating error because they do not provide a statistics, but are based, in our opinion, on a violation of the order of operations. The claim of validity for the SNIP indicator is hollow because the normalizations are based on field classifications which are not valid. Both problems can perhaps be solved by using fractional counting.

  8. CREDIT Performance Indicator Framework

    DEFF Research Database (Denmark)

    Frandsen, Anne Kathrine; Bertelsen, Niels Haldor; Haugbølle, Kim

    2010-01-01

    During the past two years the Nordic Baltic research project CREDIT (Construction and Real Estate – Developing Indicators for Transparency) has worked with the aim to improve transparency of value creation in building and real estate. One of the central deliverables of the CREDIT project was a fr......During the past two years the Nordic Baltic research project CREDIT (Construction and Real Estate – Developing Indicators for Transparency) has worked with the aim to improve transparency of value creation in building and real estate. One of the central deliverables of the CREDIT project...... was a framework of indicators relevant in building and real estate and applicable in the Nordic and Baltic countries as well as a proposal for a set of key indicators. The study resulting in CREDIT Performance Indicator Framework has been based on 28 case studies of evaluation practises in the building and real...... regulations in the countries participating in CREDIT. The Performance Indicator Framework encompassed 187 indicators grouped in 7 main groups of indicators and 42 sub-groups. Based on the CREDIT case studies it was concluded that there neither is link between certain indicators and specific building types...

  9. Climate Change Indicators

    Science.gov (United States)

    Presents information, charts and graphs showing measured climate changes across 40 indicators related to greenhouse gases, weather and climate, oceans, snow and ice, heath and society, and ecosystems.

  10. Bayesian Sampling using Condition Indicators

    DEFF Research Database (Denmark)

    Faber, Michael H.; Sørensen, John Dalsgaard

    2002-01-01

    . This allows for a Bayesian formulation of the indicators whereby the experience and expertise of the inspection personnel may be fully utilized and consistently updated as frequentistic information is collected. The approach is illustrated on an example considering a concrete structure subject to corrosion....... It is shown how half-cell potential measurements may be utilized to update the probability of excessive repair after 50 years....

  11. Integrative analyses of gene expression and DNA methylation profiles in breast cancer cell line models of tamoxifen-resistance indicate a potential role of cells with stem-like properties

    DEFF Research Database (Denmark)

    Lin, Xue; Li, Jian; Yin, Guangliang

    2013-01-01

    Development of resistance to tamoxifen is an important clinical issue in the treatment of breast cancer. Tamoxifen resistance may be the result of acquisition of epigenetic regulation within breast cancer cells, such as DNA methylation, resulting in changed mRNA expression of genes pivotal...

  12. Indicators of sustainable tourism

    Directory of Open Access Journals (Sweden)

    Jovičić Dobrica

    2010-01-01

    Full Text Available The indicators, precisely describing the linkages between tourism and the environment, social and cultural base, are not easily available. How ever, some relevant organizations (WTO, EU, OECD, etc., institutions and experts, have been hardly working to create the indicators of sustainable tourism. Whereas the economic objectives are easily defined by the use of the traditional indicators used in national and business economics, it is very difficult to identify widely applicable environmental, social and cultural indicators. In order to stimulate and alleviate the process of sustainable tourism development, EU created the list of comparative indicators of sustainable tourism. In preparing this list, special attention is paid to identification of valid indicators of real tourism impacts on the social and cultural environment (the entire set of traditions, customs, history, hospitality and culture that characterize a given area, that is a very complex task. Assuming the fact that the related indicators have been analyzed in many European countries, this paper is focused on applying the related indicators in research of tourism development in villages of the Kosjerić community. .

  13. Indicators of coal metamorphism

    Energy Technology Data Exchange (ETDEWEB)

    Proskuryakov, A.E.

    1982-06-01

    Important in determining metamorphism of coal is the reliability of indicators of coalification. Both the reflection of vitrinite and emission of volatile matter have been used for this purpose. To determine which indicator more accurately characterizes metamorphism of coal, their conformity to the following demands was established: 1. uniformity in direction of change of parameters with degree of metamorphism; 2. independence of the indicator of the genetic characteristics of coal (petrographic composition, reduction and oxidation of coal); 3. sensitivity of indicator. Both indicators conform to the first requirement. Emission of volatile substance decreases and reflective capacity of vitrinite increases uniformly with degree of metamorphism. However, the reflectivity of vitrinite is not influenced by petrographic composition of coals and is less dependent on the oxidation and reduction of coal than emission of volatile matter. It is also a more sensitive indicator distinguishing more degrees of metamorphism than emission of volatile matter. Reflectivity of vitrinite is a more reliable indicator of metamorphism than emission of volatile matter. However, in many laboratories this indicator is not measured with sufficient accuracy. To correct this, measuring equipment must be standardized.

  14. Key performance indicators

    NARCIS (Netherlands)

    Zwetsloot, G.I.J.M.

    2014-01-01

    This paper addresses how organisations can use OSH performance indicators. This is an important way to mainstream OSH into business management. Key performance indicators (KPIs) should provide objective data on the OSH situation. It is often said that ‘what gets measured gets managed’. Without

  15. Key performance indicators

    NARCIS (Netherlands)

    Zwetsloot, G.I.J.M.

    2014-01-01

    This paper addresses how organisations can use OSH performance indicators. This is an important way to mainstream OSH into business management. Key performance indicators (KPIs) should provide objective data on the OSH situation. It is often said that ‘what gets measured gets managed’. Without infor

  16. Targeting Cancer Stem Cells with Natural Killer Cell Immunotherapy.

    Science.gov (United States)

    Luna, Jesus I; Grossenbacher, Steven K; Murphy, William J; Canter, Robert J

    2017-03-01

    Standard cytoreductive cancer therapy, such as chemotherapy and radiotherapy, are frequently resisted by a small portion of cancer cells with 'stem-cell' like properties including quiescence and repopulation. Immunotherapy represents a breakthrough modality for improving oncologic outcomes in cancer patients. Since the success of immunotherapy is not contingent on target cell proliferation, it may also be uniquely suited to address the problem of resistance and repopulation exerted by cancer stem cells (CSCs). Areas covered: Natural killer (NK) cells have long been known for their ability to reject allogeneic hematopoietic stem cells, and there are increasing data demonstrating that NK cells can selectively identify and lyse CSCs. The authors review the current knowledge of CSCs and NK cells and highlight recent studies that support the concept that NK cells are capable of targeting CSC in solid tumors, especially in the context of combination therapy simultaneously targeting non-CSCs and CSCs. Expert opinion: Unlike cytotoxic cancer treatments, NK cells can target and eliminate quiescent/non-proliferating cells such as CSCs, and these enigmatic cells are an important source of relapse and metastasis. NK targeting of CSCs represents a novel and potentially high impact method to capitalize on the intrinsic therapeutic potential of NK cells.

  17. VP Ellipsis without Indices

    DEFF Research Database (Denmark)

    Hardt, Daniel; Asher, Nicholas; Hunter, Julie

    2013-01-01

    This paper compares two views on the status of indices in syntactic and logical representations. On a structural view, indices are syntactic formants on a par with node labels and phrase bracketings, and are thus a part of the logical forms that are derived from syntactic representations....... On the process view, an index is not a syntactic object at all, but rather, an indication of the output of a resolution process.In this paper we argue that a recent body of data provides a clear empirical basis for distinguishing between these two views of indices. We argue that cases of sloppy VP ellipsis pose...... insurmountable problems for the structural view of indices, while these problems do not arise for the process view. Furthermore, we show that this resolution process is constrained by the semantics of various discourse relations....

  18. Evaluating Living Standard Indicators

    Directory of Open Access Journals (Sweden)

    Birčiaková Naďa

    2015-09-01

    Full Text Available This paper deals with the evaluation of selected available indicators of living standards, divided into three groups, namely economic, environmental, and social. We have selected six countries of the European Union for analysis: Bulgaria, the Czech Republic, Hungary, Luxembourg, France, and Great Britain. The aim of this paper is to evaluate indicators measuring living standards and suggest the most important factors which should be included in the final measurement. We have tried to determine what factors influence each indicator and what factors affect living standards. We have chosen regression analysis as our main method. From the study of factors, we can deduce their impact on living standards, and thus the value of indicators of living standards. Indicators with a high degree of reliability include the following factors: size and density of population, health care and spending on education. Emissions of carbon dioxide in the atmosphere also have a certain lower degree of reliability.

  19. Regulation of sterile α- and armadillo motif (SARM) containing protein expression in Pam2CSK4- and Pam3CSK4-activated mouse macrophage cell line (RAW264.7) requires TLR9.

    Science.gov (United States)

    Pudla, Matsayapan; Kulsantiwong, Panthong; Srisaowakarn, Chanya; Utaisincharoen, Pongsak

    2017-09-09

    We aimed to investigate the involvement of surface TLRs and endosomal TLRs in the regulation of SARM expression by TLR2 ligands (Pam2CSK4 and Pam3CSK4). Mouse macrophage cell line (RAW264.7) was treated with either Pam2CSK4 or Pam3CSK4 (TLR2 ligands) at a concentration of 100 ng/ml. At indicated time points, the treated cells were lysed. The gene and protein expression of SARM were determined by RT-PCR and immunoblotting, respectively. For silencing of TLR9 function, the cells were transfected with TLR9 siRNAs before stimulation by these two TLR2 ligands RESULTS: The SARM expression was upregulated at both transcriptional and translational levels in time-dependent manner during activation of Pam2CSK4 and Pam3CSK4 in mouse macrophages. Blocking of ligand internalization by cytochalasin D showed interference effect with SARM expression. Moreover, our results also demonstrated that endosomal acidification and TLR9 were required for SARM expression suggesting the essential role of endosomal compartment acidification and TLR9 in regulating SARM expression. Our findings suggested the collaboration of TLR2-TLR9 at least in the regulation of SARM expression. However, the underlying mechanism that participated in these two TLRs cooperation is underinvestigated.

  20. The radon indicator

    Science.gov (United States)

    Samuelsson, L.

    2005-11-01

    The radon indicator is an efficient instrument for measuring the radon daughter concentrations in a house or dwelling. Physics or environmental science students could build a radon indicator as a student project. Another possibility would be to use a radon indicator in a student investigation of radon levels in different houses. Finally the radon indicator is an excellent device for producing a radioactive source, free of charge, for the study of α-, β- and γ-radiation. The half-life of the activity collected is approximately 40 min. The radon indicator makes use of an electrostatic method by which charged particles are drawn to a small aluminium plate with a high negative voltage (-5 kV), thus creating a strong electric field between the plate and a surrounding copper wire. The radioactivity on the plate is subsequently measured by a GM-counter and the result calculated in Bq m-3. The collecting time is just 5.5 min and therefore the instrument is only suitable for use in a short-time method for indicating the radon concentration. An improved diagram, ground-radon and/or wall-radon in houses, is presented on the basis of the author's measurements recorded with the radon indicator over many years. This diagram is very useful when discussing how to reduce radiation levels in homes.

  1. Solar Indices Bulletin

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Solar Indices Bulletin is a prompt monthly information product that is distributed within two weeks after the observation month closes. For the month just ended,...

  2. Solar Indices - Solar Corona

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Collection includes a variety of indices related to solar activity contributed by a number of national and private solar observatories located worldwide. This...

  3. Indicators and SEA

    DEFF Research Database (Denmark)

    Gao, Jingjing; Kørnøv, Lone; Christensen, Per

    Abstract: Indicators are widely used in SEA to measure, communicate and monitor impacts from a proposed policy, plan or programme, and can improve the effectiveness for the SEA by simplifying the complexity of both assessment and presentation. Indicators can be seen as part of the implementation...... process helping to understand, communicate and, integrate important environmental issues in planning and decision-making. On the other hand, use of indicators can also limit SEA effectiveness, if the ones chosen are biased or limited, if the aggregation gives incorrect interpretation...... and if the information requirement for different target groups is not addressed. Indicators are widely used in SEA to measure, communicate and monitor impacts from a proposed policy, plan or programme, and can improve the effectiveness for the SEA by simplifying the complexity of both assessment and presentation...

  4. NOHSS Adult Indicators

    Data.gov (United States)

    U.S. Department of Health & Human Services — 2012-2014 (even years). Data from BRFSS for indicators of adult oral health for even years from 2012 through 2014. National estimates are represented by the median...

  5. NOHSS Child Indicators

    Data.gov (United States)

    U.S. Department of Health & Human Services — Data for School year-end 1994 through year-end 2017. State oral health surveys are the data sources for these indicators. States periodically conduct independent...

  6. NOHSS Child Indicators

    Data.gov (United States)

    U.S. Department of Health & Human Services — Data for School year-end 1994 through year-end 2016. State oral health surveys are the data sources for these indicators. States periodically conduct independent...

  7. Indicators: Sediment Enzymes

    Science.gov (United States)

    Sediment enzymes are proteins that are produced by microorganisms living in the sediment or soil. They are indicators of key ecosystem processes and can help determine which nutrients are affecting the biological community of a waterbody.

  8. NOHSS Child Indicators

    Data.gov (United States)

    U.S. Department of Health & Human Services — Data for School year-end 1994 through year-end 2016. State oral health surveys are the data sources for these indicators. States periodically conduct independent...

  9. NOHSS Adult Indicators

    Data.gov (United States)

    U.S. Department of Health & Human Services — 2012-2014 (even years). Data from BRFSS for indicators of adult oral health for even years from 2012 through 2014. National estimates are represented by the median...

  10. Fishery Performance Indicators

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Performance indicators for landings, effort, revenue and distribution of revenue are collected for various fisheries nation-wide. The fisheries include catch and...

  11. Solar Indices - Solar Flares

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Collection includes a variety of indices related to solar activity contributed by a number of national and private solar observatories located worldwide. This...

  12. Solar Indices - Sunspot Numbers

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Collection includes a variety of indices related to solar activity contributed by a number of national and private solar observatories located worldwide. This...

  13. Solar Indices - Plage Regions

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Collection includes a variety of indices related to solar activity contributed by a number of national and private solar observatories located worldwide. This...

  14. Solar Indices - Solar Irradiance

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Collection includes a variety of indices related to solar activity contributed by a number of national and private solar observatories located worldwide. This...

  15. Solar Indices - Solar Ultraviolet

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Collection includes a variety of indices related to solar activity contributed by a number of national and private solar observatories located worldwide. This...

  16. ENSO Indices and Analyses

    Institute of Scientific and Technical Information of China (English)

    WANG Zhiren; WU Dexing; CHEN Xue'en; QIAO Ran

    2013-01-01

    New ENSO indices were developed and the spatial variability and temporal evolution of ENSO were analyzed based on the new indices and modeling experiments,as well as multiple data resources.The new indices,after being defined,were validated with their good diagnostic characteristics and correlation with wind and SST.In the analysis after the definition and validation of the new indices,ENSO feedbacks from wind,heat fluxes,and precipitation were spatially and temporally examined in order to understand ENSO variability and evolution with some emphasized points such as the interaction among the feedbacks,the role of westerly wind bursts and the transformation between zonal and meridional circulations in an ENSO cycle,and the typical pattern of modern ENSO.

  17. Laterality and reproductive indices.

    Science.gov (United States)

    Kalichman, Leonid; Kobyliansky, Eugene

    2008-01-01

    Several previous studies support the association between manual dominance and age at menarche or age at menopause. The aim of the present study was to estimate the association between indices of laterality and reproductive indices. The studied sample comprised 650 Chuvashian women aged 18 to 80 years (mean, 46.9; SD = 16.2). The independent-sample t test was used to compare the age at menarche or age at menopause between individuals with right or left dominance of handedness, dominant eye, hand clasping, and arm folding. No significant differences in age at menarche or age at menopause between women with right and left dominance in any of the studied laterality indices were found. This is the first study that simultaneously evaluates the association between dominance in four laterality indices (handedness, dominant eye, hand clasping, and arm folding) and two reproductive indices (age at menarche and age at menopause). Result of our study do not support the hypothesis of a possible association between handedness (and other indices of laterality) and an early age at menarche or age at natural menopause.

  18. T Cell Receptor Vβ Staining Identifies the Malignant Clone in Adult T cell Leukemia and Reveals Killing of Leukemia Cells by Autologous CD8+ T cells.

    Directory of Open Access Journals (Sweden)

    Aileen G Rowan

    2016-11-01

    Full Text Available There is growing evidence that CD8+ cytotoxic T lymphocyte (CTL responses can contribute to long-term remission of many malignancies. The etiological agent of adult T-cell leukemia/lymphoma (ATL, human T lymphotropic virus type-1 (HTLV-1, contains highly immunogenic CTL epitopes, but ATL patients typically have low frequencies of cytokine-producing HTLV-1-specific CD8+ cells in the circulation. It remains unclear whether patients with ATL possess CTLs that can kill the malignant HTLV-1 infected clone. Here we used flow cytometric staining of TCRVβ and cell adhesion molecule-1 (CADM1 to identify monoclonal populations of HTLV-1-infected T cells in the peripheral blood of patients with ATL. Thus, we quantified the rate of CD8+-mediated killing of the putative malignant clone in ex vivo blood samples. We observed that CD8+ cells from ATL patients were unable to lyse autologous ATL clones when tested directly ex vivo. However, short in vitro culture restored the ability of CD8+ cells to kill ex vivo ATL clones in some donors. The capacity of CD8+ cells to lyse HTLV-1 infected cells which expressed the viral sense strand gene products was significantly enhanced after in vitro culture, and donors with an ATL clone that expressed the HTLV-1 Tax gene were most likely to make a detectable lytic CD8+ response to the ATL cells. We conclude that some patients with ATL possess functional tumour-specific CTLs which could be exploited to contribute to control of the disease.

  19. T Cell Receptor Vβ Staining Identifies the Malignant Clone in Adult T cell Leukemia and Reveals Killing of Leukemia Cells by Autologous CD8+ T cells.

    Science.gov (United States)

    Rowan, Aileen G; Witkover, Aviva; Melamed, Anat; Tanaka, Yuetsu; Cook, Lucy B M; Fields, Paul; Taylor, Graham P; Bangham, Charles R M

    2016-11-01

    There is growing evidence that CD8+ cytotoxic T lymphocyte (CTL) responses can contribute to long-term remission of many malignancies. The etiological agent of adult T-cell leukemia/lymphoma (ATL), human T lymphotropic virus type-1 (HTLV-1), contains highly immunogenic CTL epitopes, but ATL patients typically have low frequencies of cytokine-producing HTLV-1-specific CD8+ cells in the circulation. It remains unclear whether patients with ATL possess CTLs that can kill the malignant HTLV-1 infected clone. Here we used flow cytometric staining of TCRVβ and cell adhesion molecule-1 (CADM1) to identify monoclonal populations of HTLV-1-infected T cells in the peripheral blood of patients with ATL. Thus, we quantified the rate of CD8+-mediated killing of the putative malignant clone in ex vivo blood samples. We observed that CD8+ cells from ATL patients were unable to lyse autologous ATL clones when tested directly ex vivo. However, short in vitro culture restored the ability of CD8+ cells to kill ex vivo ATL clones in some donors. The capacity of CD8+ cells to lyse HTLV-1 infected cells which expressed the viral sense strand gene products was significantly enhanced after in vitro culture, and donors with an ATL clone that expressed the HTLV-1 Tax gene were most likely to make a detectable lytic CD8+ response to the ATL cells. We conclude that some patients with ATL possess functional tumour-specific CTLs which could be exploited to contribute to control of the disease.

  20. Publication point indicators

    DEFF Research Database (Denmark)

    Elleby, Anita; Ingwersen, Peter

    2010-01-01

    with novel publication point indicators (PPIs) that are formalized and exemplified. Two diachronic citation windows are applied: 2006-07 and 2006-08. Web of Science (WoS) as well as Google Scholar (GS) are applied to observe the cite delay and citedness for the different document types published by DIIS......The paper presents comparative analyses of two publication point systems, The Norwegian and the in-house system from the interdiscplinary Danish Institute for International Studies (DIIS), used as case in the study for publications published 2006, and compares central citation-based indicators......; the Cumulated Publication Point Indicator (CPPI), which graphically illustrates the cumulated gain of obtained vs. ideal points, both seen as vectors; and the normalized Cumulated Publication Point Index (nCPPI) that represents the cumulated gain of publication success as index values, either graphically...

  1. 一株裂解奶牛源致病性大肠埃希菌噬菌体的分离与鉴定%Isolation and Identification of Bacteriophage Strain Lysing Pathogenic E.coli from Cow

    Institute of Scientific and Technical Information of China (English)

    杨亚东; 王礼伟; 屈勇刚; 李飞; 杨一鸣; 杨蓓

    2014-01-01

    以实验室保存的21株致奶牛乳房炎大肠埃希菌为宿主菌,采用双层琼脂平板法从牛粪便中分离裂解性噬菌体。测定所得噬菌体的滴度、裂解谱、最佳感染复数(MOI)等生物学特性,分析其在不同 pH、温度、紫外线作用后的滴度变化,以及在-20℃条件下保存10个月的滴度变化。结果显示,分离到1株噬菌体,其滴度为7.75×1010 PFU/mL,裂解率为33.3%,最佳感染复数为10,pH 5~9范围内作用2 h、30℃~40℃时作用1 h 其滴度变化不显著,紫外线照射14 min 可被全部灭活,在-20℃条件下保存10个月仍保持活性。%Using 21 strains of pathogenic Escherichia coli isolated from cows with mastitis as host cells, bacteriophages were isolated from cow feces with double-layer agar methods.To anayze the characteristics of the bacteriophages,the titer,lysis spectrum and optimal multipicity of infection(MOI)were detected. At the same time,the thermostability,pH stability and ultraviolet ray stability were analyzed.The rusult showed that one bacteriophage was isolated and purified.The titer of the bacteriophage was 7.75 × 10 10 PFU/mL.The lysis rate was 33.3%.The MOI was 10.The bacteriophage still remained high lysis activi-ties after incubation under pH 5-9 for 2 h,or treated under 30℃-40℃ for 1 h.However,the bacteriophage could be inactivated by ultraviolet rays for 14 min.The bacteriophage remained active after storage at -20℃ for 10 months.

  2. Electrical cell counting process characterization in a microfluidic impedance cytometer.

    Science.gov (United States)

    Hassan, Umer; Bashir, Rashid

    2014-10-01

    Particle counting in microfluidic devices with coulter principle finds many applications in health and medicine. Cell enumeration using microfluidic particle counters is fast and requires small volumes of sample, and is being used for disease diagnostics in humans and animals. A complete characterization of the cell counting process is critical for accurate cell counting especially in complex systems with samples of heterogeneous population interacting with different reagents in a microfluidic device. In this paper, we have characterized the electrical cell counting process using a microfluidic impedance cytometer. Erythrocytes were lysed on-chip from whole blood and the lysing was quenched to preserve leukocytes which subsequently pass through a 15 μm × 15 μm measurement channel used to electrically count the cells. We show that cell counting over time is a non-homogeneous Poisson process and that the electrical cell counts over time show the log-normal distribution, whose skewness can be attributed to diffusion of cells in the buffer that is used to meter the blood. We further found that the heterogeneous cell population (i.e. different cell types) shows different diffusion characteristics based on the cell size. Lymphocytes spatially diffuse more as compared to granulocytes and monocytes. The time difference between the cell occurrences follows an exponential distribution and when plotted over time verifies the cell diffusion characteristics. We also characterized the probability of occurrence of more than one cell at the counter within specified time intervals using Poisson counting statistics. For high cell concentration samples, we also derived the required sample dilution based on our particle counting characterization. Buffer characterization by considering the size based particle diffusion and estimating the required dilution are critical parameters for accurate counting results.

  3. Environmental indicators for buildings

    DEFF Research Database (Denmark)

    Dammann, Sven

    2004-01-01

    Environmental Indicators for Buildings are studied using two different perspectives: with a technological, environmental scientific departing point and with a social scientific departing point. Different relevant groups in the building sector are identified and analysed, using the Social...... Construction Of Technology theory as a frame for the studies. It is concluded that it is not possible to develop a common set of indicators that all the central actors in the building sector find acceptable - at least not in a near future. Different sets of solutions are, however, outlined in the thesis...

  4. Environmental indicators for buildings

    DEFF Research Database (Denmark)

    Dammann, Sven

    Environmental Indicators for Buildings are studied using two different perspectives: with a technological, environmental scientific departing point and with a social scientific departing point. Different relevant groups in the building sector are identified and analysed, using the Social...... Construction Of Technology theory as a frame for the studies. It is concluded that it is not possible to develop a common set of indicators that all the central actors in the building sector find acceptable - at least not in a near future. Different sets of solutions are, however, outlined in the thesis...

  5. Indicações em transplante de células-tronco hematopoéticas em pacientes adultos com leucemia linfoide aguda Indications for hematopoietic stem cell transplantation in adults in acute lymphoblastic leukemia

    Directory of Open Access Journals (Sweden)

    Maria Aparecida Zanichelli

    2010-05-01

    Full Text Available Na leucemia linfoide aguda (LLA, a proliferação, acúmulo e infiltração de células imaturas caracterizam uma entidade heterogênea, apresentando ampla diversidade de aspectos clínicos e biológicos. Na LLA do adulto, a concentração de fatores prognósticos de alto risco, como o imunofenótipo B, alterações cromossômicas e, principalmente, a presença do cromossomo Ph positivo. Considerações a respeito da alta morbidade e mortalidade relacionadas ao transplante de células-tronco hematopoéticas (TCTH têm gerado controvérsias quanto à indicação desta modalidade terapêutica, nos pacientes adultos com LLA em primeira remissão (1ª RC. Os resultados da terapia convencional com quimioterapia, diante dos diferentes grupos de risco em pacientes com LLA, têm sido utilizados para a indicação de TCTH. Apresentamos o algoritmo de indicações do transplante de células-tronco hematopoéticas em pacientes adultos com LLA.In acute lymphoblastic leukemia, accumulation and proliferation of immature cells infiltration characterise a heterogeneous entity, featuring a wide variety of clinical and biological aspects. In the adult LLA concentration of high-risk prognosis factors such as age, B-cell, chromosomic changes, and chiefly the presence of chromosome positive Ph. Considerations of high morbidity and mortality rates related to haematopoietic stem cell transplantation (TCTH have generated controversy about this therapeutic modality in adult patients with LLA in first remission (1st RC. The results of conventional therapy with chemotherapy in contrast with different risk groups of patients with LLA, has been used for the indication of TCTH. Thus we present the algorithm indications of haematopoietic stem cell transplantation in adult patients with LLA. Rev.

  6. "Healthy" Human Development Indices

    Science.gov (United States)

    Engineer, Merwan; Roy, Nilanjana; Fink, Sari

    2010-01-01

    In the Human Development Index (HDI), life expectancy is the only indicator used in modeling the dimension "a long and healthy life". Whereas life expectancy is a direct measure of quantity of life, it is only an indirect measure of healthy years lived. In this paper we attempt to remedy this omission by introducing into the HDI the morbidity…

  7. Pain indicators in newborns

    Directory of Open Access Journals (Sweden)

    Spasojević Slobodan

    2008-01-01

    Full Text Available Definition of pain. The International Association for the Study of Pain has defined pain as "an unpleasant sensory and emotional experience associated with actual or potential tissue damage or described in terms of such damage." The interpretation of pain is subjective. Each person forms an internal construct of pain through encountered injury. Pain and newborn. The issue of pain perception in newborns, its management and prevention has been neglected for decades. The inability of "self-report" of painful experience has contributed significantly to misunderstanding of the importance of this problem and in­adequate treatment. The main characteristic of this 'critical window of brain development' period is rapid enlargement of brain volume and its great plasticity. Harmful short-term and long-term consequences can arise as a consequence of disturbance of the sophisticated balance between newborn and its surrounding. Neonatal pain indicators. As a response to a present painful stimulus, the newborn adapts to this acute stress with changes in endocrine, vegetative, immune and behavioral area. An ideal pain indicator in neonatal period does not exist. There are several different groups o them, namely contextual and developmental indicators (gestational age, contributed illness, medication, for example, physiological (heart rate, vagal tone, breathing rate, blood pressure, oxygen saturation, transcutaneous partial pressures of oxygen and carbon-dioxide, intracranial pressure, palm sweating and behavioral ones (face expression, movements of limbs, cry, several neonatal pain scales were constructed on the basis of these indicators. .

  8. "Healthy" Human Development Indices

    Science.gov (United States)

    Engineer, Merwan; Roy, Nilanjana; Fink, Sari

    2010-01-01

    In the Human Development Index (HDI), life expectancy is the only indicator used in modeling the dimension "a long and healthy life". Whereas life expectancy is a direct measure of quantity of life, it is only an indirect measure of healthy years lived. In this paper we attempt to remedy this omission by introducing into the HDI the morbidity…

  9. Indicators for energy security

    NARCIS (Netherlands)

    Kruyt, B.; van Vuuren, D.P.; de Vries, H.J.M.; Groenenberg, H.

    2009-01-01

    The concept of energy security is widely used, yet there is no consensus on its precise interpretation. In this research, we have provided an overview of available indicators for long-term security of supply (SOS). We distinguished four dimensions of energy security that relate to the availability,

  10. Publication point indicators

    DEFF Research Database (Denmark)

    Elleby, Anita; Ingwersen, Peter

    2010-01-01

    with novel publication point indicators (PPIs) that are formalized and exemplified. Two diachronic citation windows are applied: 2006-07 and 2006-08. Web of Science (WoS) as well as Google Scholar (GS) are applied to observe the cite delay and citedness for the different document types published by DIIS...

  11. Maslov indices and monodromy

    NARCIS (Netherlands)

    Dullin, H.R.; Robbins, J.M.; Waalkens, H.; Creagh, S.C.; Tanner, G.

    2005-01-01

    We prove that for a Hamiltonian system on a cotangent bundle that is Liouville-integrable and has monodromy the vector of Maslov indices is an eigenvector of the monodromy matrix with eigenvalue 1. As a corollary, the resulting restrictions on the monodromy matrix are derived.

  12. Environmental health indicators

    Directory of Open Access Journals (Sweden)

    Manuel Posada de la Paz

    2004-12-01

    Full Text Available This presentation gives a general overview of the project titled Environmental Health Indicators, coordinated by WHO and managed by the Research Centre on the Toxic Oil Syndrome and Rare Diseases in Spain. The presentations describes the objectives of the project, the steps taken and the results obtained during the feasibility study. The project aims to develop an environmental health information system that will allow the monitoring of public health and its health determinants and make international comparisons, develop environmental policies and facilitate communication with citizens. WHO developed a methodology for the development of these indicators within the conceptual framework of DPSEEA (Driving Force, Pressure, State, Exposure, Effect, Action and selected a total of 55 indicators (which included 168 variables in 10 environmental health areas. The feasibility study predicted the successful gathering of 89% of the indicators. However, data recollection proved difficult due to the frequent incompatibility of some variables in the Spanish information systems with the WHO defined variables. On a management level, the greatest difficulty arose from the disperse distribution of responsibilities in environmental health matters. In addition to the technical contribution of this project to Environmental Health in Spain, an added value has been to establish a close collaboration with the different Ministries involved.

  13. The logarithmic hypervolume indicator

    DEFF Research Database (Denmark)

    Friedrich, Tobias; Bringmann, Karl; Voß, Thomas;

    2011-01-01

    It was recently proven that sets of points maximizing the hypervolume indicator do not give a good multiplicative approximation of the Pareto front. We introduce a new “logarithmic hypervolume indicator” and prove that it achieves a close-to-optimal multiplicative approximation ratio. This is exp...

  14. Effect of chronic alcohol consumption on the red blood cell count and RBC indices in the HIV infected patients on d4T/3TC/NVP drug regimen in Uganda

    Directory of Open Access Journals (Sweden)

    Godfrey S. Bbosa

    2013-10-01

    Full Text Available Alcohol consumption is common problem in Uganda. Among the types of alcohols consumed include beers, spirits, liqueurs, wines and traditional brew. These alcohols are easily accessible and consumed by many people including the HIV infected patients who are on the d4T/3TC/NVP regimen. The aim of this study was to determine the effect of chronic alcohol intake on the red blood cell count (RBC and the RBC indices in the HIV-infected patients on d4T/3TC/NVP regimen. It was a case control study that used a repeated measures design model where serial measurements of the red blood cell count (RBC and RBC indices were determined at 3 month interval for 9 months. A total of 41 HIV infected patients were recruited and grouped into two arms; the control group had 21 patients and the chronic alcohol group had 20 patients. The RBC and RBC indices of the whole blood were determined using automated hematological Coulter CBC-5 Hematology Analyzer system using standard procedures. The data was sorted into alcohol-use self reporting by WHO AUDIT tool and alcohol-use biomarkers groups. It was analysed using the SAS 2003 version 9.1 statistical package with the repeated measures fixed model. The means were compared using the student t-test. The mean MCV and MCH values in the chronic alcohol use group were higher than in the control group and there was a significant difference between the 2 groups (p<0.05 for both the WHO AUDIT tool group and chronic alcohol use biomarkers group. The mean RBC count, Hct, HGB and MCHC values in both the control and chronic alcohol use groups were within the normal reference ranges for both groups though the trend was lower in alcohol group. Chronic alcohol use affects the RBC and RBC indices in the HIV infected patients on d4T/3TC/NVP treatment regimen. [Int J Basic Clin Pharmacol 2013; 2(5.000: 528-536

  15. 波纹唇鱼血细胞显微结构和血液生化指标%Microstructure of Peripheral Blood Cell and Serum Biochemical Indices of Cheilinus undulatus

    Institute of Scientific and Technical Information of China (English)

    廖光勇; 区又君; 李加儿

    2011-01-01

    以Wright's和Giemsa双重染色法对波纹唇鱼(Cheilinus undulatus)外周血细胞的形态进行显微观察研究并计数,同时测定各类血细胞的大小及血液的生化指标.血涂片观察发现:波纹唇鱼的外周血细胞可区分出红细胞、嗜中性粒细胞、单核细胞、淋巴细胞、血栓细胞等五种血细胞,未观察到嗜碱性粒细胞、嗜酸性粒细胞.在外周血液中还观察到核影、正在分裂的红细胞、幼稚红细胞和降解的红细胞.各类血细胞胞体由大到小的次序依次为单核细胞、嗜中性粒细胞、红细胞、大淋巴细胞、血栓细胞、小淋巴细胞.血浆中红细胞为(3.15±0.79)×1012/L,白细胞为(1.42±0.71)×1010/L,红细胞数远远多于白细胞.血液生化指标与其他鱼类相比有一定差异,表明鱼类血液的生化成分因种和生活环境而异.%The microscopic observation and count are conducted for the peripheral blood cells of C. Undula-tus by means of the Wright's and Giemsa double dye method. The serum biochemical indices of C. Undula-tus , the sizes of the various blood cells are determined. From the stained smears, five blood cell types I. E. Erythrocyte, neutrophil, monocyte, lymphocyte and thrombocyte are detected. Neither eosinophilic granu-locyten nor basophilic granulocyte is found but, some nuclear shadow of erythrocytes, immature erythro-cytes, direct dividing or dissolving erythrocytes. Among these cells, the sizes are put into an order from large to small as follows monocyte, neutrophil, erythrocyte, large lymphocyte, thrombocyte and small lymphlcyte. The erythrocyte and leucocyte concentrations in the blood are (3. 15 ±0. 79) × 1012/L and (1. 42 + 0. 71) × 1010/L respectively. The number of erythrocyte is much more than the leucocyte in the blood. The serum biochemical indices are different from other fish species. Therefore it is indicated that the serum biochemical indices for various fishes depend upon their species and

  16. GRB as luminosity indicator

    CERN Document Server

    Basak, Rupal

    2014-01-01

    Gamma Ray Bursts (GRBs) are found at much higher redshifts (z>6) than Supernova Ia (z~1), and hence, they can be used to probe very primitive universe. However, radiation mechanism of GRB remains a puzzle, unlike Supernova Ia. Through comprehensive description, both empirical and physical, we shall discuss the most likely way to use the constituent pulses of a GRB to find the radiation mechanism as well as using the pulses as luminosity indicators.

  17. Indicators for Knowledge Economy

    OpenAIRE

    2007-01-01

    The Lisbon European Council conclusion was that in 2010 Europe will become 'the most competitive and dynamic knowledge-based economy in the world, capable of sustained economic growth with more and better jobs and greater social cohesion'. The knowledge economy concept is a part of modern society. This paper examines the knowledge economy concept and indicators for measuring the performance of the knowledge economy

  18. Indicators of Ecological Change

    Science.gov (United States)

    2005-03-03

    Wang et al. (2003) and earlier those of Chapra and Di Toro (1991). Wang et al. (2003) termed their approach the extreme value method and used it to...Thesis, Auburn University, Auburn, AL. 210 pp. Chapra , S. C., Di Toro, D. M., 1991. Delta method for estimating primary production, respiration, and...characteristics, The challenge in using ecological indicators is determining which of the numerous measures of ecological systems best characterize

  19. Simplicial Indicator Kriging

    Institute of Scientific and Technical Information of China (English)

    R Tolosana-Delgado; V Pawlowsky-Glahn; J J Egozcue

    2008-01-01

    Indicator kriging (IK) is a spatial interpolation technique devised for estimating a conditional cumulative distribution function at an unsampled location. The result is a discrete approximation, and its corresponding estimated probability density function can be viewed as a composition in the simplex. This fact suggested a compositional approach to IK which, by construction, avoids all its standard drawbacks (negative predictions, not-ordered or larger than one). Here, a simple algorithm to develop the procedure is presented.

  20. Indicators of Ecological Change

    Science.gov (United States)

    2005-03-01

    measures Leaf area Branch evaluations Plant diversity Ozone bioindicators Lichen diversity Visible damage Vegetation structure Forest type...composition, and physiological status (measured as signature lipid/ DNA biomarkers) as a quick and easy indicator of changes in soil quality as a...Cryptophyte Geophyte Forb 0.17 0.45 Blazing star RL Liatris tenuafolia Asteraceae Cryptophyte Geophyte Forb 0.11 0.36 Blazing star RM Lichens Lichens 0.61

  1. Quantification of damage due to low-dose radiation exposure in mice: construction and application of a biodosimetric model using mRNA indicators in circulating white blood cells.

    Science.gov (United States)

    Ishihara, Hiroshi; Tanaka, Izumi; Yakumaru, Haruko; Tanaka, Mika; Yokochi, Kazuko; Fukutsu, Kumiko; Tajima, Katsushi; Nishimura, Mayumi; Shimada, Yoshiya; Akashi, Makoto

    2016-01-01

    Biodosimetry, the measurement of radiation damage in a biologic sample, is a reliable tool for increasing the accuracy of dose estimation. Although established chromosome analyses are suitable for estimating the absorbed dose after high-dose irradiation, biodosimetric methodology to measure damage following low-dose exposure is underdeveloped. RNA analysis of circulating blood containing radiation-sensitive cells is a candidate biodosimetry method. Here we quantified RNA from a small amount of blood isolated from mice following low-dose body irradiation (radiation-sensitive undifferentiated cells in the blood based on Myc RNA expression, we quantified the relative levels of RNA for DNA damage-induced (DDI) genes, such as Bax, Bbc3 and Cdkn1a. The RNA ratios of DDI genes/Myc in the blood increased in a dose-dependent manner 4 h after whole-body irradiation at doses ranging from 0.1 to 0.5 Gy (air-kerma) of X-rays, regardless of whether the mice were in an active or resting state. The RNA ratios were significantly increased after 0.014 Gy (air-kerma) of single X-ray irradiation. The RNA ratios were directly proportional to the absorbed doses in water ranging from 0.1 to 0.5 Gy, based on gamma-irradiation from (137)Cs. Four hours after continuous irradiation with gamma-rays or by internal contamination with a beta-emitter, the increased RNA ratios resembled those following single irradiation. These findings indicate that the RNA status can be utilized as a biodosimetric tool to estimate low-dose radiation when focusing on undifferentiated cells in blood. © The Author 2015. Published by Oxford University Press on behalf of The Japan Radiation Research Society and Japanese Society for Radiation Oncology.

  2. Human cell exposure assays of Bacillus thuringiensis commercial insecticides: production of Bacillus cereus-like cytolytic effects from outgrowth of spores.

    Science.gov (United States)

    Tayabali, A F; Seligy, V L

    2000-01-01

    Most contemporary bioinsecticides are derived from scaled-up cultures of Bacillus thuringiensis subspecies israelensis (Bti) and kurstaki (Btk), whose particulate fractions contain mostly B. thuringiensis spores (> 10(12)/L) and proteinaceous aggregates, including crystal-like parasporal inclusion bodies (PIB). Based on concerns over relatedness to B. cereus-group pathogens, we conducted extensive testing of B. thuringiensis (BT) products and their subfractions using seven human cell types. The Bti/Btk products generated nonspecific cytotoxicities involving loss in bioreduction, cell rounding, blebbing and detachment, degradation of immunodetectable proteins, and cytolysis. Their threshold dose (Dt approximately equal.5 times 10(-14)% BT product/target cell) equated to a single spore and a target cell half-life (tLD(50)) of approximately 16 hr. At Dts > 10(4), the tLD(50) rapidly shifted to < 4 hr; with antibiotic present, no component, including PIB-related [delta]-endotoxins, was cytolytic up to an equivalent of approximately 10(9 )Dt. The cytolytic agent(s) within the Bti/Btk-vegetative cell exoprotein (VCP) pool is an early spore outgrowth product identical to that of B. cereus and acting possibly by arresting protein synthesis. No cytolytic effects were seen with VCP from B. subtilis and Escherichia coli. These data, including recent epidemiologic work indicate that spore-containing BT products have an inherent capacity to lyse human cells in free and interactive forms and may also act as immune sensitizers. To critically impact at the whole body level, the exposure outcome would have to be an uncontrolled infection arising from intake of Btk/Bti spores. For humans, such a condition would be rare, arising possibly in equally rare exposure scenarios involving large doses of spores and individuals with weak or impaired microbe-clearance capacities and/or immune response systems. PMID:11049810

  3. Isolation of dimorphic chloroplasts from the single-cell C4 species Bienertia sinuspersici

    Directory of Open Access Journals (Sweden)

    Lung Shiu-Cheung

    2012-03-01

    Full Text Available Abstract Three terrestrial plants are known to perform C4 photosynthesis without the dual-cell system by partitioning two distinct types of chloroplasts in separate cytoplasmic compartments. We report herein a protocol for isolating the dimorphic chloroplasts from Bienertia sinuspersici. Hypo-osmotically lysed protoplasts under our defined conditions released intact compartments containing the central chloroplasts and intact vacuoles with adhering peripheral chloroplasts. Following Percoll step gradient purification both chloroplast preparations demonstrated high homogeneities as evaluated from the relative abundance of respective protein markers. This protocol will open novel research directions toward understanding the mechanism of single-cell C4 photosynthesis.

  4. Attachment of an anti-receptor antibody to non-target cells renders them susceptible to lysis by a clone of cytotoxic T lymphocytes.

    Science.gov (United States)

    Kranz, D M; Tonegawa, S; Eisen, H N

    1984-12-01

    The molecular basis for the dependence of antigen recognition by T cells on products of the major histocompatibility complex (MHC) is unknown, and the antigenic structures that are actually bound by T-cell receptors are ill-defined. In this study, we asked whether a monoclonal antibody (mAb) that reacts with the T-cell receptor of a clone of murine cytotoxic T lymphocytes (CTL) and not with the receptors of other CTL clones can substitute for that clone's natural ligand in specific cytolytic reactions. To answer the question, a mAb (1B2) to the receptor of a CTL clone (2C) was attached covalently to 51Cr-labeled cells that were not otherwise susceptible to lysis by clone 2C, and the cells thus modified were then tested as targets for clone 2C and other CTL clones of similar specificity. All labeled cells modified in this way, including a murine cell line that expresses no cell-surface MHC class I molecules and a human cell line, were lysed by clone 2C but not by other CTL clones. If, however, instead of attaching the mAb to the receptor of clone 2C, the cells were modified by attaching to them mAbs to other surface antigens on CTL [lymphocyte function-associated antigen (LFA-1), Thy-1.2], they were not lysed. In cytolytic titrations, the cells that had been converted by attachment of mAb 1B2 into specific targets for clone 2C were just as susceptible to lysis by that clone as the clone's natural H-2d targets (e.g., P815 cells). However, some accessory surface molecules (LFA-1, Lyt-2) that are required for clone 2C to lyse its natural H-2d targets seemed not to be required for this clone to lyse the mAb-converted target cells. By demonstrating that a variety of different cell types can be thus converted into target cells for CTL, the approach described in this study may provide opportunities to analyze further the mechanisms by which CTL destroy target cells.

  5. Myristoylation profiling in human cells and zebrafish

    Directory of Open Access Journals (Sweden)

    Malgorzata Broncel

    2015-09-01

    Full Text Available Human cells (HEK 293, HeLa, MCF-7 and zebrafish embryos were metabolically tagged with an alkynyl myristic acid probe, lysed with an SDS buffer and tagged proteomes ligated to multifunctional capture reagents via copper-catalyzed alkyne azide cycloaddition (CuAAC. This allowed for affinity enrichment and high-confidence identification, by delivering direct MS/MS evidence for the modification site, of 87 and 61 co-translationally myristoylated proteins in human cells and zebrafish, respectively. The data have been deposited to ProteomeXchange Consortium (Vizcaíno et al., 2014 Nat. Biotechnol., 32, 223–6 (PXD001863 and PXD001876 and are described in detail in Multifunctional reagents for quantitative proteome-wide analysis of protein modification in human cells and dynamic protein lipidation during vertebrate development׳ by Broncel et al., Angew. Chem. Int. Ed.

  6. Mitochondrial bioelectrocatalysis for biofuel cell applications

    Energy Technology Data Exchange (ETDEWEB)

    Arechederra, Robert L.; Boehm, Kevin [Saint Louis University, Department of Chemistry, 3501 Laclede Ave., St. Louis, MO 63103 (United States); Minteer, Shelley D., E-mail: minteers@slu.ed [Saint Louis University, Department of Chemistry, 3501 Laclede Ave., St. Louis, MO 63103 (United States)

    2009-12-01

    Mitochondria modified electrodes have been developed and characterized that utilize whole mitochondria isolated from tubers and immobilized within a quaternary ammonium modified Nafion membrane on a carbon electrode that can oxidize pyruvate and fatty acids. Detailed characterization of the performance of these mitochondria modified electrodes has been accomplished by coupling the mitochondria-based bioanode with a commercial air breathing cathode in a complete pyruvate/air biofuel cell. The studies included the effect of fuel (pyruvate) concentration, mitochondria lysing, temperature and pH on the performance of the mitochondria catalyzed, pyruvate/air biofuel cell. Effect of oxygen and cytochrome c oxidase inhibitors on biofuel cell performance has allowed us to further understand the mechanism of electron transfer with the carbon electrode.

  7. Rapid thermal lysis of cells using silicon-diamond microcantilever heaters.

    Science.gov (United States)

    Privorotskaya, Natalya; Liu, Yi-Shao; Lee, Jungchul; Zeng, Hongjun; Carlisle, John A; Radadia, Adarsh; Millet, Larry; Bashir, Rashid; King, William P

    2010-05-07

    This paper presents the design and application of microcantilever heaters for biochemical applications. Thermal lysis of biological cells was demonstrated as a specific example. The microcantilever heaters, fabricated from selectively doped single crystal silicon, provide local resistive heating with highly uniform temperature distribution across the cantilevers. Very importantly, the microcantilever heaters were coated with a layer of 100 nm thick electrically insulating ultrananocrystalline diamond (UNCD) layer used for cell immobilization on the cantilever surface. Fibroblast cells or bacterial cells were immobilized on the UNCD/cantilever surfaces and thermal lysis was demonstrated via optical fluorescence microscopy. Upon electrical heating of the cantilever structures to 93 degrees C for 30 seconds, fibroblast cell and nuclear membrane were compromised and the cells were lysed. Over 90% of viable bacteria were also lysed after 15 seconds of heating at 93 degrees C. This work demonstrates the utility of silicon-UNCD heated microcantilevers for rapid cell lysis and forms the basis for other rapid and localized temperature-regulated microbiological experiments in cantilever-based lab on chip applications.

  8. A novel, compact disk-like centrifugal microfluidics system for cell lysis and sample homogenization.

    Science.gov (United States)

    Kido, Horacio; Micic, Miodrag; Smith, David; Zoval, Jim; Norton, Jim; Madou, Marc

    2007-07-01

    In this paper, we present the design and characterization of a novel platform for mechanical cell lysis of even the most difficult to lyse cell types on a micro or nanoscale (maximum 70 microL total volume). The system incorporates a machined plastic circular disk assembly, magnetic field actuated microfluidics, centrifugal cells and tissue homogenizer and centrifugation system. The mechanism of tissue disruption of this novel cell homogenization apparatus derives from the relative motion of ferromagnetic metal disks and grinding matrices in a liquid medium within individual chambers of the disk in the presence of an oscillating magnetic field. The oscillation of the ferromagnetic disks or blades produces mechanical impaction and shear forces capable of disrupting cells within the chamber both by direct action of the blade and by the motion of the surrounding lysis matrix, and by motion induced vortexing of buffer fluid. Glass beads or other grinding media are integrated into each lysis chamber within the disk to enhance the transfer of energy from the oscillating metal blade to the cells. The system also achieves the centrifugal elimination of solids from each liquid sample and allows the elution of clarified supernatants via siphoning into a collection chamber fabricated into the plastic disk assembly. This article describes system design, implementation and validation of proof of concept on two samples--Escherichia coli and Saccharomyces cerevisiae representing model systems for cells that are easy and difficult to lyse, respectively.

  9. Teriparatide - Indications beyond osteoporosis

    Directory of Open Access Journals (Sweden)

    Marilyn Lee Cheng

    2012-01-01

    Full Text Available Osteoporosis is a condition of impaired bone strength that results in an increased risk of fracture. The current and most popular pharmacological options for the treatment of osteoporosis include antiresorptive therapy, in particular, oral bisphosphonates (alendronate, risedronate, ibandronate. Anabolic agents like teriparatide have widened our therapeutic options. They act by directly stimulating bone formation and improving bone mass quantity and quality. Two forms of recombinant human parathyroid hormone (PTH are available : full-length PTH (PTH 1-84; approved in the EU only and the 1-34 N-terminal active fragment of PTH (teriparatide, US FDA approved. This review aims to discuss the benefits of teriparatide beyond the currently licensed indications like fracture healing, dental stability, osteonecrosis of jaw, hypoparathyroidism, and hypocalcemia.

  10. INDICATORS OF TERRITORIAL COHESION

    Directory of Open Access Journals (Sweden)

    MARIA VASILESCU (DUMITRASCU

    2014-12-01

    Full Text Available In a world of globalization generating geographical concentration, relocation of activities, thus inducing territorial disparities, territorial cohesion has become a prerequisite for achieving sustainable economic growth and implementing social and economic cohesion. The ratification of Lisbon Treaty represents the moment when “Territorial Cohesion” has been added as an official policy aim of the EU to its previous aims of economic and social cohesion. Thus, the process of defining this concept of “Territorial Cohesion” has become the task of the Commission and EU Member States. Based on a literature review, the paper describes, in the first section, the polysemantic nature of territorial cohesion. The main objective of the next section is to address the main indicators, directly related to territorial objectives, that could be used to support policy makers in measuring and monitoring territorial cohesion related to European territorial development.

  11. Environmental indicators for buildings

    DEFF Research Database (Denmark)

    Dammann, S.

    Whenever we shop, the products we consider buying are labelled with the economical price we have to pay if we want to purchase them - an important parameter in our decisions as purchasers. The increrasing awareness for environmental limits and backlashes of human activities also in the building...... sector have fostered the wish to define 'the ecological price' of a building as a help for environmental conscious decision-making. In a social constructivist approach this Ph.D. thesis looks across and beyond the manifold existing approaches for environmental indicators for buildings. It acknowledges...... that among the relevant actors in the building sector the scientific view is only one perspective among others. This study combines natural-scientific knowledge with social-scientific knowledge, obtained in a close co-operation with actors in the building sector in Denmark and a research period...

  12. Modelling T4 cell count as a marker of HIV progression in the absence of any defence mechanism

    Directory of Open Access Journals (Sweden)

    VSS Yadavalli

    2010-12-01

    Full Text Available The T4 cell count, which is considered one of the markers of disease progression in an HIV infected individual, is modelled in this paper. The World Health Organisation has recently advocated that countries encourage HIV infected individuals to commence antiretroviral treatments once their T4 cell count drops below 350 cells per ml of blood (this threshold was formerly 200 cells per ml of blood. This recommendation is made because when the T4 cell count is low, the T4 cells are unable to mount an effective immune response against antigens and any such foreign matters in the body, and consequently the individual becomes susceptible to opportunistic infections and lymphomas. A stochastic catastrophe model is developed in this paper to obtain the mean, variance and covariance of the uninfected, infected and lysed T4 cells. The amount of toxin produced in an HIV infected person from the time of infection to a later time may also be obtained from the model. Numerical illustrations of the correlation structures between uninfected and infected T4 cells, and between the infected and lysed T4 cells are also presented.

  13. Isolation of highly suppressive CD25+FoxP3+ T regulatory cells from G-CSF-mobilized donors with retention of cytotoxic anti-viral CTLs: application for multi-functional immunotherapy post stem cell transplantation.

    Science.gov (United States)

    Samuel, Edward R; Beloki, Lorea; Newton, Katy; Mackinnon, Stephen; Lowdell, Mark W

    2014-01-01

    Previous studies have demonstrated the effective control of cytomegalovirus (CMV) infections post haematopoietic stem cell transplant through the adoptive transfer of donor derived CMV-specific T cells (CMV-T). Strategies for manufacturing CMV immunotherapies has involved a second leukapheresis or blood draw from the donor, which in the unrelated donor setting is not always possible. We have investigated the feasibility of using an aliquot of the original G-CSF-mobilized graft as a starting material for manufacture of CMV-T and examined the activation marker CD25 as a targeted approach for identification and isolation following CMVpp65 peptide stimulation. CD25+ cells isolated from G-CSF-mobilized apheresis revealed a significant increase in the proportion of FoxP3 expression when compared with conventional non-mobilized CD25+ cells and showed a superior suppressive capacity in a T cell proliferation assay, demonstrating the emergence of a population of Tregs not present in non-mobilized apheresis collections. The expansion of CD25+ CMV-T in short-term culture resulted in a mixed population of CD4+ and CD8+ T cells with CMV-specificity that secreted cytotoxic effector molecules and lysed CMVpp65 peptide-loaded phytohaemagglutinin-stimulated blasts. Furthermore CD25 expanded cells retained their suppressive capacity but did not maintain FoxP3 expression or secrete IL-10. In summary our data indicates that CD25 enrichment post CMV stimulation in G-CSF-mobilized PBMCs results in the simultaneous generation of both a functional population of anti-viral T cells and Tregs thus illustrating a potential single therapeutic strategy for the treatment of both GvHD and CMV reactivation following allogeneic haematopoietic stem cell transplantation. The use of G-CSF-mobilized cells as a starting material for cell therapy manufacture represents a feasible approach to alleviating the many problems incurred with successive donations and procurement of cells from unrelated donors

  14. CD19-CAR engineered NK-92 cells are sufficient to overcome NK cell resistance in B-cell malignancies.

    Science.gov (United States)

    Romanski, Annette; Uherek, Christoph; Bug, Gesine; Seifried, Erhard; Klingemann, Hans; Wels, Winfried S; Ottmann, Oliver G; Tonn, Torsten

    2016-07-01

    Many B-cell acute and chronic leukaemias tend to be resistant to killing by natural killer (NK) cells. The introduction of chimeric antigen receptors (CAR) into T cells or NK cells could potentially overcome this resistance. Here, we extend our previous observations on the resistance of malignant lymphoblasts to NK-92 cells, a continuously growing NK cell line, showing that anti-CD19-CAR (αCD19-CAR) engineered NK-92 cells can regain significant cytotoxicity against CD19 positive leukaemic cell lines and primary leukaemia cells that are resistant to cytolytic activity of parental NK-92 cells. The 'first generation' CAR was generated from a scFv (CD19) antibody fragment, coupled to a flexible hinge region, the CD3ζ chain and a Myc-tag and cloned into a retrovirus backbone. No difference in cytotoxic activity of NK-92 and transduced αCD19-CAR NK-92 cells towards CD19 negative targets was found. However, αCD19-CAR NK-92 cells specifically and efficiently lysed CD19 expressing B-precursor leukaemia cell lines as well as lymphoblasts from leukaemia patients. Since NK-92 cells can be easily expanded to clinical grade numbers under current Good Manufactoring Practice (cGMP) conditions and its safety has been documented in several phase I clinical studies, treatment with CAR modified NK-92 should be considered a treatment option for patients with lymphoid malignancies.

  15. Dynamic Contrast-Enhanced Magnetic Resonance Imaging Rapidly Indicates Vessel Regression in Human Squamous Cell Carcinomas Grown in Nude Mice Caused by VEGF Receptor 2 Blockade with DC101

    Directory of Open Access Journals (Sweden)

    Fabian Kiessling

    2004-05-01

    Full Text Available The purpose of our study was the investigation of early changes in tumor vascularization during antiangiogenic therapy with the vascular endothelial growth factor (VEGF receptor 2 antibody (DC101 using dynamic contrast-enhanced magnetic resonance imaging (DCE MRI. Subcutaneous heterotransplants of human skin squamous cell carcinomas in nude mice were treated with DC101. Animals were examined before and repeatedly during 2 weeks of antiangiogenic treatment using Gd-DTPA-enhanced dynamic T1-weighted MRI. With a two-compartment model, dynamic data were parameterized in "amplitude" (increase of signal intensity relative to precontrast value and kep (exchange rate constant. Data obtained by MRI were validated by parallel examinations of histological sections immunostained for blood vessels (CD31. Already 2 days after the first DC101 application, a decrease of tumor vascularization was observed, which preceded a reduction of tumor volume. The difference between treated tumors and controls became prominent after 4 days, when amplitudes of treated tumors were decreased by 61% (P = .02. In line with change of microvessel density, the decrease in amplitudes was most pronounced in tumor centers. On day 7, the mean tumor volumes of treated (153 ± 843 mm3 and control animals (596 ± 384 mm3 were significantly different (P = .03. After 14 days, treated tumors showed further growth reduction (83 ± 93 mm3, whereas untreated tumors (1208±822 mm3 continued to increase (P=.02. Our data underline the efficacy of DC101 as antiangiogenic treatment in human squamous cell carcinoma xenografts in nude mice and indicate DCE MRI as a valuable tool for early detection of treatment effects before changes in tumor volume become apparent.

  16. Possible role of minor h antigens in the persistence of donor chimerism after stem cell transplantation; relevance for sustained leukemia remission.

    Directory of Open Access Journals (Sweden)

    Cornelis R van der Torren

    Full Text Available Persistent complete donor chimerism is an important clinical indicator for remissions of hematological malignancies after HLA-matched allogeneic stem cell transplantation (SCT. However, the mechanisms mediating the persistence of complete donor chimerism are poorly understood. The frequent coincidence of complete donor chimerism with graft-versus-leukemia effects and graft-versus-host disease suggests that immune responses against minor histocompatibility antigens (mHags are playing an important role in suppressing the host hematopoiesis after allogeneic SCT. Here, we investigated a possible relationship between donor immune responses against the hematopoiesis-restricted mHag HA-1 and the long-term kinetics of host hematopoietic chimerism in a cohort of 10 patients after allogeneic HLA-matched, HA-1 mismatched SCT. Functional HA-1 specific CTLs (HA-1 CTLs were detectable in 6/10 patients lysing host-type hematopoietic cells in vitro. Presence of HA-1 CTLs in the peripheral blood coincided with low host hematopoiesis levels quantified by highly sensitive mHag specific PCR. Additionally, co-incubation of host type CD34+ cells with HA-1 CTLs isolated after allogeneic SCT prevented progenitor and cobblestone area forming cell growth in vitro and human hematopoietic engraftment in immunodeficient mice. Conversely, absence or loss of HA-1 CTLs mostly coincided with high host hematopoiesis levels and/or relapse. In summary, in this first study, presence of HA-1 CTLs paralleled low host hematopoiesis levels. This coincidence might be supported by the capacity of HA-1 CTLs isolated after allogeneic SCT to specifically eliminate host type hematopoietic stem/progenitor cells. Additional studies involving multiple mismatched mHags in more patients are required to confirm this novel characteristic of mHag CTLs as factor for the persistence of complete donor chimerism and leukemia remission after allogeneic SCT.

  17. Entanglement–breaking indices

    Energy Technology Data Exchange (ETDEWEB)

    Lami, L. [Scuola Normale Superiore, I-56126 Pisa (Italy); Giovannetti, V. [NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, I-56127 Pisa (Italy)

    2015-09-15

    We study a set of new functionals (called entanglement–breaking indices) which characterize how many local iterations of a given (local) quantum channel are needed in order to completely destroy the entanglement between the system of interest over which the transformation is defined and an external ancilla. The possibility of contrasting the noisy effects introduced by the channel iterations via the action of intermediate (filtering) transformations is analyzed. We provide some examples in which our functionals can be exactly calculated. The differences between unitary and non-unitary filtering operations are analyzed showing that, at least for systems of dimension d larger than or equal to 3, the non-unitary choice is preferable (the gap between the performances of the two cases being divergent in some cases). For d = 2 (qubit case), on the contrary, no evidences of the presence of such gap is revealed: we conjecture that for this special case unitary filtering transformations are optimal. The scenario in which more general filtering protocols are allowed is also discussed in some detail. The case of a depolarizing noise acting on a two–qubit system is exactly solved in a general case.

  18. Thaw-and-use target cells pre-labeled with calcein AM for antibody-dependent cell-mediated cytotoxicity assays.

    Science.gov (United States)

    Chung, Shan; Nguyen, Van; Lin, Yuwen Linda; Kamen, Lynn; Song, An

    2017-08-01

    In vitro antibody-dependent cell-mediated cytotoxicity (ADCC) assays are routinely performed to support the research and development of therapeutic antibodies. In ADCC assays, target cells bound by the antibodies are lysed by activated effector cells following interactions between the Fc region of the bound antibody and Fcγ receptors on effector cells. Target cell lysis is typically measured by quantification of released endogenous enzymes, e.g., lactate dehydrogenase, or measurement of released exogenous labels, e.g., (51)Cr, europium or calcein. ADCC assays based on the detection of exogenous labels released from lysed target cells generally show higher sensitivity and require shorter incubation times. However, target cells are usually labeled immediately prior to assay, which inadvertently introduces additional assay variations due to differences in target cell conditions and labeling/handling processes. In this report, we describe the use of thaw-and-use pre-labeled target cells for ADCC assays. Thaw-and-use target cells in our experiments were pre-labeled with the fluorescent dye calcein AM, cryopreserved in single-use aliquots and used directly in assays after thawing. Upon thaw, the pre-labeled cells displayed viability and label retention comparable to freshly labeled cells, responded to ADCC mediated by both peripheral blood mononuclear cells and engineered natural killer cells, performed stably for at least 3 years and provided favorable precision and accuracy to ADCC assays. Implementation of thaw-and-use pre-labeled target cells in ADCC assays can help to alleviate both cell culture and dye labeling derived variability, increase the flexibility of assay scheduling and improve assay consistency and robustness. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Microbes as forensic indicators.

    Science.gov (United States)

    Alan, G; Sarah, J P

    2012-09-01

    The forensic potential of microorganisms is becoming increasingly apparent as a consequence of advances in molecular sciences and genomics. This review discusses instances in which microbes, and in particular bacteria, can impact upon forensic investigations. There is increasing evidence that humans have an extremely diverse 'microbiome' that may prove useful in determining ethnicity, country of origin, and even personal identity. The human microbiome differs between regions of the body and may prove useful for determining the nature of stains such as those caused by saliva and vaginal fluid: it may even be possible to link the stains to the person responsible for them. Similarly, the composition of the microbiome present in a soil sample may prove a useful indicator of geographic origin or as a means of linking people, animals, or objects together or to a specific location. Microorganisms are important in the decay process and also influence the presence and concentration of alcohol, drugs, and other chemicals of forensic relevance. There is also a possibility that the entry of microorganisms into the body during the agonal period may prove useful for the diagnosis of drowning. The transmission of infectious diseases, and in particular sexually-transmitted diseases, can provide evidence linking a victim and a suspect. Microorganisms that cause fatal infections are not always identified at the time of death and may lead to the death being considered 'suspicious'. If a fatal infection can be linked to a hospital or medical procedure it can lead to prosecutions and therefore it is important to determine when and where an infection was acquired. Similarly, naturally acquired infections need to be distinguished from those that result from malicious transmission. Microorganisms can therefore provide evidence in many different forensic scenarios but most of the work is still at the experimental stage and there are therefore many opportunities for further research.

  20. Solar Indices Forecasting Tool

    Science.gov (United States)

    Henney, Carl John; Shurkin, Kathleen; Arge, Charles; Hill, Frank

    2016-05-01

    Progress to forecast key space weather parameters using SIFT (Solar Indices Forecasting Tool) with the ADAPT (Air Force Data Assimilative Photospheric flux Transport) model is highlighted in this presentation. Using a magnetic flux transport model, ADAPT, we estimate the solar near-side field distribution that is used as input into empirical models for predicting F10.7(solar 10.7 cm, 2.8 GHz, radio flux), the Mg II core-to-wing ratio, and selected bands of solar far ultraviolet (FUV) and extreme ultraviolet (EUV) irradiance. Input to the ADAPT model includes the inferred photospheric magnetic field from the NISP ground-based instruments, GONG & VSM. Besides a status update regarding ADAPT and SIFT models, we will summarize the findings that: 1) the sum of the absolute value of strong magnetic fields, associated with sunspots, is shown to correlate well with the observed daily F10.7 variability (Henney et al. 2012); and 2) the sum of the absolute value of weak magnetic fields, associated with plage regions, is shown to correlate well with EUV and FUV irradiance variability (Henney et al. 2015). This work utilizes data produced collaboratively between Air Force Research Laboratory (AFRL) and the National Solar Observatory (NSO). The ADAPT model development is supported by AFRL. The input data utilized by ADAPT is obtained by NISP (NSO Integrated Synoptic Program). NSO is operated by the Association of Universities for Research in Astronomy (AURA), Inc., under a cooperative agreement with the National Science Foundation (NSF). The 10.7 cm solar radio flux data service, utilized by the ADAPT/SIFT F10.7 forecasting model, is operated by the National Research Council of Canada and National Resources Canada, with the support of the Canadian Space Agency.

  1. Electrolyte leakage as an indicator

    Directory of Open Access Journals (Sweden)

    ahmad nezami

    2009-06-01

    Full Text Available In order to evaluate the electrolyte leakage as an indicator of freezing injury in colza (Brassica napus L. genotypes under controlled conditions, a trial carried out at the green house of College of Agriculture, Ferdowsi University of Mashhad. In this study 10 rapeseed genotypes, with 5 temperatures (0, -4, -8, -12 and -16 °C on subplot and acclimation and non acclimation on main plot were evaluated on RCD factorial split plot with two replications. Plants were kept until 3-5 leaf stage in green house condition with 23/16 2 °C (day/night and natural photoperiod. Pots were subjected to acclimation (for three weeks or non acclimation that plants immediately frozen.For acclimation treatment after three weeks freezing was done in thermogradient freezer. The cell membrane integrity was measured through electrolyte leakage and the lethal temperature 50 (LT50 of samples also were determined. There were significant differences (p

  2. The co-inheritance of alpha-thalassemia and sickle cell anemia is associated with better hematological indices and lower consultations rate in Cameroonian patients and could improve their survival.

    Directory of Open Access Journals (Sweden)

    Maryam Bibi Rumaney

    Full Text Available BACKGROUND: Co-inheritance of α-thalassemia was reported to be associated with a delayed age of disease onset among Cameroonian Sickle Cell Anemia (SCA patients. The present study aimed to explore the correlation between α-thalassemia, hematological indices, and clinical events in these patients. METHODS AND FINDINGS: We studied 161 Cameroonian SCA patients and 103 controls (59.1% HbAA with median ages of 17.5 and 23 years. RFLP-PCR was used to confirm SCA genotype and to describe haplotypes in the HBB-like genes cluster. Multiplex Gap-PCR was performed to investigate the 3.7 kb α-globin gene deletions. SNaPshot PCR, capillary electrophoresis and cycle sequencing were used for the genotyping of 10 SNPs in BCL11A, HMIP1/2, OR51B5/6 and HBG loci, known to influence HbF levels. Generalised linear regression models adjusted for age, sex and SNPs genotypes was used to investigate effects of α-thalassemia on clinical and hematological indices. The median rate of vaso-occlusive painful crisis and hospitalisations was two and one per year, respectively. Stroke was reported in eight cases (7.4%. Benin haplotype was the most prevalent (66.3%; n = 208 chromosomes. Among patients, 37.3% (n = 60 had at least one 3.7 kb deletion, compared to 10.9% (n = 6 among HbAA controls (p<0.001. Among patients, the median RBC count increased with the number of 3.7 kb deletions [2.6, 3.0 and 3.4 million/dl, with no, one and two deletions (p = 0.01]. The median MCV decreased with the number of 3.7 kb deletion [86, 80, and 68fl, with no, one and two deletions (p<0.0001], as well as median WBC counts [13.2, 10.5 and 9.8×109/L (p<0.0001. The co-inheritance of α-thalassemia was associated with lower consultations rate (p = 0.038. CONCLUSION: The co-inheritance of α-thalassemia and SCA is associated with improved hematological indices, and lower consultations rate in this group of patients. This could possibly improve their survival and explain the

  3. Interaction of Defensins with Model Cell Membranes

    Science.gov (United States)

    Sanders, Lori K.; Schmidt, Nathan W.; Yang, Lihua; Mishra, Abhijit; Gordon, Vernita D.; Selsted, Michael E.; Wong, Gerard C. L.

    2009-03-01

    Antimicrobial peptides (AMPs) comprise a key component of innate immunity for a wide range of multicellular organisms. For many AMPs, activity comes from their ability to selectively disrupt and lyse bacterial cell membranes. There are a number of proposed models for this action, but the detailed molecular mechanism of selective membrane permeation remains unclear. Theta defensins are circularized peptides with a high degree of selectivity. We investigate the interaction of model bacterial and eukaryotic cell membranes with theta defensins RTD-1, BTD-7, and compare them to protegrin PG-1, a prototypical AMP, using synchrotron small angle x-ray scattering (SAXS). The relationship between membrane composition and peptide induced changes in membrane curvature and topology is examined. By comparing the membrane phase behavior induced by these different peptides we will discuss the importance of amino acid composition and placement on membrane rearrangement.

  4. Acute myeloid leukemia (AML)-reactive cytotoxic T lymphocyte clones rapidly expanded from CD8(+) CD62L((high)+) T cells of healthy donors prevent AML engraftment in NOD/SCID IL2Rgamma(null) mice.

    Science.gov (United States)

    Distler, Eva; Wölfel, Catherine; Köhler, Sylvia; Nonn, Marion; Kaus, Nina; Schnürer, Elke; Meyer, Ralf G; Wehler, Thomas C; Huber, Christoph; Wölfel, Thomas; Hartwig, Udo F; Herr, Wolfgang

    2008-04-01

    Current in vitro techniques for isolating leukemia-reactive cytotoxic T lymphocytes (CTLs) from healthy donors are of relatively low efficiency and yield responder populations with unknown biological significance. This study aimed at the development of a more reliable approach, allowing generation and expansion of acute myeloid leukemia (AML)-reactive CTLs using primary in vitro stimulation. We established allogeneic mini-mixed lymphocyte-leukemia cultures (mini-MLLCs) by stimulating donor CD8(+) T cells with human leukocyte antigen (HLA) class I-matched AML blasts in microtiter plates. Before culture, CD8(+) T cells were separated into CD62L((high)+) and CD62L((low)+/neg) subsets enriched for naive/central memory and effector memory cells, respectively. In eight different related and unrelated donor/AML pairs, numerous CTL populations were isolated that specifically lysed myeloid leukemias in association with various HLA-A, -B, or -C alleles. These CTLs expressed T-cell receptors of single Vbeta-chain families, indicating their clonal origin. The majority of CTL clones were obtained from mini-MLLCs initiated with CD62L((high)+) cells. Using antigen-specific stimulation, multiple CTL populations were amplified to 10(8)-10(10) cells within 6 to 8 weeks. Three of four representative CTL clones were capable of completely preventing engraftment of human primary AML blasts in nonobese diabetic/severe combined immune deficient IL2Rgamma(null) mice. The mini-MLLC approach allows the efficient in vitro expansion of AML-reactive CTL clones from CD8(+)CD62L((high)+) precursors of healthy donors. These CTLs can inhibit leukemia engraftment in immunodeficient mice, suggesting their potential biological relevance.

  5. Flow cytometric 96-well microplate-based in vitro micronucleus assay with human TK6 cells: protocol optimization and transferability assessment.

    Science.gov (United States)

    Bryce, Steven M; Avlasevich, Svetlana L; Bemis, Jeffrey C; Tate, Matthew; Walmsley, Richard M; Saad, Frédéric; Van Dijck, Kris; De Boeck, Marlies; Van Goethem, Freddy; Lukamowicz-Rajska, Magdalena; Elhajouji, Azeddine; Dertinger, Stephen D

    2013-04-01

    An automated approach for scoring in vitro micronuclei (MN) has been described in which flow cytometric analysis is combined with compound exposure, processing, and sampling in a single 96-well plate (Bryce SM et al. [2010]: Mutat Res 703:191-199). The current report describes protocol optimization and an interlaboratory assessment of the assay's transferability and reproducibility. In a training phase, the methodology was refined and collaborating laboratories were qualified by repeatedly testing three compounds. Second, a set of 32 chemicals comprised of reference genotoxicants and presumed non-genotoxicants was tested at each of four sites. TK6 cells were exposed to 10 closely spaced compound concentrations for 1.5- to 2-cell population doublings, and were then stained and lysed for flow cytometric analysis. MN frequencies were determined by evaluating ≥ 5,000 cells per replicate well, and several indices of cytotoxicity were acquired. The prevalence of positive results varied according to the MN-fold increase used to signify a genotoxic result, as well as the endpoint used to define a cytotoxicity limit. By varying these parameters, assay sensitivity and specificity values ranged from 82 to 98%, and 86 to 97%, respectively. In a third phase, one laboratory tested a further six genotoxicants and five non-genotoxic apoptosis inducers. In these experiments assay specificity was markedly improved when top concentration selection was based on two cytotoxicity endpoints-relative survival and quantification of ethidium monoazide-positive events. Collectively, the results indicate that the miniaturized assay is transferable across laboratories. The 96-well format consumes considerably less compound than conventional in vitro MN test methods, and the high information content provided by flow cytometry helps guard against irrelevant positive results arising from overt toxicity.

  6. Characterization of the Discoidin Domain Receptor 2 Kinase as a Novel Therapeutic Target for Squamous Cell Lung Cancer

    Science.gov (United States)

    2012-09-01

    Immunoblots were performed using the Nupage System (Invitrogen) per the manufacturer’s protocol. Cells were lysed in 1% Nonidet P - 40 with protease (Roche...based on genetic lesions. J Clin Invest 2009;119:1727– 40 . 33. Du J, Bernasconi P , Clauser KR, Mani DR, Finn SP, Beroukhim R, et al. Bead-based...DDR2 mutations. DDR2 knockdown was associated with a decrease in both cell proliferation and in markers of DDR2 signaling including p -Src and p -STAT3

  7. Hericium erinaceus mushroom extracts protect infected mice against Salmonella Typhimurium-Induced liver damage and mortality by stimulation of innate immune cells.

    Science.gov (United States)

    Kim, Sung Phil; Moon, Eunpyo; Nam, Seok Hyun; Friedman, Mendel

    2012-06-01

    The present study investigated the antibacterial effect of four extracts from the fruitbody of the edible medicinal mushroom Hericium erinaceus (hot water extract, HWE; microwave/50% ethanol extract, MWE; acid extract, ACE; and alkaline extract, AKE) against murine salmonellosis. The extracts had no effect on Salmonella ser. Typhimurium growth in culture. Nor were the extracts toxic to murine macrophage cells, RAW 264.7. HWE and MWE stimulated uptake of the bacteria into the macrophage cells as indicated by increased colony-forming unit (CFU) counts of the contents of the lysed macrophages infected with Salmonella Typhimurium for 30 and 60 min. Two hours postinfection, the bacterial counts increased in the macrophages, but 4 and 8 h postinfection the HWE- and MWE-treated cells showed greater activity against the bacteria than the control. HWE- and MWE-treated noninfected macrophages had altered morphology and elevated inducible nitric oxide (NO) synthase (iNOS) mRNA expression. In the presence of S. Typhimurium, iNOS mRNA expression was further increased, accompanied by an increase in NO production. Histology assays of the livers of mice infected with a sublethal dose (1 × 10(4) CFU) of S. Typhimurium showed that HWE and MWE, administered by daily intraperitoneal injection, protected against necrosis of the liver, a biomarker of in vivo salmonellosis. The lifespans of mice similarly infected with a lethal dose of S. Typhimurium (1 × 10(5) CFU) were significantly extended by HWE and MWE. β-Glucan, known to stimulate the immune system, was previously found to be present in high amounts in the active extracts. These results suggest that the mushroom extract activities against bacterial infection in mice occur through the activation of innate immune cells.

  8. Lyn kinase is activated following thrombopoietin stimulation of the megakaryocytic cell line B1647

    DEFF Research Database (Denmark)

    Santini, Valeria; Scappini, Barbara; Gozzini, Antonella;

    2002-01-01

    BACKGROUND AND OBJECTIVES: B1647 is a cell line derived from bone marrow cells of a patient with acute myeloid leukemia (M2) with a complete erythro-megakaryocytic phenotype and bears both k and p isoforms of c-mpl. Interestingly, spontaneous B1647 cell proliferation is significantly potentiated....... In particular, protein substrates between 50 and 140 kDa appeared to be selectively phosphorylated by TPO. We demonstrated that Jak2, Stat3 and Shc were activated in B1647 cells after TPO, as already shown for different cell lines by other authors. Moreover, Lyn kinase activation was detected. Grb2 co...... by thrombopoietin (TPO). DESIGN AND METHODS: We aimed to evaluate the proliferative signal transduction events following the activation of c-mpl and we stimulated B1647 cells with TPO 40 ng/mL for 3, 7, 15 and 30 minutes; cells were then lysed and whole lysates were immunoprecipitated with anti...

  9. CADM1/TSLC1 Identifies HTLV-1-Infected Cells and Determines Their Susceptibility to CTL-Mediated Lysis.

    Directory of Open Access Journals (Sweden)

    Kiruthika Manivannan

    2016-04-01

    Full Text Available Human T cell lymphotropic virus-1 (HTLV-1 primarily infects CD4+ T cells, causing inflammatory disorders or a T cell malignancy in 5% to 10% of carriers. The cytotoxic T lymphocyte (CTL response is a key factor that controls the viral load and thus the risk of disease. The ability to detect the viral protein Tax in primary cells has made it possible to estimate the rate at which Tax-expressing infected cells are eliminated by CTLs in persistently infected people. However, most HTLV-1-infected cells are Tax-at a given time, and their immunophenotype is poorly defined. Here, we aimed to identify a cell-surface molecule expressed by both Tax+ and Tax-HTLV-1-infected cells and use it to analyse the CTL response in fresh peripheral blood mononuclear cells. Cell adhesion molecule 1 (CADM1/TSLC1 was the best single marker of HTLV-1 infection, identifying HTLV-1-infected cells with greater sensitivity and specificity than CD25, CCR4 or ICAM-1. CADM1+CD4+ T cells carried a median of 65% of proviral copies in peripheral blood. In a cohort of 23 individuals, we quantified the rate of CTL-mediated killing of Tax+ and Tax-CADM1+ cells. We show that CADM1 expression is associated with enhanced susceptibility of infected cells to CTL lysis: despite the immunodominance of Tax in the CTL response, Tax+CADM1- cells were inefficiently lysed by CTLs. Upregulation of the CADM1 ligand CRTAM on CD8+ T cells correlated with efficient lysis of infected cells. Tax-CADM1+ cells were lysed at a very low rate by autologous CTLs, however, were efficiently killed when loaded with exogenous peptide antigen. High expression of CADM1 on most HTLV-1-infected cells in the face of enhanced CTL counterselection implies that CADM1 confers a strong benefit on the virus.

  10. The nucleocytoplasmic microfilament network in protoplasts from cultured soybean cells is a plastic entity that pervades the cytoplasm except the central vacuole.

    Science.gov (United States)

    Villanueva, Marco A; Schindler, Melvin; Wang, John L

    2005-11-01

    The microfilament network of cultured Glycine max cells (SB-1 line), and protoplasts was visualized with rhodamine-phalloidin under conditions that lysed the protoplast and changed the cell shape. The whole cell had the typical microfilament distribution of a "cage" around the nucleus, from which the large subcortical cables and transvacuolar strands radiated towards the cortex until it reached the cortical microfilament network. Upon cell wall removal, the network conserved its compartmentalization. Thus, the redistribution of the shape where the vacuole becomes a central entity, made the cytoplasm displace peripherally, but the network distribution was conserved. When protoplasts were lysed in a low osmotic medium, the vacuoles were gradually released intact. Under these conditions, the F-actin staining remained within the ghost of the cell, but none was detected in either emerging or almost completely released vacuoles. Most of the released F-actin was found in debris from the cell lysate in the form of microfilaments. When the ghosts were constrained in a coverslip with an air bubble, the shape of the ghost changed accordingly, but the microfilament network distribution remained constant. These results provide further evidence that the vacuole of plant cells does not have detectable associated F-actin. In addition, we demonstrate that the actin microfilament network is a moldable entity that can change its shape but keeps its distribution under constant conditions, in these cultured cells.

  11. Should researchers use single indicators, best indicators, or multiple indicators in structural equation models?

    Directory of Open Access Journals (Sweden)

    Hayduk Leslie A

    2012-10-01

    Full Text Available Abstract Background Structural equation modeling developed as a statistical melding of path analysis and factor analysis that obscured a fundamental tension between a factor preference for multiple indicators and path modeling’s openness to fewer indicators. Discussion Multiple indicators hamper theory by unnecessarily restricting the number of modeled latents. Using the few best indicators – possibly even the single best indicator of each latent – encourages development of theoretically sophisticated models. Additional latent variables permit stronger statistical control of potential confounders, and encourage detailed investigation of mediating causal mechanisms. Summary We recommend the use of the few best indicators. One or two indicators are often sufficient, but three indicators may occasionally be helpful. More than three indicators are rarely warranted because additional redundant indicators provide less research benefit than single indicators of additional latent variables. Scales created from multiple indicators can introduce additional problems, and are prone to being less desirable than either single or multiple indicators.

  12. The biology of NK cells and their receptors affects clinical outcomes after hematopoietic cell transplantation (HCT).

    Science.gov (United States)

    Foley, Bree; Felices, Martin; Cichocki, Frank; Cooley, Sarah; Verneris, Michael R; Miller, Jeffrey S

    2014-03-01

    Natural killer (NK) cells were first identified for their capacity to reject bone marrow allografts in lethally irradiated mice without prior sensitization. Subsequently, human NK cells were detected and defined by their non-major histocompatibility complex (MHC)-restricted cytotoxicity toward transformed or virally infected target cells. Karre et al. later proposed 'the missing self hypothesis' to explain the mechanism by which self-tolerant cells could kill targets that had lost self MHC class I. Subsequently, the receptors that recognize MHC class I to mediate tolerance in the host were identified on NK cells. These class I-recognizing receptors contribute to the acquisition of function by a dynamic process known as NK cell education or licensing. In the past, NK cells were assumed to be short lived, but more recently NK cells have been shown to mediate immunologic memory to secondary exposures to cytomegalovirus infection. Because of their ability to lyse tumors with aberrant MHC class I expression and to produce cytokines and chemokines upon activation, NK cells may be primed by many stimuli, including viruses and inflammation, to contribute to a graft-versus-tumor effect. In addition, interactions with other immune cells support the therapeutic potential of NK cells to eradicate tumor and to enhance outcomes after hematopoietic cell transplantation. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  13. Bispecific T cell engager (BiTE®) antibody constructs can mediate bystander tumor cell killing

    Science.gov (United States)

    Ross, Sandra L.; Sherman, Marika; McElroy, Patricia L.; Lofgren, Julie A.; Moody, Gordon; Baeuerle, Patrick A.; Coxon, Angela

    2017-01-01

    For targets that are homogenously expressed, such as CD19 on cells of the B lymphocyte lineage, immunotherapies can be highly effective. Targeting CD19 with blinatumomab, a CD19/CD3 bispecific antibody construct (BiTE®), or with chimeric antigen receptor T cells (CAR-T) has shown great promise for treating certain CD19-positive hematological malignancies. In contrast, solid tumors with heterogeneous expression of the tumor-associated antigen (TAA) may present a challenge for targeted therapies. To prevent escape of TAA-negative cancer cells, immunotherapies with a local bystander effect would be beneficial. As a model to investigate BiTE®-mediated bystander killing in the solid tumor setting, we used epidermal growth factor receptor (EGFR) as a target. We measured lysis of EGFR-negative populations in vitro and in vivo when co-cultured with EGFR-positive cells, human T cells and an EGFR/CD3 BiTE® antibody construct. Bystander EGFR-negative cells were efficiently lysed by BiTE®-activated T cells only when proximal to EGFR-positive cells. Our mechanistic analysis suggests that cytokines released by BiTE®-activated T-cells induced upregulation of ICAM-1 and FAS on EGFR-negative bystander cells, contributing to T cell-induced bystander cell lysis. PMID:28837681

  14. Pre-clinical evaluation of CD38 chimeric antigen receptor engineered T cells for the treatment of multiple myeloma

    DEFF Research Database (Denmark)

    Drent, Esther; Groen, Richard W. J.; Noort, Willy A. Noort

    2016-01-01

    Adoptive transfer of chimeric antigen receptor-transduced T cells is a promising strategy for cancer immunotherapy. The CD38 molecule, with its high expression on multiple myeloma cells, appears a suitable target for antibody therapy. Prompted by this, we used three different CD38 antibody...... sequences to generate second-generation retroviral CD38- chimeric antigen receptor constructs with which we transduced T cells from healthy donors and multiple myeloma patients. We then evaluated the preclinical efficacy and safety of the transduced T cells. Irrespective of the donor and antibody sequence......, CD38-chimeric antigen receptor-transduced T cells proliferated, produced inflammatory cytokines and effectively lysed malignant cell lines and primary malignant cells from patients with acute myeloid leukemia and multi-drug resistant multiple myeloma in a cell-dose, and CD38-dependent manner, despite...

  15. HPLC analysis of Vicia guard cells indicates that products from photosynthetic carbon fixation and starch hydrolysis have an osmotic role during stomatal opening under blue (BL) and red (RL) light

    Energy Technology Data Exchange (ETDEWEB)

    Talbott, L.D.; Zeiger, E. (Univ. of California, Los Angeles (United States))

    1991-05-01

    HPLC was used to quantify neutral sugars and organic acids in guard cells of sonicated Vicia faba epidermal peels irradiated with BL or RL in the presence of 1 mM KCl. Under photosynthetically inactive, low fluence-rates of BL, guard cells initially accumulate malate and citrate. At later times, sucrose and starch breakdown products such as maltose predominate. Guard cells opening under saturating fluence rates of RL show very little organic acid or maltose accumulation, and accumulate mainly sucrose. Changes in metabolite concentrations were correlated with stomatal apertures in both light treatments. These results support previous observations that light quality modulated alternative mechanisms of osmotic accumulation in guard cells, including K{sup +} uptake, photosynthesis and starch hydrolysis. At 5 mM, KCl suppresses RL but not BL-induced opening. These contrasting KCl treatments can be used to investigate osmoregulatory features in guard cells.

  16. Values of Kp Indices, Ap Indices, Cp Indices, C9 Indices, Sunspot Number, and 10.7 cm Flux

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data file consists of Kp indices, Ap indices, Cp indices, C9 indices, sunspot number, and 10.7 cm flux. The most often requested parameter of this file are the...

  17. Nordic eHealth Indicators

    DEFF Research Database (Denmark)

    2013-01-01

    eHealth indicator and benchmarking activities are rapidly increasing nationally and internationally. The work is rarely based on a transparent methodology for indicator definition. This article describes first results of testing an indicator methodology for defining eHealth indicators, which...

  18. FGF Signaling and Dietary Factors in the Prostate

    Science.gov (United States)

    2006-09-01

    preparation of recombinant baculoviruses for expression in Sf9 insect cells, or pcDNA-zeo-3.1(+) vector for expression in mammalian cells. Pull...down with Ni-beads. Sf9 cells (3Х106) infected with indicated recombinant viruses were lysed with 1 % Triton X-100 in PBS. The cell lysates were...Phosphorylation of FRS2 by FGFR1 kinase. Sf9 insect cells (3X106) co-infected with the indicated FRS2 and FGFR1 bearing baculoviruses were lysed

  19. Indicators for Monitoring Soil Biodiversity

    DEFF Research Database (Denmark)

    Bispo, A.; Cluzeau, D.; Creamer, R.

    2009-01-01

    is made for a set of suitable indicators for monitoring the decline in soil biodiversity (Bispo et al. 2007). These indicators were selected both from a literature review and an inventory of national monitoring programmes. Decline in soil biodiversity was defined as the reduction of forms of life living...... indicators are actually measured.   For monitoring application it was considered in ENVASSO that only three key indicators per soil stress were practical. For indicating biodiversity decline it was difficult to arrive at a small set of indicators due to the complexity of soil biota and functions. Therefore...

  20. Esvaziamento cervical no carcinoma epidermoide de laringe: indicação de esvaziamento eletivo contralateral Neck dissection in squamous cell carcinoma of the larynx: indication of elective contralateral neck dissection

    Directory of Open Access Journals (Sweden)

    Ali Amar

    2012-04-01

    Full Text Available O planejamento terapêutico dos tumores da laringe inclui a possibilidade de esvaziamento cervical uni ou bilateral. OBJETIVO: Avaliar a prevalência e localização de metástases linfonodais bilaterais ou contralaterais nos tumores de laringe. CASUÍSTICA E MÉTODO: Estudo longitudinal retrospectivo de 272 prontuários de pacientes com carcinoma epidermoide de laringe tratados entre 1996 e 2004, selecionados 104 submetidos à cirurgia com esvaziamento cervical. Avaliada a incidência de metástases bilaterais ou contralaterais, de acordo com a localização e extensão do tumor primário, considerando os subsítios anatômicos e a linha mediana. RESULTADOS: A prevalência de metástases contralaterais nos tumores lateralizados foi de 3,5% nas lesões glóticas e 26% nas lesões supraglóticas. As metástases contralaterais foram infrequentes nos casos N0. Tanto no esvaziamento eletivo como no terapêutico, os níveis linfonodais IIa e III foram os mais acometidos. CONCLUSÃO: Nas lesões glóticas lateralizadas, mesmo nos tumores transglóticos, não há necessidade de esvaziamento contralateral eletivo. Nas lesões da supraglote sem metástases ipsilaterais, a incidência de metástases ocultas não justifica o esvaziamento eletivo contralateral. A linha mediana não é um indicador fidedigno do risco de metástases contralaterais nos tumores da laringe.Unilateral or bilateral neck dissection must be considered in the treatment of laryngeal cancer AIM: To evaluate the prevalence of contralateral metastases in larynx cancer and distribution of these metastases according to lymph node levels in the neck. METHOD: Retrospective longitudinal study of 272 charts from patients with squamous cell cancer of the larynx treated between 1996 and 2004; and we selected 104 surgical cases submitted to neck dissection. We evaluated the incidence of bilateral or contralateral metastases, according to the location and extension of the primary tumor

  1. Neutralization of (NK-cell-derived) B-cell activating factor by Belimumab restores sensitivity of chronic lymphoid leukemia cells to direct and Rituximab-induced NK lysis.

    Science.gov (United States)

    Wild, J; Schmiedel, B J; Maurer, A; Raab, S; Prokop, L; Stevanović, S; Dörfel, D; Schneider, P; Salih, H R

    2015-08-01

    Natural killer (NK) cells are cytotoxic lymphocytes that substantially contribute to the therapeutic benefit of antitumor antibodies like Rituximab, a crucial component in the treatment of B-cell malignancies. In chronic lymphocytic leukemia (CLL), the ability of NK cells to lyse the malignant cells and to mediate antibody-dependent cellular cytotoxicity upon Fc receptor stimulation is compromised, but the underlying mechanisms are largely unclear. We report here that NK-cells activation-dependently produce the tumor necrosis factor family member 'B-cell activating factor' (BAFF) in soluble form with no detectable surface expression, also in response to Fc receptor triggering by therapeutic CD20-antibodies. BAFF in turn enhanced the metabolic activity of primary CLL cells and impaired direct and Rituximab-induced lysis of CLL cells without affecting NK reactivity per se. The neutralizing BAFF antibody Belimumab, which is approved for treatment of systemic lupus erythematosus, prevented the effects of BAFF on the metabolism of CLL cells and restored their susceptibility to direct and Rituximab-induced NK-cell killing in allogeneic and autologous experimental systems. Our findings unravel the involvement of BAFF in the resistance of CLL cells to NK-cell antitumor immunity and Rituximab treatment and point to a benefit of combinatory approaches employing BAFF-neutralizing drugs in B-cell malignancies.

  2. Edible Acid-Base Indicators.

    Science.gov (United States)

    Mebane, Robert C.; Rybolt, Thomas R.

    1985-01-01

    Reports on the colors observed during titrations of 15 natural indicators obtained from common fruits and vegetables. These edible indicators can be used for a variety of teacher demonstrations or for simple student experiments. (JN)

  3. Indicating anthropogenic effectson urban water system - indicators and extension

    Science.gov (United States)

    Strauch, G.; Ufz-Team

    2003-04-01

    Urban water systems are polluted by diffusive and direct contribution of anthropogenic activities. Besides industrial contaminants like aromatic and chlorinated HC and other persistent organic compounds, the urban aquatic environment is increasingly polluted by low concentrated but high eco-toxic compounds as pharmaceuticals, fragrances, plasticizers which most have disrupt endocrine functions, and trace elements carried in by surface and sub-surface waste water and seeping processes. This contamination could have a longtime impact on the urban ecosystem and on the human health. The interdisciplinary project on risk assessment of water pollution was initiated to explore new methodologies for assessing human activities on the urban water system and processes among urban watersheds. In a first assumption we used a flow model concept with in- and output and surface water transport represented by the city of Halle, Germany, and the river Saale. The river Saale acts as surface water system collecting waste water inputs along the city traverse. We investigated the anthropogenic effect on the urban water system using the indicators hydrological parameters, compound specific pattern of complex organic substances and trace elements, isotopic signatures of water (H, O) and dissolved substances (sulfate, DIC, nitrate), pathogens, and microbiota. A first balance modeling showed that main ions are not very sensitive concerning the direct urban input into the river. Depending on the discharge of the river in high and low flood stages the load of dissolved matter has no specific urban effect. However, the concentration pattern of fragrances (tonalid, galaxolid) and endocrine disrupters (t-nonylphenol) point to a different pollution along the city traverse: downstream of the sewage plant a higher load was observed in comparison to the upstream passage. Furthermore, a degradation ability of fungi and bacteria occurred in the bank sediments could be detected in lab experiments

  4. In vitro cytocidal effect of lytic peptides on several transformed mammalian cell lines.

    Science.gov (United States)

    Jaynes, J M; Julian, G R; Jeffers, G W; White, K L; Enright, F M

    1989-01-01

    Several types of transformed mammalian cells, derived from established cell lines, were found to be lysed in vitro by three novel lytic peptides (SB-37, SB-37*, and Shiva-1). This is in contrast with the behavior of normal cells, where the observed lytic activity of the peptides is greatly reduced. Based on experiments utilizing compounds which disrupt the cytoskeleton (colchicine and cytochalasin-D), it is surmised that alterations in the cytoskeleton of transformed cells increase their sensitivity to the cytolytic activity exerted by the peptides, primarily by causing a loss of osmotic integrity. Thus, a stable and regenerative cytoskeletal system, as that possessed by normal cells, would seem requisite to withstanding the lytic effects of the peptides.

  5. Bispecific Antibodies that Mediate Killing of Cells Infected with Human Immunodeficiency Virus of Any Strain

    Science.gov (United States)

    Berg, Jorg; Lotscher, Erika; Steimer, Kathelyn S.; Capon, Daniel J.; Baenziger, Jurg; Jack, Hans-Martin; Wabl, Matthias

    1991-06-01

    Although AIDS patients lose human immunodeficiency virus (HIV)-specific cytotoxic T cells, their remaining CD8-positive T lymphocytes maintain cytotoxic function. To exploit this fact we have constructed bispecific antibodies that direct cytotoxic T lymphocytes of any specificity to cells that express gp120 of HIV. These bispecific antibodies comprise one heavy/light chain pair from an antibody to CD3, linked to a heavy chain whose variable region has been replaced with sequences from CD4 plus a second light chain. CD3 is part of the antigen receptor on T cells and is responsible for signal transduction. In the presence of these bispecific antibodies, T cells of irrelevant specificity effectively lyse HIV-infected cells in vitro.

  6. Social Indicators and Social Forecasting.

    Science.gov (United States)

    Johnston, Denis F.

    The paper identifies major types of social indicators and explains how they can be used in social forecasting. Social indicators are defined as statistical measures relating to major areas of social concern and/or individual well being. Examples of social indicators are projections, forecasts, outlook statements, time-series statistics, and…

  7. The SJR indicator: A new indicator of journals' scientific prestige

    CERN Document Server

    Gonzalez-Pereira, Borja; Moya-Anegon, Felix

    2009-01-01

    This paper proposes an indicator of journals' scientific prestige, the SJR indicator, for ranking scholarly journals based on citation weighting schemes and eigenvector centrality to be used in complex and heterogeneous citation networks such Scopus. Its computation methodology is described and the results after implementing the indicator over Scopus 2007 dataset are compared to an ad-hoc Journal Impact Factor both generally and inside specific scientific areas. The results showed that SJR indicator and JIF distributions fitted well to a power law distribution and that both metrics were strongly correlated, although there were also major changes in rank. There was an observable general trend that might indicate that SJR indicator values decreased certain JIF values whose citedeness was greater than would correspond to their scientific influence.

  8. Herbal indicators as a substituent to synthetic indicators

    OpenAIRE

    Bhagat Vishwas; Patil Radheshyam; Channekar P; Shetty S; Akarte Atul

    2008-01-01

    In acid - base titrations, indicators are used to show sharp color change at intervals of pH. Natural pigments in plants are highly colored substances and may show color changes with variation of pH. An attempt has been made to investigate the indicator activity of methanolic extract of flower pigments and to replace synthetic indicators as they have certain disadvantages like chemical pollution, availability problems and high cost. Methanolic extract of Rosa indica (L) from family Rosaceae a...

  9. Cancer rehabilitation indicators for Europe

    DEFF Research Database (Denmark)

    Baili, Paolo; Hoekstra-Weebers, Josette; Van Hoof, Elke

    2013-01-01

    Little is known of cancer rehabilitation needs in Europe. EUROCHIP-3 organised a group of experts to propose a list of population-based indicators used for describing cancer rehabilitation across Europe. The aim of this study is to present and discuss these indicators. A EUROCHIP-3 expert panel...... reached agreement on two types of indicators. (a) Cancer prevalence indicators. These were proposed as a means of characterising the burden of cancer rehabilitation needs by time from diagnosis and patient health status. These indicators can be estimated from cancer registry data or by collecting data...... on follow-up and treatments for samples of cases archived in cancer registries. (b) Indicators of rehabilitation success. These include: return to work, quality of life, and satisfaction of specific rehabilitation needs. Studies can be performed to estimate these indicators in individual countries...

  10. Biological monitoring of radiation using indicator plants

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jin Kyoo; Chun, Ki Jung; Kim, Kook Chan; Kim, In Kyoo; Song, Heui Sub [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of)

    1994-12-01

    Some clones of Tradescantia had dose response relationship involving somatic mutations such as appearance of pink, colorless or giant cell, and/or loss of reproductive integrity of stamen hair cells when exposed to radiation. Since Tradescantia could respond to radiation level as low as human being could be exposed to, it could play an important role as scientific tool of botanical tester for radiation. Especially TSH system can be easily applied to in situ monitoring of radiation by virtue of its excellent radiation indicator ship and simpleness in detection of mutations by radiation. 10 figs, 6 tabs, 19 refs. (Author).

  11. New insights into estrogenic regulation of O(6)-methylguanine DNA-methyltransferase (MGMT) in human breast cancer cells: Co-degradation of ER-α and MGMT proteins by fulvestrant or O(6)-benzylguanine indicates fresh avenues for therapy.

    Science.gov (United States)

    Paranjpe, Ameya; Bailey, Nathan I; Konduri, Santhi; Bobustuc, George C; Ali-Osman, Francis; Yusuf, Mohd A; Punganuru, Surendra R; Madala, Hanumantha Rao; Basak, Debasish; Mostofa, Agm; Srivenugopal, Kalkunte S

    2016-09-01

    Endocrine therapy using estrogen receptor-α (ER-α) antagonists for attenuating horm2one-driven cell proliferation is a major treatment modality for breast cancers. To exploit any DNA repair deficiencies associated with endocrine therapy, we investigated the functional and physical interactions of ER-α with O(6)-methylguanine DNA methyltransferase (MGMT), a unique DNA repair protein that confers tumor resistance to various anticancer alkylating agents. The ER-α -positive breast cancer cell lines (MCF-7, T47D) and ER- negative cell lines (MDAMB-468, MDAMB-231), and established inhibitors of ER-α and MGMT, namely, ICI-182,780 (Faslodex) and O(6)-benzylguanine, respectively, were used to study MGMT- ER interactions. The MGMT gene promoter was found to harbor one full and two half estrogen-responsive elements (EREs) and two antioxidant-responsive elements (AREs). MGMT expression was upregulated by estrogen, downregulated by tamoxifen in Western blot and promoter-linked reporter assays. Similarly, both transient and stable transfections of Nrf-2 (nuclear factor-erythroid 2-related factor-2) increased the levels of MGMT protein and activity 3 to 4-fold reflecting novel regulatory nodes for this drug-resistance determinant. Of the different ER-α antagonists tested, the pure anti-estrogen fulvestrant was most potent in inhibiting the MGMT activity in a dose, time and ER-α dependent manner, similar to O(6)-benzylguanine. Interestingly, fulvestrant exposure led to a degradation of both ER-α and MGMT proteins and O(6)-benzylguanine also induced a specific loss of ER-α and MGMT proteins in MCF-7 and T47D breast cancer cells with similar kinetics. Immunoprecipitation revealed a specific association of ER-α and MGMT proteins in breast cancer cells. Furthermore, silencing of MGMT gene expression triggered a decrease in the levels of both MGMT and ER-α proteins. The involvement of proteasome in the drug-induced degradation of both proteins was also demonstrated

  12. New insights into estrogenic regulation of O6-methylguanine DNA-methyltransferase (MGMT) in human breast cancer cells: Co-degradation of ER-α and MGMT proteins by fulvestrant or O6-benzylguanine indicates fresh avenues for therapy

    Science.gov (United States)

    Paranjpe, Ameya; Bailey, Nathan I.; Konduri, Santhi; Bobustuc, George C.; Ali-Osman, Francis; Yusuf, Mohd. A.; Punganuru, Surendra R.; Madala, Hanumantha Rao; Basak, Debasish; Mostofa, AGM; Srivenugopal, Kalkunte S.

    2016-01-01

    Abstract Endocrine therapy using estrogen receptor-α (ER-α) antagonists for attenuating horm2one-driven cell proliferation is a major treatment modality for breast cancers. To exploit any DNA repair deficiencies associated with endocrine therapy, we investigated the functional and physical interactions of ER-α with O6-methylguanine DNA methyltransferase (MGMT), a unique DNA repair protein that confers tumor resistance to various anticancer alkylating agents. The ER-α -positive breast cancer cell lines (MCF-7, T47D) and ER- negative cell lines (MDAMB-468, MDAMB-231), and established inhibitors of ER-α and MGMT, namely, ICI-182,780 (Faslodex) and O6-benzylguanine, respectively, were used to study MGMT- ER interactions. The MGMT gene promoter was found to harbor one full and two half estrogen-responsive elements (EREs) and two antioxidant-responsive elements (AREs). MGMT expression was upregulated by estrogen, downregulated by tamoxifen in Western blot and promoter-linked reporter assays. Similarly, both transient and stable transfections of Nrf-2 (nuclear factor-erythroid 2-related factor-2) increased the levels of MGMT protein and activity 3 to 4-fold reflecting novel regulatory nodes for this drug-resistance determinant. Of the different ER-α antagonists tested, the pure anti-estrogen fulvestrant was most potent in inhibiting the MGMT activity in a dose, time and ER-α dependent manner, similar to O6-benzylguanine. Interestingly, fulvestrant exposure led to a degradation of both ER-α and MGMT proteins and O6-benzylguanine also induced a specific loss of ER-α and MGMT proteins in MCF-7 and T47D breast cancer cells with similar kinetics. Immunoprecipitation revealed a specific association of ER-α and MGMT proteins in breast cancer cells. Furthermore, silencing of MGMT gene expression triggered a decrease in the levels of both MGMT and ER-α proteins. The involvement of proteasome in the drug-induced degradation of both proteins was also demonstrated

  13. Mycobacterium tuberculosis CwsA overproduction modulates cell division and cell wall synthesis.

    Science.gov (United States)

    Plocinski, P; Martinez, L; Sarva, K; Plocinska, R; Madiraju, M; Rajagopalan, M

    2013-12-01

    We recently showed that two small membrane proteins of Mycobacterium tuberculosis, CwsA and CrgA, interact with each other, and that loss of CwsA in M. smegmatis is associated with defects in the cell division and cell wall synthesis processes. Here we show that CwsA overproduction also affected growth, cell division and cell shape of M. smegmatis and M. tuberculosis. CwsA overproduction in M. tuberculosis led to increased sensitivity to cefsulodin, a penicillin-binding protein (PBP) 1A/1B targeting beta (β) -lactam, but was unaffected by other β-lactams and vancomycin. A M. smegmatis cwsA overexpressing strain showed bulgy cells, increased fluorescent vancomycin staining and altered localization of Wag31-mCherry fusion protein. However, the levels of phosphorylated Wag31, important for optimal peptidoglycan synthesis and growth in mycobacteria, were not affected. Interestingly, CwsA overproduction in E. coli led to the formation of large rounded cells that eventually lysed whereas the overproduction of FtsZ along with CwsA reversed this phenotype. Together, our results emphasize that optimal levels of CwsA are required for regulated cell wall synthesis, hence maintenance of cell shape, and that CwsA likely interacts with and modulates the activities of other cell wall synthetic components including PBPs.

  14. Carbon nanotubes for voltage reduction and throughput enhancement of electrical cell lysis on a lab-on-a-chip

    Energy Technology Data Exchange (ETDEWEB)

    Shahini, Mehdi; Yeow, John T W, E-mail: jyeow@uwaterloo.ca [University of Waterloo, 200 University Avenue West, Waterloo, ON (Canada)

    2011-08-12

    We report on the enhancement of electrical cell lysis using carbon nanotubes (CNTs). Electrical cell lysis systems are widely utilized in microchips as they are well suited to integration into lab-on-a-chip devices. However, cell lysis based on electrical mechanisms has high voltage requirements. Here, we demonstrate that by incorporating CNTs into microfluidic electrolysis systems, the required voltage for lysis is reduced by half and the lysis throughput at low voltages is improved by ten times, compared to non-CNT microchips. In our experiment, E. coli cells are lysed while passing through an electric field in a microchannel. Based on the lightning rod effect, the electric field strengthened at the tip of the CNTs enhances cell lysis at lower voltage and higher throughput. This approach enables easy integration of cell lysis with other on-chip high-throughput sample-preparation processes.

  15. Uncertainty in magnetic activity indices

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Magnetic activity indices are widely used in theoretical studies of solar-terrestrial coupling and space weather prediction. However, the indices suffer from various uncertainties, which limit their application and even mislead to incorrect conclu-sion. In this paper we analyze three most popular indices, Kp, AE and Dst. Three categories of uncertainties in magnetic indices are discussed: "data uncertainty" originating from inadequate data processing, "station uncertainty" caused by in-complete station covering, and "physical uncertainty" stemming from unclear physical mechanism. A comparison between magnetic disturbances and related indices indicate that the residual Sq will cause an uncertainty of 1―2 in K meas-urement, the uncertainty in saturated AE is as much as 50%, and the uncertainty in Dst index caused by the partial ring currents is about a half of the partial ring cur-rent.

  16. Uncertainty in magnetic activity indices

    Institute of Scientific and Technical Information of China (English)

    XU WenYao

    2008-01-01

    Magnetic activity indices are widely used in theoretical studies of solar-terrestrial coupling and space weather prediction. However, the indices suffer from various uncertainties, which limit their application and even mislead to incorrect conclu-sion. In this paper we analyze three most popular indices, Kp, AE and Dst. Three categories of uncertainties in magnetic indices are discussed: "data uncertainty" originating from inadequate data processing, "station uncertainty" caused by in-complete station covering, and "physical uncertainty" stemming from unclear physical mechanism. A comparison between magnetic disturbances and related indices indicate that the residual Sq will cause an uncertainty of 1-2 in K meas-urement, the uncertainty in saturated AE is as much as 50%, and the uncertainty in Dst index caused by the partial ring currents is about a half of the partial ring cur-rent.

  17. Herbal indicators as a substituent to synthetic indicators

    Directory of Open Access Journals (Sweden)

    Bhagat Vishwas

    2008-01-01

    Full Text Available In acid - base titrations, indicators are used to show sharp color change at intervals of pH. Natural pigments in plants are highly colored substances and may show color changes with variation of pH. An attempt has been made to investigate the indicator activity of methanolic extract of flower pigments and to replace synthetic indicators as they have certain disadvantages like chemical pollution, availability problems and high cost. Methanolic extract of Rosa indica (L from family Rosaceae and Hibiscus rosa-sinensis (L from family Malvaceae gives sharp and intense color change as compared to phenolphthalein and methyl orange. Herbal indicators are evaluated by using strong acid - strong base, strong acid - weak base, weak acid - strong base, and weak acid - weak base. In all these titrations the extract was found to be very useful and accurate for indicating the equivalence point (neutralization point. From photochemical investigation and qualitative chemical tests for Rosa indica and Hibiscus rosa-sinensis shows presence of anthocyanins; which are the pigmented flavonoids; which may be the reason for its activity as an indicator. Apart from the above advantages the isolation of pure compounds possessing indicator′s properties help to know the mechanism by which they shows indicator′s properties and new theories of indicators could be established.

  18. Cell-to-cell spread and massive vacuole formation after Cryptococcus neoformans infection of murine macrophages

    Directory of Open Access Journals (Sweden)

    Casadevall Arturo

    2007-08-01

    Full Text Available Abstract Background The interaction between macrophages and Cryptococcus neoformans (Cn is critical for containing dissemination of this pathogenic yeast. However, Cn can either lyse macrophages or escape from within them through a process known as phagosomal extrusion. Both events result in live extracellular yeasts capable of reproducing and disseminating in the extracellular milieu. Another method of exiting the intracellular confines of cells is through host cell-to-cell transfer of the pathogen, and this commonly occurs with the human immuno-deficiency virus (HIV and CD4+ T cells and macrophages. In this report we have used time-lapse imaging to determine if this occurs with Cn. Results Live imaging of Cryptococcus neoformans interactions with murine macrophages revealed cell-to-cell spread of yeast cells from infected donor cells to uninfected cells. Although this phenomenon was relatively rare its occurrence documents a new capacity for this pathogen to infect adjacent cells without exiting the intracellular space. Cell-to-cell spread appeared to be an actin-dependent process. In addition, we noted that cryptococcal phagosomal extrusion was followed by the formation of massive vacuoles suggesting that intracellular residence is accompanied by long lasting damage to host cells. Conclusion C. neoformans can escape the intracellular confines of macrophages in an actin dependent manner by cell-to-cell transfer of the yeast leading to infection of adjacent cells. In addition, complete extrusion of internalized Cn cells can lead to the formation of a massive vacuole which may be a sign of damage to the host macrophage. These observations document new outcomes for the interaction of C. neoformans with host cells that provide precedents for cell biological effects that may contribute to the pathogenesis of cryptococcal infections.

  19. Chronic Inflammation and  T Cells

    Directory of Open Access Journals (Sweden)

    Nathan S Fay

    2016-05-01

    Full Text Available The epithelial tissues of the skin, lungs, reproductive tract, and intestines are the largest physical barriers the body has to protect against infection. Epithelial tissues are woven with a matrix of immune cells programmed to mobilize the host innate and adaptive immune responses. Included among these immune cells are  T cells that are unique in their TCR usage, location, and functions in the body. Stress reception by  T cells as a result of traumatic epithelial injury, malignancy, and/or infection induces  T cell activation. Once activated,  T cells function to repair tissue, induce inflammation, recruit leukocytes, and lyse cells. Many of these functions are mediated via the production of cytokines and growth factors upon  T cell activation. Pathogenesis of many chronic inflammatory diseases involve  T cells; some of which are exacerbated by their presence, while others are improved.  T cells require a delicate balance between their need for acute inflammatory mediators to function normally and the detrimental impact imparted by chronic inflammation. This review will focus on the recent progress made in understanding how epithelial  T cells influence the pathogenesis of chronic inflammatory diseases and how a balance between acute and chronic inflammation impacts  T cell function. Future studies will be important to understand how this balance is achieved.

  20. Wacky Patents Meet Economic Indicators

    DEFF Research Database (Denmark)

    Czarnitzki, Dirk; Hussinger, Katrin; Schneider, Cédric

    2011-01-01

    We investigate whether standard indicators can distinguish between “wacky” patents and a control group. Forward citations are good predictors of importance. However, “wacky” patents have higher originality, generality and citation lags, suggesting that these indicators should be interpreted...

  1. Lexical-Grammatical Pragmatic Indicators.

    Science.gov (United States)

    Blum-Kulka, Shoshana; Levenston, Edward A.

    1987-01-01

    A study of the interlanguage pragmatics of learners of Hebrew and English (L2s) focuses on pragmatic indicators used in requests and apologies (situations in Appendix). Deviations from native-speaker norms in the speech of non-native speakers are discussed. Results suggest L2s' misuse of pragmatic indicators can have serious interactional…

  2. Rapid Benefit Indicators (RBI) webinar

    Science.gov (United States)

    RBI process for assessing restoration sites using non-monetary benefit indicators. The RBI approach uses readily-available data to estimate and quantify benefits to people around an ecological restoration site using indicators of nature’s value to people.

  3. Decisiveness indices are semiindices: addendum

    OpenAIRE

    Freixas Bosch, Josep; Pons Vallès, Montserrat

    2017-01-01

    In the paper Decisiveness indices are semiindices (Freixas and Pons, 2016) it was shown that any decisiveness index obtained from an anonymous probability distribution is a semiindex, and that the converse is not true. In this note we characterize the semiindices which are indices of decisiveness. Peer Reviewed

  4. Quality Indicators for Learning Analytics

    Science.gov (United States)

    Scheffel, Maren; Drachsler, Hendrik; Stoyanov, Slavi; Specht, Marcus

    2014-01-01

    This article proposes a framework of quality indicators for learning analytics that aims to standardise the evaluation of learning analytics tools and to provide a mean to capture evidence for the impact of learning analytics on educational practices in a standardised manner. The criteria of the framework and its quality indicators are based on…

  5. Geomagnetic Kn, Ks, Km Indices

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — A full description of the indices Kn, Ks, Km is given in a monography,"Indices Kn, Ks et Km, 1964-1967", edited in 1968 by the Centre National de la Recherche...

  6. Vegetation indices as indicators of damage by the sunn pest ...

    African Journals Online (AJOL)

    SERVER

    2008-01-18

    Jan 18, 2008 ... structure insensitive pigment index (SIPI) were chosen out of 19 indices initially tested. The NDVI ... Because chlorophyll content plays a direct role in photosynthesis ... near infrared (NIR) reflectance from its leaves. Jensen.

  7. Increased IgG4 responses to multiple food and animal antigens indicate a polyclonal expansion and differentiation of pre-existing B cells in IgG4-related disease

    OpenAIRE

    Culver, EL; Vermeulen, E.; Makuch, M; Leeuwen, A. van; Sadler, R.; Cargill, T; Klenerman, P; Aalberse, RC; van Ham, SM; Barnes, E; Rispens, T

    2015-01-01

    Background: IgG4-related disease (IgG4-RD) is a systemic fibroinflammatory condition, characterised by an elevated serum IgG4 concentration and abundant IgG4-positive plasma cells in the involved organs. An important question is whether the elevated IgG4 response is causal or a reflection of immune-regulatory mechanisms of the disease. Objectives: To investigate if the IgG4 response in IgG4-RD represents a generalised polyclonal amplification by examining the response to common environmental ...

  8. Power Quality Indices Estimation Platform

    Directory of Open Access Journals (Sweden)

    Eliana I. Arango-Zuluaga

    2013-11-01

    Full Text Available An interactive platform for estimating the quality indices in single phase electric power systems is presented. It meets the IEEE 1459-2010 standard recommendations. The platform was developed in order to support teaching and research activities in electric power quality. The platform estimates the power quality indices from voltage and current signals using three different algorithms based on fast Fourier transform (FFT, wavelet packet transform (WPT and least squares method. The results show that the algorithms implemented are efficient for estimating the quality indices of the power and the platform can be used according to the objectives established. 

  9. The politics of SEA indicators

    DEFF Research Database (Denmark)

    Gao, Jingjing; Kørnøv, Lone; Christensen, Per

    2013-01-01

    The use of indicators is not only technical and science-led, but also a value-laden social process, and thus concerns public participation, political judgment and decision-making. This article approaches the Chinese SEA indicator system from a science-policy interface and aims at: 1) contributing...... to the general recognition of indicators functioning at science-policy interfaces in SEA, and 2) analysing, through a Chinese case-study, to what extent national guidelines mediate the science-policy interaction. The overall finding is a strong emphasis on technical/science aspects found in the Chinese SEA...

  10. Hearing Aid with Visual Indicator

    DEFF Research Database (Denmark)

    2004-01-01

    is provided for powering the signal receiving part, the signal path and the audio transducer, and where further means are provided for assessing the function of the hearing aid and for generating an electrical indication signal which indicates the function of the hearing aid and where further means...... are provided for intermittently generating a power signal in response to the electrical indication signal and where means are provided for converting the power signal into a light signal, such that the light signal is visible from outside the hearing aid....

  11. Sustainable Transport and Performance Indicators

    DEFF Research Database (Denmark)

    Gudmundsson, Henrik

    2004-01-01

    Sustainable transport has become a major policy concern in recent years. The term suggests a need to consider transport in a comprehensive perspective including the well-being of both present and future generations, and taking environmental, social and economic issues into account. Indicators...... in forecasting and tracking the performance of transport systems increasingly guide sustainable transport policy making. This review explores the implications of the sustainable transport” concept and how it can be made operational with indicators. A number of indicator systems are reviewed, and some strengths...... and weaknesses are highlighted with regard to the support they provide for sustainable transport development....

  12. Indicators of Difficulty in Translation

    DEFF Research Database (Denmark)

    Dragsted, Barbara

    2012-01-01

    This article sets out to investigate the correlation between indicators of difficulty observable in translation product and translation process data respectively. It has been claimed that the number of alternative renditions in the target text across a group of subjects translating the same source...... text item indicates the degree of cognitive effort. We identified words with high versus low target text variability across eight subjects and related these to various indicators of difficulty observable in process data from eye-tracking and keystroke logging: number of fixations, gaze time, pauses...

  13. Human dendritic cells derived from embryonic stem cells stably modified with CD1d efficiently stimulate antitumor invariant natural killer T cell response.

    Science.gov (United States)

    Zeng, Jieming; Wang, Shu

    2014-01-01

    Invariant natural killer T (iNKT) cells are a unique lymphocyte subpopulation that mediates antitumor activities upon activation. A current strategy to harness iNKT cells for cancer treatment is endogenous iNKT cell activation using patient-derived dendritic cells (DCs). However, the limited number and functional defects of patient DCs are still the major challenges for this therapeutic approach. In this study, we investigated whether human embryonic stem cells (hESCs) with an ectopically expressed CD1d gene could be exploited to address this issue. Using a lentivector carrying an optimized expression cassette, we generated stably modified hESC lines that consistently overexpressed CD1d. These modified hESC lines were able to differentiate into DCs as efficiently as the parental line. Most importantly, more than 50% of such derived DCs were CD1d+. These CD1d-overexpressing DCs were more efficient in inducing iNKT cell response than those without modification, and their ability was comparable to that of DCs generated from monocytes of healthy donors. The iNKT cells expanded by the CD1d-overexpressing DCs were functional, as demonstrated by their ability to lyse iNKT cell-sensitive glioma cells. Therefore, hESCs stably modified with the CD1d gene may serve as a convenient, unlimited, and competent DC source for iNKT cell-based cancer immunotherapy.

  14. Astrocyte cytolysis by MHC class II-specific mouse T cell clones.

    Science.gov (United States)

    Reder, A T; Lascola, C D; Flanders, S A; Maimone, D; Jensen, M A; Skias, D D; Lancki, D W

    1993-08-01

    The brain is "immunologically privileged," in part because class I and II MHC antigens are not normally present on glia or neurons. However, under certain conditions such as transplantation, glial cells express MHC proteins at levels sufficient for glia to become targets of immune responses. Cultured astrocytes expressing very low levels of MHC class I protein are killed efficiently by MHC class I antigen-specific CTL. Mouse brain allografts, however, are rejected by CD4+ T cells that are likely to be class II MHC-specific. The level of expression of MHC class II antigen needed to trigger specific killing of astrocytes by CD4+ T cells, independent of exogenous antigen, has not been studied. We examined the role of glial class II MHC in the lysis of cultured neonatal mouse astrocytes by an alloreactive murine CD4+ CTL alone. IFN-gamma induced functionally relevant increases in MHC class II antigen on target cells. Astrocytes were lysed by the CD4+ clone only when class II MHC antigens reached levels readily detectable by flow cytometry. MHC class II expression and lysis increased when astrocytes were coincubated with IFN-gamma and TNF-alpha. Conversely, lysis decreased when class II expression was downregulated by IFN-alpha/beta or dbcAMP. Cytolysis by CD4+ clones was blocked by antibodies to CD4 and LFA-1 on T cells, and to ICAM-1 and class II molecules on astrocytes. The role of LFA-1 in CD4+ cell-mediated lysis was greater than that of LFA-1/ICAM-1 in CD8+ T cell-mediated lysis. CD4+ cells may lyse activated astrocytes when the immune privilege of the brain is compromised as in transplantation, tumors, and inflammatory diseases.

  15. Immunologic targeting of FOXP3 in inflammatory breast cancer cells.

    Directory of Open Access Journals (Sweden)

    Smita Nair

    Full Text Available The forkhead transcription factor FOXP3 is necessary for induction of regulatory T lymphocytes (Tregs and their immunosuppressive function. We have previously demonstrated that targeting Tregs by vaccination of mice with murine FOXP3 mRNA-transfected dendritic cells (DCs elicits FOXP3-specific T cell responses and enhances tumor immunity. It is clear that FOXP3 expression is not restricted to T-cell lineage and herein, using RT-PCR, flow cytometry, and western immunoblot we demonstrate for the first time that FOXP3 is expressed in inflammatory breast cancer (IBC cells, SUM149 (triple negative, ErbB1-activated and SUM190 (ErbB2-overexpressing. Importantly, FOXP3-specific T cells generated in vitro using human FOXP3 RNA-transfected DCs as stimulators efficiently lyse SUM149 cells. Interestingly, an isogenic model (rSUM149 derived from SUM149 with an enhanced anti-apoptotic phenotype was resistant to FOXP3-specific T cell mediated lysis. The MHC class I cellular processing mechanism was intact in both cell lines at the protein and transcription levels suggesting that the resistance to cytolysis by rSUM149 cells was not related to MHC class I expression or to the MHC class I antigen processing machinery in these cells. Our data suggest that FOXP3 may be an effective tumor target in IBC cells however increased anti-apoptotic signaling can lead to immune evasion.

  16. EJSCREEN Environmental Indicators 2015 Internal

    Data.gov (United States)

    U.S. Environmental Protection Agency — There are 12 EJ Indexes in EJSCREEN reflecting the 12 environmental indicators. The EJ Index names are: Particulate Matter (PM2.5), Ozone, NATA Diesel PM, NATA...

  17. EJSCREEN Environmental Indicators 2016 Public

    Data.gov (United States)

    U.S. Environmental Protection Agency — There are eleven EJ Indexes in EJSCREEN reflecting the 11 environmental indicators. The EJ Index names are: Particulate Matter (PM2.5), Ozone, NATA Diesel PM, NATA...

  18. Investigation of Some Metallochromic Indicators.

    Science.gov (United States)

    Jones, A. V.; Nelson, M.

    1979-01-01

    Presents some introductory experiments involving the reaction between EDTA and several metal ions using metallochromic indicators. The experiments represent familiar reactions and are followed by suggestions for extension into areas of discovery work by students. (Author/SA)

  19. Sewage-pollution indicator bacteria

    Digital Repository Service at National Institute of Oceanography (India)

    Ramaiah, N.; Rodrigues, V.; Alwares, E.; Rodrigues, C.; Baksh, R.; Jayan, S.; Mohandass, C.

    Spatial distribution and annual cycle of sewage pollution indicator (total coliforms and total fecal coliforms) and human pathogenic bacteria (Escherichia coli and Streptococcus faecalis) in water and sediment samples in the Mandovi and Zuari...

  20. EJSCREEN Environmental Indicators 2015 Public

    Data.gov (United States)

    U.S. Environmental Protection Agency — There are eight EJ Indexes in EJSCREEN reflecting the 8 environmental indicators. The eight EJ Index names are: Particulate Matter (PM2.5), Ozone Traffic Proximity...

  1. Fuzzy indicators for customer retention

    National Research Council Canada - National Science Library

    Valenzuela-Fernández, Leslier; Nicolas, Carolina; Gil-Lafuente, Jaime; Merigó, José M

    2016-01-01

    .... Nevertheless, one cannot ignore the existence of a gap on how to measure this relationship. Following this idea, this study proposes six fuzzy key performance indicators that aims to measure customer retention and loyalty of the portfolio...

  2. Atlantic Coast Environmental Indicators Consortium

    Data.gov (United States)

    Federal Laboratory Consortium — n 2000, the US EPA granted authority to establish up to five Estuarine Indicator Research Programs. These Programs were designed to identify, evaluate, recommend and...

  3. EJSCREEN Demographic Indicators 2016 Public

    Data.gov (United States)

    U.S. Environmental Protection Agency — EJSCREEN uses demographic factors as very general indicators of a community's potential susceptibility to the types of environmental factors included in this...

  4. EJSCREEN Demographic Indicators 2015 Internal

    Data.gov (United States)

    U.S. Environmental Protection Agency — EJSCREEN uses demographic factors as very general indicators of a community's potential susceptibility to the types of environmental factors included in this...

  5. EJSCREEN Demographic Indicators 2015 Public

    Data.gov (United States)

    U.S. Environmental Protection Agency — EJSCREEN uses demographic factors as very general indicators of a community's potential susceptibility to the types of environmental factors included in this...

  6. Solar Indices - Solar Radio Flux

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Collection includes a variety of indices related to solar activity contributed by a number of national and private solar observatories located worldwide. This...

  7. Performance Indicators of Operating Reactors

    Data.gov (United States)

    Nuclear Regulatory Commission — A list of Performance Indicators (PI) that are reported to the NRC by licensees at the end of each quarter in accordance with Inspection Manual Chapters (IMC) 0608,...

  8. Cognitive Value of Financial Indicators

    Directory of Open Access Journals (Sweden)

    Gabriela Cristina FLOREA

    2012-11-01

    Full Text Available Economic and Financial indicators are very important tools in the assessment of the economic entity's well-being. They Provide permanent monitoring of the significance of the issues related to the economic activity's deployment. The main Objectives tracked by the process of managing the company with the help of Economic and Financial indicators is the activity's continuity, ensuring liquidity and Achieving positive results.

  9. Current Indications for Coronary Arteriography

    OpenAIRE

    Dodek, Arthur

    1981-01-01

    Coronary arteriography is not necessary in all patients with angina pectoris. Angiography is indicated to define coronary anatomy in patients who have persistent angina despite full medication. It is also indicated in patients with a markedly positive stress test, unstable angina pectoris, positive stress test following myocardial infarction, variants of angina, ventricular aneurysm complications, and in the young coronary patient. Coronary arteriography may clarify the diagnosis in patients ...

  10. Indicators of Information Society Measurement :

    Directory of Open Access Journals (Sweden)

    Hind Elwy

    2006-09-01

    Full Text Available The indicator of information society describe the infrastructure of information and communication technology ; as well as it’s use and it’s production in different estate of society. The importance economic and social of tic is crescent in modern society. and the presentation of tendency inform above the situation of information society . in this article we want to describe the indicator of tic in Algeria according to librarian’s vision in Mentouri university

  11. Sustainable development indicators for cities

    Directory of Open Access Journals (Sweden)

    Sergey Nikolayevich Bobylev

    2014-09-01

    Full Text Available The assessment of urban population’s life quality implies an investigation of all factors defining it: economic, social and ecological. The development of the corresponding indicators of sustainable urban development is necessary. The majority of the cities in the world and this country show unsustainable development at present time. In the article, the world and Russian experience of development of indicators of sustainable urban development is considered. In the article, opportunities of adaptation of approaches to these indicators’ development on the basis of Human Development Index developed by United Nations Development Program and an index of Adjusted Net Savings of the World Bank for Russia are considered. The authors propose a new integrated index of sustainability for Russian cities. It is based on the concept and methodology of the Adjusted Net Savings index. In order to evaluate the sustainability of urban development taking into account economic, social, and ecological factors, the authors propose applying three corresponding sub-indexes: gross capital, expenses on human capital development, and damage from environmental pollution in the cities. In the article, the authors’ set of indicators for Russian cities is proposed. It reflects the most acute problems of sustainable urban development in Russia and the quality of life in cities; also it corresponds to Russian statistics. 21 key indicators reflecting important economic, social, and ecological urban priorities are proposed. Indicators are divided into nine groups: economic indicators; energy efficiency; transport; social and institutional indicators; air and climate; water resources; waste; especially protected natural territories; noise influence. Proposed indicators for cities allow more adequately assess trends of urbanized space shaping and quality of life

  12. Energy Efficiency Indicators Methodology Booklet

    Energy Technology Data Exchange (ETDEWEB)

    Sathaye, Jayant; Price, Lynn; McNeil, Michael; de la rue du Can, Stephane

    2010-05-01

    This Methodology Booklet provides a comprehensive review and methodology guiding principles for constructing energy efficiency indicators, with illustrative examples of application to individual countries. It reviews work done by international agencies and national government in constructing meaningful energy efficiency indicators that help policy makers to assess changes in energy efficiency over time. Building on past OECD experience and best practices, and the knowledge of these countries' institutions, relevant sources of information to construct an energy indicator database are identified. A framework based on levels of hierarchy of indicators -- spanning from aggregate, macro level to disaggregated end-use level metrics -- is presented to help shape the understanding of assessing energy efficiency. In each sector of activity: industry, commercial, residential, agriculture and transport, indicators are presented and recommendations to distinguish the different factors affecting energy use are highlighted. The methodology booklet addresses specifically issues that are relevant to developing indicators where activity is a major factor driving energy demand. A companion spreadsheet tool is available upon request.

  13. A model for cell wall dissolution in mating yeast cells: polarized secretion and restricted diffusion of cell wall remodeling enzymes induces local dissolution.

    Directory of Open Access Journals (Sweden)

    Lori B Huberman

    Full Text Available Mating of the budding yeast, Saccharomyces cerevisiae, occurs when two haploid cells of opposite mating types signal using reciprocal pheromones and receptors, grow towards each other, and fuse to form a single diploid cell. To fuse, both cells dissolve their cell walls at the point of contact. This event must be carefully controlled because the osmotic pressure differential between the cytoplasm and extracellular environment causes cells with unprotected plasma membranes to lyse. If the cell wall-degrading enzymes diffuse through the cell wall, their concentration would rise when two cells touched each other, such as when two pheromone-stimulated cells adhere to each other via mating agglutinins. At the surfaces that touch, the enzymes must diffuse laterally through the wall before they can escape into the medium, increasing the time the enzymes spend in the cell wall, and thus raising their concentration at the point of attachment and restricting cell wall dissolution to points where cells touch each other. We tested this hypothesis by studying pheromone treated cells confined between two solid, impermeable surfaces. This confinement increases the frequency of pheromone-induced cell death, and this effect is diminished by reducing the osmotic pressure difference across the cell wall or by deleting putative cell wall glucanases and other genes necessary for efficient cell wall fusion. Our results support the model that pheromone-induced cell death is the result of a contact-driven increase in the local concentration of cell wall remodeling enzymes and suggest that this process plays an important role in regulating cell wall dissolution and fusion in mating cells.

  14. A model for cell wall dissolution in mating yeast cells: polarized se