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Sample records for dramatically improves reconstitution

  1. Improving engraftment and immune reconstitution in umbilical cord blood transplantation

    Directory of Open Access Journals (Sweden)

    Robert eDanby

    2014-02-01

    Full Text Available Umbilical cord blood (UCB is an important source of haematopoietic stem cells (HSC for allogeneic transplantation when HLA-matched sibling and unrelated donors (MUD are unavailable. Although the overall survival rates of UCB transplantation are comparable to the results with MUD, UCB transplants are associated with slow engraftment, delayed immune reconstitution, and increased opportunistic infections. While this may be a consequence of the lower cell dose in UCB grafts, it also reflects the relative immaturity of cellular immunity within cord blood. Furthermore, the limited number of cells and the non-availability of donor lymphocyte infusions (DLI currently prevent the use of post-transplant cellular immunotherapy to boost donor-derived immunity to treat infection, mixed chimerism and disease relapse. Therefore, to further develop UCB transplantation, many strategies to enhance engraftment and immune reconstitution are currently under investigation. This review summarises our current understanding of engraftment and immune recovery following UCB transplantation and why this differs from allogeneic transplants using other sources of HSC. It also provides an comprehensive overview of the promising techniques being used to improve myeloid and lymphoid recovery, including expansion, homing, and delivery of UCB HSC; combined use of UCB with third party donors; isolation and expansion of NK cells, pathogen specific T cells, and regulatory T cells; methods to protect and/or improve thymopoiesis. As many of these strategies are now in clinical trials, it is anticipated that UCB transplantation will continue to advance, further expanding our understanding of UCB biology and HSC transplantation.

  2. Dramatic pretend play games uniquely improve emotional control in young children.

    Science.gov (United States)

    Goldstein, Thalia R; Lerner, Matthew D

    2017-09-15

    Pretense is a naturally occurring, apparently universal activity for typically developing children. Yet its function and effects remain unclear. One theorized possibility is that pretense activities, such as dramatic pretend play games, are a possible causal path to improve children's emotional development. Social and emotional skills, particularly emotional control, are critically important for social development, as well as academic performance and later life success. However, the study of such approaches has been criticized for potential bias and lack of rigor, precluding the ability to make strong causal claims. We conducted a randomized, component control (dismantling) trial of dramatic pretend play games with a low-SES group of 4-year-old children (N = 97) to test whether such practice yields generalized improvements in multiple social and emotional outcomes. We found specific effects of dramatic play games only on emotional self-control. Results suggest that dramatic pretend play games involving physicalizing emotional states and traits, pretending to be animals and human characters, and engaging in pretend scenarios in a small group may improve children's emotional control. These findings have implications for the function of pretense and design of interventions to improve emotional control in typical and atypical populations. Further, they provide support for the unique role of dramatic pretend play games for young children, particularly those from low-income backgrounds. A video abstract of this article can be viewed at: https://youtu.be/2GVNcWKRHPk. © 2017 John Wiley & Sons Ltd.

  3. Critical success factors simplified implementing the powerful drivers of dramatic business improvement

    CERN Document Server

    Howell, Marvin T

    2010-01-01

    Critical-to-success factors (CSFs) have become essential elements to strategic planning and no business can achieve consistent success without effectively adopting them. To take full advantage of CSFs, however, an organization must first understand what they are and how they can be used to drive organizational initiatives and processes. Critical Success Factors Simplified: Implementing the Powerful Drivers of Dramatic Business Improvement provides a concise manual on CSFs that will teach you how to identify and select the right CSFs, measure their impact, and adjust them as needed to reach your goals.

  4. Emulating a crowded intracellular environment in vitro dramatically improves RT-PCR performance

    International Nuclear Information System (INIS)

    Lareu, Ricky R.; Harve, Karthik S.; Raghunath, Michael

    2007-01-01

    The polymerase chain reaction's (PCR) phenomenal success in advancing fields as diverse as Medicine, Agriculture, Conservation, or Paleontology is based on the ability of using isolated prokaryotic thermostable DNA polymerases in vitro to copy DNA irrespective of origin. This process occurs intracellularly and has evolved to function efficiently under crowded conditions, namely in an environment packed with macromolecules. However, current in vitro practice ignores this important biophysical parameter of life. In order to more closely emulate conditions of intracellular biochemistry in vitro we added inert macromolecules into reverse transcription (RT) and PCR. We show dramatic improvements in all parameters of RT-PCR including 8- to 10-fold greater sensitivity, enhanced polymerase processivity, higher specific amplicon yield, greater primer annealing and specificity, and enhanced DNA polymerase thermal stability. The faster and more efficient reaction kinetics was a consequence of the cumulative molecular and thermodynamic effects of the excluded volume effect created by macromolecular crowding

  5. SAFETY IMPROVES DRAMATICALLY IN FLUOR HANFORD SOIL AND GROUNDWATER REMEDIATION PROJECT

    International Nuclear Information System (INIS)

    GERBER MS

    2007-01-01

    This paper describes dramatic improvements in the safety record of the Soil and Groundwater Remediation Project (SGRP) at the Hanford Site in southeast Washington state over the past four years. During a period of enormous growth in project work and scope, contractor Fluor Hanford reduced injuries, accidents, and other safety-related incidents and enhanced a safety culture that earned the SGRP Star Status in the Department of Energy's (DOE's) Voluntary Protection Program (VPP) in 2007. This paper outlines the complex and multi-faceted work of Fluor Hanford's SGRP and details the steps taken by the project's Field Operations and Safety organizations to improve safety. Holding field safety meetings and walkdowns, broadening safety inspections, organizing employee safety councils, intensively flowing down safety requirements to subcontractors, and adopting other methods to achieve remarkable improvement in safety are discussed. The roles of management, labor and subcontractors are detailed. Finally, SGRP's safety improvements are discussed within the context of overall safety enhancements made by Fluor Hanford in the company's 11 years of managing nuclear waste cleanup at the Hanford Site

  6. SAFETY IMPROVES DRAMATICALLY IN FLUOR HANFORD SOIL AND GROUNDWATER REMEDIATION PROJECT

    Energy Technology Data Exchange (ETDEWEB)

    GERBER MS

    2007-12-05

    This paper describes dramatic improvements in the safety record of the Soil and Groundwater Remediation Project (SGRP) at the Hanford Site in southeast Washington state over the past four years. During a period of enormous growth in project work and scope, contractor Fluor Hanford reduced injuries, accidents, and other safety-related incidents and enhanced a safety culture that earned the SGRP Star Status in the Department of Energy's (DOE's) Voluntary Protection Program (VPP) in 2007. This paper outlines the complex and multi-faceted work of Fluor Hanford's SGRP and details the steps taken by the project's Field Operations and Safety organizations to improve safety. Holding field safety meetings and walkdowns, broadening safety inspections, organizing employee safety councils, intensively flowing down safety requirements to subcontractors, and adopting other methods to achieve remarkable improvement in safety are discussed. The roles of management, labor and subcontractors are detailed. Finally, SGRP's safety improvements are discussed within the context of overall safety enhancements made by Fluor Hanford in the company's 11 years of managing nuclear waste cleanup at the Hanford Site.

  7. Use of microsecond current prepulse for dramatic improvements of wire array Z-pinch implosion

    International Nuclear Information System (INIS)

    Calamy, H.; Lassalle, F.; Loyen, A.; Zucchini, F.; Chittenden, J. P.; Hamann, F.; Maury, P.; Georges, A.; Bedoch, J. P.; Morell, A.

    2008-01-01

    The Sphinx machine [F. Lassalle et al., 'Status on the SPHINX machine based on the 1microsecond LTD technology'] based on microsecond linear transformer driver (LTD) technology is used to implode an aluminium wire array with an outer diameter up to 140 mm and maximum current from 3.5 to 5 MA. 700 to 800 ns implosion Z-pinch experiments are performed on this driver essentially with aluminium. Best results obtained before the improvement described in this paper were 1-3 TW radial total power, 100-300 kJ total yield, and 20-30 kJ energy above 1 keV. An auxiliary generator was added to the Sphinx machine in order to allow a multi microsecond current to be injected through the wire array load before the start of the main current. Amplitude and duration of this current prepulse are adjustable, with maxima ∼10 kA and 50 μs. This prepulse dramatically changes the ablation phase leading to an improvement of the axial homogeneity of both the implosion and the final radiating column. Total power was multiplied by a factor of 6, total yield by a factor of 2.5 with a reproducible behavior. This paper presents experimental results, magnetohydrodynamic simulations, and analysis of the effect of such a long current prepulse

  8. Dramatic improvement in genome assembly achieved using doubled-haploid genomes.

    Science.gov (United States)

    Zhang, Hong; Tan, Engkong; Suzuki, Yutaka; Hirose, Yusuke; Kinoshita, Shigeharu; Okano, Hideyuki; Kudoh, Jun; Shimizu, Atsushi; Saito, Kazuyoshi; Watabe, Shugo; Asakawa, Shuichi

    2014-10-27

    Improvement in de novo assembly of large genomes is still to be desired. Here, we improved draft genome sequence quality by employing doubled-haploid individuals. We sequenced wildtype and doubled-haploid Takifugu rubripes genomes, under the same conditions, using the Illumina platform and assembled contigs with SOAPdenovo2. We observed 5.4-fold and 2.6-fold improvement in the sizes of the N50 contig and scaffold of doubled-haploid individuals, respectively, compared to the wildtype, indicating that the use of a doubled-haploid genome aids in accurate genome analysis.

  9. Simple, low-cost ways to dramatically improve the security of tags and seals

    International Nuclear Information System (INIS)

    Johnston, R.G.; Garcia, A.R.E.

    1999-01-01

    The Vulnerability Assessment Team at Los Alamos National Laboratory has analyzed over 100 different tags and security seals (tamper-indicating devices). We have demonstrated how all these security products can be defeated quickly, easily, and inexpensively using low-tech methods. In our view, most of these security devices can be significantly improved with minor changes in their design and/or in how they are used. In this paper, we present some generic suggestions for improving the security and reliability of tags and seals. (author)

  10. Dramatic improvement in photostability of luminescent Eu(III) complexes with tetraphenylimidodiphosphinate ligand

    International Nuclear Information System (INIS)

    Zheng, Wei; Li, Shu-Jing; Li, Cheng-Hui; Zheng, You-Xuan; You, Xiao-Zeng

    2014-01-01

    In this paper, we synthesized and characterized a new Eu(III) tetraphenylimidodiphosphinate complex with dimethyl sulfoxide (DMSO) as co-ligand. Moreover, we compared the photostability of a series of Eu(III) complexes containing tetraphenylimidodiphosphinate ligands, Eu(tpip) 3 , Eu(tpip) 3 Phen, Eu(tpip) 3 DMSO, with their analogs of 1,3-dibenzoylmethanate, Eu(dbm) 3 ∙2H 2 O, Eu(dbm) 3 Phen, Eu(dbm) 3 (DMSO) 2 . We found that the photostability of the luminescent Eu(III) complexes was significantly improved upon substitution of the 1,3-diketones with tetraphenylimidodiphosphinate ligands. -- Highlights: • We synthesized and characterized a new Eu(III) tetraphenylimidodiphosphinate complexes with dimethyl sulfoxide. • We compared the photostability of Eu(III) complex with tetraphenylimidodiphosphinate and 1,3-dibenzoylmethanate ligands. • The photostability is significantly improved in Eu(III) complexes with tetraphenylimidodiphosphinate ligands

  11. Nanocellulose-based Translucent Diffuser for Optoelectronic Device Applications with Dramatic Improvement of Light Coupling.

    Science.gov (United States)

    Wu, Wei; Tassi, Nancy G; Zhu, Hongli; Fang, Zhiqiang; Hu, Liangbing

    2015-12-09

    Nanocellulose is a biogenerated and biorenewable organic material. Using a process based on 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)/NaClO/NaBr system, a highly translucent and light-diffusive film consisting of many layers of nanocellulose fibers and wood pulp microfibers was made. The film demonstrates a combination of large optical transmittance of ∼90% and tunable diffuse transmission of up to ∼78% across the visible and near-infrared spectra. The detailed characterizations of the film indicate the combination of high optical transmittance and haze is due to the film's large packing density and microstructured surface. The superior optical properties make the film a translucent light diffuser and applicable for improving the efficiencies of optoelectronic devices such as thin-film silicon solar cells and organic light-emitting devices.

  12. Dramatic improvement of membrane performance for microalgae harvesting with a simple bubble-generator plate.

    Science.gov (United States)

    Hwang, Taewoon; Oh, You-Kwan; Kim, Bohwa; Han, Jong-In

    2015-06-01

    To overcome fouling issue in membrane-based algae harvesting and thus make an otherwise promising harvesting option more competitive, a bubble-generator plate was developed. According to computational fluid dynamics analysis, the plate generated substantial hydrodynamic power in terms of high pressure, velocity, and shear stress. When installed in a membrane filtration system with membranes of different surface and structural characteristics (one prepared by the phase inversion method, and a commercial one) the bubble-generator was indeed effective in reducing fouling. Without the plate, the much cheaper homemade membrane had the similar performance as the commercial one. Use of the bubble-generator considerably improved the performance of both membranes, and revealed a valuable synergy with the asymmetrical structure of the homemade membrane. This result clearly showed that the ever-problematic fouling could be mitigated in a rather easy manner, and in so doing, that membrane technology could indeed become a practical option for algae harvesting. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Dramatic improvement of crystal quality for low-­temperature-grown rabbit muscle aldolase

    Science.gov (United States)

    Park, HaJeung; Rangarajan, Erumbi S.; Sygusch, Jurgen; Izard, Tina

    2010-01-01

    Rabbit muscle aldolase (RMA) was crystallized in complex with the low-complexity domain (LC4) of sorting nexin 9. Monoclinic crystals were obtained at room temperature that displayed large mosaicity and poor X-ray diffraction. However, orthorhombic RMA–LC4 crystals grown at 277 K under similar conditions exhibited low mosaicity, allowing data collection to 2.2 Å Bragg spacing and structure determination. It was concluded that the improvement of crystal quality as indicated by the higher resolution of the new RMA–LC4 complex crystals was a consequence of the introduction of new lattice contacts at lower temperature. The lattice contacts corresponded to an increased number of interactions between high-entropy side chains that mitigate the lattice strain incurred upon cryocooling and accompanying mosaic spread increases. The thermodynamically unfavorable immobilization of high-entropy side chains used in lattice formation was compensated by an entropic increase in the bulk-solvent content owing to the greater solvent content of the crystal lattice. PMID:20445268

  14. Dramatic improvement of crystal quality for low-temperature-grown rabbit muscle aldolase

    International Nuclear Information System (INIS)

    Park, HaJeung; Rangarajan, Erumbi S.; Sygusch, Jurgen; Izard, Tina

    2010-01-01

    Rabbit muscle aldolase (RMA) was crystallized in complex with the low-complexity domain (LC4) of sorting nexin 9. Monoclinic crystals were obtained at room temperature that displayed large mosaicity and poor X-ray diffraction. However, orthorhombic RMA–LC4 crystals grown at 277 K under similar conditions exhibited low mosaicity, allowing data collection to 2.2 Å Bragg spacing and structure determination. Rabbit muscle aldolase (RMA) was crystallized in complex with the low-complexity domain (LC4) of sorting nexin 9. Monoclinic crystals were obtained at room temperature that displayed large mosaicity and poor X-ray diffraction. However, orthorhombic RMA–LC4 crystals grown at 277 K under similar conditions exhibited low mosaicity, allowing data collection to 2.2 Å Bragg spacing and structure determination. It was concluded that the improvement of crystal quality as indicated by the higher resolution of the new RMA–LC4 complex crystals was a consequence of the introduction of new lattice contacts at lower temperature. The lattice contacts corresponded to an increased number of interactions between high-entropy side chains that mitigate the lattice strain incurred upon cryocooling and accompanying mosaic spread increases. The thermodynamically unfavorable immobilization of high-entropy side chains used in lattice formation was compensated by an entropic increase in the bulk-solvent content owing to the greater solvent content of the crystal lattice

  15. Dramatic Improvement of Crystal Quality for Low-temperature-grown Rabbit Muscle Aldolase

    Energy Technology Data Exchange (ETDEWEB)

    Park, H.; Rangarajan, E; Sygusch, J; Izard, T

    2010-01-01

    Rabbit muscle aldolase (RMA) was crystallized in complex with the low-complexity domain (LC4) of sorting nexin 9. Monoclinic crystals were obtained at room temperature that displayed large mosaicity and poor X-ray diffraction. However, orthorhombic RMA-LC4 crystals grown at 277 K under similar conditions exhibited low mosaicity, allowing data collection to 2.2 {angstrom} Bragg spacing and structure determination. It was concluded that the improvement of crystal quality as indicated by the higher resolution of the new RMA-LC4 complex crystals was a consequence of the introduction of new lattice contacts at lower temperature. The lattice contacts corresponded to an increased number of interactions between high-entropy side chains that mitigate the lattice strain incurred upon cryocooling and accompanying mosaic spread increases. The thermodynamically unfavorable immobilization of high-entropy side chains used in lattice formation was compensated by an entropic increase in the bulk-solvent content owing to the greater solvent content of the crystal lattice.

  16. Dramatic improvement of crystal quality for low-temperature-grown rabbit muscle aldolase.

    Science.gov (United States)

    Park, Hajeung; Rangarajan, Erumbi S; Sygusch, Jurgen; Izard, Tina

    2010-05-01

    Rabbit muscle aldolase (RMA) was crystallized in complex with the low-complexity domain (LC4) of sorting nexin 9. Monoclinic crystals were obtained at room temperature that displayed large mosaicity and poor X-ray diffraction. However, orthorhombic RMA-LC4 crystals grown at 277 K under similar conditions exhibited low mosaicity, allowing data collection to 2.2 A Bragg spacing and structure determination. It was concluded that the improvement of crystal quality as indicated by the higher resolution of the new RMA-LC4 complex crystals was a consequence of the introduction of new lattice contacts at lower temperature. The lattice contacts corresponded to an increased number of interactions between high-entropy side chains that mitigate the lattice strain incurred upon cryocooling and accompanying mosaic spread increases. The thermodynamically unfavorable immobilization of high-entropy side chains used in lattice formation was compensated by an entropic increase in the bulk-solvent content owing to the greater solvent content of the crystal lattice.

  17. Pretransplant mobilization with granulocyte colony-stimulating factor improves B-cell reconstitution by lentiviral vector gene therapy in SCID-X1 mice.

    Science.gov (United States)

    Huston, Marshall W; Riegman, Adriaan R A; Yadak, Rana; van Helsdingen, Yvette; de Boer, Helen; van Til, Niek P; Wagemaker, Gerard

    2014-10-01

    Hematopoietic stem cell (HSC) gene therapy is a demonstrated effective treatment for X-linked severe combined immunodeficiency (SCID-X1), but B-cell reconstitution and function has been deficient in many of the gene therapy treated patients. Cytoreductive preconditioning is known to improve HSC engraftment, but in general it is not considered for SCID-X1 since the poor health of most of these patients at diagnosis and the risk of toxicity preclude the conditioning used in standard bone marrow stem cell transplantation. We hypothesized that mobilization of HSC by granulocyte colony-stimulating factor (G-CSF) should create temporary space in bone marrow niches to improve engraftment and thereby B-cell reconstitution. In the present pilot study supplementing our earlier preclinical evaluation (Huston et al., 2011), Il2rg(-/-) mice pretreated with G-CSF were transplanted with wild-type lineage negative (Lin(-)) cells or Il2rg(-/-) Lin(-) cells transduced with therapeutic IL2RG lentiviral vectors. Mice were monitored for reconstitution of lymphocyte populations, level of donor cell chimerism, and antibody responses as compared to 2 Gy total body irradiation (TBI), previously found effective in promoting B-cell reconstitution. The results demonstrate that G-CSF promotes B-cell reconstitution similar to low-dose TBI and provides proof of principle for an alternative approach to improve efficacy of gene therapy in SCID patients without adverse effects associated with cytoreductive conditioning.

  18. Combining G-CSF with a blockade of adhesion strongly improves the reconstitutive capacity of mobilized hematopoietic progenitor cells.

    Science.gov (United States)

    Christ, O; Kronenwett, R; Haas, R; Zöller, M

    2001-03-01

    Mobilization of hematopoietic progenitor cells is achieved mainly by application of growth factors and, more recently, by blockade of adhesion. In this report, we describe the advantages of a combined treatment with granulocyte colony-stimulating factor (G-CSF) and anti-VLA4 (CD49d)/anti-CD44 as compared to treatment with the individual components. Mobilization by intravenous injection of anti-CD44, anti-VLA4, or G-CSF was controlled in spleen and bone marrow with regard to frequencies of multipotential colony-forming unit (C-CFU), marrow repopulating ability, long-term reconstitution, recovery of myelopoiesis, and regain of immunocompetence. Mobilization by anti-CD44 had a strong effect on expansion of early progenitor cells in the bone marrow, while the recovery in the spleen was poor. In anti-CD49d-mobilized noncommitted and committed progenitors, progenitor expansion was less pronounced, but settlement in the spleen was quite efficient. Thus, anti-CD44 and anti-CD49d differently influenced mobilization. Accordingly, mobilization and recovery after transfer were improved by combining anti-CD44 with anti-CD49d treatment. Mobilization by G-CSF was most efficient with respect to recovery of progenitor cells in the spleen. However, when transferring G-CSF-mobilized cells, regain of immunocompetence was strongly delayed. This disadvantage could be overridden when progenitor cells were mobilized via blockade of adhesion and when expansion of these mobilized progenitor cells was supported by low-dose G-CSF only during the last 24 hours before transfer. Mobilization of pluripotent progenitor cells via antibody blockade of CD44 or CD49d or via G-CSF relies on distinct mechanisms. Therefore, the reconstitutive capacity of a transplant can be significantly improved by mobilization regimens combining antibody with low-dose G-CSF treatment.

  19. Improving reconstituted HDL composition for efficient post-ischemic reduction of ischemia reperfusion injury.

    Directory of Open Access Journals (Sweden)

    Marie-Claude Brulhart-Meynet

    Full Text Available New evidence shows that high density lipoproteins (HDL have protective effects beyond their role in reverse cholesterol transport. Reconstituted HDL (rHDL offer an attractive means of clinically exploiting these novel effects including cardioprotection against ischemia reperfusion injury (IRI. However, basic rHDL composition is limited to apolipoprotein AI (apoAI and phospholipids; addition of bioactive compound may enhance its beneficial effects.The aim of this study was to investigate the role of rHDL in post-ischemic model, and to analyze the potential impact of sphingosine-1-phosphate (S1P in rHDL formulations.The impact of HDL on IRI was investigated using complementary in vivo, ex vivo and in vitro IRI models. Acute post-ischemic treatment with native HDL significantly reduced infarct size and cell death in the ex vivo, isolated heart (Langendorff model and the in vivo model (-48%, p<0.01. Treatment with rHDL of basic formulation (apoAI + phospholipids had a non-significant impact on cell death in vitro and on the infarct size ex vivo and in vivo. In contrast, rHDL containing S1P had a highly significant, protective influence ex vivo, and in vivo (-50%, p<0.01. This impact was comparable with the effects observed with native HDL. Pro-survival signaling proteins, Akt, STAT3 and ERK1/2 were similarly activated by HDL and rHDL containing S1P both in vitro (isolated cardiomyocytes and in vivo.HDL afford protection against IRI in a clinically relevant model (post-ischemia. rHDL is significantly protective if supplemented with S1P. The protective impact of HDL appears to target directly the cardiomyocyte.

  20. Therapeutic doll play in the treatment of a severely impaired psychiatric inpatient: dramatic clinical improvements with a nontraditional nursing intervention.

    Science.gov (United States)

    Birnbaum, Shira; Hanchuk, Hilary; Nelson, Marjorie

    2015-05-01

    Interest has grown in the use of doll therapy, particularly in geropsychiatric and dementia care settings. In a long-term state psychiatric hospital, a dollhouse-play activity was implemented in an effort to engage an acutely disturbed, middle-aged woman undergoing medication trials and whose symptoms had been refractory to conventional treatments. A schedule of nondirective dollhouse-play activities was implemented over an 8-week period. Measures of behavioral change were tracked. Dramatic clinical improvements were seen, including significant reductions in verbal and physical aggression, use of as-needed medications, and need for close one-to-one monitoring. Improvements were seen prior to achievement of therapeutic drug levels. The patient was successfully discharged from the hospital. Doll play has recently been associated with clinical benefits in the care of patients with dementia and has long been deployed in childhood mental health treatment. The current findings suggest doll play may have applications as a time-limited intervention in the treatment of major psychiatric disorders in adults and warrants consideration when achieving therapeutic alliance has proven particularly challenging. Copyright 2015, SLACK Incorporated.

  1. Bariatric Surgery Prior to Total Joint Arthroplasty May Not Provide Dramatic Improvements In Post Arthroplasty Surgical Outcomes

    Science.gov (United States)

    Paxton, Elizabeth W.; Fisher, David; Li, Robert A.; Barber, Thomas C.; Singh, Jasvinder A.

    2014-01-01

    This study compared the total joint arthroplasty (TJA) surgical outcomes of patients who had bariatric surgery prior to TJA to TJA patients who were candidates but did not have bariatric surgery. Patients were retrospectively grouped into: Group 1 (n=69), those with bariatric surgery >2 years prior to TJA, Group 2 (n=102), those with surgery within 2 years of TJA, and Group 3 (n=11,032), those without bariatric surgery. In Group 1, 2.9% (95%CI 0.0–6.9%) had complications within 1 year compared to 5.9% (95%CI 1.3–10.4%) in Group 2, and 4.1% (95%CI 3.8–4.5%) in Group 3. 90-day readmission (7.2%, 95%CI 1.1–13.4%) and revision density (3.4/100 years of observation) was highest in Group 1. Bariatric surgery prior to TJA may not provide dramatic improvements in post-operative TJA surgical outcomes. PMID:24674730

  2. Antifibrotic Therapy in Simian Immunodeficiency Virus Infection Preserves CD4+ T-Cell Populations and Improves Immune Reconstitution With Antiretroviral Therapy

    Science.gov (United States)

    Estes, Jacob D.; Reilly, Cavan; Trubey, Charles M.; Fletcher, Courtney V.; Cory, Theodore J.; Piatak, Michael; Russ, Samuel; Anderson, Jodi; Reimann, Thomas G.; Star, Robert; Smith, Anthony; Tracy, Russell P.; Berglund, Anna; Schmidt, Thomas; Coalter, Vicky; Chertova, Elena; Smedley, Jeremy; Haase, Ashley T.; Lifson, Jeffrey D.; Schacker, Timothy W.

    2015-01-01

    Even with prolonged antiretroviral therapy (ART), many human immunodeficiency virus-infected individuals have <500 CD4+ T cells/µL, and CD4+ T cells in lymphoid tissues remain severely depleted, due in part to fibrosis of the paracortical T-cell zone (TZ) that impairs homeostatic mechanisms required for T-cell survival. We therefore used antifibrotic therapy in simian immunodeficiency virus-infected rhesus macaques to determine whether decreased TZ fibrosis would improve reconstitution of peripheral and lymphoid CD4+ T cells. Treatment with the antifibrotic drug pirfenidone preserved TZ architecture and was associated with significantly larger populations of CD4+ T cells in peripheral blood and lymphoid tissues. Combining pirfenidone with an ART regimen was associated with greater preservation of CD4+ T cells than ART alone and was also associated with higher pirfenidone concentrations. These data support a potential role for antifibrotic drug treatment as adjunctive therapy with ART to improve immune reconstitution. PMID:25246534

  3. GABA dramatically improves glucose tolerance in streptozotocin-induced diabetic rats fed with high-fat diet.

    Science.gov (United States)

    Sohrabipour, Shahla; Sharifi, Mohammad Reza; Talebi, Ardeshir; Sharifi, Mohammadreza; Soltani, Nepton

    2018-05-05

    Skeletal muscle, hepatic insulin resistance, and beta cell dysfunction are the characteristic pathophysiological features of type 2 diabetes mellitus. GABA has an important role in pancreatic islet cells. The present study attempted to clarify the possible mechanism of GABA to improve glucose tolerance in a model of type 2 diabetes mellitus in rats. Fifty Wistar rats were divided into five groups: NDC that was fed the normal diet, CD which received a high-fat diet with streptozotocin, CD-GABA animals that received GABA via intraperitoneal injection, plus CD-Ins1 and CD-Ins2 groups which were treated with low and high doses of insulin, respectively. Body weight and blood glucose were measured weekly. Intraperitoneal glucose tolerance test (IPGTT), insulin tolerance test (ITT), urine volume, amount of water drinking, and food intake assessments were performed monthly. The hyperinsulinemic euglycemic clamp was done for assessing insulin resistance. Plasma insulin and glucagon were measured. Abdominal fat was measured. Glucagon receptor, Glucose 6 phosphatase, Phosphoenolpyruvate carboxykinase genes expression were evaluated in liver and Glucose transporter 4 (GLUT4) genes expression and protein translocation were evaluated in the muscle. GABA or insulin therapy improved blood glucose, insulin level, IPGTT, ITT, gluconeogenesis pathway, Glucagon receptor, body weight and body fat in diabetic rats. GLUT4 gene and protein expression increased. GABA whose beneficial effect was comparable to that of insulin, also increased glucose infusion rate during an euglycemic clamp. GABA could improve insulin resistance via rising GLUT4 and also decreasing the gluconeogenesis pathway and Glucagon receptor gene expression. Copyright © 2018. Published by Elsevier B.V.

  4. Intravenous immunoglobulin therapy leading to dramatic improvement in a patient with systemic juvenile idiopathic arthritis and severe pericarditis resistant to steroid pulse therapy.

    Science.gov (United States)

    Aizawa-Yashiro, Tomomi; Oki, Eishin; Tsuruga, Kazushi; Nakahata, Tohru; Ito, Etsuro; Tanaka, Hiroshi

    2012-05-01

    A 7-year-old Japanese boy with a 4-month history of systemic juvenile idiopathic arthritis (s-JIA) experienced disease flare with spiking fever, exanthema and arthralgia. He then developed progressive dyspnea due to severe pericarditis, and proinflammatory hypercytokinemia was suspected. Methylprednisolone pulse therapy was ineffective and echocardiography showed massive pericardial effusion had persisted. Alternatively, subsequent intravenous immunoglobulin (IVIG) therapy resulted in dramatic resolution of the pericardial effusion, and his general condition significantly improved within a few days. This case report may lend further support the use of IVIG for selected patients with s-JIA and severe pericarditis.

  5. Creative Dramatics. Beginnings Workshop.

    Science.gov (United States)

    Gabriel, Julia; Sidlovskaya, Olga; Stotter, Ruth; Haugen, Kirsten; Leithold, Naomi

    2000-01-01

    Presents five articles on using creative dramatics in early childhood education: (1) "Drama: A Rehearsal for Life" (Julia Gabriel); (2) "Fairy Tales Enhance Imagination and Creative Thinking" (Olga Sidlovskaya); (3) "Starting with a Story" (Ruth Stotter); (4) "Using Creative Dramatics to Include All…

  6. Reconstitution of halorhodopsin

    Energy Technology Data Exchange (ETDEWEB)

    Kong, T.

    1989-11-01

    Halobacterium halobium contains a family of retinal-bound proteins: bacteriorhodopsin (bR) which mediates phototrophic growth as a light-riven proton pump, halorhodopsin (hR) which is a light-driven chloride pump, and one or more sensory rhodopsins (sR) which mediate a phototactic response. Two-dimensional crystallization of halorhodopsin has been attempted though the reconstitution of purified halorhodopsin with purple membrane lipid for electron microscopy work. The first important step for crystallization is to get a homogeneous protein which is pure and not denatured. Homogeneous halorhodopsin has been obtained by a modification of existing purification methods. Some nice looking membrane patches which have the same density as purple membrane have been obtained. But unfortunately, they are not crystalline. The procedure of hR reconstitution is described in detail and some other strategies to induce the protein crystal in the reconstituted membrane are discussed in this dissertation. 76 refs., 20 figs., 6 tabs.

  7. Lord Byron's dramatic work

    Directory of Open Access Journals (Sweden)

    Andrejević Ana

    2009-01-01

    Full Text Available All of Byron's six finished dramas aspire to display conflict between the individual will and the strength of the universal and omnipresent inevitability. Although he accentuated classical dramatic rules or three dramatic unities as the way to construct a good drama, he couldn't escape from the deep rooted romantic feeling of his genius. His plays are thus pictures of existing existential revolt with the protagonists true to their feelings, always consistent, active and devoted to one general idea. They are estranged from the meaningless world that leads them to rebellion and death, but prometheously constructed are the most successful and memorable element of his plays. Byron as a dramatist always tries to find an everlasting dimension and virtue in the meaningless world through the construction of personal myth and thus the main subject of all his dramas is freedom in all its aspects.

  8. Poly(2,5-dimercapto-1,3,4-thiadiazole) as a Cathode for Rechargeable Lithium Batteries with Dramatically Improved Performance

    KAUST Repository

    Gao, Jie

    2012-05-29

    Organosulfur compounds with multiple thiol groups are promising for high gravimetric energy density electrochemical energy storage. We have synthesized a poly(2,5-dimercapto-1,3,4-thiadiazole) (PDMcT)/poly(3,4-ethylenedioxythiophene) (PEDOT) composite cathode for lithium-ion batteries with a new method and investigated its electrochemical behavior by charge/discharge cycles and cyclic voltammetry (CV) in an ether-based electrolyte. Based on a comparison of the electrochemical performance with a carbonate-based electrolyte, we found a much higher discharge capacity, but also a very attractive cycling performance of PDMcT by using a tetra(ethylene glycol) dimethyl ether (TEGDME)-based electrolyte. The first discharge capacity of the as-synthesized PDMcT/PEDOT composite approached 210 mAh g -1 in the TEGDME-based electrolyte. CV results clearly show that the redox reactions of PDMcT are highly reversible in this TEGDME-based electrolyte. The reversible capacity remained around 120 mAh g -1 after 20 charge/discharge cycles. With improved cycling performance and very low cost, PDMcT could become a very promising cathode material when combined with a TEGDME-based electrolyte. The poor capacity in the carbonate-based electrolyte is a consequence of the irreversible reaction of the DMcT monomer and dimer with the solvent, emphasizing the importance of electrolyte chemistry when studying molecular-based battery materials. The nature of the electrolyte has a dramatic effect on the performance of poly(2,5-dimercapto-1,3,4-thiadiazole) (PDMcT) as a cathode material in lithium-ion batteries. Whereas the use of an ethylene/diethyl carbonate (EC/DEC)-based electrolyte resulted in very poor performance, the use of a tetra(ethylene glycol) dimethyl ether (TEGDME)-based electrolyte dramatically improved the performance in terms of both the discharge capacity and capacity retention (see scheme). Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Reconstituted products from oak

    Science.gov (United States)

    W. C. Lewis; B. G. Heebink

    1971-01-01

    "Reconstituted" describes a family of panel products made from fractionated oak, bonded with either a synthetic resin or a natural lignin bond. Several current commercial fiber panel products from oak are described, and the status of research on experimental products and processes is presented. Recent technological developments are removing the stigma...

  10. The Reconstituted Family

    OpenAIRE

    Talbot, Yves

    1981-01-01

    The reconstituted or step-family is becoming more prevalent. The physician who cares for families should be acquainted with the different aspects of such family structure and family functioning. This will enable professionals to better understand and assist their patients, by anticipating the different stresses related to the new family formation, and supporting their adaptation.

  11. Transplantation of motoneurons derived from MASH1-transfected mouse ES cells reconstitutes neural networks and improves motor function in hemiplegic mice.

    Science.gov (United States)

    Ikeda, Ritsuko; Kurokawa, Manae S; Chiba, Shunmei; Yoshikawa, Hideshi; Hashimoto, Takuo; Tadokoro, Mamoru; Suzuki, Noboru

    2004-10-01

    Mouse embryonic stem (ES) cells were transfected with a MASH1 expression vector and G418-resistant cells were selected. The MASH1-transfected cells became neuron-like appearance and expressed betaIIItubulin and panNCAM. Glial fibrillary acidic protein (GFAP) and galactocerebroside (GalC)-expressing cells were rarely detected. Half of the neural cells differentiated into the Islet1+ motoneuron lineage. Thus, we obtained motoneuron lineage-enriched neuronal cells by transfection of ES cells with MASH1. A hemiplegic model of mice was developed by cryogenic injury of the motor cortex, and motoneuron lineage-enriched neuronal cells were transplanted underneath the injured motor cortex neighboring the periventricular region. The motor function of the recipients was assessed by a beam walking and rotarod tests, whereby the results gradually improved, but little improvement was observed in vehicle injected control mice. We found that the grafted cells not only remained close to the implantation site, but also exhibited substantial migration, penetrating into the damaged lesion in a directed manner up to the cortical region. Grafted neuronal cells that had migrated into the cortex were elongated axon-positive for neurofilament middle chain (NFM). Synaptophysin immunostaining showed a positive staining pattern around the graft, suggesting that the transplanted neurons interacted with the recipient neurons to form a neural network. Our study suggests that the motoneuron lineage can be induced from ES cells, and grafted cells adapt to the host environment and can reconstitute a neural network to improve motor function of a paralyzed limb.

  12. Fuel assembly reconstitution

    International Nuclear Information System (INIS)

    Morgado, Mario M.; Oliveira, Monica G.N.; Ferreira Junior, Decio B.M.; Santos, Barbara O. dos; Santos, Jorge E. dos

    2009-01-01

    Fuel failures have been happened in Nuclear Power Plants worldwide, without lost of integrity and safety, mainly for the public, environment and power plants workers. The most common causes of these events are corrosion (CRUD), fretting and pellet cladding interaction. These failures are identified by increasing the activity of fission products, verified by chemical analyses of reactor coolant. Through these analyses, during the fourth operation cycle of Angra 2 Nuclear Power Plant, was possible to observe fuel failure indication. This indication was confirmed in the end of the cycle during the unloading of reactor core through leakage tests of fuel assembly, using the equipment called 'In Mast Sipping' and 'Box Sipping'. After confirmed, the fuel assembly reconstitution was scheduled, and happened in April, 2007, where was identified the cause and the fuel rod failure, which was substitute by dummy rods (zircaloy). The cause was fretting by 'debris'. The actions to avoid and prevent fuel assemblies failures are important. The goals of this work are to describe the methodology of fuel assembly reconstitution using the FARE (Fuel Assembly Reconstitution Equipment) system, to describe the results of this task in economic and security factors of the company and show how the fuel assembly failures are identified during operation and during the outage. (author)

  13. Idiopathic Facial Aseptic Granuloma in a 13-Year-Old Boy Dramatically Improved with Oral Doxycycline and Topical Metronidazole: Evidence for a Link with Childhood Rosacea

    Directory of Open Access Journals (Sweden)

    Camille Orion

    2016-07-01

    Full Text Available Idiopathic facial aseptic granuloma (IFAG is a rare, benign pediatric dermatological lesion that occurs in children between 8 months and 13 years of age. The pathogenesis of IFAG is still unclear but it is likely to be associated with granulomatous rosacea in childhood. Here we describe a case of IFAG in a 13-year-old boy who showed a dramatic response to oral doxycycline and topical metronidazole, which supports the hypothesis that IFAG may belong to the spectrum of rosacea.

  14. An Analysis of the Impact of Deregulating the Pricing of Reconstituted Milk Under Federal Milk Marketing Orders

    OpenAIRE

    Novakovic, Andrew M.; Aplin, Richard D.

    1982-01-01

    This study reports an effort to improve estimates of the cost of reconstituted milk relative to fresh milk. The cost advantage of reconstituted milk is calculated for representative plants in six cities across the U.S. The implications for farm prices of lower cost reconstituted milk are discussed.

  15. Primary Care DirectConnect: How the Marriage of Call Center Technology and the EMR Brought Dramatic Results—A Service Quality Improvement Study

    Science.gov (United States)

    Bowman, Brent; Smith, Scott

    2010-01-01

    Of the key Health Plan patient satisfaction measures used in Kaiser Permanente Colorado, ease of contacting the physician's office with a medical question was consistently rated as the lowest quarterly patient satisfaction measure. Furthermore, medical office staff had become dissatisfied with their inability to contact patients who had previously left messages. In addition to the shear volume of messages, the return calls were often unanswered, leading to subsequent attempts to reach patients, creating additional work for medical office staff. DirectConnect—the project name for a system and set of processes focused on improving patient satisfaction with the ability to contact Primary Care delivery teams by telephone—focuses on isolating medical advice calls from the other types of calls handled by the centralized Call Center. The system identifies the patient using his/her unique electronic medical record number, then automatically routes medical advice calls directly to the appropriate Primary Care Physician (PCP) or staff. The clinician may then evaluate and respond to the patient's need quickly, thus managing more of their panel's requests in real time. How is DirectConnect different from simply having the patient contact their PCP's office directly? The primary difference is “one-number” convenience that allows all patients to dial one number to access their PCP's team. In addition, calls are routed to various staff as available to reduce long telephone queues and wait times. The DirectConnect system has resulted in statistically significant improvement in key service quality measures. Patient satisfaction improved from a pre-implementation nine quarter mean of 55.9% to a post-implementation 12 quarter mean of 70.2%. Fourteen percent to 17% of all Primary Care calls are now handled by the patient's home medical office team, creating a 54% improvement in the centralized Call Center's speed of answering calls in the first quarter post implementation

  16. Dramatically improve the Safety Performance of Li ion Battery Separators and Reduce the Manufacturing Cost Using Ultraviolet Curing and High Precision Coating Technologies

    Energy Technology Data Exchange (ETDEWEB)

    Voelker, Gary [Miltec UV International, LLC, Stevensville, MD (United States); Arnold, John [Miltec UV International, LLC, Stevensville, MD (United States)

    2017-06-30

    The objective of this project was to improve the safety of operation of Lithium ion batteries (LIB)and at the same time significantly reduce the manufacturing cost of LIB separators. The project was very successful in demonstrating the improved performance and reduced cost attributed to using UV curable binder and high speed printing technology to place a very thin and precisely controlled ceramic layer on the surface of base separators made of polyolefins such as Polyethylene, Polypropylene and combinations of the two as well as cellulosic base separators. The underlying need for this new technology is the recently identified potential of fire in large format Lithium ion batteries used in hybrid, plug-in hybrid and electric vehicles. The primary potential cause of battery fire is thermal runaway caused by several different electrical or mechanical mechanisms; such as, overcharge, puncture, overheating, compaction, and internal short circuit. During thermal runaway, the ideal separator prevents ion flow and continues to physically separate the anode from the cathode. If the temperature of the battery gets higher, the separator may melt and partially clog the pores and help prevent ion flows but it also can shrink which can result in physical contact of the electrodes and accelerate thermal run-away even further. Ceramic coated separators eliminate many of the problems related to the usage of traditional separators. The ceramic coating provides an electrically insulating layer that retains its physical integrity at high temperature, allows for more efficient thermal heat transfer, helps reduce thermal shrinkage, and inhibits dendrite growth that could create a potential short circuit. The use of Ultraviolet (UV) chemistry to bind fine ceramic particles on separators is a unique and innovative approach primarily because of the instant curing of the UV curable binder upon exposure to UV light. This significant reduction in drying/curing time significantly reduces the

  17. Surface modification of alignment layer by ultraviolet irradiation to dramatically improve the detection limit of liquid-crystal-based immunoassay for the cancer biomarker CA125.

    Science.gov (United States)

    Su, Hui-Wen; Lee, Mon-Juan; Lee, Wei

    2015-05-01

    Liquid crystal (LC)-based biosensing has attracted much attention in recent years. We focus on improving the detection limit of LC-based immunoassay techniques by surface modification of the surfactant alignment layer consisting of dimethyloctadecyl[3-(trimethoxysilyl)propyl]ammonium chloride (DMOAP). The cancer biomarker CA125 was detected with an array of anti-CA125 antibodies immobilized on the ultraviolet (UV)-modified DMOAP monolayer. Compared with a pristine counterpart, UV irradiation enhanced the binding affinity of the CA125 antibody and reproducibility of immunodetection in which a detection limit of 0.01 ng∕ml for the cancer biomarker CA125 was achieved. Additionally, the optical texture observed under a crossed polarized microscope was correlated with the analyte concentration. In a proof-of-concept experiment using CA125-spiked human serum as the analyte, specific binding between the CA125 antigen and the anti-CA125 antibody resulted in a distinct and concentration-dependent optical response despite the high background caused by nonspecific binding of other biomolecules in the human serum. Results from this study indicate that UVmodification of the alignment layer, as well as detection with LCs of large birefringence, contributes to the enhanced performance of the label-free LC-based immunodetection, which may be considered a promising alternative to conventional label-based methods.

  18. Health Sensors, Smart Home Devices, and the Internet of Medical Things: An Opportunity for Dramatic Improvement in Care for the Lower Extremity Complications of Diabetes.

    Science.gov (United States)

    Basatneh, Rami; Najafi, Bijan; Armstrong, David G

    2018-05-01

    The prevalent and long-neglected diabetic foot ulcer (DFU) and its related complications rank among the most debilitating and costly sequelae of diabetes. With the rise of the Internet of medical things (IoMT), along with smart devices, the med-tech industry is on the cusp of a home-care revolution, which could also create opportunity for developing effective solutions with significant potential to reduce DFU-associated costs and saving limbs. This article discusses potential applications of IoMT to the DFU patient population and beyond. To better understand potential opportunities and challenges associated with implementing IoMT for management of DFU, the authors reviewed recent relevant literatures and included their own expert opinions from a multidisciplinary point of view including podiatry, engineering, and data security. The IoMT has opened digital transformation of home-based diabetic foot care, as it enables promoting patient engagement, personalized care and smart management of chronic and noncommunicable diseases through individual data-driven treatment regimens, telecommunication, data mining, and comprehensive feedback tailored to individual requirements. In particular, with recent advances in voice-activated commands technology and its integration as a part of IoMT, new opportunities have emerged to improve the patient's central role and responsibility in enabling an optimized health care ecosystem. The IoMT has opened new opportunities in health care from remote monitoring to smart sensors and medical device integration. While it is at its early stage of development, ultimately we envisage a connected home that, using voice-controlled technology and Bluetooth-radio-connected add-ons, may augment much of what home health does today.

  19. La dramatúrgia docent

    OpenAIRE

    Comelles, Salvador

    2009-01-01

    La vida és un teatre. Una dita que lluny de semblar despectiva, resumeix la realitat humana diària. Perquè la vida és dramatúrgia. Un concepte menys agressiu que evoca les diferents situacions comunicatives que les persones enfronten al llarg de la seva vida per tal de complir els seus objectius. I ho fan assumint rols i actituds que una situació concreta requereix, com ara, la vida del docent i l’àmbit professional i social on es desenvolupa. Identificar l’ombra dramatúrgica que és desprèn d...

  20. Learning through Dramatic Story Presentation

    Science.gov (United States)

    Tindall, Evie

    2012-01-01

    The use of story with dramatic presentation approaches produces an engaging and powerful instructional choice for today's adult ESL educators. Two engaging and timed-tested approaches are Reader's Theater and Tableau Vivant. Both provide English language learners with content tailored to their abilities in addition to numerable opportunities to…

  1. Little Eyolf and dramatic tradition

    Directory of Open Access Journals (Sweden)

    Roland Lysell

    2015-02-01

    Full Text Available The article criticises an Ibsen tradition who has seen the last scene of Little Eyolf as a reconciliation. Instead, the article discusses the improbability of a happy marriage characterised by social engagement. The play is open but it is hardly probable that Rita, with her erotic desire, and Allmers, whose desire has turned into metaphysics, can be happy together. The arguments refer to inner criteria and the constantly present dramatic tradition.

  2. Dramatic weight loss associated with commencing clozapine

    OpenAIRE

    Lally, John; McDonald, Colm

    2011-01-01

    The authors report the case of a 44-year-old man with a long history of chronic enduring schizophrenia who experienced dramatic weight loss after commencing treatment with clozapine, an antipsychotic medication characteristically associated with the greatest degree of weight gain among medical treatments for schizophrenia. He was obese with a body mass index (BMI) of 41.5 kg/m2, but after commencing clozapine therapy he experienced an improvement in psychotic symptoms and 40% loss of his body...

  3. Reconstitution of an efficient thymidine salvage pathway in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Vernis, L.; Piskur, Jure; Diffley, J.F.X.

    2003-01-01

    The budding yeast Saccharomyces cerevisiae is unable to incorporate exogenous nucleosides into DNA. We have made a number of improvements to existing strategies to reconstitute an efficient thymidine salvage pathway in yeast. We have constructed strains that express both a nucleoside kinase as well...

  4. Effect of specialized combined strains on reconstituted milk reduced ...

    African Journals Online (AJOL)

    On the other hand, results of the main flavor compounds in CRMP indicated that combination of the EPS-producing S. thermophilus TM11 and an adjunct Lactococcus lactis ssp. lactis could provide CRMP with novel and proper flavor properties while improving the textural characteristics. Key words: Reconstituted milk, ...

  5. Configuration management after design basis reconstitution

    International Nuclear Information System (INIS)

    Purcell, J.J.; Livingston, B.R.

    1991-01-01

    Over the last few years, Fort Calhoun station (FCS) has implemented a number of programs to enhance plant operability and readiness. The design basis document (DBD) reconstitution project was the cornerstone of this effort. Vendor manual upgrade, operating procedures upgrade, plant equipment data-base verification, equipment labeling, and warehousing improvements were also implemented as part of this improvement program. With the completion of these programs, plant documentation was current to the baselines established by each program, and a configuration management program (CMP) was established to maintain this level of accuracy throughout the remaining life of FCS. Change control throughout the organization has been reviewed and upgraded to ensure that all changes are evaluated for impact to the design bases

  6. Reconstitutable control rod spider assembly

    International Nuclear Information System (INIS)

    Shallenberger, J.M.; Ferian, S.J.

    1990-01-01

    A reconstitutable control rod/spider assembly includes a hollow connecting finger of the spider having a pair of opposing flat segments formed on the interior thereof and engaging a pair of opposing flat sectors formed on the exterior of a stem extending form the upper end of control rod. The stem also has an externally-threaded portion engaging a nut and a pilot aligning portion for the nut. The nut has a radially flexible and expandable thread-defining element captured in its bore. The segments and sectors allow the rod to be removed and reattached after turning through 180 0 to allow more even wear on the rod. (author)

  7. Dramatic weight loss associated with commencing clozapine.

    Science.gov (United States)

    Lally, John; McDonald, Colm

    2011-11-08

    The authors report the case of a 44-year-old man with a long history of chronic enduring schizophrenia who experienced dramatic weight loss after commencing treatment with clozapine, an antipsychotic medication characteristically associated with the greatest degree of weight gain among medical treatments for schizophrenia. He was obese with a body mass index (BMI) of 41.5 kg/m(2), but after commencing clozapine therapy he experienced an improvement in psychotic symptoms and 40% loss of his body weight attained through an altered diet and exercise regime, which resulted in him attaining a normal BMI of 24.8 kg/m(2).

  8. Increasing Stem Cell Dose Promotes Posttransplant Immune Reconstitution.

    Science.gov (United States)

    Xu, Ning; Shen, Sylvie; Dolnikov, Alla

    2017-04-01

    Umbilical cord blood (UCB) transplantation can provide a successful therapeutic option for patients that have no suitable related donor. UCB transplantation is often limited by the relatively small hematopoietic stem cell (HSC) numbers in UCB especially for adult recipients. Early neutrophil and platelet engraftment correlates with the stem cell numbers in UCB transplant. Compared to other HSC sources, immune reconstitution following UCB transplant is slower and complicated by increased frequency of opportunistic infections. The effect of HSC numbers in UCB transplant on immune reconstitution was not thoroughly examined. Using immunocompromised mice transplanted with purified UCB CD34+ stem cells, we have demonstrated that increasing the numbers of CD34+ cells in the transplant promotes hematopoietic and immune reconstitution. At early stages posttransplant, high stem cell dose generated relatively more B cells, while lower dose generated more myeloid and T cells. Thus, the size of the stem cell graft appears to modulate the differentiation potential of infused stem cells. In addition, increasing stem cell dose in the transplant improved CD8+ T cell development and delayed late memory T cell skewing in expense of naive T cells highlighting the importance of HSC dose to maintain the pool of naive T cells able to develop strong immune responses. Transplantation of ex vivo expanded CD34+ cells did not promote, but rather delayed immune reconstitution suggesting the loss of primitive lymphoid precursor cells during ex vivo expansion.

  9. Reconstitution of fuel assemblies and core components

    Energy Technology Data Exchange (ETDEWEB)

    Hummel, Wolfgang; Langenberger, Jan [AREVA NP GmbH (Germany)

    2012-11-01

    Due to AREVA's experience and big portfolio of techniques, reconstitution of fuel assemblies and core components at light water reactors is possible within a reasonable timeframe and with interesting cost benefit. Customer feedback indicates the sustainability of such reconstitutions. As a result, a long-term maintenance of value can be assured and early waste disposal can be avoided. (orig.)

  10. Altered parenting and the reconstituted family.

    Science.gov (United States)

    Whitley, G G; Kachel, J M

    1991-01-01

    Reconstituted families are a high incidence phenomena in contemporary society. Because the family continues to be a focus for the delivery of nursing care, nurses must now address the special needs of individuals who are members of reconstituted families. Studies in this area provide important background information regarding behavioral patterns in reconstituted families that can be used for assessment and intervention with these families. Through the use of focused assessment parameters, nurses can collect data that will indicate the special needs of members of reconstituted families. In general, nursing interventions with clients who are a part of a reconstituted family fall into two major categories: (a) developing positive parenting behaviors, and (b) protecting the development of the stepchild.

  11. Recent advances on Charpy specimen reconstitution techniques

    Energy Technology Data Exchange (ETDEWEB)

    Andrade, Arnaldo H.P.; Lobo, Raquel M.; Miranda, Carlos Alexandre J., E-mail: aandrade@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2017-07-01

    Charpy specimen reconstitution is widely used around the world as a tool to enhance or supplement surveillance programs of nuclear reactor pressure vessels. The reconstitution technique consists in the incorporation of a small piece from a previously tested specimen into a compound specimen, allowing to increase the number of tests. This is especially important if the available materials is restricted and fracture mechanics parameter have to be determined. The reconstitution technique must fulfill some demands, among them tests results like the original standard specimens and the loaded material of the insert must not be influenced by the welding and machining procedure. It is known that reconstitution of Charpy specimens may affect the impact energy in a consequence of the constraint of plastic deformation by the hardened weldment and HAZ. This paper reviews some recent advances of the reconstitution technique and its applications. (author)

  12. Recent advances on Charpy specimen reconstitution techniques

    International Nuclear Information System (INIS)

    Andrade, Arnaldo H.P.; Lobo, Raquel M.; Miranda, Carlos Alexandre J.

    2017-01-01

    Charpy specimen reconstitution is widely used around the world as a tool to enhance or supplement surveillance programs of nuclear reactor pressure vessels. The reconstitution technique consists in the incorporation of a small piece from a previously tested specimen into a compound specimen, allowing to increase the number of tests. This is especially important if the available materials is restricted and fracture mechanics parameter have to be determined. The reconstitution technique must fulfill some demands, among them tests results like the original standard specimens and the loaded material of the insert must not be influenced by the welding and machining procedure. It is known that reconstitution of Charpy specimens may affect the impact energy in a consequence of the constraint of plastic deformation by the hardened weldment and HAZ. This paper reviews some recent advances of the reconstitution technique and its applications. (author)

  13. Directed diffusion of reconstituting dimers

    International Nuclear Information System (INIS)

    Barma, Mustansir; Grynberg, Marcelo D; Stinchcombe, Robin B

    2007-01-01

    We discuss the dynamical aspects of an asymmetric version of assisted diffusion of hard core particles on a ring studied by Menon et al (1997 J. Stat. Phys. 86 1237). The asymmetry brings in phenomena like kinematic waves and effects of the Kardar-Parisi-Zhang non-linearity, which combine with the feature of strongly broken ergodicity, a characteristic of the model. A central role is played by a single non-local invariant, the irreducible string, whose interplay with the driven motion of reconstituting dimers, arising from the assisted hopping, determines the asymptotic dynamics and scaling regimes. These are investigated both analytically and numerically through sector-dependent mappings to the asymmetric simple exclusion process

  14. Directed diffusion of reconstituting dimers

    Energy Technology Data Exchange (ETDEWEB)

    Barma, Mustansir [Department of Theoretical Physics, Tata Institute of Fundamental Research, Mumbai 400005 (India); Grynberg, Marcelo D [Departamento de Fisica, Universidad Nacional de La Plata (1900) La Plata (Argentina); Stinchcombe, Robin B [Isaac Newton Institute for Mathematical Sciences, 20 Clarkson Road, Cambridge CB3 0EH (United Kingdom)

    2007-02-14

    We discuss the dynamical aspects of an asymmetric version of assisted diffusion of hard core particles on a ring studied by Menon et al (1997 J. Stat. Phys. 86 1237). The asymmetry brings in phenomena like kinematic waves and effects of the Kardar-Parisi-Zhang non-linearity, which combine with the feature of strongly broken ergodicity, a characteristic of the model. A central role is played by a single non-local invariant, the irreducible string, whose interplay with the driven motion of reconstituting dimers, arising from the assisted hopping, determines the asymptotic dynamics and scaling regimes. These are investigated both analytically and numerically through sector-dependent mappings to the asymmetric simple exclusion process.

  15. Development of Reconstitution Technology for Surveillance Specimens

    International Nuclear Information System (INIS)

    Yasushi Atago; Shunichi Hatano; Eiichiro Otsuka

    2002-01-01

    The Japan Power Engineering and Inspection Corporation (JAPEIC) has been carrying out the project titled 'Nuclear Power Plant Integrated Management Technology (PLIM)' consigned by Japanese Ministry of Economy, Trade and Industry (METI) since 1996FY as a 10-years project. As one of the project themes, development of reconstitution technology for reactor pressure vessel (RPV/RV) surveillance specimens, which are installed in RPVs to monitor the neutron irradiation embrittlement on RPV/RV materials, is now on being carried out to deal with the long-term operation of nuclear power plants. The target of this theme is to establish the technical standard for applicability of reconstituted surveillance specimens including the reconstitution of the Charpy specimens and Compact Tension (CT) specimens. With the Charpy specimen reconstitution, application of 10 mm length inserts is used, which enables the conversion of tests from the LT-direction to the TL-direction. This paper presents the basic data from Charpy and CT specimens of RPV materials using the surveillance specimens obtained for un-irradiated materials including the following. 1) Reconstitution Technology of Charpy Specimens. a) The interaction between plastic zone and Heat Affected Zone (HAZ). b) The effects of the possible deviations from the standard specimens for the reconstituted specimens. 2) Reconstitution Technology of CT specimens. a) The correlation between fracture toughness and plastic zone width. Because the project is now in progress, this paper describes the outline of the results obtained as of the end of 2000 FY. (authors)

  16. Influence of bromide on the performance of the amphipod Hyalella azteca in reconstituted waters

    Science.gov (United States)

    Ivey, Chris D.; Ingersoll, Christopher G.

    2016-01-01

    Poor performance of the amphipod Hyalella azteca has been observed in exposures using reconstituted waters. Previous studies have reported success in H. azteca water-only exposures with the addition of relatively high concentrations of bromide. The present study evaluated the influence of lower environmentally representative concentrations of bromide on the response ofH. azteca in 42-d water-only exposures. Improved performance of H. azteca was observed in reconstituted waters with >0.02 mg Br/L.

  17. Framatome experience in fuel assembly repair and reconstitution

    International Nuclear Information System (INIS)

    Leroy, G.

    1998-01-01

    Since 1985, FRAMATOME has build up extensive experience in the poolside replacement of fuel rods for repair or R and D purposes and the reconstitution of fuel assemblies (i.e. replacement of a damaged structure to enable reuse of the fuel rod bundle). This experience feedback enables FRAMATOME to improve in steps the technical process and the equipment used for the above operations in order to enhance their performance in terms of setup, flexibility, operating time and safety. In parallel, the fuel assembly and fuel rod designs have been modified to meet the same goals. The paper will describe: - the overall experience of FRAMATOME with UO 2 fuel as well as MOX fuel; the usual technical process used for fuel replacement and the corresponding equipment set; - the usual technical process for fuel assembly reconstitution and the corresponding equipment set. This process is rather unique since it takes profit of the specific FRAMATOME fuel assembly design with removable top and bottom nozzles, so that fuel rods insertion by pulling through in the new structure is similar to what is done in the manufacturing plant; - the usual inspections done on the fuel rods and/or the fuel assembly; - the design of the new reconstitution equipment (STAR) compared with the previous one as well as their comparative performance. The final section will be a description of the alternative reconstitution process and equipment used by FRAMATOME in reactors in which the process cannot be used for several reasons such as compatibility or administrative authorization. This process involves the pushing of fuel rods into the new structure, requiring further precautions. (author)

  18. Functional reconstitution of the glycine receptor

    Energy Technology Data Exchange (ETDEWEB)

    Garcia-Calvo, M.; Ruiz-Gomez, A.; Vazquez, J.; Morato, E.; Valdivieso, F.; Mayor, F. Jr. (Universidad Autonoma de Madrid (Spain))

    1989-07-25

    The functional reconstitution of the chloride channel coupled glycine receptor is described. Glycine receptors were purified from the cholate extract of rat spinal cord membranes by affinity chromatography and incorporated into phospholipid vesicles by the addition of phosphatidylcholine and removal of detergent by gel filtration. The reconstituted vesicles showed the same polypeptide composition as the purified receptor. The pharmacological characteristics of the glycine receptor were also preserved in the proteoliposomes, as demonstrated by the displacement of ({sup 3}H)strychnine binding by several glycinergic ligands and by photoaffinity labeling experiments. In order to observe functional responses (i.e., specific agonist-induced anion translocation), the authors have developed an assay based on the fluorescence quenching of an anion-sensitive entrapped probe, SPQ (6-methoxy-N-(3-sulfopropyl)quinolinium). Reconstituted vesicles were loaded with the fluorescent probe during a freeze-thaw-sonication cycle in the presence of added liposomes containing cholesterol. In such a reconstituted system, glycine receptor agonists are able to increase the rate of anion influx into the vesicles. The action of agonists is blocked by the simultaneous presence of strychnine or other glycine antagonists. The results show that the purified 48,000- and 58,000-dalton polypeptides reconstituted into phospholipid vesicles can bind ligands and promote specific ion translocation in a way similar to the glycine receptor in its native environment.

  19. Comparative study on Charpy specimen reconstitution techniques

    International Nuclear Information System (INIS)

    Bourdiliau, B.; Decroix, G.-M.; Averty, X.; Wident, P.; Bienvenu, Y.

    2011-01-01

    Highlights: → Welding processes are used to reconstitute previously tested Charpy specimens. → Stud welding is preferred for a quick installation, almost immediately operational. → Friction welding produces better quality welds, but requires a development effort. - Abstract: Reconstitution techniques are often used to allow material from previously fractured Charpy-V specimens to be reused for additional experiments. This paper presents a comparative experimental study of various reconstitution techniques and evaluates the feasibility of these methods for future use in shielded cells. The following techniques were investigated: arc stud welding, 6.0 kW CO 2 continuous wave laser welding, 4.5 kW YAG continuous wave laser welding and friction welding. Subsize Charpy specimens were reconstituted using a 400 W YAG pulsed wave laser. The best result was obtained with arc stud welding; the resilience of the reconstituted specimens and the load-displacement curves agreed well with the reference specimens, and the temperature elevation caused by the welding process was limited to the vicinity of the weld. Good results were also obtained with friction welding; this process led to the best quality welds. Laser welding seems to have affected the central part of the specimens, thus leading to different resilience values and load-displacement curves.

  20. Common Developmental Tasks in Forming Reconstituted Families.

    Science.gov (United States)

    Kleinman, Judith

    1979-01-01

    Developmental tasks common to the formation of a reconstituted family are described, particularly the continued mourning of the old family; the formation of a solid marital relationship despite the difficulties presented by past failures and the presence of children; and the formation of sibling alliances across family lines. (Author)

  1. Serosal and Endometrial Reconstitution During Myomectomy ...

    African Journals Online (AJOL)

    Context: Myomectomy is usually performed when uterine fibroids are associated with infertility. Serosal and endometrial reconstitution are some of the subtle challenges that the gynaecologist has to deal with during myomectomy, in an attempt to minimise postoperative pelvic and intrauterine adhesions. Objective: This ...

  2. Detergent-Mediated Reconstitution of Membrane Proteins

    NARCIS (Netherlands)

    Knol, J; Sjollema, K.A; Poolman, B.

    1998-01-01

    The efficiency of reconstitution of the lactose transport protein (LacS) of Streptococcus thermophilus is markedly higher with Triton X-100 than with other detergents commonly employed to mediate the membrane insertion. To rationalize these differences, the lipid/detergent structures that are formed

  3. Immune Reconstitution Inflammatory Syndrome and Shingles ...

    African Journals Online (AJOL)

    Immune Reconstitution Inflammatory Syndrome and Shingles Associated with a Combined Paralysis of Three Oculomotor Nerves: A Case Report. ... CASE DETAILS: A 40 years old patient was seen for a pain of the right side of the face and a complete immobility of the eyeball. The diagnosis of V1 shingles with a pan uveitis ...

  4. Stability issues in reconstitution by weapon addition

    Energy Technology Data Exchange (ETDEWEB)

    Canavan, G.H.

    1997-08-01

    Reconstitution of strategic forces by the unilateral uploading of additional weapons from initially symmetric modest force levels reduces first strike stability. These changes are quantified and traced to changes in first and second strike costs in a model of missile exchanges in which both strikes are optimized analytically.

  5. Glucose transport machinery reconstituted in cell models.

    Science.gov (United States)

    Hansen, Jesper S; Elbing, Karin; Thompson, James R; Malmstadt, Noah; Lindkvist-Petersson, Karin

    2015-02-11

    Here we demonstrate the production of a functioning cell model by formation of giant vesicles reconstituted with the GLUT1 glucose transporter and a glucose oxidase and hydrogen peroxidase linked fluorescent reporter internally. Hence, a simplified artificial cell is formed that is able to take up glucose and process it.

  6. Reconstitution of Low Bandwidth Reaction History

    International Nuclear Information System (INIS)

    May, M.; Clancy, T.; Fittinghoff, D.; Gennaro, P.; Hagans, K.; Halvorson, G.; Lowry, M.; Perry, T.; Roberson, P.; Smith, D.; Teruya, A.; Blair, J.; Davis, B.; Hunt, E.; Emkeit, B.; Galbraith, J.; Kelly, B.; Montoya, R.; Nickel, G.; Ogle, J.; Wilson, K.; Wood, M.

    2004-01-01

    The goal of the Test Readiness Program is to transition to a 24 month test readiness posture and if approved move to an 18-month posture. One of the key components of the Test Readiness Program necessary to meet this goal is the reconstitution of the important diagnostics. Since the end of nuclear testing, the ability to field diagnostics on a nuclear test has deteriorated. Reconstitution of diagnostics before those who had experience in nuclear testing either retire or leave is essential to achieving a shorter test readiness posture. Also, the data recording systems have not been used since the end of testing. This report documents the reconstitution of one vital diagnostic: the low bandwidth reaction history diagnostic for FY04. Reaction history is one of the major diagnostics that has been used on all LLNL and LANL tests since the early days of nuclear testing. Reaction history refers to measuring the time history of the gamma and neutron output from a nuclear test. This gives direct information on the nuclear reactions taking place in the device. The reaction history measurements are one of the prime measurements the nuclear weapon scientists use to validate their models of device performance. All tests currently under consideration require the reaction history diagnostic. Thus moving to a shorter test readiness posture requires the reconstitution of the ability to make reaction history measurements. Reconstitution of reaction history was planned to be in two steps. Reaction history measurements that have been used in the past can be broadly placed into two categories. The most common type of reaction history and the one that has been performed on virtually all nuclear tests is termed low bandwidth reaction history. This measurement has a time response that is limited by the bandpass of kilometer length coaxial cables. When higher bandwidth has been required for specific measurements, fiber optic techniques have been used. This is referred to as high

  7. 7 CFR 58.522 - Reconstituting nonfat dry milk.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Reconstituting nonfat dry milk. 58.522 Section 58.522 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards... Procedures § 58.522 Reconstituting nonfat dry milk. Nonfat dry milk shall be reconstituted in a sanitary...

  8. 49 CFR 178.515 - Standards for reconstituted wood boxes.

    Science.gov (United States)

    2010-10-01

    ... wood boxes. (a) The identification code for a reconstituted wood box is 4F. (b) Construction requirements for reconstituted wood boxes are as follows: (1) The walls of boxes must be made of water... 49 Transportation 2 2010-10-01 2010-10-01 false Standards for reconstituted wood boxes. 178.515...

  9. The Psychodrama-Social Dramatics Separation.

    Science.gov (United States)

    Klepac, Richard L.

    Social dramatics is a therapeutic and educational program that can act as a mirror to reflect images of the self in action with others. It is the modality for experiential learning to correct social dysfunction by providing models for imitation, opportunities to practice and develop individual forms from that model, and risk free environments for…

  10. Puppetry and Creative Dramatics in Storytelling.

    Science.gov (United States)

    Champlin, Connie

    The 14 units in this book are designed to help teachers and librarians motivate children to express emotions, pantomime characters, and create a full story environment through puppetry and creative dramatics. Each unit is built around a specific piece of literature, and each provides instructions for the educator concerning the necessary…

  11. Reconstituted yogurt from yogurt cultured milk powder mix has better overall characteristics than reconstituted yogurt from commercial yogurt powder.

    Science.gov (United States)

    Song, Lijie; Aryana, Kayanush J

    2014-10-01

    For manufacture of commercial yogurt powder, yogurt has to go through a drying process, which substantially lowers the yogurt culture counts, so the potential health benefits of the yogurt culture bacteria are reduced. Also, upon reconstitution, commercial yogurt powder does not taste like yogurt and has an off-flavor. The objective was to study the microbial, physicochemical, and sensory characteristics of reconstituted yogurt from yogurt cultured milk powder (YCMP) mix and reconstituted yogurt from commercial yogurt powder (CYP). The CYP reconstituted yogurt was the control and YCMP mix reconstituted yogurt was the treatment. Microbial and physicochemical characteristics of the CYP reconstituted yogurt and YCMP mix reconstituted yogurt were analyzed daily for the first week and then weekly for a period of 8 wk. Sensory consumer testing of CYP reconstituted yogurt and YCMP mix reconstituted yogurt was conducted with 100 consumers. At 56 d, YCMP mix reconstituted yogurt had 5 log cfu/mL higher counts of Streptococcus thermophilus than the control (CYP reconstituted yogurt). Also, Lactobacillus bulgaricus counts of YCMP mix reconstituted yogurt were 6.55 log cfu/mL at 28 d and were 5.35 log cfu/mL at 56 d, whereas the CYP reconstituted yogurt from 28 d onwards had a count of yogurt also had significantly higher apparent viscosity and sensory scores for appearance, color, aroma, taste, thickness, overall liking, consumer acceptability, and purchase intent than CYP reconstituted yogurt. Overall, YCMP mix reconstituted yogurt had more desirable characteristics than CYP reconstituted yogurt. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  12. Reconstitution of active human core Mediator complex reveals a pivotal role of the MED14 subunit

    Science.gov (United States)

    Cevher, Murat A.; Shi, Yi; Li, Dan; Chait, Brian T.; Malik, Sohail; Roeder, Robert G.

    2014-01-01

    The evolutionarily conserved Mediator complex is a critical coactivator for RNA polymerase II (Pol II)-mediated transcription. Here, we report the reconstitution of a functional 15-subunit human core Mediator complex and its characterization by functional assays and chemical cross-linking coupled to mass spectrometry (CX-MS). Whereas the reconstituted head and middle modules can stably associate, only with incorporation of MED14 into the bi-modular complex does it acquire basal and coactivator functions. This results from a dramatically enhanced ability of MED14-containing complexes to associate with Pol II. Altogether, our analyses identify MED14 as both an architectural and a functional backbone of the Mediator complex. We further establish a conditional requirement for metazoan-specific MED26 that becomes evident in the presence of heterologous nuclear factors. This general approach paves the way for systematically dissecting the multiple layers of functionalities associated with the Mediator complex. PMID:25383669

  13. Reconstitution of active human core Mediator complex reveals a critical role of the MED14 subunit.

    Science.gov (United States)

    Cevher, Murat A; Shi, Yi; Li, Dan; Chait, Brian T; Malik, Sohail; Roeder, Robert G

    2014-12-01

    The evolutionarily conserved Mediator complex is a critical coactivator for RNA polymerase II (Pol II)-mediated transcription. Here we report the reconstitution of a functional 15-subunit human core Mediator complex and its characterization by functional assays and chemical cross-linking coupled to MS (CX-MS). Whereas the reconstituted head and middle modules can stably associate, basal and coactivator functions are acquired only after incorporation of MED14 into the bimodular complex. This results from a dramatically enhanced ability of MED14-containing complexes to associate with Pol II. Altogether, our analyses identify MED14 as both an architectural and a functional backbone of the Mediator complex. We further establish a conditional requirement for metazoan-specific MED26 that becomes evident in the presence of heterologous nuclear factors. This general approach paves the way for systematic dissection of the multiple layers of functionality associated with the Mediator complex.

  14. Care initiation area yields dramatic results.

    Science.gov (United States)

    2009-03-01

    The ED at Gaston Memorial Hospital in Gastonia, NC, has achieved dramatic results in key department metrics with a Care Initiation Area (CIA) and a physician in triage. Here's how the ED arrived at this winning solution: Leadership was trained in and implemented the Kaizen method, which eliminates redundant or inefficient process steps. Simulation software helped determine additional space needed by analyzing arrival patterns and other key data. After only two days of meetings, new ideas were implemented and tested.

  15. Reuse prevention syringes for reconstitution of lyophilized vaccines: Operational study and UNICEF plans for expanding introduction.

    Science.gov (United States)

    Fleming, Jessica A; Hoekstra, Edward John; Moniaga, Vanda; Widjaya, Anton; Soepardi, Jane; Supartha, Nyoman; Salovaara, Annika; Khamassi, Selma; Nelson, Carib

    2009-01-01

    Since the 1990s, the United Nation's Children's Fund has encouraged injection safety for immunizations through bundling vaccines with appropriate amounts of supporting equipment and by supplying autodisable (AD) syringes for injections. However, poor vaccine reconstitution practices continue to be reported worldwide. By 2009, UNICEF will begin to phase out the distribution of standard disposable syringes for vaccine reconstitution and replace them with reuse prevention (RUP) syringes, with a full transition expected by the end of 2010. A field evaluation in Indonesia was conducted to identify introduction requirements, issues with healthcare worker training and acceptance, and RUP syringe performance and safety. Managers and health workers felt that RUP syringes improved injection safety and fit easily into country logistical systems. Healthcare workers felt they were intuitive to use, but recommended special training. The integration of RUP reconstitution syringes by UNICEF could increase injection safety by preventing the reuse of syringes and reducing vaccine contamination.

  16. Cryptococcal Meningoencephalitis Relapse after an 8 Year Delay: An Interplay of Infection and Immune Reconstitution?

    Science.gov (United States)

    Katchanov, Juri; Blechschmidt, Cristiane; Nielsen, Kirsten; Branding, Gordian; Arastéh, Keikawus; Tintelnot, Kathrin; Meintjes, Graeme; Boulware, David R.; Stocker, Hartmut

    2016-01-01

    We report a case of a symptomatic relapse of HIV-related cryptococcal meningoencephalitis 8 years after the first diagnosis on the background of immune reconstitution. The findings as well as the clinical course suggests a combination of smouldering localized infection and enhanced inflammatory reaction related to immune restoration due to antiretroviral therapy. A combination of antifungal and anti-inflammatory therapy resulted in clinical and radiological improvement. Our case challenges the concept that immune reconstitution inflammatory syndrome and microbiological relapse are dichotomous entities. PMID:25505049

  17. Cryptococcal meningoencephalitis relapse after an eight-year delay: an interplay of infection and immune reconstitution.

    Science.gov (United States)

    Katchanov, Juri; Blechschmidt, Cristiane; Nielsen, Kirsten; Branding, Gordian; Arastéh, Keikawus; Tintelnot, Kathrin; Meintjes, Graeme; Boulware, David R; Stocker, Hartmut

    2015-10-01

    We report a case of a symptomatic relapse of HIV-related cryptococcal meningoencephalitis eight years after the first diagnosis on the background of immune reconstitution. The findings as well as the clinical course suggests a combination of smouldering localised infection and enhanced inflammatory reaction related to immune restoration due to antiretroviral therapy. A combination of antifungal and anti-inflammatory therapy resulted in clinical and radiological improvement. Our case challenges the concept that immune reconstitution inflammatory syndrome and microbiological relapse are dichotomous entities. © The Author(s) 2015.

  18. Immune Reconstitution Inflammatory Syndrome (IRIS): What pathologists should know.

    Science.gov (United States)

    Nelson, Ann Marie; Manabe, Yukari C; Lucas, Sebastian B

    2017-07-01

    Antiretroviral therapy has significantly improved the quality and length of life for those patients able to access effective and sustained treatment. The resulting restoration of the immune response is associated with a change in the clinical presentation of opportunistic infections, and the histologic reaction to pathogens. A complex combination of alterations in host response across the stages of HIV infection has been documented over the past 3 decades. The defects are seen in both acute and chronic phases of inflammation and involve innate and adaptive immunity. In advanced stages of HIV infection, the marked disruption of lymphoid tissue and loss of follicular dendritic cells limits the host's ability to process antigen and mount specific responses to pathogens. There are qualitative and quantitative defects in CD4 T cells due to HIV infection. The resulting indirect effects include loss of cytokine production, dysregulation of B-cell function, loss of cellular mediated immunity and "holes" in the immunologic repertoire that may not be restored with the use of antiretroviral therapy. Immune reconstitution allows the host to respond to and control infection, but a significant number of patients will have atypical inflammatory syndromes during the recovery period. We briefly discuss the impact of HIV infection on the immune system and give an overview of the spectrum of conditions attributed to the Immune Reconstitution Inflammatory syndrome (IRIS). Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Dramatization and attraction: new identification tools

    Directory of Open Access Journals (Sweden)

    V I Panteleeva

    2009-12-01

    Full Text Available The article provides the analysis of identification mechanisms and its presentation patterns in the contemporary megapolis, the reasons for identification challenges emerging in the contemporary multicultural world where the contemporary urban community offers the opportunity for identity transformation, so that the citizens of metropolis are faced with the challenge of finding new identification tools. The notions of «dramatization» and «social attraction» offered in the article for the identity problem analysis in the context of diverse style and taste options of the contemporary urban community serve to form an estimate of the behaviour of an individual who keeps deciding on his own behavioral pattern.

  20. Immune Reconstitution after Allogeneic Hematopoietic Stem Cell Transplantation

    Science.gov (United States)

    Ogonek, Justyna; Kralj Juric, Mateja; Ghimire, Sakhila; Varanasi, Pavankumar Reddy; Holler, Ernst; Greinix, Hildegard; Weissinger, Eva

    2016-01-01

    The timely reconstitution and regain of function of a donor-derived immune system is of utmost importance for the recovery and long-term survival of patients after allogeneic hematopoietic stem cell transplantation (HSCT). Of note, new developments such as umbilical cord blood or haploidentical grafts were associated with prolonged immunodeficiency due to delayed immune reconstitution, raising the need for better understanding and enhancing the process of immune reconstitution and finding strategies to further optimize these transplant procedures. Immune reconstitution post-HSCT occurs in several phases, innate immunity being the first to regain function. The slow T cell reconstitution is regarded as primarily responsible for deleterious infections with latent viruses or fungi, occurrence of graft-versus-host disease, and relapse. Here we aim to summarize the major steps of the adaptive immune reconstitution and will discuss the importance of immune balance in patients after HSCT. PMID:27909435

  1. An Analysis of Reconstituted Fluid Milk Pricing Policy

    OpenAIRE

    Glen D. Whipple

    1983-01-01

    This analysis suggests that alteration of the reconstituted fluid milk pricing provisions of federal and state milk market orders would have a substantial impact on market equilibrium. A reactive programming model of the U.S. milk market was used to simulate the effects of altered reconstituted fluid milk pricing policy. The solutions indicate that reconstituted fluid milk, as a lower cost alternative to fresh fluid milk, would make up a substantial portion of the fluid milk consumption in so...

  2. The dramatic extraction construction in French

    Directory of Open Access Journals (Sweden)

    Anne Abeillé

    2009-01-01

    Full Text Available Relying on spoken corpora (Corpaix, CRFP and on previous studies (Sabio 1995, 1996, we identify a construction common in spoken French, which we analyze as a particular case of extraction:a. dix sept ans il a. (Seventeen years he has [Corpaix]b. deux cigarettes j'ai fumé. (Two cigarettes I smoked [on the fly]The construction can only be a root clause and a declarative clause. Its interpretation is that of a thetic proposition. On the other hand, it is not associated with a unique information structure, since it is compatible with a focus-ground partition, with the extracted constotunet as a narrow focus, or with an all focus interpretation. We call this construction ‘dramatic extraction’, and the extracted element a ‘center’ (i.e. a focus or a figure. We formalize our analysis in the HPSG grammar.

  3. Assessment of anti-atherogenic drugs in vivo and reconstitution of lipoproteins using radioiodinated cholesteryl iopanoate

    International Nuclear Information System (INIS)

    DeGalan, M.R.

    1987-01-01

    A nonhydrolyzable radioiodinated cholesteryl ester, 125I-cholesteryl iopanoate (125I-Cl), was found to accumulate in high concentrations in atherosclerotic aortas of cholesterol-fed rabbits after intravenous administration. Aortas from normal chow-fed rabbits did not exhibit significant 125I-Cl accumulation. When cholesterol-fed rabbits were intravenously administered Tween-solubilized 125I-Cl and simultaneously treated with either of two anti-atherogenic compounds, estradiol 17β-cypionate or colestipol, the extent of aortic atherosclerosis was found to dramatically decrease. Measurement of aortic radioactivity was found to strongly correlate with the severity of atherosclerosis. Although the specificity of 125I-Cl for atheromatous lesions was very good, gamma-camera scintigraphy of the abdomens of these rabbits 6 days after cessation of 125I-Cl administration was not able to consistently predict the severity of atherosclerosis. Tissue distribution studies suggested that high blood and spinal column bone marrow radioactivity produced aorta:nontarget radioactivity ratios unfavorable with respect to imaging. To improve this ratio so as to permit noninvasive imaging, attempts were made to incorporate 125I-Cl into serum lipoproteins. Labelling of either rabbit LDL by in vivo incorporation or human LDL by transfer of 125I-Cl from liposomes using cholesteryl ester transfer protein resulted in lipoproteins with low specific activity. Higher specific activity was achieved by reconstituting delipidated human LDL with a mixture of 125I-Cl and unlabeled cholesteryl oleate. These particles were taken up in high amounts by monolayers of human fibroblasts but not by fibroblasts deficient in LDL receptors or by normal fibroblasts during competition with unlabeled native LDL

  4. Immune Reconstitution Inflammatory Syndrome in children

    Directory of Open Access Journals (Sweden)

    Nasreen Mahomed

    2017-11-01

    Full Text Available Immune Reconstitution Inflammatory Syndrome (IRIS refers to a collection of inflammatorydisorders, predominantly related to infectious processes that manifest after the initiation ofantiretroviral therapy (ART and can be classified as unmasking or paradoxical. The prevalenceof IRIS in children in sub-Saharan Africa is low. Approximately half of all cases are associatedwith Mycobacterium tuberculosis. It may be difficult to distinguish IRIS from tuberculosis andother opportunistic infections radiologically; therefore, radiological findings must be interpretedwith clinical and laboratory findings. In this review article, we describe the clinical andradiological manifestations of IRIS in children and provide illustrative radiological examples.

  5. NREL Discovers Enzyme Domains that Dramatically Improve Performance | News

    Science.gov (United States)

    of genomics data to find better enzymes, based on their genetic sequence alone. "In 10 years, it on these enzymes that can be targeted via genetic engineering to help break down cellulose faster to Decker, "At the time, tools for genetic engineering in Trichoderma were very limited, but we

  6. Thermostability of reconstituted newcastle disease virus strains at 36 ...

    African Journals Online (AJOL)

    Haemagglutination (HA) test was employed to determine the stability of HA titers of reconstituted form of Hitchner – B1 (B1), LaSota (L) and Komarov (K) strains of Newcastle Disease Vaccine (NDV) at 360c. The temperature treatment method was through incubation (in water bath) of the reconstituted vaccines at selected ...

  7. Stability of Beriplast P fibrin sealant: storage and reconstitution.

    Science.gov (United States)

    Eberhard, Ulrich; Broder, Martin; Witzke, Günther

    2006-04-26

    This study was performed to investigate the stability of Beriplast P fibrin sealant (FS) across a range of storage conditions, both pre- and post-reconstitution. Storage stability of the FS was evaluated during long-term refrigeration (24 months) with or without interim storage at elevated temperatures (40 degrees C for 1 week and 25 degrees C for 1 and 3 months). Stability of individual FS components was assessed by measuring: fibrinogen content, Factor XIII activity (FXIII), thrombin activity and aprotinin potency. The package integrity of each component was also checked (sterility testing, moisture content and pH). Storage stability was also evaluated by testing the reconstituted product for adhesion (tearing force testing after mixing the solutions) and sterility. Reconstitution stability was evaluated following 3-months' storage, for up to 50 h post-reconstitution using the same tests as for the storage stability investigations. Pre-defined specifications were met for fibrinogen content, Factor XIII activity, and thrombin activity, demonstrating storage stability. Package integrity and the functionality and sterility of the reconstituted product were confirmed throughout. Reconstitution stability was demonstrated for up to 50 h following reconstitution, in terms of both tearing force and sterility tests. In conclusion, the storage stability of Beriplast P was demonstrated over a range of 24-month storage schedules including interim exposure to elevated temperature, and the reconstituted product was stable for up to 50 h.

  8. Dramatic Outburst Reveals Nearest Black Hole

    Science.gov (United States)

    2000-01-01

    Scientists have discovered the closest black hole yet, a mere 1,600 light years from Earth. Its discovery was heralded by four of the most dramatic rapid X-ray intensity changes ever seen from one star. Astronomers from the Massachusetts Institute of Technology (MIT) and the National Science Foundation's National Radio Astronomy Observatory (NRAO) announced their findings at the American Astronomical Society's meeting in Atlanta. The black hole in the constellation Sagittarius, along with a normal star dubbed V4641 Sgr, form a violent system that briefly flooded part of our Milky Way Galaxy with X-rays and ejected subatomic particles moving at nearly the speed of light one day last September. At the peak of its X-ray output, V4641 Sgr was the brightest X-ray emitter in the sky. Astronomers call this type of system an X-ray nova because it suddenly becomes a bright source of X-rays, but this object shows characteristics never seen in an X-ray nova. "V4641 Sgr turns on and off so fast that it seems to represent a new subclass of X-ray novae," said Donald A. Smith, postdoctoral associate in MIT's Center for Space Research. Smith worked on data from this object with MIT principal research scientist Ronald Remillard and NRAO astronomer Robert Hjellming. "In X-rays, the intensity rose by a factor of more than 1,000 in seven hours, then dropped by a factor of 100 in two hours," Remillard said. The radio emission was seen as an image of an expanding "jet" of particles shooting out from the binary system. After reaching a maximum, the radio intensity dropped by a factor of nearly 40 within two days. "Radio telescopes give us a quick glimpse of something moving at a fantastically high velocity," Hjellming said. Black holes harbor enormous gravitational force that can literally rip the gas away from a nearby star. This transfer of gas is visible in many forms of radiation. Both orbiting X-ray telescopes and ground-based radio and optical telescopes saw the outburst of V4641

  9. Dramatic Advance in Quality in Flexographic Printing

    Directory of Open Access Journals (Sweden)

    Jochen Richter

    2004-12-01

    Full Text Available The enormous changes in flexography printing in recent years concerning the printing quality achievable cannot generally be ascribed to a single revolutionary invention, but are the result of continuous developments to the complete system. Thus the direct drive technology in all machine types and its associated advantages in terms of printing length corrections has become established since drupa 2000. The race for ever finer raster rolls has also been completed to the benefit of improvements in bowl geometry and in ceramic surfaces. Clearly improved colour transfer behaviour has become feasible as a result. In a closely intermeshed system such as flexography printing this naturally has to have an effect on the printing colours used. Further improvements in bonding agents and pigment concentrations now allow users to print ever thinner colour layers while maintaining all of the required authenticities.Furthermore, it has become possible to reduce additional disturbing characteristics in the UV colour area, such as the unpleasant odour. While the digital imaging of printing plates has primarily been improved in terms of economic efficiency by the use of up to eight parallel laser beams, extreme improvements in the system are noticeable especially in the area of directly engraved printing moulds. Whereas many still dismissed directly engraved polymer plates at the last drupa as a laboratory system, the first installation was recently placed on the market a mere three years later. A further noteworthy innovation of recent years that has reached market maturity is thin sleeve technology, which combines the advantages of a photopolymer plate with a round imaged printing mould. There are no high sleeve costs for each printing mould, except for one-off cost for an adapter sleeve. To conclude, it can be said that although flexography printing has experienced many new features in the time between drupa 2000 and today, it still has enormous potential for

  10. A dramatic, objective antiandrogen withdrawal response: case report and review of the literature

    Directory of Open Access Journals (Sweden)

    Litwin Alan

    2008-11-01

    Full Text Available Abstract Antiandrogen withdrawal response is an increasingly recognized entity in patients with metastatic prostate cancer. To our knowledge, there have been no reports describing a durable radiologic improvement along with prostate-specific antigen (PSA with discontinuation of the antiandrogen agent bicalutamide. We report a case in which a dramatic decline of serum PSA levels associated with a dramatic improvement in radiologic disease was achieved with bicalutamide discontinuation.

  11. The Challenges and Prospects of Teaching and Learning Dramatic ...

    African Journals Online (AJOL)

    Dramatic education is an all-inclusive academic discipline which uses as tools branches of learning that bear upon the dramatic impulse. It utilizes eclectically each and every single discipline into one unified body of knowledge, so that it can help man to comprehend the nature of experience. Educational drama or Dramatic ...

  12. Using dramatic role-play to develop emotional aptitude

    Directory of Open Access Journals (Sweden)

    Russell Dinapoli

    2009-12-01

    Full Text Available As university educators, we need to prepare students for the transition from the information age to what Daniel H. Pink (2005 calls the conceptual age, which is governed by artistry, empathy and emotion, by including in the curricula activities that stimulate both hemispheres of the brain. This can be done by promoting activities that energize what Daniel Goleman (1995 refers to as emotional intelligence, and it further maintains that, as Paul Ekman (2003 suggests, the ability to detect feelings improves communication. Recognizing the need to include in the curricula procedures that help develop students’ right brain aptitudes and enhance their communication skills, I have endeavoured to introduce dramatic scene study as a sustained activity in my English for Specific Purposes courses at the Universidad de Valencia. My aim was to energize the students’ creative and emotional aptitudes, as well as to dynamize effective teamwork. This article sustains that dramatic role-play, based on scripted scene study and related improvisational activities, is one way of achieving this.

  13. Functional reconstitution of an ATP-driven Ca2+-transport system from the plasma membrane of Commelina communis L

    International Nuclear Information System (INIS)

    Graef, P.; Weiler, E.W.

    1990-01-01

    The protein(s) that constitute(s) the ATP-driven Ca 2+ -translocator of plasma membrane enriched vesicles obtained by aqueous two-phase partitioning from leaves of Commelina communis L. has/have been solubilized and reincorporated into tightly sealed liposomes. The reconstituted Ca 2+ -transport system was studied using ATP-driven 45 Ca 2+ import into the proteoliposomes as a measure of activity. The detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonate proved to be the most suitable and was used at 10 millimolar concentration, i.e. just above its critical micellar concentration. The presence of additional phospholipid and ATP improved the solubilization and/or reconstitution. The characteristics of the reconstituted system were similar to those of the plasma membrane-bound activity, including the apparent K m for Ca 2+ inhibition by relatively high levels of vanadate and lacking response to added calmodulin. The reconstituted transport system was very strongly inhibited by erythrosine B and had a low apparent K m for ATP levels of the Ca 2+ -ionophore A 23187 instantaneously discharged 90% of the Ca 2+ associated with the vesicles, proving that it had been accumulated in the intravesicular volume in soluble, freely exchangeable form. Ca 2+ -transport in the reconstituted system was thus primary active, through a Ca 2+ -translocating ATPase

  14. effect of fines content on the engineering properties of reconstituted

    African Journals Online (AJOL)

    2012-11-03

    Nov 3, 2012 ... Reconstitution of the natural soil (0% fines content) yielded soil samples having fines content between ... action to different stabilizing agents are better under- .... tion cracking, whereas clay soils with too low plastic limit (PL) ...

  15. In vitro reconstitution of chaperone-mediated human RISC assembly.

    Science.gov (United States)

    Naruse, Ken; Matsuura-Suzuki, Eriko; Watanabe, Mariko; Iwasaki, Shintaro; Tomari, Yukihide

    2018-01-01

    To silence target mRNAs, small RNAs and Argonaute (Ago) proteins need to be assembled into RNA-induced silencing complexes (RISCs). Although the assembly of Drosophila melanogaster RISC was recently reconstituted by Ago2, the Dicer-2/R2D2 heterodimer, and five chaperone proteins, the absence of a reconstitution system for mammalian RISC assembly has posed analytical challenges. Here we describe reconstitution of human RISC assembly using Ago2 and five recombinant chaperone proteins: Hsp90β, Hsc70, Hop, Dnaja2, and p23. Our data show that ATP hydrolysis by both Hsp90β and Hsc70 is required for RISC assembly of small RNA duplexes but not for that of single-stranded RNAs. The reconstitution system lays the groundwork for further studies of small RNA-mediated gene silencing in mammals. © 2018 Naruse et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  16. Reconstitutable fuel assembly for a nuclear reactor

    International Nuclear Information System (INIS)

    Ferlan, S.J.; Kmonk, S.; Schallenberger, J.M.

    1982-01-01

    A reconstitutable fuel assembly for a nuclear reactor which includes a mechanical, rather than metallurgical, arrangement for connecting control rod guide thimbles to the top and bottom nozzles of a fuel assembly. Multiple sleeves enclosing control rod guide thimbles interconnect the top nozzle to the fuel assembly upper grid. Each sleeve is secured to the top nozzle by retaining rings disposed on opposite sides of the nozzle. Similar sleeves enclose the lower end of control rod guide thimbles and interconnect the bottom nozzle with the lowermost grid on the assembly. An end plug fitted in the bottom end of each sleeve extends through the bottom nozzle and is secured thereto by a retaining ring. Should it be necessary to remove a fuel rod from the assembly, the retaining rings in either the top or bottom nozzles may be removed to release the nozzle from the control rod guide thimbles and thus expose either the top or bottom ends of the fuel rods to fuel rod removing mechanisms

  17. Reconstituting botulinum toxin drugs: shaking, stirring or what?

    Science.gov (United States)

    Dressler, Dirk; Bigalke, Hans

    2016-05-01

    Most botulinum toxin (BT) drugs are stored as powders which need to be reconstituted with normal saline before clinical use. As botulinum neurotoxin (BNT), the therapeutically active ingredient, is a large double-stranded protein the process of reconstitution should be performed with special attention to mechanical stress applied. We wanted to test the mechanical stability of BNT during the reconstitution process. For this, 100 MU onabotulinumtoxinA (Botox(®), Irvine, CA, USA) was reconstituted with 2.0 ml of NaCl/H2O. Gentle reconstitution (GR) was performed with a 5 ml syringe, a 0.90 × 70 mm injection needle, one cycle of injection-aspiration-injection and two gentle shakes of the vial. Aggressive reconstitution (AR) was performed with a 5 ml syringe, a 0.40 × 40 mm injection needle, ten injection-aspiration-injection cycles and 30 s of continuous shaking of the vial. AR increased the time to paralysis in the mouse hemidiaphragm assay (HDA) from 72.0 ± 4.6 to 106.0 ± 16.0 min (*p = 0.002, two-tailed t test after Kolmogorov-Smirnova test with Lilliefors correction for normal distribution). Construction of a calibration curve revealed that the increase in the time to paralysis was correlated with a loss of potency of from 100 to 58 MU (-42 %). BT users should use large diameter injection needles for reconstitution, apply two or three injection-aspiration-injection cycles and, maybe, shake the vials a few times to rinse the entire glass wall. Aggressive reconstitution with small diameter needles, prolonged injection-aspiration-injection and violent shaking should be avoided.

  18. Concentration state dependence of the rheological and structural properties of reconstituted silk.

    Science.gov (United States)

    Mo, Chunli; Holland, Chris; Porter, David; Shao, Zhengzhong; Vollrath, Fritz

    2009-10-12

    The ability to control the processing of artificial silk is key to the successful application of this important and high performance biopolymer. Understanding where our current reconstitution process can be improved will not only aid us in the creation of better materials, but will also provide insight into the natural material along the way. This study aims to understand what proportion of reconstituted silk contributes to its rheological properties and what conformational state the silk proteins are in. It shows, for the first time, that a change in rheological properties can be related to a change in silk structures present in solution and reveals a low concentration gel state for silk that may have important implications for future successful artificial processing of silk.

  19. Immune reconstitution inflammatory syndrome after initiating highly active antiretroviral therapy in HIV-infected children

    International Nuclear Information System (INIS)

    Kilborn, Tracy; Zampoli, Marco

    2009-01-01

    The outcome of HIV infection has improved since the widespread availability of highly active antiretroviral therapy (HAART). Some patients, however, develop a clinical and radiological deterioration following initiation of HAART due to either the unmasking of occult subclinical infection or an enhanced inflammatory response to a treated infection. This phenomenon is believed to result from the restored ability to mount an immune response and is termed immune reconstitution inflammatory syndrome (IRIS) or immune reconstitution disease. IRIS is widely reported in the literature in adult patients, most commonly associated with mycobacterial infections. There is, however, a paucity of data documenting the radiological findings of IRIS in children. Radiologists need to be aware of this entity. As a diagnosis of exclusion it is essential that the radiological findings be assessed in the context of the clinical presentation. This article reviews the common clinical and radiological manifestations of IRIS in HIV-infected children. (orig.)

  20. Reconstitution of the activity of RND efflux pumps: a "bottom-up" approach.

    Science.gov (United States)

    Puvanendran, Dhenesh; Cece, Quentin; Picard, Martin

    2017-12-05

    Efflux pumps are systems devoted to the extrusion of noxious compounds. In this review, we discuss the various strategies that have thus far been undertaken for the investigation of efflux pumps after reconstitution into liposomes. It is challenging to uncover mechanisms and dynamics of efflux pumps due to a number of characteristics: their function depends on the correct assembly of three components and they span two adjacent membranes whose lipid compositions are very different. In addition, efflux pumps are active transporters that need energy to work. We present possible lines of improvement for the study of such systems and provide insights into future goals and challenges of efflux pump reconstitution and transport. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  1. Auditing Safety of Compounding and Reconstituting of Intravenous Medicines on Hospital Wards in Finland.

    Science.gov (United States)

    Suvikas-Peltonen, Eeva; Palmgren, Joni; Häggman, Verner; Celikkayalar, Ercan; Manninen, Raija; Airaksinen, Marja

    2017-01-01

    On the hospital wards in Finland, nurses generally reconstitute intravenous medicines, such as antibiotics, analgesics, and antiemetics prescribed by doctors. Medicine reconstitution is prone to many errors. Therefore, it is important to identify incorrect practices in the reconstitution of medicine to improve patient safety in hospitals. The aim of this study was to audit the compounding and reconstituting of intravenous medicines on hospital wards in a secondary-care hospital in Finland by using an assessment tool and microbiological testing for identifying issues posing patient safety risks. A hospital pharmacist conducted an external audit by using a validated 65-item assessment tool for safe-medicine compounding practices on 20 wards of the selected hospital. Also, three different microbiological samples were collected to assure the aseptics. Practices were evaluated using a four-point rating scale of "never performed," "rarely performed," "often performed," and "always performed," and were based on observation and interviews with nurses or ward pharmacists. In addition, glove-, settle plate-, and media fill-tests were collected. Associations between microbial sample results and audit-tool results were discussed. Altogether, only six out of the 65 items were fully implemented in all wards; these were related to logistic practices and quality assurance. More than half of the wards used incorrect practices ("rarely performed" or "never performed") for five items. Most of these obviated practices related to aseptic practices. All media-fill tests were clean but the number of colony forming units in glove samples and settle- plate samples varied from 0 to >100. More contamination was found in wards where environmental conditions were inadequate or the use of gloves was incorrect. Compounding practices were [mostly] quite well adapted, but the aseptic practices needed improvement. Attention should have been directed particularly to good aseptic techniques and

  2. Characterization and Purification of Polydisperse Reconstituted Lipoproteins and Nanolipoprotein Particles

    Directory of Open Access Journals (Sweden)

    Paul D. Hoeprich

    2009-07-01

    Full Text Available Heterogeneity is a fact that plagues the characterization and application of many self-assembled biological constructs. The importance of obtaining particle homogeneity in biological assemblies is a critical goal, as bulk analysis tools often require identical species for reliable interpretation of the results—indeed, important tools of analysis such as x-ray diffraction typically require over 90% purity for effectiveness. This issue bears particular importance in the case of lipoproteins. Lipid-binding proteins known as apolipoproteins can self assemble with liposomes to form reconstituted high density lipoproteins (rHDLs or nanolipoprotein particles (NLPs when used for biotechnology applications such as the solubilization of membrane proteins. Typically, the apolipoprotein and phospholipids reactants are self assembled and even with careful assembly protocols the product often contains heterogeneous particles. In fact, size polydispersity in rHDLs and NLPs published in the literature are frequently observed, which may confound the accurate use of analytical methods. In this article, we demonstrate a procedure for producing a pure, monodisperse NLP subpopulation from a polydisperse self-assembly using size exclusion chromatography (SEC coupled with high resolution particle imaging by atomic force microscopy (AFM. In addition, NLPs have been shown to self assemble both in the presence and absence of detergents such as cholate, yet the effects of cholate on NLP polydispersity and separation has not been systematically examined. Therefore, we examined the separation properties of NLPs assembled in both the absence and presence of cholate using SEC and native gel electrophoresis. From this analysis, NLPs prepared with and without cholate showed particles with well defined diameters spanning a similar size range. However, cholate was shown to have a dramatic affect on NLP separation by SEC and native gel electrophoresis. Furthermore, under

  3. Dramatics in the Classroom: Making Lessons Come Alive. Fastback 70.

    Science.gov (United States)

    Kelly, Elizabeth Flory

    The contents of this booklet focus on effective techniques for using dramatics in the classroom and are based on the premise that drama can integrate all skills and help to avoid fragmentation in learning. Chapters discuss motivating the dreamers in the classroom, curriculum dramatics--eclectic teaching, training the teacher, transforming the…

  4. Language in african dramatic literature: The example of Ola Rotimi's ...

    African Journals Online (AJOL)

    This paper discusses the use of language in African dramatic literature and identifies the controversy over the choice of a most appropriate language to be used by African playwrights and also X-rays the solution to language problem as proffered by prominent dramatic scholars. These solutions are then narrowed to Ola ...

  5. Exogenous endothelial cells as accelerators of hematopoietic reconstitution

    Directory of Open Access Journals (Sweden)

    Mizer J

    2012-11-01

    Full Text Available Abstract Despite the successes of recombinant hematopoietic-stimulatory factors at accelerating bone marrow reconstitution and shortening the neutropenic period post-transplantation, significant challenges remain such as cost, inability to reconstitute thrombocytic lineages, and lack of efficacy in conditions such as aplastic anemia. A possible means of accelerating hematopoietic reconstitution would be administration of cells capable of secreting hematopoietic growth factors. Advantages of this approach would include: a ability to regulate secretion of cytokines based on biological need; b long term, localized production of growth factors, alleviating need for systemic administration of factors that possess unintended adverse effects; and c potential to actively repair the hematopoietic stem cell niche. Here we overview the field of hematopoietic growth factors, discuss previous experiences with mesenchymal stem cells (MSC in accelerating hematopoiesis, and conclude by putting forth the rationale of utilizing exogenous endothelial cells as a novel cellular therapy for acceleration of hematopoietic recovery.

  6. Dramatically enhanced electrical breakdown strength in cellulose nanopaper

    Directory of Open Access Journals (Sweden)

    Jianwen Huang

    2016-09-01

    Full Text Available Electrical breakdown behaviors of nanopaper prepared from nanofibrillated cellulose (NFC were investigated. Compared to conventional insulating paper made from micro softwood fibers, nanopaper has a dramatically enhanced breakdown strength. Breakdown field of nanopaper is 67.7 kV/mm, whereas that of conventional paper is only 20 kV/mm. Air voids in the surface of conventional paper are observed by scanning electron microscope (SEM. Further analyses using mercury intrusion show that pore diameter of conventional paper is around 1.7 μm, while that of nanopaper is below 3 nm. Specific pore size of nanopaper is determined to be approximately 2.8 nm by the gas adsorption technique. In addition, theoretical breakdown strengths of nanopaper and conventional paper are also calculated to evaluate the effect of pore size. It turns out that theoretical values agree well with experimental data, indicating that the improved strength in nanopaper is mainly attributed to the decreased pore size. Due to its outstanding breakdown strength, this study indicates the suitability of nanopaper for electrical insulation in ultra-high voltage convert transformers and other electrical devices.

  7. Theodramatic Rehearsal: Fighting Self-Deception through the Dramatic Imagination

    Directory of Open Access Journals (Sweden)

    Brett Vaden

    2014-03-01

    Full Text Available This paper seeks to appropriate the insights of dramatic theology for Christian psychology and soul care. According to Kevin Vanhoozer, Scripture is the ‘script’ for human beings’ fitting participation in the acts and deeds of God in the world (i.e., ‘theodrama’. Keeping with this dramatic paradigm, the author will explore what ‘rehearsal’ might entail by drawing from a branch of psychotherapy called ‘psychodrama.’ The main question to be addressed in this appropriation of dramatic theology is, “How might dramatic rehearsal combat self-deception?” The author will only begin to answer this question, but in the attempt it is hoped that further reflection and clarity will be induced.

  8. Video Taping and Abnormal Psychology: Dramatized Clinical Interviews.

    Science.gov (United States)

    Lyons, Michael J.; And Others

    1984-01-01

    Students in an abnormal psychology course worked in teams to produce dramatizations of diagnostic interviews and then presented them in class. Positive and negative aspects of the activity are discussed. (RM)

  9. Grieg: Songs and dramatic works with orchestra. / Alan Blyth

    Index Scriptorium Estoniae

    Blyth, Alan

    1993-01-01

    Uuest heliplaadist "Grieg: Songs and dramatic works with orchestra. Barbara Bonney, Randi Stene, Hakan Hagegard, Ruth Tellefsen, Gothenburg Symphony Chorus and Orchestra, Neeme Järvi." CD 437 519 - 2GH

  10. Reusable locking tube in a reconstitutable fuel assembly

    International Nuclear Information System (INIS)

    Shallenberger, J.M.; Ferlan, S.J.

    1987-01-01

    This patent describes a reconstitutable fuel assembly including a top nozzle with an adapter plate having an interior wall forming at least one passageway, at least one guide thimble with an upper end portion, and an attaching structure having an outer socket formed by a circumferential groove defined in the adapter plate passageway wall and opening into the passageway and an inner socket formed by a circumferential bulge and at least one longitudinal slot defined in the upper end portion of the guide thimble. The circumferential bulge is capable of seating within the circumferential groove, an improved reusable tube for releasably locking the inner socket of the guide thimble upper end portion in locking engagement within the outer socket of the adapter plate passageway when the circumferential bulge is seated within the circumferential groove. The reusable tube comprises: (a) an elongated hollow tubular body capable of insertion within the adapter plate passageway and guide thimble upper end portion to a locking position therein such that the circumferential bulge of the inner socket is maintained seated in the locking engagement with the circumferential groove of the outer socket; and (b) at least a pair of dimples performed on the exterior of the tubular body prior to insertion of the body in the guide thimble upper end portion and to the locking position, the dimples being performed and configured to increase the thickness of the tubular body in relation to the remainder of the tubular body. The dimples are substantially resisting resilient yielding in relation to the remainder of the tubular body

  11. Reconstitution of Biosynthetic Machinery for the Synthesis of the Highly Elaborated Indole Diterpene Penitrem

    DEFF Research Database (Denmark)

    Liu, Chengwei; Tagami, Koichi; Minami, Atsushi

    2015-01-01

    KULNJ). Importantly, without conventional gene disruption, reconstitution of the biosynthetic machinery provided sufficient data to determine the pathway. It was thus demonstrated that the Aspergillus oryzae reconstitution system is a powerful method for studying the biosynthesis of complex natural products....

  12. Socio-Dramatic Affective-Relational Intervention for Adolescents with Asperger Syndrome & High Functioning Autism: Pilot Study

    Science.gov (United States)

    Lerner, Matthew D.; Mikami, Amori Yee; Levine, Karen

    2011-01-01

    This study examined the effectiveness of a novel intervention called "socio-dramatic affective-relational intervention" (SDARI), intended to improve social skills among adolescents with Asperger syndrome and high functioning autism diagnoses. SDARI adapts dramatic training activities to focus on in vivo practice of areas of social skill…

  13. Hanford Site waste tank farm facilities design reconstitution program plan

    International Nuclear Information System (INIS)

    Vollert, F.R.

    1994-01-01

    Throughout the commercial nuclear industry the lack of design reconstitution programs prior to the mid 1980's has resulted in inadequate documentation to support operating facilities configuration changes or safety evaluations. As a result, many utilities have completed or have ongoing design reconstitution programs and have discovered that without sufficient pre-planning their program can be potentially very expensive and may result in end-products inconsistent with the facility needs or expectations. A design reconstitution program plan is developed here for the Hanford waste tank farms facility as a consequence of the DOE Standard on operational configuration management. This design reconstitution plan provides for the recovery or regeneration of design requirements and basis, the compilation of Design Information Summaries, and a methodology to disposition items open for regeneration that were discovered during the development of Design Information Summaries. Implementation of this plan will culminate in an end-product of about 30 Design Information Summary documents. These documents will be developed to identify tank farms facility design requirements and design bases and thereby capture the technical baselines of the facility. This plan identifies the methodology necessary to systematically recover documents that are sources of design input information, and to evaluate and disposition open items or regeneration items discovered during the development of the Design Information Summaries or during the verification and validation processes. These development activities will be governed and implemented by three procedures and a guide that are to be developed as an outgrowth of this plan

  14. Family Therapy with Reconstituted Families: A Crisis-Induction Approach.

    Science.gov (United States)

    Baptiste, David A.

    1983-01-01

    Describes a crisis-based therapeutic approach for overcoming resistance in reconstituted families. Presents therapeutically induced crisis as a means through which therapists might purposefully disequilibrate families in which resistance is high and subsequently redirect them to meaningful change. Reviews implications and contraindications for the…

  15. Reconstitution of the fusogenic activity of vesicular stomatitis virus

    NARCIS (Netherlands)

    Metsikkö, K.; van Meer, G.; Simons, K.

    1986-01-01

    Enveloped virus glycoproteins exhibit membrane fusion activity. We have analysed whether the G protein of vesicular stomatitis virus, reconstituted into liposomes, is able to fuse nucleated cells in a pH-dependent fashion. Proteoliposomes produced by octylglucoside dialysis did not exhibit cell

  16. Chitosan Cross-linked Reconstituted Amniotic Collagen Membrane ...

    Indian Academy of Sciences (India)

    First page Back Continue Last page Overview Graphics. Chitosan Cross-linked Reconstituted Amniotic Collagen Membrane – An Excellent Cell Substratum. The KERATINOCYTE proliferation and Differentiation into multiple layers is due to the presence of type - IV collagen in the amnion. Cultured FIBROBLASTS had good ...

  17. Characterization of reconstituted partially purified glycerophosphate acyltransferase from Escherichia coli

    NARCIS (Netherlands)

    Kessels, J.M.M.; Bosch, H. van den

    1982-01-01

    A modification of the method of Snider and Kennedy (J. Bacteriol. (1977) 130, 1072–1083) was worked out to solubilize sn-glycero-3-phosphate acyltransferase from whole cells by Triton X-100. The solubilized preparation was used for a systematic study of the reconstitution of enzymatic activity as

  18. Reconstitution in yeast of the Arabidopsis SOS signaling pathway for Na+ homeostasis

    OpenAIRE

    Quintero, Francisco J.; Ohta, Masaru; Shi, Huazhong; Zhu, Jian-Kang; Pardo, José M.

    2002-01-01

    The Arabidopsis thaliana SOS1 protein is a putative Na H antiporter that functions in Na extrusion and is essential for the NaCl tolerance of plants. sos1 mutant plants share phenotypic similarities with mutants lacking the protein kinase SOS2 and the Ca2 sensor SOS3. To investigate whether the three SOS proteins function in the same response pathway, we have reconstituted the SOS system in yeast cells. Expression of SOS1 improved the Na tolerance of yeast mutants la...

  19. Reconstitution of apoglucose oxidase with FAD conjugates for biosensoring of progesterone

    NARCIS (Netherlands)

    Posthuma-Trumpie, G.A.; Berg, van den W.A.M.; Wiel, van de D.F.M.; Schaaper, W.M.M.; Korf, J.; Berkel, van W.J.H.

    2007-01-01

    The reconstitution of Aspergillus niger apoglucose oxidase (apoGOx) with FAD conjugates for biosensoring of progesterone was investigated. ApoGOx prepared by partial unfolding of the protein under acidic conditions consisted of reconstitutable monomers (50 ± 10%), reconstitutable dimers (20 ± 10%)

  20. The effect of lactase and formula reconstitution on milk osmolality.

    LENUS (Irish Health Repository)

    Malone, A J

    2012-02-03

    These experiments investigated the reaction rate of lactase on milk lactose by measuring milk osmolality; and explored the effect of formula reconstitution on milk osmolality. The investigations measured milk osmolality with the Fiske Os, freezing-point osmometer. Lactase (Lactaid) incubated with pure lactose solutions established the validity of the method. Lactase was incubated for 24 hours with four reconstituted milk formulas (Milumil, and Cow and Gate Nutrilon Plus, Farley\\'s First Milk, SMA Gold). Milk osmolality increased most rapidly in the first 4 hours after the addition of lactase. The lactase enzyme completed over 90% of the reaction within 12 hours. The milk osmolalities ranged from 487 to 591 mosm\\/kg after 24 hours with 2-4 drops of lactase in 240 ml of formula. A clinical guideline osmolality of 400 mosm\\/kg was reached in 240 ml of formula at 1 to 12 hours depending on the dose of lactase. High milk osmolalities due to prolonged enzyme incubation, or high lactase doses could be reduced to around 400 mosm\\/kg by dilution of 240 ml of formula with an extra 60 ml of water. The initial osmolality of formula after reconstitution by paediatric nurses varied widely and usually exceeded the manufacturer\\'s quoted osmolality. This initial osmolality was a further influence on the final osmolality reached after the addition of lactase. It is concluded that the recommended incubation time for Lactaid of 24 hours is unnecessary as lactase exerts the majority of its effect in less than 12 hours. Adjustment of Lactaid dose and incubation times will maintain milk formula osmolality within standard guidelines. Dilution with extra water will correct inadvertent high enzyme doses and prolonged incubation times. The normal method of reconstituting milk formulas from powder may be unreliable as the manufacturer\\'s quoted osmolality was not reproduced when milk formulas were reconstituted by paediatric nurses.

  1. Expression, purification, and reconstitution of the voltage-sensing domain from Ci-VSP.

    Science.gov (United States)

    Li, Qufei; Jogini, Vishwanath; Wanderling, Sherry; Cortes, D Marien; Perozo, Eduardo

    2012-10-16

    The voltage-sensing domain (VSD) is the common scaffold responsible for the functional behavior of voltage-gated ion channels, voltage sensitive enzymes, and proton channels. Because of the position of the voltage dependence of the available VSD structures, at present, they all represent the activated state of the sensor. Yet in the absence of a consensus resting state structure, the mechanistic details of voltage sensing remain controversial. The voltage dependence of the VSD from Ci-VSP (Ci-VSD) is dramatically right shifted, so that at 0 mV it presumably populates the putative resting state. Appropriate biochemical methods are an essential prerequisite for generating sufficient amounts of Ci-VSD protein for high-resolution structural studies. Here, we present a simple and robust protocol for the expression of eukaryotic Ci-VSD in Escherichia coli at milligram levels. The protein is pure, homogeneous, monodisperse, and well-folded after solubilization in Anzergent 3-14 at the analyzed concentration (~0.3 mg/mL). Ci-VSD can be reconstituted into liposomes of various compositions, and initial site-directed spin labeling and electron paramagnetic resonance (EPR) spectroscopic measurements indicate its first transmembrane segment folds into an α-helix, in agreement with the homologous region of other VSDs. On the basis of our results and enhanced relaxation EPR spectroscopy measurement, Ci-VSD reconstitutes essentially randomly in proteoliposomes, precluding straightforward application of transmembrane voltages in combination with spectroscopic methods. Nevertheless, these results represent an initial step that makes the resting state of a VSD accessible to a variety of biophysical and structural approaches, including X-ray crystallography, spectroscopic methods, and electrophysiology in lipid bilayers.

  2. Expression, Purification and Reconstitution of the Voltage Sensing Domain from Ci-VSP

    Science.gov (United States)

    Li, Qufei; Jogini, Vishwanath; Wanderling, Sherry; Cortes, D. Marien; Perozo, Eduardo

    2013-01-01

    The voltage-sensing domain (VSD) is the common scaffold responsible for the functional behavior of voltage gated ion channels, voltage sensitive enzymes and proton channels. Because of the position of the voltage dependence of the available VSD structures, at present, they all represent the activated state of the sensor. Yet, in the absence of a consensus resting state structure, the mechanistic details of voltage sensing remain controversial. The voltage dependence of the VSD from Ci-VSP (Ci-VSD) is dramatically right shifted, so that at 0 mV It presumably populates the putative resting state. Appropriate biochemical methods are an essential prerequisite to generate sufficient amounts of Ci-VSD protein for high-resolution structural studies. Here, we present a simple and robust protocol for the Escherichia coli expression of eukaryotic Ci-VSD at milligram levels. The protein is pure, homogeneous, mono-disperse and well folded after solubilization in Anzergent 3-14 at the analyzed concentration (~ 0.3 mg/mL). Ci-VSD can be reconstituted into liposomes of various compositions and initial site-directed spin labeling and EPR spectroscopic measurements indicate its first transmembrane segment folds into an α-helix, in agreement to the homologous region of other VSDs. Based on current results and enhanced relaxation EPR spectroscopy measurement, Ci-VSD reconstitutes essentially randomly in proteo-liposomes, precluding straightforward application of transmembrane voltages in combination with spectroscopic methods. Nevertheless, the present results represent an initial step that makes the resting state of a VSD accessible to a variety of biophysical and structural approaches, including X-ray crystallography, spectroscopic methods and electrophysiology in lipid bilayers. PMID:22989304

  3. How to Know when Dramatic Change Is on Track: Leading Indicators of School Turnarounds

    Science.gov (United States)

    Kowal, Julie; Ableidinger, Joe

    2011-01-01

    In recent years, national policymakers have placed new emphasis on "school turnarounds" as a strategy for rapid, dramatic improvement in chronically failing schools, calling on education leaders to turn around performance in the 5,000 lowest-achieving schools nationwide. This goal may seem daunting, given the dismal success rates of…

  4. Authoring for Engagement in Interactive Dramatic Experiences for Learning

    DEFF Research Database (Denmark)

    Schoenau-Fog, Henrik; Bruni, Luis Emilio; Khalil, Faysal Fuad

    2013-01-01

    ’s control over communicating a theme. To address this problem, this paper contributes a method for organizing narrative events in a free-roaming virtual environment. The Interactive Dramatic Experience Model (IDEM) retains the freedom of navigation while maintaining the possibility to construct various...

  5. On Dramatic Instruction: Towards a Taxonomy of Methods.

    Science.gov (United States)

    Courtney, Richard

    1987-01-01

    Examines the many possible methods used by instructors who work with dramatic action: in educational drama, drama therapy, social drama, and theater. Discusses an emergent taxonomy whereby instructors choose either spontaneous/formal, overt/covert/, or intrinsic/extrinsic methods. (JC)

  6. Poolside inspection, repair and reconstitution of LWR fuel elements

    International Nuclear Information System (INIS)

    1993-03-01

    The purpose of the meeting was to review the state of the art in the area of poolside inspection, repair and reconstitution of light water fuel elements. In the present publication it appears that techniques of inspection, repair and reconstitution of fuel elements have been developed by fuel suppliers and are now routinely and successfully applied in many countries. For the first time, the subject of control rod poolside examination was dealt with, poolside inspection and repair of a MOX assembly were reported and the inspection and repair of WWER assemblies were examined. Compared to the results of the previous meeting, present developments in the area aim now at reaching better economics, better reliability, reduction of personal doses and waste volume. Thirty-six participants representing twelve countries attended the meeting. Fifteen papers were presented in two sessions. An abstract was prepared for each of these papers. Refs, figs, tabs, diagrams, pictures and photos

  7. Reconstitution of the gastrointestinal microflora of lactobacillus-free mice.

    OpenAIRE

    Tannock, G W; Crichton, C; Welling, G W; Koopman, J P; Midtvedt, T

    1988-01-01

    A colony of mice that do not harbor lactobacilli in their digestive tracts but whose intestinal microflora is otherwise functionally similar to that of conventional animals was derived. Methods used to reconstitute the intestinal microflora of the mice included inoculation of the animals with cultures of specific microbes, noncultivable microbes attached to epithelial cells, and cecal contents from conventional mice treated with chloramphenicol. Twenty-six microflora-associated characteristic...

  8. Waste Encapsulation and Storage Facility (WESF) Design Reconstitution Plan

    International Nuclear Information System (INIS)

    HERNANDEZ, R.

    1999-01-01

    The purpose of Design Reconstitution is to establish a Design Baseline appropriate to the current facility mission. The scope of this plan is to ensure that Systems, Structures and Components (SSC) identified in the WESF Basis for Interim Operation (HNF-SDWM-BIO-002) are adequately described and documented, in order to support facility operations. In addition the plan addresses the adequacy of selected Design Topics which are also crucial for support of the facility Basis for Interim Operation (BIO)

  9. Chylous Ascites in a Patient with HIV/AIDS: A Late Complication of Mycobacterium avium Complex-Immune Reconstitution Inflammatory Syndrome

    Directory of Open Access Journals (Sweden)

    Imam H. Shaik

    2014-01-01

    Full Text Available Chylous ascites is very rare in HIV/AIDS and its association with Mycobacterium avium complex-immune reconstitution inflammatory syndrome (MAC-IRIS has been rarely reported. Here, we report a case of a young African-American male who developed chylous ascites as a late sequela to immune reconstitution inflammatory syndrome while on treatment for MAC. Antiretroviral drug-naive patients who start HAART in close proximity to the diagnosis of an opportunistic infection and have a rapid decline in HIV RNA level should be monitored for development of IRIS. Although the long term prognosis is poor, early diagnosis and treatment help to improve quality of life.

  10. Immunopathology of immune reconstitution inflammatory syndrome in Whipple's disease.

    Science.gov (United States)

    Moos, Verena; Feurle, Gerhard E; Schinnerling, Katina; Geelhaar, Anika; Friebel, Julian; Allers, Kristina; Moter, Annette; Kikhney, Judith; Loddenkemper, Christoph; Kühl, Anja A; Erben, Ulrike; Fenollar, Florence; Raoult, Didier; Schneider, Thomas

    2013-03-01

    During antimicrobial treatment of classic Whipple's disease (CWD), the chronic systemic infection with Tropheryma whipplei, immune reconstitution inflammatory syndrome (IRIS), is a serious complication. The aim of our study was to characterize the immunological processes underlying IRIS in CWD. Following the definition of IRIS, we describe histological features of IRIS and immunological parameters of 24 CWD IRIS patients, 189 CWD patients without IRIS, and 89 healthy individuals. T cell reconstitution, Th1 reactivity, and the phenotype of T cells were described in the peripheral blood, and infiltration of CD4(+) T cells and regulatory T cells in the duodenal mucosa was determined. During IRIS, tissues were heavily infiltrated by CD3(+), predominantly CD45RO(+)CD4(+) T cells. In the periphery, initial reduction of CD4(+) cell counts and their reconstitution on treatment was more pronounced in CWD patients with IRIS than in those without IRIS. The ratio of activated and regulatory CD4(+) T cells, nonspecific Th1 reactivity, and the proportion of naive among CD4(+) T cells was high, whereas serum IL-10 was low during IRIS. T. whipplei-specific Th1 reactivity remained suppressed before and after emergence of IRIS. The findings that IRIS in CWD mainly are mediated by nonspecific activation of CD4(+) T cells and that it is not sufficiently counterbalanced by regulatory T cells indicate that flare-up of pathogen-specific immunoreactivity is not instrumental in the pathogenesis of IRIS in CWD.

  11. Phenomenological and analytical interpretation of the rain-deposit relationship used for the reconstitution of the cesium 137 deposit in France consecutive to the accident of Chernobylsk

    International Nuclear Information System (INIS)

    Champion, D.; Renaud, Ph.; Metivier, J.M.; Garcia-Sanchez, L.; Pourcelot, L.

    2005-11-01

    The works presented in this report allow to give two conclusions: the first one bears on the reconstitution of the cesium 137 deposits induced by the Chernobylsk accident; the second one bears on the methods improvement of post accident consequences evaluation induced by the radioactive atmosphere fallout. (N.C.)

  12. T-cell independent reconstitution of the immunoglobulin levels in nu/nu mice

    International Nuclear Information System (INIS)

    Mannhardt, W.; Schulte-Wissermann, H.; Gardilcic, S.; Leon, F. de

    1982-01-01

    Nude mice were transplanted under the renal capsule either with allogeneic or human thymus that were long-term precultured or pretreated in vitro with Carrageenan for three days. None of the thymus tissue transplants showed lymphatic repopulation 9 wk after transplantation. Histological investigation of the peripheral lymphatic tissue did not reveal any change in the thymus-dependent area. On the other hand, plasma cells and germinal centers could be found in significantly increased numbers. In addition, a normalization of the serum immunoglobulin concentrations could be found, as no specific antibodies against thymus-dependent antigens were present after immunization and T-cell function did not improve. Similar results were obtained 9 wk after injection of irradiated thymocyte suspensions or of peritoneal macrophages from immunocompetent donors. It is concluded that thymus epithelial cells could act via macrophages on the polyclonal maturation and differentiation of B cells without involvement of T cells. This would be in agreement with the experience in some patients with severe combined immunodeficiency (SCID) in which reconstitution of the immunoglobulin levels is observed after transplantation of cultured thymus tissue before T-cell reconstitution can be demonstrated. (Auth.)

  13. Effect of hydrocolloids on the physico-chemical and rheological properties of reconstituted sweetened yoghurt powder.

    Science.gov (United States)

    Seth, Dibyakanta; Mishra, Hari Niwas; Deka, Sankar Chandra

    2018-03-01

    The consistency of sweetened yoghurt (misti dahi) is a desired characteristic which is attributed to the casein protein network formation during fermentation. Unfortunately, this property is lost in reconstituted sweetened yoghurt (RSY) due to the irreversible nature of protein denaturation during spray drying. Therefore, this study aimed to increase the consistency of RSY using different hydrocolloids. The effects addition of guar gum, pectin, κ-carrageenan and gelatin (0.1%w/v each) on the physico-chemical, microbial, rheological and sensory properties of RSY were investigated. RSY with 40% total solids demonstrated the rheological properties which are very similar to those of fresh sweetened yoghurt. RSY containing different hydrocolloids further increased the rheological properties. The dynamic rheological study revealed that the magnitude of storage modulus (G'), loss modulus (G″), and loss tangent (tan δ) were significantly influenced by the addition of hydrocolloids and gelatin exhibited highest dynamic moduli in RSY. However, κ-carrageenan added RSY was preferred sensorially as the rheological properties were very close to gelatin added RSY. Addition of hydrocolloids significantly increased the starter bacteria count and pH and reduced water expulsion rate (P < 0.05). Addition of hydrocolloids can improve the rheological properties of reconstituted yoghurt. The study concluded that the addition of κ-carrageenan showed better results in terms of rheological and sensory properties of RSY. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  14. Micro- and nanofabrication methods for ion channel reconstitution in bilayer lipid membranes

    Science.gov (United States)

    Tadaki, Daisuke; Yamaura, Daichi; Arata, Kohei; Ohori, Takeshi; Ma, Teng; Yamamoto, Hideaki; Niwano, Michio; Hirano-Iwata, Ayumi

    2018-03-01

    The self-assembled bilayer lipid membrane (BLM) forms the basic structure of the cell membrane and serves as a major barrier against ion movement. Ion channel proteins function as gated pores that permit ion permeation across the BLM. The reconstitution of ion channel proteins in artificially formed BLMs represents a well-defined system for investigating channel functions and screening drug effects on ion channels. In this review, we will discuss our recent microfabrication approaches to the formation of stable BLMs containing ion channel proteins as a potential platform for next-generation drug screening systems. BLMs formed in a microaperture having a tapered edge exhibited highly stable properties, such as a lifetime of ∼65 h and tolerance to solution changes even after the incorporation of the human ether-a-go-go-related gene (hERG) channel. We also explore a new method of efficiently incorporating human ion channels into BLMs by centrifugation. Our approaches to the formation of stable BLMs and efficient channel incorporation markedly improve the experimental efficiency of BLM reconstitution systems, leading to the realization of a BLM-based high-throughput platform for functional assays of various ion channels.

  15. Kazakhstan's Pension System: Pressures for Change and Dramatic Reforms

    OpenAIRE

    Seitenova, Ai-Gul S.; Becker, Charles M.

    2003-01-01

    Five years ago, Kazakhstan embarked on a dramatic reform of its pension and social security system in order to move from an unsustainable public defined benefit ("solidarity") system to one of defined mandatory contributions (accumulative system). While assessment of long-run success is premature, early results have exceeded expectations. This paper considers the reform's rationale and initial impact: Why did the Government of Kazakhstan decide to introduce a new pension system? What advantag...

  16. In vitro-reconstituted nucleoids can block mitochondrial DNA replication and transcription.

    Science.gov (United States)

    Farge, Géraldine; Mehmedovic, Majda; Baclayon, Marian; van den Wildenberg, Siet M J L; Roos, Wouter H; Gustafsson, Claes M; Wuite, Gijs J L; Falkenberg, Maria

    2014-07-10

    The mechanisms regulating the number of active copies of mtDNA are still unclear. A mammalian cell typically contains 1,000-10,000 copies of mtDNA, which are packaged into nucleoprotein complexes termed nucleoids. The main protein component of these structures is mitochondrial transcription factor A (TFAM). Here, we reconstitute nucleoid-like particles in vitro and demonstrate that small changes in TFAM levels dramatically impact the fraction of DNA molecules available for transcription and DNA replication. Compaction by TFAM is highly cooperative, and at physiological ratios of TFAM to DNA, there are large variations in compaction, from fully compacted nucleoids to naked DNA. In compacted nucleoids, TFAM forms stable protein filaments on DNA that block melting and prevent progression of the replication and transcription machineries. Based on our observations, we suggest that small variations in the TFAM-to-mtDNA ratio may be used to regulate mitochondrial gene transcription and DNA replication. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  17. Experimental study of low dose radiation stimulate the haematogenesis reconstitution of the recipient after bone marrow transplantation in mice

    International Nuclear Information System (INIS)

    Zhang Liyuan; Yang Shun; Zhang Ye; Zhang Mingzhi; Jiang Jiagui; Jiang Jianping

    2007-01-01

    Objective: To investigate if low dose radiation can stimulate the haematogenesis reconstitution of the recipient after bone marrow transplantation in mice. Methods: Bone marrow cells were irradiated in vitro by different low dose radiation and then cultured in vitro. 3 H-TdR incorporation was used to measure the reproductive activity of cells, and then the radiation dose with the best stimulating effect was determined. The donator myeloid cells were exposed to low dose radiation before the recipient mice received bone marrow transplantation; then the irradiated myeloid cells were infused to the recipient; and lastly, the counts of peripheral blood cells (PBC) and bone marrow mononuclear cells (BMMNC) were monitored in order to observe the effect of low dose radiation on haematogenesis reconstitution of the recipient animal after bone marrow transplantation. Results: The reproductive activity of the bone marrow cells irradiated by 6 and 8 cGy could be improved significantly in vitro. When the recipient mice received bone marrow transplantation of the myeloid cells after low dose radiation, the counts of BMMNC and PBC were higher than those in the control group (P<0.05). Conclusions: Low dose radiation can stimulate the haematogenesis reconstitution of the recipient after bone marrow transplantation. (authors)

  18. Reduced 99mTc labelled NCA-95/CEA-antibody uptake in liver due to gentle antibody reconstitution

    International Nuclear Information System (INIS)

    Reske, S.N.; Buell, U.

    1990-01-01

    The influence of reconstituting a murine monoclonal IgG 1 antibody kit with pertechnetate Tc99m on antibody distribution in the liver, spleen and sternal bone marrow of patients was examined. The 99m Tc-labelled antibody used is directed against non-specific cross-reacting antigen (NCA-95) and carcinoembryonic antigen (CEA) and has been successfully applied for imaging tissue inflammation and bone marrow scanning. Radioactivity uptake was determined in the liver, spleen, bone marrow and a precordial background region in a consecutive series of 25 patients, examined with an antibody preparation, routinely radiolabelled according to the manufacturer's recommendations and in 14 patients, in whom the antibody was reconstituted with special care, avoiding bubble formation and dropping of buffer into the antibody-containing vial. Gentle compared with routine antibody reconstitution caused a highly significant reduction of the antibody uptake in the liver, as determined by count densities, normalized to injected dose and acquisition time (13.2±5.5 vs 20.1±6.0 cpm per pixel, anti x±SD, P=0.008). The liver to background ratio was reduced from 3.4±1.4 to 1.9±0.5 (P<0.001). Spleen, sternal bone marrow and precordial background count rates were not significantly affected. These results clearly demonstrate that gentle antibody reconstitution can decrease non-specific antibody uptake in the liver by 34%±6.4% (anti x±SEM). Thus, scan quality is improved, and the potential deleterious camouflage of underlying structures is avoided. (orig.)

  19. Physical characterization of plakophilin 1 reconstituted with and without zinc.

    Science.gov (United States)

    Hofmann, I; Mücke, N; Reed, J; Herrmann, H; Langowski, J

    2000-07-01

    Plakophilin 1 (PKP1) belongs to the arm-repeat protein family which is characterized by the presence of a conserved 42-amino-acid motif. Despite individual members of the family containing a similar type of structural domain, they exhibit diverse cellular functions. PKP1 is ubiquitously expressed in human tissues and, depending on the type of cell, found prominently in the karyoplasm and/or in desmosomes. In surface plasmon resonance detection experiments, we noticed that PKP1 specifically bound zinc but not calcium or magnesium. Therefore we have used circular dichroism spectroscopy, limited proteolysis, analytical ultracentrifugation, electron microscopy and dynamic light scattering to establish the physical properties of recombinant PKP1 depending on the presence or absence of zinc. The alpha helix content of PKP1 was considerably higher when reconstituted with zinc than without. By atomic absorption spectroscopy 7.3 atoms zinc were shown to be tightly associated with one molecule of wild-type PKP1. The zinc-reconstituted protein formed globular particles of 21.9 +/- 8.4 nm diameter, as measured by electron microscopy after glycerol spraying/rotary metal shadowing. In parallel, the average sedimentation coefficient (s20, w) for zinc-containing PKP1 was 41S and its diffusion coefficient, as obtained by dynamic light scattering, 1.48 x 10-7 cm2.s-1. The molecular mass of 2.44 x 106 obtained from s and D yields an average stoichiometry of 30 for the PKP1 oligomer. In contrast, PKP1, reconstituted without zinc, contained no significant amount of zinc, sedimented with 4.6S, and was present in monomeric form as determined by sedimentation equilibrium centrifugation.

  20. Long term lymphocyte reconstitution after alemtuzumab treatment of multiple sclerosis

    KAUST Repository

    Hill-Cawthorne, Grant A.; Button, Tom; Tuohy, Orla C.; Jones, Joanne L.; May, Karen; Somerfield, Jennifer; Green, Alison J E; Giovannoni, Gavin; Compston, Alastair D.; Fahey, Michael T.; Coles, Alasdair J.

    2011-01-01

    Background: Alemtuzumab is a lymphocyte depleting monoclonal antibody that has demonstrated superior efficacy over interferon β-1a for relapsing-remitting multiple sclerosis (MS), and is currently under investigation in phase 3 trials. One unresolved issue is the duration and significance of the lymphopenia induced. The long term effects on lymphocyte reconstitution of a single course, and the consequences that this has on disability, morbidity, mortality and autoimmunity, were examined. Methods: The lymphocyte reconstitution (n=36; 384 person years) and crude safety data (n=37; 447 person years) are reported for the first patients with progressive MS to receive alemtuzumab (1991-1997). Reconstitution time was expressed as a geometric mean or, when a non-negligible number of individuals failed to recover, as a median using survival analysis. Results: Geometric mean recovery time (GMRT) of total lymphocyte counts to the lower limit of the normal range (LLN; ≥1.0×10 9 cells/l) was 12.7 months (95% CI 8.8 to 18.2 months). For B cells, GMRT to LLN (≥0.1×10 9/l) was 7.1 months (95% CI 5.3 to 9.5); median recovery times for CD8 (LLN ≥0.2×10 9 cells/l) and CD4 lymphocytes (LLN ≥0.4×10 9 cells/l) were 20 months and 35 months, respectively. However, CD8 and CD4 counts recovered to baseline levels in only 30% and 21% of patients, respectively. No infective safety concerns arose during 447 person years of follow-up. Conclusions: Lymphocyte counts recovered to LLN after a single course of alemtuzumab in approximately 8 months (B cells) and 3 years (T cell subsets), but usually did not recover to baseline values. However, this long lasting lymphopenia in patients with a previously normal immune system was not associated with an increased risk of serious opportunistic infection.

  1. Long term lymphocyte reconstitution after alemtuzumab treatment of multiple sclerosis

    KAUST Repository

    Hill-Cawthorne, Grant A.

    2011-11-05

    Background: Alemtuzumab is a lymphocyte depleting monoclonal antibody that has demonstrated superior efficacy over interferon β-1a for relapsing-remitting multiple sclerosis (MS), and is currently under investigation in phase 3 trials. One unresolved issue is the duration and significance of the lymphopenia induced. The long term effects on lymphocyte reconstitution of a single course, and the consequences that this has on disability, morbidity, mortality and autoimmunity, were examined. Methods: The lymphocyte reconstitution (n=36; 384 person years) and crude safety data (n=37; 447 person years) are reported for the first patients with progressive MS to receive alemtuzumab (1991-1997). Reconstitution time was expressed as a geometric mean or, when a non-negligible number of individuals failed to recover, as a median using survival analysis. Results: Geometric mean recovery time (GMRT) of total lymphocyte counts to the lower limit of the normal range (LLN; ≥1.0×10 9 cells/l) was 12.7 months (95% CI 8.8 to 18.2 months). For B cells, GMRT to LLN (≥0.1×10 9/l) was 7.1 months (95% CI 5.3 to 9.5); median recovery times for CD8 (LLN ≥0.2×10 9 cells/l) and CD4 lymphocytes (LLN ≥0.4×10 9 cells/l) were 20 months and 35 months, respectively. However, CD8 and CD4 counts recovered to baseline levels in only 30% and 21% of patients, respectively. No infective safety concerns arose during 447 person years of follow-up. Conclusions: Lymphocyte counts recovered to LLN after a single course of alemtuzumab in approximately 8 months (B cells) and 3 years (T cell subsets), but usually did not recover to baseline values. However, this long lasting lymphopenia in patients with a previously normal immune system was not associated with an increased risk of serious opportunistic infection.

  2. Dramatic response to levetiracetam in post-ischaemic Holmes’ tremor

    Science.gov (United States)

    Striano, P; Elefante, Andrea; Coppola, Antonietta; Tortora, Fabio; Zara, Federico; Minetti, Carlo

    2009-01-01

    Holmes’ tremor refers to an unusual combination of rest, postural and kinetic tremor of extremities. Common causes of Holmes’ tremor include stroke, trauma, vascular malformations and multiple sclerosis, with lesions involving the thalamus, brain stem or cerebellum. Although some drugs (eg, levodopa and dopaminergic drugs, clonazepam and propranolol) have been occasionally reported to give some benefit, medical treatment of Holmes’ tremor is unsatisfactory, and many patients require thalamic surgery to achieve satisfactory control. We report a patient in whom post-ischaemic Holmes’ tremor dramatically responded to levetiracetam treatment. PMID:21686707

  3. Confusion in the Study of Immune Reconstitution Inflammatory Syndrome

    Directory of Open Access Journals (Sweden)

    Claudia Alvarado-de la Barrera

    2017-05-01

    Full Text Available As a consequence of late presentation for HIV care, a significant proportion of individuals develop immune reconstitution inflammatory syndrome (IRIS soon after initiation of antiretroviral therapy. Incidence, predictors, and models of pathogenesis of IRIS vary in the literature. Here we discuss factors that may contribute to this lack of consensus. We propose that different pathogens drive different types of IRIS and suggest that these clinical conditions should be studied individually and not grouped under the general heading of “IRIS.”

  4. Underwater inspection, repair and reconstitution of water reactor fuel

    International Nuclear Information System (INIS)

    1988-06-01

    On-site fuel examination plays an important role for evaluation of fuel irradiation performance under reactor operating conditions. Also fuel assembly repairs are economically very attractive for operating nuclear utilities. The status of the processes of examination and repair, equipment used and research plans in Member States are reviewed in these Proceedings. All presentations were divided into three sessions: inspection diagnostic: facilities, techniques, and programmes (8 papers); repair and reconstitution techniques (6 papers); power plant experience (6 papers). A separate abstract was prepared for each of these 20 papers. Refs, figs and tabs

  5. An assessment of design control practices and design reconstitution programs in the nuclear power industry

    International Nuclear Information System (INIS)

    Imbro, E.V.

    1991-02-01

    The US Nuclear Regulatory Commission (NRC) and the utilities have identified shortcomings involving the maintenance of well-defined design bases and the availability of the necessary supporting design documentation. Many utilities have embarked on design-document reconstitution programs although there has been no clear consensus regarding what information should be included in design-bases documents, what is the minimum set of necessary design documents to support the design bases, or how missing or deficient design documentation should be handled. The NRC initiated a survey to ascertain the status of design control programs within the industry and the approaches to design-bases documentation used by some utilities. The survey scope included six utilities and one nuclear steam supply system vendor. Conclusions and observations resulting from the survey assessments are provided so that utilities and the NRC can consider actions to improve these programs. 12 refs

  6. Chronic Disseminated Candidiasis Complicated by Immune Reconstitution Inflammatory Syndrome in Child with Acute Lymphoblastic Leukemia

    Directory of Open Access Journals (Sweden)

    Olga Zając-Spychała

    2016-01-01

    Full Text Available Hepatosplenic candidiasis also known as chronic disseminated candidiasis is a rare manifestation of invasive fungal infection typically observed in patients with acute leukemia in prolonged, deep neutropenia. Immune reconstitution inflammatory syndrome (IRIS is an inflammatory disorder triggered by rapid resolution of neutropenia. Diagnosis and treatment of IRIS are still challenging due to a variety of clinical symptoms, lack of certain diagnostic criteria, and no standards of treatment. The diagnosis of IRIS is even more difficult in patients with hematological malignancies complicated by “probable” invasive fungal infection, when fungal pathogen is still uncertain. We report a case of probable hepatic candidiasis in 4-year-old boy with acute lymphoblastic leukemia. Despite proper antifungal therapy, there was no clinical and radiological improvement, so diagnosis of Candida-related IRIS was made and corticosteroid therapy was added to antifungal treatment achieving prompt resolution of infection symptoms.

  7. T cell reconstitution in allogeneic haematopoietic stem cell transplantation

    DEFF Research Database (Denmark)

    Kielsen, K; Jordan, K K; Uhlving, H H

    2015-01-01

    Infections and acute graft-versus-host disease (aGVHD) are major causes of treatment-related mortality and morbidity following allogeneic haematopoietic stem cell transplantation (HSCT). Both complications depend on reconstitution of the T-lymphocyte population based on donor T cells. Although...... it is well established that Interleukin-7 (IL-7) is a cytokine essential for de novo T cell development in the thymus and homoeostatic peripheral expansion of T cells, associations between circulating levels of IL-7 and T cell reconstitution following HSCT have not been investigated previously. We...... in patients treated with anti-thymocyte globulin (ATG) compared with those not treated with ATG (P = 0.0079). IL-7 levels at day +7 were negatively associated with T cell counts at day +30 to +60 (at day +60: CD3(+) : β = -10.6 × 10(6) cells/l, P = 0.0030; CD8(+) : β = -8.4 × 10(6) cells/l, P = 0.061; CD4...

  8. In vitro reconstitution of the active T. castaneum telomerase.

    Science.gov (United States)

    Schuller, Anthony P; Harkisheimer, Michael J; Skordalakes, Emmanuel

    2011-07-14

    Efforts to isolate the catalytic subunit of telomerase, TERT, in sufficient quantities for structural studies, have been met with limited success for more than a decade. Here, we present methods for the isolation of the recombinant Tribolium castaneum TERT (TcTERT) and the reconstitution of the active T. castaneum telomerase ribonucleoprotein (RNP) complex in vitro. Telomerase is a specialized reverse transcriptase that adds short DNA repeats, called telomeres, to the 3' end of linear chromosomes that serve to protect them from end-to-end fusion and degradation. Following DNA replication, a short segment is lost at the end of the chromosome and without telomerase, cells continue dividing until eventually reaching their Hayflick Limit. Additionally, telomerase is dormant in most somatic cells in adults, but is active in cancer cells where it promotes cell immortality. The minimal telomerase enzyme consists of two core components: the protein subunit (TERT), which comprises the catalytic subunit of the enzyme and an integral RNA component (TER), which contains the template TERT uses to synthesize telomeres. Prior to 2008, only structures for individual telomerase domains had been solved. A major breakthrough in this field came from the determination of the crystal structure of the active, catalytic subunit of T. castaneum telomerase, TcTERT. Here, we present methods for producing large quantities of the active, soluble TcTERT for structural and biochemical studies, and the reconstitution of the telomerase RNP complex in vitro for telomerase activity assays. An overview of the experimental methods used is shown in Figure 1.

  9. Information management needs for Fort Calhoun's design basis reconstitution project

    International Nuclear Information System (INIS)

    Beach, D.R.; Erickson, E.A.; Gambhir, S.K.; Parsons, R.D.

    1989-01-01

    While the need for information management is not new to the nuclear industry or Omaha Public Power District (OPPD), the interrelationship among design information, multiple systems, and design basis issues has necessitated the management of this information in new ways. The project team involved in the reconstitution of the design basis for OPPD's Fort Calhoun nuclear station has experienced the need for the developed effective methods for managing the vast amount of interrelated information associated with this effort. This management of information has been necessary to ensure that design basis documents (DBDs) adequately reflect the interrelated nature of component, system, and plant design; are complete and accurate; and are produced and maintained in a cost-effective manner. Fort Calhoun's aggressive design basis reconstitution project began in early 1987. The present scope of the project includes the production of 52 system and plant level DBDs; currently the project is ∼50% complete with DBDs in various stages of completion, from pilot DBDs through DBDs with approved formats, which have been issued for use. The experience in producing these documents has lead to a growing understanding of the special need for information management in each stage of the project. The development of the information tracking and management processes for the various stages of DBD development has proven to be cost-effective and gives a level of assurance that information has been included in the DBDs consistently and accurately

  10. Transmembrane topology of the acetylcholine receptor examined in reconstituted vesicles

    International Nuclear Information System (INIS)

    McCrea, P.D.

    1987-01-01

    Each of the five acetylcholine receptor (AChR) subunits, α 2 β-γδ, is believed to have the same number of transmembrane crossing and to share the same general folding pattern. AChR isolated from the electric organ of electric fish is predominantly dimeric. We have used this bridge as a marker for the C-terminus of the δ subunit, and presumably that of the other subunits in addition. The disulfide's accessibility to hydrophilic reductants, principally glutathione (GSH), was tested in a reconstituted vesicle system. The reduction of the δ-δ desulfide, as evidenced by the transition of AChrR dimers to monomers, was quantitatively monitored on velocity sedimentation sucrose gradients. Alternatively, the reduction of δ 2 to δ was followed by employing non-reducing SDS-PAGE. Reductants such as GSH were able to access the bridge in intact right-side-out vesicles. No acceleration of this process was evident when the vesicles were disrupted by freeze-thaw or by detergents. Control experiments which determined the rate of reduction of entrapped diphtheria toxin, or that of 3 H-GSH efflux, demonstrated that intact reconstituted vesicles provide an adequate permeability barrier to GSH access of their intravesicular space

  11. Phenomenological and analytical interpretation of the rain-deposit relationship used for the reconstitution of the cesium 137 deposit in France consecutive to the accident of Chernobylsk; Interpretation phenomenologique et analytique de la relation pluie-depot utilisee pour la reconstitution des depots de {sup 137}Cs en France consecutifs a l'accident de Tchernobyl

    Energy Technology Data Exchange (ETDEWEB)

    Champion, D.; Renaud, Ph.; Metivier, J.M.; Garcia-Sanchez, L.; Pourcelot, L. [Institut de Radioprotection et de Surete Nucleaire, Dir. de l' environnement et de l' intervention 92 - Fontenay-aux-Roses (France)

    2005-11-15

    The works presented in this report allow to give two conclusions: the first one bears on the reconstitution of the cesium 137 deposits induced by the Chernobylsk accident; the second one bears on the methods improvement of post accident consequences evaluation induced by the radioactive atmosphere fallout. (N.C.)

  12. Media and identification processes: a ritual and dramatic construction

    Directory of Open Access Journals (Sweden)

    Salomé Sola-Morales

    2016-07-01

    Full Text Available The present theoretical paper analyzes the way mediatic communication construct identifications. The thesis proposed is that the media configure ritually and dramatically identifications made by individuals and groups. First of all, we show the limitations of the concept of “identity” and we propose to use “identification” instead. Second, we explore the extent of social drama theory and role theory and we open it to new theoretical challenges and perspectives. Thirdly, it is shown that the concept of "ritual" is very useful to understand and interpret the identity construction through the media. All this leads us to conclude that both the theatricality and rituality are key to understanding the processes of identification that individuals and groups make in relation to the media.

  13. Dramatic response to nivolumab in xeroderma pigmentosum skin tumor.

    Science.gov (United States)

    Chambon, Fanny; Osdoit, Sophie; Bagny, Kelly; Moro, Anne; Nguyen, Jacqueline; Réguerre, Yves

    2018-02-01

    We report the case of a 6-year-old female with xeroderma pigmentosum (XP) who developed a nonoperable scalp tumor, treated with anti-programmed cell death protein 1 (anti-PD-1) therapy (nivolumab). She presented with a sarcomatoid carcinoma of the scalp with bone lysis as well as vascular and meningeal contact. Nivolumab was initiated because it has emerged as a promising immunotherapy. We observed a dramatic tumor response with excellent tolerance. However, while on nivolumab therapy she developed two large skin melanomas and several squamous cell carcinomas, which have been resected. These results demonstrate that cancer immunotherapy in patients with XP can be impressive but complex and warrants further investigation. © 2017 Wiley Periodicals, Inc.

  14. PROMOTING INCIDENTAL VOCABULARY LEARNING THROUGH VERBAL DRAMATIZATION OF WORDS

    Directory of Open Access Journals (Sweden)

    Looi-Chin Ch’ng

    2014-12-01

    Full Text Available Despite the fact that explicit teaching of vocabulary is often practised in English as a Second Language (ESL classrooms, it has been proven to be rather ineffective, largely because words are not taught in context. This has prompted the increasing use of incidental vocabulary learning approach, which emphasises on repeated readings as a source for vocabulary learning. By adopting this approach, this study aims to investigate students’ ability in learning vocabulary incidentally via verbal dramatization of written texts. In this case, readers’ theatre (RT is used as a way to allow learners to engage in active reading so as to promote vocabulary learning. A total of 160 diploma students participated in this case study and they were divided equally into two groups, namely classroom reading (CR and RT groups. A proficiency test was first conducted to determine their vocabulary levels. Based on the test results, a story was selected as the reading material in the two groups. The CR group read the story through a normal reading lesson in class while the RT group was required to verbally dramatize the text through readers’ theatre activity. Then, a post-test based on vocabulary levels was carried out and the results were compared. The findings revealed that incidental learning was more apparent in the RT group and their ability to learn words from the higher levels was noticeable through higher accuracy scores. Although not conclusive, this study has demonstrated the potential of using readers’ theatre as a form of incidental vocabulary learning activity in ESL settings.

  15. 27 CFR 25.263 - Production of concentrate and reconstitution of beer.

    Science.gov (United States)

    2010-04-01

    ... and reconstitution of beer. 25.263 Section 25.263 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS BEER Beer Concentrate § 25.263 Production of concentrate and reconstitution of beer. (a) Operations at brewery. A brewer may concentrate beer...

  16. Long-term reconstitution of dry barley increased phosphorus digestibility in pigs

    DEFF Research Database (Denmark)

    Ton Nu, Mai Anh; Blaabjerg, Karoline; Poulsen, Hanne Damgaard

    of reconstitution compared to dry stored barley on phosphorus (P) digestibility in pigs. Materials and Methods: Dry barley (13% moisture; phytate P, 1.7 g/kg DM) was rolled and stored directly or reconstituted with water to produce rolled barley with 35% moisture that was stored in air-tight conditions. After 49...

  17. Stark effect measurements on monomers and trimers of reconstituted light-harvesting complex II of plants

    NARCIS (Netherlands)

    Palacios, M.A.; Caffarri, S.; Bassi, R.; Grondelle, van R.; Amerongen, van H.

    2004-01-01

    The electric-field induced absorption changes (Stark effect) of reconstituted light-harvesting complex II (LHCII) in different oligomerisation states - monomers and trimers - with different xanthophyll content have been probed at 77 K. The Stark spectra of the reconstituted control samples,

  18. Mid-year report: IPC liaison and chemistry of thermal reconstitution

    International Nuclear Information System (INIS)

    Delegard, C.H.

    1995-05-01

    A program of investigation into the chemistry of alkaline Hanford Site tank waste is being conducted. The investigations have two main subtasks: liaison with the Institute of Physical Chemistry of the Russian Academy of Sciences and further laboratory testing of the chemistry of thermal reconstitution of Hanford Site tank waste. Progress to date includes: (1) a technical dialogue has been established with the Institute scientists; (2) editing of a technical literature review on the chemistry of the transuranic elements and technetium in alkaline media written by researchers at the Institute is complete; (3) four tasks from the Institute have been selected for support by the US Department of Energy; (4) technical information has been supplied to the Institute describing the composition of Hanford Site tank waste; (5) tests, using genuine waste from tank 104-S (a REDOX Process sludge), comparing the performance of thermal reconstitution with enhanced sludge washing show markedly improved dissolution of aluminum achieved by the thermal treatment; (6) a reduction/coprecipitation method was tested and shown to remove plutonium, solubilized by thermal treatment, rapidly and efficiently from solution; (7) technical chemistry support was provided to calciner kinetics tests at the University of Idaho; (8) tests to determine the speciation of plutonium and neptunium solubilized by thermal treatment show dissolved Pu(V) and Np(V) hydroxide complexes are produced, a Np(V) peroxide complex also was identified; (9) recently published data on Pu(IV) carbonate complexation in moderately alkaline (pH 12 to 13) solution led to reexamination of previous investigations of plutonium complexation in highly alkaline (3 to 5 molar NaOH) solutions

  19. Polyanhydride Nanoparticle Delivery Platform Dramatically Enhances Killing of Filarial Worms.

    Directory of Open Access Journals (Sweden)

    Andrea M Binnebose

    Full Text Available Filarial diseases represent a significant social and economic burden to over 120 million people worldwide and are caused by endoparasites that require the presence of symbiotic bacteria of the genus Wolbachia for fertility and viability of the host parasite. Targeting Wolbachia for elimination is a therapeutic approach that shows promise in the treatment of onchocerciasis and lymphatic filariasis. Here we demonstrate the use of a biodegradable polyanhydride nanoparticle-based platform for the co-delivery of the antibiotic doxycycline with the antiparasitic drug, ivermectin, to reduce microfilarial burden and rapidly kill adult worms. When doxycycline and ivermectin were co-delivered within polyanhydride nanoparticles, effective killing of adult female Brugia malayi filarial worms was achieved with approximately 4,000-fold reduction in the amount of drug used. Additionally the time to death of the macrofilaria was also significantly reduced (five-fold when the anti-filarial drug cocktail was delivered within polyanhydride nanoparticles. We hypothesize that the mechanism behind this dramatically enhanced killing of the macrofilaria is the ability of the polyanhydride nanoparticles to behave as a Trojan horse and penetrate the cuticle, bypassing excretory pumps of B. malayi, and effectively deliver drug directly to both the worm and Wolbachia at high enough microenvironmental concentrations to cause death. These provocative findings may have significant consequences for the reduction in the amount of drug and the length of treatment required for filarial infections in terms of patient compliance and reduced cost of treatment.

  20. Reconstituted AIM2 inflammasome in cell-free system.

    Science.gov (United States)

    Kaneko, Naoe; Ito, Yuki; Iwasaki, Tomoyuki; Takeda, Hiroyuki; Sawasaki, Tatsuya; Migita, Kiyoshi; Agematsu, Kazunaga; Kawakami, Atsushi; Morikawa, Shinnosuke; Mokuda, Sho; Kurata, Mie; Masumoto, Junya

    2015-11-01

    Absent in melanoma 2 (AIM2) is an intracellular pattern-recognition receptor, which is a member of the PYHIN protein family, consisting of a PYD domain and an IFN-inducible nuclear localization (HIN) domain. AIM2 is reported to oligomerize with adaptor protein ASC upon sensing bacterial and viral cytosolic DNA in order to form the AIM2 inflammasome, which activates caspase-1 leading to IL-1β secretion. Dysregulation of AIM2 inflammasome is supposed to result in autoinflammatory and autoimmune diseases. Thus, the development of new targeted drugs against AIM2 inflammasome would be important for the treatment of these diseases. However, since AIM2 inflammasome is an intracellular receptor, enforced internalization of both ligands and candidate molecules is necessary for the screening of AIM2-inflammasome-targeted molecules. We developed a reconstituted AIM2 inflammasome in a cell-free system with amplified luminescent proximity homogeneous assay (Alpha). Strong Alpha signal was detected upon incubation with poly-deoxyadenylic-deoxythymidylic acid, poly(dA:dT), whereas no Alpha signal was detected upon incubation with muramyl dipeptide, one of the NLR ligands of Nod2 ligand. The interaction between AIM2 and ASC was disrupted by an anti-human ASC monoclonal antibody, CRID3, a class of diarylsulfonylurea-containing compounds, and glycyrrhizin, a substance found in liquorice root. Thus, the reconstituted AIM2 inflammasome in a cell-free system is useful for screening AIM2-inflammasome-targeted therapeutic molecules. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. In vitro reconstitution of an abscisic acid signalling pathway

    KAUST Repository

    Fujii, Hiroaki; Chinnusamy, Viswanathan; Rodrigues, Americo; Rubio, Silvia; Antoni, Regina; Park, Sang-Youl; Cutler, Sean R.; Sheen, Jen; Rodriguez, Pedro L.; Zhu, Jian-Kang

    2009-01-01

    The phytohormone abscisic acid (ABA) regulates the expression of many genes in plants; it has critical functions in stress resistance and in growth and development. Several proteins have been reported to function as ABA receptors, and many more are known to be involved in ABA signalling. However, the identities of ABA receptors remain controversial and the mechanism of signalling from perception to downstream gene expression is unclear. Here we show that by combining the recently identified ABA receptor PYR1 with the type 2C protein phosphatase (PP2C) ABI1, the serine/threonine protein kinase SnRK2.6/OST1 and the transcription factor ABF2/AREB1, we can reconstitute ABA-triggered phosphorylation of the transcription factor in vitro. Introduction of these four components into plant protoplasts results in ABA-responsive gene expression. Protoplast and test-tube reconstitution assays were used to test the function of various members of the receptor, protein phosphatase and kinase families. Our results suggest that the default state of the SnRK2 kinases is an autophosphorylated, active state and that the SnRK2 kinases are kept inactive by the PP2Cs through physical interaction and dephosphorylation. We found that in the presence of ABA, the PYR/PYL (pyrabactin resistance 1/PYR1-like) receptor proteins can disrupt the interaction between the SnRK2s and PP2Cs, thus preventing the PP2C-mediated dephosphorylation of the SnRK2s and resulting in the activation of the SnRK2 kinases. Our results reveal new insights into ABA signalling mechanisms and define a minimal set of core components of a complete major ABA signalling pathway. © 2009 Macmillan Publishers Limited. All rights reserved.

  2. In vitro reconstitution of an abscisic acid signalling pathway

    KAUST Repository

    Fujii, Hiroaki

    2009-11-18

    The phytohormone abscisic acid (ABA) regulates the expression of many genes in plants; it has critical functions in stress resistance and in growth and development. Several proteins have been reported to function as ABA receptors, and many more are known to be involved in ABA signalling. However, the identities of ABA receptors remain controversial and the mechanism of signalling from perception to downstream gene expression is unclear. Here we show that by combining the recently identified ABA receptor PYR1 with the type 2C protein phosphatase (PP2C) ABI1, the serine/threonine protein kinase SnRK2.6/OST1 and the transcription factor ABF2/AREB1, we can reconstitute ABA-triggered phosphorylation of the transcription factor in vitro. Introduction of these four components into plant protoplasts results in ABA-responsive gene expression. Protoplast and test-tube reconstitution assays were used to test the function of various members of the receptor, protein phosphatase and kinase families. Our results suggest that the default state of the SnRK2 kinases is an autophosphorylated, active state and that the SnRK2 kinases are kept inactive by the PP2Cs through physical interaction and dephosphorylation. We found that in the presence of ABA, the PYR/PYL (pyrabactin resistance 1/PYR1-like) receptor proteins can disrupt the interaction between the SnRK2s and PP2Cs, thus preventing the PP2C-mediated dephosphorylation of the SnRK2s and resulting in the activation of the SnRK2 kinases. Our results reveal new insights into ABA signalling mechanisms and define a minimal set of core components of a complete major ABA signalling pathway. © 2009 Macmillan Publishers Limited. All rights reserved.

  3. Impacts on coralligenous outcrop biodiversity of a dramatic coastal storm.

    Directory of Open Access Journals (Sweden)

    Núria Teixidó

    Full Text Available Extreme events are rare, stochastic perturbations that can cause abrupt and dramatic ecological change within a short period of time relative to the lifespan of organisms. Studies over time provide exceptional opportunities to detect the effects of extreme climatic events and to measure their impacts by quantifying rates of change at population and community levels. In this study, we show how an extreme storm event affected the dynamics of benthic coralligenous outcrops in the NW Mediterranean Sea using data acquired before (2006-2008 and after the impact (2009-2010 at four different sites. Storms of comparable severity have been documented to occur occasionally within periods of 50 years in the Mediterranean Sea. We assessed the effects derived from the storm comparing changes in benthic community composition at sites exposed to and sheltered from this extreme event. The sites analyzed showed different damage from severe to negligible. The most exposed and impacted site experienced a major shift immediately after the storm, represented by changes in the species richness and beta diversity of benthic species. This site also showed higher compositional variability immediately after the storm and over the following year. The loss of cover of benthic species resulted between 22% and 58%. The damage across these species (e.g. calcareous algae, sponges, anthozoans, bryozoans, tunicates was uneven, and those with fragile forms were the most impacted, showing cover losses up to 50 to 100%. Interestingly, small patches survived after the storm and began to grow slightly during the following year. In contrast, sheltered sites showed no significant changes in all the studied parameters, indicating no variations due to the storm. This study provides new insights into the responses to large and rare extreme events of Mediterranean communities with low dynamics and long-lived species, which are among the most threatened by the effects of global change.

  4. The dramatic increase in social media in urology.

    Science.gov (United States)

    Matta, Rano; Doiron, Chris; Leveridge, Michael J

    2014-08-01

    Social media are established tools for rapid information dissemination to a broad audience. A major use has been the compilation of conference specific messaging known as tweets via preselected hashtags on Twitter. We analyzed Twitter use between consecutive years at the AUA (American Urological Association) and CUA (Canadian Urological Association) annual meetings. Publicly available tweets containing the established meeting hashtags were abstracted from an online archive. Tweets were categorized by author type and by content as informative (based on research presented at the conference) or uninformative (unrelated to presented data) according to an established classification scheme. We analyzed 5,402 tweets during the combined 18 meeting days, of which 4,098 were original and 1,304 were rebroadcast prior tweets. There was a large increase in Twitter use at the 2013 annual meetings compared to the 2012 meetings (4,591 tweets from a total of 540 accounts vs 811 from 134). Biotechnology analysts represented the highest volume of tweets (226 or 28%) in 2012 but in 2013 this majority shifted to urologists (2,765 or 60%). Of the tweets 29% were categorized as informative in 2012, which increased to 41% at the 2013 meetings. Twitter has emerged as a significant communication platform at urological meetings. Use increased dramatically between 2012 and 2013. Urologists have increasingly led this discussion with an increased focus on data arising from meeting proceedings. This adjunct to traditional meeting activity merits the attention of urologists and the professional associations that host such meetings. Copyright © 2014 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

  5. Stability of Reconstituted Telavancin Drug Product in Frozen Intravenous Bags.

    Science.gov (United States)

    Gu, Zhengtian; Wong, Anissa; Raquinio, Elvira; Nguyen, Alice

    2015-07-01

    Intravenous (IV) infusions of telavancin for injection are generally administered in-hospital, but in some circumstances they may be administered in an outpatient environment. In that setting, antibiotics may be premixed and frozen. This study determined the chemical stability of nonpreserved telavancin in various commonly used reconstitution diluents stored in IV bags (polyvinyl chloride [PVC] and PVC-free) at -20°C (-4°F) without light. Telavancin (750 mg/vial) was reconstituted with 5% dextrose injection USP (D5W) or 0.9% sodium chloride injection USP (NS) to obtain drug solutions at approximately 15 mg/mL. Infusion solutions of telavancin at diluted concentrations of 0.6 mg/mL and 8.0 mg/mL covering the range utilized in clinical practice were prepared in both PVC and PVC-free IV bags using D5W or NS solutions. The infusion solutions were stored under frozen conditions (-20°C ± 5°C [-4°F ± 41°F]) and the chemical stability was evaluated for up to 32 days. Telavancin concentration, purity, and degradant levels were determined using a stability-indicating high-performance liquid chromatography (HPLC) method. Telavancin IV infusion solutions in D5W or NS at 0.6 mg/mL and 8 mg/mL and stored at -20°C (-4°F) met the chemical stability criteria when tested on days 0, 7, 14, and 32. The assayed telavancin concentration at each time point was within 97% to 103% of the initial mean assay value. The total degradants quantified by the HPLC stability-indicating method did not show any significant change over the 32-day study period. Telavancin IV infusion solutions (in D5W or NS) in both PVC and PVC-free IV bags were stable for at least 32 days when stored at -20°C (-4°F) without light. These results provide prolonged frozen stability data further to that previously established for 7 days under refrigerated conditions (2°C-8°C [36°F -46°F]), and for 12 hours at room temperature when diluted into IV bags containing D5W, NS, or lactated Ringer's solution.

  6. An investigation into the dramatic increase in deaths from ...

    African Journals Online (AJOL)

    The first author collected the information on a cause of death form, a data form and an audit tool. As part of the quality-improvement project, 10 items were assessed to determine whether doctors adhered to treatment protocols, and 10 items were assessed to determine whether nurses adhered to nursing orders and basic ...

  7. PD-1 and PD-L1 inhibitors after platinum-based chemotherapy or in first-line therapy in cisplatin-ineligible patients: Dramatic improvement of prognosis and overall survival after decades of hopelessness in patients with metastatic urothelial cancer.

    Science.gov (United States)

    Resch, Irene; Shariat, Shahrokh F; Gust, Kilian M

    2018-01-01

    Until recently, there were no true innovations in the management of locally advanced (aUC) and metastatic urothelial cancer (mUC) in the last three decades. Vinflunine has been approved by the EMA (European Medicines Agency) with only limited improvement compared to best supportive care in second line treatment. In addition, gemcitabine/cisplatin has been established as an alternative to methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC). The advent of checkpoint inhibitors (CPI) revolutionized the care of these patients, transforming a unanimously deadly disease into one with hope through sustained disease control. Five immune CPI have recently been approved for aUC/mUC by the US Food and Drug Administration (FDA) including atezolizumab, nivolumab, pembrolizumab, durvalumab and avelumab. All five CPI are FDA-approved as second-line therapy with atezolizumab and pembrolizumab also being approved for first-line therapy in cisplatin-ineligible patients. The rapid acceptance in the treatment algorithm of UC is based on the impressive clinical efficacy of these agents in some patients, combined with their excellent safety profile. These new agents are indeed the most important advancement in UC care. However, the challenge in the age of precision medicine is to identify the patients who are most likely to benefit from CPIs, as the majority of patients do not respond to CPI. Toward this goal, validation of clinical, molecular and imaging biomarkers that serve for prediction and monitoring of treatment response are of central necessity.

  8. Comparison of immune reconstitution after allogeneic vs. autologous stem cell transplantation in 182 pediatric recipients

    Directory of Open Access Journals (Sweden)

    V. Wiegering

    2017-03-01

    Conclusion: Children undergoing a HSCT show a different pattern of immune reconstitution in the allogeneic and autologous setting. This might influence the outcome and should affect the clinical handling of infectious prophylaxis and re-vaccinations.

  9. Attempted reconstitution of a foal with primary severe combined immunodeficiency.

    Science.gov (United States)

    Campbell, T M; Studdert, M J; Ellis, W M; Paton, C M

    1983-07-01

    A foal with primary severe combined immunodeficiency, diagnosed within the first two weeks of life, was maintained with its dam in semi-isolation. The foal received continuous prophylactic antibiotic therapy, plasma from a sibling hyperimmunised with equine adenovirus vaccine, and intensive general nursing care. A full sibling female was selected as a bone marrow donor on the basis of red blood cell cross-matching and mixed lymphocyte reactions. Cyclophosphamide was given before two bone marrow transfusions at 35 and 73 days of age. To prevent graft versus host disease graft versus host disease the foal was maintained on methotrexate therapy. Reconstitution was not achieved nor were there signs of graft versus host disease. The foal died suddenly four days after the second bone marrow transfer when 77 days old. It had remained clinically free of any life threatening infectious disease and at necropsy a remarkable degree of freedom from infectious disease was confirmed. The most notable necropsy findings were bilateral nephrosis and myocardial degeneration and fibrosis. The likely cause of death was an electrolyte imbalance, particularly hypokalaemia, which secondarily affected the myocardium. Renal toxicity caused by the cytotoxic drugs, especially cyclophosphamide, may have contributed to the electrolyte imbalance.

  10. Reverse engineering GTPase programming languages with reconstituted signaling networks.

    Science.gov (United States)

    Coyle, Scott M

    2016-07-02

    The Ras superfamily GTPases represent one of the most prolific signaling currencies used in Eukaryotes. With these remarkable molecules, evolution has built GTPase networks that control diverse cellular processes such as growth, morphology, motility and trafficking. (1-4) Our knowledge of the individual players that underlie the function of these networks is deep; decades of biochemical and structural data has provided a mechanistic understanding of the molecules that turn GTPases ON and OFF, as well as how those GTPase states signal by controlling the assembly of downstream effectors. However, we know less about how these different activities work together as a system to specify complex dynamic signaling outcomes. Decoding this molecular "programming language" would help us understand how different species and cell types have used the same GTPase machinery in different ways to accomplish different tasks, and would also provide new insights as to how mutations to these networks can cause disease. We recently developed a bead-based microscopy assay to watch reconstituted H-Ras signaling systems at work under arbitrary configurations of regulators and effectors. (5) Here we highlight key observations and insights from this study and propose extensions to our method to further study this and other GTPase signaling systems.

  11. Heterologous Reconstitution of Omega-3 Polyunsaturated Fatty Acids in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Sun Hee Kim

    2015-01-01

    Full Text Available Reconstitution of nonnative, very-long-chain polyunsaturated fatty acid (VLC-PUFA biosynthetic pathways in Arabidopsis thaliana was undertaken. The introduction of three primary biosynthetic activities to cells requires the stable coexpression of multiple proteins within the same cell. Herein, we report that C22 VLC-PUFAs were synthesized from C18 precursors by reactions catalyzed by Δ6-desaturase, an ELOVL5-like enzyme involved in VLC-PUFA elongation, and Δ5-desaturase. Coexpression of the corresponding genes (McD6DES, AsELOVL5, and PtD5DES under the control of the seed-specific vicilin promoter resulted in production of docosapentaenoic acid (22:5 n-3 and docosatetraenoic acid (22:4 n-6 as well as eicosapentaenoic acid (20:5 n-3 and arachidonic acid (20:4 n-6 in Arabidopsis seeds. The contributions of the transgenic enzymes and endogenous fatty acid metabolism were determined. Specifically, the reasonable synthesis of omega-3 stearidonic acid (18:4 n-3 could be a useful tool to obtain a sustainable system for the production of omega-3 fatty acids in seeds of a transgenic T3 line 63-1. The results indicated that coexpression of the three proteins was stable. Therefore, this study suggests that metabolic engineering of oilseed crops to produce VLC-PUFAs is feasible.

  12. Reconstitution of Nucleosomes with Differentially Isotope-labeled Sister Histones.

    Science.gov (United States)

    Liokatis, Stamatios

    2017-03-26

    Asymmetrically modified nucleosomes contain the two copies of a histone (sister histones) decorated with distinct sets of Post-translational Modifications (PTMs). They are newly identified species with unknown means of establishment and functional implications. Current analytical methods are inadequate to detect the copy-specific occurrence of PTMs on the nucleosomal sister histones. This protocol presents a biochemical method for the in vitro reconstitution of nucleosomes containing differentially isotope-labeled sister histones. The generated complex can be also asymmetrically modified, after including a premodified histone pool during refolding of histone subcomplexes. These asymmetric nucleosome preparations can be readily reacted with histone-modifying enzymes to study modification cross-talk mechanisms imposed by the asymmetrically pre-incorporated PTM using nuclear magnetic resonance (NMR) spectroscopy. Particularly, the modification reactions in real-time can be mapped independently on the two sister histones by performing different types of NMR correlation experiments, tailored for the respective isotope type. This methodology provides the means to study crosstalk mechanisms that contribute to the formation and propagation of asymmetric PTM patterns on nucleosomal complexes.

  13. Active transport of Na+ by reconstituted Na,K-ATPase

    International Nuclear Information System (INIS)

    Boldyrev, A.A.; Svinukhova, I.A.

    1987-01-01

    The ability of ATP, CTP, ITP, GTP, and UTP to support ouabain-sensitive accumulation of Na + by proteoliposomes with a reconstituted Na/K-pump was investigated. At a low [Na + ]/[K + ] ratio in the medium (20 mM/50 mM), a correlation is observed between the proton-accepting capacity of the nucleotide and its effectiveness as a substrate of active transport. To test the hypothesis of the importance of the presence of a negative charge in the 1-position of the purine (3-pyrimidine) base of the nucleotide for mutual transitions between the Na- and K-conformations of Na,K-ATPase they used two analogs of ATP: N 1 -hydroxy-ATP, possessing proton acceptor capacity, and N 1 -methoxy-ATP, in the molecule of which the negative charge is quenched by a methyl group. The first substrate supports active accumulation of Na + in proteoliposomes at the same rate as ATP, whereas the second substrate is relatively ineffective

  14. Strength and Compressibility Characteristics of Reconstituted Organic Soil at Khulna Region of Bangladesh

    OpenAIRE

    Tahia Rabbee; Islam M. Rafizul

    2012-01-01

    This study depicts the experimental investigations into the effect of organic content on the shear strength and compressibility parameters of reconstituted soil. To these attempts, disturbed soil samples were collected from two selected locations of Khulna region. The reconstituted soil having organic content of 5-35 % were prepared in the laboratory to mix at various proportions of inorganic and organic soil at the water content equal to 1.25 times of liquid limits of collected samples .The ...

  15. Comparison of Shear Strength Properties for Undisturbed and Reconstituted Parit Nipah Peat, Johor

    Science.gov (United States)

    Azhar, A. T. S.; Norhaliza, W.; Ismail, B.; Abdullah, M. E.; Zakaria, M. N.

    2016-11-01

    Shear strength of soil is required to determine the soil stability and design the foundations. Peat is known as a soil with complex natural formations which also contributes problems to the researchers, developers, engineers and contractors in constructions and infrastructures. Most researchers conducted experiment and investigation of shear strength on peat using shear box test and simple shear test, but only a few had discovered the behavior of peat using triaxial consolidated undrained test. The aim of this paper is to determine the undrained shear strength properties of reconstituted peat and undisturbed peat of Parit Nipah, Johor for comparison purposes. All the reconstituted peat samples were formed with the size that passed opening sieve 3.35 mm and preconsolidation pressure at 100 kPa. The result of undrained shear strength of reconstituted peat was 21kPa for cohesion with the angle of friction, 41° compare to the undisturbed peat with cohesion 10 kPa and angle of friction, 16°. The undrained shear strength properties result obtained shows that the reconstituted peat has higher strength than undisturbed peat. For relationship deviator stress-strain, σd max and excess pore pressure, Δu, it shows that both of undisturbed and reconstituted gradually increased when σ’ increased, but at the end of the test, the values are slightly dropped. The physical properties of undisturbed and reconstituted peat were also investigated to correlate with the undrained shear strength results.

  16. World-Wide Effort Produces Dramatic "Movie" of Cosmic Jet

    Science.gov (United States)

    2001-05-01

    Astronomers using a world-wide collection of radio telescopes, including the National Science Foundation's Very Long Baseline Array (VLBA) of the National Radio Astronomy Observatory (NRAO), have made a dramatic "movie" of a voracious, superdense neutron star repeatedly spitting out subatomic particles at nearly the speed of light into two narrow jets as it pulls material from a companion star. The movie shows these jets ejecting clouds of hot plasma that are then "zapped" by pulses of energy in the jets as they move away from the neutron star. Frame from Radio-Telescope 'Movie' of Scorpius X-1 "We have directly measured the speed of energy flow in a cosmic jet for the first time," said Ed Fomalont, an astronomer at the NRAO in Charlottesville, Virginia. Fomalont worked with Barry Geldzahler and Charles Bradshaw of George Mason University in Fairfax, Virginia. The astronomers used the VLBA, the NSF's Very Large Array (VLA) and the Green Bank 140-foot telescope, along with radio telescopes from the European VLBI Network, Australia, Japan and South Africa to record the double-star system's eruptions continuously for 56 hours. "This study is going to be extremely valuable in helping us understand a phenomenon that we see throughout the universe," Fomalont said. Cosmic jets of superfast particles are ejected from the cores of numerous galaxies. On a smaller scale, similar jets are ejected from binary-star systems closer to home, in our own Milky Way Galaxy. While the jets from galaxy cores are thought to be powered by supermassive black holes millions of times more massive than the Sun, the closer "microquasars" are powered by much smaller black holes or by neutron stars only a few times more massive than the sun. "Studying one of the closer, smaller examples will help us understand how they all work, including the bigger ones," Geldzahler said. "The jets coming from distant galaxies are harder to study because of their much greater distance and the slowness of their

  17. Restricted Field IMRT Dramatically Enhances IMRT Planning for Mesothelioma

    International Nuclear Information System (INIS)

    Allen, Aaron M.; Schofield, Deborah; Hacker, Fred; Court, Laurence E.; Czerminska, Maria M.S.

    2007-01-01

    Purpose: To improve the target coverage and normal tissue sparing of intensity-modulated radiotherapy (IMRT) for mesothelioma after extrapleural pneumonectomy. Methods and Materials: Thirteen plans from patients previously treated with IMRT for mesothelioma were replanned using a restricted field technique. This technique was novel in two ways. It limited the entrance beams to 200 o around the target and three to four beams per case had their field apertures restricted down to the level of the heart or liver to further limit the contralateral lung dose. New constraints were added that included a mean lung dose of <9.5 Gy and volume receiving ≥5 Gy of <55%. Results: In all cases, the planning target volume coverage was excellent, with an average of 97% coverage of the planning target volume by the target dose. No change was seen in the target coverage with the new technique. The heart, kidneys, and esophagus were all kept under tolerance in all cases. The average mean lung dose, volume receiving ≥20 Gy, and volume receiving ≥5 Gy with the new technique was 6.6 Gy, 3.0%, and 50.8%, respectively, compared with 13.8 Gy, 15%, and 90% with the previous technique (p < 0.0001 for all three comparisons). The maximal value for any case in the cohort was 8.0 Gy, 7.3%, and 57.5% for the mean lung dose, volume receiving ≥20 Gy, and volume receiving ≥5 Gy, respectively. Conclusion: Restricted field IMRT provides an improved method to deliver IMRT to a complex target after extrapleural pneumonectomy. An upcoming Phase I trial will provide validation of these results

  18. Reconstitution of Biological Molecular generators of electric current. Bacteriorhodopsin.

    Science.gov (United States)

    Drachev, L A; Frolov, V N; Kaulen, A D; Liberman, E A; Ostroumov, S A; Plakunova, V G; Semenov, A Y; Skulachev, V P

    1976-11-25

    1. Photoinduced generation of electric current by bacteriorhodopsin, incorporated into the planar phospholipid membrane, has been directly measured with conventional electrometer techniques. 2. Two methods for bacteriorhodopsin incorporation have been developed: (a) formation of planar membrane from a mixture of decane solution of phospholipids and of the fraction of violet fragments of the Halobacterium halobium membrane (bacteriorhodopsin sheets), and (b) adhesion of bacteriorhodopsin-containing reconstituted spherical membranes (proteoliposomes) to the planar membrane in the presence of Ca2+ or some other cations. In both cases, illumination was found to induce electric current generation directed across the planar membrane, an effect which was measured by macroelectrodes immersed into electrolyte solutions on both sides of the membrane. 3. The maximal values of the transmembrane electric potential were of about 150 mV at a current of about 10(-11) A. The electromotive force measured by means of counterbalancing the photoeffect by an external battery, was found to reach the value of 300 mV. 4. The action spectrum of the photoeffect coincides with the bacteriorhodopsin absorption spectrum (maximum about 570 nm). 5. Both components of the electrochemical potential of H+ ions (electric potential and delta pH) across the planar membrane affect the bacteriorhodopsin photoelectric response in a fashion which could be expected if bacteriorhodopsin were a light-dependent electrogenic proton pump. 6. La3+ ions were shown to inhibit operation of those bacteriorhodopsin which pump out H+ ions from the La3+-containing compartment. 7. The photoeffect, mediated by proteoliposomes associated with thick planar membrane, is decreased by gramicidin A at concentrations which do not influence the planar membrane resistance in the light. On the contrary, a protonophorous uncoupler, trichlorocarbonylcyanidephenylhydrazone, decreases the photoeffect only if it is added at a

  19. Dramatically improved RNA in situ hybridization signals using LNA-modified probes

    DEFF Research Database (Denmark)

    Thomsen, Rune; Nielsen, Peter Stein; Jensen, Torben Heick

    2005-01-01

    . This increases the thermal stability of hybrids formed with RNA. The LNA-based probes detect specific RNAs in fixed yeast cells with an efficiency far better than conventional DNA oligonucleotide probes of the same sequence. Using this probe design, we were also able to detect poly(A)+ RNA accumulation within......In situ detection of RNA by hybridization with complementary probes is a powerful technique. Probe design is a critical parameter in successful target detection. We have evaluated the efficiency of fluorescent DNA oligonucleotides modified to contain locked nucleic acid (LNA) residues...

  20. Dramatic improvement of crystal quality for low-­temperature-grown rabbit muscle aldolase

    OpenAIRE

    Park, HaJeung; Rangarajan, Erumbi S.; Sygusch, Jurgen; Izard, Tina

    2010-01-01

    Rabbit muscle aldolase (RMA) was crystallized in complex with the low-complexity domain (LC4) of sorting nexin 9. Monoclinic crystals were obtained at room temperature that displayed large mosaicity and poor X-ray diffraction. However, orthorhombic RMA–LC4 crystals grown at 277 K under similar conditions exhibited low mosaicity, allowing data collection to 2.2 Å Bragg spacing and structure determination.

  1. Increasing magnetite contents of polymeric magnetic particles dramatically improves labeling of neural stem cell transplant populations.

    Science.gov (United States)

    Adams, Christopher F; Rai, Ahmad; Sneddon, Gregor; Yiu, Humphrey H P; Polyak, Boris; Chari, Divya M

    2015-01-01

    Safe and efficient delivery of therapeutic cells to sites of injury/disease in the central nervous system is a key goal for the translation of clinical cell transplantation therapies. Recently, 'magnetic cell localization strategies' have emerged as a promising and safe approach for targeted delivery of magnetic particle (MP) labeled stem cells to pathology sites. For neuroregenerative applications, this approach is limited by the lack of available neurocompatible MPs, and low cell labeling achieved in neural stem/precursor populations. We demonstrate that high magnetite content, self-sedimenting polymeric MPs [unfunctionalized poly(lactic acid) coated, without a transfecting component] achieve efficient labeling (≥90%) of primary neural stem cells (NSCs)-a 'hard-to-label' transplant population of major clinical relevance. Our protocols showed high safety with respect to key stem cell regenerative parameters. Critically, labeled cells were effectively localized in an in vitro flow system by magnetic force highlighting the translational potential of the methods used. Copyright © 2015 Elsevier Inc. All rights reserved.

  2. Central nervous system immune reconstitution inflammatory syndrome in AIDS: experience of a Mexican neurological centre.

    Science.gov (United States)

    Guevara-Silva, Erik A; Ramírez-Crescencio, María A; Soto-Hernández, José Luís; Cárdenas, Graciela

    2012-09-01

    Highly active antiretroviral therapy (HAART) restores the inflammatory immune response in AIDS patients and it may unmask previous subclinical infections or paradoxically exacerbate symptoms of opportunistic infections. Up to 25% of patients receiving HAART develop immune reconstitution inflammatory syndrome (IRIS). We describe six patients with IRIS central nervous system (CNSIRIS) manifestations emphasizing the relevance of CSF cultures and neuroimaging in early diagnosis and management. Patients with CNSIRIS were identified among hospitalized HIV-infected patients that started HAART from January 2002 through December 2007 at a referral neurological center in Mexico. One-hundred and forty-two HIV-infected patients with neurological signs were hospitalized, 64 of which had received HAART, and six (9.3%) developed CNSIRIS. Five patients were male. Two cases of tuberculosis, two of cryptococcosis, one of brain toxoplasmosis, and one possible PML case were found. IRIS onset occurred within 12 weeks of HAART in five patients. Anti-infective therapy was continued. In one case, HAART was temporarily suspended. In long-term follow-up the clinical condition improved in all patients. CNSIRIS associated to opportunistic infections appeared in 9% of patients receiving HAART. Interestingly, no cases of malignancy or neoplasm IRIS-related were found. Frequent clinical assessment and neuroimaging studies supported diagnosis and treatment. Risk factors were similar to those found in other series. Copyright © 2012 Elsevier B.V. All rights reserved.

  3. Reconstitutable nuclear reactor fuel assembly with unitary removable top nozzle subassembly

    International Nuclear Information System (INIS)

    Shallenberger, J.M.

    1987-01-01

    A reconstitutable fuel assembly is described having at least one control rod guide thimble and a top nozzle, the guide thimble including an upper extension, the top nozzle including at least one hold-down spring, an upper hold-down plate and a lower adapter plate, an improved attaching structure removably mounting the top nozzle as a unitary subassembly on the guide thimble. The attaching structure comprises: (a) a coupling member interfitting the lower adapter plate, the upper hold-down plate and the hold-down spring disposed between the plates so as to capture and retain the plates and spring together as a unitary subassembly in which the upper plate is slidably moveable along the coupling member relative to the lower plate with the spring biasing the upper plate away from the lower plate. The coupling member has spaced apart upper and lower portions with a central passageway extending for slidably receiving the upper extension of the guide thimble in a nonattached relationship in which the coupling member is slidably movable relative to the guide thimble extension for respectively inserting and removing the coupling member on and from the guide thimble extension

  4. Thermal-Stability and Reconstitution Ability of Listeria Phages P100 and A511

    Directory of Open Access Journals (Sweden)

    Hanie Ahmadi

    2017-12-01

    Full Text Available The study evaluated the thermal-stability of Listeria phages P100 and A511 at temperatures simulating the preparation of ready-to-eat meats. The phage infectivity after heating to 71°C and holding for a minimum of 30 s, before eventually cooling to 4°C were examined. Higher temperatures of 75, 80, and 85°C were also tested to evaluate their effect on phages thermal-stability. This study found that despite minor differences in the amino acid sequences of their structural proteins, the two phages responded differently to high temperatures. P100 activity declined at least 10 log (PFU mL-1 with exposure to 71°C (30 s and falling below the limit of detection (1 log PFU mL-1 while, A511 dropped from 108 to 105 PFU mL-1. Cooling resulted in partial reconstitution of P100 phage particles to 103 PFU mL-1. Exposure to 75°C (30 s abolished A511 activity (8 log PFU mL-1 and both phages showed reconstitution during cooling phase after exposure to 75°C. P100 exhibited reconstitution after treatment at 80°C (30 s, conversely A511 showed no reconstitution activity. Heating P100 to 85°C abolished the reconstitution potential. Substantial differences were found in thermal-stability and reconstitution of the examined phages showing A511 to be more thermo-stable than P100, while P100 exhibited reconstitution during cooling after treatment at 80°C which was absent in A511. The differences in predicted melting temperatures of structural proteins of P100 and A511 were consistent with the observed differences in thermal stability and morphological changes observed with transmission electron microscopy.

  5. Development of reconstitution technique of irradiated specimens. 3. Report for FY 1995 and FY 1996 on JAERI-IHI cooperated research program (joint research)

    Energy Technology Data Exchange (ETDEWEB)

    Nishiyama, Yutaka; Fukaya, Kiyoshi; Onizawa, Kunio; Suzuki, Masahide [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment; Nakamura, Terumi; Kaihara, Shoichiro; Yoshida, Kazuo; Sato, Akira

    1998-10-01

    The cooperated research between Japan Atomic Energy Research Institute and Ishikawajima-Harima Heavy Industries Co., Ltd. on the development of reconstitution technique of irradiated reactor pressure vessel surveillance specimens has been performed from FY 1993. In FY 1993-1994, the method of surface activated joining (SAJ) was applied to reconstitution of Charpy impact specimens. Some verification tests using unirradiated reactor pressure vessel plate materials have shown that SAJ is feasible for a reconstitution technique, in particular, owing to low joining temperature. The present paper reports the results of the cooperated research performed in FY 1995-1996. To improve the quality of the SAJ, the configuration of the end tab surface to be joined with the insert material was modified. The torque measured during joining was also introduced in joining parameters. A nondestructive inspection, temperature measurements in the specimens during joining were performed. The effect of joining on Charpy impact properties was discussed. For practical application of the technique to irradiated specimens, we confirmed that the impact specimens with joining interface gave rise to no failure at the joining position during impact test after neutron irradiation. (author)

  6. Production of Conjugated Linoleic and Conjugated α-Linolenic Acid in a Reconstituted Skim Milk-Based Medium by Bifidobacterial Strains Isolated from Human Breast Milk

    Directory of Open Access Journals (Sweden)

    María Antonia Villar-Tajadura

    2014-01-01

    Full Text Available Eight bifidobacterial strains isolated from human breast milk have been tested for their abilities to convert linoleic acid (LA and α-linolenic acid (LNA to conjugated linoleic acid (CLA and conjugated α-linolenic acid (CLNA, respectively. These bioactive lipids display important properties that may contribute to the maintenance and improvement human health. Three selected Bifidobacterium breve strains produced CLA from LA and CLNA from LNA in MRS (160–170 and 210–230 μg mL−1, resp. and, also, in reconstituted skim milk (75–95 and 210–244 μg mL−1, resp.. These bifidobacterial strains were also able to simultaneously produce both CLA (90–105 μg mL−1 and CLNA (290–320 μg mL−1 in reconstituted skim milk. Globally, our findings suggest that these bifidobacterial strains are potential candidates for the design of new fermented dairy products naturally containing very high concentrations of these bioactive lipids. To our knowledge, this is the first study describing CLNA production and coproduction of CLA and CLNA by Bifidobacterium breve strains isolated from human milk in reconstituted skim milk.

  7. Socio-dramatic affective-relational intervention for adolescents with asperger syndrome & high functioning autism: pilot study.

    Science.gov (United States)

    Lerner, Matthew D; Mikami, Amori Yee; Levine, Karen

    2011-01-01

    This study examined the effectiveness of a novel intervention called 'socio-dramatic affective-relational intervention' (SDARI), intended to improve social skills among adolescents with Asperger syndrome and high functioning autism diagnoses. SDARI adapts dramatic training activities to focus on in vivo practice of areas of social skill deficit among this population. SDARI was administered as a six-week summer program in a community human service agency. Nine SDARI participants and eight age- and diagnosis-group matched adolescents not receiving SDARI were compared on child- and parent-report of social functioning at three week intervals beginning six weeks prior to intervention and ending six weeks post-intervention. Hierarchical Linear Modeling (HLM) was used to estimate growth trends between groups to assess treatment outcomes and post-treatment maintenance. Results indicated significant improvement and post-treatment maintenance among SDARI participants on several measures of child social functioning. Implications for practice and research are discussed.

  8. SLAM family markers are conserved among hematopoietic stem cells from old and reconstituted mice and markedly increase their purity.

    Science.gov (United States)

    Yilmaz, Omer H; Kiel, Mark J; Morrison, Sean J

    2006-02-01

    Recent advances have increased the purity of hematopoietic stem cells (HSCs) isolated from young mouse bone marrow. However, little attention has been paid to the purity of HSCs from other contexts. Although Thy-1 low Sca-1+ Lineage- c-kit+ cells from young bone marrow are highly enriched for HSCs (1 in 5 cells gives long-term multilineage reconstitution after transplantation into irradiated mice), the same population from old, reconstituted, or cytokine-mobilized mice engrafts much less efficiently (1 in 78 to 1 in 185 cells gives long-term multilineage reconstitution). To test whether we could increase the purity of HSCs isolated from these contexts, we examined the SLAM family markers CD150 and CD48. All detectable HSCs from old, reconstituted, and cyclophosphamide/G-CSF-mobilized mice were CD150+ CD48-, just as in normal young bone marrow. Thy-1 low Sca-1+ Lineage- c-kit+ cells from old, reconstituted, or mobilized mice included mainly CD48+ and/or CD150- cells that lacked reconstituting ability. CD150+ CD48- Sca-1+ Lineage- c-kit+ cells from old, reconstituted, or mobilized mice were much more highly enriched for HSCs, with 1 in 3 to 1 in 7 cells giving long-term multilineage reconstitution. SLAM family receptor expression is conserved among HSCs from diverse contexts, and HSCs from old, reconstituted, and mobilized mice engraft relatively efficiently after transplantation when contaminating cells are eliminated.

  9. Reconstitution of a Kv channel into lipid membranes for structural and functional studies.

    Science.gov (United States)

    Lee, Sungsoo; Zheng, Hui; Shi, Liang; Jiang, Qiu-Xing

    2013-07-13

    To study the lipid-protein interaction in a reductionistic fashion, it is necessary to incorporate the membrane proteins into membranes of well-defined lipid composition. We are studying the lipid-dependent gating effects in a prototype voltage-gated potassium (Kv) channel, and have worked out detailed procedures to reconstitute the channels into different membrane systems. Our reconstitution procedures take consideration of both detergent-induced fusion of vesicles and the fusion of protein/detergent micelles with the lipid/detergent mixed micelles as well as the importance of reaching an equilibrium distribution of lipids among the protein/detergent/lipid and the detergent/lipid mixed micelles. Our data suggested that the insertion of the channels in the lipid vesicles is relatively random in orientations, and the reconstitution efficiency is so high that no detectable protein aggregates were seen in fractionation experiments. We have utilized the reconstituted channels to determine the conformational states of the channels in different lipids, record electrical activities of a small number of channels incorporated in planar lipid bilayers, screen for conformation-specific ligands from a phage-displayed peptide library, and support the growth of 2D crystals of the channels in membranes. The reconstitution procedures described here may be adapted for studying other membrane proteins in lipid bilayers, especially for the investigation of the lipid effects on the eukaryotic voltage-gated ion channels.

  10. Solubilization and reconstitution of the formylmethionylleucylphenylalanine receptor coupled to guanine nucleotide regulatory protein

    International Nuclear Information System (INIS)

    Williamson, K.; Dickey, B.F.; Pyun, H.Y.; Navarro, J.

    1988-01-01

    The authors describe the solubilization, resolution, and reconstitution of the formylmethionylleucylphenylalanine (fMet-Leu-Phe) receptor and guanine nucleotide regulatory proteins (G-proteins). The receptor was solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate. Guanine nucleotides decreased the number of high-affinity binding sites and accelerated the rate of dissociation of the receptor-ligand complex, suggesting that the solubilized receptor remained coupled to endogenous G-proteins. The solubilized receptor was resolved from endogenous G-proteins by fractionation on a wheat germ agglutinin (WGA)-Sepharose 4B column. High-affinity [ 3 H]fMet-Leu-Phe binding to the WGA-purified receptor was diminished and exhibited reduced guanine nucleotide sensitivity. High-affinity [ 3 H]fMET-Leu-Phe binding and guanine nucleotide sensitivity were reconstituted upon the addition of purified brain G-proteins. Similar results were obtained when the receptor was reconstituted with brain G-proteins into phospholipid vesicles by gel filtration chromatography. In addition, they also demonstrated fMET-Leu-Phe-dependent GTP hydrolysis in the reconstituted vesicles. The results of this work indicate that coupling of the fMet-Leu-Phe receptor to G-proteins converts the receptor to a high-affinity binding state and that agonist produces activation of G-proteins. The resolution and functional reconstitution of this receptor should provide an important step toward the elucidation of the molecular mechanism of the fMet-Leu-Phe transduction system in neutrophils

  11. Dramatization at Extracurricular Activities as a Means to Enhance Foreign Language Teaching

    Directory of Open Access Journals (Sweden)

    Marina Valeryevna Kuimova

    2015-02-01

    Full Text Available The main purpose of foreign language teaching is to learn to communicate and overcome the language barrier. The article studies dramatization and its appropriateness in foreign language teaching, provides criteria for choosing a literary work for dramatization. The use of dramatization at extracurricular activities develops communication abilities, creativity; enhances motivation to learn a foreign language and strengthens students’ confidence in a foreign language.

  12. Surface reconstitution of glucose oxidase onto a norbornylogous bridge self-assembled monolayer

    International Nuclear Information System (INIS)

    Liu Jingquan; Paddon-Row, Michael N.; Gooding, J. Justin

    2006-01-01

    An electrode construct was fabricated in which a self-assembled monolayer containing a novel norbornylogous bridge was covalently attached to flavin adenine dinucleotide (FAD), the redox active centre of several oxidase enzymes. The electrochemistry of the construct was investigated before and after the reconstitution of glucose oxidase around the surface bound FAD. Rapid rates of electron transfer were observed both before and after the reconstitution of biocatalytically active enzyme. However, no biocatalytic activity was observed under anaerobic conditions suggesting the a lack of enzyme turnover through direct electron transfer. It is proposed that a decrease in the electronic coupling between the redox active FAD and the electrode following reconstitution of the glucose oxidase - a probable consequence of the FAD being immersed in a protein environment - was responsible for the inability of the enzyme to be turned over under anaerobic conditions

  13. Nanodisc-Tm: Rapid functional assessment of nanodisc reconstituted membrane proteins by CPM assay.

    Science.gov (United States)

    Ashok, Yashwanth; Jaakola, Veli-Pekka

    2016-01-01

    Membrane proteins are generally unstable in detergents. Therefore, biochemical and biophysical studies of membrane proteins in lipidic environments provides a near native-like environment suitable for membrane proteins. However, manipulation of proteins embedded in lipid bilayer has remained difficult. Methods such as nanodiscs and lipid cubic phase have been developed for easy manipulation of membrane proteins and have yielded significant insights into membrane proteins. Traditionally functional reconstitution of receptors in nanodiscs has been studied with radioligands. We present a simple and faster method for studying the functionality of reconstituted membrane proteins for routine characterization of protein batches after initial optimization of suitable conditions using radioligands. The benefits of the method are •Faster and generic method to assess functional reconstitution of membrane proteins.•Adaptable in high throughput format (≥96 well format).•Stability measurement in near-native lipid environment and lipid dependent melting temperatures.

  14. Subunit Organisation of In Vitro Reconstituted HOPS and CORVET Multisubunit Membrane Tethering Complexes

    Science.gov (United States)

    Guo, Zhong; Johnston, Wayne; Kovtun, Oleksiy; Mureev, Sergey; Bröcker, Cornelia; Ungermann, Christian; Alexandrov, Kirill

    2013-01-01

    Biochemical and structural analysis of macromolecular protein assemblies remains challenging due to technical difficulties in recombinant expression, engineering and reconstitution of multisubunit complexes. Here we use a recently developed cell-free protein expression system based on the protozoan Leishmania tarentolae to produce in vitro all six subunits of the 600 kDa HOPS and CORVET membrane tethering complexes. We demonstrate that both subcomplexes and the entire HOPS complex can be reconstituted in vitro resulting in a comprehensive subunit interaction map. To our knowledge this is the largest eukaryotic protein complex in vitro reconstituted to date. Using the truncation and interaction analysis, we demonstrate that the complex is assembled through short hydrophobic sequences located in the C-terminus of the individual Vps subunits. Based on this data we propose a model of the HOPS and CORVET complex assembly that reconciles the available biochemical and structural data. PMID:24312556

  15. Visualization and quantitative analysis of reconstituted tight junctions using localization microscopy.

    Directory of Open Access Journals (Sweden)

    Rainer Kaufmann

    Full Text Available Tight Junctions (TJ regulate paracellular permeability of tissue barriers. Claudins (Cld form the backbone of TJ-strands. Pore-forming claudins determine the permeability for ions, whereas that for solutes and macromolecules is assumed to be crucially restricted by the strand morphology (i.e., density, branching and continuity. To investigate determinants of the morphology of TJ-strands we established a novel approach using localization microscopy.TJ-strands were reconstituted by stable transfection of HEK293 cells with the barrier-forming Cld3 or Cld5. Strands were investigated at cell-cell contacts by Spectral Position Determination Microscopy (SPDM, a method of localization microscopy using standard fluorophores. Extended TJ-networks of Cld3-YFP and Cld5-YFP were observed. For each network, 200,000 to 1,100,000 individual molecules were detected with a mean localization accuracy of ∼20 nm, yielding a mean structural resolution of ∼50 nm. Compared to conventional fluorescence microscopy, this strongly improved the visualization of strand networks and enabled quantitative morphometric analysis. Two populations of elliptic meshes (mean diameter <100 nm and 300-600 nm, respectively were revealed. For Cld5 the two populations were more separated than for Cld3. Discrimination of non-polymeric molecules and molecules within polymeric strands was achieved. For both subtypes of claudins the mean density of detected molecules was similar and estimated to be ∼24 times higher within the strands than outside the strands.The morphometry and single molecule information provided advances the mechanistic analysis of paracellular barriers. Applying this novel method to different TJ-proteins is expected to significantly improve the understanding of TJ on the molecular level.

  16. Epigenetic programming of T cells impacts immune reconstitution in hematopoietic stem cell transplant recipients.

    Science.gov (United States)

    Hardy, Kristine; Smith, Corey; Tu, Wen Juan; McCuaig, Robert; Panikkar, Archana; Dasari, Vijayendra; Wu, Fan; Tey, Siok-Keen; Hill, Geoffrey R; Khanna, Rajiv; Rao, Sudha

    2018-03-27

    Immune reconstitution following hematopoietic stem cell transplantation (HSCT) is critical in preventing harmful sequelae in recipients with cytomegalovirus (CMV) infection. To understand the molecular mechanisms underlying immune reconstitution kinetics, we profiled the transcriptome-chromatin accessibility landscape of CMV-specific CD8 + T cells from HCST recipients with different immune reconstitution efficiencies. CMV-specific T cells from HSCT recipients with stable antiviral immunity expressed higher levels of interferon/defense response and cell cycle genes in an interconnected network involving PI3KCG , STAT5B , NFAT , RBPJ , and lower HDAC6 , increasing chromatin accessibility at the enhancer regions of immune and T-cell receptor signaling pathway genes. By contrast, the transcriptional and epigenomic signatures of CMV-specific T cells from HSCT recipients with unstable immune reconstitution showed commonalities with T-cell responses in other nonresolving chronic infections. These signatures included higher levels of EGR and KLF factors that, along with lower JARID2 expression, maintained higher accessibility at promoter and CpG-rich regions of genes associated with apoptosis. Furthermore, epigenetic targeting via inhibition of HDAC6 or JARID2 enhanced the transcription of genes associated with differential responses, suggesting that drugs targeting epigenomic modifiers may have therapeutic potential for enhancing immune reconstitution in HSCT recipients. Taken together, these analyses demonstrate that transcription factors and chromatin modulators create different chromatin accessibility landscapes in T cells of HSCT recipients that not only affect immediate gene expression but also differentially prime cells for responses to additional signals. Epigenetic therapy may be a promising strategy to promote immune reconstitution in HSCT recipients. © 2018 by The American Society of Hematology.

  17. Folding DNA into a Lipid-Conjugated Nanobarrel for Controlled Reconstitution of Membrane Proteins.

    Science.gov (United States)

    Dong, Yuanchen; Chen, Shuobing; Zhang, Shijian; Sodroski, Joseph; Yang, Zhongqiang; Liu, Dongsheng; Mao, Youdong

    2018-02-19

    Building upon DNA origami technology, we introduce a method to reconstitute a single membrane protein into a self-assembled DNA nanobarrel that scaffolds a nanodisc-like lipid environment. Compared with the membrane-scaffolding-protein nanodisc technique, our approach gives rise to defined stoichiometry, controlled sizes, as well as enhanced stability and homogeneity in membrane protein reconstitution. We further demonstrate potential applications of the DNA nanobarrels in the structural analysis of membrane proteins. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. A reliable method for reconstituting thymectomized, lethally irradiated guinea pigs with bone marrow cells

    International Nuclear Information System (INIS)

    Terata, N.; Tanio, Y.; Zbar, B.

    1984-01-01

    The authors developed a reliable method for reconstituting thymectomized, lethally irradiated guinea pigs. Injection of 2.5-10 x 10 7 syngeneic bone marrow cells into adult thymectomized, lethally irradiated guinea pigs produced survival of 46-100% of treated animals. Gentamycin sulfate (5 mg/kg of body weight) for 10 days was required for optimal results. Acidified drinking water (pH 2.5) appeared to be required for optimal results. Thymectomized, lethally irradiated, bone marrow reconstituted ('B') guinea pigs had impaired ability to develop delayed cutaneous hypersensitivity to mycobacterial antigens and cutaneous basophil hypersensitivity to keyhole limpet hemocyanin; proliferative responses to phytohemagglutinin were impaired. (Auth.)

  19. On the Major Dramatic Conftict of Shakespeare's the Merchant of Venice

    Institute of Scientific and Technical Information of China (English)

    蒋文

    2000-01-01

    "The Merchant of Venice", a master piece with humanist idea by William Shakespeare, presents readers a dramatic conflict between friendship and greediness, love and cruelty.This essay illustrates briefly how this conflict is created, developed and dramatized. At the same time, the root causes for this conflict is analysed.

  20. Guiding the Noticing: Using a Dramatic Performance Experience to Promote Tellability in Narrative Writing

    Science.gov (United States)

    Clark, Shanetia

    2012-01-01

    In this article, the author describes her use of dramatic performance to promote tellability in narrative writing within a seventh and eighth grade English and language arts classroom. By experiencing dramatic performance, the students were able to actively and physically perform the writing process: brainstorming, drafting, revising, and editing.…

  1. Modeling attitude towards drug treament: the role of internal motivation, external pressure, and dramatic relief.

    Science.gov (United States)

    Conner, Bradley T; Longshore, Douglas; Anglin, M Douglas

    2009-04-01

    Motivation for change has historically been viewed as the crucial element affecting responsiveness to drug treatment. Various external pressures, such as legal coercion, may engender motivation in an individual previously resistant to change. Dramatic relief may be the change process that is most salient as individuals internalize such external pressures. Results of structural equation modeling on data from 465 drug users (58.9% male; 21.3% Black, 34.2% Hispanic/Latino, and 35.1% White) entering drug treatment indicated that internal motivation and external pressure significantly and positively predicted dramatic relief and that dramatic relief significantly predicted attitudes towards drug treatment: chi (2) = 142.20, df = 100, p relief is also likely to be high. When dramatic relief is high, attitudes towards drug treatment are likely to be positive. The findings indicate that interventions to get individuals into drug treatment should include processes that promote Dramatic Relief. Implications for addictions health services are discussed.

  2. Use of a dialyzable short-chain phospholipid for efficient solubilization and reconstitution of influenza virus envelopes

    NARCIS (Netherlands)

    de Jonge, J; Schoen, P; ter Veer, W; Stegmann, T; Wilschut, J; Huckriede, A

    Virosomes are reconstituted viral envelopes that can serve as vaccines and as vehicles for Cellular delivery of various macromolecules. To further advance the use of virosomes, we developed a novel dialysis procedure for the reconstitution of influenza virus membranes that is easily applicable to

  3. Comparative studies on detergent-assisted apocytochrome b6 reconstitution into liposomal bilayers monitored by Zetasizer instruments.

    Directory of Open Access Journals (Sweden)

    Michał A Surma

    Full Text Available The present paper is a systematic, comparative study on the reconstitution of an apocytochrome b6 purified from a heterologous system using a detergent-free method and reconstitution into liposomes performed using three different detergents: SDS, Triton X-100 and DM, and two methods of detergent removal by dialysis and using Bio-Beads. The product size, its distribution and zeta potential, and other parameters were monitored throughout the process. We found that zeta potential of proteoliposomes is correlated with reconstitution efficiency and, as such, can serve as a quick and convenient quality control for reconstitution experiments. We also advocate using detergent-free protein purification methods as they allow for an unfettered choice of detergent for reconstitution, which is the most crucial factor influencing the final product parameters.

  4. Immune reconstitution in patients with Fanconi anemia after allogeneic bone marrow transplantation.

    Science.gov (United States)

    Perlingeiro Beltrame, Miriam; Malvezzi, Mariester; Bonfim, Carmem; Covas, Dimas Tadeu; Orfao, Alberto; Pasquini, Ricardo

    2014-07-01

    Fanconi anemia is an autosomal recessive or X-linked genetic disorder characterized by bone marrow (BM) failure/aplasia. Failure of hematopoiesis results in depletion of the BM stem cell reservoir, which leads to severe anemia, neutropenia and thrombocytopenia, frequently requiring therapeutic interventions, including hematopoietic stem cell transplantation (HSCT). Successful BM transplantation (BMT) requires reconstitution of normal immunity. In the present study, we performed a detailed analysis of the distribution of peripheral blood subsets of T, B and natural killer (NK) lymphocytes in 23 patients with Fanconi anemia before and after BMT on days +30, +60, +100, +180, +270 and +360. In parallel, we evaluated the effect of related versus unrelated donor marrow as well as the presence of graft-versus-host disease (GVHD). After transplantation, we found different kinetics of recovery for the distinct major subsets of lymphocytes. NK cells were the first to recover, followed by cytotoxic CD8(+) T cells and B cells, and finally CD4(+) helper T cells. Early lymphocyte recovery was at the expense of memory cells, potentially derived from the graft, whereas recent thymic emigrant (CD31(+) CD45RA(+)) and naive CD4(+) or CD8(+) T cells rose only at 6 months after HSCT, in the presence of immunosuppressive GVHD prophylactic agents. Only slight differences were observed in the early recovery of cytotoxic CD8(+) T cells among those cases receiving a graft from a related donor versus an unrelated donor. Patients with GVHD displayed a markedly delayed recovery of NK cells and B cells as well as of regulatory T cells and both early thymic emigrant and total CD4(+) T cells. Our results support the utility of post-transplant monitoring of a peripheral blood lymphocyte subset for improved follow-up of patients with Fanconi anemia undergoing BMT. Copyright © 2014 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  5. Arrangement and method for attaching and reattaching a top nozzle in a reconstitutable nuclear fuel assembly

    International Nuclear Information System (INIS)

    Wilson, J.F.; Gjertsen, R.K.; Mayers, J.B.

    1987-01-01

    This patent describes a reconstitutable fuel assembly having at least one control rod guide thimble and a top nozzle. The guide thimble includes an upper end portion. The top nozzle includes a lower adapter plate and an upper hold-down plate. The lower and upper plates have respectively an opening and passageway defined therethrough and aligned with one another. The opening in the lower plate receiving the guide thimble therethrough with its upper end portion extends above the lower plate and towards the passageway of the upper plate. An improved arrangement for mounting the top nozzle on the guide thimble is described comprising: (a) alignment means extending between the plates and receiving the guide thimble upper end portion, (b) complementary means formed on and interconnecting the alignment means and the guide thimble upper end portion so as to connect the alignment means and the guide thimble together. (c) The complementary means further include a secondary interior annular groove formed on the alignment means at a location spaced below the primary annular groove; (d) The complementary means still further include a primary interior section on the alignment means which contains the primary annular groove and a secondary interior section on the alignment means which contains the secondary annular groove. The secondary section is disposed below the primary section and has an interior diameter larger than that of the primary section for facilitating receiving of the severed guide thimble upper end portion back into the alignment means for reconnection of the alignment means and the severed guide thimble together

  6. VH repertoire in progeny of long term lymphoid-cultured cells used to reconstitute immunodeficient mice

    International Nuclear Information System (INIS)

    Denis, K.A.; Timson, L.K.; Witte, O.N.

    1989-01-01

    VH gene utilization in the progeny of long term lymphoid-cultured cells used for reconstitution of severe combined immunodeficient mice under varying conditions was determined. Hybridomas made from the spleens of these animals were evaluated for clonality and donor origin and a panel of 146 independent hybridomas were subsequently examined for VH expression. Hybridomas derived from the spleens of SCID mice reconstituted with fresh cells, used as a control, utilized VH families in proportion to their numerical representation in the genome. However, hybridomas from the spleens of mice reconstituted with long term cultured cells utilized a predominance of the two VH gene families most proximal to JH, characteristic of cells early in B lymphocyte development. Coinjection of thymocytes with cultured fetal liver cells, to provide good levels of T lymphocytes, did not alter this pattern of VH utilization. Irradiation (3 Gy) of the mice before cultured cell injection, which leads to more complete reconstitution of the B cell compartment, was effective in removing this bias in the VH repertoire. Hybridomas derived from these mice expressed their VH genes more in proportion to family size, characteristic of cells later in B lymphocyte development. In this manner, long term lymphoid-cultured cells can be used to study the transitions that occur in VH repertoire expression which appear to be mediated by either B lymphocyte developmental microenvironment or population size

  7. Problems of Equity in the Reconstituted Family: A Social Exchange Analysis.

    Science.gov (United States)

    Nelson, Margaret; Nelson, Gordon K.

    1982-01-01

    Applies social exchange principles to the difficulties of setting up a stepfamily. Discusses obstacles to role adjustment and maintenance of equity among members. Concludes that if the reconstituting family can establish a basis of trust, the stepfamily can merge as a developmental unit toward expansion and commitment. (Author)

  8. Differences in School Behavior and Achievement between Children from Intact, Reconstituted, and Single-Parent Families.

    Science.gov (United States)

    Featherstone, Darin R.; And Others

    1992-01-01

    Analyzed differences in school behavior and achievement among students (n=530) in grades six through nine from intact, reconstituted, and single-parent families. Students from intact, two-parent families had fewer absences and tardies, higher grade point averages, and fewer negative and more positive teacher behavioral ratings than did those from…

  9. Administration of phosphatidylcholine-cholesterol liposomes partially reconstitutes fat absorption in chronically bile-diverted rats

    NARCIS (Netherlands)

    Nishioka, T; Havinga, R; Tazuma, S; Stellaard, F; Kuipers, F; Verkade, HJ

    2004-01-01

    Background and aims: Intestinal bile deficiency in cholestatic patients leads to fat malabsorption. We addressed the potency of model bile, bile salts and phosphatidylcholine (PC)-cholesterol (CH) liposomes to reconstitute fat absorption in permanently bile-diverted (BD) rats. Methods: The plasma

  10. Leprosy reversal reaction as immune reconstitution inflammatory syndrome in patients with AIDS

    OpenAIRE

    Batista, Mariana Dias [UNIFESP; Porro, Adriana Maria [UNIFESP; Maeda, Solange Miki [UNIFESP; Gomes, Elimar Elias [UNIFESP; Yoshioka, Marcia Cristina Naomi [UNIFESP; Enokihara, Mílvia Maria Simões e Silva [UNIFESP; Tomimori, Jane [UNIFESP

    2008-01-01

    We report 2 instances in which reactional borderline leprosy manifested itself as an immune reconstitution phenomenon in patients with acquired immunodeficiency syndrome. We discuss the clinical, laboratory-based, histopathologic, and immunohistochemical characteristics of both patients. Furthermore, we review similar reports from the literature.

  11. Leprosy reversal reaction as immune reconstitution inflammatory syndrome in patients with AIDS.

    Science.gov (United States)

    Batista, Mariana D; Porro, Adriana M; Maeda, Solange M; Gomes, Elimar E; Yoshioka, Márcia C N; Enokihara, Mílvia M S S; Tomimori, Jane

    2008-03-15

    We report 2 instances in which reactional borderline leprosy manifested itself as an immune reconstitution phenomenon in patients with acquired immunodeficiency syndrome. We discuss the clinical, laboratory-based, histopathologic, and immunohistochemical characteristics of both patients. Furthermore, we review similar reports from the literature.

  12. Single biosensor immunoassay for the detection of five aminoglycosides in reconstituted skimmed milk

    NARCIS (Netherlands)

    Haasnoot, W.; Cazemier, G.; Koets, M.; Amerongen, van A.

    2003-01-01

    The application of an optical biosensor (Biacore 3000), with four flow channels (Fcs), in combination with a mixture of four specific antibodies resulted in a competitive inhibition biosensor immunoassay (BIA) for the simultaneous detection of the five relevant aminoglycosides in reconstituted

  13. Immune reconstitution syndrome presenting as probable AIDS-related lymphoma: a case report

    DEFF Research Database (Denmark)

    Mortensen, Bo K; Nielsen, Susanne D; Christensen, Charlotte

    2011-01-01

    ABSTRACT: We report an unusual case of HIV-related immune reconstitution inflammatory syndrome, presenting as suspected AIDS-related lymphoma. Symptoms, initial investigations including fine-needle biopsy and 18F-FDG PET/CT scan were highly compatible with high grade AIDS-related lymphoma, however...

  14. Reconstitution of the NF1 GAP-related domain in NF1-deficient human Schwann cells

    International Nuclear Information System (INIS)

    Thomas, Stacey L.; Deadwyler, Gail D.; Tang, Jun; Stubbs, Evan B.; Muir, David; Hiatt, Kelly K.; Clapp, D. Wade; De Vries, George H.

    2006-01-01

    Schwann cells derived from peripheral nerve sheath tumors from individuals with Neurofibromatosis Type 1 (NF1) are deficient for the protein neurofibromin, which contains a GAP-related domain (NF1-GRD). Neurofibromin-deficient Schwann cells have increased Ras activation, increased proliferation in response to certain growth stimuli, increased angiogenic potential, and altered cell morphology. This study examined whether expression of functional NF1-GRD can reverse the transformed phenotype of neurofibromin-deficient Schwann cells from both benign and malignant peripheral nerve sheath tumors. We reconstituted the NF1-GRD using retroviral transduction and examined the effects on cell morphology, growth potential, and angiogenic potential. NF1-GRD reconstitution resulted in morphologic changes, a 16-33% reduction in Ras activation, and a 53% decrease in proliferation in neurofibromin-deficient Schwann cells. However, NF1-GRD reconstitution was not sufficient to decrease the in vitro angiogenic potential of the cells. This study demonstrates that reconstitution of the NF1-GRD can at least partially reverse the transformation of human NF1 tumor-derived Schwann cells

  15. Post-reconstitution Stability of Telavancin with Commonly Used Diluents and Intravenous Infusion Solutions

    Directory of Open Access Journals (Sweden)

    Zhengtian Gu, PhD

    2015-12-01

    Conclusions: These results are supportive of a total hold time for reconstituted telavancin in vials plus the time in IV infusion solutions in polyvinyl chloride bags to not exceed 12 hours under ambient conditions and 7 days under refrigeration.

  16. Stochastic Threshold Exponential (TE) Model for Hematopoietic Tissue Reconstitution Deficit after Radiation Damage.

    Science.gov (United States)

    Scott, B R; Potter, C A

    2014-07-01

    Whole-body exposure to large radiation doses can cause severe loss of hematopoietic tissue cells and threaten life if the lost cells are not replaced in a timely manner through natural repopulation (a homeostatic mechanism). Repopulation to the baseline level N 0 is called reconstitution and a reconstitution deficit (repopulation shortfall) can occur in a dose-related and organ-specific manner. Scott et al. (2013) previously introduced a deterministic version of a threshold exponential (TE) model of tissue-reconstitution deficit at a given follow-up time that was applied to bone marrow and spleen cellularity (number of constituent cells) data obtained 6 weeks after whole-body gamma-ray exposure of female C.B-17 mice. In this paper a more realistic, stochastic version of the TE model is provided that allows radiation response to vary between different individuals. The Stochastic TE model is applied to post gamma-ray-exposure cellularity data previously reported and also to more limited X-ray cellularity data for whole-body irradiated female C.B-17 mice. Results indicate that the population average threshold for a tissue reconstitution deficit appears to be similar for bone marrow and spleen and for 320-kV-spectrum X-rays and Cs-137 gamma rays. This means that 320-kV spectrum X-rays could successfully be used in conducting such studies.

  17. Functional Reconstitution of a Fungal Natural Product Gene Cluster by Advanced Genome Editing

    DEFF Research Database (Denmark)

    Weber, Jakob; Valiante, Vito; Nødvig, Christina Spuur

    2017-01-01

    is not produced among different isolates. Combining computational analysis with targeted gene editing, we could link a single nucleotide insertion in the polyketide synthase of the trypacidin biosynthetic pathway and reconstitute its production in a nonproducing strain. Thus, we present a CRISPR/Cas9-based tool...... for advanced molecular genetic studies in filamentous fungi, exploiting selectable markers separated from the edited locus....

  18. The role of monocytes in the development of Tuberculosis-associated Immune Reconstitution Inflammatory Syndrome

    NARCIS (Netherlands)

    Tran, Huyen Thi Thanh; van den Bergh, Rafael; Vu, Trung Nghia; Laukens, Kris; Worodria, William; Loembé, Marguerite Massinga; Colebunders, Robert; Kestens, Luc; de Baetselier, Patrick; Raes, Geert; Mayanja-Kizza, Harriet; Mascart, Françoise; den Bergh, Rafaelvan; Locht, Camille; Reiss, Peter; Cobelens, Frank; Ondoa, Pascale; Pakker, Nadine; Mugerwa, Roy

    2014-01-01

    Tuberculosis-associated Immune Reconstitution Inflammatory Syndrome (TB-IRIS) is a common complication of combined antiretroviral therapy (cART) in HIV-TB co-infected patients. However, the disease mechanism is poorly understood, prognosis of TB-IRIS is currently impossible, and diagnosis is highly

  19. B-lymphocyte differentiation in lethally irradiated and reconstituted mice. II. Recovery of humoral immune responsiveness

    International Nuclear Information System (INIS)

    Rozing, J.; Brons, N.H.C.; Benner, R.

    1977-01-01

    The recovery of humoral immune responsiveness was studied in lethally irradiated, fetal liver-reconstituted mice. By means of both membrane fluorescence and antibody formation to sheep red blood cells (SRBC) as a functional assay, the rate of recovery of the compartments of B and T lymphocytes was determined in various lymphoid organs. The recovery of the immunoglobulin-positive (B) cell compartment after irradiation and reconstitution started in the spleen. This organ was also found to be the first in which the recovery of the B-cell population was completed. The interval between the recovery of the B-cell population in the spleen and that in the other organs tested was found to increase when the irradiated mice were reconstituted with spleen colony cells instead of fetal liver cells. This proved to be caused by the number and nature of the reconstituting hemopoietic stem cells. The immunoglobulin-positive (B) cells were found to appear before SRBC-reactive B cells could be demonstrated in spleen, lymph nodes, and Peyer's patches. The appearance of T lymphocytes in the various lymphoid organs required even more time. By means of cell transfer experiments, a sequential appearance of the precursors of anti-SRBC IgM-, IgG-, and IgA-plaque-forming cells could be demonstrated in spleen, bone marrow, lymph nodes, and Peyer's patches

  20. Kinetic Analysis of Lactose Exchange in Proteoliposomes Reconstituted with Purified lac Permease

    NARCIS (Netherlands)

    Lolkema, Julius S.; Carrasco, Nancy; Kaback, H. Ronald

    1991-01-01

    Lactose exchange catalyzed by purified lac permease reconstituted into proteoliposomes was analyzed with unequal concentrations of lactose on either side of the membrane and at low pH so as to prevent equilibration of the two pools. Exchange with external concentrations below 1.0 mM is a

  1. Reconstitution of immunodeficient SCID/beige mice with human cells: Applications in preclinical studies

    International Nuclear Information System (INIS)

    Thomsen, Mogens; Galvani, Sylvain; Canivet, Cindy; Kamar, Nassim; Boehler, Torsten

    2008-01-01

    Experimental studies of the in vivo behaviour of human cells and tissues have become possible with the development of immunodeficient mice strains. Such mice accept readily allogeneic or xenogeneic grafts, including grafts of human cells or tissues, without rejection. In this review we describe different immunodeficient mouse strains that have been used for reconstitution by human immune cells. We subsequently go through the experience that we and others have had with reconstitution, and mention the adverse effects, in particular xenogeneic graft versus host reactions. The use of haematopoietic stem cells avoids such reactions but the immunological reconstitution may take several months. We then report the use of immunodeficient mice for the study of chronic vascular rejection of human mesenteric arteries due to cellular or humoral alloreaction. We have shown that SCID/beige mice grafted with a human artery at the place of the aorta developed a thickening of the intima of the human artery after 5-6 weeks, when they were reconstituted with spleen cells from another human donor. The thickening is mainly due to a proliferation of smooth muscle cells. The same type of lesion developed if they received injection of antibodies towards HLA class I antigens. The arteries of the mouse did not develop any lesion. The arterial lesions closely resembled those seen after clinical organ transplantation. Mice that received spleen cells from the same human donor developed little or no lesions. An important aspect of this experimental transplantation model is the possibility to test drugs that may be used in clinical transplantation. In recent experiments we have shown that novel immunosuppressive drugs can inhibit the hyperproliferation of smooth muscle cells in vitro. Preclinical testing in reconstituted SCID/beige mice grafted with human arteries will permit the evaluation of the potential use of these drugs to prevent chronic vascular rejection. The model also allows

  2. Exploring Lyric, Epic, and Dramatic Voices: Stages of Incandescence in the Poetry of the Aged.

    Science.gov (United States)

    Reed, M. Ann

    1992-01-01

    Identifies true relationships between the psyche and the lyric, epic, and dramatic voices of poetry. Shows how the acts of identifying, responding to, and composing in these three voices engage healing, inspiration, and active imagination among the aging. (SR)

  3. Effect of specialized combined strains on reconstituted milk reduced ...

    African Journals Online (AJOL)

    Yomi

    2012-01-16

    Jan 16, 2012 ... improving the quality of low fat cheese, for example, ... independent replicates of cheese manufacturing were performed on different days. ..... Streptococcus thermophilus in the manufacture of Mexican Panela cheese.

  4. Solubilization, partial purification, and reconstitution of glutamate- and N-methyl-D-aspartate-activated cation channels from brain synaptic membranes

    International Nuclear Information System (INIS)

    Ly, A.M.; Michaelis, E.K.

    1991-01-01

    L-Glutamate-activated cation channel proteins from rat brain synaptic membranes were solubilized, partially purified, and reconstituted into liposomes. Optimal conditions for solubilization and reconstitution included treatment of the membranes with nonionic detergents in the presence of neutral phospholipids plus glycerol. Quench-flow procedures were developed to characterize the rapid kinetics of ion flux induced by receptor agonists. [ 14 C]Methylamine, a cation that permeates through the open channel of both vertebrate and invertebrate glutamate receptors, was used to measure the activity of glutamate receptor-ion channel complexes in reconstituted liposomes. L-Glutamate caused an increase in the rate of [ 14 C]methylamine influx into liposomes reconstituted with either solubilized membrane proteins or partially purified glutamate-binding proteins. Of the major glutamate receptor agonists, only N-methyl-D-aspartate activated cation fluxes in liposomes reconstituted with glutamate-binding proteins. In liposomes reconstituted with glutamate-binding proteins, N-methyl-D-aspartate- or glutamate-induced influx of NA + led to a transient increase in the influx of the lipid-permeable anion probe S 14 CN - . These results indicate the functional reconstitution of N-methyl-D-aspartate-sensitive glutamate receptors and the role of the ∼69-kDa protein in the function of these ion channels

  5. Dramatic Response with Single-Agent Ibrutinib in Multiply Relapsed Marginal Zone Lymphoma with MYD88L265P Mutation

    Directory of Open Access Journals (Sweden)

    Ryan C. Lynch

    2017-09-01

    Full Text Available The B-cell receptor signaling pathway is important in the lymphomagenesis of many lymphomas, including marginal zone lymphoma (MZL. Herein we describe a case of extranodal MZL refractory to multiple lines of therapy. The presence of an IgM paraprotein prompted further evaluation, and the patient was found to have an MYD88L265P mutation. Treatment with ibrutinib led to a dramatic response with prompt resolution of symptoms and significant improvement in measurable sites of disease. The excellent response to ibrutinib in our patient with MYD88L265P-mutated refractory MZL supports a biological rationale for its use.

  6. Borderline tuberculoid leprosy: A manifestation of immune reconstitution inflammatory syndrome in a human immunodeficiency virus infected person

    Directory of Open Access Journals (Sweden)

    Mukhopadhyay Partha

    2006-01-01

    Full Text Available Immune reconstitution inflammatory syndrome describes a collection of inflammatory disorders associated with paradoxical deterioration of various pre-existing processes following start of highly active antiretroviral therapy (HAART in human immunodeficiency virus (HIV-infected patients. Leprosy as an opportunistic infection in immune reconstitution syndrome has been rarely reported in literature. A case of a 30-year-old HIV positive man with extrapulmonary tuberculosis of left foot on HAART having developed borderline tuberculoid leprosy as opportunistic infection in immune reconstitution syndrome has been reported.

  7. B-lymphocyte reconstitution after repeated rituximab treatment in a child with steroid-dependent autoimmune hemolytic anemia

    Directory of Open Access Journals (Sweden)

    Annelieke A.A. van der Linde

    2011-12-01

    Full Text Available We report the detailed long-term reconstitution of B-lymphocyte subpopulations, immunoglobulins, and specific antibody production after two courses of rituximab in a young, previously healthy girl with steroid-dependent autoimmune hemolytic anemia. B-lymphocyte subpopulations were surprisingly normal directly after reconstitution. However, there was a slower reconstitution after the second rituximab course, especially of non-switched and switched memory B-lymphocytes, and a temporary decline in IgM below age-matched reference values.

  8. [Double Endobutto reconstituting coracoclavicular ligament combined with repairing acromioclavicular ligament at stage I for the treatment of acromioclavicular dislocation with Rockwood type III - V].

    Science.gov (United States)

    Hu, Wen-yue; Yu, Chong; Huang, Zhong-ming; Han, Lei

    2015-06-01

    To explore clinical efficacy of double Endobutto reconstituting coracoclavicular ligament combined with repairing acromioclavicular ligament in stage I in treating acromioclavicular dislocation with Rockwood type III - V . From January 2010 to September 2013, 56 patients with Rockwood type III - V acromioclavicular dislocation were treated by operation, including 20 males and 36 femlaes, aged from 32 to 52 years old with an average of 38.5 years old. Twenty-five patients were on the left side and 31 cases on the right side. The time from injury to operation was from 3 to 14 days, averaged 7 days. All patients were diagnosed as acromioclavicular dislocation with Rockwood type III - V, and double Endobutto were used to reconstituting coracoclavicular ligament, line metal anchors were applied for repairing acromioclavicular ligament. Postoperative complications were observed, Karlsson and Constant-Murley evaluation standard were used to evaluate clinical effects. All patients were followed up from 8 to 24 months with average of 11 months. According to Karlsson evaluation standard at 6 months after operation,42 cases were grade A, 13 were grade B and 1 was grade C. Constant-Murley score were improved from (42.80±5.43) before operation to (91.75±4.27) at 6 months after operation. All items at 6 months after operation were better than that of preoperative items. Forty-eight patients got excellent results, 7 were moderate and only 1 with bad result. No shoulder joint adhesion, screw loosening or breakage were occurred during following up. Double Endobutto reconstituting coracoclavicular ligament combined with repairing acromioclavicular ligament in stage I for the treatment of acromioclavicular dislocation with Rockwood type III - V could obtain early staisfied clinical effects, and benefit for early recovery of shoulder joint function.

  9. Permeability enhancers dramatically increase zanamivir absolute bioavailability in rats: implications for an orally bioavailable influenza treatment.

    Directory of Open Access Journals (Sweden)

    Eric H Holmes

    Full Text Available We have demonstrated that simple formulations composed of the parent drug in combination with generally regarded as safe (GRAS permeability enhancers are capable of dramatically increasing the absolute bioavailability of zanamivir. This has the advantage of not requiring modification of the drug structure to promote absorption, thus reducing the regulatory challenges involved in conversion of an inhaled to oral route of administration of an approved drug. Absolute bioavailability increases of up to 24-fold were observed when Capmul MCM L8 (composed of mono- and diglycerides of caprylic/capric acids in glycerol was mixed with 1.5 mg of zanamivir and administered intraduodenally to rats. Rapid uptake (t(max of 5 min and a C(max of over 7200 ng/mL was achieved. Variation of the drug load or amount of enhancer demonstrated a generally linear variation in absorption, indicating an ability to optimize a formulation for a desired outcome such as a targeted C(max for enzyme saturation. No absorption enhancement was observed when the enhancer was given 2 hr prior to drug administration, indicating, in combination with the observed tmax, that absorption enhancement is temporary. This property is significant and aligns well with therapeutic applications to limit undesirable drug-drug interactions, potentially due to the presence of other poorly absorbed polar drugs. These results suggest that optimal human oral dosage forms of zanamivir should be enteric-coated gelcaps or softgels for intraduodenal release. There continues to be a strong need and market for multiple neuraminidase inhibitors for influenza treatment. Creation of orally available formulations of inhibitor drugs that are currently administered intravenously or by inhalation would provide a significant improvement in treatment of influenza. The very simple GRAS formulation components and anticipated dosage forms would require low manufacturing costs and yield enhanced convenience. These results

  10. Reconstitution of FMN-free NADPH-cytochrome P-450 reductase with a phosphorothioate analog of FMN: 31P NMR studies of the reconstituted protein

    International Nuclear Information System (INIS)

    Krum, D.P.; Otvos, J.D.; Calhoun, J.P.; Miziorko, H.M.; Masters, B.S.S.

    1987-01-01

    A phosphorothioate analog of FMN (FMNS) has been synthesized and shown to be completely competent in reconstituting the FMN-free form of NADPH-cytochrome P-450 reductase as evidenced by flavin determinations and cytochrome c reductase activity assays. The FMNS-reconstituted FMN-free reductase gives rise to an air-stable semiquinone, and the fluorescence of FMNS is quenched upon addition of FMN-free reductase. 31 P NMR spectra of the FMN-free reductase reveal only two resonances (-7.3 and -11.3 ppm), which are attributable to FAD. This result confirms the assignments of Otvos et al, and demonstrates unequivocally that there are no phosphate residues other than those of FMN and FAD attached to the steapsin-solubilized reductase. The addition of FMN to the FMN-free reductase resulted in the appearance of one additional resonance at 3.9 ppm. Addition of FMNS to the FMN-free reductase caused no change, surprisingly, in the 31 P NMR spectrum until Mn(II) was added, after which a peak centered at ∼ 45 ppm was observed. This unexpected result may be explained if the T 1 for the phosphate of FMNS is significantly longer than that of FMN, and suggests that the sulfur atom of FMNS may perturb the interaction of the phosphate with its protein environment. These results demonstrate the utility of phosphorothioate analogs as mechanistic probes for proteins containing nucleotide cofactors

  11. Cellulose ionics: switching ionic diode responses by surface charge in reconstituted cellulose films.

    Science.gov (United States)

    Aaronson, Barak D B; Wigmore, David; Johns, Marcus A; Scott, Janet L; Polikarpov, Igor; Marken, Frank

    2017-09-25

    Cellulose films as well as chitosan-modified cellulose films of approximately 5 μm thickness, reconstituted from ionic liquid media onto a poly(ethylene-terephthalate) (PET, 6 μm thickness) film with a 5, 10, 20, or 40 μm diameter laser-drilled microhole, show significant current rectification in aqueous NaCl. Reconstituted α-cellulose films provide "cationic diodes" (due to predominant cation conductivity) whereas chitosan-doped cellulose shows "anionic diode" effects (due to predominant anion conductivity). The current rectification, or "ionic diode" behaviour, is investigated as a function of NaCl concentration, pH, microhole diameter, and molecular weight of the chitosan dopant. Future applications are envisaged exploiting the surface charge induced switching of diode currents for signal amplification in sensing.

  12. Reconstitution of Qbeta RNA replicase from a covalently bonded elongation factor Tu-Ts complex

    DEFF Research Database (Denmark)

    Brown, S; Blumenthal, T

    1976-01-01

    of these polypeptides, protein synthesis elongation factors EF-Tu and EF-Ts, can be covalently crosslinked with dimethyl suberimidate to form a complex which lacks the ability to catalyze the known host functions catalyzed by the individual elongation factors. Using a previously developed reconstitution system we have...... examined the effects of crosslinking the EF-Tu-Ts complex on reconstituted replicase activity. Renaturation is significantly more efficient when exogenously added native EF-Tu-Ts is crosslinked than when it is not. Crosslinked EF-Tu-Ts can be purified from a crude crosslinked postribosomal supernatant...... by its ability to replace EF-Tu and EF-Ts in the renaturation of denatured Qbeta replicase. A sample of Qbeta replicase with crosslinked EF-Tu-Ts replacing the individual elongation factors was prepared. Although it lacked EF-Tu and EF-Ts activities, it could initiate transcription of both poly...

  13. Inactivation of Nondesiccated and Desiccated Cronobacter sakazakii in Reconstituted Infant Formula by Combination of Citral and Mild Heat.

    Science.gov (United States)

    Shi, Chao; Jia, Zhenyu; Sun, Yi; Chen, Yifei; Guo, Du; Liu, Zhiyuan; Wen, Qiwu; Guo, Xiao; Ma, Linlin; Yang, Baowei; Baloch, Allah Bux; Xia, Xiaodong

    2017-07-01

    The objective of this study was to evaluate the combined effect of citral plus mild heat on nondesiccated and desiccated Cronobacter sakazakii in reconstituted infant formula. Various concentrations of citral (0, 0.3, 0.6, and 0.9%) combined with various temperatures (25, 45, 50, and 55°C) were applied to nondesiccated and desiccated cocktails of three C. sakazakii strains (approximately 6.0 log CFU mL -1 ) in reconstituted infant formula, and the bacterial populations were assayed periodically. The combined treatments had marked antimicrobial effects on C. sakazakii compared with the control. Desiccated cells were more susceptible to citral than were nondesiccated cells in reconstituted infant formula. These findings suggest there is a potential application of citral in combination with mild heat to control C. sakazakii during preparation of reconstituted infant formula.

  14. Defense Logistics: Army Has Not Fully Planned or Budgeted for the Reconstitution of Its Afloat Prepositioned Stocks

    National Research Council Canada - National Science Library

    Solis, William M

    2008-01-01

    ...) the extent to which the Army's APS reconstitution strategy is reflected in current defense budget requests and cost estimates for restoring the prepositioned equipment sets to a posture that fully...

  15. A bio-ecological interpretation of the relationship challenges in the context of the reconstituted family

    OpenAIRE

    Ebersohn, S; Bouwer, AC

    2015-01-01

    From an educational psychology perspective, family life - as a child's primary educational situation - is changing drastically as divorces increase worldwide. Various challenges to relationships accompany the restructuring of family systems after divorce. When remarriage occurs, children's shared membership of two family microsystems and the resultant complexity of the mesosystem cause the reconstituted family situation to come to differ radically from that of a nuclear family. The purpose of...

  16. Liquidity, Reconstitution, and the Value of U.S. Treasury Strips.

    OpenAIRE

    Daves, Phillip R; Ehrhardt, Michael C

    1993-01-01

    An apparent pricing anomaly exists in the market for U.S. Treasury strips: zero-coupon strips created from principal payments typically trade at significantly higher prices than otherwi se identical zero-coupon strips created from coupon payments. In additi on to documenting this phenomenon, this study demonstrates that differences in liquidity and differences in reconstitution characteristics explain much of this price variation. Copyright 1993 by American Finance Association.

  17. Immune reconstitution syndrome in a human immunodeficiency virus infected child due to giardiasis leading to shock

    Directory of Open Access Journals (Sweden)

    Sneha Nandy

    2015-01-01

    Full Text Available Human immunodeficiency virus (HIV-associated immune reconstitution inflammatory syndrome has been reported in association with tuberculosis, herpes zoster (shingles, Cryptococcus neoformans, Kaposi′s sarcoma, Pneumocystis pneumonia, hepatitis B virus, hepatitis C virus, herpes simplex virus, Histoplasma capsulatum, human papillomavirus, and Cytomegalovirus. However, it has never been documented with giardiasis. We present a 7-year-old HIV infected girl who developed diarrhea and shock following the initiation of antiretroviral therapy, and her stool showed the presence of giardiasis.

  18. Activation of Saccharomyces cerevisiae Mlh1-Pms1 Endonuclease in a Reconstituted Mismatch Repair System*

    Science.gov (United States)

    Smith, Catherine E.; Bowen, Nikki; Graham, William J.; Goellner, Eva M.; Srivatsan, Anjana; Kolodner, Richard D.

    2015-01-01

    Previous studies reported the reconstitution of an Mlh1-Pms1-independent 5′ nick-directed mismatch repair (MMR) reaction using Saccharomyces cerevisiae proteins. Here we describe the reconstitution of a mispair-dependent Mlh1-Pms1 endonuclease activation reaction requiring Msh2-Msh6 (or Msh2-Msh3), proliferating cell nuclear antigen (PCNA), and replication factor C (RFC) and a reconstituted Mlh1-Pms1-dependent 3′ nick-directed MMR reaction requiring Msh2-Msh6 (or Msh2-Msh3), exonuclease 1 (Exo1), replication protein A (RPA), RFC, PCNA, and DNA polymerase δ. Both reactions required Mg2+ and Mn2+ for optimal activity. The MMR reaction also required two reaction stages in which the first stage required incubation of Mlh1-Pms1 with substrate DNA, with or without Msh2-Msh6 (or Msh2-Msh3), PCNA, and RFC but did not require nicking of the substrate, followed by a second stage in which other proteins were added. Analysis of different mutant proteins demonstrated that both reactions required a functional Mlh1-Pms1 endonuclease active site, as well as mispair recognition and Mlh1-Pms1 recruitment by Msh2-Msh6 but not sliding clamp formation. Mutant Mlh1-Pms1 and PCNA proteins that were defective for Exo1-independent but not Exo1-dependent MMR in vivo were partially defective in the Mlh1-Pms1 endonuclease and MMR reactions, suggesting that both reactions reflect the activation of Mlh1-Pms1 seen in Exo1-independent MMR in vivo. The availability of this reconstituted MMR reaction should now make it possible to better study both Exo1-independent and Exo1-dependent MMR. PMID:26170454

  19. Activation of Saccharomyces cerevisiae Mlh1-Pms1 Endonuclease in a Reconstituted Mismatch Repair System.

    Science.gov (United States)

    Smith, Catherine E; Bowen, Nikki; Graham, William J; Goellner, Eva M; Srivatsan, Anjana; Kolodner, Richard D

    2015-08-28

    Previous studies reported the reconstitution of an Mlh1-Pms1-independent 5' nick-directed mismatch repair (MMR) reaction using Saccharomyces cerevisiae proteins. Here we describe the reconstitution of a mispair-dependent Mlh1-Pms1 endonuclease activation reaction requiring Msh2-Msh6 (or Msh2-Msh3), proliferating cell nuclear antigen (PCNA), and replication factor C (RFC) and a reconstituted Mlh1-Pms1-dependent 3' nick-directed MMR reaction requiring Msh2-Msh6 (or Msh2-Msh3), exonuclease 1 (Exo1), replication protein A (RPA), RFC, PCNA, and DNA polymerase δ. Both reactions required Mg(2+) and Mn(2+) for optimal activity. The MMR reaction also required two reaction stages in which the first stage required incubation of Mlh1-Pms1 with substrate DNA, with or without Msh2-Msh6 (or Msh2-Msh3), PCNA, and RFC but did not require nicking of the substrate, followed by a second stage in which other proteins were added. Analysis of different mutant proteins demonstrated that both reactions required a functional Mlh1-Pms1 endonuclease active site, as well as mispair recognition and Mlh1-Pms1 recruitment by Msh2-Msh6 but not sliding clamp formation. Mutant Mlh1-Pms1 and PCNA proteins that were defective for Exo1-independent but not Exo1-dependent MMR in vivo were partially defective in the Mlh1-Pms1 endonuclease and MMR reactions, suggesting that both reactions reflect the activation of Mlh1-Pms1 seen in Exo1-independent MMR in vivo. The availability of this reconstituted MMR reaction should now make it possible to better study both Exo1-independent and Exo1-dependent MMR. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. The effect of interrupted anti-retroviral treatment on the reconstitution ...

    African Journals Online (AJOL)

    Objectives: To ascertain the effect of interrupted ART on reconstitution of CD4+ and CD8+ T sub-sets in TB patients. Methods: Participants with HIV (CD4>350 cells/μL) and TB were recruited under a larger phase 3 open label randomised controlled clinical trial. The CD45RO and CD62L markers were measured on CD4+ ...

  1. Acute gouty arthritis as a manifestation of immune reconstitution inflammatory syndrome after initiation of antiretroviral therapy

    Directory of Open Access Journals (Sweden)

    Walter de Araujo Eyer-Silva

    2012-08-01

    Full Text Available Immune reconstitution inflammatory syndrome (IRIS in HIV-infected subjects initiating antiretroviral therapy most commonly involves new or worsening manifestations of previously subclinical or overt infectious diseases. Reports of non-infectious IRIS are much less common but represent important diagnostic and treatment challenges. We report on a 34-year-old HIV-infected male patient with no history of gout who developed acute gouty arthritis in a single joint one month after initiating highly active antiretroviral therapy.

  2. Reconstitution as a Guiding Principle to US Army Force Structure: High Risk or Prudent Hedge?

    Science.gov (United States)

    2014-05-20

    integrated process, does offer the United States a means to mitigate these risks. The United States is uniquely postured to take advantage of reconstitution...Advanced Mili tary Studies Hen Accepted this 22"d day of May 20 14 b irector, Graduate Degree ProQ.ram The op inions and conclusions expressed...to conven th ese resources into actual capability w hen needed req uires a disc iplined approach that accounts for the doctr inal organiza tional

  3. Immune reconstitution syndrome presenting as probable AIDS-related lymphoma: a case report

    OpenAIRE

    Christensen Charlotte B; Nielsen Susanne D; Mortensen Bo K; Helweg-Larsen Jannik

    2011-01-01

    Abstract We report an unusual case of HIV-related immune reconstitution inflammatory syndrome, presenting as suspected AIDS-related lymphoma. Symptoms, initial investigations including fine-needle biopsy and 18F-FDG PET/CT scan were highly compatible with high grade AIDS-related lymphoma, however subsequently IRIS was diagnosed. We discuss pitfalls in the interpretation of diagnostic results in ARL versus IRIS.

  4. Expression, purification and functional reconstitution of slack sodium-activated potassium channels.

    Science.gov (United States)

    Yan, Yangyang; Yang, Youshan; Bian, Shumin; Sigworth, Fred J

    2012-11-01

    The slack (slo2.2) gene codes for a potassium-channel α-subunit of the 6TM voltage-gated channel family. Expression of slack results in Na(+)-activated potassium channel activity in various cell types. We describe the purification and reconstitution of Slack protein and show that the Slack α-subunit alone is sufficient for potassium channel activity activated by sodium ions as assayed in planar bilayer membranes and in membrane vesicles.

  5. Reconstituting development of pancreatic intraepithelial neoplasia from primary human pancreas duct cells

    OpenAIRE

    Lee, Jonghyeob; Snyder, Emily R.; Liu, Yinghua; Gu, Xueying; Wang, Jing; Flowers, Brittany M.; Kim, Yoo Jung; Park, Sangbin; Szot, Gregory L.; Hruban, Ralph H.; Longacre, Teri A.; Kim, Seung K.

    2017-01-01

    Development of systems that reconstitute hallmark features of human pancreatic intraepithelial neoplasia (PanINs), the precursor to pancreatic ductal adenocarcinoma, could generate new strategies for early diagnosis and intervention. However, human cell-based PanIN models with defined mutations are unavailable. Here, we report that genetic modification of primary human pancreatic cells leads to development of lesions resembling native human PanINs. Primary human pancreas duct cells harbouring...

  6. Immune reconstitution inflammatory syndrome unmasking erythema nodosum leprosum: A rare case report

    Directory of Open Access Journals (Sweden)

    Geeta Kiran Arakkal

    2015-01-01

    Full Text Available Immune reconstitution inflammatory syndrome (IRIS occurs as an acute symptomatic expression of a latent infection during the recovery of immune system in response to antiretroviral therapy in HIV patients. IRIS triggers both opportunistic and non-opportunistic infections. We report a case of IRIS in a patient with HIV, presenting as erythema nodosum leprosum (ENL, which led to unmasking of lepromatous leprosy following anti-retroviral therapy (ART.

  7. Immune Reconstitution Inflammatory Syndrome Unmasking or Worsening AIDS-Related Progressive Multifocal Leukoencephalopathy: A Literature Review

    Directory of Open Access Journals (Sweden)

    Anna Fournier

    2017-05-01

    Full Text Available Incidence of progressive multifocal leukoencephalopathy (PML in HIV-infected patients has declined in the combined antiretroviral therapy (cART era although a growing number of acquired immunodeficiency syndrome (AIDS-related PML-immune reconstitution inflammatory syndromes (PML-IRIS have been published during the same period. Therapeutic management of PML-IRIS is not consensual and mainly relies on corticosteroids. Our main aim was, in addition to provide a thoughtful analysis of published PML-IRIS cases, to assess the benefit of corticosteroids in the management of PML-IRIS, focusing on confirmed cases. We performed a literature review of the 46 confirmed cases of PML-IRIS cases occurring in HIV-infected patients from 1998 to September 2016 (21 unmasking and 25 paradoxical PML-IRIS. AIDS-related PML-IRIS patients were mostly men (sex ratio 4/1 with a median age of 40.5 years (range 12–66. Median CD4 T cell count before cART and at PML-IRIS onset was 45/μl (0–301 and 101/μl (20–610, respectively. After cART initiation, PML-IRIS occurred within a median timescale of 38 days (18–120. Clinical signs were motor deficits (69%, speech disorders (36%, cognitive disorders (33%, cerebellar ataxia (28%, and visual disturbances (23%. Brain MRI revealed hyperintense areas on T2-weighted sequences and FLAIR images (76% and suggestive contrast enhancement (87%. PCR for John Cunningham virus (JCV in cerebrospinal fluid (CSF was positive in only 84% of cases; however, when performed, brain biopsy confirmed diagnosis of PML in 90% of cases and demonstrated histological signs of IRIS in 95% of cases. Clinical worsening related to PML-IRIS and leading to death was observed in 28% of cases. Corticosteroids were prescribed in 63% of cases and maraviroc in one case. Statistical analysis failed to demonstrate significant benefit from steroid treatment, despite spectacular improvement in certain cases. Diagnosis of PML-IRIS should be considered in HIV

  8. Immune Reconstitution Inflammatory Syndrome Unmasking or Worsening AIDS-Related Progressive Multifocal Leukoencephalopathy: A Literature Review.

    Science.gov (United States)

    Fournier, Anna; Martin-Blondel, Guillaume; Lechapt-Zalcman, Emmanuèle; Dina, Julia; Kazemi, Apolline; Verdon, Renaud; Mortier, Emmanuel; de La Blanchardière, Arnaud

    2017-01-01

    Incidence of progressive multifocal leukoencephalopathy (PML) in HIV-infected patients has declined in the combined antiretroviral therapy (cART) era although a growing number of acquired immunodeficiency syndrome (AIDS)-related PML-immune reconstitution inflammatory syndromes (PML-IRIS) have been published during the same period. Therapeutic management of PML-IRIS is not consensual and mainly relies on corticosteroids. Our main aim was, in addition to provide a thoughtful analysis of published PML-IRIS cases, to assess the benefit of corticosteroids in the management of PML-IRIS, focusing on confirmed cases. We performed a literature review of the 46 confirmed cases of PML-IRIS cases occurring in HIV-infected patients from 1998 to September 2016 (21 unmasking and 25 paradoxical PML-IRIS). AIDS-related PML-IRIS patients were mostly men (sex ratio 4/1) with a median age of 40.5 years (range 12-66). Median CD4 T cell count before cART and at PML-IRIS onset was 45/μl (0-301) and 101/μl (20-610), respectively. After cART initiation, PML-IRIS occurred within a median timescale of 38 days (18-120). Clinical signs were motor deficits (69%), speech disorders (36%), cognitive disorders (33%), cerebellar ataxia (28%), and visual disturbances (23%). Brain MRI revealed hyperintense areas on T2-weighted sequences and FLAIR images (76%) and suggestive contrast enhancement (87%). PCR for John Cunningham virus (JCV) in cerebrospinal fluid (CSF) was positive in only 84% of cases; however, when performed, brain biopsy confirmed diagnosis of PML in 90% of cases and demonstrated histological signs of IRIS in 95% of cases. Clinical worsening related to PML-IRIS and leading to death was observed in 28% of cases. Corticosteroids were prescribed in 63% of cases and maraviroc in one case. Statistical analysis failed to demonstrate significant benefit from steroid treatment, despite spectacular improvement in certain cases. Diagnosis of PML-IRIS should be considered in HIV

  9. Reconstitution of the yeast RNA polymerase III transcription system with all recombinant factors.

    Science.gov (United States)

    Ducrot, Cécile; Lefebvre, Olivier; Landrieux, Emilie; Guirouilh-Barbat, Josée; Sentenac, André; Acker, Joel

    2006-04-28

    Transcription factor TFIIIC is a multisubunit complex required for promoter recognition and transcriptional activation of class III genes. We describe here the reconstitution of complete recombinant yeast TFIIIC and the molecular characterization of its two DNA-binding domains, tauA and tauB, using the baculovirus expression system. The B block-binding module, rtauB, was reconstituted with rtau138, rtau91, and rtau60 subunits. rtau131, rtau95, and rtau55 formed also a stable complex, rtauA, that displayed nonspecific DNA binding activity. Recombinant rTFIIIC was functionally equivalent to purified yeast TFIIIC, suggesting that the six recombinant subunits are necessary and sufficient to reconstitute a transcriptionally active TFIIIC complex. The formation and the properties of rTFIIIC-DNA complexes were affected by dephosphorylation treatments. The combination of complete recombinant rTFIIIC and rTFIIIB directed a low level of basal transcription, much weaker than with the crude B'' fraction, suggesting the existence of auxiliary factors that could modulate the yeast RNA polymerase III transcription system.

  10. HIV-positive patient with herpes zoster: a manifestation of the immune reconstitution inflammatory syndrome.

    Science.gov (United States)

    Lutwak, Nancy; Dill, Curt

    2012-01-01

    Herpes zoster is a common illness that can lead to serious morbidity. There is now evidence that HIV-infected patients who have been treated with antiretroviral therapy are at greater risk of developing herpes zoster not when they are severely immunocompromised but, paradoxically, when their immune system is recovering. This is a manifestation of the immune reconstitution inflammatory syndrome. The objectives of this report are to (1) inform health care providers that HIV-infected patients may develop multiple infectious, autoimmune, and oncological manifestations after treatment with antiretroviral medication, as they have immune system reconstitution, and (2) discuss herpes zoster, one of the possible manifestations. The patient is a 68-year-old HIV-positive man who presented with herpes zoster after being treated with highly active antiretroviral therapy (HAART) when his immune system was recovering, not when he was most immunosuppressed. Emergency department physicians should be aware that HIV-infected patients treated with HAART may have clinical deterioration despite immune system strengthening. This immune reconstitution inflammatory syndrome can present with infectious, autoimmune, or oncological manifestations. Our case patient, an HIV-positive man with immune system recovery after treatment with HAART, presented with an infectious manifestation, herpes zoster.

  11. Purified reconstituted lac carrier protein from Escherichia coli is fully functional.

    Science.gov (United States)

    Viitanen, P; Garcia, M L; Kaback, H R

    1984-03-01

    Proteoliposomes reconstituted with lac carrier protein purified from the plasma membrane of Escherichia coli catalyze each of the translocation reactions typical of the beta-galactoside transport system (i.e., active transport, counterflow, facilitated influx and efflux) with turnover numbers and apparent Km values comparable to those observed in right-side-out membrane vesicles. Furthermore, detailed kinetic studies show that the reconstituted system exhibits properties analogous to those observed in membrane vesicles. Imposition of a membrane potential (delta psi, interior negative) causes a marked decrease in apparent Km (by a factor of 7 to 10) with a smaller increase in Vmax (approximately equal to 3-fold). At submaximal values of delta psi, the reconstituted carrier exhibits biphasic kinetics, with one component manifesting the kinetic parameters of active transport and the other exhibiting the characteristics of facilitated diffusion. Finally, at low lactose concentrations, the initial velocity of influx varies linearly with the square of the proton electro-chemical gradient. The results provide quantitative support for the contention that a single polypeptide species, the product of the lac y gene, is responsible for each of the transport reactions typical of the beta-galactoside transport system.

  12. Study of 25 cases of exchange transfusion by reconstituted blood in hemolytic disease of newborn

    Science.gov (United States)

    Sharma, D. C.; Rai, Sunita; Mehra, Aakash; Kaur, M. M.; Sao, Satya; Gaur, Ajay; Sapra, Rahul

    2007-01-01

    This study was aimed to review and establish the practice of exchange transfusion (ET) with reconstituted blood in neonates and to observe fall of bilirubin and its comparison with related studies. Twenty-five neonates diagnosed as hemolytic disease of newborn (HDN) were selected for this study, in which exchange transfusion was carried out as one of the treatments for hyperbilirubinemia. Out of the 25 cases, 15 were of Rhesus (Rh) HDN, while ABO and other blood groups constituted 6 and 4 HDN cases respectively. First, the neonates's and mother's blood samples were subjected to relevant investigations. After that, for neonates having Rh HDN, O Rh negative cells suspended in AB plasma were given, O Rh positive cells suspended in AB plasma were given to ABO HDN; and O positive cells, which were indirect Coomb's cross-matched compatible with neonates’ and mother's serum / plasma, suspended in AB plasma were given to the neonates having HDN because of other blood group antibodies. The exchange transfusion (ET) was carried out taking all aseptic precautions by Push-Pull technique with double-volume exchange transfusion method. The average post-exchange fall in serum indirect bilirubin was (52.01%) in all 25 cases, which was found to be more significant than the previous studies. Looking into the superiority of the exchange transfusion in HDN by reconstituted blood, the reconstituted blood can be modified and supplied as per the requirement and conditions. PMID:21938234

  13. Study of 25 cases of exchange transfusion by reconstituted blood in hemolytic disease of newborn

    Directory of Open Access Journals (Sweden)

    Sharma D

    2007-01-01

    Full Text Available This study was aimed to review and establish the practice of exchange transfusion (ET with reconstituted blood in neonates and to observe fall of bilirubin and its comparison with related studies. Twenty-five neonates diagnosed as hemolytic disease of newborn (HDN were selected for this study, in which exchange transfusion was carried out as one of the treatments for hyperbilirubinemia. Out of the 25 cases, 15 were of Rhesus (Rh HDN, while ABO and other blood groups constituted 6 and 4 HDN cases respectively. First, the neonates′ and mother′s blood samples were subjected to relevant investigations. After that, for neonates having Rh HDN, O Rh negative cells suspended in AB plasma were given, O Rh positive cells suspended in AB plasma were given to ABO HDN; and O positive cells, which were indirect Coomb′s cross-matched compatible with neonates′ and mother′s serum / plasma, suspended in AB plasma were given to the neonates having HDN because of other blood group antibodies. The exchange transfusion (ET was carried out taking all aseptic precautions by Push-Pull technique with double-volume exchange transfusion method. The average post-exchange fall in serum indirect bilirubin was (52.01% in all 25 cases, which was found to be more significant than the previous studies. Looking into the superiority of the exchange transfusion in HDN by reconstituted blood, the reconstituted blood can be modified and supplied as per the requirement and conditions.

  14. Human hepatocyte depletion in the presence of HIV-1 infection in dual reconstituted humanized mice

    Science.gov (United States)

    Wang, Weimin; Cheng, Yan; Makarov, Edward; Ganesan, Murali; Gebhart, Catherine L.; Gorantla, Santhi; Osna, Natalia

    2018-01-01

    ABSTRACT Human immunodeficiency virus type 1 (HIV-1) infection impairs liver function, and liver diseases have become a leading cause of morbidity in infected patients. The immunopathology of liver damage caused by HIV-1 remains unclear. We used chimeric mice dually reconstituted with a human immune system and hepatocytes to address the relevance of the model to pathobiology questions related to human hepatocyte survival in the presence of systemic infection. TK-NOG males were transplanted with mismatched human hematopoietic stem/progenitor cells and hepatocytes, human albumin concentration and the presence of human immune cells in blood were monitored for hepatocytes and immune reconstitution, and mice were infected with HIV-1. HIV-1-infected animals showed a decline in human albumin concentration with a significant reduction in percentage of human hepatocytes compared to uninfected mice. The decrease in human albumin levels correlated with a decline in CD4+ cells in the liver and with an increase in HIV-1 viral load. HIV-1 infection elicited proinflammatory response in the immunological milieu of the liver in HIV-infected mice compared to uninfected animals, as determined by upregulation of IL23, CXCL10 and multiple toll-like receptor expression. The inflammatory reaction associated with HIV-1 infection in vivo could contribute to the depletion and dysfunction of hepatocytes. The dual reconstituted TK-NOG mouse model is a feasible platform to investigate hepatocyte-related HIV-1 immunopathogenesis. This article has an associated First Person interview with the first author of the paper. PMID:29361613

  15. Tritium contamination of hematopoietic stem cells alters long-term hematopoietic reconstitution

    International Nuclear Information System (INIS)

    Di Giacomo, F.; Barroca, V.; Laurent, D.; Lewandowski, D.; Saintigny, Y.; Romeo, P.H.; Granotier, Ch.; Boussin, F.D.

    2011-01-01

    Purpose: In vivo effects of tritium contamination are poorly documented. Here, we study the effects of tritiated Thymidine ([ 3 H] Thymidine) or tritiated water (HTO) contamination on the biological properties of hematopoietic stem cells (HSC). Materials and methods: Mouse HSC were contaminated with concentrations of [ 3 H] Thymidine ranging from 0.37-37.03 kBq/ml or of HTO ranging from 5-50 kBq/ml. The biological properties of contaminated HSC were studied in vitro after HTO contamination and in vitro and in vivo after [ 3 H] Thymidine contamination. Results: Proliferation, viability and double-strand breaks were dependent on [ 3 H] Thymidine or HTO concentrations used for contamination but in vitro myeloid differentiation of HSC was not affected by [ 3 H] Thymidine contamination. [ 3 H] Thymidine contaminated HSC showed a compromised long-term capacity of hematopoietic reconstitution and competition experiments showed an up to two-fold decreased capacity of contaminated HSC to reconstitute hematopoiesis. These defects were not due to impaired homing in bone marrow but to an initial decreased proliferation rate of HSC. Conclusion: These results indicate that contaminations of HSC with doses of tritium that do not result in cell death, induce short-term effects on proliferation and cell cycle and long-term effects on hematopoietic reconstitution capacity of contaminated HSC. (authors)

  16. Human hepatocyte depletion in the presence of HIV-1 infection in dual reconstituted humanized mice

    Directory of Open Access Journals (Sweden)

    Raghubendra Singh Dagur

    2018-02-01

    Full Text Available Human immunodeficiency virus type 1 (HIV-1 infection impairs liver function, and liver diseases have become a leading cause of morbidity in infected patients. The immunopathology of liver damage caused by HIV-1 remains unclear. We used chimeric mice dually reconstituted with a human immune system and hepatocytes to address the relevance of the model to pathobiology questions related to human hepatocyte survival in the presence of systemic infection. TK-NOG males were transplanted with mismatched human hematopoietic stem/progenitor cells and hepatocytes, human albumin concentration and the presence of human immune cells in blood were monitored for hepatocytes and immune reconstitution, and mice were infected with HIV-1. HIV-1-infected animals showed a decline in human albumin concentration with a significant reduction in percentage of human hepatocytes compared to uninfected mice. The decrease in human albumin levels correlated with a decline in CD4+ cells in the liver and with an increase in HIV-1 viral load. HIV-1 infection elicited proinflammatory response in the immunological milieu of the liver in HIV-infected mice compared to uninfected animals, as determined by upregulation of IL23, CXCL10 and multiple toll-like receptor expression. The inflammatory reaction associated with HIV-1 infection in vivo could contribute to the depletion and dysfunction of hepatocytes. The dual reconstituted TK-NOG mouse model is a feasible platform to investigate hepatocyte-related HIV-1 immunopathogenesis. This article has an associated First Person interview with the first author of the paper.

  17. A bio-ecological interpretation of the relationship challenges in the context of the reconstituted family

    Directory of Open Access Journals (Sweden)

    S Ebersohn

    2015-05-01

    Full Text Available From an educational psychology perspective, family life - as a child's primary educational situation - is changing drastically as divorces increase worldwide. Various challenges to relationships accompany the restructuring of family systems after divorce. When remarriage occurs, children's shared membership of two family microsystems and the resultant complexity of the mesosystem cause the reconstituted family situation to come to differ radically from that of a nuclear family. The purpose of this article is to extend Bronfenbrenner's bio-ecological model in order to construct a deeper understanding of the relationship challenges in the context of the reconstituted family, specifically noting the importance of effective parenting at mesosystemic level. Data from two separate qualitative studies was interpreted, based on Bronfenbrenner's bio-ecological model, to form an integrated understanding of the complexity and influence of the mesosystem. The findings indicate that sound proximal interactive processes in the primary and secondary family microsystems depend on an effective mesosystem, and hence, on at least a functionally co-operative relationship between the biological parents after a divorce. Since the biological parents primarily control the effectiveness of the mesosystem, Bronfenbrenner's extended bio-ecological model can be fruitfully applied in all professions dealing with the contextual relationship challenges of reconstituted families.

  18. Reconstitution of high affinity α2 adrenergic agonist binding by fusion with a pertussis toxin substrate

    International Nuclear Information System (INIS)

    Kim, M.H.; Neubig, R.R.

    1986-01-01

    High affinity α 2 adrenergic agonist binding is thought to occur via a coupling of the α 2 receptor with N/sub i/, the inhibitory guanyl nucleotide binding protein. Human platelet membranes pretreated at pH 11.5 exhibit a selective inactivation of agonist binding and N/sub i/. To further study the mechanism of agonist binding, alkali treated membranes (ATM) were mixed with membranes pretreated with 10 μM phenoxybenzamine to block α 2 receptors (POB-M). The combined membrane pellet was incubated in 50% polyethylene glycol (PEG) to promote membrane-membrane fusion and assayed for binding to the α 2 agonist [ 3 H]UK 14,304 (UK) and the antagonist [ 3 H] yohimbine. PEG treatment resulted in a 2-4 fold enhancement of UK binding whereas yohimbine binding was unchanged. No enhancement of UK binding was observed in the absence of PEG treatment. The reconstitution was dependent on the addition of POB-M. They found that a 1:1 ratio of POB-M:ATM was optimal. Reconstituted binding was inhibited by GppNHp. Fusion of rat C6 glioma cell membranes, which do not contain α 2 receptors, also enhanced agonist binding to ATM. Fusion of C6 membranes from cells treated with pertussis toxin did not enhance [ 3 H] UK binding. These data show that a pertussis toxin sensitive membrane component, possibly N/sub i/, can reconstitute high affinity α 2 agonist binding

  19. Engraftment and reconstitution of hematopoiesis is dependent on VEGFR2 mediated regeneration of sinusoidal endothelial cells

    Science.gov (United States)

    Hooper, Andrea T.; Butler, Jason M.; Nolan, Daniel J; Kranz, Andrea; Iida, Kaoruko; Kobayashi, Mariko; Kopp, Hans-Georg; Shido, Koji; Petit, Isabelle; Yanger, Kilangsungla; James, Daylon; Witte, Larry; Zhu, Zhenping; Wu, Yan; Pytowski, Bronislaw; Rosenwaks, Zev; Mittal, Vivek; Sato, Thomas N.; Rafii, Shahin

    2011-01-01

    SUMMARY The phenotypic attributes and molecular determinants for the regeneration of bone marrow (BM) sinusoidal endothelial cells (SECs) and their contribution to hematopoiesis are unknown. We show that after myelosuppression VEGFR2 activation promotes reassembly of regressed SECs, reconstituting hematopoietic stem and progenitor cells (HSPCs). VEGFR2 and VEGFR3 expression are restricted to BM vasculature, demarcating a continuous network of VEGFR2+VEGFR3+Sca1− SECs and VEGFR2+VEGFR3−Sca1+ arterioles. While chemotherapy (5FU) and sublethal irradiation (650 rad) induce minor SEC regression, lethal irradiation (950 rad) induces severe regression of SECs requiring BM transplantation (BMT) for regeneration. Conditional deletion of VEGFR2 in adult mice blocks regeneration of SECs in sublethally irradiated animals, preventing hematopoietic reconstitution. Inhibition of VEGFR2 signaling in lethally irradiated wild type mice rescued with BMT severely impairs SEC reconstruction, preventing engraftment and reconstitution of HSPCs. Therefore, activation of VEGFR2 is critical for regeneration of VEGFR3+Sca1− SECs that are essential for engraftment and restoration of HSPCs and hematopoiesis. PMID:19265665

  20. In vitro reconstitution of the Clostridium botulinum type D progenitor toxin.

    Science.gov (United States)

    Kouguchi, Hirokazu; Watanabe, Toshihiro; Sagane, Yoshimasa; Sunagawa, Hiroyuki; Ohyama, Tohru

    2002-01-25

    Clostridium botulinum type D strain 4947 produces two different sizes of progenitor toxins (M and L) as intact forms without proteolytic processing. The M toxin is composed of neurotoxin (NT) and nontoxic-nonhemagglutinin (NTNHA), whereas the L toxin is composed of the M toxin and hemagglutinin (HA) subcomponents (HA-70, HA-17, and HA-33). The HA-70 subcomponent and the HA-33/17 complex were isolated from the L toxin to near homogeneity by chromatography in the presence of denaturing agents. We were able to demonstrate, for the first time, in vitro reconstitution of the L toxin formed by mixing purified M toxin, HA-70, and HA-33/17. The properties of reconstituted and native L toxins are indistinguishable with respect to their gel filtration profiles, native-PAGE profiles, hemagglutination activity, binding activity to erythrocytes, and oral toxicity to mice. M toxin, which contained nicked NTNHA prepared by treatment with trypsin, could no longer be reconstituted to the L toxin with HA subcomponents, whereas the L toxin treated with proteases was not degraded into M toxin and HA subcomponents. We conclude that the M toxin forms first by assembly of NT with NTNHA and is subsequently converted to the L toxin by assembly with HA-70 and HA-33/17.

  1. Reconstituting factor concentrates: Defining Evidence of Coaching Non-Experts (DEVICE) in haemophilia--a prospective randomized feasibility study.

    Science.gov (United States)

    Bidlingmaier, C; Kurnik, K; Hölscher, G; Kappler, M

    2007-09-01

    The introduction of new needleless devices as demanded by the US Department of Labor Occupational Safety and Health Administration (OSHA) has caused problems with the reconstitution of antihaemophilic factor in emergency situations. Our aim therefore was to evaluate the feasibility of a needleless device for reconstitution of antihaemophilic factor for non-haemophilia experts and to define evidence of the need for coaching these physicians via providing two additional photographs illustrating the two key points of the factor reconstitution process. Twenty-eight physicians of a tertiary care university children's hospital were randomized into two groups, either with no further explanation of the reconstitution device or with two additional photographs, showing the two key steps of the procedure. Reconstitution of dummy-factor concentrate was video-taped and evaluated by a blinded helper. Main outcome measure was the successful reconstitution of dummy-factor concentrate and procedure failure respectively. Of the group without explanation of the reconstitution device, only two of 14 physicians were able to reconstitute the dummy-factor concentrate. Of the group receiving two photographs, nine of 14 completed the task successfully (P = 0.0068). The needleless device is not self explaining to non-haemophilia physicians involved in emergency services. Coaching via short to the point instructions as provided by simple visual educational material therefore is crucial to enable these physicians to resolve the expensive emergency drug quickly and correctly. Companies concerned with the production of any devices to dissolve drugs, especially for treatment of rare diseases as haemophilia, therefore should take measures to simplify therapy.

  2. Development of the plant life management technology for RPV steels [ - Current status of surveillance test specimen reconstitution program -

    International Nuclear Information System (INIS)

    Kazunobu, Sakamoto; Eliichiro, Otsuka; Yoshiaki, Oka; Kosei, Taguchi; Michiyoshi, Yamamoto

    2001-01-01

    In order to develop the reconstitution technology to standardize surveillance test specimen reconstitution practices to deal with the extended service life of reactor pressure vessels, the Japan Power Engineering and Inspection Corporation (JAPEIC) has been carried out the project entrusted by the Ministry of International Trade and Industry (MITI). We focus on a correlation between the reduction of absorbed energy and the interaction of the heat affected zone (HAZ) and the plastic zone, to establish applicable reconstitution conditions for Charpy specimens. The relationship between the plastic zone width and the absorbed energy has been obtained by estimating the plastic zone width from the hardness distribution of the Charpy specimens. Impact tests of reconstituted specimens with 10 mm-length insert using the surface activated joining method were performed and the test results were compared to those obtained by the standard specimens. By comparing the length of insert material to the sum of HAZ width and plastic zone width, it is clear that the interaction causes the reduction of the absorbed energy. Hence, the applicable conditions of reconstituted Charpy specimens could be assessed by comparing the insert length to the sum of HAZ width and plastic zone width. Moreover the effects of the possible deviations from the standard shape and size specimens for the reconstituted specimens were studied. (authors)

  3. Rapid Reconstitution Packages (RRPs) implemented by integration of computational fluid dynamics (CFD) and 3D printed microfluidics.

    Science.gov (United States)

    Chi, Albert; Curi, Sebastian; Clayton, Kevin; Luciano, David; Klauber, Kameron; Alexander-Katz, Alfredo; D'hers, Sebastian; Elman, Noel M

    2014-08-01

    Rapid Reconstitution Packages (RRPs) are portable platforms that integrate microfluidics for rapid reconstitution of lyophilized drugs. Rapid reconstitution of lyophilized drugs using standard vials and syringes is an error-prone process. RRPs were designed using computational fluid dynamics (CFD) techniques to optimize fluidic structures for rapid mixing and integrating physical properties of targeted drugs and diluents. Devices were manufactured using stereo lithography 3D printing for micrometer structural precision and rapid prototyping. Tissue plasminogen activator (tPA) was selected as the initial model drug to test the RRPs as it is unstable in solution. tPA is a thrombolytic drug, stored in lyophilized form, required in emergency settings for which rapid reconstitution is of critical importance. RRP performance and drug stability were evaluated by high-performance liquid chromatography (HPLC) to characterize release kinetics. In addition, enzyme-linked immunosorbent assays (ELISAs) were performed to test for drug activity after the RRPs were exposed to various controlled temperature conditions. Experimental results showed that RRPs provided effective reconstitution of tPA that strongly correlated with CFD results. Simulation and experimental results show that release kinetics can be adjusted by tuning the device structural dimensions and diluent drug physical parameters. The design of RRPs can be tailored for a number of applications by taking into account physical parameters of the active pharmaceutical ingredients (APIs), excipients, and diluents. RRPs are portable platforms that can be utilized for reconstitution of emergency drugs in time-critical therapies.

  4. Maintenance of host leukocytes in peripheral immune compartments following lethal irradiation and bone marrow reconstitution: implications for graft versus host disease.

    Science.gov (United States)

    Staley, Elizabeth M; Tanner, Scott M; Daft, Joseph G; Stanus, Andrea L; Martin, Steven M; Lorenz, Robin G

    2013-03-01

    Bone marrow reconstitution is utilized as a tool for disease treatment and as a research technique to elucidate the function of bone marrow derived cells. Clinically successful engraftment is indicated by the development of a functioning immune repertoire. In research, reconstitution is considered successful if >85% of splenic leukocytes are of donor origins. Previous work suggests that splenic reconstitution may not be indicative of reconstitution in the mucosa. We sought to evaluate mucosal reconstitution in animals following a standard bone marrow eradication and reconstitution technique. Bone marrow was harvested from adult B6.SJL donor mice (CD45.1) and injected via either the retro-orbital or intraperitoneal route into lethally irradiated B6 (CD45.2) adult or neonatal recipients respectively. The expression of CD45 by flow cytometry was used to calculate reconstitution with respect to immune compartment and cell type. In reconstituted adult animals 93.2±1.5% of splenic leukocytes expressed the donor CD45.1 antigen thus meeting the standard definition of reconstitution, however only 58.6±13.6% of intestinal lamina propria lymphocytes and 52.4±16.0% of intestinal intraepithelial lymphocytes were of donor origin, confirming splenic reconstitution fails to represent peripheral immune reconstitution. T-cells in the gastrointestinal tract are the most poorly reconstituted, while B-cells appear to be almost universally replaced by donor cells. The inadequate mucosal reconstitution was not corrected by evaluating later time points or by performing the bone marrow transfer during the neonatal period. This demonstration that substantial host T-cells remain in the intestinal mucosa after a "successful" bone marrow transplantation should cause a re-evaluation of data from research bone marrow chimera experiments, as well as the mechanisms for complications after clinical bone marrow transplantation. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Athenian and Shakespearean Tragedies in Oceania: Teaching Dramatic Literatures in Fiji

    Science.gov (United States)

    Anae, Nicole

    2013-01-01

    This paper presents a theorised classroom-based narrative discussing the author's interdisciplinary approach to the teaching of English dramatic literatures--in particular, Sophocles' "Oedipus the King" and Shakespeare's "Macbeth"--to i-Taukei, Indo-Fijian and Pacific Islander tertiary students at a South Pacific university.…

  6. The Effect of Dramatized Instruction on Speaking Ability of Imam Ali University EFL Learners

    Directory of Open Access Journals (Sweden)

    Sadegh Khosronejad

    2013-09-01

    Full Text Available Teaching language as a second or foreign language, undoubtedly, is so demanding and seeking to find methods for facilitating this prominent practice whets the appetite of any practitioner who works in this field. Research shows that using drama in the classroom as a means of teaching helps students learn socially, academically, and developmentally. This study was an attempt to determine the effect of dramatized instruction on the speaking ability of EFL learners of Imam Ali University. Sixty EFL male students at the intermediate level participated in the study. Their age range was 19-22. Two instruments were utilized in this study; pretest, and posttest.  The data were analyzed through t-test. The data analysis indicated that the mean scores of the experimental group students (M = 72.80 were significantly different (3.29>2; df = 58 from the control group students (M = 65.39. In other words, the experimental group outperformed the control group in the posttest significantly. Moreover, the findings indicated that dramatized instruction does have a great effect on the speaking skills. This study supported the idea of effectiveness of dramatized instruction on developing speaking skill and the teachers can help the learners at lower levels promote their speaking skill through dramatized instruction in EFL classes.

  7. Erotic Language as Dramatic Action in Plays by Lyly and Shakespeare

    Science.gov (United States)

    Knoll, Gillian

    2012-01-01

    This study closely examines the language of desire in the dramatic works of John Lyly and William Shakespeare, and argues that contemplative and analytical speeches about desire function as modes of action in their plays. Erotic speeches do more than express desire in a purely descriptive or perlocutionary capacity distinct from the action of the…

  8. The Devil and his Dramatic Function in Calderon’s Autos Sacramentales

    Directory of Open Access Journals (Sweden)

    Luis Iglesias Feijoo

    2014-05-01

    Full Text Available In some Calderón’s autos sacramentales the character of the Devil draws the plot and establishes the dramatic action as a demiurge. He elaborates plans and strategies to confuse the Man, but he always fails. This function approaches the autos to the metatheater. Finally certain parallels with Shakespeare are suggested.

  9. Art-House Cinema, Avant-Garde Film, and Dramatic Modernism

    Science.gov (United States)

    Cardullo, Bert

    2011-01-01

    In this article, the author talks about art-house cinema, avant-garde film, and dramatic modernism. He believes that the most important modes of film practice are art-house cinema and the avant-garde, both of which contrast with the classical Hollywood mode of film practice. While the latter is characterized by its commercial imperative, corporate…

  10. Collaborative College Playwriting and Performance: A Core Course "Trespassing" onto the Dramatic Arts

    Science.gov (United States)

    Bedetti, Gabriella

    2015-01-01

    Arts integration is relevant in the context of the increased demand for creative thinkers in a global economy. However, reaching across disciplinary boundaries is less common in higher education. Arts integration is one way that a literature class can "trespass" onto the dramatic arts. This paper reports on a study of integrating the…

  11. Survival and growth of Enterobacter sakazakii in infant cereal as affected by composition, reconstitution liquid, and storage temperature.

    Science.gov (United States)

    Lin, Li-Chun; Beuchat, Larry R

    2007-06-01

    Invasive infections caused by Enterobacter sakazakii have occurred predominantly in low-birth-weight neonates and infants younger than 2 months of age. However, infections have also occurred in healthy infants up to 8 months of age and in immunocompromised children up to 4 years of age. The ability of E. sakazakii to survive and grow in infant cereals as affected by composition of the cereal, composition of the reconstitution liquid, and temperature is unknown. A study was done to determine the survival and growth characteristics of E. sakazakii initially at populations of 0.005 and 0.52 CFU/ml of infant rice cereal, oatmeal cereal, or rice with mixed fruit cereal reconstituted with water, milk, or apple juice. Reconstituted cereals were stored at 4, 12, 21, and 30 degrees C, and populations were monitored for up to 72 h. Growth did not occur in reconstituted cereals stored at 4 degrees C or in cereals reconstituted with apple juice and stored at 12 degrees C. Populations (> or =1 CFU/ml) were detected in cereals reconstituted with water or milk and stored at 12, 21, and 30 degres C for 24, 8, and 4 h, respectively. The composition of infant cereals did not markedly affect the survival or growth of E. sakazakii in reconstituted cereals. Populations of E. sakazakii in reconstituted cereal decreased with increases in populations of mesophilic aerobic microflora up to 8 to 9 log CFU/ml, which was concurrent with decreases in pH. E. sakazakii, initially at 2.62 log CFU/ml of rice cereal reconstituted with apple juice (pH 4.32), survived at 40C for at least 14 days. The pathogen grew at 21 and 30 degrees C within 2 days and then decreased to undetectable levels (<1 CFU/10 ml) in cereal stored at 21 degrees C for 5 days or 30'C for 4 days. Initially, at 7.32 log CFU/ml, E. sakazakii was detected in rice cereal stored at 4 degrees C for 50 days. It is recommended that reconstituted infant cereals stored at 21 or 30 degrees C be discarded within 4 h after preparation or

  12. Neurotuberculosis immune reconstitution inflammatory syndrome in the setting of HIV infection: A case report and review of literature

    Directory of Open Access Journals (Sweden)

    Deepasree Jaganmohan

    2016-01-01

    Full Text Available Immune reconstitution inflammatory syndrome (IRIS is an exaggerated immune response which can occur with various coinfections in human immunodeficiency virus (HIV infected patients, of which the most commonly implicated in central nervous system (CNS-IRIS are progressive multifocal leukoencephalopathy (PML, cryptococcosis, and tuberculosis (TB. TB-IRIS is a known complication of pulmonary TB or TB lymphadenitis coinfection in HIV infected patients who are on antituberculosis treatment (ATT after the initiation of antiretroviral therapy (ART. However, development of IRIS in extrapulmonary TB such as CNS TB is very rare. Our case is that of an isolated CNS-TB-IRIS, presenting as increase in the size and perilesional edema of the ring enhancing lesions in the brain, which was observed in two sequential magnetic resonance imaging done over a period of 2 months in a retropositive patient who presented with clinical deterioration after commencement of ART. As prompt diagnosis was made and specific management aimed at IRIS was started without delay, the patient improved symptomatically.

  13. Studies on processing, consumer survey and storage stability of a ready-to-reconstitute fish cutlet mix.

    Science.gov (United States)

    Shaviklo, G Reza; Thorkelsson, Gudjon; Sveinsdottir, Kolbrun; Pourreza, Fatemeh

    2013-10-01

    A convenience ready-to-reconstitute cutlet mix containing 30% fish protein powder was developed to improve the nutritional quality of the product. Consumer survey was based on the home use test (HUT) method. The acceptance of the fish cutlet mix (FCM) was studied using a 9-point hedonic scale ranging from 1 (extremely dislike) to 9 (extremely like). Product's characteristics and stability were studied during 6 months of storage at 27 ± 2 °C. The FCM packed in a polyethylene bag and cardboard box was stable during the storage period. There were no changes in colour, moisture gain and water activity, and TBARS values remained low. The FCM was accepted by the consumers in the study (n = 85). The average liking was high (7.5 ± 1.3) and it was influenced by frequency of fish and chicken consumption, educational level and household size. People who ate fish once a week liked the product more than other consumers. Also those with higher educational level and bigger household size. The results in this paper are important information for companies planning to develop ready-to-eat products fortified with fish proteins. The products could be means of increasing fish consumption in countries/areas where there is no tradition of consuming fresh or frozen fish.

  14. Dramatic Strategies Made Clear: The Feminist Politics in Griselda Gambaro's Puesta en claro

    Directory of Open Access Journals (Sweden)

    Sandra Cypess

    1996-01-01

    Full Text Available In this essay I explore the feminist aspects of Puesta en claro , written in 1974, when Griselda Gambaro was not yet considered a writer who paid attention to feminine issues. Yet to the extent that Gambaro always focuses on the constellation of problems relating to power relations, she is including feminine and feminist issues in her text, and continues a tradition many critics relate to Sor Juana Inés de la Cruz, as I shall argue in my conclusion. Seen within the trajectory of Gambaro's dramaturgy, Puesta en claro , as a play from the 1970s is also remarkable in showing a change in the typical pattern of victimization for her characters. Her audience sees that it is possible for the victims to renounce their passivity and rebel. Moreover, she embodies that message in the sign of a blind, apparently defenseless, domesticated, dominated woman. I suggest that in the creation of the blind woman Gambaro comments on the significance of the male gaze, not as an inherent attribute, but as a controlling agent derived from the patriarchal system. She also shows that the private spaces of the domestic sphere to which women are relegated within the patriarchy can be reconstituted as a site of power.

  15. Ultrasound-Stimulated Drug Delivery Using Therapeutic Reconstituted High-Density Lipoprotein Nanoparticles.

    Science.gov (United States)

    Xiong, Fangyuan; Nirupama, Sabnis; Sirsi, Shashank R; Lacko, Andras; Hoyt, Kenneth

    2017-01-01

    The abnormal tumor vasculature and the resulting abnormal microenvironment are major barriers to optimal chemotherapeutic drug delivery. It is well known that ultrasound (US) can increase the permeability of the tumor vessel walls and enhance the accumulation of anticancer agents. Reconstituted high-density lipoproteins (rHDL) nanoparticles (NPs) allow selective delivery of anticancer agents to tumor cells via their overexpressed scavenger receptor type B1 (SR-B1) receptor. The goal of this study is to investigate the potential of noninvasive US therapy to further improve delivery and tumor uptake of the payload from rHDL NPs, preloaded with an infrared dye (IR-780), aimed to establish a surrogate chemotherapeutic model with optical localization. Athymic nude mice were implanted orthotopically with one million breast cancer cells (MDA-MB-231/Luc). Three weeks later, animals were divided into seven groups with comparable mean tumor size: control, low, moderate, and high concentration of rHDL NPs alone groups, as well as these three levels of rHDL NPs plus US therapy groups ( N = 7 to 12 animals per group), where low, moderate and high denote 5, 10, and 50 µg of the IR-780 dye payload per rHDL NP injection, respectively. The US therapy system included a single element focused transducer connected in series with a function generator and power amplifier. A custom 3D printed cone with an acoustically transparent aperture and filled with degassed water allowed delivery of focused US energy to the tumor tissue. US exposure involved a pulsed sequence applied for a duration of 5 min. Each animal in the US therapy groups received a slow bolus co-injection of MB contrast agent and rHDL NPs. Animals were imaged using a whole-body optical system to quantify intratumoral rHDL NP accumulation at baseline and again at 1 min, 30 min, 24 h, and 48 h. At 48 h, all animals were euthanized and tumors were excised for ex vivo analysis. We investigated a noninvasive optical imaging

  16. Energy transfer and clustering of photosynthetic light-harvesting complexes in reconstituted lipid membranes

    International Nuclear Information System (INIS)

    Dewa, Takehisa; Sumino, Ayumi; Watanabe, Natsuko; Noji, Tomoyasu; Nango, Mamoru

    2013-01-01

    Highlights: ► Photosynthetic light-harvesting complexes were reconstituted into lipid membranes. ► Energy transfers between light-harvesting complexes were examined. ► Atomic force microscopy indicated cluster formation of light-harvesting complexes. ► Efficient energy transfer was observed for the clustered complexes in the membranes. - Abstract: In purple photosynthetic bacteria, light-harvesting complex 2 (LH2) and light harvesting/reaction centre core complex (LH1-RC) play the key roles of capturing and transferring light energy and subsequent charge separation. These photosynthetic apparatuses form a supramolecular assembly; however, how the assembly influences the efficiency of energy conversion is not yet clear. We addressed this issue by evaluating the energy transfer in reconstituted photosynthetic protein complexes LH2 and LH1-RC and studying the structures and the membrane environment of the LH2/LH1-RC assemblies, which had been embedded into various lipid bilayers. Thus, LH2 and LH1-RC from Rhodopseudomonas palustris 2.1.6 were reconstituted in phosphatidylglycerol (PG), phosphatidylcholine (PC), and phosphatidylethanolamine (PE)/PG/cardiolipin (CL). Efficient energy transfer from LH2 to LH1-RC was observed in the PC and PE/PG/CL membranes. Atomic force microscopy revealed that LH2 and LH1-RC were heterogeneously distributed to form clusters in the PC and PE/PG/CL membranes. The results indicated that the phospholipid species influenced the cluster formation of LH2 and LH1-RC as well as the energy transfer efficiency

  17. Energy transfer and clustering of photosynthetic light-harvesting complexes in reconstituted lipid membranes

    Energy Technology Data Exchange (ETDEWEB)

    Dewa, Takehisa, E-mail: takedewa@nitech.ac.jp [Department of Frontier Materials, Graduate School of Engineering, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555 (Japan); Japan Science and Technology, PRESTO, 4-1-8 Honcho Kawaguchi, Saitama 332-0012 (Japan); Sumino, Ayumi; Watanabe, Natsuko; Noji, Tomoyasu [Department of Frontier Materials, Graduate School of Engineering, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555 (Japan); Nango, Mamoru, E-mail: nango@nitech.ac.jp [Department of Frontier Materials, Graduate School of Engineering, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555 (Japan)

    2013-06-20

    Highlights: ► Photosynthetic light-harvesting complexes were reconstituted into lipid membranes. ► Energy transfers between light-harvesting complexes were examined. ► Atomic force microscopy indicated cluster formation of light-harvesting complexes. ► Efficient energy transfer was observed for the clustered complexes in the membranes. - Abstract: In purple photosynthetic bacteria, light-harvesting complex 2 (LH2) and light harvesting/reaction centre core complex (LH1-RC) play the key roles of capturing and transferring light energy and subsequent charge separation. These photosynthetic apparatuses form a supramolecular assembly; however, how the assembly influences the efficiency of energy conversion is not yet clear. We addressed this issue by evaluating the energy transfer in reconstituted photosynthetic protein complexes LH2 and LH1-RC and studying the structures and the membrane environment of the LH2/LH1-RC assemblies, which had been embedded into various lipid bilayers. Thus, LH2 and LH1-RC from Rhodopseudomonas palustris 2.1.6 were reconstituted in phosphatidylglycerol (PG), phosphatidylcholine (PC), and phosphatidylethanolamine (PE)/PG/cardiolipin (CL). Efficient energy transfer from LH2 to LH1-RC was observed in the PC and PE/PG/CL membranes. Atomic force microscopy revealed that LH2 and LH1-RC were heterogeneously distributed to form clusters in the PC and PE/PG/CL membranes. The results indicated that the phospholipid species influenced the cluster formation of LH2 and LH1-RC as well as the energy transfer efficiency.

  18. Reconstitution of experimental neurogenic bladder dysfunction using skeletal muscle-derived multipotent stem cells.

    Science.gov (United States)

    Nitta, Masahiro; Tamaki, Tetsuro; Tono, Kayoko; Okada, Yoshinori; Masuda, Maki; Akatsuka, Akira; Hoshi, Akio; Usui, Yukio; Terachi, Toshiro

    2010-05-15

    BACKGROUND.: Postoperative neurogenic bladder dysfunction is a major complication of radical hysterectomy for cervical cancer and is mainly caused by unavoidable damage to the bladder branch of the pelvic plexus (BBPP) associated with colateral blood vessels. Thus, we attempted to reconstitute disrupted BBPP and blood vessels using skeletal muscle-derived multipotent stem cells that show synchronized reconstitution capacity of vascular, muscular, and peripheral nervous systems. METHODS.: Under pentobarbital anesthesia, intravesical pressure by electrical stimulation of BBPP was measured as bladder function. The distal portion of BBPP with blood vessels was then cut unilaterally (experimental neurogenic bladder model). Measurements were performed before, immediately after, and at 4 weeks after transplantation as functional recovery. Stem cells were obtained from the right soleus and gastrocnemius muscles after enzymatic digestion and cell sorting as CD34/45 (Sk-34) and CD34/45 (Sk-DN). Suspended cells were autografted around the damaged region, whereas medium alone and CD45 cells were transplanted as control groups. To determine the morphological contribution of the transplanted cells, stem cells obtained from green fluorescent protein transgenic mouse muscles were transplanted into a nude rat model and were examined by immunohistochemistry and immunoelectron microscopy. RESULTS.: At 4 weeks after surgery, the transplantation group showed significantly higher functional recovery ( approximately 80%) than the two controls ( approximately 28% and 24%). The transplanted cells showed an incorporation into the damaged peripheral nerves and blood vessels after differentiation into Schwann cells, perineurial cells, vascular smooth muscle cells, pericytes, and fibroblasts around the bladder. CONCLUSION.: Transplantation of multipotent Sk-34 and Sk-DN cells is potentially useful for the reconstitution of damaged BBPP.

  19. Functional reconstitution of prostaglandin E receptor from bovine adrenal medulla with guanine nucleotide binding proteins

    International Nuclear Information System (INIS)

    Negishi, M.; Ito, S.; Yokohama, H.; Hayashi, H.; Katada, T.; Ui, M.; Hayaishi, O.

    1988-01-01

    Prostaglandin E 2 (PEG 2 ) was found to bind specifically to a 100,000 x g pellet prepared from bovine adrenal medulla. The PGE receptor was associated with a GTP-binding protein (G-protein) and could be covalently cross-linked with this G-protein by dithiobis(succinimidyl propionate) in the 100,000 x g pellet. In order to characterize the G-protein associated with the PGE receptor and reconstitute these proteins in phospholipid vesicles, the authors purified the G-protein to apparent homogeneity from the 100,000 x g pellet. The G-protein served as a substrate of pertussis toxin but differed in its α subunit from two known pertussis toxin substrate G-proteins (G/sub i/ and G 0 ) purified from bovine brain. The molecular weight of the α subunit was 40,000, which is between those of G/sub i/ and G 0 . The purified protein was also distinguished immunologically from G/sub i/ and G 0 and was referred to as G/sub am/. Reconstitution of the PGE receptor with pure C/sub am/, G/sub i/, or G 0 in phospholipid vesicles resulted in a remarkable restoration of [ 3 H]PGE 2 binding activity in a GTP-dependent manner. The efficiency of these three G-proteins in this capacity was roughly equal. When pertussis toxin- or N-ethylmaleimide-treated G-proteins, instead of the native ones, were reconstituted into vesicles, the restoration of binding activity was no longer observed. These results indicate that the PGE receptor can couple functionally with G/sub am/, G/sub i/, or G 0 in phospholipid vesicles and suggest that G/sub am/ may be involved in signal transduction of the PGE receptor in bovine adrenal medulla

  20. Coupling of g proteins to reconstituted monomers and tetramers of the M2 muscarinic receptor.

    Science.gov (United States)

    Redka, Dar'ya S; Morizumi, Takefumi; Elmslie, Gwendolynne; Paranthaman, Pranavan; Shivnaraine, Rabindra V; Ellis, John; Ernst, Oliver P; Wells, James W

    2014-08-29

    G protein-coupled receptors can be reconstituted as monomers in nanodiscs and as tetramers in liposomes. When reconstituted with G proteins, both forms enable an allosteric interaction between agonists and guanylyl nucleotides. Both forms, therefore, are candidates for the complex that controls signaling at the level of the receptor. To identify the biologically relevant form, reconstituted monomers and tetramers of the purified M2 muscarinic receptor were compared with muscarinic receptors in sarcolemmal membranes for the effect of guanosine 5'-[β,γ-imido]triphosphate (GMP-PNP) on the inhibition of N-[(3)H]methylscopolamine by the agonist oxotremorine-M. With monomers, a stepwise increase in the concentration of GMP-PNP effected a lateral, rightward shift in the semilogarithmic binding profile (i.e. a progressive decrease in the apparent affinity of oxotremorine-M). With tetramers and receptors in sarcolemmal membranes, GMP-PNP effected a vertical, upward shift (i.e. an apparent redistribution of sites from a state of high affinity to one of low affinity with no change in affinity per se). The data were analyzed in terms of a mechanistic scheme based on a ligand-regulated equilibrium between uncoupled and G protein-coupled receptors (the "ternary complex model"). The model predicts a rightward shift in the presence of GMP-PNP and could not account for the effects at tetramers in vesicles or receptors in sarcolemmal membranes. Monomers present a special case of the model in which agonists and guanylyl nucleotides interact within a complex that is both constitutive and stable. The results favor oligomers of the M2 receptor over monomers as the biologically relevant state for coupling to G proteins. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  1. Paradise, pleasure and desire: Edenic delight in some late-medieval dramatic fragments

    OpenAIRE

    James, Sarah

    2017-01-01

    This paper explores the biblical Paradise and its relationship with the concept of delight or pleasure. In the first section it discusses the changing descriptions and interpretations of Paradise, from the biblical text to later medieval works; it goes on to explore the Augustinian and Thomist philosophies of pleasure and delight. Finally it brings together three late-medieval dramatic texts, all of which share an interest in Paradise, and explores the ways in which these texts utilise the co...

  2. Modeling Attitude towards Drug Treament: The Role of Internal Motivation, External Pressure, and Dramatic Relief

    OpenAIRE

    Conner, Bradley T.; Longshore, Douglas; Anglin, M. Douglas

    2008-01-01

    Motivation for change has historically been viewed as the crucial element affecting responsiveness to drug treatment. Various external pressures, such as legal coercion, may engender motivation in an individual previously resistant to change. Dramatic relief may be the change process that is most salient as individuals internalize such external pressures. Results of structural equation modeling on data from 465 drug users (58.9% male; 21.3% Black, 34.2% Hispanic/Latino, and 35.1% White) enter...

  3. Discrimination, developmental science, and the law: addressing dramatic shifts in civil rights jurisprudence.

    Science.gov (United States)

    Levesque, Roger J R

    2014-01-01

    The civil rights movement fostered dramatic shifts in legal responses to discrimination based on race, gender, and a host of other group characteristics. The legal system now evinces yet another dramatic shift, as it moves from considering difference to focusing on neutrality, from efforts that seek to counter subjugation to those that adopt a "color-blind" approach. The shifting approach already has reached laws regulating responses to the group that spurred massive civil rights reform: minority youth. The shift requires a different body of empirical evidence to address it and a new look at equality jurisprudence. This article notes the need to turn to the current understanding of prejudice and discrimination for guidance, and uses, as illustration, developmental science to shed light on the development, manifestation, and alleviation of invidious discrimination. Using that understanding, the analysis details how the legal system can benefit from that research and better address discrimination in light of dramatic changes in law. The article articulates the need to address discrimination by recognizing and enlisting the law's inculcative powers through multiple sites of inculcation, ranging from families, schools, health and justice systems to religious and community groups. The discussion concludes with brief suggestions for reform benefiting from understandings of prejudice and its expression. (c) 2014 APA, all rights reserved.

  4. Functional Reconstitution of a Fungal Natural Product Gene Cluster by Advanced Genome Editing.

    Science.gov (United States)

    Weber, Jakob; Valiante, Vito; Nødvig, Christina S; Mattern, Derek J; Slotkowski, Rebecca A; Mortensen, Uffe H; Brakhage, Axel A

    2017-01-20

    Filamentous fungi produce varieties of natural products even in a strain dependent manner. However, the genetic basis of chemical speciation between strains is still widely unknown. One example is trypacidin, a natural product of the opportunistic human pathogen Aspergillus fumigatus, which is not produced among different isolates. Combining computational analysis with targeted gene editing, we could link a single nucleotide insertion in the polyketide synthase of the trypacidin biosynthetic pathway and reconstitute its production in a nonproducing strain. Thus, we present a CRISPR/Cas9-based tool for advanced molecular genetic studies in filamentous fungi, exploiting selectable markers separated from the edited locus.

  5. Lack of mucosal immune reconstitution during prolonged treatment of acute and early HIV-1 infection.

    Directory of Open Access Journals (Sweden)

    Saurabh Mehandru

    2006-12-01

    Full Text Available During acute and early HIV-1 infection (AEI, up to 60% of CD4(+ T cells in the lamina propria of the lower gastrointestinal (GI tract are lost as early as 2-4 wk after infection. Reconstitution in the peripheral blood during therapy with highly active antiretroviral therapy (HAART is well established. However, the extent of immune reconstitution in the GI tract is unknown.Fifty-four AEI patients and 18 uninfected control participants underwent colonic biopsy. Forty of the 54 AEI patients were followed after initiation of antiretroviral therapy (18 were studied longitudinally with sequential biopsies over a 3-y period after beginning HAART, and 22 were studied cross sectionally after 1-7 y of uninterrupted therapy. Lymphocyte subsets, markers of immune activation and memory in the peripheral blood and GI tract were determined by flow cytometry and immunohistochemistry. In situ hybridization was performed in order to identify persistent HIV-1 RNA expression. Of the patients studied, 70% maintained, on average, a 50%-60% depletion of lamina propria lymphocytes despite 1-7 y of HAART. Lymphocytes expressing CCR5 and both CCR5 and CXCR4 were persistently and preferentially depleted. Levels of immune activation in the memory cell population, CD45RO+ HLA-DR+, returned to levels seen in the uninfected control participants in the peripheral blood, but were elevated in the GI tract of patients with persistent CD4+ T cell depletion despite therapy. Rare HIV-1 RNA-expressing cells were detected by in situ hybridization.Apparently suppressive treatment with HAART during acute and early infection does not lead to complete immune reconstitution in the GI mucosa in the majority of patients studied, despite immune reconstitution in the peripheral blood. Though the mechanism remains obscure, the data suggest that there is either viral or immune-mediated accelerated T cell destruction or, possibly, alterations in T cell homing to the GI tract. Although clinically

  6. Reusable locking tube insertion and removal fixture and method in a reconstitutable fuel assembly

    International Nuclear Information System (INIS)

    Shallenberger, J.M.; Ferlan, S.J.

    1987-01-01

    This patent describes a locking tube insertion and removal fixture for use with a reconstitutable fuel assembly. The process includes a top nozzle with an adapter plate having at least one passageway, at least one guide thimble with an upper end portion, and an attaching structure having a hollow locking tube insertable and removable into and from a locking position in the upper end portion of the guide thimble for releasably locking the upper end portion of the guide thimble within the passageway of the top nozzle and adapter plate

  7. Mutual Interference between Cytomegalovirus and Reconstitution of Protective Immunity after Hematopoietic Cell Transplantation

    Directory of Open Access Journals (Sweden)

    Matthias J. Reddehase

    2016-08-01

    Full Text Available Hematopoietic cell transplantation (HCT is a therapy option for aggressive forms of hematopoietic malignancies that are resistant to standard antitumoral therapies. Hematoablative treatment preceding HCT, however, opens a ‘window of opportunity’ for latent cytomegalovirus (CMV by releasing it from immune control with the consequence of reactivation of productive viral gene expression and recurrence of infectious virus. A ‘window of opportunity’ for the virus represents a ‘window of risk’ for the patient. In the interim between HCT and reconstitution of antiviral immunity, primarily mediated by CD8+ T cells, initially low amounts of reactivated virus can expand exponentially, disseminate to essentially all organs, and cause multiple organ CMV disease, with interstitial pneumonia (CMV-IP representing the most severe clinical manifestation. Here I will review predictions originally made in the mouse model of experimental HCT and murine CMV infection, some of which have already paved the way to translational preclinical research and promising clinical trials of a pre-emptive cytoimmunotherapy of human CMV disease. Specifically, the mouse model has been pivotal in providing ‘proof of concept’ for preventing CMV disease after HCT by adoptive transfer of preselected, virus epitope-specific effector and memory CD8+ T cells bridging the critical interim. CMV, however, is not a ‘passive antigen’ but is a pathogen that actively interferes with the reconstitution of protective immunity by infecting bone marrow stromal cells that otherwise form niches for hematopoiesis by providing the structural microenvironment and by producing hematopoietically active cytokines, the hemopoietins. Depending on the precise conditions of HCT, reduced homing of transplanted hematopoietic stem- and progenitor cells to infected bone marrow stroma and impaired colony growth and lineage differentiation can lead to ‘graft failure’. In consequence

  8. Functional and morphological recovery of the T-cell compartment in lethally irradiated and reconstituted mice

    International Nuclear Information System (INIS)

    Kraal, G.; Hilst, B. van der; Boden, D.

    1979-01-01

    The recovery of the T-cell compartment in mice after lethal irradiation and reconstitution was studied using functional and morphological parameters. T-helper cell activity, determined by the direct SRBC-plaque-forming cell (PFC) response, recovered in a similar fashion as T-memory function which was studied by adoptive transfer of carrier-primed cells. Both functions returned to control levels in 2.5 to 3 months. Using immunoperoxidase staining of frozen sections with anti-T cell serum, the morphological recovery of the T-cell dependent areas in the white pulp of the spleen could be studied and compared with the functional recovery. (author)

  9. Reconstitutable control assembly having removable control rods with detachable split upper end plugs

    International Nuclear Information System (INIS)

    Gjertsen, R.K.; Knott, R.P.; Sparrow, J.A.

    1989-01-01

    This patent describes, in a reconstitutable control assembly for use with a nuclear fuel assembly, the control assembly including a spider structure and at least one control rod, an attachment joint for detachable fastening the control rod to the spider structure. The attachment joint comprising: a hollow connecting finger on the spider structure; and an elongated detachable split upper end plug on the control rod having a pair of separate upper and lower plug portions, the upper plug portion having integrally-connected tandemly- arranged upper, middle and lower sections. The lower plug portion having integrally-connected tandemly-arranged upper, middle and lower segments

  10. Metallographic study of reconstitution welding in inserts of 1 cm3

    International Nuclear Information System (INIS)

    Romero C, J.; Garcia R, R.; Fernandez T, F.; Perez R, N.; Rocamontes A, M.

    2007-01-01

    In this article, the welding metallographic study carried out in Charpy test tubes reconstituted with notch in 'V', used in the surveillance programs of the vessel in nucleo electric plants is described. Inserts of 1 cm 3 are used, where the inserts are usually rectangular of minimum 18 millimeters of length. The importance of using inserts of 1 cm 3 is that the mechanical properties can be measured in another direction of the vessel steel, when changing the direction or sense of the notch in 'V' or the face where this notch is made in the insert. (Author)

  11. Native and Reconstituted Plasma Lipoproteins in Nanomedicine: Physicochemical Determinants of Nanoparticle Structure, Stability, and Metabolism.

    Science.gov (United States)

    Pownall, Henry J; Rosales, Corina; Gillard, Baiba K; Ferrari, Mauro

    2016-09-01

    Although many acute and chronic diseases are managed via pharmacological means, challenges remain regarding appropriate drug targeting and maintenance of therapeutic levels within target tissues. Advances in nanotechnology will overcome these challenges through the development of lipidic particles, including liposomes, lipoproteins, and reconstituted high-density lipoproteins (rHDL) that are potential carriers of water-soluble, hydrophobic, and amphiphilic molecules. Herein we summarize the properties of human plasma lipoproteins and rHDL, identify the physicochemical determinants of lipid transfer between phospholipid surfaces, and discuss strategies for increasing the plasma half-life of lipoprotein- and liposome-associated molecules.

  12. Neuroimaging of HIV-associated cryptococcal meningitis: comparison of magnetic resonance imaging findings in patients with and without immune reconstitution.

    Science.gov (United States)

    Katchanov, Juri; Branding, Gordian; Jefferys, Laura; Arastéh, Keikawus; Stocker, Hartmut; Siebert, Eberhard

    2016-02-01

    To determine the frequency, imaging characteristics, neuroanatomical distribution and dynamics of magnetic resonance imaging findings in HIV-associated cryptococcal meningitis in immunocompromised patients we compared patients without antiretroviral therapy with patients undergoing immune reconstitution. Neuroimaging and clinical data of 21 consecutive patients presenting to a German HIV centre in a 10-year period between 2005 and 2014 were reviewed. We identified eight patients with magnetic resonance imaging findings related to cryptococcal disease: five patients without antiretroviral therapy and three patients receiving effective antiretroviral therapy resulting in immune reconstitution. The pattern of magnetic resonance imaging manifestations was different in the two groups. In patients not on antiretroviral therapy, pseudocysts (n = 3) and lacunar ischaemic lesions (n = 2) were detected. Contrast-enhancing focal leptomeningeal and/or parenchymal lesions were found in all patients under immune reconstitution (n = 3). Magnetic resonance imaging lesions suggestive of leptomeningitis or meningoencephalitis were detected in all patients with a recurrence of cryptococcal meningitis under immune reconstitution, which differs from the classical magnetic resonance imaging findings in patients without antiretroviral therapy. In antiretroviral therapy-treated patients with past medical history of cryptococcal meningitis, detection of contrast-enhancing focal meningeal and/or parenchymal lesions should prompt further investigations for a recurrence of cryptococcal meningitis under immune reconstitution. © The Author(s) 2015.

  13. Influence of Fiber Bundle Morphology on the Mechanical and Bonding Properties of Cotton Stalk and Mulberry Branch Reconstituted Square Lumber

    Directory of Open Access Journals (Sweden)

    Jing Zhang

    2016-08-01

    Full Text Available The mechanical properties of natural fiber composites can be strengthened in the longitudinal direction if the fiber is formed in a parallel manner. Reconstituted cotton stalk lumber and mulberry branch lumber were fabricated using hot-press technology, and the effects of fiber morphology on their mechanical and bonding properties were investigated. The fiber bundle size had a great influence on the mechanical and bonding properties of the final products. The maximum specific modulus of rupture (MOR and specific modulus of elasticity (MOE of the reconstituted lumber were obtained for medium-size fiber bundles, and the maximum MOR and MOE of reconstituted cotton stalk lumber was 130.3 MPa·g-1·cm-3 and 12.9 GPa·g-1·cm-3, respectively. The maximum MOR and MOE of the mulberry branch lumber was 147.2 MPa·g-1·cm-3 and 14.7 GPa·g-1·cm-3, respectively. Mechanical interlocking structures in the lumber were observed via fluorescence microscopy, showing that phenol-formaldehyde adhesive had penetrated into several cell layers of the fiber bundle under heating and pressure. The adhesive penetration capacity was stronger when the fiber bundles were smaller in size and density. The reconstituted lumber fabricated from both materials exhibited excellent mechanical performance in the parallel direction. Therefore, reconstituted cotton stalk and mulberry branch lumber are attractive potential materials for the construction industry.

  14. Partial reconstitution of virus-specific memory CD8+ T cells following whole body γ-irradiation

    International Nuclear Information System (INIS)

    Grayson, Jason M.; Laniewski, Nathan G.; Holbrook, Beth C.

    2006-01-01

    CD8 + memory T cells are critical in providing immunity to viral infection. Previous studies documented that antigen-specific CD8 + memory T cells are more resistant to radiation-induced apoptosis than naive T cells. Here, we determined the number and in vivo function of memory CD8 + T cells as immune reconstitution progressed following irradiation. Immediately following irradiation, the number of memory CD8 + T cells declined 80%. As reconstitution progressed, the number of memory cells reached a zenith at 33% of pre-irradiation levels, and was maintained for 120 days post-irradiation. In vitro, memory CD8 + T cells were able to produce cytokines at all times post-irradiation, but when adoptively transferred, they were not able to expand upon rechallenge immediately following irradiation, but regained this ability as reconstitution progressed. When proliferation was examined in vitro, irradiated memory CD8 + T cells were able to respond to mitogenic growth but were unable to divide

  15. Active Gating, Molecular Pumping, and Turnover Determination in Biomimetic Lipidic Cubic Mesophases with Reconstituted Membrane Proteins.

    Science.gov (United States)

    Speziale, Chiara; Zabara, Alexandru Florian; Drummond, Calum John; Mezzenga, Raffaele

    2017-11-28

    Understanding the mechanisms controlling molecular transport in bioinspired materials is a central topic in many branches of nanotechnology. In this work, we show that biomolecules of fundamental importance in biological processes, such as glucose, can be transported in an active, controlled, and selective manner across macroscopic lipidic cubic mesophases, by correctly reconstituting within them their corresponding membrane protein transporters, such as Staphylococcus epidermidis (GlcP Se ). Importantly, by duly exploiting the symporter properties of GlcP Se of coupled glucose/H + transport, the diffusion of glucose can further be tuned by independent physiological stimuli, such as parallel or antiparallel pH gradients, offering an important model to study molecular exchange processes in cellular machinery. We finally show that by measuring the transport properties of the lipidic mesophases with and without the GlcP Se membrane protein reconstituted within, it becomes possible to determine its intrinsic conductance. We generalize these findings to other membrane proteins from the antiporters family, such as the bacterial ClC exchanger from Escherichia coli (EcClC), providing a robust method for evaluating the turnover rate of the membrane proteins in general.

  16. Effect of solvent/detergent-treated pooled plasma on fibrinolysis in reconstituted whole blood.

    Science.gov (United States)

    Saadah, Nicholas H; van der Meer, Pieter F; Brinkman, Herm Jan M; de Korte, Dirk; Bontekoe, Ido J; Korsten, Herbert H; Middelburg, Rutger A; van der Bom, Johanna G; Schipperus, Martin R

    2017-10-01

    Hyperfibrinolysis has been observed in patients heavily transfused with solvent/detergent-treated pooled plasma (S/D plasma). We compared coagulation and fibrinolytic variables in blood containing S/D plasma with blood containing fresh-frozen plasma (FFP), with and without α2-antiplasmin or tranexamic acid (TXA) supplementation. Whole blood samples were reconstituted from red blood cells, platelet (PLT) concentrates, and varying mixtures of FFP and S/D plasma. Hematocrit and PLT count of reconstituted whole blood samples were varied. For a subset of runs, α2-antiplasmin or TXA was added to S/D plasma whole blood samples. Thromboelastography (TEG) analysis was performed to assess 50% clot lysis time (CLT 50% ), maximum amplitude (MA), and initial clotting time (R-time). The change in CLT 50% of whole blood as the plasma compartment transitions from FFP to S/D plasma was -52% (95% confidence interval [CI], -60% to -45%; p plasma in whole blood. α2-Antiplasmin and TXA restored clot lysis time in S/D plasma whole blood. Whole blood with S/D plasma has shorter clot lysis times in vitro compared to whole blood with FFP. α2-Antiplasmin and TXA restore clot lysis time of S/D plasma whole blood to that of FFP whole blood. Clinicians should be aware of the decreased clot lysis time associated with S/D plasma transfusion. © 2017 AABB.

  17. Evolutionary biology and anthropology suggest biome reconstitution as a necessary approach toward dealing with immune disorders.

    Science.gov (United States)

    Parker, William; Ollerton, Jeff

    2013-01-01

    Industrialized society currently faces a wide range of non-infectious, immune-related pandemics. These pandemics include a variety of autoimmune, inflammatory and allergic diseases that are often associated with common environmental triggers and with genetic predisposition, but that do not occur in developing societies. In this review, we briefly present the idea that these pandemics are due to a limited number of evolutionary mismatches, the most damaging being 'biome depletion'. This particular mismatch involves the loss of species from the ecosystem of the human body, the human biome, many of which have traditionally been classified as parasites, although some may actually be commensal or even mutualistic. This view, evolved from the 'hygiene hypothesis', encompasses a broad ecological and evolutionary perspective that considers host-symbiont relations as plastic, changing through ecological space and evolutionary time. Fortunately, this perspective provides a blueprint, termed 'biome reconstitution', for disease treatment and especially for disease prevention. Biome reconstitution includes the controlled and population-wide reintroduction (i.e. domestication) of selected species that have been all but eradicated from the human biome in industrialized society and holds great promise for the elimination of pandemics of allergic, inflammatory and autoimmune diseases.

  18. Laser weld reconstitution of conventional Charpy and Miniaturized Notch Test (MNT) specimens

    International Nuclear Information System (INIS)

    Manahan, M.P.; Williams, J.; Martukanitz, R.P.

    1993-01-01

    As nuclear power plants approach end-of-license (EOL) and consideration is given to license renewal, there is an ever increasing need to expand the amount of data obtainable from the original surveillance specimens. A laser welding technique to reconstitute broken Charpy specimens is being developed to produce both conventional and miniaturized Charpy specimens. This paper reports on early laser welding development efforts and summarizes previous proof-of-principle experiments on a 1/16 scale miniaturized Charpy test. In order to benchmark the laser welding procedure, the laser-reconstituted specimen data have been compared with the original specimen data. In addition, the microstructure after welding has been examined to ensure that the material in the vicinity of the notch is essentially unchanged after the welding process. Data which characterize the thermal transient during welding are obtained by attaching thermocouples to the specimens. Other important considerations include perturbation of the stress field near the notch, dynamic stress waves, and contact of the weld region with the tup. Precise control of welding parameters has been demonstrated, heat-affected zones as small as 0.25 mm can be achieved, and sufficient penetration depth can be obtained to enable welding thick sections (1T or greater) to yield conventional Charpy specimens or fracture toughness specimens and thin sections (∼5 mm) to yield Miniaturized Notch Test (MNT) specimens

  19. Use of osmolytes during solubilization and reconstitution of phosphate: sugar phosphate antiport from bacteria

    International Nuclear Information System (INIS)

    Ambudkar, S.V.; Sonna, L.A.; Maloney, P.C.

    1986-01-01

    Phosphate:2-deoxyglucose 6-phosphate (Pi:2DG6P) antiport was extracted from Streptococcus lactis or Staphylococcus aureus with 1.1% octylglucoside in the presence of 0.37% E. coli lipid and reconstituted by detergent dilution. Because previous work suggested inactivation at an early stage, the authors introduced protein stabilants during solubilization. When 20% glycerol was used, proteoliposomes showed a 20-fold increase in 32 Pi transport. This enhanced recovery required phospholipid plus glycerol, and was found only when both were added together with the detergent. Glycerol protection yielded proteoliposomes in which antiporters retained their normal kinetic properties, and Pi exchange by the streptococcal example gave a maximal rate (200-400 nmol/min per mg protein) and a turnover number (30-50/s) which suggested that inactivation had been avoided. Further study showed that 20% glycerol could be replaced by equally high concentrations of compounds classified as osmolytes polyols (erythritol, xylitol, sorbitol), sugars (glucose, trehalose) and certain amino acids (glycine, proline, but not valine). The authors suggest that osmolytes may be used to fully stabilize chemiosmotic transporters during reconstitution

  20. In vitro reconstitution and biochemical analyses of the Schizosaccharomyces pombe nucleosome

    International Nuclear Information System (INIS)

    Koyama, Masako; Nagakura, Wataru; Tanaka, Hiroki; Kujirai, Tomoya; Chikashige, Yuji; Haraguchi, Tokuko; Hiraoka, Yasushi; Kurumizaka, Hitoshi

    2017-01-01

    Schizosaccharomyces pombe, which has a small genome but shares many physiological functions with higher eukaryotes, is a useful single-cell, model eukaryotic organism. In particular, many features concerning chromatin structure and dynamics, including heterochromatin, centromeres, telomeres, and DNA replication origins, are well conserved between S. pombe and higher eukaryotes. However, the S. pombe nucleosome, the fundamental structural unit of chromatin, has not been reconstituted in vitro. In the present study, we established the method to purify S. pombe histones H2A, H2B, H3, and H4, and successfully reconstituted the S. pombe nucleosome in vitro. Our thermal stability assay and micrococcal nuclease treatment assay revealed that the S. pombe nucleosome is markedly unstable and its DNA ends are quite accessible, as compared to the canonical human nucleosome. These findings are important to understand the mechanisms of epigenetic genomic DNA regulation in fission yeast. - Highlights: • S. pombe histones were purified as recombinant proteins. • The recombinant S. pombe histones efficiently form nucleosomes in vitro. • The S. pombe nucleosome has distinct stability and DNA dynamics.

  1. Reconstitution of β-carotene hydroxylase activity of thermostable CYP175A1 monooxygenase

    International Nuclear Information System (INIS)

    Momoi, Kyoko; Hofmann, Ute; Schmid, Rolf D.; Urlacher, Vlada B.

    2006-01-01

    CYP175A1 is a thermostable P450 Monooxygenase from Thermus thermophilus HB27, demonstrating in vivo activity towards β-carotene. Activity of CYP175A1 was reconstituted in vitro using artificial electron transport proteins. First results were obtained in the mixture with a crude Escherichia coli cell extract at 37 o C. In this system, β-carotene was hydroxylated to β-cryptoxanthin. The result indicated the presence of electron transport enzymes among the E. coli proteins, which are suitable for CYP175A1. However, upon in vitro reconstitution of CYP175A1 activity with purified recombinant flavodoxin and flavodoxin reductase from E. coli, only very low β-cryptoxanthin production was observed. Remarkably, with another artificial electron transport system, putidaredoxin and putidaredoxin reductase from Pseudomonas putida, purified CYP175A1 enzyme hydroxylated β-carotene at 3- and also 3'-positions, resulting in β-cryptoxanthin and zeaxanthin. Under the optimal reaction conditions, the turnover rate of the enzyme reached 0.23 nmol β-cryptoxanthin produced per nmol P450 per min

  2. Functional reconstitution of rhodopsin into tubular lipid bilayers supported by nanoporous media.

    Science.gov (United States)

    Soubias, Olivier; Polozov, Ivan V; Teague, Walter E; Yeliseev, Alexei A; Gawrisch, Klaus

    2006-12-26

    We report on a novel reconstitution method for G-protein-coupled receptors (GPCRs) that yields detergent-free, single, tubular membranes in porous anodic aluminum oxide (AAO) filters at concentrations sufficient for structural studies by solid-state NMR. The tubular membranes line the inner surface of pores that traverse the filters, permitting easy removal of detergents during sample preparation as well as delivery of ligands for functional studies. Reconstitution of bovine rhodopsin into AAO filters did not interfere with rhodopsin function. Photoactivation of rhodopsin in AAO pores, monitored by UV-vis spectrophotometry, was indistinguishable from rhodopsin in unsupported unilamellar liposomes. The rhodopsin in AAO pores is G-protein binding competent as shown by a [35S]GTPgammaS binding assay. The lipid-rhodopsin interaction was investigated by 2H NMR on sn-1- or sn-2-chain perdeuterated 1-stearoyl-2-docosahexaenoyl-sn-glycero-3-phospholine as a matrix lipid. Rhodopsin incorporation increased mosaic spread of bilayer orientations and contributed to spectral density of motions with correlation times in the range of nano- to microseconds, detected as a significant reduction in spin-spin relaxation times. The change in lipid chain order parameters due to interaction with rhodopsin was insignificant.

  3. Immunologic reconstitution during PEG-ADA therapy in an unusual mosaic ADA deficient patient.

    Science.gov (United States)

    Liu, Ping; Santisteban, Ines; Burroughs, Lauri M; Ochs, Hans D; Torgerson, Troy R; Hershfield, Michael S; Rawlings, David J; Scharenberg, Andrew M

    2009-02-01

    We report detailed genetic and immunologic studies in a patient diagnosed with adenosine deaminase (ADA) deficiency and combined immune deficiency at age 5 years. At the time of diagnosis, although all other lymphocyte subsets were depleted, circulating CD8(+) T cells with a terminally differentiated phenotype were abundant and expressed normal ADA activity due to a reversion mutation in a CD8(+) T cell or precursor. Over the first 9 months of replacement therapy with PEG-ADA, the patient steadily accumulated mature naïve CD4(+) and CD8(+) T cells, as well as CD4(+)/FOXP3(+) regulatory T cells, consistent with restoration of a functional cellular immune system. While CD19(+) naïve B cells also accumulated in response to PEG-ADA therapy, a high proportion of these B cells exhibited an immature surface marker phenotype even after 9 months, and immunization with neoantigen bacteriophage varphiX174 demonstrated a markedly subnormal humoral immune response. Our observations in this single patient have important implications for gene therapy of human ADA deficiency, as they indicate that ADA expression within even a large circulating lymphocyte population may not be sufficient to support adequate immune reconstitution. They also suggest that an immature surface marker phenotype of the peripheral B cell compartment may be a useful surrogate marker for incomplete humoral immune reconstitution during enzyme replacement, and possibly other forms of hematopoietic cell therapies.

  4. Effect of Different Peat Size and Pre-Consolidation Pressure of Reconstituted Peat on Effective Undrained Shear Strength Properties

    Science.gov (United States)

    Azhar, ATS; Norhaliza, W.; Ismail, B.; Ezree, AM; Nizam, ZM

    2017-08-01

    Shear strength of the soil is one of the most important parameters in engineering design, especially during the pre- or post-construction periods, since it is mainly used to measure and evaluate the foundation or slope stability of soil. Peat normally known as a soil that has a very low value of shear strength, and in order to determine and understand the shear strength of the peat, it is a difficult task in geotechnical engineering due to several factors such as types of fabrics, the origin of the soil, water content, organic matter and the degree of humification. The aim of this study is to determine the effective undrained shear strength properties of reconstituted peat of different sizes. All the reconstituted peat samples were formed with the size that passed the opening sieve of 0.425 mm (effective undrained shear strength properties for reconstituted peat effective shear strength properties for the reconstituted peat effective undrained shear strength properties result obtained from the tests show that the reconstituted peat pore pressure, Δu, show both of reconstituted peat

  5. Heme orientational disorder in human adult hemoglobin reconstituted with a ring fluorinated heme and its functional consequences

    International Nuclear Information System (INIS)

    Nagao, Satoshi; Hirai, Yueki; Kawano, Shin; Imai, Kiyohiro; Suzuki, Akihiro; Yamamoto, Yasuhiko

    2007-01-01

    A ring fluorinated heme, 13,17-bis(2-carboxylatoethyl)-3,8-diethyl-2-fluoro-7,12, 18-trimethyl-porphyrin-atoiron(III), has been incorporated into human adult hemoglobin (Hb A). The heme orientational disorder in the individual subunits of the protein has been readily characterized using 19 F NMR and the O 2 binding properties of the protein have been evaluated through the oxygen equilibrium analysis. The equilibrated orientations of hemes in α- and β- subunits of the reconstituted protein were found to be almost completely opposite to each other, and hence were largely different from those of the native and the previously reported reconstituted proteins [T. Jue, G.N. La Mar, Heme orientational heterogeneity in deuterohemin-reconstituted horse and human hemoglobin characterized by proton nuclear magnetic resonance spectroscopy, Biochem. Biophys. Res. Commun. 119 (1984) 640-645]. Despite the large difference in the degree of the heme orientational disorder in the subunits of the proteins, the O 2 affinity and the cooperativity of the protein reconstituted with 2-MF were similar to those of the proteins reconstituted with a series of hemes chemically modified at the heme 3- and 8-positions [K. Kawabe, K. Imaizumi, Z. Yoshida, K. Imai, I. Tyuma, Studies on reconstituted myoglobins and hemoglobins II. Role of the heme side chains in the oxygenation of hemoglobin, J. Biochem. 92 (1982) 1713-1722], whose O 2 affinity and cooperativity were higher and lower, respectively, relative to those of native protein. These results indicated that the heme orientational disorder could exert little effect, if any, on the O 2 affinity properties of Hb A. This finding provides new insights into structure-function relationship of Hb A

  6. Dramatic Response of a Case ofRecurrent Basal Cell Carcinoma toSystemic Chemotherapy

    Directory of Open Access Journals (Sweden)

    Mohammad Mohammadianpanah

    2010-10-01

    Full Text Available Basal cell carcinoma (BCC is the most common cancer among humans, and the standard treatment is surgery. Other modalities are reserved as a second line of treatment. Topical chemotherapy may be used in primary BCC. Systemic chemotherapy has no role in the primary treatment of BCC, although it may be efficacious in metastatic cases. We report the case of a patient with persistent recurrent BCC following multiple surgeries and radiotherapy, who achieved a dramatic response with a cisplatinand 5-flourouracil chemotherapy regimen.

  7. Dramatic Tumor Shrinkage of Locally Advanced and Inoperable Adenoid Cystic Carcinoma after Intra-arterial Chemotherapy

    Directory of Open Access Journals (Sweden)

    Fu-Jen Hsueh

    2015-06-01

    Full Text Available Adenoid cystic carcinoma is rare and usually arises in the salivary glands. It grows slowly, but is characterized by easy perineural invasion with local infiltration and distant metastasis. In metastatic setting, the efficacy of intravenous chemotherapy is limited. Herein, we report one male patient who had a advanced, inoperable adenoid cystic carcinoma with lung metastasis, presenting with right buccal unhealed ulcer, pain and poor intake, whose loco-regional tumors responded dramatically after intra-arterial chemotherapy and his symptoms were almost completely relieved. We also make a literature review for treatment of adenoid cystic carcinoma.

  8. A proposal for the reorganization of citizenship education via the implementation of the dramatic metod

    Directory of Open Access Journals (Sweden)

    Gökçen Özbek

    2014-04-01

    Full Text Available The purpose of this paper is to establish the relationship between citizenship education and drama method. In this study, I propose a reorganization of the citizenship and/or democracy education by building in a component of drama method. To this end, citizenship education, as well as its applications in Europe and in Turkey, will be explained in order to provide a framework for the concept. Following this, the dramatic method and the connections between drama and citizenship education will be presented for the purpose of demonstrating the possible impact of drama on civic education.

  9. The expressive potential in a dramatic text: Brecht's A Respectable Wedding

    Directory of Open Access Journals (Sweden)

    Katarina Podbevšek

    2017-12-01

    Full Text Available The article discusses the linguistic shaping of a dramatic text and its influence on the text’s stage speech realisation, using the Slovenian translation of Brecht’s one-act play Malomeščanska svatba as an example. A dramatic text typically has a specific – and also graphically visible – textual and linguistic structure that indicates its speech intention. A linguistic analysis of Brecht’s text reveals a great speech potential, both in the stage directions (especially the stage directions for pauses, silence, spoken execution and in the dialogue (characteristic linguistic elements for spontaneous speech. A short comparison of the text with the stage speech performance shows that the actor used not only prosody (especially pauses to semantically enrich and rhythmically organise the written language, but also linguistic interventions into the lexical and syntactic structure (repetition, addition, omission, etc.. The great speech potential of the text thus stimulated the actor’s speech and interpretive creativity.

  10. Consequences of dramatic reductions in state tobacco control funds: Florida, 1998-2000.

    Science.gov (United States)

    Niederdeppe, J; Farrelly, M C; Hersey, J C; Davis, K C

    2008-06-01

    This study assessed whether dramatic funding reductions to the Florida Tobacco Control Program (FTCP) influenced trends in recall of the Florida "truth" anti-smoking media campaign, anti-industry attitudes and non-smoking intentions among Florida teens. We used an interrupted time series technique to test for differences in the rates of change in Florida "truth" recall, anti-industry beliefs and non-smoking intentions before and after the FTCP budget cuts using the Florida Anti-tobacco Media Evaluation (FAME) survey, a repeated cross-sectional telephone survey of Florida teens. Recall of the Florida "truth" anti-smoking campaign, anti-industry attitudes, and non-smoking intentions increased dramatically between April 1998 and May 1999. Florida "truth" recall declined after FTCP budget cuts in June 1999. Anti-industry beliefs and non-smoking intentions plateaued or began to decline after the budget cuts. The launch of the national "truth" campaign in February 2000 may have offset otherwise deleterious effects of the budget cuts on anti-industry beliefs, but not smoking intentions. Reductions in tobacco control funding have immediate effects on programme exposure and cognitive precursors to smoking initiation. There is a critical need to maintain and enhance funding for state tobacco control programmes to continue nationwide progress in preventing youth from initiating cigarette smoking.

  11. Dramatic effect of levetiracetam in early-onset epileptic encephalopathy due to STXBP1 mutation.

    Science.gov (United States)

    Dilena, Robertino; Striano, Pasquale; Traverso, Monica; Viri, Maurizio; Cristofori, Gloria; Tadini, Laura; Barbieri, Sergio; Romeo, Antonino; Zara, Federico

    2016-01-01

    Syntaxin Binding Protein 1 (STXBP1) mutations determine a central neurotransmission dysfunction through impairment of the synaptic vesicle release, thus causing a spectrum of phenotypes varying from syndromic and non-syndromic epilepsy to intellectual disability of variable degree. Among the antiepileptic drugs, levetiracetam has a unique mechanism of action binding SV2A, a glycoprotein of the synaptic vesicle release machinery. We report a 1-month-old boy manifesting an epileptic encephalopathy with clonic seizures refractory to phenobarbital, pyridoxine and phenytoin that presented a dramatic response to levetiracetam with full epilepsy control and EEG normalization. Genetic analysis identified a novel de novo heterozygous mutation (c.[922A>T]p.[Lys308(∗)]) in the STXBP1 gene that severely affects the protein. The observation of a dramatic efficacy of levetiracetam in a case of STXBP1 epileptic encephalopathy refractory to other antiepileptic drugs and considerations regarding the specific mechanism of action of levetiracetam modulating the same system affected by STXBP1 mutations support the hypothesis that this drug may be able to reverse specifically the disease epileptogenic abnormalities. Further clinical observations and laboratory studies are needed to confirm this hypothesis and eventually lead to consider levetiracetam as the first choice treatment of patients with suspected or confirmed STXBP1-related epilepsies. Copyright © 2015 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.

  12. Dramatics and education: Different directions and approaches to the application of drama/theatre techniques

    Directory of Open Access Journals (Sweden)

    Stamenković Ivana

    2015-01-01

    Full Text Available This paper describes, from the perspective of drama pedagogy, two basic directions in the application of drama and/or theatre techniques in the educational context which mainly differ primarily according to who applies them in practice with children and the young - teachers or professional actors.. Then the most prominent approaches of individual authors are singled out from each agent. Although there are certain similarities among them, we wanted also to show their differences, evident in theoretical, pedagogical and practical orientations. In this context the opinions about the importance of dramatics for education, and about the effects of the use of different approaches, levels and the ways of the application of drama/theatre techniques are singled out, showing at the same time different roles, levels and ways of participating students. This theme is important for promoting dramatics pedagogy, which is not at a high level of development in our country, in spite of the fact that many authors are engaged in theoretical and/or practical work in this area.

  13. A Study of Effect of Dramatic Activities on Improving English Communicative Speaking Skill of Grade 11th Students

    Science.gov (United States)

    Iamsaard, Prisana; Kerdpol, Sakon

    2015-01-01

    This paper aimed to reexamine the current EFL communicative speaking skill in high school level in Thailand due to the coming of the entry to ASEAN at the end of the year 2015. Thai students need to be well prepared for workforce in the future since English is used as the working language in ASEAN. The purposes of this paper were to study the…

  14. MHC class II-associated invariant chain linkage of antigen dramatically improves cell-mediated immunity induced by adenovirus vaccines

    DEFF Research Database (Denmark)

    Holst, Peter Johannes; Mandrup Jensen, Camilla Maria; Orskov, Cathrine

    2008-01-01

    The ideal vaccine induces a potent protective immune response, which should be rapidly induced, long-standing, and of broad specificity. Recombinant adenoviral vectors induce potent Ab and CD8+ T cell responses against transgenic Ags within weeks of administration, and they are among the most...

  15. Physical and chemical stability of reconstituted and diluted dexrazoxane infusion solutions.

    Science.gov (United States)

    Zhang, Yan-Ping; Myers, Alan L; Trinh, Van A; Kawedia, Jitesh D; Kramer, Mark A; Benjamin, Robert S; Tran, Hai T

    2014-02-01

    Dexrazoxane is used clinically to prevent anthracycline-associated cardiotoxicity. Hydrolysis of dexrazoxane prior to reaching the cardiac membranes severely hampers its mode of action; therefore, degradation during the preparation and administration of intravenous dexrazoxane admixtures demands special attention. Moreover, the ongoing national shortage of one dexrazoxane formulation in the United States has forced pharmacies to dispense other commercially available dexrazoxane products. However, the manufacturers' limited stability data restrict the flexibility of dexrazoxane usage in clinical practice. The aims of this study are to determine the physical and chemical stability of reconstituted and diluted solutions of two commercially available dexrazoxane formulations. The stability of two dexrazoxane products, brand and generic name, in reconstituted and intravenous solutions stored at room temperature without light protection in polyvinyl chloride bags was determined. The concentrations of dexrazoxane were measured at predetermined time points up to 24 h using a validated reversed phase high-performance liquid chromatography with ultraviolet detection assay. Brand (B-) and generic (G-) dexrazoxane products, reconstituted in either sterile water or 0.167 M sodium lactate (final concentration of 10 mg/mL), were found stable for at least to 8 h. Infusion solutions of B-dexrazoxane, prepared according to each manufacturer's directions, were stable for at least 24 h and 8 h at 1 mg/mL and 3 mg/mL, respectively. Infusion solutions of G-dexrazoxane, prepared in either 5% dextrose or 0.9% sodium chloride following the manufacturer's guidelines, were also stable for at least 24 h and 8 h at 1 mg/mL and 3 mg/mL, respectively. All tested solutions were found physically stable up to 24 h at room temperature. The stability of dexrazoxane infusion solutions reported herein permits advance preparation of dexrazoxane intravenous admixtures, facilitating

  16. Evaluation of Cellular Phenotypes Implicated in Immunopathogenesis and Monitoring Immune Reconstitution Inflammatory Syndrome in HIV/Leprosy Cases

    Science.gov (United States)

    Giacoia-Gripp, Carmem Beatriz Wagner; Sales, Anna Maria; Nery, José Augusto da Costa; Santos-Oliveira, Joanna Reis; de Oliveira, Ariane Leite; Sarno, Euzenir Nunes; Morgado, Mariza Gonçalves

    2011-01-01

    Background It is now evident that HAART-associated immunological improvement often leads to a variety of new clinical manifestations, collectively termed immune reconstitution inflammatory syndrome, or IRIS. This phenomenon has already been described in cases of HIV coinfection with Mycobacterium leprae, most of them belonging to the tuberculoid spectrum of leprosy disease, as observed in leprosy reversal reaction (RR). However, the events related to the pathogenesis of this association need to be clarified. This study investigated the immunological profile of HIV/leprosy patients, with special attention to the cellular activation status, to better understand the mechanisms related to IRIS/RR immunopathogenesis, identifying any potential biomarkers for IRIS/RR intercurrence. Methods/Principal Findings Eighty-five individuals were assessed in this study: HIV/leprosy and HIV-monoinfected patients, grouped according to HIV-viral load levels, leprosy patients without HIV coinfection, and healthy controls. Phenotypes were evaluated by flow cytometry for T cell subsets and immune differentiation/activation markers. As expected, absolute counts of the CD4+ and CD8+ T cells from the HIV-infected individuals changed in relation to those of the leprosy patients and controls. However, there were no significant differences among the groups, whether in the expression of cellular differentiation phenotypes or cellular activation, as reflected by the expression of CD38 and HLA-DR. Six HIV/leprosy patients identified as IRIS/RR were analyzed during IRIS/RR episodes and after prednisone treatment. These patients presented high cellular activation levels regarding the expression of CD38 in CD8+ cells T during IRIS/RR (median: 77,15%), dropping significantly (pleprosy individuals at risk for IRIS/RR. So, a comparative investigation to leprosy patients at RR should be conducted. PMID:22205964

  17. Observational analyses of dramatic developments of a severe air pollution event in the Beijing area

    Science.gov (United States)

    Li, Ju; Sun, Jielun; Zhou, Mingyu; Cheng, Zhigang; Li, Qingchun; Cao, Xiaoyan; Zhang, Jingjiang

    2018-03-01

    A rapid development of a severe air pollution event in Beijing, China, at the end of November 2015 was investigated with unprecedented observations collected during the field campaign of the Study of Urban Rainfall and Fog/Haze (SURF-15). Different from previous statistical analyses of air pollution events and their correlations with meteorological environmental conditions in the area, the role of turbulent mixing in the pollutant transfer was investigated in detail. The analyses indicate that the major pollution source associated with high particulate matter of diameter 2.5 µm (PM2.5) was from south of Beijing. Before the day of the dramatic PM2.5 increase, the nighttime downslope flow from the mountains to the west and north of Beijing reduced the surface PM2.5 concentration northwest of Beijing. The nighttime surface stable boundary layer (SBL) not only kept the relatively less-polluted air near the surface, it also shielded the rough surface from the pollutant transfer by southwesterly winds above the SBL, leading to the fast transport of pollutants over the Beijing area at night. As the daytime convective turbulent mixing developed in the morning, turbulent mixing transported the elevated polluted air downward even though the weak surface wind was from northeast, leading to the dramatic increase of the surface PM2.5 concentration in the urban area. As a result of both turbulent mixing and advection processes with possible aerosol growth from secondary aerosol formation under the low-wind and high-humidity conditions, the PM2.5 concentration reached over 700 µg m-3 in the Beijing area by the end of the day. Contributions of the two transporting processes to the PM2.5 oscillations prior to this dramatic event were also analyzed. The study demonstrates the important role of large-eddy convective turbulent mixing in vertical transfer of pollutants and the role of the SBL in not only decoupling vertical transport of trace gases and aerosols but also in

  18. Quaternary structure of the lactose transport protein of Streptococcus thermophilus in the detergent-solubilized and membrane-reconstituted state

    NARCIS (Netherlands)

    Friesen, R.H.E.; Poolman, B.; Knol, J.

    2000-01-01

    The quaternary structure of LacS, the lactose transporter of Streptococcus thermophilus, has been determined for the detergent-solubilized and the membrane-reconstituted state of the protein. The quaternary structure of the n-dodecyl-β-D-maltoside-solubilized state was studied using a combination of

  19. Reconstituted high-density lipoprotein infusion modulates fatty acid metabolism in patients with type 2 diabetes mellitus

    DEFF Research Database (Denmark)

    Drew, BG; Carey, AL; Natoli, AK

    2011-01-01

    We recently demonstrated that reconstituted high-density lipoprotein (rHDL) modulates glucose metabolism in humans via both AMP-activated protein kinase (AMPK) in muscle and by increasing plasma insulin. Given the key roles of both AMPK and insulin in fatty acid metabolism, the current study inve...

  20. Bread in the Economy of Qualities: The Creative Reconstitution of the Canada-UK Commodity Chain for Wheat

    Science.gov (United States)

    Magnan, Andre

    2011-01-01

    This article traces the creative reconstitution of the Canada-UK wheat-bread commodity chain since the 1990s. In the mid-1990s, the Canadian Wheat Board (CWB) and a British bakery, Warburtons, pioneered an innovative identity-preserved sourcing relationship that ties contracted prairie wheat growers to consumers of premium bread in the United…

  1. Unmasking cryptococcal meningitis immune reconstitution inflammatory syndrome in pregnancy induced by HIV antiretroviral therapy with postpartum paradoxical exacerbation

    Directory of Open Access Journals (Sweden)

    Reuben Kiggundu

    2014-07-01

    Full Text Available Cryptococcosis is the most common cause of meningitis in Africa due to the high burden of HIV. Immune reconstitution inflammatory syndrome (IRIS is a frequent and deadly complication of cryptococcal meningitis. We report a fatal case of cryptococcal-IRIS in a pregnant woman that began after starting antiretroviral therapy (unmasking IRIS and markedly worsened postpartum after delivery (paradoxical IRIS.

  2. Immune reconstitution and risk of Kaposi sarcoma and non-Hodgkin lymphoma in HIV-infected adults

    NARCIS (Netherlands)

    Jaffe, Harold W.; de Stavola, Bianca L.; Carpenter, Lucy M.; Porter, Kholoud; Cox, David R.; del Amo, Julia; Meyer, Laurence; Bucher, Heiner C.; Chêne, Geneviève; Hamouda, Osamah; Pillay, Deenan; Prins, Maria; Rosinska, Magda; Sabin, Caroline; Touloumi, Giota; Lodi, Sara; Coughlin, Kate; Walker, Sarah; Babiker, Abdel; de Luca, Andrea; Fisher, Martin; Muga, Roberto; Zangerle, Robert; Kelleher, A. D.; Cooper, D. A.; Grey, Pat; Finlayson, Robert; Bloch, Mark; Kelleher, Tony; Ramacciotti, Tim; Gelgor, Linda; Cooper, David; Gill, John; Jørgensen, Louise B.; Tartu, U.; Lutsar, Irja; Dabis, Francois; Thiebaut, Rodolphe; Masquelier, Bernard; Costagliola, Dominique; Guiguet, Marguerite; Vanhems, Philippe; Chaix, Marie-Laure; Ghosn, Jade; Boufassa, Faroudy; Kücherer, Claudia; Bartmeyer, Barbara; Geskus, Ronald; van der Helm, Jannie; Schuitemaker, Hanneke

    2011-01-01

    Given the well documented occurrence of immune reconstitution inflammatory syndrome (IRIS) in HIV-infected patients who recently started combination antiretroviral therapy (cART), we examined whether cART initiation increased the risk of Kaposi sarcoma and non-Hodgkin lymphoma (NHL) using data from

  3. Early determinants of long-term T-cell reconstitution after hematopoietic stem cell transplantation for severe combined immunodeficiency

    NARCIS (Netherlands)

    Borghans, José A.; Bredius, Robbert G.; Hazenberg, Mette D.; Roelofs, Helene; Jol-van der Zijde, Els C.; Heidt, Jeroen; Otto, Sigrid A.; Kuijpers, Taco W.; Fibbe, Willem E.; Vossen, Jaak M.; Miedema, Frank; van Tol, Maarten J.

    2006-01-01

    The immune system of patients with severe combined immunodeficiency (SCID) reconstitutes to a large extent during the first years after hematopoietic stem cell transplantation (HSCT). It was suggested, however, that accelerated loss of thymus output may cause impaired immune function at the long

  4. β-Carotene as a factor in the reconstitution of cyclic phospho rylation in damaged chloroplast membranes

    Directory of Open Access Journals (Sweden)

    Anna Tukendorf

    2014-01-01

    Full Text Available Phenazine methosulphate mediated cyclic phosphorylation suppressed by heptane extraction or galactolipase treatment of spinach chloroplasts is restored by β -carotene, in 100% and 50%, respectively. Xanthophylls are not able to reconstitute this reaction. β-Carotene replaces galactolipids in reactivation of galactolipase treated chloroplasts, indicating a nonspecific effect of lipids in photosystem I dependent reactions.

  5. Reconstitution of chlorophyll a/b light-harvesting complexes: xanthophyll-dependent assembly and energy transfer

    Energy Technology Data Exchange (ETDEWEB)

    Plumley, F.G.; Schmidt, G.W.

    1987-01-01

    A method for in vitro reconstitution of the chlorophyll a/b light-harvesting complex from LiDodSO/sub 4//heat-denatured or acetone-extracted photosynthetic membranes has been developed. Characterization of the minimum components necessary for the functional organization or pigments in these membrane complexes reveals that xanthophylls are essential structural components.

  6. Amperometric cholesterol biosensor based on in situ reconstituted cholesterol oxidase on an immobilized monolayer of flavin adenine dinucleotide cofactor.

    Science.gov (United States)

    Vidal, Juan-C; Espuelas, Javier; Castillo, Juan-R

    2004-10-01

    A new amperometric biosensor for determining cholesterol based on deflavination of the enzyme cholesterol oxidase (ChOx) and subsequent reconstitution of the apo-protein with a complexed flavin adenine dinucleotide (FAD) monolayer is described. The charge transfer mediator pyrroquinoline quinone (PQQ) was covalently bound to a cystamine self-assembled monolayer (SAM) on an Au electrode. Boronic acid (BA) was then bound to PQQ using the carbodiimide procedure, and the BA ligand was complexed to the FAD molecules on which the apo-ChOx was subsequently reconstituted. The effective release of the FAD from the enzyme and the successful reconstitution were verified using molecular fluorescence and cyclic voltammetry. The optimal orientation of FAD toward the PQQ mediator and the distances between FAD and PQQ and between PQQ and electrode enhance the charge transfer, very high sensitivity (about 2,500 nAmM(-1)cm(-2)) being obtained for cholesterol determination. The biosensor is selective toward electroactive interferents (ascorbic acid and uric acid) and was tested in reference serum samples, demonstrating excellent accuracy (relative errors below 3% in all cases). The biosensor activity can be successfully regenerated in a simple process by successive reconstitution with batches of recently prepared apo-ChOx on the same immobilized Au/SAM-PQQ-BA-FAD monolayer (it was tested five times); the lifetime of the biosensor is about 45-60 days.

  7. Apolipoprotein A-I configuration and cell cholesterol efflux activity of discoidal lipoproteins depend on the reconstitution process.

    Science.gov (United States)

    Cuellar, Luz Ángela; Prieto, Eduardo Daniel; Cabaleiro, Laura Virginia; Garda, Horacio Alberto

    2014-01-01

    Discoidal high-density lipoproteins (D-HDL) are critical intermediates in reverse cholesterol transport. Most of the present knowledge of D-HDL is based on studies with reconstituted lipoprotein complexes of apolipoprotein A-I (apoA-I) obtained by cholate dialysis (CD). D-HDL can also be generated by the direct microsolubilization (DM) of phospholipid vesicles at the gel/fluid phase transition temperature, a process mechanistically similar to the "in vivo" apoAI lipidation via ABCA1. We compared the apoA-I configuration in D-HDL reconstituted with dimyristoylphosphatidylcholine by both procedures using fluorescence resonance energy transfer measurements with apoA-I tryptophan mutants and fluorescently labeled cysteine mutants. Results indicate that apoA-I configuration in D-HDL depends on the reconstitution process and are consistent with a "double belt" molecular arrangement with different helix registry. As reported by others, a configuration with juxtaposition of helices 5 of each apoAI monomer (5/5 registry) predominates in D-HDL obtained by CD. However, a configuration with helix 5 of one monomer juxtaposed with helix 2 of the other (5/2 registry) would predominate in D-HDL generated by DM. Moreover, we also show that the kinetics of cholesterol efflux from macrophage cultures depends on the reconstitution process, suggesting that apoAI configuration is important for this HDL function. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Enhanced Reconstitution of Human Erythropoiesis and Thrombopoiesis in an Immunodeficient Mouse Model with KitWv Mutations

    Directory of Open Access Journals (Sweden)

    Ayano Yurino

    2016-09-01

    Full Text Available In human-to-mouse xenograft models, reconstitution of human hematopoiesis is usually B-lymphoid dominant. Here we show that the introduction of homozygous KitWv mutations into C57BL/6.Rag2nullIl2rgnull mice with NOD-Sirpa (BRGS strongly promoted human multi-lineage reconstitution. After xenotransplantation of human CD34+CD38− cord blood cells, these newly generated C57BL/6.Rag2nullIl2rgnullNOD-Sirpa KitWv/Wv (BRGSKWv/Wv mice showed significantly higher levels of human cell chimerism and long-term multi-lineage reconstitution compared with BRGS mice. Strikingly, this mouse displayed a robust reconstitution of human erythropoiesis and thrombopoiesis with terminal maturation in the bone marrow. Furthermore, depletion of host macrophages by clodronate administration resulted in the presence of human erythrocytes and platelets in the circulation. Thus, attenuation of mouse KIT signaling greatly enhances the multi-lineage differentiation of human hematopoietic stem and progenitor cells (HSPCs in mouse bone marrow, presumably by outcompeting mouse HSPCs to occupy suitable microenvironments. The BRGSKWv/Wv mouse model is a useful tool to study human multi-lineage hematopoiesis.

  9. Reconstituted influenza virus envelopes as an efficient carrier system for cellular delivery of small-interfering RNAs

    NARCIS (Netherlands)

    de Jonge, J; Holtrop, M; Wilschut, J; Huckriede, A

    Application of RNA interference for in vivo evaluation of gene function or for therapeutic interventions has been hampered by a lack of suitable delivery methods for small interfering RNA ( siRNA). Here, we present reconstituted viral envelopes (virosomes) derived from influenza virus as suitable

  10. De novo reconstitution of a functional mammalian urinary bladder by tissue engineering.

    Science.gov (United States)

    Oberpenning, F; Meng, J; Yoo, J J; Atala, A

    1999-02-01

    Human organ replacement is limited by a donor shortage, problems with tissue compatibility, and rejection. Creation of an organ with autologous tissue would be advantageous. In this study, transplantable urinary bladder neo-organs were reproducibly created in vitro from urothelial and smooth muscle cells grown in culture from canine native bladder biopsies and seeded onto preformed bladder-shaped polymers. The native bladders were subsequently excised from canine donors and replaced with the tissue-engineered neo-organs. In functional evaluations for up to 11 months, the bladder neo-organs demonstrated a normal capacity to retain urine, normal elastic properties, and histologic architecture. This study demonstrates, for the first time, that successful reconstitution of an autonomous hollow organ is possible using tissue-engineering methods.

  11. Reconstitution of a fungal meroterpenoid biosynthesis reveals the involvement of a novel family of terpene cyclases

    Science.gov (United States)

    Itoh, Takayuki; Tokunaga, Kinya; Matsuda, Yudai; Fujii, Isao; Abe, Ikuro; Ebizuka, Yutaka; Kushiro, Tetsuo

    2010-10-01

    Meroterpenoids are hybrid natural products of both terpenoid and polyketide origin. We identified a biosynthetic gene cluster that is responsible for the production of the meroterpenoid pyripyropene in the fungus Aspergillus fumigatus through reconstituted biosynthesis of up to five steps in a heterologous fungal expression system. The cluster revealed a previously unknown terpene cyclase with an unusual sequence and protein primary structure. The wide occurrence of this sequence in other meroterpenoid and indole-diterpene biosynthetic gene clusters indicates the involvement of these enzymes in the biosynthesis of various terpenoid-bearing metabolites produced by fungi and bacteria. In addition, a novel polyketide synthase that incorporated nicotinyl-CoA as the starter unit and a prenyltransferase, similar to that in ubiquinone biosynthesis, was found to be involved in the pyripyropene biosynthesis. The successful production of a pyripyropene analogue illustrates the catalytic versatility of these enzymes for the production of novel analogues with useful biological activities.

  12. Preferential 5-Methylcytosine Oxidation in the Linker Region of Reconstituted Positioned Nucleosomes by Tet1 Protein.

    Science.gov (United States)

    Kizaki, Seiichiro; Zou, Tingting; Li, Yue; Han, Yong-Woon; Suzuki, Yuki; Harada, Yoshie; Sugiyama, Hiroshi

    2016-11-07

    Tet (ten-eleven translocation) family proteins oxidize 5-methylcytosine (mC) to 5-hydroxymethylcytosine (hmC), 5-formylcytosine (fC), and 5-carboxycytosine (caC), and are suggested to be involved in the active DNA demethylation pathway. In this study, we reconstituted positioned mononucleosomes using CpG-methylated 382 bp DNA containing the Widom 601 sequence and recombinant histone octamer, and subjected the nucleosome to treatment with Tet1 protein. The sites of oxidized methylcytosine were identified by bisulfite sequencing. We found that, for the oxidation reaction, Tet1 protein prefers mCs located in the linker region of the nucleosome compared with those located in the core region. © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Biosynthesis of antimycins with a reconstituted 3-formamidosalicylate pharmacophore in Escherichia coli.

    Science.gov (United States)

    Liu, Joyce; Zhu, Xuejun; Seipke, Ryan F; Zhang, Wenjun

    2015-05-15

    Antimycins are a family of natural products generated from a hybrid nonribosomal peptide synthetase (NRPS)-polyketide synthase (PKS) assembly line. Although they possess an array of useful biological activities, their structural complexity makes chemical synthesis challenging, and their biosynthesis has thus far been dependent on slow-growing source organisms. Here, we reconstituted the biosynthesis of antimycins in Escherichia coli, a versatile host that is robust and easy to manipulate genetically. Along with Streptomyces genetic studies, the heterologous expression of different combinations of ant genes enabled us to systematically confirm the functions of the modification enzymes, AntHIJKL and AntO, in the biosynthesis of the 3-formamidosalicylate pharmacophore of antimycins. Our E. coli-based antimycin production system can not only be used to engineer the increased production of these bioactive compounds, but it also paves the way for the facile generation of novel and diverse antimycin analogues through combinatorial biosynthesis.

  14. Reconstituted NALP1 inflammasome reveals two-step mechanism of caspase-1 activation.

    Science.gov (United States)

    Faustin, Benjamin; Lartigue, Lydia; Bruey, Jean-Marie; Luciano, Frederic; Sergienko, Eduard; Bailly-Maitre, Beatrice; Volkmann, Niels; Hanein, Dorit; Rouiller, Isabelle; Reed, John C

    2007-03-09

    Interleukin (IL)-1beta maturation is accomplished by caspase-1-mediated proteolysis, an essential element of innate immunity. NLRs constitute a recently recognized family of caspase-1-activating proteins, which contain a nucleotide-binding oligomerization domain and leucine-rich repeat (LRR) domains and which assemble into multiprotein complexes to create caspase-1-activating platforms called "inflammasomes." Using purified recombinant proteins, we have reconstituted the NALP1 inflammasome and have characterized the requirements for inflammasome assembly and caspase-1 activation. Oligomerization of NALP1 and activation of caspase-1 occur via a two-step mechanism, requiring microbial product, muramyl-dipeptide, a component of peptidoglycan, followed by ribonucleoside triphosphates. Caspase-1 activation by NALP1 does not require but is enhanced by adaptor protein ASC. The findings provide the biochemical basis for understanding how inflammasome assembly and function are regulated, and shed light on NALP1 as a direct sensor of bacterial components in host defense against pathogens.

  15. [Acidity and microbial flora contaminating Senegalese reconstituted curdled milk produced on small scale].

    Science.gov (United States)

    Seydi, M; Ndiaye, M

    1993-01-01

    The present study is carried out to appreciate the acidity, and survey the microbiological quality of reconstituted curdled milk (R.C.M.) obtained on small scale. For his, a total of 100 samples collected from Dakar's sellers were examined. The results show that: pH of R.C.M. is low and very variable: 4.17 +/- 0.30, titrable acidity is high: 152.6 +/- 26.5 degrees D, 5% of samples are contaminated by fecal coliforms and yielded counts of 20 germs/g, 19% of samples are polluted by Staphylococcus aureus to level equal to 10(2) germs/g or higher, yeasts and moulds are present in all samples, salmonella were absent. Saphylococcus aureus survival in curdled milk despite a dysgenesic acidity may be the result of very unhygienic conditions of preparation. Sellers and consumers must be encouraged to respect hygienic rules of preparation. It is the only way to prevent food poisoning.

  16. Cooperative working of bacterial chromosome replication proteins generated by a reconstituted protein expression system

    Science.gov (United States)

    Fujiwara, Kei; Katayama, Tsutomu; Nomura, Shin-ichiro M.

    2013-01-01

    Replication of all living cells relies on the multirounds flow of the central dogma. Especially, expression of DNA replication proteins is a key step to circulate the processes of the central dogma. Here we achieved the entire sequential transcription–translation–replication process by autonomous expression of chromosomal DNA replication machineries from a reconstituted transcription–translation system (PURE system). We found that low temperature is essential to express a complex protein, DNA polymerase III, in a single tube using the PURE system. Addition of the 13 genes, encoding initiator, DNA helicase, helicase loader, RNA primase and DNA polymerase III to the PURE system gave rise to a DNA replication system by a coupling manner. An artificial genetic circuit demonstrated that the DNA produced as a result of the replication is able to provide genetic information for proteins, indicating the in vitro central dogma can sequentially undergo two rounds. PMID:23737447

  17. Bone marrow reconstitution of immune responses following irradiation in the leopard frog, Rana pipiens

    International Nuclear Information System (INIS)

    Ramirez, J.A.; Wright, R.K.; Cooper, E.L.

    1983-01-01

    The bone marrow of Rana is an important source of cells capable of maintaining individual viability, responding to Concanavalin A (Con A) and producing PFC against sheep erythrocyte (SRBC) antigens. Frog marrow is more effective than the spleen in maintaining life. Radiation destroys the ability of frogs to respond to SRBC immunization (lack of bone marrow and spleen PFC, serum antibody) and bone marrow/spleen cells to respond to Con A, i.e., bone marrow and spleen contain radiation-sensitive cells. Shielding one hind leg during irradiation leads to reconstitution of bone marrow/spleen PFC responses, antibody synthesis and individual viability. Our results suggest that bone marrow is: a) the source of stem cells, and b) the source of mature T- and B- lymphocytes that can recirculate within the immune system

  18. Development of reconstitution method for surveillance specimens using surface activated joining

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, Terumi; Kaihara, Shoichiro; Yoshida, Kazuo; Sato, Akira [Ishikawajima-Harima Heavy Industries Co. Ltd., Tokyo (Japan); Onizawa, Kunio; Nishiyama, Yutaka; Fukaya, Kiyoshi; Suzuki, Masahide

    1996-03-01

    Evaluation of embrittlement of reactor vessel steel due to irradiation requires surveillance tests. However, many surveillance specimens are necessary for nuclear plants life extension. Therefore, a specimen reconstitution technique has become important to provide the many specimens for continued surveillance. A surface activated joining (SAJ) method has been developed to join various materials together at low temperatures with little deformation, and is useful to bond irradiated specimens. To assess the validity of this method, Charpy impact tests were carried out, and the characteristics caused by heating during joining were measured. The test results showed the Charpy impact values were almost the same as base materials, and surface activated joining reduced heat affected zone to less than 2 mm. (author).

  19. Specific allogeneic unresponsiveness in irradiated dogs reconstituted with autologous bone marrow

    International Nuclear Information System (INIS)

    Rapaport, F.T.; Bachvaroff, R.J.; Akiyama, N.; Sato, T.; Ferrebee, J.W.

    1980-01-01

    Hemopoietic reconstitution of supralethally irradiated adult dogs of the Cooperstown colony with their own stored bone marrow can produce long-term unresponsiveness to DLA-identical kidney allografts with no need for any additional immunosuppression. Eleven of 18 kidneys transplanted 12 h after replacement of autologous marrow into irradiated recipients currently survive with normal function for as long as 1417 d; 8 of 13 organs transplanted 28 h after marrow replacement, and 8 of 13 organs transplanted 36 h after marrow injection, currently survive up to 502 d, with no further treatment. Alterations in the timing and sequence of each procedure decrease the incidence of unresponsiveness. Survival and function of the kidney allografts were not affected by the rejection of successive skin grafts from the kidney donor. Skin grafts from other DLA-identical donors and DLA-incompatible skin grafts were rejected by the same recipients in uniform fashion

  20. Homogeneous antibodies in lethally irradiated and autologous bone marrow reconstituted Rhesus monkeys

    International Nuclear Information System (INIS)

    Berg, P. Van Den; Radl, J.; Loewenberg, B.; Swart, A.C.W.

    1976-01-01

    Ten Rhesus monkeys were lethally irradiated and reconstituted with autologous bone marrow. During the restoration period, the animals were immunized with DNP-Rhesus albumin and IgA1lambda-10S human paraprotein. One or more transient homogenous immunoglobulin components appeared in sera of all experimental monkeys. In four animals, these homogeneous immunoglobulins were shown to be specific antibodies against DNP-Rhesus albumin. They gradually became as heterogeneous as those in control monkeys which were immunized but not irradiated and transplanted. The onset of the specific antibody response after immunization was slightly delayed in the experimental group. On determining the time necessary to reach normalization of the overall immunoglobulin levels and the normal heterogeneity of the immunoglobulin spectrum, it was found to be more than 1 year in most of the animals. (author)

  1. Expression of human adenosine deaminase in mice reconstituted with retrovirus-transduced hematopoietic stem cells

    International Nuclear Information System (INIS)

    Wilson, J.M.; Danos, O.; Grossman, M.; Raulet, D.H.; Mulligan, R.C.

    1990-01-01

    Recombinant retroviruses encoding human adenosine deaminase have been used to infect murine hematopoietic stem cells. In bone marrow transplant recipients reconstituted with the genetically modified cells, human ADA was detected in peripheral blood mononuclear cells of the recipients for at least 6 months after transplantation. In animals analyzed in detail 4 months after transplantation, human ADA and proviral sequences were detected in all hematopoietic lineages; in several cases, human ADA activity exceeded the endogenous activity. These studies demonstrate the feasibility of introducing a functional human ADA gene into hematopoietic stem cells and obtaining expression in multiple hematopoietic lineages long after transplantation. This approach should be helpful in designing effective gene therapies for severe combined immunodeficiency syndromes in humans

  2. Autophagy in the test tube: In vitro reconstitution of aspects of autophagosome biogenesis.

    Science.gov (United States)

    Rao, Yijian; Matscheko, Nena; Wollert, Thomas

    2016-06-01

    Autophagy is a versatile recycling pathway that delivers cytoplasmic contents to lysosomal compartments for degradation. It involves the formation of a cup-shaped membrane that expands to capture cargo. After the cargo has been entirely enclosed, the membrane is sealed to generate a double-membrane-enclosed compartment, termed the autophagosome. Depending on the physiological state of the cell, the cargo is selected either specifically or non-specifically. The process involves a highly conserved set of autophagy-related proteins. Reconstitution of their action on model membranes in vitro has contributed tremendously to our understanding of autophagosome biogenesis. This review will focus on various in vitro techniques that have been employed to decipher the function of the autophagic core machinery. © 2016 Federation of European Biochemical Societies.

  3. Dramatic response to high-dose icotinib in a lung adenocarcinoma patient after erlotinib failure.

    Science.gov (United States)

    Guan, Yin; Zhao, Hong; Meng, Jing; Yan, Xiang; Jiao, ShunChang

    2014-02-01

    Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) retreatment is rarely administered for non-small cell lung cancer (NSCLC) patients who did not respond to previous TKI treatment. A high dose of TKI may overcome resistance to the standard dose of TKI and have different effectiveness toward cancer compared with the standard dose of TKI. This manuscript describes a dramatic and durable response to high-dose icotinib in a NSCLC patient who did not respond to a previous standard dose of erlotinib. The treatment extended the life of the patient for one additional year. A higher dose of icotinib deserves further study not only for patients whose therapy failed with the standard dose of TKI but also for newly diagnosed NSCLC patients with a sensitive mutation. Serial mutation testing during disease development is necessary for analysis and evaluation of EGFR TKI treatment. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  4. Dramatic reduction of sulfur dioxide emission in Northeastern China in the last decade

    Science.gov (United States)

    Yuan, J.

    2017-12-01

    Analysis of spatial and temporal variations of sulfur dioxide concentration in planetary boundary layer were conducted. The data were generated by NASA satellite daily from October of 2004 and were obtained through NASA Giovanni. The global monthly mean spatial distribution of sulfur dioxide showed several hot spots including: several spots on some islands in the Pacific Ocean, several spots in central America, and central Africa. Most of these hot spots of sulfur dioxide are related to known active volcanos. The biggest hot spot of sulfur dioxide were observed in Northeastern China. While high concentration sulfur dioxide was still observed in Northeastern China in 2017. The area averaged concentration of sulfur dioxide declined dramatically since its peak in 2008. This temporal trend indicates that sulfur reduction effort has been effective in the last decade or post 2008 financial crisis recovery lead an industry less sulfur dioxide emission.

  5. DNA fingerprinting on trial: the dramatic early history of a new forensic technique.

    Science.gov (United States)

    Aronson, Jay D

    2005-09-01

    The early history of "DNA fingerprinting" in the UK might have been different were it not for the accounts of two dramatic courtroom trials, made by the participants and the media, in the mid-1980s. But these reports, which misrepresented the importance DNA evidence had in the trials, left a strong impression on the British public and on judges on both sides of the Atlantic. These trials, widely considered to be the first "victories" for DNA fingerprinting, have been frequently cited as proof of the utility and reliability of the technique, in both the UK and beyond. But in reality, it was the threat of DNA evidence being used rather than the integrity or validity of it that resolved these cases. At that time, DNA fingerprinting was still in its infancy, an untried and untested technology.

  6. Dramatic Clinical Response of Relapsed Metastatic Extramammary Paget’s Disease to Trastuzumab Monotherapy

    Directory of Open Access Journals (Sweden)

    S. Wakabayashi

    2012-01-01

    Full Text Available We report the first case of 68-year-old Japanese woman with metastatic HER2-positive extramammary Paget’s disease that showed the validity of trastuzumab monotherapy. We administered trastuzumab at a loading dose of 8 mg/kg i.v., followed by a 6 mg/kg maintenance dose every three weeks according to a protocol for HER2-positive metastatic breast cancers and a near-complete response was achieved after the tenth infusion. The patient experienced a moderate headache and flushing during the first infusion, but had no advanced effects during subsequent infusions with ibuprofen and d-chlorpheniramine maleate. Given the dramatic response, the patient has had 17 infusions of trastuzumab with no disease progression. Thus, trastuzumab has few side effects and is well tolerated for elderly patients. It may become a new choice of the adjubant therapy of this disease.

  7. Jamie Beck’s “Cinemagraphs” in the context of dramatic theory of literature

    Directory of Open Access Journals (Sweden)

    Ewelina Woźniak

    2012-01-01

    Full Text Available The purpose of this article is to present works of a New York photographer Jamie Beck, who developed a new type of moving photography – cinemagraphs, and look at her work from the perspective of the dramatic theory of literature. Jamie Beck in her works redefines many notions related to the visual realm. Movement and time relationship, the model identity and the impact of material on the image perception are the most interesting problems, that are connected with cinemagraphs. All these issues are analyzed in relation to fields dealing with image: André Rouille’s and Roland Barthes’ photographic theories, art history represented by Georges Didi- Huberman and Rudolf Arnheim’s “the psychology of creative eye”.

  8. Delayed replantation of rat teeth after use of reconstituted powdered milk as a storage medium.

    Science.gov (United States)

    dos Santos, Cláudia Letícia Vendrame; Sonoda, Celso Koogi; Poi, Wilson Roberto; Panzarini, Sônia Regina; Sundefeld, Maria Lúcia Marçal Mazza; Negri, Márcia Regina

    2009-02-01

    Minimal extraoral dry storage period and moist storage for the avulsed tooth are identified as key steps for the treatment protocol of tooth replantation. Among the possible moist storage media, bovine milk has stood out because of its capacity of preserving the integrity of the periodontal ligament (PDL) fibers. This condition has attracted the attention to investigate the use of powdered milk, which is one of the presentation forms of bovine milk, as a feasible storage medium in cases of delayed tooth replantation. The aim of this study was to evaluate the healing process after delayed replantation of rat teeth stored in reconstituted powdered milk and long shelf-life (ultra high temperature) whole milk. Forty maxillary right rat incisors were assigned to four groups (n = 10): group I--the teeth were extracted and immediately replanted into theirs sockets; group II--the teeth were stored for 60 min in 200 ml of freshly reconstituted powdered milk; group III--the teeth were stored for 60 min in 200 ml of long shelf-life whole milk; group IV--the teeth were kept dry for the same time. All procedures were performed at room temperature. Next, the root canals of teeth in groups II, III, and IV were instrumented, filled with a calcium hydroxide-based paste, and replanted into their sockets. All animals received systemic antibiotic therapy and were killed by anesthetic overdose 60 days after replantation. The pieces containing the replanted teeth were removed, fixed, decalcified, and paraffin-embedded. Semi-serial 6-microm-thick sections were obtained and stained with hematoxylin and eosin for histomorphological analysis. There was statistically significant difference (P < 0.05) between groups I and IV regarding the presence of replacement resorption and PDL remnants on root surface. The powdered milk and long shelf-life whole milk presented similar results to each other and may be indicated as storage media for avulsed teeth.

  9. Secreted aspartic proteases are not required for invasion of reconstituted human epithelia by Candida albicans.

    Science.gov (United States)

    Lermann, Ulrich; Morschhäuser, Joachim

    2008-11-01

    A well-known virulence attribute of the human-pathogenic yeast Candida albicans is the secretion of aspartic proteases (Saps), which may contribute to colonization and infection of different host niches by degrading tissue barriers, destroying host defence molecules, or digesting proteins for nutrient supply. The role of individual Sap isoenzymes, which are encoded by a large gene family, for the pathogenicity of C. albicans has been investigated by assessing the virulence of mutants lacking specific SAP genes and by studying the expression pattern of the SAP genes in various models of superficial and systemic infections. We used a recombination-based genetic reporter system to detect the induction of the SAP1-SAP6 genes during infection of reconstituted human vaginal epithelium. Only SAP5, but none of the other tested SAP genes, was detectably activated in this in vitro infection model. To directly address the importance of the SAP1-SAP6 genes for invasion of reconstituted human epithelia (RHE), we constructed a set of mutants of the wild-type C. albicans model strain SC5314 in which either single or multiple SAP genes were specifically deleted. Even mutants lacking all of the SAP1-SAP3 or the SAP4-SAP6 genes displayed the same capacity to invade and damage both oral and vaginal RHE as their wild-type parental strain, in contrast to a nonfilamentous efg1Delta mutant that was avirulent under these conditions. We therefore conclude from these results that the secreted aspartic proteases Sap1p-Sap6p are not required for invasion of RHE by C. albicans.

  10. Synthesis and Self-Assembly of Cellulose Microfibrils from Reconstituted Cellulose Synthase1[OPEN

    Science.gov (United States)

    Purushotham, Pallinti; Fang, Chao; Maranas, Cassandra; Bulone, Vincent

    2017-01-01

    Cellulose, the major component of plant cell walls, can be converted to bioethanol and is thus highly studied. In plants, cellulose is produced by cellulose synthase, a processive family-2 glycosyltransferase. In plant cell walls, individual β-1,4-glucan chains polymerized by CesA are assembled into microfibrils that are frequently bundled into macrofibrils. An in vitro system in which cellulose is synthesized and assembled into fibrils would facilitate detailed study of this process. Here, we report the heterologous expression and partial purification of His-tagged CesA5 from Physcomitrella patens. Immunoblot analysis and mass spectrometry confirmed enrichment of PpCesA5. The recombinant protein was functional when reconstituted into liposomes made from yeast total lipid extract. The functional studies included incorporation of radiolabeled Glc, linkage analysis, and imaging of cellulose microfibril formation using transmission electron microscopy. Several microfibrils were observed either inside or on the outer surface of proteoliposomes, and strikingly, several thinner fibrils formed ordered bundles that either covered the surfaces of proteoliposomes or were spawned from liposome surfaces. We also report this arrangement of fibrils made by proteoliposomes bearing CesA8 from hybrid aspen. These observations describe minimal systems of membrane-reconstituted CesAs that polymerize β-1,4-glucan chains that coalesce to form microfibrils and higher-ordered macrofibrils. How these micro- and macrofibrils relate to those found in primary and secondary plant cell walls is uncertain, but their presence enables further study of the mechanisms that govern the formation and assembly of fibrillar cellulosic structures and cell wall composites during or after the polymerization process controlled by CesA proteins. PMID:28768815

  11. Synthesis and Self-Assembly of Cellulose Microfibrils from Reconstituted Cellulose Synthase.

    Science.gov (United States)

    Cho, Sung Hyun; Purushotham, Pallinti; Fang, Chao; Maranas, Cassandra; Díaz-Moreno, Sara M; Bulone, Vincent; Zimmer, Jochen; Kumar, Manish; Nixon, B Tracy

    2017-09-01

    Cellulose, the major component of plant cell walls, can be converted to bioethanol and is thus highly studied. In plants, cellulose is produced by cellulose synthase, a processive family-2 glycosyltransferase. In plant cell walls, individual β-1,4-glucan chains polymerized by CesA are assembled into microfibrils that are frequently bundled into macrofibrils. An in vitro system in which cellulose is synthesized and assembled into fibrils would facilitate detailed study of this process. Here, we report the heterologous expression and partial purification of His-tagged CesA5 from Physcomitrella patens Immunoblot analysis and mass spectrometry confirmed enrichment of PpCesA5. The recombinant protein was functional when reconstituted into liposomes made from yeast total lipid extract. The functional studies included incorporation of radiolabeled Glc, linkage analysis, and imaging of cellulose microfibril formation using transmission electron microscopy. Several microfibrils were observed either inside or on the outer surface of proteoliposomes, and strikingly, several thinner fibrils formed ordered bundles that either covered the surfaces of proteoliposomes or were spawned from liposome surfaces. We also report this arrangement of fibrils made by proteoliposomes bearing CesA8 from hybrid aspen. These observations describe minimal systems of membrane-reconstituted CesAs that polymerize β-1,4-glucan chains that coalesce to form microfibrils and higher-ordered macrofibrils. How these micro- and macrofibrils relate to those found in primary and secondary plant cell walls is uncertain, but their presence enables further study of the mechanisms that govern the formation and assembly of fibrillar cellulosic structures and cell wall composites during or after the polymerization process controlled by CesA proteins. © 2017 American Society of Plant Biologists. All Rights Reserved.

  12. Reconstitution of DNA strand exchange mediated by Rhp51 recombinase and two mediators.

    Directory of Open Access Journals (Sweden)

    Yumiko Kurokawa

    2008-04-01

    Full Text Available In the fission yeast Schizosaccharomyces pombe, genetic evidence suggests that two mediators, Rad22 (the S. pombe Rad52 homolog and the Swi5-Sfr1 complex, participate in a common pathway of Rhp51 (the S. pombe Rad51 homolog-mediated homologous recombination (HR and HR repair. Here, we have demonstrated an in vitro reconstitution of the central step of DNA strand exchange during HR. Our system consists entirely of homogeneously purified proteins, including Rhp51, the two mediators, and replication protein A (RPA, which reflects genetic requirements in vivo. Using this system, we present the first robust biochemical evidence that concerted action of the two mediators directs the loading of Rhp51 onto single-stranded DNA (ssDNA precoated with RPA. Dissection of the reaction reveals that Rad22 overcomes the inhibitory effect of RPA on Rhp51-Swi5-Sfr1-mediated strand exchange. In addition, Rad22 negates the requirement for a strict order of protein addition to the in vitro system. However, despite the presence of Rad22, Swi5-Sfr1 is still essential for strand exchange. Importantly, Rhp51, but neither Rad22 nor the Swi5-Sfr1 mediator, is the factor that displaces RPA from ssDNA. Swi5-Sfr1 stabilizes Rhp51-ssDNA filaments in an ATP-dependent manner, and this stabilization is correlated with activation of Rhp51 for the strand exchange reaction. Rad22 alone cannot activate the Rhp51 presynaptic filament. AMP-PNP, a nonhydrolyzable ATP analog, induces a similar stabilization of Rhp51, but this stabilization is independent of Swi5-Sfr1. However, hydrolysis of ATP is required for processive strand transfer, which results in the formation of a long heteroduplex. Our in vitro reconstitution system has revealed that the two mediators have indispensable, but distinct, roles for mediating Rhp51 loading onto RPA-precoated ssDNA.

  13. Reconstitution of mammary epithelial morphogenesis by murine embryonic stem cells undergoing hematopoietic stem cell differentiation.

    Directory of Open Access Journals (Sweden)

    Shuxian Jiang

    2010-03-01

    Full Text Available Mammary stem cells are maintained within specific microenvironments and recruited throughout lifetime to reconstitute de novo the mammary gland. Mammary stem cells have been isolated through the identification of specific cell surface markers and in vivo transplantation into cleared mammary fat pads. Accumulating evidence showed that during the reformation of mammary stem cell niches by dispersed epithelial cells in the context of the intact epithelium-free mammary stroma, non-mammary epithelial cells may be sequestered and reprogrammed to perform mammary epithelial cell functions and to adopt mammary epithelial characteristics during reconstruction of mammary epithelium in regenerating mammary tissue in vivo.To examine whether other types of progenitor cells are able to contribute to mammary branching morphogenesis, we examined the potential of murine embryonic stem (mES cells, undergoing hematopoietic differentiation, to support mammary reconstitution in vivo. We observed that cells from day 14 embryoid bodies (EBs under hematopoietic differentiation condition, but not supernatants derived from these cells, when transplanted into denuded mammary fat pads, were able to contribute to both the luminal and myoepithelial lineages in branching ductal structures resembling the ductal-alveolar architecture of the mammary tree. No teratomas were observed when these cells were transplanted in vivo.Our data provide evidence for the dominance of the tissue-specific mammary stem cell niche and its role in directing mES cells, undergoing hematopoietic differentiation, to reprogram into mammary epithelial cells and to promote mammary epithelial morphogenesis. These studies should also provide insights into regeneration of damaged mammary gland and the role of the mammary microenvironment in reprogramming cell fate.

  14. Semisynthetic hemoglobin A: Reconstitution of functional tetramer from semisynthetic α-globin

    International Nuclear Information System (INIS)

    Sahni, G.; Cho, Y.J.; Iyer, K.S.; Khan, S.A.; Seetharam, R.; Acharya, A.S.

    1989-01-01

    The optimal conditions for the semisynthesis of α-globin through Staphylococcus aureus V8 protease condensation of a synthetic fragment (α 1-30 ) with the complementary apo fragment (α 31-141 ) in the presence of structure-inducing organic cosolvents and the reconstitution of the functional tetramer from semisynthetic α-globin have been investigated. The protease-catalyzed ligation of the complementary apo fragments α 1-30 and α 31-141 proceeds with very high selectivity at pH 6.0 and 4 degree C in the presence of 1-propanol as the organic cosolvent. A 30% 1-propanol solution was optimal for the semisynthetic reaction, and the synthetic reaction attained an equilibrium (approximately 50%) in 72 h. The synthetic reaction proceeds smoothly over a wide pH range (pH 5-8). Besides, the semisynthetic system is flexible, and it also proceeded well if trifluoroethanol or 2-propanol was used instead of 1-propanol. However, glycerol, a versatile organic cosolvent used in all other proteosynthetic reactions reported in the literature, was not very efficient as an organic cosolvent in the present synthetic reaction. The semisynthetic α-globin prepared with 1-propanol as the organic cosolvent has been reconstituted into HbA. The semisynthetic HbA was then purified by CM-cellulose chromatography. The semisynthetic HbA is indistinguishable from native HbA, in terms of its structural and functional properties. The semisynthetic approach provides the flexibility in protein engineering studies for the incorporation of spectroscopic labels ( 13 C- and/or 15 N-labeled amino acids), noncoded amino acids, or unnatural bond functionalities, which at present is not possible with genetic approaches

  15. Immune Reconstitution Kinetics following Intentionally Induced Mixed Chimerism by Nonmyeloablative Transplantation.

    Directory of Open Access Journals (Sweden)

    Nayoun Kim

    Full Text Available Establishing mixed chimerism is a promising approach for inducing donor-specific transplant tolerance. The establishment and maintenance of mixed chimerism may enable long-term engraftment of organ transplants while minimizing the use of immunosuppressants. Several protocols for inducing mixed chimerism have been reported; however, the exact mechanism underlying the development of immune tolerance remains to be elucidated. Therefore, understanding the kinetics of engraftment during early post-transplant period may provide insight into establishing long-term mixed chimerism and permanent transplant tolerance. In this study, we intentionally induced allogeneic mixed chimerism using a nonmyeloablative regimen by host natural killer (NK cell depletion and T cell-depleted bone marrow (BM grafts in a major histocompatibility complex (MHC-mismatched murine model and analyzed the kinetics of donor (C57BL/6 and recipient (BALB/c engraftment in the weeks following transplantation. Donor BM cells were well engrafted and stabilized without graft-versus-host disease (GVHD as early as one week post-bone marrow transplantation (BMT. Donor-derived thymic T cells were reconstituted four weeks after BMT; however, the emergence of newly developed T cells was more obvious at the periphery as early as two weeks after BMT. Also, the emergence and changes in ratio of recipient- and donor-derived NKT cells and antigen presenting cells (APCs including dendritic cells (DCs and B cells were noted after BMT. Here, we report a longitudinal analysis of the development of donor- and recipient-originated hematopoietic cells in various lymphatic tissues of intentionally induced mixed chimerism mouse model during early post-transplant period. Through the understanding of immune reconstitution at early time points after nonmyeloablative BMT, we suggest guidelines on intentionally inducing durable mixed chimerism.

  16. Excitation energy transfer between Light-harvesting complex II and Photosystem I in reconstituted membranes.

    Science.gov (United States)

    Akhtar, Parveen; Lingvay, Mónika; Kiss, Teréz; Deák, Róbert; Bóta, Attila; Ughy, Bettina; Garab, Győző; Lambrev, Petar H

    2016-04-01

    Light-harvesting complex II (LHCII), the major peripheral antenna of Photosystem II in plants, participates in several concerted mechanisms for regulation of the excitation energy and electron fluxes in thylakoid membranes. In part, these include interaction of LHCII with Photosystem I (PSI) enhancing the latter's absorption cross-section - for example in the well-known state 1 - state 2 transitions or as a long-term acclimation to high light. In this work we examined the capability of LHCII to deliver excitations to PSI in reconstituted membranes in vitro. Proteoliposomes with native plant thylakoid membrane lipids and different stoichiometric ratios of LHCII:PSI were reconstituted and studied by steady-state and time-resolved fluorescence spectroscopy. Fluorescence emission from LHCII was strongly decreased in PSI-LHCII membranes due to trapping of excitations by PSI. Kinetic modelling of the time-resolved fluorescence data revealed the existence of separate pools of LHCII distinguished by the time scale of energy transfer. A strongly coupled pool, equivalent to one LHCII trimer per PSI, transferred excitations to PSI with near-unity efficiency on a time scale of less than 10ps but extra LHCIIs also contributed significantly to the effective antenna size of PSI, which could be increased by up to 47% in membranes containing 3 LHCII trimers per PSI. The results demonstrate a remarkable competence of LHCII to increase the absorption cross-section of PSI, given the opportunity that the two types of complexes interact in the membrane. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Successful immune reconstitution in severe combined immunodeficiency despite Epstein-Barr virus and cytomegalovirus infections.

    Science.gov (United States)

    DeVoe, P W; Buckley, R H; Shirley, L R; Darby, C P; Ward, F E; Mickey, G H; Raab-Traub, N; Vandenbark, G R

    1985-01-01

    Cytomegalovirus (CMV) and Epstein-Barr virus (EBV), frequently found in the acquired immune deficiency syndrome (AIDS), have been suspected of contributing to the latter immunodeficiency. The ability of normal HLA-identical sibling bone marrow to reconstitute an 8-month-old infant with severe combined immunodeficiency infected with these two viral agents is of interest. After presentation with severe mucocutaneous candidiasis, cavitary pulmonary disease, nodular cutaneous lesions, and hepatic abscesses containing acid-fast organisms, immunologic studies revealed lymphopenia, 1-3% T cells, and no lymphocyte responses to mitogens. Prior to transplantation, the infant's blood B lymphocytes grew spontaneously in culture, suggesting they were infected with EBV. Indeed, an appropriate antibody response to EBV was detected at 2 months post-transplantation. At 3 weeks postgrafting, neutropenia and cholestatic jaundice developed without other signs of graft versus host disease. Liver biopsy demonstrated CMV but no EBV by DNA hybridization. There was evidence of T- and B-cell function by 2 weeks postgrafting, including vigorous in vivo and in vitro responses to candida. Although the blood lymphocyte T4:T8 ratio was inverted at 2 weeks, it reverted to normal by 6 weeks post-transplantation. All clinical disease resolved by 8 months and karotyping revealed all T and B lymphocytes to be XX. Thus, despite infections with both CMV and EBV, complete immunologic reconstitution was achieved in this, the most severe of all genetically determined immunodeficiency conditions, arguing against these viruses having a major role in the failure of bone marrow transplantation in AIDS.

  18. Reconstitution of a secondary cell wall in a secondary cell wall-deficient Arabidopsis mutant.

    Science.gov (United States)

    Sakamoto, Shingo; Mitsuda, Nobutaka

    2015-02-01

    The secondary cell wall constitutes a rigid frame of cells in plant tissues where rigidity is required. Deposition of the secondary cell wall in fiber cells contributes to the production of wood in woody plants. The secondary cell wall is assembled through co-operative activities of many enzymes, and their gene expression is precisely regulated by a pyramidal cascade of transcription factors. Deposition of a transmuted secondary cell wall in empty fiber cells by expressing selected gene(s) in this cascade has not been attempted previously. In this proof-of-concept study, we expressed chimeric activators of 24 transcription factors that are preferentially expressed in the stem, in empty fiber cells of the Arabidopsis nst1-1 nst3-1 double mutant, which lacks a secondary cell wall in fiber cells, under the control of the NST3 promoter. The chimeric activators of MYB46, SND2 and ANAC075, as well as NST3, reconstituted a secondary cell wall with different characteristics from those of the wild type in terms of its composition. The transgenic lines expressing the SND2 or ANAC075 chimeric activator showed increased glucose and xylose, and lower lignin content, whereas the transgenic line expressing the MYB46 chimeric activator showed increased mannose content. The expression profile of downstream genes in each transgenic line was also different from that of the wild type. This study proposed a new screening strategy to identify factors of secondary wall formation and also suggested the potential of the artificially reconstituted secondary cell walls as a novel raw material for production of bioethanol and other chemicals. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists.

  19. In silico reconstitution of actin-based symmetry breaking and motility.

    Directory of Open Access Journals (Sweden)

    Mark J Dayel

    2009-09-01

    Full Text Available Eukaryotic cells assemble viscoelastic networks of crosslinked actin filaments to control their shape, mechanical properties, and motility. One important class of actin network is nucleated by the Arp2/3 complex and drives both membrane protrusion at the leading edge of motile cells and intracellular motility of pathogens such as Listeria monocytogenes. These networks can be reconstituted in vitro from purified components to drive the motility of spherical micron-sized beads. An Elastic Gel model has been successful in explaining how these networks break symmetry, but how they produce directed motile force has been less clear. We have combined numerical simulations with in vitro experiments to reconstitute the behavior of these motile actin networks in silico using an Accumulative Particle-Spring (APS model that builds on the Elastic Gel model, and demonstrates simple intuitive mechanisms for both symmetry breaking and sustained motility. The APS model explains observed transitions between smooth and pulsatile motion as well as subtle variations in network architecture caused by differences in geometry and conditions. Our findings also explain sideways symmetry breaking and motility of elongated beads, and show that elastic recoil, though important for symmetry breaking and pulsatile motion, is not necessary for smooth directional motility. The APS model demonstrates how a small number of viscoelastic network parameters and construction rules suffice to recapture the complex behavior of motile actin networks. The fact that the model not only mirrors our in vitro observations, but also makes novel predictions that we confirm by experiment, suggests that the model captures much of the essence of actin-based motility in this system.

  20. A strategy for solubilizing delipidated apolipoprotein with lysophosphatidylcholine and reconstitution with phosphatidylcholine

    International Nuclear Information System (INIS)

    Kawooya, J.K.; Wells, M.A.; Law, J.H.

    1989-01-01

    The apolipoproteins of insect lipophorin were dissociated in guanidinium chloride and isolated by gel permeation chromatography. Over 98% of the total lipid in lipophorin was associated with apolipophorin I (apoLp-I), thus suggesting this apolipoprotein to be the lipid binding component of the particle. ApoLp-I was delipidated with ethanol/ether and solubilized in buffer that contained radioactive lysophosphatidylcholine ([ 3 H]LPC) above the critical micellar concentration. Sonic irradiation of radioactive phosphatidylcholine ([ 14 C]PC) with [ 3 H]LPC-solubilized apoLp-I at a molar ratio of 318 resulted in reconstituted lipophorin I (RLp-I). [ 3 H]LPC was bound to fatty acid free bovine serum albumin and was separated from RLp-I by density gradient ultracentrifugation and gel permeation chromatography. Negatively stained RLp-I particles were quasispherical with an average radius of 55 angstrom, and their overall morphology and secondary structure were similar to those of native hemolymph lipophorin. The RLp-I particle had a ρ = 1.137 g/mL, a M r ∼ 5.2 x 10 5 , and a [ 14 C]PC:apoLp-I molar ratio of 308. From the compositional analysis, molecular size, trypsinization, and lipolysis with phospholipase A 2 , the authors concluded that each RLp-I particle contained one molecule of apoLp-I and a monomolecular layer of [ 14 C]PC. When injected into the hemolymph of adult moths in vivo, RLp-I was loaded with lipid, as judged by a decrease in its density both in the presence and in the absence of adipokinetic hormone. The similarities in morphology and immunology of RLp-I and native lipophorin, together with the ability of RLp-I to load lipid, suggest that reconstituted lipophorins may serve as models to probe lipophorin structure and function

  1. Reconstitution of hormone-responsive detergent-solubilized follicle stimulating hormone receptors into liposomes

    International Nuclear Information System (INIS)

    Grasso, P.; Dattatreyamurty, B.; Reichert, L.E. Jr.

    1988-01-01

    An FSH receptor-enriched fraction that responds to exogenous FSH by activation of adenylate cyclase was prepared by ultrafiltration of sucrose density gradient-purified light membranes derived from bovine calf testes homogenates and solubilized with Triton X-100. To further confirm the functional nature of the detergent-solubilized FSH receptor, the extract was incorporated by lipid hydration into large multilamellar vesicles composed of dioleoyl phosphatidylcholine and cholesterol, 2:1 molar ratio. Receptor incorporation was determined by measurement of specific binding of [125I] human FSH ([125I] hFSH). Substitution of dioleoyl phosphatidylcholine with dipalmitoyl phosphatidylcholine or increasing the cholesterol concentration of the vesicles reduced specific binding of [125I]hFSH. Under conditions favoring optimal incorporation of the receptor, specific binding of [125I]hFSH was time and temperature dependent and saturable when increasing concentrations of radioligand were added to a constant amount of proteoliposomes. Reconstituted proteoliposomes bound 1600 fmol FSH/mg protein with an affinity of 3.54 x 10(9) M-1. Inhibition of [125I] hFSH binding by hFSH was comparable to that seen with the membrane-bound receptor (ED50 = 10 ng). Equilibrium binding studies with [3H]Gpp(NH)p indicated that a single class of high affinity GTP binding sites with an association constant (Ka) of 3.33 x 10(7) m-1 which bound 2.19 fmol [3H]Gpp(NH)p/mg protein had also been incorporated into the proteoliposomes. Addition of FSH induced a 2-fold stimulation of [3H]Gpp(NH)p binding, supporting our earlier studies suggesting that the detergent-solubilized FSH receptor is complexed to the G protein. Of particular significance in the present study was the observation that both NaF and FSH stimulated cAMP production in the reconstituted system

  2. Time from HIV infection to virological suppression: dramatic fall from 2007 to 2016.

    Science.gov (United States)

    Medland, Nicholas A; Nicholson, Suellen; Chow, Eric P F; Read, Timothy R H; Bradshaw, Catriona S; Denham, Ian; Fairley, Christopher K

    2017-11-13

    Time from HIV infection to virological suppression: dramatic fall from 2007 to 2016. We examined the time from HIV infection to virological suppression in MSM who were first diagnosed at Melbourne Sexual Health Centre between 2007 and 2016. Retrospective cohort. Date of infection was imputed from the testing history or serological evidence of recent infection (negative or indeterminate western blot) or baseline CD4 cell count. Date of virological suppression was determined using clinical viral load data. We analysed predictors of diagnosis with serological evidence of recent infection (logistic regression) and time from diagnosis to suppression and from infection to suppression (Cox regression) using demographic, clinical, and behavioral covariates. Between 2007 and 2016, the median time from HIV infection to diagnosis fell from 6.8 to 4.3 months (P = 0.001), from diagnosis to suppression fell from 22.7 to 3.2 months (P < 0.0001), and from infection to suppression fell from 49.0 to 9.6 months (P < 0.0001). Serological evidence of recent infection increased from 15.6 to 34.3% (P < 0.0001) of diagnoses. In the multivariate analyses, age, being recently arrived from a non-English speaking country, history of IDU, other sexually transmitted infections, and sexual risk were not associated with any of these measures. The duration of infectiousness in MSM diagnosed with HIV infection at Melbourne Sexual Health Centre in Victoria has fallen dramatically between 2007 and 2016 and the proportion diagnosed with serological evidence of recent infection has increased. This effect is observed across all population subgroups and marks a positive milestone for the treatment as prevention paradigm.

  3. Stability assessment of lyophilized intravenous immunoglobulin after reconstitution in glass containers and poly(vinyl chloride) bags.

    Science.gov (United States)

    Parti, R; Mankarious, S

    1997-02-01

    Human intravenous immunoglobulin (IGIV) has been in use for the past 20 years. This biological product is commonly provided in liquid or lyophilized dosage form. When the lyophilized product is rehydrated, it is usually administered within 2-3 h from time of complete dissolution. While this practice is advisable whenever possible, occasionally the patient or care-giver may need to delay the infusion. Hence, a study of the stability of lyophilized IGIV after reconstitution with water for injection was conducted. The reconstituted product was stored either in its original glass container or pooled into poly(vinyl chloride) (PVC) bags. The effect of extended storage on the active ingredient (IgG), excipients (glucose, albumin) and extractables [sodium from glass vials, and di-(2-ethyl-hexyl) phthalate and cyclohexanone from PVC bags] was evaluated. The stability of the active ingredient was evaluated by physico-chemical tests (molecularsize distribution, pH, appearance, total protein), monitoring titres of a specific antibody (hepatitis B surface antigen) and an antibody functional test (bacterial opsonization). To evaluate the risk of microbial contamination during reconstitution and pooling procedures, sterility, pyrogen and animal-safety tests were included in the protocol. The potential of IgG polymerizing in solution during storage and subsequent complement activation was evaluated by assaying for non-specific binding of complement (anti-complement activity). Results show that aseptically reconstituted IGIV is stable and remains sterile up to 48 h at 5 degrees C. The reconstituted product was also found to be stable at room temperature (25 degrees C) up to 12 h.

  4. Nicotinamide mononucleotide inhibits post-ischemic NAD(+) degradation and dramatically ameliorates brain damage following global cerebral ischemia.

    Science.gov (United States)

    Park, Ji H; Long, Aaron; Owens, Katrina; Kristian, Tibor

    2016-11-01

    Nicotinamide adenine dinucleotide (NAD(+)) is an essential cofactor for multiple cellular metabolic reactions and has a central role in energy production. Brain ischemia depletes NAD(+) pools leading to bioenergetics failure and cell death. Nicotinamide mononucleotide (NMN) is utilized by the NAD(+) salvage pathway enzyme, nicotinamide adenylyltransferase (Nmnat) to generate NAD(+). Therefore, we examined whether NMN could protect against ischemic brain damage. Mice were subjected to transient forebrain ischemia and treated with NMN or vehicle at the start of reperfusion or 30min after the ischemic insult. At 2, 4, and 24h of recovery, the proteins poly-ADP-ribosylation (PAR), hippocampal NAD(+) levels, and expression levels of NAD(+) salvage pathway enzymes were determined. Furthermore, animal's neurologic outcome and hippocampal CA1 neuronal death was assessed after six days of reperfusion. NMN (62.5mg/kg) dramatically ameliorated the hippocampal CA1 injury and significantly improved the neurological outcome. Additionally, the post-ischemic NMN treatment prevented the increase in PAR formation and NAD(+) catabolism. Since the NMN administration did not affect animal's temperature, blood gases or regional cerebral blood flow during recovery, the protective effect was not a result of altered reperfusion conditions. These data suggest that administration of NMN at a proper dosage has a strong protective effect against ischemic brain injury. Published by Elsevier Inc.

  5. Who gets custody now? Dramatic changes in children's living arrangements after divorce.

    Science.gov (United States)

    Cancian, Maria; Meyer, Daniel R; Brown, Patricia R; Cook, Steven T

    2014-08-01

    This article reexamines the living arrangements of children following their parents' divorce, using Wisconsin Court Records, updating an analysis that showed relatively small but significant increases in shared custody in the late 1980s and early 1990s. These changes have accelerated markedly in the intervening years: between 1988 and 2008, the proportion of mothers granted sole physical custody fell substantially, the proportion of parents sharing custody increased dramatically, and father-sole custody remained relatively stable. We explore changes in the correlates of alternative custody outcomes, showing that some results from the earlier analysis still hold (for example, cases with higher total family income are more likely to have shared custody), but other differences have lessened (shared-custody cases have become less distinctive as they have become more common). Despite the considerable changes in marriage and divorce patterns over this period, we do not find strong evidence that the changes in custody are related to changes in the characteristics of families experiencing a divorce; rather, changes in custody may be the result of changes in social norms and the process by which custody is determined.

  6. STABILIZING CLOUD FEEDBACK DRAMATICALLY EXPANDS THE HABITABLE ZONE OF TIDALLY LOCKED PLANETS

    International Nuclear Information System (INIS)

    Yang Jun; Abbot, Dorian S.; Cowan, Nicolas B.

    2013-01-01

    The habitable zone (HZ) is the circumstellar region where a planet can sustain surface liquid water. Searching for terrestrial planets in the HZ of nearby stars is the stated goal of ongoing and planned extrasolar planet surveys. Previous estimates of the inner edge of the HZ were based on one-dimensional radiative-convective models. The most serious limitation of these models is the inability to predict cloud behavior. Here we use global climate models with sophisticated cloud schemes to show that due to a stabilizing cloud feedback, tidally locked planets can be habitable at twice the stellar flux found by previous studies. This dramatically expands the HZ and roughly doubles the frequency of habitable planets orbiting red dwarf stars. At high stellar flux, strong convection produces thick water clouds near the substellar location that greatly increase the planetary albedo and reduce surface temperatures. Higher insolation produces stronger substellar convection and therefore higher albedo, making this phenomenon a stabilizing climate feedback. Substellar clouds also effectively block outgoing radiation from the surface, reducing or even completely reversing the thermal emission contrast between dayside and nightside. The presence of substellar water clouds and the resulting clement surface conditions will therefore be detectable with the James Webb Space Telescope.

  7. Patterns and potential drivers of dramatic changes in Tibetan lakes, 1972-2010.

    Science.gov (United States)

    Li, Yingkui; Liao, Jingjuan; Guo, Huadong; Liu, Zewen; Shen, Guozhuang

    2014-01-01

    Most glaciers in the Himalayas and the Tibetan Plateau are retreating, and glacier melt has been emphasized as the dominant driver for recent lake expansions on the Tibetan Plateau. By investigating detailed changes in lake extents and levels across the Tibetan Plateau from Landsat/ICESat data, we found a pattern of dramatic lake changes from 1970 to 2010 (especially after 2000) with a southwest-northeast transition from shrinking, to stable, to rapidly expanding. This pattern is in distinct contrast to the spatial characteristics of glacier retreat, suggesting limited influence of glacier melt on lake dynamics. The plateau-wide pattern of lake change is related to precipitation variation and consistent with the pattern of permafrost degradation induced by rising temperature. More than 79% of lakes we observed on the central-northern plateau (with continuous permafrost) are rapidly expanding, even without glacial contributions, while lakes fed by retreating glaciers in southern regions (with isolated permafrost) are relatively stable or shrinking. Our study shows the limited role of glacier melt and highlights the potentially important contribution of permafrost degradation in predicting future water availability in this region, where understanding these processes is of critical importance to drinking water, agriculture, and hydropower supply of densely populated areas in South and East Asia.

  8. DRAMAtic transforms in magic angle spinning recoupling NMR: The Bessel function pathway.

    Science.gov (United States)

    Goodman, Russell; Hancock, Jason; Siemens, Mark; Jarrell, Harold; Siminovitch, David

    2005-07-01

    In magic angle spinning (MAS) NMR recoupling experiments, the extraction of multiple couplings or a coupling distribution from the observed dephasing signals remains a challenging problem. At least for REDOR experiments, the REDOR transform solves this problem, enabling the simultaneous measurement of multiple dipolar couplings. Focusing on the quadrupolar dephasing observed in QUADRAMA experiments as a representative example, we demonstrate that the same analytical form used for the mathematical description of REDOR dephasing also describes the dephasing observed in a wide variety of MAS NMR recoupling experiments. This fact immediately extends REDOR transform techniques to a much broader suite of recoupling experiments than had previously been realized, including those of DRAMA, MELODRAMA and QUADRAMA. As an illustration, we use the DRAMAtic transform to provide the first inversion of a QUADRAMA dephasing signal to extract the quadrupole coupling distribution. Using a complete elliptic integral of the first kind, we further develop a novel expression for the Pake-spun powder patterns of the corresponding recoupled lineshapes. Our methods and results reinforce the central role that Bessel functions can play in simplifying the integrals that define both the dephasing signals in the time domain, and their Fourier transforms in the frequency domain.

  9. Dramatic Increases of Soil Microbial Functional Gene Diversity at the Treeline Ecotone of Changbai Mountain.

    Science.gov (United States)

    Shen, Congcong; Shi, Yu; Ni, Yingying; Deng, Ye; Van Nostrand, Joy D; He, Zhili; Zhou, Jizhong; Chu, Haiyan

    2016-01-01

    The elevational and latitudinal diversity patterns of microbial taxa have attracted great attention in the past decade. Recently, the distribution of functional attributes has been in the spotlight. Here, we report a study profiling soil microbial communities along an elevation gradient (500-2200 m) on Changbai Mountain. Using a comprehensive functional gene microarray (GeoChip 5.0), we found that microbial functional gene richness exhibited a dramatic increase at the treeline ecotone, but the bacterial taxonomic and phylogenetic diversity based on 16S rRNA gene sequencing did not exhibit such a similar trend. However, the β-diversity (compositional dissimilarity among sites) pattern for both bacterial taxa and functional genes was similar, showing significant elevational distance-decay patterns which presented increased dissimilarity with elevation. The bacterial taxonomic diversity/structure was strongly influenced by soil pH, while the functional gene diversity/structure was significantly correlated with soil dissolved organic carbon (DOC). This finding highlights that soil DOC may be a good predictor in determining the elevational distribution of microbial functional genes. The finding of significant shifts in functional gene diversity at the treeline ecotone could also provide valuable information for predicting the responses of microbial functions to climate change.

  10. Dramatic increases of soil microbial functional gene diversity at the treeline ecotone of Changbai Mountain

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    Congcong Shen

    2016-07-01

    Full Text Available The elevational and latitudinal diversity patterns of microbial taxa have attracted great attention in the past decade. Recently, the distribution of functional attributes has been in the spotlight. Here, we report a study profiling soil microbial communities along an elevation gradient (500 to 2200 m on Changbai Mountain. Using a comprehensive functional gene microarray (GeoChip 5.0, we found that microbial functional gene richness exhibited a dramatic increase at the treeline ecotone, but the bacterial taxonomic and phylogenetic diversity based on 16S rRNA gene sequencing did not exhibit such a similar trend. However, the β-diversity (compositional dissimilarity among sites for both bacterial taxa and functional genes was similar, showing significant elevational distance-decay patterns which presented increased dissimilarity with elevation. The bacterial taxonomic diversity/structure was strongly influenced by soil pH, while the functional gene diversity/structure was significantly correlated with soil dissolved organic carbon (DOC. This finding highlights that soil DOC may be a good predictor in determining the elevational distribution of microbial functional genes. The finding of significant shifts in functional gene diversity at the treeline ecotone could also provide valuable information for predicting the responses of microbial functions to climate change.

  11. mTOR inhibition elicits a dramatic response in PI3K-dependent colon cancers.

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    Dustin A Deming

    Full Text Available The phosphatidylinositide-3-kinase (PI3K signaling pathway is critical for multiple cellular functions including metabolism, proliferation, angiogenesis, and apoptosis, and is the most commonly altered pathway in human cancers. Recently, we developed a novel mouse model of colon cancer in which tumors are initiated by a dominant active PI3K (FC PIK3ca. The cancers in these mice are moderately differentiated invasive mucinous adenocarcinomas of the proximal colon that develop by 50 days of age. Interestingly, these cancers form without a benign intermediary or aberrant WNT signaling, indicating a non-canonical mechanism of tumorigenesis. Since these tumors are dependent upon the PI3K pathway, we investigated the potential for tumor response by the targeting of this pathway with rapamycin, an mTOR inhibitor. A cohort of FC PIK3ca mice were treated with rapamycin at a dose of 6 mg/kg/day or placebo for 14 days. FDG dual hybrid PET/CT imaging demonstrated a dramatic tumor response in the rapamycin arm and this was confirmed on necropsy. The tumor tissue remaining after treatment with rapamycin demonstrated increased pERK1/2 or persistent phosphorylated ribosomal protein S6 (pS6, indicating potential resistance mechanisms. This unique model will further our understanding of human disease and facilitate the development of therapeutics through pharmacologic screening and biomarker identification.

  12. Membrane Proteins Are Dramatically Less Conserved than Water-Soluble Proteins across the Tree of Life.

    Science.gov (United States)

    Sojo, Victor; Dessimoz, Christophe; Pomiankowski, Andrew; Lane, Nick

    2016-11-01

    Membrane proteins are crucial in transport, signaling, bioenergetics, catalysis, and as drug targets. Here, we show that membrane proteins have dramatically fewer detectable orthologs than water-soluble proteins, less than half in most species analyzed. This sparse distribution could reflect rapid divergence or gene loss. We find that both mechanisms operate. First, membrane proteins evolve faster than water-soluble proteins, particularly in their exterior-facing portions. Second, we demonstrate that predicted ancestral membrane proteins are preferentially lost compared with water-soluble proteins in closely related species of archaea and bacteria. These patterns are consistent across the whole tree of life, and in each of the three domains of archaea, bacteria, and eukaryotes. Our findings point to a fundamental evolutionary principle: membrane proteins evolve faster due to stronger adaptive selection in changing environments, whereas cytosolic proteins are under more stringent purifying selection in the homeostatic interior of the cell. This effect should be strongest in prokaryotes, weaker in unicellular eukaryotes (with intracellular membranes), and weakest in multicellular eukaryotes (with extracellular homeostasis). We demonstrate that this is indeed the case. Similarly, we show that extracellular water-soluble proteins exhibit an even stronger pattern of low homology than membrane proteins. These striking differences in conservation of membrane proteins versus water-soluble proteins have important implications for evolution and medicine. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  13. Regulatory Bodies: Dramatic Creativity, Control and the Commodity of Lady Audley's Secret

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    Kate Mattacks

    2009-04-01

    Full Text Available 'Lady Audley's Secret' resonates with a sense of theatricality grounded in the eponymous heroine's propensity for visual tactics to attract an audience. Repositioning the dramatic exchange between actress and audience within the domestic sphere sensitised the reader to the question of diagnosing morally degenerative artificiality whilst involving the reader in the complicit act of encouraging Lady Audley's performance through consumer demand. This complex relationship between consumerism, theatre and society saw Lady Audley's figure assume an iconic status, but little has been written on the creative interplay between the novel and the theatre itself. Beginning with the way in which aspiring actresses used Lady Audley as a model to display their histrionic variety, I argue that the theatre used her character to engage with the complex connections between drama and commodity culture. Her autophagy in favour of a false identity provided a model for actresses to visualise self-commodification whilst highlighting the tension between the material practices of the theatre and the materiality of commodity culture. The drive for regulation of the theatrical medium finds its locus in the licensing text/published acting edition of William E. Suter's 'Lady Audley's Secret' (1863. Here I move to show how Suter's adaptation functioned as a site where the state regulation of the theatre was explored and contested. These unauthorised images of Lady Audley's containment came to symbolise not only the regulatory processes of licensing and copyright, but also the creative exploitation of a commodity culture critiqued through the dehumanised form of Braddon's original heroine.

  14. Analysing the development of TV news programmes: from information to dramatization

    Directory of Open Access Journals (Sweden)

    Maribel Rodríguez Fidalgo, Ph.D.

    2010-01-01

    Full Text Available Abstract: The TV news programme is the backbone of all television networks, be they national, regional or local. These types of programmes have been changing over the time. The essence of the information is being modified and has adopted a new format. Based on these principles, this research analyses the evolution of the language used on the Spanish TV news programmes, from their origins until today. This research has been carried out in three different phases: late 1980s, the 1990s and recent years. These stages were thoroughly examined through surveys applied to representative samples of the population. The results showed that TV news programmes have been acquiring a changing narrative style throughout time: they began using an “objective” narrative, followed by a mediated type, and finally dramatized narrative which uses shocking visual content and aims to achieve audience’s sensibilization and identification with news’ protagonists. The spectacularization of information has become the identitary feature of the current TV news programmes as a response to the need of reaching higher audience ratings.

  15. Synthesis and characterization of tethered lipid assemblies for membrane protein reconstitution (Review).

    Science.gov (United States)

    Veneziano, Rémi; Rossi, Claire; Chenal, Alexandre; Brenner, Catherine; Ladant, Daniel; Chopineau, Joël

    2017-09-28

    Biological membranes and their related molecular mechanisms are essential for all living organisms. Membranes host numerous proteins and are responsible for the exchange of molecules and ions, cell signaling, and cell compartmentation. Indeed, the plasma membrane delimits the intracellular compartment from the extracellular environment and intracellular membranes. Biological membranes also play a major role in metabolism regulation and cellular physiology (e.g., mitochondrial membranes). The elaboration of membrane based biomimetic systems allows us to reconstitute and investigate, in controlled conditions, biological events occurring at the membrane interface. A whole variety of model membrane systems have been developed in the last few decades. Among these models, supported membranes were developed on various hydrophilic supports. The use of solid supports enables the direct use of surface sensitive techniques (e.g., surface plasmon resonance, quartz crystal microbalance, and atomic force microscopy) to monitor and quantify events occurring at the membrane surface. Tethered bilayer membranes (tBLMs) could be considered as an achievement of the first solid supported membranes described by the McConnell group. Tethered bilayers on solid supports were designed to delimit an inside compartment from an outside one. They were used for measuring interactions with ligands or incorporating large membrane proteins or complexes without interference with the support. In this context, the authors developed an easy concept of versatile tBLMs assembled on amino coated substrates that are formed upon the vesicle fusion rupture process applicable to protein-free vesicles as well as proteoliposomes. The phospholipid bilayer (natural or synthetic lipids) incorporated 5% of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-poly ethylene glycol-N-hydroxy succinimide to ensure the anchorage of the bilayer to the amino coated surface. The conditions for the formation of tBLMs on amino

  16. Effect of pH adjustment, homogenization and diafiltration on physicochemical, reconstitution, functional and rheological properties of medium protein milk protein concentrates (MPC70).

    Science.gov (United States)

    Meena, Ganga Sahay; Singh, Ashish Kumar; Gupta, Vijay Kumar; Borad, Sanket; Arora, Sumit; Tomar, Sudhir Kumar

    2018-04-01

    Poor solubility is the major limiting factor in commercial applications of milk protein concentrates (MPC) powders. Retentate treatments such as pH adjustment using disodium phosphate (Na 2 HPO 4 ), also responsible for calcium chelation with homogenization and; its diafiltration with 150 mM NaCl solution were hypothesized to improve the functional properties of treated MPC70 powders. These treatments significantly improved the solubility, heat stability, water binding, dispersibility, bulk density, flowability, buffer index, foaming and emulsifying capacity of treated powders over control. Rheological behaviour of reconstituted MPC solutions was best explained by Herschel Bulkley model. Compared to rough, large globular structures with dents in control; majorly intact, separate, smaller particles of smooth surface, without any aggregation were observed in SEM micrograph of treated powders. Applied treatments are easy, cost-effective and capable to improve functional properties of treated powders that could replace control MPC70 powder in various food applications where protein functionality is of prime importance.

  17. Kinematic reconstitution and tectono-sedimentation associated to salt domes in deep water of Santos basin, Brazil; Reconstituicao cinematica e tectono-sedimentacao associada a domos salinos nas aguas profundas da bacia de Santos, Brasil

    Energy Technology Data Exchange (ETDEWEB)

    Caldas, Manuela Fernandes [PETROBRAS E e P - Exploracao, Rio de Janeiro, RJ (Brazil). Polo Centro], E-mail: manuela.caldas@petrobras.com.br; Zalan, Pedro Victor [PETROBRAS E e P - Exploracao, Rio de Janeiro, RJ (Brazil). Gestao de Projetos Exploratorios], E-mail: zalan@petrobras.com.br

    2009-05-15

    This article presents new methods for structural restoration and kinematic evolution of salt domes during the opening stages and growth of a passive margin in the Santos Basin. The traditional method or restoration of salt bodies uses software and a 3D database. Using 2D seismic data and adapting/improving methods published in the oil industry literature, we developed a method based on the observation of the stratal geometric patterns of sedimentary packages associated to the salt bodies such as tabular forms (straight parallel reflectors, pre or post-tectonic sedimentary sequences), or dish/bowl forms (with very distinct depocenters and thinned margins with fanning of dips, syntectonic/halokinetic sedimentary sequences), and on the determination/mapping of growth axes and thinning axes in halokinetic strata. Horizontal and vertical reconstitutions of the salt flow (inflation, deflation and horizontal flow) were deduced based on isochron (isopach maps in time) and axes maps and flattened seismic sections at the time of deposition of each associated sedimentary sequence. The resulting maps and reconstitutions presented an amazing correlation with the current patterns mapped in the study area. The interpreted tectonic phases of halokinetic evolution, deduced from the produced maps and reconstitutions, showed a good agreement with the regional events that affected the Santos Basin and adjoining basement. Based on those results, we believe that the application of this method provides an important contribution for the oil exploration industry, considering that it presents outstanding results based on 2D seismic data. It allows an understanding of the structural and kinematic evolution of the salt bodies and of the halokinetic sequences associated, with important implications on the modeling of petroleum systems, thus, reducing the risk of exploration prospects. (author)

  18. Functional reconstitution into liposomes of purified human RhCG ammonia channel.

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    Isabelle Mouro-Chanteloup

    Full Text Available BACKGROUND: Rh glycoproteins (RhAG, RhBG, RhCG are members of the Amt/Mep/Rh family which facilitate movement of ammonium across plasma membranes. Changes in ammonium transport activity following expression of Rh glycoproteins have been described in different heterologous systems such as yeasts, oocytes and eukaryotic cell lines. However, in these complex systems, a potential contribution of endogenous proteins to this function cannot be excluded. To demonstrate that Rh glycoproteins by themselves transport NH(3, human RhCG was purified to homogeneity and reconstituted into liposomes, giving new insights into its channel functional properties. METHODOLOGY/PRINCIPAL FINDINGS: An HA-tag introduced in the second extracellular loop of RhCG was used to purify to homogeneity the HA-tagged RhCG glycoprotein from detergent-solubilized recombinant HEK293E cells. Electron microscopy analysis of negatively stained purified RhCG-HA revealed, after image processing, homogeneous particles of 9 nm diameter with a trimeric protein structure. Reconstitution was performed with sphingomyelin, phosphatidylcholine and phosphatidic acid lipids in the presence of the C(12E(8 detergent which was subsequently removed by Biobeads. Control of protein incorporation was carried out by freeze-fracture electron microscopy. Particle density in liposomes was a function of the Lipid/Protein ratio. When compared to empty liposomes, ammonium permeability was increased two and three fold in RhCG-proteoliposomes, depending on the Lipid/Protein ratio (1/300 and 1/150, respectively. This strong NH(3 transport was reversibly inhibited by mercuric and copper salts and exhibited a low Arrhenius activation energy. CONCLUSIONS/SIGNIFICANCE: This study allowed the determination of ammonia permeability per RhCG monomer, showing that the apparent Punit(NH3 (around 1x10(-3 microm(3xs(-1 is close to the permeability measured in HEK293E cells expressing a recombinant human RhCG (1.60x10

  19. [Methodological aspects of the reconstitution and evaluation of the behavioral theories that underlie population policy].

    Science.gov (United States)

    Leeuw, F

    1991-09-01

    This work discusses methodological aspects of the articulation and evaluation of behavioral theories underlying demographic policies. Such theories, called "policy theories" among other terms, may be defined as a group of hypotheses explicitly translated into predictions about behavior that underlie policy measures and that concern the relations between the measure and the objective to be attained. Interest in policy theories has been reflected in the writings of such demographers as D. Bogue, J. Blake, and T. Burch, and of researchers from other social science disciplines. 2 examples of policy theories from the Netherlands are presented to illustrate the discussion, 1 describing family planning communication programs that were intended to reduce the number of unwanted and unplanned pregnancies, and the other describing measures to increase availability of child care services in order to facilitate labor force participation of women and ultimately to increase the birth rate. Both theories are found to be comprised of 2 main parallel theories and several related hypotheses. Because political authorities do not usually make explicit the hypotheses that support political measures, their hypotheses must be articulated and reconstituted through attention to debates, written communications, interviews, and other means. The reconstitution must be done as objectively as possible, which implies the need to follow some methodologic rules. Examples are cited of principles advanced by researchers in management science, market research, and political science. 7 methodological rules or steps are then suggested for articulating policy theories: 1) identify statements relative to the political problem, such as excessive or inadequate fertility rates; 2) use the sources to identify reasons for undertaking concrete policy measures; 3) describe the role of the official in the political process; 4) inventory all declarations concerning the relationship between the objective and the

  20. Molecular genetic analysis of a cattle population to reconstitute the extinct Algarvia breed

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    Rangel-Figueiredo Teresa

    2010-06-01

    Full Text Available Abstract Background Decisions to initiate conservation programmes need to account for extant variability, diversity loss and cultural and economic aspects. Molecular markers were used to investigate if putative Algarvia animals could be identified for use as progenitors in a breeding programme to recover this nearly extinct breed. Methods 46 individuals phenotypically representative of Algarvia cattle were genotyped for 27 microsatellite loci and compared with 11 Portuguese autochthonous and three imported breeds. Genetic distances and factorial correspondence analyses (FCA were performed to investigate the relationship among Algarvia and related breeds. Assignment tests were done to identify representative individuals of the breed. Y chromosome and mtDNA analyses were used to further characterize Algarvia animals. Gene- and allelic-based conservation analyses were used to determine breed contributions to overall genetic diversity. Results Genetic distance and FCA results confirmed the close relationship between Algarvia and southern Portuguese breeds. Assignment tests without breed information classified 17 Algarvia animals in this cluster with a high probability (q > 0.95. With breed information, 30 cows and three bulls were identified (q > 0.95 that could be used to reconstitute the Algarvia breed. Molecular and morphological results were concordant. These animals showed intermediate levels of genetic diversity (MNA = 6.0 ± 1.6, Rt = 5.7 ± 1.4, Ho = 0.63 ± 0.19 and He = 0.69 ± 0.10 relative to other Portuguese breeds. Evidence of inbreeding was also detected (Fis = 0.083, P st = 0.028, P > 0.05. Algarvia cattle provide an intermediate contribution (CB = 6.18, CW = -0.06 and D1 = 0.50 to the overall gene diversity of Portuguese cattle. Algarvia and seven other autochthonous breeds made no contribution to the overall allelic diversity. Conclusions Molecular analyses complemented previous morphological findings to identify 33 animals that

  1. Clinical simulation with dramatization: gains perceived by students and health professionals.

    Science.gov (United States)

    Negri, Elaine Cristina; Mazzo, Alessandra; Martins, José Carlos Amado; Pereira, Gerson Alves; Almeida, Rodrigo Guimarães Dos Santos; Pedersoli, César Eduardo

    2017-08-03

    to identify in the literature the gains health students and professionals perceive when using clinical simulation with dramatization resources. integrative literature review, using the method proposed by the Joanna Briggs Institute (JBI). A search was undertaken in the following databases: Latin American and Caribbean Health Sciences Literature, Web of Science, National Library of Medicine, Cumulative Index to Nursing and Allied Health Literature, The Cochrane Library, Scopus, Scientific Electronic Library Online. 53 studies were analyzed, which complied with the established inclusion criteria. Among the different gains obtained, satisfaction, self-confidence, knowledge, empathy, realism, reduced level of anxiety, comfort, communication, motivation, capacity for reflection and critical thinking and teamwork stand out. the evidence demonstrates the great possibilities to use dramatization in the context of clinical simulation, with gains in the different health areas, as well as interprofessional gains. identificar na literatura quais os ganhos percebidos pelos estudantes e profissionais da área de saúde, utilizando-se da simulação clínica realizada com recursos da dramatização. revisão integrativa da literatura, com a metodologia proposta pelo Instituto Joanna Briggs (JBI), com busca nas bases de dados: Literatura Latino-Americana e do Caribe em Ciências da Saúde, Web of Science, National Library of Medicine, Cumulative Index to Nursing and Allied Health Literature, The Cochrane Library, Scopus, Scientific Electronic Library Online. foram analisados 53 estudos, que atenderam os critérios de inclusão estabelecidos. Entre os diversos ganhos obtidos, destaca-se a satisfação, autoconfiança, conhecimento, empatia, realismo, diminuição do nível de ansiedade, conforto, comunicação, motivação, capacidade de reflexão e de pensamento crítico e trabalho em equipe. as evidências demonstram a ampla possibilidade de uso da dramatização no contexto de

  2. Dramatic loss of Ube3A expression during aging of the mammalian cortex

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    Kate Williams

    2010-05-01

    Full Text Available Neurobiological studies of aging are beginning to link functional changes with a loss of experience-dependent plasticity. In the visual system, age-related functional changes include decreases in visual acuity, orientation selectivity, motion perception, and ocular dominance plasticity. A recent paper has shown that Ube3A, an E3 ubiquitin ligase that is absent in Angelman's Syndrome, is required for experience-dependent plasticity during development of the visual cortex. Knocking out Ube3A during development leads to rigidity of ocular dominance plasticity that is strikingly similar to the reduced plasticity seen in older animals. Furthermore, ubiquitin ligases have been linked with age-related neurodegenerative disorders and longevity. Ubiquitin ligases selectively mark proteins for degradation, and a balance between synaptic proteins and their degradation is important for neural transmission and plasticity. This led us to ask whether normal aging is characterized by a loss of Ube3A in the cortex. We used Western blot analysis in order to quantify Ube3A expression across the life span of humans, macaque monkeys, and cats. We found that Ube3A expression declines across the lifespan in human, monkey, and cat cortex. The losses were substantial (50-80% in all areas studied which includes V1, V3, V4, frontal, and auditory cortex. In addition, when compared with other synaptic proteins there was a selective loss of Ube3A in human cortex. The progressive loss of Ube3A expression during cortical aging is an important new finding. Furthermore, the selective loss of Ube3A in human cortex highlights a specific vulnerability in human brain aging that may signify a dramatic shift in cortical function and plasticity.

  3. Cellulolytic Protist Numbers Rise and Fall Dramatically in Termite Queens and Kings during Colony Foundation

    Science.gov (United States)

    Shimada, Keisuke; Lo, Nathan; Kitade, Osamu; Wakui, Akane

    2013-01-01

    Among the best-known examples of mutualistic symbioses is that between lower termites and the cellulolytic flagellate protists in their hindguts. Although the symbiosis in worker termites has attracted much attention, there have been only a few studies of protists in other castes. We have performed the first examination of protist population dynamics in queens and kings during termite colony foundation. Protist numbers, as well as measurements of hindgut and reproductive tissue sizes, were undertaken at five time points over 400 days in incipient colonies of Reticulitermes speratus, as well as in other castes of mature colonies of this species. We found that protist numbers increased dramatically in both queens and kings during the first 50 days of colony foundation but began to decrease by day 100, eventually disappearing by day 400. Hindgut width followed a pattern similar to that of protist numbers, while ovary and testis widths increased significantly only at day 400. Kings were found to contain higher numbers of protists than queens in incipient colonies, which may be linked to higher levels of nutrient transfer from kings to queens than vice versa, as is known in some other termite species. Protists were found to be abundant in soldiers from mature colonies but absent in neotenics. This probably reflects feeding of soldiers by workers via proctodeal trophallaxis and of reproductives via stomodeal trophallaxis. The results reveal the dynamic nature of protist numbers during colony foundation and highlight the trade-offs that exist between reproduction and parental care during this critical phase of the termite life cycle. PMID:23376945

  4. Chemical modification of lysine residues in lysozyme may dramatically influence its amyloid fibrillation.

    Science.gov (United States)

    Morshedi, Dina; Ebrahim-Habibi, Azadeh; Moosavi-Movahedi, Ali Akbar; Nemat-Gorgani, Mohsen

    2010-04-01

    Studies on the aggregation of mutant proteins have provided new insights into the genetics of amyloid diseases and the role of the net charge of the protein on the rate, extent, and type of aggregate formation. In the present work, hen egg white lysozyme (HEWL) was employed as the model protein. Acetylation and (separately) citraconylation were employed to neutralize the charge on lysine residues. Acetylation of the lysine residues promoted amyloid formation, resulting in more pronounced fibrils and a dramatic decline in the nucleation time. In contrast, citraconylation produced the opposite effect. In both cases, native secondary and tertiary structures appeared to be retained. Studies on the effect of pH on aggregation suggested greater possibilities for amorphous aggregate formation rather than fibrillation at pH values closer to neutrality, in which the protein is known to take up a conformation more similar to its native form. This is in accord with reports in the literature suggesting that formation of amorphous aggregates is more favored under relatively more native conditions. pH 5 provided a critical environment in which a mixture of amorphous and fibrillar structures were observed. Use of Tango and Aggrescan software which describe aggregation tendencies of different parts of a protein structure suggested critical importance of some of the lysine residues in the aggregation process. Results are discussed in terms of the importance of the net charge in control of protein-protein interactions leading to aggregate formation and possible specific roles of lysine residues 96 and 97. Copyright 2009 Elsevier B.V. All rights reserved.

  5. Early NK Cell Reconstitution Predicts Overall Survival in T-Cell Replete Allogeneic Hematopoietic Stem Cell Transplantation

    DEFF Research Database (Denmark)

    Minculescu, Lia; Marquart, Hanne Vibeke; Friis, Lone Smidstrups

    2016-01-01

    Early immune reconstitution plays a critical role in clinical outcome after allogeneic hematopoietic stem cell transplantation (HSCT). Natural killer (NK) cells are the first lymphocytes to recover after transplantation and are considered powerful effector cells in HSCT. We aimed to evaluate...... the clinical impact of early NK cell recovery in T-cell replete transplant recipients. Immune reconstitution was studied in 298 adult patients undergoing HSCT for acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL) and myelodysplastic syndrome (MDS) from 2005 to 2013. In multivariate analysis NK...... cell numbers day 30 (NK30) >150cells/µL were independently associated with superior overall survival (hazard ratio 0.79, 95% confidence interval 0.66-0.95, p=0.01). Cumulative incidence analyses showed that patients with NK30 >150cells/µL had significantly less transplant related mortality (TRM), p=0...

  6. Insights into pathogenic events of HIV-associated Kaposi sarcoma and immune reconstitution syndrome related Kaposi sarcoma

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    Lemmer Johan

    2008-01-01

    Full Text Available Abstract A decrease in the incidence of human immune deficiency virus-associated Kaposi sarcoma (HIV-KS and regression of some established HIV-KS lesions is evident after the introduction of highly active anti-retroviral treatment (HAART, and is attributed to generalized immune restoration, to the reconstitution of human herpesvirus (HHV-8 specific cellular immune responses, and to the decrease in HIV Tat protein and HHV-8 loads following HAART. However, a small subset of HIV-seropositive subjects with a low CD4+ T cell count at the time of introduction of HAART, may develop HIV-KS as immune reconstitution inflammatory syndrome (IRIS within 8 weeks thereafter.

  7. Comparative Analysis of Stress Induced Gene Expression in Caenorhabditis elegans following Exposure to Environmental and Lab Reconstituted Complex Metal Mixture.

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    Ranjeet Kumar

    Full Text Available Metals are essential for many physiological processes and are ubiquitously present in the environment. However, high metal concentrations can be harmful to organisms and lead to physiological stress and diseases. The accumulation of transition metals in the environment due to either natural processes or anthropogenic activities such as mining results in the contamination of water and soil environments. The present study used Caenorhabditis elegans to evaluate gene expression as an indicator of physiological response, following exposure to water collected from three different locations downstream of a Swedish mining site and a lab reconstituted metal mixture. Our results indicated that the reconstituted metal mixture exerted a direct stress response in C. elegans whereas the environmental waters elicited either a diminished or abrogated response. This suggests that it is not sufficient to use the biological effects observed from laboratory mixtures to extrapolate the effects observed in complex aquatic environments and apply this to risk assessment and intervention.

  8. Reconstitution radicicol containing apolipoprotein B lipoparticle and tracing its cell uptake process by super resolution fluorescent microscopy.

    Science.gov (United States)

    Lin, Chung Ching; Lin, Po-Yen; Chang, Chia-Ching

    Apolipoprotein B (apoB) is the only protein of LDL. LDL delivers cholesterol, triacylglycerides and lipids to the target cells. Reconstitute apoB lipoparticle (rABL) will be an idea drug delivery vehicle for hydrophobic and amphiphilic materials delivery. It is challenged to renature ApoB into its functional state from denatured state. By using modified bile salt and radicicol (Rad) added over-critical refolding process, apoB can be restored into its native like state. The intrinsic fluorescence of apoB increased during the refolding process. Moreover, radicicol (Rad) molecules have been encapsulated into reconstitute rABL (Rad@rABL). To investigate the cell uptake mechanism of Rad@rABL, a super resolution ground state depletion (GSD) microscopy is used in this research. Fluorescence labeled Rad@rABL can be traced within the tumor cell. Key words: LDL, radicicol, protein refolding, super resolution microscopy.

  9. Regulatory T Cells in HIV-Infected Immunological Nonresponders Are Increased in Blood but Depleted in Lymphoid Tissue and Predict Immunological Reconstitution

    DEFF Research Database (Denmark)

    Gaardbo, Julie C; Hartling, Hans J; Ronit, Andreas

    2014-01-01

    BACKGROUND: HIV-infected immunological nonresponders fail to immune reconstitute despite optimal treatment. We hypothesized that regulatory T cells (Tregs) are involved in immunological reconstitution. Tregs and Treg subpopulations were measured in blood and Foxp3 cells in lymphoid tissue......, and the impact of Tregs on immunological reconstitution was determined. METHODS: HIV-infected individuals on combination antiretroviral therapy for a minimum of 2 years were included. The study population included 14 immunological nonresponders (INR; CD4 T-cell count .... In contrast, responders resembled healthy controls. Finally, in INR, high level of Tregs in blood and Foxp3 cells in lymphoid tissue were associated with higher level of immunological reconstitution after 1 year of follow-up. CONCLUSIONS: In conclusion, altered distribution of Tregs was found in INR...

  10. Interaction between sodium dodecyl sulfate and membrane reconstituted aquaporins: A comparative study of spinach SoPIP2;1 and E. coli AqpZ

    DEFF Research Database (Denmark)

    Hansen, Jesper Schmidt; Vararattanavech, Ardcharaporn; Plasencia, Inés

    2011-01-01

    This study describes the interaction between sodium dodecyl sulfate (SDS) and membrane proteins reconstituted into large unilamellar lipid vesicles and detergent micelles studied by circular dichroism (CD) and polarity sensitive probe labeling. Specifically, we carried out a comparative study of ...

  11. Dynamical System Modeling of Immune Reconstitution after Allogeneic Stem Cell Transplantation Identifies Patients at Risk for Adverse Outcomes.

    Science.gov (United States)

    Toor, Amir A; Sabo, Roy T; Roberts, Catherine H; Moore, Bonny L; Salman, Salman R; Scalora, Allison F; Aziz, May T; Shubar Ali, Ali S; Hall, Charles E; Meier, Jeremy; Thorn, Radhika M; Wang, Elaine; Song, Shiyu; Miller, Kristin; Rizzo, Kathryn; Clark, William B; McCarty, John M; Chung, Harold M; Manjili, Masoud H; Neale, Michael C

    2015-07-01

    Systems that evolve over time and follow mathematical laws as they evolve are called dynamical systems. Lymphocyte recovery and clinical outcomes in 41 allograft recipients conditioned using antithymocyte globulin (ATG) and 4.5-Gy total body irradiation were studied to determine if immune reconstitution could be described as a dynamical system. Survival, relapse, and graft-versus-host disease (GVHD) were not significantly different in 2 cohorts of patients receiving different doses of ATG. However, donor-derived CD3(+) cell reconstitution was superior in the lower ATG dose cohort, and there were fewer instances of donor lymphocyte infusion (DLI). Lymphoid recovery was plotted in each individual over time and demonstrated 1 of 3 sigmoid growth patterns: Pattern A (n = 15) had rapid growth with high lymphocyte counts, pattern B (n = 14) had slower growth with intermediate recovery, and pattern C (n = 10) had poor lymphocyte reconstitution. There was a significant association between lymphocyte recovery patterns and both the rate of change of donor-derived CD3(+) at day 30 after stem cell transplantation (SCT) and clinical outcomes. GVHD was observed more frequently with pattern A, relapse and DLI more so with pattern C, with a consequent survival advantage in patients with patterns A and B. We conclude that evaluating immune reconstitution after SCT as a dynamical system may differentiate patients at risk of adverse outcomes and allow early intervention to modulate that risk. Copyright © 2015 American Society for Blood and Marrow Transplantation. Published by Elsevier Inc. All rights reserved.

  12. Multiphasic and multifocal cryptococcal immune reconstitution inflammatory syndrome in an HIV-infected patient: interplay of infection and immunity.

    Science.gov (United States)

    Katchanov, Juri; Zimmermann, Ulrike; Branding, Gordian; Tintelnot, Kathrin; Müller, Markus; Arastéh, Keikawus; Stocker, Hartmut

    2014-01-01

    We report a case of cryptococcal immune reconstitution inflammatory syndrome affecting the lungs, and 10 months later the cervical lymph nodes, in the absence of cryptococcal meningitis, in advanced HIV infection. Our report demonstrates the organ-specificity of the timing of the inflammatory response and illustrates the organ-specific interplay of immunity and infection in cryptococcal disease. Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.

  13. Different profiles of immune reconstitution in children and adults with HIV-infection after highly active antiretroviral therapy

    Directory of Open Access Journals (Sweden)

    Leal Manuel

    2006-07-01

    Full Text Available Abstract Background Recent advances in characterizing the immune recovery of HIV-1-infected people have highlighted the importance of the thymus for peripheral T-cell diversity and function. The aim of this study was to investigate differences in immune reconstitution profiles after highly active antiretroviral therapy (HAART between HIV-children and adults. Methods HIV patients were grouped according to their previous clinical and immunological status: 9 HIV-Reconstituting-adults (HIV-Rec-adults and 10 HIV-Reconstituting-children (HIV-Rec-children on HAART with viral load (VL ≤400 copies/ml and CD4+ ≥500 cells/μL at least during 6 months before the study and CD4+ ≤300 cells/μL anytime before. Fifteen healthy-adults and 20 healthy-children (control subjects were used to calculate Z-score values to unify value scales between children and adults to make them comparable. Results HIV-Rec-children had higher T-cell receptor excision circles (TREC and lower interleukin (IL-7 levels than HIV-Rec-adults (p + (CD4+CD45RA hi+CD27+, naïve CD8+ (CD8+CD45RA hi+CD27+, and memory CD8+ (CD8+CD45RO+ cells/μl than HIV-Rec-adults, but similar memory CD4+ (CD4+CD45RO+ counts. HIV-Rec-children had lower naïve CD8+ Z-score values than HIV-Rec-adults (p = 0.05. Conclusion Our data suggest that HIV-Rec-children had better thymic function than HIV-Rec-adults and this fact affects the peripheral T-cell subsets. Thus, T-cell recovery after HAART in HIV-Rec-adults could be the consequence of antigen-independent peripheral T-cell expansion while in HIV-Rec-children thymic output could play a predominant role in immune reconstitution.

  14. Stem and stromal cell reconstitution of lethally irradiated mice following transplantation of hematopoietic tissue from donors of various ages

    International Nuclear Information System (INIS)

    Schmidt, C.M.; Doran, G.A.; Crouse, D.A.; Sharp, J.G.

    1987-01-01

    If the limited life span of hematopoietic tissues in vitro is due to a finite proliferative capacity of individual stem cells, one might expect tissues of young donors to possess a greater proliferative capacity and to contain a larger population of primitive stem cells than those of older donors. To test this hypothesis, we used 12- and 8-day spleen colony formation (CFU-s) to assay more and less primitive stem cell subpopulations of three murine hematopoietic tissues: fetal liver (FL) and weanling (WBM) and adult (ABM) bone marrow. Subsequently, the same assays and a stromal cell assay were performed on the bone marrow from groups of lethally irradiated mice reconstituted with these tissues. Comparison of the CFU-s content of the donor tissues revealed that FL contained a significantly greater proportion of primitive stem cells as evidenced by a (Day 12):(Day 8) CFU-s ratio of 3.0 +/- 1.0 as compared to 0.9 +/- 0.1 for WBM and ABM. In addition, at 21 weeks post-transplantation the CFU-s/femur values of the FL reconstituted group were significantly greater than those of the ABM and WBM reconstituted groups. These results suggest that fetal hematopoietic tissue contains a greater proportion of primitive stem cells and has a greater proliferative potential than hematopoietic tissue from older donors. No differences were seen in stromal cell reconstitution of the three experimental groups. In all cases, assayable fibroblast colony forming cells (CFU-f) remained at 20-40% of control values, even at 21 weeks postreconstitution

  15. Development of a reconstitution system of Charpy probes for the surveillance of vessels in nucleo electric plants

    International Nuclear Information System (INIS)

    Romero C, J.; Hernandez, R.; Fernandez, F.; Gonzalez M, A.

    2007-01-01

    This work describes the development of a welding system, for the rebuilding of halves of Charpy test tubes, the rebuilding consists on welding two implants in those ends of these halves of test tubes, in these welding the main requirement is not to alter the mechanical properties in a minimum volume of 1 cm 3 , the rebuilding is medullary in the surveillance programs of the reactor vessel. In these programs, the mechanical state of the vessel is evaluated, for it there are surveillance capsules with a Charpy witness test tubes series, subjected to a neutron flow similar or bigger to that of the vessel. The objective is to evaluate in advance on the vessel fragilization grade its life design. However the number of capsules with the witness test tubes it is only for the plant design life and at the moment the nucleo electric, negotiates an extension of life of these, until for 20 more years, of there the importance of this material witness's that stores the information of the damage accumulated by the neutron flow. This material requires to be taken advantage it after being rehearsed and the normative one settles down as obligatory to qualify the rebuilding process with all the requirements settled down in the ASTM Designation: E 1253-99 'Standard Guide for Reconstitution of irradiated Charpy-Sized Specimens', to obtain other reconstituted Charpy test tubes that are again introduced in the reactor. When being reconstituted the halves of the original test tubes it is obtained double reconstituted Charpy test tubes. Half of the test tubes they are used in the surveillance program of the vessel, with the surpluses test tubes, it can determine the fracture toughness, property of the material used in the extension methodology of life of vessel. (Author)

  16. Purification and functional reconstitution of a seven-subunit mrp-type na+/h+ antiporter.

    Science.gov (United States)

    Morino, Masato; Suzuki, Toshiharu; Ito, Masahiro; Krulwich, Terry Ann

    2014-01-01

    Mrp antiporters and their homologues in the cation/proton antiporter 3 family of the Membrane Transporter Database are widely distributed in bacteria. They have major roles in supporting cation and cytoplasmic pH homeostasis in many environmental, extremophilic, and pathogenic bacteria. These antiporters require six or seven hydrophobic proteins that form hetero-oligomeric complexes, while most other cation/proton antiporters require only one membrane protein for their activity. The resemblance of three Mrp subunits to membrane-embedded subunits of the NADH:quinone oxidoreductase of respiratory chains and to subunits of several hydrogenases has raised interest in the evolutionary path and commonalities of their proton-translocating domains. In order to move toward a greater mechanistic understanding of these unusual antiporters and to rigorously demonstrate that they function as secondary antiporters, powered by an imposed proton motive force, we established a method for purification and functional reconstitution of the seven-subunit Mrp antiporter from alkaliphilic Bacillus pseudofirmus OF4. Na(+)/H(+) antiporter activity was demonstrated by a fluorescence-based assay with proteoliposomes in which the Mrp complex was coreconstituted with a bacterial FoF1-ATPase. Proton pumping by the ATPase upon addition of ATP generated a proton motive force across the membranes that powered antiporter activity upon subsequent addition of Na(+).

  17. An exploratory study on the utilisation of resilience by middle adolescents in reconstituted families following divorce

    Directory of Open Access Journals (Sweden)

    Suzette Ebersohn

    2013-01-01

    Full Text Available Every year thousands of core families disintegrate through divorce, and in the ensuing restructuring of the family system the child has to cope with various development challenges, such as divided membership of two micro family systems and complexities that result at the mesosystemic level. Achieving positive development outcomes in the presence of challenging living circumstances entails complex interactive processes. The aim of the study was to understand the concomitant, reciprocal and/or responsive dynamics of middle adolescents' use of their inherent resilience potential in their movement back and forth between their two reconstituted family systems after the parents' divorce. The study was grounded in the qualitative interpretivist paradigm, and used a multiple case study as research design and a narrative format for description. A purposive sample of four white Afrikaans-speaking middle adolescents participated in the research. Findings revealed that middle adolescents of divorced parents utilise their resilience potential in a systemic manner, which requires a solid base provided by the meso system. Hence the utilisation of resilience relies at the very minimum on a functional relationship of cooperation between the biological parents.

  18. ESCRT-III mediated cell division in Sulfolobus acidocaldarius –A reconstitution perspective

    Directory of Open Access Journals (Sweden)

    Tobias eHärtel

    2014-06-01

    Full Text Available In the framework of Synthetic Biology, it has become an intriguing question what would be the minimal representation of cell division machinery. Thus, it seems appropriate to compare how cell division is realized in different microorganisms. In particular, the cell division system of Crenarchaeota lacks certain proteins found in most bacteria and Euryarchaeota, such as FtsZ, MreB or the Min system. The Sulfolobaceae family encodes functional homologs of the eukaryotic proteins Vps4 and ESCRT-III. ESCRT-III is essential for several eukaryotic pathways, e.g. budding of intralumenal vesicles (ILVs, or cytokinesis, whereas Vps4 dissociates the ESCRT-III complex from the membrane. CdvA (Cell Division A is required for the recruitment of crenarchaeal ESCRT-III proteins to the membrane at mid-cell. The proteins polymerize and form a smaller structure during constriction. Thus, ESCRT-III mediated cell division in S. acidocaldarius shows functional analogies to the Z ring observed in prokaryotes like E. coli, which has recently begun to be reconstituted in vitro. In this short perspective, we discuss the possibility of building such an in vitro cell division system on basis of archaeal ESCRT-III.

  19. Studying Mechanosensitivity of Two-Pore Domain K+ Channels in Cellular and Reconstituted Proteoliposome Membranes.

    Science.gov (United States)

    Del Mármol, Josefina; Rietmeijer, Robert A; Brohawn, Stephen G

    2018-01-01

    Mechanical force sensation is fundamental to a wide breadth of biology from the classic senses of touch, pain, hearing, and balance to less conspicuous sensations of proprioception, blood pressure, and osmolarity and basic aspects of cell growth, differentiation, and development. These diverse and essential systems use force-gated (or mechanosensitive) ion channels that convert mechanical stimuli into cellular electrical signals. TRAAK, TREK1, and TREK2 are K + -selective ion channels of the two-pore domain K + (K2P) family that are mechanosensitive: they are gated open by increasing membrane tension. TRAAK and TREK channels are thought to play roles in somatosensory and other mechanosensory processes in neuronal and non-neuronal tissues. Here, we present protocols for three assays to study mechanical activation of these channels in cell membranes: (1) cell swelling, (2) cell poking, and (3) patched membrane stretching. Patched membrane stretching is also applicable to the study of mechanosensitive K2P channel activity in a cell-free system and a procedure for proteoliposome reconstitution and patching is also presented. These approaches are also readily applicable to the study of other mechanosensitive ion channels.

  20. Stabilization and immobilization of aquaporin reconstituted lipid vesicles for water purification.

    Science.gov (United States)

    Sun, Guofei; Chung, Tai-Shung; Jeyaseelan, Kandiah; Armugam, Arunmozhiarasi

    2013-02-01

    Aquaporins are water channel proteins in biological membranes that have extraordinary water permeability and selectivity. In this work, we have demonstrated that one of their family members, AquaporinZ (AqpZ), can be possibly applied in a pressure-driven water purification process. A nanofiltration membrane was designed and fabricated by immobilization of AqpZ-reconstituted liposomes on a polydopamine (PDA) coated microporous membrane. Amine-functionalized proteoliposomes were first deposited via gentle vacuum suction and subsequently conjugated on the PDA layer via an amine-catechol adduct formation. Due to the existence of a polymer network within the lipid bilayers, the membrane could sustain hydraulic pressure of 5 bar as well as the strong surface agitation in nanofiltration tests, indicating a relatively stable membrane structure. In comparison with membrane without AqpZ incorporation, the membrane with AqpZ-to-lipid weight ratio of 1:100 increased the water flux by 65% with enhanced NaCl and MgCl(2) rejections of 66.2% and 88.1%, respectively. With AqpZ incorporation, the vesicle immobilized membrane exhibits a promising strategy for high productivity water purification. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Constitutive dimerization of the G-protein coupled receptor, neurotensin receptor 1, reconstituted into phospholipid bilayers.

    Science.gov (United States)

    Harding, Peter J; Attrill, Helen; Boehringer, Jonas; Ross, Simon; Wadhams, George H; Smith, Eleanor; Armitage, Judith P; Watts, Anthony

    2009-02-01

    Neurotensin receptor 1 (NTS1), a Family A G-protein coupled receptor (GPCR), was expressed in Escherichia coli as a fusion with the fluorescent proteins eCFP or eYFP. A fluorophore-tagged receptor was used to study the multimerization of NTS1 in detergent solution and in brain polar lipid bilayers, using fluorescence resonance energy transfer (FRET). A detergent-solubilized receptor was unable to form FRET-competent complexes at concentrations of up to 200 nM, suggesting that the receptor is monomeric in this environment. When reconstituted into a model membrane system at low receptor density, the observed FRET was independent of agonist binding, suggesting constitutive multimer formation. In competition studies, decreased FRET in the presence of untagged NTS1 excludes the possibility of fluorescent protein-induced interactions. A simulation of the experimental data indicates that NTS1 exists predominantly as a homodimer, rather than as higher-order multimers. These observations suggest that, in common with several other Family A GPCRs, NTS1 forms a constitutive dimer in lipid bilayers, stabilized through receptor-receptor interactions in the absence of other cellular signaling components. Therefore, this work demonstrates that well-characterized model membrane systems are useful tools for the study of GPCR multimerization, allowing fine control over system composition and complexity, provided that rigorous control experiments are performed.

  2. Evaluation of apple pomace based reconstituted feed in rats after solid state fermentation and ethanol recovery

    Directory of Open Access Journals (Sweden)

    A. Devrajan

    2004-03-01

    Full Text Available To utilize apple pomace in an economical and effective way, a feed was developed by solid state fermentation (SSF using sequential interactive co-culture of Candida utilis and Kloeckera. Removal of ethanol and drying of the left - over residue. Feeding trial was conducted in white albino rats before and after reconstitution of apple pomace feed in the choice and no choice study. Feeding of apple pomace feed in the rats before reconstitution indicated that neither in fermented nor in unfermented form it was acceptable. In the no choice study both in 100% fermented and unfermented apple pomace group feed intake decreased continuously resulting in death of rats apparently due to decreased digestibility owing to high fibre content. Further, fermented or unfermented apple pomace based feed had lower digestibility and efficiency of conversion than the standard rat feed. Compared to the standard feed group, growth rates in all the feed groups were negative. The results of choice study of different feeds corroborated with the no-choice study mentioned earlier. Incorporation of fermented apple pomace into standard rat feed in the ratio of 1:1 gave better acceptability and digestibility. Reconstituted feed with 10% jaggery, 2% groundnut oil, 0.01% mixed flavour and 1% salt was the most acceptable. The post-mortem examination of the rats that died during the feeding trial revealed generalized oedema, probably due to some hepatotoxin in the feed. The blood glucose level in the apple pomace feed group except that was reconstituted with jaggery, showed a general hypoglycemia, though falling in the range. The increased ALT and AST levels in the serum also suggest damage to the liver. The mortality rate, post-mortem examination of the rats that died during the feeding trial and blood biochemical analysis of rat serum suggest a more elaborative study for extended period of time.Para utilizar bagaço de maçã de uma maneira econômica e eficaz, foi desenvolvido

  3. Reconstitution of high-affinity opioid agonist binding in brain membranes

    Energy Technology Data Exchange (ETDEWEB)

    Remmers, A.E.; Medzihradsky, F. (Univ. of Michigan Medical School, Ann Arbor (United States))

    1991-03-15

    In synaptosomal membranes from rat brain cortex, the {mu} selective agonist ({sup 3}H)dihydromorphine in the absence of sodium, and the nonselective antagonist ({sup 3}H)naltrexone in the presence of sodium, bound to two populations of opioid receptor sites with K{sub d} values of 0.69 and 8.7 nM for dihydromorphine, and 0.34 and 5.5 nM for naltrexone. The addition of 5 {mu}M guanosine 5{prime}-({gamma}-thio)triphosphate (GTP({gamma}S)) strongly reduced high-affinity agonist but not antagonist binding. Exposure of the membranes to high pH reduced the number of GTP({gamma}-{sup 35}S) binding sites by 90% and low K{sub m}, opioid-sensitive GTPase activity by 95%. In these membranes, high-affinity agonist binding was abolished and modulation of residual binding by GTP({gamma}S) was diminished. Alkali treatment of the glioma cell membranes prior to fusion inhibited most of the low K{sub m} GTPase activity and prevented the reconstitution of agonist binding. The results show that high-affinity opioid agonist binding reflects the ligand-occupied receptor - guanine nucleotide binding protein complex.

  4. Intensive fibrosarcoma-binding capability of the reconstituted analog and its antitumor activity.

    Science.gov (United States)

    Xu, Jian; Du, Yue; Liu, Wen-Juan; Li, Liang; Li, Yi; Wang, Xiao-Fei; Yi, Hong-Fei; Shan, Chuan-Kun; Xia, Gui-Min; Liu, Xiu-Jun; Zhen, Yong-Su

    2018-11-01

    Fibrosarcomas are highly aggressive malignant tumors. It is urgently needed to explore targeted drugs and modalities for more effective therapy. Matrix metalloproteinases (MMPs) play important roles in tumor progression and metastasis, while several MMPs are highly expressed in fibrosarcomas. In addition, tissue inhibitor of metalloproteinase 2 (TIMP2) displays specific interaction with MMPs. Therefore, TIMP2 may play an active role in the development of fibrosarcoma-targeting agents. In the current study, a TIMP2-based recombinant protein LT and its enediyne-integrated analog LTE were prepared; furthermore, the fibrosarcoma-binding intensity and antitumor activity were investigated. As shown, intense and selective binding capability of the protein LT to human fibrosarcoma specimens was confirmed by tissue microarray. Moreover, LTE, the enediyne-integrated analog of LT, exerted highly potent cytotoxicity to fibrosarcoma HT1080 cells, induced apoptosis, and caused G2/M arrest. LTE at 0.1 nM markedly suppressed the migration and invasion of HT1080 cells. LTE at tolerated dose of 0.6 mg/kg inhibited the tumor growth of fibrosarcoma xenograft in athymic mice. The study provides evidence that the TIMP2-based reconstituted analog LTE may be useful as a targeted drug for fibrosarcome therapy.

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

    Science.gov (United States)

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

    2014-11-01

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

  6. Characterization of Cu(II)-reconstituted ACC Oxidase using experimental and theoretical approaches.

    Science.gov (United States)

    El Bakkali-Tahéri, Nadia; Tachon, Sybille; Orio, Maylis; Bertaina, Sylvain; Martinho, Marlène; Robert, Viviane; Réglier, Marius; Tron, Thierry; Dorlet, Pierre; Simaan, A Jalila

    2017-06-01

    1-Aminocyclopropane-1-carboxylic acid oxidase (ACCO) is a non heme iron(II) containing enzyme that catalyzes the final step of the ethylene biosynthesis in plants. The iron(II) ion is bound in a facial triad composed of two histidines and one aspartate (H177, D179 and H234). Several active site variants were generated to provide alternate binding motifs and the enzymes were reconstituted with copper(II). Continuous wave (cw) and pulsed Electron Paramagnetic Resonance (EPR) spectroscopies as well as Density Functional Theory (DFT) calculations were performed and models for the copper(II) binding sites were deduced. In all investigated enzymes, the copper ion is equatorially coordinated by the two histidine residues (H177 and H234) and probably two water molecules. The copper-containing enzymes are inactive, even when hydrogen peroxide is used in peroxide shunt approach. EPR experiments and DFT calculations were undertaken to investigate substrate's (ACC) binding on the copper ion and the results were used to rationalize the lack of copper-mediated activity. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Purification and reconstitution of the calcium antagonist receptor of the voltage-sensitive calcium channel

    International Nuclear Information System (INIS)

    Curtis, B.M.

    1986-01-01

    Treatment with digitonin solubilized the calcium antagonist receptor as a stable complex with [ 3 H]nitrendipine from rat brain membranes. The solubilized complex retains allosteric coupling to binding sites for diltiazem, verapamil, and inorganic calcium antagonist sites. The calcium antagonist receptor from cardiac sarcolemma and the transverse-tubule membrane of skeletal muscle is also efficiently solubilized with digitonin and the receptor in all three tissues is a large glycoprotein with a sedimentation coefficient of 20 S. The T-tubule calcium antagonist receptor complex was extensively purified by a combination of chromatography on WGA-Sepharose, ion exchange chromatography, and sedimentation on sucrose gradients to yield preparations estimated to be 41% homogeneous by specific activity and 63% homogeneous by SDS gel electrophoresis. Analysis of SDS gels detect three polypeptides termed α(Mr 135,000), β(Mr 50,000), and γ(Mr 32,000) as noncovalently associated subunits of the calcium antagonist receptor. The α and γ subunits are glycosylated polypeptides, and the molecular weight of the core polypeptides are 108,000 and 24,000 respectively. The calcium antagonist receptor was reconstituted into a phospholipid bilayer by adding CHAPS and exogeneous lipid to the purified receptor followed by rapid detergent removal. This procedure resulted in the incorporation of 45% of the calcium antagonist receptor into closed phospholipid vesicles. Data suggests that the α, β, and γ subunits of the T-tubule calcium antagonist receptor are sufficient to form a functional calcium channel

  8. Centromeric DNA replication reconstitution reveals DNA loops and ATR checkpoint suppression.

    Science.gov (United States)

    Aze, Antoine; Sannino, Vincenzo; Soffientini, Paolo; Bachi, Angela; Costanzo, Vincenzo

    2016-06-01

    Half of the human genome is made up of repetitive DNA. However, mechanisms underlying replication of chromosome regions containing repetitive DNA are poorly understood. We reconstituted replication of defined human chromosome segments using bacterial artificial chromosomes in Xenopus laevis egg extract. Using this approach we characterized the chromatin assembly and replication dynamics of centromeric alpha-satellite DNA. Proteomic analysis of centromeric chromatin revealed replication-dependent enrichment of a network of DNA repair factors including the MSH2-6 complex, which was required for efficient centromeric DNA replication. However, contrary to expectations, the ATR-dependent checkpoint monitoring DNA replication fork arrest could not be activated on highly repetitive DNA due to the inability of the single-stranded DNA binding protein RPA to accumulate on chromatin. Electron microscopy of centromeric DNA and supercoil mapping revealed the presence of topoisomerase I-dependent DNA loops embedded in a protein matrix enriched for SMC2-4 proteins. This arrangement suppressed ATR signalling by preventing RPA hyper-loading, facilitating replication of centromeric DNA. These findings have important implications for our understanding of repetitive DNA metabolism and centromere organization under normal and stressful conditions.

  9. Inactivation of Enterobacter aerogenes in reconstituted skim milk by high- and low-frequency ultrasound.

    Science.gov (United States)

    Gao, Shengpu; Hemar, Yacine; Lewis, Gillian D; Ashokkumar, Muthupandian

    2014-11-01

    The inactivation of Enterobacter aerogenes in skim milk using low-frequency (20kHz) and high-frequency (850kHz) ultrasonication was investigated. It was found that low-frequency acoustic cavitation resulted in lethal damage to E. aerogenes. The bacteria were more sensitive to ultrasound in water than in reconstituted skim milk having different protein concentrations. However, high-frequency ultrasound was not able to inactivate E. aerogenes in milk even when powers as high as 50W for 60min were used. This study also showed that high-frequency ultrasonication had no influence on the viscosity and particle size of skim milk, whereas low-frequency ultrasonication resulted in the decrease in viscosity and particle size of milk. The decrease in particle size is believed to be due to the breakup of the fat globules, and possibly to the cleavage of the κ-casein present at the surface of the casein micelles. Whey proteins were also found to be slightly affected by low-frequency ultrasound, with the amounts of α-lactalbumin and β-lactoglobulin slightly decreasing. Copyright © 2013. Published by Elsevier B.V.

  10. Engineered Murine HSCs Reconstitute Multi-lineage Hematopoiesis and Adaptive Immunity

    Directory of Open Access Journals (Sweden)

    Yi-Fen Lu

    2016-12-01

    Full Text Available Hematopoietic stem cell (HSC transplantation is curative for malignant and genetic blood disorders, but is limited by donor availability and immune-mismatch. Deriving HSCs from patient-matched embryonic/induced-pluripotent stem cells (ESCs/iPSCs could address these limitations. Prior efforts in murine models exploited ectopic HoxB4 expression to drive self-renewal and enable multi-lineage reconstitution, yet fell short in delivering robust lymphoid engraftment. Here, by titrating exposure of HoxB4-ESC-HSC to Notch ligands, we report derivation of engineered HSCs that self-renew, repopulate multi-lineage hematopoiesis in primary and secondary engrafted mice, and endow adaptive immunity in immune-deficient recipients. Single-cell analysis shows that following engraftment in the bone marrow niche, these engineered HSCs further specify to a hybrid cell type, in which distinct gene regulatory networks of hematopoietic stem/progenitors and differentiated hematopoietic lineages are co-expressed. Our work demonstrates engineering of fully functional HSCs via modulation of genetic programs that govern self-renewal and lineage priming.

  11. Immune reconstitution with two different rabbit polyclonal anti-thymocytes globulins.

    Science.gov (United States)

    Bamoulid, Jamal; Crepin, Thomas; Gaiffe, Emilie; Laheurte, Caroline; Moulin, Bruno; Frimat, Luc; Rieu, Philippe; Mousson, Christiane; Durrbach, Antoine; Heng, Anne-Elisabeth; Rebibou, Jean-Michel; Saas, Philippe; Courivaud, Cécile; Ducloux, Didier

    2017-12-01

    Broad T cell depletion by polyclonal anti-thymocyte globulins (ATG) has been used for many years as a part of immunosuppressive treatment in transplantation. Currently, two different ATG are used in clinical practice, Thymoglobulin and Grafalon. Due to differences in the immunization source, these products contain different specificities and quantity of antibodies. These differences may have clinical consequences. We conducted a nested study in a large prospective multicentric cohort of kidney transplant to determine whether Grafalon-treated and Thymoglobulin-treated patients experience different lymphocyte reconstitution and clinical outcomes. 182 patients matched for age, gender, CMV status, CMV prophylaxis, number of previous transplantation, and maintenance immunosuppressive treatment were included (Thymoglobulin, [n=91]; Grafalon®, [n=91]). One-year post-transplant, recent thymic emigrants were significantly decreased (12±10% vs 21±12%; p<0.001) in Grafalon-treated patients. By contrast, T cell activation (CD38+DR+Ki67+) and senescence (CD8+CD57+CD28-) was increased in Thymoglobulin-treated patients. Compared to Grafalon, Thymoglobulin was not associated with a significantly different rate of acute rejection. CMV disease (p=0.013) was more frequent in Thymoglobulin-treated patients. Grafalon and Thymoglobulin seem to be equivalent to prevent acute rejection. CMV disease is more frequent in Thymoglobulin-treated patients. One year post-transplant immune profile profoundly differs according to the type of ATG. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. A non-equilibrium thermodynamics model of reconstituted Ca(2+)-ATPase.

    Science.gov (United States)

    Waldeck, A R; van Dam, K; Berden, J; Kuchel, P W

    1998-01-01

    A non-equilibrium thermodynamics (NET) model describing the action of completely coupled or 'slipping' reconstituted Ca(2+)-ATPase is presented. Variation of the coupling stoichiometries with the magnitude of the electrochemical gradients, as the ATPase hydrolyzes ATP, is an indication of molecular slip. However, the Ca2+ and H+ membrane-leak conductances may also be a function of their respective gradients. Such non-ohmic leak typically yields 'flow-force' relationships that are similar to those that are obtained when the pump slips; hence, caution needs to be exercised when interpreting data of Ca(2+)-ATPase-mediated fluxes that display a non-linear dependence on the electrochemical proton (delta mu H) and/or calcium gradients (delta mu Ca). To address this issue, three experimentally verifiable relationships differentiating between membrane leak and enzymic slip were derived. First, by measuring delta mu H as a function of the rate of ATP hydrolysis by the enzyme. Second, by measuring the overall 'efficiency' of the pump as a function of delta mu H. Third, by measuring the proton ejection rate by the pump as a function of its ATP hydrolysis rate.

  13. Protein N-myristoylation in Escherichia coli: Reconstitution of a eukaryotic protein modification in bacteria

    International Nuclear Information System (INIS)

    Duronio, R.J.; Jackson-Machelski, E.; Heuckeroth, R.O.; Gordon, J.I.; Olins, P.O.; Devine, C.S.; Yonemoto, W.; Slice, L.W.; Taylor, S.S.

    1990-01-01

    Protein N-myristoylation refers to the covalent attachment of a myristoyl group (C14:0), via amide linkage, to the NH 2 -terminal glycine residue of certain cellular and viral proteins. Myristoyl-CoA:protein N-myristoyltransferase (NMT) catalyzes this cotranslational modification. The authors have developed a system for studying the substrate requirements and biological effects of protein N-myristoylation as well as NMT structure-activity relationships. Expression of the yeast NMT1 gene in Escherichia coli, a bacterium that has no endogenous NMT activity, results in production of the intact 53-kDa NMT polypeptide as well as a truncated polypeptide derived from proteolytic removal of its NH 2 -terminal 39 amino acids. By using a dual plasmid system, N-myristoylation of a mammalian protein was reconstituted in E. coli by simultaneous expression of the yeast NMT1 gene and a murine cDNA encoding the catalytic (C) subunit of cAMP-dependent protein kinase (PK-A). A major advantage of the bacterial system over eukaryotic systems is the absence of endogenous NMT and substrates, providing a more straightforward way of preparing myristoylated, analog-substituted, and nonmyristoylated forms of a given protein for comparison of their structural and functional properties. The experimental system may prove useful for recapitulating other eukaryotic protein modifications in E. coli so that structure-activity relationships of modifying enzymes and their substrates can be more readily assessed

  14. Late onset of cryptococcal cervical lymphadenitis following immune reconstitution inflammatory syndrome in a patient with AIDS.

    Science.gov (United States)

    Sethupathi, Meenakshi; Yoganathan, Kathir

    2015-01-06

    A 32-year-old woman was diagnosed HIV positive with disseminated cryptococcal infection in May 2006. Her initial CD4 was 7 cells/µL and she had a right supraclavicular nodal mass, which was biopsied and shown to be consistent with cryptococcal lymphadenitis. She was treated for disseminated cryptococcal infection and was started on antiretroviral medications subsequently. Two years later, she developed a left supraclavicular mass. Her CD4 count was 320 cells/µL and HIV RNA level was undetectable. Investigations and biopsy results were consistent with a late presentation of cryptococcal immune reconstitution inflammatory syndrome (IRIS). She was treated with oral corticosteroids and her symptoms resolved completely. IRIS is a recognised complication of HIV treatment and occurs in a significant percentage of patients within the first 3 months of starting antiretroviral therapy. This case report illustrates the importance of recognising late presentations of IRIS. It is vital to differentiate true cryptococcal lymphadenitis from IRIS-induced cryptococcal lymphadenitis. 2015 BMJ Publishing Group Ltd.

  15. A statin-loaded reconstituted high-density lipoprotein nanoparticle inhibits atherosclerotic plaque inflammation

    Science.gov (United States)

    Duivenvoorden, Raphaël; Tang, Jun; Cormode, David P.; Mieszawska, Aneta J.; Izquierdo-Garcia, David; Ozcan, Canturk; Otten, Maarten J.; Zaidi, Neeha; Lobatto, Mark E.; van Rijs, Sarian M.; Priem, Bram; Kuan, Emma L.; Martel, Catherine; Hewing, Bernd; Sager, Hendrik; Nahrendorf, Matthias; Randolph, Gwendalyn J.; Stroes, Erik S. G.; Fuster, Valentin; Fisher, Edward A.; Fayad, Zahi A.; Mulder, Willem J. M.

    2014-01-01

    Inflammation is a key feature of atherosclerosis and a target for therapy. Statins have potent anti-inflammatory properties but these cannot be fully exploited with oral statin therapy due to low systemic bioavailability. Here we present an injectable reconstituted high-density lipoprotein (rHDL) nanoparticle carrier vehicle that delivers statins to atherosclerotic plaques. We demonstrate the anti-inflammatory effect of statin-rHDL in vitro and show that this effect is mediated through the inhibition of the mevalonate pathway. We also apply statin-rHDL nanoparticles in vivo in an apolipoprotein E-knockout mouse model of atherosclerosis and show that they accumulate in atherosclerotic lesions in which they directly affect plaque macrophages. Finally, we demonstrate that a 3-month low-dose statin-rHDL treatment regimen inhibits plaque inflammation progression, while a 1-week high-dose regimen markedly decreases inflammation in advanced atherosclerotic plaques. Statin-rHDL represents a novel potent atherosclerosis nanotherapy that directly affects plaque inflammation.

  16. Effects of benzo[a]pyrene-DNA adducts on a reconstituted replication system

    International Nuclear Information System (INIS)

    Brown, W.C.; Romano, L.J.

    1991-01-01

    The authors have used a partially reconstituted replication system consisting of T7 DNA polymerase and T7 gene 4 protein to examine the effect of benzo[a]pyrene (B[a]P) adducts on DNA synthesis and gene 4 protein activities. The gene 4 protein is required for T7 DNA replication because of its ability to act as both a primase and helicase. They show here that total synthesis decreases as the level of adducts per molecule of DNA increases, suggesting that the B[a]P adducts are blocking an aspect of the replication process. By challenging synthesis on oligonucleotide-primed B[a]P-modified DNA with unmodified DNA, they present evidence that the T7 DNA polymerase freely dissociates after encountering an adduct. Prior studies have shown that the gene 4 protein alone does not dissociate from the template during translocation upon encountering an adduct. However, when gene 4 protein primed DNA synthesis is challenged, they observe an increase in synthesis but to a lesser extent than observed on oligonucleotide-primed synthesis. Finally, they have examined DNA synthesis on duplex templates and show the B[a]P adducts inhibit synthesis by the T7 DNA polymerase and gene 4 protein to the same extent regardless of whether the adducts are positioned in the leading or lagging strand, while synthesis by the polymerase alone is inhibited only when the adducts are in the template strand

  17. Is vitamin D deficiency involved in the immune reconstitution inflammatory syndrome?

    Directory of Open Access Journals (Sweden)

    Moreno-Reyes Rodrigo

    2009-04-01

    Full Text Available Abstract Background About 20–30% of persons with HIV infection, especially those living in countries with limited resources, experience an immune reconstitution inflammatory syndrome (IRIS after starting antiretroviral treatment. The active form of vitamin D, 1,25-dihydroxyvitamin D, is a key player in the clearance of pathogens and influences the level of inflammation and macrophage activation. Presentation of the hypothesis We hypothesize that low availability of 1,25-dihydroxyvitamin D, either due to vitamin D deficiency or due to polymorphisms in the vitamin D receptor or in its activating/inactivating enzymes, contributes to the appearance of IRIS. Furthermore, drug interactions with the enzymatic pathways of vitamin D could favour the development of IRIS. Testing the hypothesis Our hypothesis could be explored by a case-control study to assess the prevalence of vitamin D deficiency in HIV-infected patients on antiretroviral treatment who develop and do not develop IRIS. Implications of the hypothesis If the role of vitamin D in IRIS is confirmed, we would be able to screen patients at risk for IRIS by screening for vitamin D deficiency. After confirmation by means of a clinical trial, vitamin D supplementation could be a cheap and safe way to reduce the incidence of IRIS.

  18. Active transport of Na/sup +/ by reconstituted Na,K-ATPase

    Energy Technology Data Exchange (ETDEWEB)

    Boldyrev, A.A.; Svinukhova, I.A.

    1987-02-20

    The ability of ATP, CTP, ITP, GTP, and UTP to support ouabain-sensitive accumulation of Na/sup +/ by proteoliposomes with a reconstituted Na/K-pump was investigated. At a low (Na/sup +/)/(K/sup +/) ratio in the medium (20 mM/50 mM), a correlation is observed between the proton-accepting capacity of the nucleotide and its effectiveness as a substrate of active transport. To test the hypothesis of the importance of the presence of a negative charge in the 1-position of the purine (3-pyrimidine) base of the nucleotide for mutual transitions between the Na- and K-conformations of Na,K-ATPase they used two analogs of ATP: N/sub 1/-hydroxy-ATP, possessing proton acceptor capacity, and N/sub 1/-methoxy-ATP, in the molecule of which the negative charge is quenched by a methyl group. The first substrate supports active accumulation of Na/sup +/ in proteoliposomes at the same rate as ATP, whereas the second substrate is relatively ineffective.

  19. Endothelial jagged-2 sustains hematopoietic stem and progenitor reconstitution after myelosuppression.

    Science.gov (United States)

    Guo, Peipei; Poulos, Michael G; Palikuqi, Brisa; Badwe, Chaitanya R; Lis, Raphael; Kunar, Balvir; Ding, Bi-Sen; Rabbany, Sina Y; Shido, Koji; Butler, Jason M; Rafii, Shahin

    2017-12-01

    Angiocrine factors, such as Notch ligands, supplied by the specialized endothelial cells (ECs) within the bone marrow and splenic vascular niche play an essential role in modulating the physiology of adult hematopoietic stem and progenitor cells (HSPCs). However, the relative contribution of various Notch ligands, specifically jagged-2, to the homeostasis of HSPCs is unknown. Here, we show that under steady state, jagged-2 is differentially expressed in tissue-specific vascular beds, but its expression is induced in hematopoietic vascular niches after myelosuppressive injury. We used mice with EC-specific deletion of the gene encoding jagged-2 (Jag2) to demonstrate that while EC-derived jagged-2 was dispensable for maintaining the capacity of HSPCs to repopulate under steady-state conditions, by activating Notch2 it did contribute to the recovery of HSPCs in response to myelosuppressive conditions. Engraftment and/or expansion of HSPCs was dependent on the expression of endothelial-derived jagged-2 following myeloablation. Additionally, jagged-2 expressed in bone marrow ECs regulated HSPC cell cycle and quiescence during regeneration. Endothelial-deployed jagged-2 triggered Notch2/Hey1, while tempering Notch2/Hes1 signaling in HSPCs. Collectively, these data demonstrate that EC-derived jagged-2 activates Notch2 signaling in HSPCs to promote hematopoietic recovery and has potential as a therapeutic target to accelerate balanced hematopoietic reconstitution after myelosuppression.

  20. Reconstitution of a eukaryotic replisome reveals suppression mechanisms that define leading/lagging strand operation

    Science.gov (United States)

    Georgescu, Roxana E; Schauer, Grant D; Yao, Nina Y; Langston, Lance D; Yurieva, Olga; Zhang, Dan; Finkelstein, Jeff; O'Donnell, Mike E

    2015-01-01

    We have reconstituted a eukaryotic leading/lagging strand replisome comprising 31 distinct polypeptides. This study identifies a process unprecedented in bacterial replisomes. While bacteria and phage simply recruit polymerases to the fork, we find that suppression mechanisms are used to position the distinct eukaryotic polymerases on their respective strands. Hence, Pol ε is active with CMG on the leading strand, but it is unable to function on the lagging strand, even when Pol δ is not present. Conversely, Pol δ-PCNA is the only enzyme capable of extending Okazaki fragments in the presence of Pols ε and α. We have shown earlier that Pol δ-PCNA is suppressed on the leading strand with CMG (Georgescu et al., 2014). We propose that CMG, the 11-subunit helicase, is responsible for one or both of these suppression mechanisms that spatially control polymerase occupancy at the fork. DOI: http://dx.doi.org/10.7554/eLife.04988.001 PMID:25871847

  1. Native and reconstituted HDL activate Stat3 in ventricular cardiomyocytes via ERK1/2: role of sphingosine-1-phosphate.

    Science.gov (United States)

    Frias, Miguel A; James, Richard W; Gerber-Wicht, Christine; Lang, Ursula

    2009-05-01

    High-density lipoprotein (HDL) has been reported to have cardioprotective properties independent from its cholesterol transport activity. The influence of native HDL and reconstituted HDL (rHDL) on Stat3, the transcription factor playing an important role in myocardium adaptation to stress, was analysed in neonatal rat ventricular cardiomyocytes. We have investigated modulating the composition of rHDL as a means of expanding its function and potential cardioprotective effects. Stat3 phosphorylation and activation were determined by western blotting and electrophoretic mobility shift assay (EMSA). In ventricular cardiomyocytes, HDL and the HDL constituent sphingosine-1-phosphate (S1P) induce a concentration- and time-dependent increase in Stat3 activation. They also enhance extracellular signal-regulated kinases (ERK1/2) and p38 mitogen-activated protein kinase (MAPK) phosphorylation. U0126, a specific inhibitor of MEK1/2, the upstream activator of ERK1/2, abolishes HDL- and S1P-induced Stat3 activation, whereas the p38 MAPK blocker SB203580 has no significant effect. Inhibition of the tyrosine kinase family Src (Src) caused a significant reduction of Stat3 activation, whereas inhibition of phosphatidylinositol 3-kinase (PI3K) had no effect. S1P and rHDL containing S1P have a similar strong stimulatory action on Stat3, ERK1/2, and p38 MAPK comparable to native HDL. S1P-free rHDL has a much weaker effect. Experiments with agonists and antagonists of the S1P receptor subtypes indicate that HDL and S1P activate Stat3 mainly through the S1P2 receptor. In ventricular cardiomyocytes, addition of S1P to rHDL enhances its therapeutic potential by improving its capacity to activate Stat3. Activation of Stat3 occurs mainly via the S1P constituent and the lipid receptor S1P2 requiring stimulation of ERK1/2 and Src but not p38 MAPK or PI3K. The study underlines the therapeutic potential of tailoring rHDL to confront particular clinical situations.

  2. Evaluation of cellular phenotypes implicated in immunopathogenesis and monitoring immune reconstitution inflammatory syndrome in HIV/leprosy cases.

    Science.gov (United States)

    Giacoia-Gripp, Carmem Beatriz Wagner; Sales, Anna Maria; Nery, José Augusto da Costa; Santos-Oliveira, Joanna Reis; de Oliveira, Ariane Leite; Sarno, Euzenir Nunes; Morgado, Mariza Gonçalves

    2011-01-01

    It is now evident that HAART-associated immunological improvement often leads to a variety of new clinical manifestations, collectively termed immune reconstitution inflammatory syndrome, or IRIS. This phenomenon has already been described in cases of HIV coinfection with Mycobacterium leprae, most of them belonging to the tuberculoid spectrum of leprosy disease, as observed in leprosy reversal reaction (RR). However, the events related to the pathogenesis of this association need to be clarified. This study investigated the immunological profile of HIV/leprosy patients, with special attention to the cellular activation status, to better understand the mechanisms related to IRIS/RR immunopathogenesis, identifying any potential biomarkers for IRIS/RR intercurrence. Eighty-five individuals were assessed in this study: HIV/leprosy and HIV-monoinfected patients, grouped according to HIV-viral load levels, leprosy patients without HIV coinfection, and healthy controls. Phenotypes were evaluated by flow cytometry for T cell subsets and immune differentiation/activation markers. As expected, absolute counts of the CD4+ and CD8+ T cells from the HIV-infected individuals changed in relation to those of the leprosy patients and controls. However, there were no significant differences among the groups, whether in the expression of cellular differentiation phenotypes or cellular activation, as reflected by the expression of CD38 and HLA-DR. Six HIV/leprosy patients identified as IRIS/RR were analyzed during IRIS/RR episodes and after prednisone treatment. These patients presented high cellular activation levels regarding the expression of CD38 in CD8+ cells T during IRIS/RR (median: 77,15%), dropping significantly (p<0,05) during post-IRIS/RR moments (median: 29,7%). Furthermore, an increase of cellular activation seems to occur prior to IRIS/RR. These data suggest CD38 expression in CD8+ T cells interesting tool identifying HIV/leprosy individuals at risk for IRIS/RR. So, a

  3. Evaluation of cellular phenotypes implicated in immunopathogenesis and monitoring immune reconstitution inflammatory syndrome in HIV/leprosy cases.

    Directory of Open Access Journals (Sweden)

    Carmem Beatriz Wagner Giacoia-Gripp

    Full Text Available BACKGROUND: It is now evident that HAART-associated immunological improvement often leads to a variety of new clinical manifestations, collectively termed immune reconstitution inflammatory syndrome, or IRIS. This phenomenon has already been described in cases of HIV coinfection with Mycobacterium leprae, most of them belonging to the tuberculoid spectrum of leprosy disease, as observed in leprosy reversal reaction (RR. However, the events related to the pathogenesis of this association need to be clarified. This study investigated the immunological profile of HIV/leprosy patients, with special attention to the cellular activation status, to better understand the mechanisms related to IRIS/RR immunopathogenesis, identifying any potential biomarkers for IRIS/RR intercurrence. METHODS/PRINCIPAL FINDINGS: Eighty-five individuals were assessed in this study: HIV/leprosy and HIV-monoinfected patients, grouped according to HIV-viral load levels, leprosy patients without HIV coinfection, and healthy controls. Phenotypes were evaluated by flow cytometry for T cell subsets and immune differentiation/activation markers. As expected, absolute counts of the CD4+ and CD8+ T cells from the HIV-infected individuals changed in relation to those of the leprosy patients and controls. However, there were no significant differences among the groups, whether in the expression of cellular differentiation phenotypes or cellular activation, as reflected by the expression of CD38 and HLA-DR. Six HIV/leprosy patients identified as IRIS/RR were analyzed during IRIS/RR episodes and after prednisone treatment. These patients presented high cellular activation levels regarding the expression of CD38 in CD8+ cells T during IRIS/RR (median: 77,15%, dropping significantly (p<0,05 during post-IRIS/RR moments (median: 29,7%. Furthermore, an increase of cellular activation seems to occur prior to IRIS/RR. CONCLUSION/SIGNIFICANCE: These data suggest CD38 expression in CD8+ T cells

  4. OGLE-2017-BLG-0329L: A Microlensing Binary Characterized with Dramatically Enhanced Precision Using Data from Space-based Observations

    Science.gov (United States)

    Han, C.; Calchi Novati, S.; Udalski, A.; Lee, C.-U.; Gould, A.; Bozza, V.; Mróz, P.; Pietrukowicz, P.; Skowron, J.; Szymański, M. K.; Poleski, R.; Soszyński, I.; Kozłowski, S.; Ulaczyk, K.; Pawlak, M.; Rybicki, K.; Iwanek, P.; The OGLE Collaboration; Albrow, M. D.; Chung, S.-J.; Hwang, K.-H.; Jung, Y. K.; Ryu, Y.-H.; Shin, I.-G.; Shvartzvald, Y.; Yee, J. C.; Zang, W.; Zhu, W.; Cha, S.-M.; Kim, D.-J.; Kim, H.-W.; Kim, S.-L.; Lee, D.-J.; Lee, Y.; Park, B.-G.; Pogge, R. W.; Kim, W.-T.; The KMTNet Collaboration; Beichman, C.; Bryden, G.; Carey, S.; Gaudi, B. S.; Henderson, C. B.; The Spitzer Team; Dominik, M.; Helling, C.; Hundertmark, M.; Jørgensen, U. G.; Longa-Peña, P.; Lowry, S.; Sajadian, S.; Burgdorf, M. J.; Campbell-White, J.; Ciceri, S.; Evans, D. F.; Haikala, L. K.; Hinse, T. C.; Rahvar, S.; Rabus, M.; Snodgrass, C.; The MiNDSTEp Collaboration

    2018-06-01

    Mass measurements of gravitational microlenses require one to determine the microlens parallax π E, but precise π E measurement, in many cases, is hampered due to the subtlety of the microlens-parallax signal combined with the difficulty of distinguishing the signal from those induced by other higher-order effects. In this work, we present the analysis of the binary-lens event OGLE-2017-BLG-0329, for which π E is measured with a dramatically improved precision using additional data from space-based Spitzer observations. We find that while the parallax model based on the ground-based data cannot be distinguished from a zero-π E model at the 2σ level, the addition of the Spitzer data enables us to identify two classes of solutions, each composed of a pair of solutions according to the well-known ecliptic degeneracy. It is found that the space-based data reduce the measurement uncertainties of the north and east components of the microlens-parallax vector {{\\boldsymbol{π }}}{{E}} by factors ∼18 and ∼4, respectively. With the measured microlens parallax combined with the angular Einstein radius measured from the resolved caustic crossings, we find that the lens is composed of a binary with component masses of either (M 1, M 2) ∼ (1.1, 0.8) M ⊙ or ∼(0.4, 0.3) M ⊙ according to the two solution classes. The first solution is significantly favored but the second cannot be securely ruled out based on the microlensing data alone. However, the degeneracy can be resolved from adaptive optics observations taken ∼10 years after the event.

  5. Homogeneous immunoglobulins in the serum of irradiated and bone marrow reconstituted mice: the role of thymus and spleen

    International Nuclear Information System (INIS)

    Mink, J.G.; Radl, J.; Berg, P. van den; Muiswinkel, W.B. van; Oosterom, R. van.

    1979-01-01

    The influence of thymectomy and splenectomy on the frequency and class distribution of homogeneous immunoglobulins (H-Ig) in serum was studied in lethally irradiated (DBA/2 x C57B1/Rij)F 1 mice reconstituted with syngeneic bone marrow. During four follow-up periods in the first 9 months after transplantation, the sham-operated controls and splenectomized animals developed transient H-Ig in an average frequency of 14.2 and 15.7% respectively. There were no marked differences in the incidence of H-Ig within these two groups. In contrast, thymectomized mice and mice both thymectomized and splenectomized showed H-Ig in much higher frequencies (average percentages 31.6 and 36.5 respectively). The highest frequency of H-Ig was observed between 1.5 and 3.5 months after transplantation. H-Ig of the IgG1 and IgG2 subclasses were most frequent in all groups during the first 3.5 months. Later, H-Ig belonging to the IgM class also appeared in somewhat higher numbers. H-Ig of the IgA class was a very rare finding at any time. These results indicate that the presence of the thymus, but not necessarily of the spleen, is an important factor in the regulation of the immunoglobulin heterogeneity during the reconstitution of the immune system in lethally irradiated and bone marrow reconstituted mice. (author)

  6. Evaluating Complex Mixtures in the Zebrafish Embryo by Reconstituting Field Water Samples: A Metal Pollution Case Study

    Directory of Open Access Journals (Sweden)

    Ellen D. G. Michiels

    2017-03-01

    Full Text Available Accurately assessing the toxicity of complex, environmentally relevant mixtures remains an important challenge in ecotoxicology. The goal was to identify biological effects after exposure to environmental water samples and to determine whether the observed effects could be explained by the waterborne metal mixture found in the samples. Zebrafish embryos were exposed to water samples of five different sites originating from two Flemish (Mol and Olen, Belgium metal contaminated streams: “Scheppelijke Nete” (SN and “Kneutersloop” (K, and a ditch (D, which is the contamination source of SN. Trace metal concentrations, and Na, K, Mg and Ca concentrations were measured using ICP-MS and were used to reconstitute site-specific water samples. We assessed whether the effects that were observed after exposure to environmental samples could be explained by metal mixture toxicity under standardized laboratory conditions. Exposure to “D” or “reconstituted D” water caused 100% mortality. SN and reconstituted SN water caused similar effects on hatching, swim bladder inflation, growth and swimming activity. A canonical discriminant analysis confirmed a high similarity between both exposure scenarios, indicating that the observed toxicity was indeed primarily caused by metals. The applied workflow could be a valuable approach to evaluate mixture toxicity that limits time and costs while maintaining biological relevance.

  7. Studies on cell-free metabolism: ethanol production by a yeast glycolytic system reconstituted from purified enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Welch, P; Scopes, R K

    1985-07-01

    A reconstituted glycolytic system has been established from individually purified enzymes to simulate the conversion of glucose to ethanol plus CO/sub 2/ by yeast. Sustained and extensive conversion occurred provided that input of glucose matched the rate of ATP degradation appropriately. ATPase activity could be replaced by arsenate, which uncoupled ATP synthesis from glycolysis. The mode of uncoupling was investigated, and it was concluded that the artificial intermediate, 1-arseno-3-phosphoglycerate, has a half-life of no more than a few milliseconds. Arsenate at 4 mM concentration could simulate the equivalent of 10 ..mu..mol/ml min. of ATPase activity. The reconstituted enzyme system was capable of totally degrading one M (18% w/v) glucose in 8 hours giving 9% (w/v) ethanol. The levels of metabolites during metabolism were measured to detect rate-limiting steps. The successful operation of the reconstituted enzyme system demonstrates that it is possible to carry out complex chemical transformations with multiple enzyme systems in vitro. 36 references.

  8. Fabrication and characterization of an integrated ionic device from suspended polypyrrole and alamethicin-reconstituted lipid bilayer membranes

    International Nuclear Information System (INIS)

    Northcutt, Robert; Sundaresan, Vishnu-Baba

    2012-01-01

    Conducting polymers are electroactive materials that undergo conformal relaxation of the polymer backbone in the presence of an electrical field through ion exchange with solid or aqueous electrolytes. This conformal relaxation and the associated morphological changes make conducting polymers highly suitable for actuation and sensing applications. Among smart materials, bioderived active materials also use ion transport for sensing and actuation functions via selective ion transport. The transporter proteins extracted from biological cell membranes and reconstituted into a bilayer lipid membrane in bioderived active materials regulate ion transport for engineering functions. The protein transporter reconstituted in the bilayer lipid membrane is referred to as the bioderived membrane and serves as the active component in bioderived active materials. Inspired by the similarities in the physics of transduction in conducting polymers and bioderived active materials, an integrated ionic device is formed from the bioderived membrane and the conducting polymer membrane. This ionic device is fabricated into a laminated thin-film membrane and a common ion that can be processed by the bioderived and the conducting polymer membranes couple the ionic function of these two membranes. An integrated ionic device, fabricated from polypyrrole (PPy) doped with sodium dodecylbenzenesulfonate (NaDBS) and an alamethicin-reconstituted DPhPC bilayer lipid membrane, is presented in this paper. A voltage-gated sodium current regulates the electrochemical response in the PPy(DBS) layer. The integrated device is fabricated on silicon-based substrates through microfabrication, electropolymerization, and vesicle fusion, and ionic activity is characterized through electrochemical measurements. (paper)

  9. Perceived Parental Acceptance Related to Self-Esteem, GPA, Sex-Role Identity, and Substance Use of Adolescents From Intact and Reconstituted Families

    OpenAIRE

    Sniteman, Stephen B.

    1993-01-01

    This investigation assessed the relationship between adolescents of intact families and adolescents in reconstituted families with regard to the effects of perception of parental acceptance on the variables of self-esteem, academic performance, sex role identity, and use o f substances. Observed differences between adolescents of intact and reconstituted families from a structural perspective, eliminating process variables, were also examined. Participants included two hundred fifty-six high ...

  10. Keratinocytes propagated in serum-free, feeder-free culture conditions fail to form stratified epidermis in a reconstituted skin model.

    Directory of Open Access Journals (Sweden)

    Rebecca Lamb

    Full Text Available Primary human epidermal stem cells isolated from skin tissues and subsequently expanded in tissue culture are used for human therapeutic use to reconstitute skin on patients and to generate artificial skin in culture for academic and commercial research. Classically, epidermal cells, known as keratinocytes, required fibroblast feeder support and serum-containing media for serial propagation. In alignment with global efforts to remove potential animal contaminants, many serum-free, feeder-free culture methods have been developed that support derivation and growth of these cells in 2-dimensional culture. Here we show that keratinocytes grown continually in serum-free and feeder-free conditions were unable to form into a stratified, mature epidermis in a skin equivalent model. This is not due to loss of cell potential as keratinocytes propagated in serum-free, feeder-free conditions retain their ability to form stratified epidermis when re-introduced to classic serum-containing media. Extracellular calcium supplementation failed to improve epidermis development. In contrast, the addition of serum to commercial, growth media developed for serum-free expansion of keratinocytes facilitated 3-dimensional stratification in our skin equivalent model. Moreover, the addition of heat-inactivated serum improved the epidermis structure and thickness, suggesting that serum contains factors that both aid and inhibit stratification.

  11. Production of Volatile Compounds in Reconstituted Milk Reduced-Fat Cheese and the Physicochemical Properties as Affected by Exopolysaccharide-Producing Strain

    Directory of Open Access Journals (Sweden)

    Weijun Wang

    2012-12-01

    Full Text Available The application of the exopolysaccharide-producing strains for improving the texture and technical properties of reduced-fat cheese looks very promising. Streptococcus thermophilus TM11 was evaluated for production of reduced-fat cheese using reconstituted milk powder (CRMP. The physicochemical analysis of fresh and stored cheeses showed that this strain slightly increased moisture content resulting in cheese with higher yield and lower protein content compared to the direct acidified cheese. The volatiles of cheese were determined by SPME and GC equipped with a mass spectrometer. The results indicated that the major compounds included aldehydes, ketones and acids, whereas, alcohols and branched-chain aldehydes that contribute to exciting and harsh flavors were not found in CRMP. By the textural profile analysis, we found the cheese made with S. thermophilus TM11 had lower cohesiveness, resilience and higher adhesiveness than the direct acidified cheese, and had similar hardness. Further, S. thermophilus TM11 greatly changed the protein matrix with more opened cavities according to observation by scanning electron microscopy. Consequently, use of S. thermophilus TM11 could endow CRMP with the novel and suitable flavor properties and improved texture quality.

  12. Molecular pathways undergoing dramatic transcriptomic changes during tumor development in the human colon

    Directory of Open Access Journals (Sweden)

    Maglietta Rosalia

    2012-12-01

    Full Text Available Abstract Background The malignant transformation of precancerous colorectal lesions involves progressive alterations at both the molecular and morphologic levels, the latter consisting of increases in size and in the degree of cellular atypia. Analyzing preinvasive tumors of different sizes can therefore shed light on the sequence of these alterations. Methods We used a molecular pathway-based approach to analyze transcriptomic profiles of 59 colorectal tumors representing early and late preinvasive stages and the invasive stage of tumorigenesis. Random set analysis was used to identify biological pathways enriched for genes differentially regulated in tumors (compared with 59 samples of normal mucosa. Results Of the 880 canonical pathways we investigated, 112 displayed significant tumor-related upregulation or downregulation at one or more stages of tumorigenesis. This allowed us to distinguish between pathways whose dysregulation is probably necessary throughout tumorigenesis and those whose involvement specifically drives progression from one stage to the next. We were also able to pinpoint specific changes within each gene set that seem to play key roles at each transition. The early preinvasive stage was characterized by cell-cycle checkpoint activation triggered by DNA replication stress and dramatic downregulation of basic transmembrane signaling processes that maintain epithelial/stromal homeostasis in the normal mucosa. In late preinvasive lesions, there was also downregulation of signal transduction pathways (e.g., those mediated by G proteins and nuclear hormone receptors involved in cell differentiation and upregulation of pathways governing nuclear envelope dynamics and the G2>M transition in the cell cycle. The main features of the invasive stage were activation of the G1>S transition in the cell cycle, upregulated expression of tumor-promoting microenvironmental factors, and profound dysregulation of metabolic pathways (e

  13. The Dramatic Size and Kinematic Evolution of Massive Early-type Galaxies

    Science.gov (United States)

    Lapi, A.; Pantoni, L.; Zanisi, L.; Shi, J.; Mancuso, C.; Massardi, M.; Shankar, F.; Bressan, A.; Danese, L.

    2018-04-01

    We aim to provide a holistic view on the typical size and kinematic evolution of massive early-type galaxies (ETGs) that encompasses their high-z star-forming progenitors, their high-z quiescent counterparts, and their configurations in the local Universe. Our investigation covers the main processes playing a relevant role in the cosmic evolution of ETGs. Specifically, their early fast evolution comprises biased collapse of the low angular momentum gaseous baryons located in the inner regions of the host dark matter halo; cooling, fragmentation, and infall of the gas down to the radius set by the centrifugal barrier; further rapid compaction via clump/gas migration toward the galaxy center, where strong heavily dust-enshrouded star formation takes place and most of the stellar mass is accumulated; and ejection of substantial gas amount from the inner regions by feedback processes, which causes a dramatic puffing-up of the stellar component. In the late slow evolution, passive aging of stellar populations and mass additions by dry merger events occur. We describe these processes relying on prescriptions inspired by basic physical arguments and by numerical simulations to derive new analytical estimates of the relevant sizes, timescales, and kinematic properties for individual galaxies along their evolution. Then we obtain quantitative results as a function of galaxy mass and redshift, and compare them to recent observational constraints on half-light size R e , on the ratio v/σ between rotation velocity and velocity dispersion (for gas and stars) and on the specific angular momentum j ⋆ of the stellar component; we find good consistency with the available multiband data in average values and dispersion, both for local ETGs and for their z ∼ 1–2 star-forming and quiescent progenitors. The outcomes of our analysis can provide hints to gauge sub-grid recipes implemented in simulations, to tune numerical experiments focused on specific processes, and to plan

  14. Engineered split in Pfu DNA polymerase fingers domain improves incorporation of nucleotide γ-phosphate derivative

    Science.gov (United States)

    Hansen, Connie J.; Wu, Lydia; Fox, Jeffrey D.; Arezi, Bahram; Hogrefe, Holly H.

    2011-01-01

    Using compartmentalized self-replication (CSR), we evolved a version of Pyrococcus furiosus (Pfu) DNA polymerase that tolerates modification of the γ-phosphate of an incoming nucleotide. A Q484R mutation in α-helix P of the fingers domain, coupled with an unintended translational termination-reinitiation (split) near the finger tip, dramatically improve incorporation of a bulky γ-phosphate-O-linker-dabcyl substituent. Whether synthesized by coupled translation from a bicistronic (−1 frameshift) clone, or reconstituted from separately expressed and purified fragments, split Pfu mutant behaves identically to wild-type DNA polymerase with respect to chromatographic behavior, steady-state kinetic parameters (for dCTP), and PCR performance. Although naturally-occurring splits have been identified previously in the finger tip region of T4 gp43 variants, this is the first time a split (in combination with a point mutation) has been shown to broaden substrate utilization. Moreover, this latest example of a split hyperthermophilic archaeal DNA polymerase further illustrates the modular nature of the Family B DNA polymerase structure. PMID:21062827

  15. Engineered split in Pfu DNA polymerase fingers domain improves incorporation of nucleotide gamma-phosphate derivative.

    Science.gov (United States)

    Hansen, Connie J; Wu, Lydia; Fox, Jeffrey D; Arezi, Bahram; Hogrefe, Holly H

    2011-03-01

    Using compartmentalized self-replication (CSR), we evolved a version of Pyrococcus furiosus (Pfu) DNA polymerase that tolerates modification of the γ-phosphate of an incoming nucleotide. A Q484R mutation in α-helix P of the fingers domain, coupled with an unintended translational termination-reinitiation (split) near the finger tip, dramatically improve incorporation of a bulky γ-phosphate-O-linker-dabcyl substituent. Whether synthesized by coupled translation from a bicistronic (-1 frameshift) clone, or reconstituted from separately expressed and purified fragments, split Pfu mutant behaves identically to wild-type DNA polymerase with respect to chromatographic behavior, steady-state kinetic parameters (for dCTP), and PCR performance. Although naturally-occurring splits have been identified previously in the finger tip region of T4 gp43 variants, this is the first time a split (in combination with a point mutation) has been shown to broaden substrate utilization. Moreover, this latest example of a split hyperthermophilic archaeal DNA polymerase further illustrates the modular nature of the Family B DNA polymerase structure.

  16. Significant effect of Ca2+ on improving the heat resistance of lactic acid bacteria.

    Science.gov (United States)

    Huang, Song; Chen, Xiao Dong

    2013-07-01

    The heat resistance of lactic acid bacteria (LAB) has been extensively investigated due to its highly practical significance. Reconstituted skim milk (RSM) has been found to be one of the most effective protectant wall materials for microencapsulating microorganisms during convective drying, such as spray drying. In addition to proteins and carbohydrate, RSM is rich in calcium. It is not clear which component is critical in the RSM protection mechanism. This study investigated the independent effect of calcium. Ca(2+) was added to lactose solution to examine its influence on the heat resistance of Lactobacillus rhamnosus ZY, Lactobacillus casei Zhang, Lactobacillus plantarum P8 and Streptococcus thermophilus ND03. The results showed that certain Ca(2+) concentrations enhanced the heat resistance of the LAB strains to different extents, that is produced higher survival and shorter regrowth lag times of the bacterial cells. In some cases, the improvements were dramatic. More scientifically insightful and more intensive instrumental study of the Ca(2+) behavior around and in the cells should be carried out in the near future. In the meantime, this work may lead to the development of more cost-effective wall materials with Ca(2+) added as a prime factor. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  17. Intravenous application of an anticalin dramatically lowers plasma digoxin levels and reduces its toxic effects in rats

    Energy Technology Data Exchange (ETDEWEB)

    Eyer, Florian, E-mail: Florian.Eyer@mac.com [Department of Toxicology, Klinikum rechts der Isar, Munich (Germany); Steimer, Werner [Institute of Clinical Chemistry and Pathobiochemistry, Klinikum rechts der Isar, Munich (Germany); Nitzsche, Thomas [Munich Center for Integrated Protein Science (CIPS-M), Freising-Weihenstephan (Germany); Lehrstuhl für Biologische Chemie, Technische Universität München, Freising-Weihenstephan (Germany); Jung, Nicole; Neuberger, Heidi [Department of Toxicology, Klinikum rechts der Isar, Munich (Germany); Müller, Christine [Institute of Clinical Chemistry and Pathobiochemistry, Klinikum rechts der Isar, Munich (Germany); Schlapschy, Martin [Munich Center for Integrated Protein Science (CIPS-M), Freising-Weihenstephan (Germany); Lehrstuhl für Biologische Chemie, Technische Universität München, Freising-Weihenstephan (Germany); Zilker, Thomas [Department of Toxicology, Klinikum rechts der Isar, Munich (Germany); Skerra, Arne [Munich Center for Integrated Protein Science (CIPS-M), Freising-Weihenstephan (Germany); Lehrstuhl für Biologische Chemie, Technische Universität München, Freising-Weihenstephan (Germany)

    2012-09-15

    Lipocalins tailored with high affinity for prescribed ligands, so-called anticalins, constitute promising candidates as antidotes. Here, we present an animal study to investigate both pharmacokinetic and clinical effects of an anticalin specific for the digitalis compound digoxin. Intravenous digoxin (2.5–50 μg/kg/min) was administered to rats until first changes in the ECG occurred (dose finding study) or a priori for 30 min (kinetic study). The anticalin DigA16(H86N), dubbed DigiCal, was administered intravenously at absolute doses of 1, 5, 10 and 20 mg, while the control group received isotonic saline. Hemodynamic changes, several ECG parameters and digoxin concentration in plasma were monitored at given time intervals. After DigiCal administration free digoxin concentration in plasma ultrafiltrate declined dramatically within 1 min to the presumably non-toxic range. There was also a significant and DigiCal dose-dependent effect on longer survival, less ECG alterations, arrhythmia, and improved hemodynamics. Infusion of a lower digoxin dose (2.5 μg/kg/min) resulted in a more sustained reduction of free digoxin in plasma after DigiCal administration compared to a higher digoxin dose (25 μg/kg/min), whereas ECG and hemodynamic parameters did not markedly differ, reflecting the known relative insensitivity of rats towards digoxin toxicity. Notably, we observed a re-increase of free digoxin in plasma some time after bolus administration of DigiCal, which was presumably due to toxin redistribution from tissue in combination with the relatively fast renal clearance of the rather small protein antidote. We conclude that anticalins with appropriately engineered drug-binding activities and, possibly, prolonged plasma half-life offer prospects for next-generation antidotal therapy. -- Highlights: ► We provide an advanced model of digoxin toxicity in rats. ► We report on binding of digoxin to a novel designed anticalin. ► We report on pharmacokinetics of digoxin

  18. BRAF-V600E mutant papillary craniopharyngioma dramatically responds to combination BRAF and MEK inhibitors.

    Science.gov (United States)

    Roque, Ashley; Odia, Yazmin

    2017-04-01

    We present a patient with BRAF-V600E mutant papillary craniopharyngioma successfully treated with combination BRAF (dabrafenib 150 mg twice daily) and MEK (trametinib 2 mg daily) inhibitors after her unresectable tumor proved refractory to radiation. Serial brain MRIs and PET revealed marked tumor reduction with gradual neurological improvement and permanent panhypopituitarism.

  19. Learning to Lead against the Grain: Dramatizing the Emotional Toll of Teacher Leadership

    Science.gov (United States)

    Cranston, Jerome; Kusanovich, Kristin

    2015-01-01

    Tremendous research on teacher leadership over the last decade has revealed both the prevalence of and the imperatives for a model teaching force that can actively participate in school improvement (Harrison & Killion, 2007; Katzenmeyer & Moller, 2001; Leithwood & Riehl, 2003). The highly participative teacher leader paradigm is so…

  20. The Immune Pathogenesis of Immune Reconstitution Inflammatory Syndrome Associated with Highly Active Antiretroviral Therapy in AIDS

    Science.gov (United States)

    Zhou, Huaying; He, Yan; Chen, Zi; He, Bo; He, Mei

    2014-01-01

    Abstract The present study investigated the immunological pathogenesis of immune reconstitution inflammatory syndrome (IRIS) in acquired immunodeficiency syndrome (AIDS) patients undergoing highly active antiretroviral therapy (HAART). A total of 238 patients with AIDS who received initial HAART were included in this prospective cohort study. Blood samples were collected immediately, at baseline, at week 12, and at week 24 after initial HAART and at the onset of IRIS. Lymphocyte subsets, Th1 and Th2 cytokines, and interleukin (IL)-7 levels were measured by flow cytometry or ELISA. Among the 238 patients with AIDS who received HAART, 47 patients developed IRIS. The percentages of CD4+ and CD8+ naive, memory, and activated cells exhibited no significant differences between AIDS patients with and without IRIS 24 weeks after initial HAART. The percentage of CD4+CD25+Foxp3+ regulatory T cells was lower in IRIS patients than in non-IRIS patients before HAART, 12 weeks after HAART, 24 weeks after HAART, and at the onset of IRIS. IL-2 and interferon (IFN)-γ levels were significantly higher at week 4 and at the onset of IRIS in IRIS patients than in non-IRIS patients. In contrast, IL-4 and IL-10 levels were significantly lower at week 4 and at the onset of IRIS in IRIS patients than in non-IRIS patients. Plasma IL-7 decreased gradually with the progression of HAART. The level of IL-7 was higher in IRIS patients than in non-IRIS patients at all follow-up time points. An imbalance of Th1/Th2 cytokines, a consistently low CD+CD25+Fox3+ percentage, and a high IL-7 level may be crucial in the pathogenesis of IRIS in AIDS patients who had received HAART. PMID:25131160

  1. Biochemical reconstitution and phylogenetic comparison of human SET1 family core complexes involved in histone methylation.

    Science.gov (United States)

    Shinsky, Stephen A; Monteith, Kelsey E; Viggiano, Susan; Cosgrove, Michael S

    2015-03-06

    Mixed lineage leukemia protein-1 (MLL1) is a member of the SET1 family of histone H3 lysine 4 (H3K4) methyltransferases that are required for metazoan development. MLL1 is the best characterized human SET1 family member, which includes MLL1-4 and SETd1A/B. MLL1 assembles with WDR5, RBBP5, ASH2L, DPY-30 (WRAD) to form the MLL1 core complex, which is required for H3K4 dimethylation and transcriptional activation. Because all SET1 family proteins interact with WRAD in vivo, it is hypothesized they are regulated by similar mechanisms. However, recent evidence suggests differences among family members that may reflect unique regulatory inputs in the cell. Missing is an understanding of the intrinsic enzymatic activities of different SET1 family complexes under standard conditions. In this investigation, we reconstituted each human SET1 family core complex and compared subunit assembly and enzymatic activities. We found that in the absence of WRAD, all but one SET domain catalyzes at least weak H3K4 monomethylation. In the presence of WRAD, all SET1 family members showed stimulated monomethyltransferase activity but differed in their di- and trimethylation activities. We found that these differences are correlated with evolutionary lineage, suggesting these enzyme complexes have evolved to accomplish unique tasks within metazoan genomes. To understand the structural basis for these differences, we employed a "phylogenetic scanning mutagenesis" assay and identified a cluster of amino acid substitutions that confer a WRAD-dependent gain-of-function dimethylation activity on complexes assembled with the MLL3 or Drosophila trithorax proteins. These results form the basis for understanding how WRAD differentially regulates SET1 family complexes in vivo. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  2. Identification of mitochondrial carriers in Saccharomyces cerevisiae by transport assay of reconstituted recombinant proteins.

    Science.gov (United States)

    Palmieri, Ferdinando; Agrimi, Gennaro; Blanco, Emanuela; Castegna, Alessandra; Di Noia, Maria A; Iacobazzi, Vito; Lasorsa, Francesco M; Marobbio, Carlo M T; Palmieri, Luigi; Scarcia, Pasquale; Todisco, Simona; Vozza, Angelo; Walker, John

    2006-01-01

    The inner membranes of mitochondria contain a family of carrier proteins that are responsible for the transport in and out of the mitochondrial matrix of substrates, products, co-factors and biosynthetic precursors that are essential for the function and activities of the organelle. This family of proteins is characterized by containing three tandem homologous sequence repeats of approximately 100 amino acids, each folded into two transmembrane alpha-helices linked by an extensive polar loop. Each repeat contains a characteristic conserved sequence. These features have been used to determine the extent of the family in genome sequences. The genome of Saccharomyces cerevisiae contains 34 members of the family. The identity of five of them was known before the determination of the genome sequence, but the functions of the remaining family members were not. This review describes how the functions of 15 of these previously unknown transport proteins have been determined by a strategy that consists of expressing the genes in Escherichia coli or Saccharomyces cerevisiae, reconstituting the gene products into liposomes and establishing their functions by transport assay. Genetic and biochemical evidence as well as phylogenetic considerations have guided the choice of substrates that were tested in the transport assays. The physiological roles of these carriers have been verified by genetic experiments. Various pieces of evidence point to the functions of six additional members of the family, but these proposals await confirmation by transport assay. The sequences of many of the newly identified yeast carriers have been used to characterize orthologs in other species, and in man five diseases are presently known to be caused by defects in specific mitochondrial carrier genes. The roles of eight yeast mitochondrial carriers remain to be established.

  3. Green proteorhodopsin reconstituted into nanoscale phospholipid bilayers (nanodiscs) as photoactive monomers.

    Science.gov (United States)

    Ranaghan, Matthew J; Schwall, Christine T; Alder, Nathan N; Birge, Robert R

    2011-11-16

    Over 4000 putative proteorhodopsins (PRs) have been identified throughout the oceans and seas of the Earth. The first of these eubacterial rhodopsins was discovered in 2000 and has expanded the family of microbial proton pumps to all three domains of life. With photophysical properties similar to those of bacteriorhodopsin, an archaeal proton pump, PRs are also generating interest for their potential use in various photonic applications. We perform here the first reconstitution of the minimal photoactive PR structure into nanoscale phospholipid bilayers (nanodiscs) to better understand how protein-protein and protein-lipid interactions influence the photophysical properties of PR. Spectral (steady-state and time-resolved UV-visible spectroscopy) and physical (size-exclusion chromatography and electron microscopy) characterization of these complexes confirms the preparation of a photoactive PR monomer within nanodiscs. Specifically, when embedded within a nanodisc, monomeric PR exhibits a titratable pK(a) (6.5-7.1) and photocycle lifetime (∼100-200 ms) that are comparable to the detergent-solubilized protein. These ndPRs also produce a photoactive blue-shifted absorbance, centered at 377 or 416 nm, that indicates that protein-protein interactions from a PR oligomer are required for a fast photocycle. Moreover, we demonstrate how these model membrane systems allow modulation of the PR photocycle by variation of the discoidal diameter (i.e., 10 or 12 nm), bilayer thickness (i.e., 23 or 26.5 Å), and degree of saturation of the lipid acyl chain. Nanodiscs also offer a highly stable environment of relevance to potential device applications.

  4. Regulation of antiapoptotic MCL-1 function by gossypol: mechanistic insights from in vitro reconstituted systems.

    Science.gov (United States)

    Etxebarria, Aitor; Landeta, Olatz; Antonsson, Bruno; Basañez, Gorka

    2008-12-01

    Small-molecule drugs that induce apoptosis in tumor cells by activation of the BCL-2-regulated mitochondrial outer membrane permeabilization (MOMP) pathway hold promise for rational anticancer therapies. Accumulating evidence indicates that the natural product gossypol and its derivatives can kill tumor cells by targeting antiapoptotic BCL-2 family members in such a manner as to trigger MOMP. However, due to the inherent complexity of the cellular apoptotic network, the precise mechanisms by which interactions between gossypol and individual BCL-2 family members lead to MOMP remain poorly understood. Here, we used simplified systems bearing physiological relevance to examine the impact of gossypol on the function of MCL-1, a key determinant for survival of various human malignancies that has become a highly attractive target for anticancer drug design. First, using a reconstituted liposomal system that recapitulates basic aspects of the BCL-2-regulated MOMP pathway, we demonstrate that MCL-1 inhibits BAX permeabilizing function via a "dual-interaction" mechanism, while submicromolar concentrations of gossypol reverse MCL-1-mediated inhibition of functional BAX activation. Solution-based studies showed that gossypol competes with BAX/BID BH3 ligands for binding to MCL-1 hydrophobic groove, thereby providing with a mechanistic explanation for how gossypol restores BAX permeabilizing function in the presence of MCL-1. By contrast, no evidence was found indicating that gossypol transforms MCL-1 into a BAX-like pore-forming molecule. Altogether, our findings validate MCL-1 as a direct target of gossypol, and highlight that making this antiapoptotic protein unable to inhibit BAX-driven MOMP may represent one important mechanism by which gossypol exerts its cytotoxic effect in selected cancer cells.

  5. Selective expansion of viral variants following experimental transmission of a reconstituted feline immunodeficiency virus quasispecies.

    Directory of Open Access Journals (Sweden)

    Brian J Willett

    Full Text Available Following long-term infection with virus derived from the pathogenic GL8 molecular clone of feline immunodeficiency virus (FIV, a range of viral variants emerged with distinct modes of interaction with the viral receptors CD134 and CXCR4, and sensitivities to neutralizing antibodies. In order to assess whether this viral diversity would be maintained following subsequent transmission, a synthetic quasispecies was reconstituted comprising molecular clones bearing envs from six viral variants and its replicative capacity compared in vivo with a clonal preparation of the parent virus. Infection with either clonal (Group 1 or diverse (Group 2 challenge viruses, resulted in a reduction in CD4+ lymphocytes and an increase in CD8+ lymphocytes. Proviral loads were similar in both study groups, peaking by 10 weeks post-infection, a higher plateau (set-point being achieved and maintained in study Group 1. Marked differences in the ability of individual viral variants to replicate were noted in Group 2; those most similar to GL8 achieved higher viral loads while variants such as the chimaeras bearing the B14 and B28 Envs grew less well. The defective replication of these variants was not due to suppression by the humoral immune response as virus neutralising antibodies were not elicited within the study period. Similarly, although potent cellular immune responses were detected against determinants in Env, no qualitative differences were revealed between animals infected with either the clonal or the diverse inocula. However, in vitro studies indicated that the reduced replicative capacity of variants B14 and B28 in vivo was associated with altered interactions between the viruses and the viral receptor and co-receptor. The data suggest that viral variants with GL8-like characteristics have an early, replicative advantage and should provide the focus for future vaccine development.

  6. Device and method for unfastening and lifting a top nozzle subassembly from a reconstitutable fuel assembly

    International Nuclear Information System (INIS)

    Wilson, J.F.

    1987-01-01

    This patent describes a reconstitutable fuel assembly including at least one guide thimble having an upper end portion and a top nozzle subassembly having a lower adapter plate with at least one opening, and an upper hold-down plate with at least one passageway positioned above and aligned with the lower adapter plate opening. At least one hold-down spring is disposed and extends between the upper and lower plates and at least one elongated tubular hollow sleeve is disposed and extends between the upper and lower plates. The upper end portion of the guide thimble extends upwardly through the opening in the lower adapter plate and has a threaded terminal end disposed above the adapter plate. The threaded terminal end of the guide thimble and an upper end extend upwardly through the passageway of the upper hold-down plate. A device is described for unfastening and lifting the top nozzle subassembly from the guide thimble of the fuel assembly, comprising: (a) at least one hollow gripper tube, the tube having an open lower end; (b) means mounting the gripper tube for vertical alignment with and insertion of its lower end portion into the elongated sleeve of the top nozzle subassembly to a position therein located above and adjacent to the threaded lower end of the sleeve; (c) force-generating means disposed within the gripper tube for rotatable movement and concurrent axial movement upwardly and downwardly within the tube and also disposed at the open lower end of the gripper tube for extension into and from the gripper tube open lower end upon axial movement upwardly and downwardly within the gripper tube

  7. Fixture and method for rectifying damaged guide thimble insert sleeves in a reconstitutable fuel assembly

    International Nuclear Information System (INIS)

    Shallenberger, J.M.; Ferlan, S.J.

    1987-01-01

    A guide thimble damage-rectifying method is described for use on a reconstitutable fuel assembly being held in a work station with its top nozzle removed to expose a plurality of guide thimbles having one of several different types of damage. The method consists of: (a) providing a base having a plurality of tool positioning openings defined therein in a pattern matched with that of the guide thimbles of the fuel assembly; (b) mounting the base on the work station with its tool positioning openings in alignment with the guide thimbles of the fuel assembly and such that the base is movable toward the guide thimbles; (c) providing a plurality of different tools each operable to rectify one of the different types of guide thimble damage; (d) mounting selected ones of the different tools in respective ones of the openings of the base in alignment with ones of the thimbles having the respective types of guide thimble damage capable of being rectified by the selected tools such that upon movement of the base toward the guide thimbles the respective types of guide thimble damage will be rectified by the selected tools; (e) providing a group of positioning elements; (f) mounting the positioning elements in selected ones of the base openings corresponding to undamaged ones of the guide thimbles such that upon movement of the base toward the guide thimbles the positioning elements become mounted on upper end portions of the corresponding undamaged ones of the guide thimbles for precisely locating the fixture relative to the guide thimble upper end portions for accurate performance of the repairable damage rectifying operation by the tools as the base is moved toward the guide thimbles; and (g) moving the base toward the guide thimbles so as to mount the positioning elements on the corresponding ones of the undamaged guide thimbles and effect rectification of the damaged guide thimbles by the selected tools

  8. Inductive differentiation of two neural lineages reconstituted in a microculture system from Xenopus early gastrula cells.

    Science.gov (United States)

    Mitani, S; Okamoto, H

    1991-05-01

    Neural induction of ectoderm cells has been reconstituted and examined in a microculture system derived from dissociated early gastrula cells of Xenopus laevis. We have used monoclonal antibodies as specific markers to monitor cellular differentiation from three distinct ectoderm lineages in culture (N1 for CNS neurons from neural tube, Me1 for melanophores from neural crest and E3 for skin epidermal cells from epidermal lineages). CNS neurons and melanophores differentiate when deep layer cells of the ventral ectoderm (VE, prospective epidermis region; 150 cells/culture) and an appropriate region of the marginal zone (MZ, prospective mesoderm region; 5-150 cells/culture) are co-cultured, but not in cultures of either cell type on their own; VE cells cultured alone yield epidermal cells as we have previously reported. The extent of inductive neural differentiation in the co-culture system strongly depends on the origin and number of MZ cells initially added to culture wells. The potency to induce CNS neurons is highest for dorsal MZ cells and sharply decreases as more ventrally located cells are used. The same dorsoventral distribution of potency is seen in the ability of MZ cells to inhibit epidermal differentiation. In contrast, the ability of MZ cells to induce melanophores shows the reverse polarity, ventral to dorsal. These data indicate that separate developmental mechanisms are used for the induction of neural tube and neural crest lineages. Co-differentiation of CNS neurons or melanophores with epidermal cells can be obtained in a single well of co-cultures of VE cells (150) and a wide range of numbers of MZ cells (5 to 100). Further, reproducible differentiation of both neural lineages requires intimate association between cells from the two gastrula regions; virtually no differentiation is obtained when cells from the VE and MZ are separated in a culture well. These results indicate that the inducing signals from MZ cells for both neural tube and neural

  9. Validation of a published case definition for tuberculosis-associated immune reconstitution inflammatory syndrome.

    Science.gov (United States)

    Haddow, Lewis J; Moosa, Mahomed-Yunus S; Easterbrook, Philippa J

    2010-01-02

    To evaluate the International Network for the Study of HIV-associated IRIS (INSHI) case definitions for tuberculosis (TB)-associated immune reconstitution inflammatory syndrome (IRIS) in a South African cohort. Prospective cohort of 498 adult HIV-infected patients initiating antiretroviral therapy. Patients were followed up for 24 weeks and all clinical events were recorded. Events with TB-IRIS as possible cause were assessed by consensus expert opinion and INSHI case definition. Positive, negative, and chance-corrected agreement (kappa) were calculated, and reasons for disagreement were assessed. One hundred and two (20%) patients were receiving TB therapy at antiretroviral therapy initiation. Three hundred and thirty-three events were evaluated (74 potential paradoxical IRIS, 259 potential unmasking IRIS). Based on expert opinion, there were 18 cases of paradoxical IRIS associated with TB and/or other opportunistic disease. The INSHI criteria for TB-IRIS agreed in 13 paradoxical cases, giving positive agreement of 72.2%, negative agreement in 52/56 non-TB-IRIS events (92.9%), and kappa of 0.66. There were 19 unmasking TB-IRIS cases based on expert opinion, of which 12 were considered IRIS using the INSHI definition (positive agreement 63.2%). There was agreement in all 240 non-TB-IRIS events (negative agreement 100%) and kappa was 0.76. There was good agreement between the INSHI case definition for both paradoxical and unmasking TB-IRIS and consensus expert opinion. These results support the use of this definition in clinical and research practice, with minor caveats in its application.

  10. In vitro reconstitution and characterization of the yeast mitochondrial degradosome complex unravels tight functional interdependence.

    Science.gov (United States)

    Malecki, Michal; Jedrzejczak, Robert; Stepien, Piotr P; Golik, Pawel

    2007-09-07

    The mitochondrial degradosome (mtEXO), the main RNA-degrading complex of yeast mitochondria, is composed of two subunits: an exoribonuclease encoded by the DSS1 gene and an RNA helicase encoded by the SUV3 gene. We expressed both subunits of the yeast mitochondrial degradosome in Escherichia coli, reconstituted the complex in vitro and analyzed the RNase, ATPase and helicase activities of the two subunits separately and in complex. The results reveal a very strong functional interdependence. For every enzymatic activity, we observed significant changes when the relevant protein was present in the complex, compared to the activity measured for the protein alone. The ATPase activity of Suv3p is stimulated by RNA and its background activity in the absence of RNA is reduced greatly when the protein is in the complex with Dss1p. The Suv3 protein alone does not display RNA-unwinding activity and the 3' to 5' directional helicase activity requiring a free 3' single-stranded substrate becomes apparent only when Suv3p is in complex with Dss1p. The Dss1 protein alone does have some basal exoribonuclease activity, which is not ATP-dependent, but in the presence of Suv3p the activity of the entire complex is enhanced greatly and is entirely ATP-dependent, with no residual activity observed in the absence of ATP. Such absolute ATP-dependence is unique among known exoribonuclease complexes. On the basis of these results, we propose a model in which the Suv3p RNA helicase acts as a molecular motor feeding the substrate to the catalytic centre of the RNase subunit.

  11. Reconstitution of dewatered food processing residuals with manure to increase energy production from anaerobic digestion

    International Nuclear Information System (INIS)

    Wall, David M.; Wu-Haan, Wei; Safferman, Steven I.

    2012-01-01

    Solid residuals generated from dewatering food processing wastewater contain organic carbon that can potentially be reclaimed for energy through anaerobic digestion. This results in the diversion of waste from a landfill and uses it for a beneficial purpose. Dewatering the waste concentrates the carbon, reducing transportation costs to a farm digester where it can be blended with manure to increase biogas yield. Polymers are often used in the dewatering of the food waste but little is known regarding their impact on biogas production. Four 2 dm 3 working volume, semi-continuous reactors, were used at a mesophilic temperature and a solids retention time (SRT) of 15 days. Reactors were fed daily with a blended feedstock containing a food processing sludge waste (FPSW)/manure ratio of 2.2:1 (by weight) as this produced the optimized carbon to nitrogen ratio. Results demonstrated that reconstitution of dewatered FPSW with dairy manure produced approximately 2 times more methane than animal manure alone for the same volume. However, only approximately 30% of volatile solids (VS) were consumed indicating energy potential still remained. Further, the efficiency of the conversion of VS to methane for the blended FPSW/manure was substantially less than for manure only. However, the overall result is an increase in energy production for a given tank volume, which can decrease life cycle costs. Because all FPSW is unique and the determination of dewatering additives is customized based on laboratory testing and field adjustment, generalizations are difficult and specific testing is required. -- Highlights: ► Energy production in anaerobic digestion can increase by co-blending food waste. ► Energy for transporting food waste to blend with manure is less when dewatered. ► Dewatered food waste in manure produced twice as much methane than manure. ► Efficiency of carbon to methane was low because of ammonium bicarbonate production. ► Carbon destruction was 30%, more

  12. Immune reconstitution inflammatory syndrome in HIV-infected patients with Pneumocystis jirovecii pneumonia.

    Science.gov (United States)

    Roade Tato, Luisa; Burgos Cibrian, Joaquín; Curran Fábregas, Adrià; Navarro Mercadé, Jordi; Willekens, Rein; Martín Gómez, María Teresa; Ribera Pascuet, Esteban; Falcó Ferrer, Vicenç

    2017-11-26

    The incidence of immune reconstitution inflammatory syndrome (IRIS) in HIV-infected patients after an episode of Pneumocystis jirovecii pneumonia (PJP) seems to be lower than with other opportunistic infections. We conducted an observational study in order to determine the incidence, clinical characteristics and outcome of patients diagnosed with PJP-related IRIS. We conducted an observational study of HIV patients diagnosed with PJP-related IRIS from January 2000 to November 2015. We analyzed epidemiological and clinical characteristics as well as laboratory findings. We also carried out a systematic review of published cases. Six cases of IRIS out of 123 (4.9%) HIV-infected patients with PJP who started ART were diagnosed. All six cases were men with a median age of 34 (IQR: 8) years. The six patients developed paradoxical IRIS. Subjects younger than 40 years old (p=0.084) and with an HIV-RNA viral load >100000 copies/ml (p=0.081) at diagnosis showed a tendency to develop IRIS. Thirty-seven published cases of PJP-related IRIS were identified. Although 51% of cases involved respiratory failure, no deaths were reported. PJP-related IRIS is rare condition compared to other opportunistic infections. It can lead to a severe respiratory failure in a significant proportion of cases, although no deaths have been reported. Copyright © 2017 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  13. Home in the heat: Dramatic seasonal variation in home range of desert golden eagles informs management for renewable energy development

    Science.gov (United States)

    Melissa Braham; Tricia Miller; Adam E. Duerr; Michael Lanzone; Amy Fesnock; Larry LaPre; Daniel Driscoll; Todd. Katzner

    2015-01-01

    Renewable energy is expanding quickly with sometimes dramatic impacts to species and ecosystems. To understand the degree to which sensitive species may be impacted by renewable energy projects, it is informative to know how much space individuals use and how that space may overlap with planned development. We used global positioning system-global system for mobile...

  14. The Development of Evaluation Model for Internal Quality Assurance System of Dramatic Arts College of Bunditpattanasilpa Institute

    Science.gov (United States)

    Sinthukhot, Kittisak; Srihamongkol, Yannapat; Luanganggoon, Nuchwana; Suwannoi, Paisan

    2013-01-01

    The research purpose was to develop an evaluation model for the internal quality assurance system of the dramatic arts College of Bunditpattanasilpa Institute. The Research and Development method was used as research methodology which was divided into three phases; "developing the model and its guideline", "trying out the actual…

  15. Learning Form and Function by Dance-Dramatizing Cultural Legends to Drum Rhythms Wearing Student-Made Animal Masks

    Science.gov (United States)

    Gray, Phyllis; Rule, Audrey C.; Kirkland Holmes, Gloria; Logan, Stephanie R.; Alert, Andrea L.; Mason, Cynthia A.

    2016-01-01

    This study examined the self-efficacy in science, art, dance, and music; attitudes concerning contributions of people of various ethnic/cultural groups; and science learning of students involved in an after-school arts-integrated science enrichment project. Students dramatized three traditional animal legends from African, Native American, and…

  16. TAR (Theatre as Representation) as a Provocative Teaching Tool in School Administration: A Dramatized Inclusive Classroom Scenario

    Science.gov (United States)

    Meyer, Matthew J.; Young, David C.

    2013-01-01

    The following dramatized classroom scenario depicts a teacher struggling with the nature of an inclusive learning environment, with instructional leadership and supervision of instruction as the theoretical and practical backdrop. The purpose of this article is to demonstrate how the use of a TAR (theatre as representation) case study can be used…

  17. 30 years of data reveal dramatic increase in abundance of brown trout following the removal of a small hydrodam

    DEFF Research Database (Denmark)

    Birnie-Gauvin, Kim; Larsen, Martin Hage; Nielsen, Jan

    2017-01-01

    and spawning of fish. In the present study, we present thirty years of data from electrofishing surveys obtained at two sites, both prior to and following the removal of a small-scale hydropower dam in Central Jutland, Denmark. We demonstrate that the dam removal has led to a dramatic increase in trout density...

  18. Emergence of dramatic literary form in the classical period and its effect on the English Renaissance drama

    Directory of Open Access Journals (Sweden)

    Andrejević Ana M.

    2010-01-01

    Full Text Available In the literary-historical and theoretical sense, the ancient drama established the initial genre of drama as the basis for its future evolution. Despite the poetic differences in understanding the certain elements of drama, ancient authors established a basis for continuity of development of European drama in future centuries, literary periods and stylistic formations. Diachronic perspective of the drama development in English literature will also play a certain role in the centuries that follow. However, the existence of a rich national dramatic tradition in England had, after all, greater impact on dramatic literature, which will allow classical models only to steer the development of drama in the right direction, not allowing the authors to be captured in rules and uniformity. University wits, with their adaptation of ancient drama conventions and their combination with traditional medieval elements, created the model for a comedy and a tragedy which needed only a natural skill of a genius to use them in proportion and to raise them to a higher level of dramatic art. Then, fortunately, William Shakespeare, who will achieve that aesthetic-axiological dramatic scale, appears. Like many great writers, Shakespeare was not a founder of a new tradition, but the culmination of an existing one, like Veselin Kostić put it. The appearance of the classical interpretation of drama and great interest in Seneca, Terence and Plautus quickly found its way from the academic circles to the national theaters and Shakespeare did not remain immune to these influences.

  19. Social insect genomes exhibit dramatic evolution in gene composition and regulation while preserving regulatory features linked to sociality

    DEFF Research Database (Denmark)

    Simola, Daniel F.; Wissler, Lothar; Donahue, Greg

    2013-01-01

    Genomes of eusocial insects code for dramatic examples of phenotypic plasticity and social organization. We compared the genomes of seven ants, the honeybee, and various solitary insects to examine whether eusocial lineages share distinct features of genomic organization. Each ant lineage contain...

  20. Activation and binding of opsonic fragments of C3 on encapsulated Cryptococcus neoformans by using an alternative complement pathway reconstituted from six isolated proteins.

    Science.gov (United States)

    Kozel, T R; Wilson, M A; Pfrommer, G S; Schlageter, A M

    1989-07-01

    Encapsulated Cryptococcus neoformans yeast cells are potent activators of the complement system. We examined the interaction of the yeast cells with an alternative complement pathway reconstituted from isolated factor D, factor B, factor H, factor I, C3, and properdin. Incubation of encapsulated cryptococci with the reconstituted pathway led to activation and binding of C3 fragments to the yeast cells that was quantitatively and qualitatively identical to that observed with normal human serum. Incubation with either normal serum or a mixture of isolated proteins led to binding of 4 x 10(7) to 5 x 10(7) C3 molecules to the yeast cells. The kinetics for activation and binding of C3 were identical, with maximum binding observed after a 20-min incubation. Immunoglobulin G was not needed for optimal activation kinetics. C3 fragments eluted from the yeast cells by treatment with hydroxylamine and subsequent analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated the presence primarily of iC3b on yeast cells incubated with either normal serum or the reconstituted pathway. Ultrastructural examination of the opsonized yeast cells showed that the cryptococcal capsule was the site for binding of C3 activated from normal serum or the reconstituted pathway, with a dense accumulation of C3 at the periphery of the capsule. Thus, incubation of encapsulated cryptococci in the reconstituted pathway led to deposition of opsonic complement fragments at a site that was appropriate for interaction with phagocyte receptors. Cryptococci opsonized with the reconstituted pathway showed a markedly enhanced interaction with cultured human monocytes compared with unopsonized yeast cells, indicating that the alternative pathway alone is opsonic for yeast cells. However, the results indicate that additional serum factors are needed for optimal opsonization of yeast cells because a 35% reduction in the number of cryptococci bound to macrophages was observed with

  1. Spectroscopic investigation of the aggregation state of amphotericin B during loading, freeze-drying, and reconstitution of polymeric micelles.

    Science.gov (United States)

    Adams, Monica; Kwon, Glen S

    2004-11-22

    To investigate the relative aggregation state of amphotericin B (AmB) during loading and reconstitution of polymeric micelles. Hexanoate and stearate derivatives of PEO-b-p (L-Asp) were prepared. The polymers and AmB were dissolved in methanol (MeOH). Milli-Q water was then added slowly, and the MeOH was removed via rotary evaporation. The solutions were freeze-dried in the presence of trehalose. During micelle preparation, the aggregation state of AmB was assessed using absorption spectroscopy. Upon reconstitution, the samples were analyzed using vapor-pressure osmometry, size-exclusion chromatography (SEC), and absorption spectroscopy. The absorption spectrum of AmB in the presence of the block copolymers was compared to that of AmB alone under the same conditions. AmB was loaded into micelles prepared from acyl derivatives of PEO-b-p (L-Asp). Absorption spectroscopy indicated that the aggregation state was preserved during the loading process. AmB exists in a self-aggregated state in polymeric micelles containing hexanoate ester cores and in a relatively monomeric state in polymeric micelles containing stearate ester cores. Vapor-pressure osmometry confirmed the isotonicity of the formulations, while SEC indicated that the micelles were approximately 10(6) g/mol. Depending on the polymer structure and assembly conditions, it is possible to encapsulate AmB in a relatively nonaggregated or aggregated state in micelles prepared from acyl derivatives of PEO-b-p (L-Asp). In polymeric micelles containing stearate side chains, AmB was loaded in a nearly monomeric state, possibly due to interaction with the stearate side chains. The final aggregation state of the drug is preserved during lyophilization and reconstitution of polymeric micelles prepared by a novel solvent evaporation procedure.

  2. Long-term leukocyte reconstitution in NSG mice transplanted with human cord blood hematopoietic stem and progenitor cells.

    Science.gov (United States)

    Audigé, Annette; Rochat, Mary-Aude; Li, Duo; Ivic, Sandra; Fahrny, Audrey; Muller, Christina K S; Gers-Huber, Gustavo; Myburgh, Renier; Bredl, Simon; Schlaepfer, Erika; Scherrer, Alexandra U; Kuster, Stefan P; Speck, Roberto F

    2017-05-30

    Humanized mice (hu mice) are based on the transplantation of hematopoietic stem and progenitor cells into immunodeficient mice and have become important pre-clinical models for biomedical research. However, data about their hematopoiesis over time are scarce. We therefore characterized leukocyte reconstitution in NSG mice, which were sublethally irradiated and transplanted with human cord blood-derived CD34+ cells at newborn age, longitudinally in peripheral blood and, for more detailed analyses, cross-sectionally in peripheral blood, spleen and bone marrow at different time points. Human cell chimerism and absolute human cell count decreased between week 16 and 24 in the peripheral blood of hu mice, but were stable thereafter as assessed up to 32 weeks. Human cell chimerism in spleen and bone marrow was maintained over time. Notably, human cell chimerism in peripheral blood and spleen as well as bone marrow positively correlated with each other. Percentage of B cells decreased between week 16 and 24, whereas percentage of T cells increased; subsequently, they levelled off with T cells clearly predominating at week 32. Natural killer cells, monocytes and plasmacytoid dendritic cells (DCs) as well as CD1c + and CD141+ myeloid DCs were all present in hu mice. Proliferative responses of splenic T cells to stimulation were preserved over time. Importantly, the percentage of more primitive hematopoietic stem cells (HSCs) in bone marrow was maintained over time. Overall, leukocyte reconstitution was maintained up to 32 weeks post-transplantation in our hu NSG model, possibly explained by the maintenance of HSCs in the bone marrow. Notably, we observed great variation in multi-lineage hematopoietic reconstitution in hu mice that needs to be taken into account for the experimental design with hu mice.

  3. ZVI Addition in Continuous Anaerobic Digestion Systems Dramatically Decreases P Recovery Potential: Dynamic Modelling

    DEFF Research Database (Denmark)

    Puyol, D.; Flores Alsina, Xavier; Segura, Y.

    2017-01-01

    The objective of this study is to show the preliminary results of a (dynamic) mathematical model describing the effects of zero valent iron (ZVI) addition during the anaerobic digestion of waste activated sludge from wastewater treatment systems. A modified version of the Anaerobic Digestion Model...... No. 1 (ADM1) upgraded with an improved physico-chemical description, ZVI corrosion, propionate uptake enhancement and multiple mineral precipitation is used as a modelling platform. The proposed approach is tested against two case studies which correspond to two lab scale anaerobic digesters (AD2, AD....... Simulations demonstrate that the model is capable to satisfactorily reproduce the dynamics of hydrolysis, acetogenesis, acidogenesis, nutrient release, pH and methanogenesis in the control anaerobic digester (AD1). This study also evidences the enhancement of methane production by the influence of ZVI...

  4. Continuous quality improvement

    International Nuclear Information System (INIS)

    Bourne, P.B.

    1985-01-01

    This paper describes the various statistical tools used at the Hanford Engineering Development Laboratory to achieve continuous quality improvement in the development of Breeder Reactor Technology and in reactor operations. The role of the quality assurance professionals in this process, including quantifiable measurements using actual examples, is provided. The commitment to quality improvement through top management involvement is dramatically illustrated

  5. Induced polyploidy dramatically increases the size and alters the shape of fruit in Actinidia chinensis

    Science.gov (United States)

    Wu, Jin-Hu; Ferguson, A. Ross; Murray, Brian G.; Jia, Yilin; Datson, Paul M.; Zhang, Jingli

    2012-01-01

    Background and Aims Some otherwise promising selections of Actinidia chinensis (kiwifruit) have fruit that are too small for successful commercialization. We have therefore made the first detailed study in diploid kiwifruit of the effects of chromosome doubling induced by colchicine on fruit size, shape and crop loading. Methods Flow cytometric analysis of young leaves and chromosome analysis of flower buds and root tips was used to confirm the stability of induced autotetraploids. Fruit weight, size and crop load were measured in the third year after planting in the field and for three consecutive years. DNA fingerprinting was used to confirm the origin of the material. Key Results There was a very significant increase in fruit size in induced autotetraploids of different genotypes of A. chinensis. With the commercially important diploid cultivar ‘Hort16A’, most regenerants, Type A plants, had fruit which were much the same shape as fruit of the diploid but, at the same fruit load, were much larger and heavier. Some regenerants, Type B plants, produced fruit similar to ‘fasciated’ fruit. Fruit of the autotetraploids induced from three female red-fleshed A. chinensis selections were also 50–60 % larger than fruit of their diploid progenitors. The main increase in fruit dimensions was in their diameters. These improved fruit characteristics were stable over several seasons. Conclusions Chromosome doubling has been shown to increase significantly fruit size in autotetraploid A. chinensis, highlighting the considerable potential of this technique to produce new cultivars with fruit of adequate size. Other variants with differently shaped fruit were also produced but the genetic basis of this variation remains to be elucidated. Autoploids of other Actinidia species with commercial potential may also show improved fruit characteristics, opening up many new possibilities for commercial development. PMID:21980192

  6. Paradoxical immune reconstitution inflammatory syndrome associated with previous Cryptococcus neoformans infection in an HIV-positive patient requiring neurosurgical intervention.

    Science.gov (United States)

    Biagetti, Carlo; Nicola, Monica; Borderi, Marco; Pavoni, Michele; Tampellini, Livia; Verucchi, Gabriella; Chiodo, Francesco

    2009-04-01

    Immune reconstitution inflammatory syndrome (IRIS) in HIV-1-infected patients is associated with an exaggerated inflammatory response against an opportunistic infection during highly active antiretroviral therapy. The only review on IRIS associated with Criptococcus neoformans reported 21 episodes including lymphadenitis, necrotizing pneumonitis, breast and cutaneous abscess, and cryptococcomas. To our knowledge this is the first report of IRIS associated with previous meningeal criptococcal infection which required neurosurgical intervention with placement of a ventriculo-peritoneal shunt to drain a CSF cyst formed by exclusion of the temporal horn of the right lateral ventricle. We demonstrate that this procedure is possible without complications such as cryptococcal dissemination into the peritoneum.

  7. Cryptococcal breast abscess in an HIV-positive patient: arguments for reviewing the definition of immune reconstitution inflammatory syndrome.

    Science.gov (United States)

    Haddow, Lewis J; Sahid, Faieza; Moosa, Mahomed-Yunus S

    2008-07-01

    Atypical manifestations of Cryptococcus neoformans disease have been reported in patients with HIV-1 infection as part of the spectrum of the immune reconstitution inflammatory syndrome (IRIS). We describe a cryptococcal breast abscess in a patient presenting after 11 months of highly active antiretroviral therapy (HAART). The arguments for and against the case being a novel manifestation of IRIS are discussed. The potential hazards of using CD4 count as a surrogate marker of IRIS and the danger of misdiagnosing IRIS as failure of HAART are highlighted.

  8. Children's experiences of the repositioning of their psychological birth order in a reconstituted family / Lizelle van Jaarsveld.

    OpenAIRE

    Van Jaarsveld, Lizelle

    2012-01-01

    The aim of this study is to explore and describe children’s experiences of the repositioning of their psychological birth order in a reconstituted family. The aim of this study is also to contribute to a better understanding of this phenomenon to aid the parents of these children as well as professionals working with such families. The systems theory was used as the meta-theory of this study. Gestalt field theory formed the connection between the systems theory and Adlerian theory, to port...

  9. Electrochemical studies of a reconstituted photosynthetic electron-transfer chain or towards a biomimetic photoproduction of hydrogen

    International Nuclear Information System (INIS)

    Fourmond, V.

    2007-04-01

    The aim of this work is to find an efficient process to convert solar energy into hydrogen. The electrons transfers in reconstituted photosynthetic chains have been particularly studied with the aims 1)in one hand, to better understand the interactions of the different molecules of the photosynthetic chain in order to optimize the changes of the entire organisms for hydrogen production 2)in another hand, to insert the hydrogenases in a photosynthetic chain and then to photo reduce them in order to obtain kinetic data to better understand how it works. (O.M.)

  10. Combined effects of plant extracts in inhibiting the growth of Bacillus cereus in reconstituted infant rice cereal.

    Science.gov (United States)

    Jun, Hyejung; Kim, Jinsol; Bang, Jihyun; Kim, Hoikyung; Beuchat, Larry R; Ryu, Jee-Hoon

    2013-01-01

    A study was done to determine the potential use of plant extracts to inhibit the growth of Bacillus cereus in reconstituted infant rice cereal. A total of 2116 extracts were screened for inhibitory activity against B. cereus using an agar well diffusion assay. The minimal inhibitory concentrations (MIC) and minimal lethal concentrations (MLC) of 14 promising extracts in tryptic soy broth (TSB) were determined. Dryopteris erythrosora (autumn fern) root extract showed the lowest MIC (0.0156 mg/ml), followed by Siegesbeckia glabrescens (Siegesbeckia herb) leaf (0.0313 mg/ml), Morus alba (white mulberry) cortex (0.0313 mg/ml), Carex pumila (sand sedge) root (0.0625 mg/ml), and Citrus paradisi (grapefruit) seed (0.0625 mg/ml) extracts. The order of MLCs of extracts was D. erythrosora root (0.0156 mg/ml)extracts against B. cereus in TSB were determined using a checkerboard assay. A combination of D. erythrosora and C. pumila extracts showed a partial synergistic inhibition, with a fractional inhibitory concentration index (FICI) of 0.75. Single and combined inhibitory activities of selected plant extracts against B. cereus in reconstituted infant rice cereal were investigated. The MICs of S. glabrescens, M. alba, D. erythrosora, and C. pumila extracts against B. cereus were 1.0, 2.0, 2.0, and 8.0mg/ml, respectively. A combination of D. erythrosora (1.00 mg/ml) and C. pumila (1.00 mg/ml) extracts showed a partial synergistic effect (FICI 0.63) in inhibiting the growth of B. cereus. Results indicate that by combining extracts, the amounts of D. erythrosora and C. pumila extracts can be reduced by 50% and 87.5%, respectively, compared with individual extracts, and give similar inhibitory activity in reconstituted infant rice cereal. Sensory evaluation showed that supplementing reconstituted infant rice cereal with plant extracts reduces sensorial quality. These observations will be useful when developing and applying interventions using natural plant extracts to inhibit B

  11. Spectroscopic studies of the iron and manganese reconstituted tyrosyl radical in Bacillus cereus ribonucleotide reductase R2 protein.

    Directory of Open Access Journals (Sweden)

    Ane B Tomter

    Full Text Available Ribonucleotide reductase (RNR catalyzes the rate limiting step in DNA synthesis where ribonucleotides are reduced to the corresponding deoxyribonucleotides. Class Ib RNRs consist of two homodimeric subunits: R1E, which houses the active site; and R2F, which contains a metallo cofactor and a tyrosyl radical that initiates the ribonucleotide reduction reaction. We studied the R2F subunit of B. cereus reconstituted with iron or alternatively with manganese ions, then subsequently reacted with molecular oxygen to generate two tyrosyl-radicals. The two similar X-band EPR spectra did not change significantly over 4 to 50 K. From the 285 GHz EPR spectrum of the iron form, a g(1-value of 2.0090 for the tyrosyl radical was extracted. This g(1-value is similar to that observed in class Ia E. coli R2 and class Ib R2Fs with iron-oxygen cluster, suggesting the absence of hydrogen bond to the phenoxyl group. This was confirmed by resonance Raman spectroscopy, where the stretching vibration associated to the radical (C-O, ν(7a = 1500 cm(-1 was found to be insensitive to deuterium-oxide exchange. Additionally, the (18O-sensitive Fe-O-Fe symmetric stretching (483 cm(-1 of the metallo-cofactor was also insensitive to deuterium-oxide exchange indicating no hydrogen bonding to the di-iron-oxygen cluster, and thus, different from mouse R2 with a hydrogen bonded cluster. The HF-EPR spectrum of the manganese reconstituted RNR R2F gave a g(1-value of ∼2.0094. The tyrosyl radical microwave power saturation behavior of the iron-oxygen cluster form was as observed in class Ia R2, with diamagnetic di-ferric cluster ground state, while the properties of the manganese reconstituted form indicated a magnetic ground state of the manganese-cluster. The recent activity measurements (Crona et al., (2011 J Biol Chem 286: 33053-33060 indicates that both the manganese and iron reconstituted RNR R2F could be functional. The manganese form might be very important, as it has 8

  12. Comparative ecotoxicity of polystyrene nanoparticles in natural seawater and reconstituted seawater using the rotifer Brachionus plicatilis.

    Science.gov (United States)

    Manfra, L; Rotini, A; Bergami, E; Grassi, G; Faleri, C; Corsi, I

    2017-11-01

    The impact of nanoplastics using model polystyrene nanoparticles (PS NPs), anionic (PS-COOH) and cationic (PS-NH 2 ), has been investigated on the marine rotifer Brachionus plicatilis, a major component of marine zooplanktonic species. The role of different surface charges in affecting PS NP behaviour and toxicity has been considered in high ionic strength media. To this aim, the selected media were standardized reconstituted seawater (RSW) and natural sea water (NSW), the latter resembling more natural exposure scenarios. Hatched rotifer larvae were exposed for 24h and 48h to both PS NPs in the range of 0.5-50μg/ml using PS NP suspensions made in RSW and NSW. No effects on lethality upon exposure to anionic NPs were observed despite a clear gut retention was evident in all exposed rotifers. On the contrary, cationic NPs caused lethality to rotifer larvae but LC 50 values resulted lower in rotifers exposed in RSW (LC 50 =2.75±0.67µg/ml) compared to those exposed in NSW (LC 50 =6.62±0.87µg/ml). PS NPs showed similar pattern of aggregation in both high ionic strength media (RSW and NSW) but while anionic NPs resulted in large microscale aggregates (Z-average 1109 ± 128nm and 998±67nm respectively), cationic NP aggregates were still in nano-size forms (93.99 ± 11.22nm and 108.3 ± 12.79nm). Both PDI and Z-potential of PS NPs slightly differed in the two media suggesting a role of their different surface charges in affecting their behaviour and stability. Our findings confirm the role of surface charges in nanoplastic behaviour in salt water media and provide a first evidence of a different toxicity in rotifers using artificial media (RSW) compared to natural one (NSW). Such evidence poses the question on how to select the best medium in standardized ecotoxicity assays in order to properly assess their hazard to marine life in natural environmental scenarios. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Clinical case definition and manifestations of paradoxical tuberculosis-associated immune reconstitution inflammatory syndrome.

    Science.gov (United States)

    Manosuthi, Weerawat; Van Tieu, Hong; Mankatitham, Wiroj; Lueangniyomkul, Aroon; Ananworanich, Jintanat; Avihingsanon, Anchalee; Siangphoe, Umaporn; Klongugkara, Sukonsri; Likanonsakul, Sirirat; Thawornwan, Unchana; Suntisuklappon, Bussakorn; Sungkanuparph, Somnuek

    2009-11-27

    The International Network for the Study of HIV-associated IRIS (INSHI) recently published criteria for tuberculosis-associated immune reconstitution inflammatory syndrome (TB-IRIS) diagnosis. The performance of this definition and clinical manifestations of TB-IRIS were studied. Antiretroviral therapy-naive HIV/TB Thai patients receiving antituberculous therapy were enrolled during 2006-2007 and prospectively followed through 24 weeks of antiretroviral therapy. Patients were defined as having paradoxical TB-IRIS if they fulfilled the 'study definition' by French 2004 and were confirmed by an external reviewer. All were later compared by the classification according to 'INSHI-2008'. For the 126 patients, median baseline CD4 cell count was 43 cells/microl and HIV-1 RNA was 5.9 log(10) Y copies/ml. Seventy-three (58%) had extrapulmonary/disseminated TB. Twenty-two (18%) and 21 (17%) fulfilled TB-IRIS criteria according to the study definition and INSHI-2008 definition, respectively. Two (2%) were diagnosed by study definition only and one (1%) by INSHI-2008 definition only. Twenty (16%) were concordantly diagnosed by both definitions and 103 (82%) were consistently negative. Eighteen (82%) had worsening of a preexisting site, whereas four (18%) had TB-IRIS in a new location. Lymph node enlargement (73%) and fever (59%) were common in TB-IRIS. Sensitivity and specificity of INSHI-2008 was 91% (95% confidence interval, 72-98%) and 99% (95% confidence interval, 95-99.8%), respectively. Positive predictive value was 95% and negative predictive value was 98%. By multivariate analysis, factors predicting TB-IRIS were extrapulmonary TB (odds ratio, 8.63) and disseminated TB (odds ratio, 4.17). There was high concordance between the INSHI-2008 and French 2004 definition for TB-IRIS diagnosis in HIV/TB patients with relatively high rate of paradoxical TB-IRIS. This suggests that lack of HIV-1 RNA and CD4 cell count monitoring does not impede the ability to diagnose TB-IRIS.

  14. Structure-function relationships in reconstituted HDL: Focus on antioxidative activity and cholesterol efflux capacity.

    Science.gov (United States)

    Cukier, Alexandre M O; Therond, Patrice; Didichenko, Svetlana A; Guillas, Isabelle; Chapman, M John; Wright, Samuel D; Kontush, Anatol

    2017-09-01

    High-density lipoprotein (HDL) contains multiple components that endow it with biological activities. Apolipoprotein A-I (apoA-I) and surface phospholipids contribute to these activities; however, structure-function relationships in HDL particles remain incompletely characterised. Reconstituted HDLs (rHDLs) were prepared from apoA-I and soy phosphatidylcholine (PC) at molar ratios of 1:50, 1:100 and 1:150. Oxidative status of apoA-I was varied using controlled oxidation of Met112 residue. HDL-mediated inactivation of PC hydroperoxides (PCOOH) derived from mildly pre-oxidized low-density lipoprotein (LDL) was evaluated by HPLC with chemiluminescent detection in HDL+LDL mixtures and re-isolated LDL. Cellular cholesterol efflux was characterised in RAW264.7 macrophages. rHDL inactivated LDL-derived PCOOH in a dose- and time-dependent manner. The capacity of rHDL to both inactivate PCOOH and efflux cholesterol via ATP-binding cassette transporter A1 (ABCA1) increased with increasing apoA-I/PC ratio proportionally to the apoA-I content in rHDL. Controlled oxidation of apoA-I Met112 gradually decreased PCOOH-inactivating capacity of rHDL but increased ABCA1-mediated cellular cholesterol efflux. Increasing apoA-I content in rHDL enhanced its antioxidative activity towards oxidized LDL and cholesterol efflux capacity via ABCA1, whereas oxidation of apoA-I Met112 decreased the antioxidative activity but increased the cholesterol efflux. These findings provide important considerations in the design of future HDL therapeutics. Non-standard abbreviations and acronyms: AAPH, 2,2'-azobis(-amidinopropane) dihydrochloride; ABCA1, ATP-binding cassette transporter A1; apoA-I, apolipoprotein A-I; BHT, butylated hydroxytoluene; CV, cardiovascular; EDTA, ethylenediaminetetraacetic acid; HDL-C, high-density lipoprotein cholesterol; LOOH, lipid hydroperoxides; Met(O), methionine sulfoxide; Met112, methionine 112 residue; Met86, methionine 86 residue; oxLDL, oxidized low

  15. Impact of the New Generation Reconstituted Surfactant CHF5633 on Human CD4+ Lymphocytes.

    Directory of Open Access Journals (Sweden)

    Markus Fehrholz

    Full Text Available Natural surfactant preparations, commonly isolated from porcine or bovine lungs, are used to treat respiratory distress syndrome in preterm infants. Besides biophysical effectiveness, several studies have documented additional immunomodulatory properties. Within the near future, synthetic surfactant preparations may be a promising alternative. CHF5633 is a new generation reconstituted synthetic surfactant preparation with defined composition, containing dipalmitoyl-phosphatidylcholine, palmitoyl-oleoyl-phosphatidylglycerol and synthetic analogs of surfactant protein (SP- B and SP-C. While its biophysical effectiveness has been demonstrated in vitro and in vivo, possible immunomodulatory abilities are currently unknown.The aim of the current study was to define a potential impact of CHF5633 and its single components on pro- and anti-inflammatory cytokine responses in human CD4+ lymphocytes.Purified human CD4+ T cells were activated using anti CD3/CD28 antibodies and exposed to CHF5633, its components, or to the well-known animal-derived surfactant Poractant alfa (Curosurf®. Proliferative response and cell viability were assessed using flow cytometry and a methylthiazolyldiphenyltetrazolium bromide colorimetric assay. The mRNA expression of IFNγ, IL-2, IL-17A, IL-22, IL-4, and IL-10 was measured by quantitative PCR, while intracellular protein expression was assessed by means of flow cytometry.Neither CHF5633 nor any of its phospholipid components with or without SP-B or SP-C analogs had any influence on proliferative ability and viability of CD4+ lymphocytes under the given conditions. IFNγ, IL-2, IL-17A, IL-22, IL-4, and IL-10 mRNA as well as IFNγ, IL-2, IL-4 and IL-10 protein levels were unaffected in both non-activated and activated CD4+ lymphocytes after exposure to CHF5633 or its constituents compared to non-exposed controls. However, in comparison to Curosurf®, expression levels of anti-inflammatory IL-4 and IL-10 mRNA were

  16. Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane

    Energy Technology Data Exchange (ETDEWEB)

    BARCELLOS-HOFF, M. H; AGGELER, J.; RAM, T. G; BISSELL, M. J

    1989-02-01

    An essential feature of mammary gland differentiation during pregnancy is the formation of alveoli composed of polarized epithelial cells, which, under the influence of lactogenic hormones, secrete vectorially and sequester milk proteins. Previous culture studies have described either organization of cells polarized towards lumina containing little or no demonstrable tissue-specific protein, or establishment of functional secretory cells exhibiting little or no glandular architecture. In this paper, we report that tissue-specific vectorial secretion coincides with the formation of functional alveoli-like structures by primary mammary epithelial cells cultured on a reconstituted basement membrane matrix (derived from Engelbreth-Holm-Swarm murine tumour). Morphogenesis of these unique three-dimensional structures was initiated by cell-directed remodelling of the exogenous matrix leading to reorganization of cells into matrixensheathed aggregates by 24 h after plating. The aggregates subsequently cavitated, so that by day 6 the cells were organized into hollow spheres in which apical cell surfaces faced lumina sealed by tight junctions and basal surfaces were surrounded by a distinct basal lamina. The profiles of proteins secreted into the apical (luminal) and basal (medium) compartments indicated that these alveoli-like structures were capable of an appreciable amount of vectorial secretion. Immunoprecipitation with a broad spectrum milk antiserum showed that more than 80% of caseins were secreted into the lumina, whereas iron-binding proteins (both lactoferrin and transferrin) were present in comparable amounts in each compartment. Thus, these mammary cells established protein targeting pathways directing milk-specific proteins to the luminal compartment. A time course monitoring secretory activity demonstrated that establishment of tissue-specific vectorial secretion and increased total and milk protein secretion coincided with functional alveolar

  17. Contribution of the PET/CT with F.D.G. to evaluate the tri therapy and the lymphocyte reconstitution in patients infected by HIV; Apport de la TEP/TDM au FDG pour l'evaluation de la tritherapie et de la reconstitution lymphocytaire chez les patients infectes par le VIH

    Energy Technology Data Exchange (ETDEWEB)

    Hamza, F.; Yeddes, I.; El Bez, I.; Evangelista, E.; Meignan, M.; Itti, E. [CHU Henri-Mondor, Service de medecine nucleaire, 94 - Creteil (France); Lelievre, J.D.; Levy, Y. [CHU Henri-Mondor, service immunologie clinique, 94 - Creteil (France); Cheynier, R. [Institut Pasteur, service de virologie, 75 - Paris (France)

    2010-07-01

    Purpose: HIV infection is accompanied by impairment of lymphoid organs that can be detected by PET / CT with F.D.G.. The purpose of this study was to correlate lymphocyte reconstitution with the metabolic activity of lymphoid organs. Conclusions: PET / CT with F.D.G. is an appropriate assessment tool to evaluate the effect of antiviral treatment in lymphoid organs. In addition, she is helping to understand the mechanisms of lymphocyte reconstitution under triple therapy. (N.C.)

  18. The Abbey Theatre Digital Archive: a digitization project with dramatic impact

    Directory of Open Access Journals (Sweden)

    John Cox

    2017-11-01

    Full Text Available National University of Ireland Galway digitized the archive of the Abbey Theatre between 2012 and 2015. This was the largest theatre archive digitization project worldwide and it has had a major impact on the University and its Library. The scale of the digitization project presented a series of challenges, including fragile material, limited time, streamlined workflows, complex digital rights management and effective systems. The project was completed on time and on budget in 2015, using a ‘more product, less process’ approach. Access to the Abbey Theatre Digital Archive has delivered strong academic impact for the University, generating new research income and international connections as well as contributing to improved institutional ranking. The Digital Archive enables new types of research, including text and data mining, and has reshaped undergraduate curricula. It has also had a transformative effect on the Library as leader of the project. The role of the archivist has changed and partnerships with the academic community have strengthened. A growing emphasis on digital publication has been a catalyst for a function- rather than subject-based organizational structure which promotes participation in digital scholarship initiatives, with archives and special collections occupying a new position of prominence.

  19. Dramatic course of osteomyelitis in a patient treated with immediately placed dental implants suffering from uncontrolled diabetes: A case report.

    Science.gov (United States)

    Doll, Christian; Hartwig, Stefan; Nack, Claudia; Nahles, Susanne; Nelson, Katja; Raguse, Jan-Dirk

    2015-01-01

    To report a rare and dramatic complication following immediate dental implant placement in a heavy smoker, who had a delayed diagnosis of uncontrolled diabetes. In this case report we present the dramatic course of a 64-year old female patient treated with five immediate post-extractive dental implants in the mandible, who developed osteomyelitis, which manifested initially as local peri-implant inflammation and progressed into a spontaneous jaw fracture, despite repeated surgical interventions and antibiotic courses over a 3-year period, until diabetes was diagnosed. A symptom-free status could be achieved only after partial mandibulectomy, treatment of diabetes and reconstruction with a microvascular fibula free flap. In the presence of mandibular osteomyelitis refractory to therapy, yet undiagnosed underlying pathologies, such as diabetes, should be investigated and treated urgently.

  20. Dramatic inversion of charge polarity in diketopyrrolopyrrole-based organic field-effect transistors via a simple nitrile group substitution.

    Science.gov (United States)

    Yun, Hui-Jun; Kang, Seok-Ju; Xu, Yong; Kim, Seul Ong; Kim, Yun-Hi; Noh, Yong-Young; Kwon, Soon-Ki

    2014-11-19

    A record-breaking high electron mobility of 7.0 cm(2) V(-1) s(-1) for n-channel polymer OFETs is reported. By the incorporation of only one nitrile group as an electron-withdrawing function in the vinyl linkage of the DPP-based copolymer, a dramatic inversion of majority charge-carriers from holes to electrons is achieved. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Dramatic Rewritings of the Spanish Golden Age Theater of Cervantes´s La fuerza de la sangre

    Directory of Open Access Journals (Sweden)

    Juan Manuel Escudero Baztán

    2013-12-01

    Full Text Available This paper analyzes the Golden age spanish theater recreations of Cervantes’s exemplary novel La fuerza de la sangre. Specifically, the paper reviews three important stages in these recreations: La fuerza de la sangre of Guillen de Castro, El agravio satisfecho de Castillo Solórzano, and No hay cosa como callar de Calderon de la Barca. Different rewrites indicate a close relationship between the three dramatic texts through intertextuality and other influences.

  2. Surveillance, Auditing, and Feedback Can Reduce Surgical Site Infection Dramatically: Toward Zero Surgical Site Infection.

    Science.gov (United States)

    Manivannan, Bhavani; Gowda, Deepak; Bulagonda, Pradeep; Rao, Abhishek; Raman, Sai Suguna; Natarajan, Shanmuga Vadivoo

    2018-04-01

    We evaluated the Surveillance of Surgical Site Infection (SSI), Auditing, and Feedback (SAF) effect on the rate of compliance with an SSI care bundle and measured its effectiveness in reducing the SSI rate. A prospective cohort study from January 2014 to December 2016 was classified into three phases: pre-SAF, early-SAF, and late-SAF. Pre-operative baseline characteristics of 24,677 patients who underwent orthopedic, cardiovascular thoracic surgery (CTVS) or urologic operations were recorded. Univariable analyses of the SSI rates in the pre-SAF and post-SAF phases were performed. Percentage compliance and non-compliance with each care component were calculated. Correlation between reduction in the SSI rate and increase in compliance with the pre-operative, peri-operative, and post-operative care-bundle components was performed using the Spearman test. There was a significant decrease in the SSI rate in orthopedic procedures that involved surgical implantation and in mitral valve/aortic valve (MVR/AVR) cardiac operations, with a relative risk (RR) ratio of 0.19 (95% confidence interval [CI] 0.12-0.31) and 0.08 (95% CI 0.03-0.22), respectively. The SSI rate was inversely correlated with the rate of compliance with pre-operative (r = -0.738; p = 0.037), peri-operative (r = - 0.802; p = 0.017), and post-operative (r = -0.762; p = 0.028) care bundles. Implementation of the Surveillance of SSI, Auditing, and Feedback bundle had a profound beneficial effect on the SSI rate, thereby reducing healthcare costs and improving patient quality of life.

  3. Virtual historical reconstitution of the main altarpiece of the Espírito Santo Church, in Évora: application of web-based infographics to Cultural Heritage

    Directory of Open Access Journals (Sweden)

    Catarina Pereira

    2016-01-01

    Full Text Available As a part of a multidisciplinary and integrated research, including conservation sciences and history, a proposal is presented for the historical reconstitution and the virtual restoration of the mannerist altarpiece of the main altar at the Espírito Santo Church, in Évora. The collected data is abundant and the scientific information, because of its technicality, is less prone to be easily understood by the general public, thus becoming less accessible. Web-based infographics are explored as privileged forms of disseminating results and raising awareness to Cultural Heritage. The project materializes as an Internet platform where data and a reconstitution proposal are shared in a visual and interactive way. In addition to the digital virtual reconstitution (2D, some tridimensional models (3D are presented of various elements of the altarpiece, obtained using methods of computer graphics and digital photogrammetry.

  4. Growth and characterization of different human rhinovirus C types in three-dimensional human airway epithelia reconstituted in vitro

    International Nuclear Information System (INIS)

    Tapparel, Caroline; Sobo, Komla; Constant, Samuel; Huang, Song; Van Belle, Sandra; Kaiser, Laurent

    2013-01-01

    New molecular diagnostic tools have recently allowed the discovery of human rhinovirus species C (HRV-C) that may be overrepresented in children with lower respiratory tract complications. Unlike HRV-A and HRV-B, HRV-C cannot be propagated in conventional immortalized cell lines and their biological properties have been difficult to study. Recent studies have described the successful amplification of HRV-C15, HRV-C11, and HRV-C41 in sinus mucosal organ cultures and in fully differentiated human airway epithelial cells. Consistent with these studies, we report that a panel of clinical HRV-C specimens including HRV-C2, HRV-C7, HRV-C12, HRV-C15, and HRV-C29 types were all capable of mediating productive infection in reconstituted 3D human primary upper airway epithelial tissues and that the virions enter and exit preferentially through the apical surface. Similar to HRV-A and HRV-B, our data support the acid sensitivity of HRV-C. We observed also that the optimum temperature requirement during HRV-C growth may be type-dependent. - Highlights: • A 3D human upper airway epithelia reconstituted in vitro supports HRV-C growth. • HRV-Cs enter and exit preferentially at the apical side of this ALI culture system. • HRV-Cs are acid sensitive. • Temperature sensitivity may be type-dependent for HRV-Cs

  5. Photoactive protochlorophyllide-enzyme complexes reconstituted with PORA, PORB and PORC proteins of A. thaliana: fluorescence and catalytic properties.

    Directory of Open Access Journals (Sweden)

    Michał Gabruk

    Full Text Available Photoactive Pchlide-POR-NADPH complexes were reconstituted using protochlorophyllide (Pchlide and recombinant light-dependent protochlorophyllide oxidoreductase (POR proteins, His₆-PORA, His₆-PORB and His₆-PORC, from Arabidopsis thaliana. We did not observe any differences in the kinetics of the protochlorophyllide photoreduction at room temperature among the PORA, PORB and PORC proteins. In contrast, the PORC protein showed lower yield of Chlide formation than PORA and PORB when preincubated in the dark for 30 min and then illuminated for a short time. The most significant observation was that reconstituted Pchlide-POR-NADPH complexes showed fluorescence maxima at 77 K similar to those observed for highly aggregated Pchlide-POR-NADPH complexes in prolamellar bodies (PLBs in vivo. Homology models of PORA, PORB and PORC of Arabidopsis thaliana were developed to compare predicted structures of POR isoforms. There were only slight structural differences, mainly in the organisation of helices and loops, but not in the shape of whole molecules. This is the first comparative analysis of all POR isoforms functioning at different stages of A. thaliana development.

  6. Reconstituted injectable hyaluronic acid: expanded applications in facial aesthetics and additional thoughts on the mechanism of action in cosmetic medicine.

    Science.gov (United States)

    Fagien, Steven; Cassuto, Daniel

    2012-07-01

    As we currently have a better understanding of the components of facial aging, injectable filling agents have a larger role in nonsurgical facial rejuvenation. Many products are now available worldwide as options for treating the spectrum of indications that include particular agents used for the treatment of various facial rhytides and in larger quantities for regional or panfacial volumization. The latter indication has been most popularly treated with a host of agents classified as hyaluronans or injectable hyaluronic acid gels. The former had been traditionally and more successfully treated with injectable collagen or dermal matrix agents. Attempts to confuse these generalized approaches have often led to unsatisfactory results. With the precipitous disappearance of the availability of most injectable collagen substances, other methods of treating finer facial rhytides have been explored. Of the many options available for the treatment of facial fine lines, the authors describe a novel technique of formulating reduced concentrations of existing hyaluronic acid gels by variable dilution/reconstitution. Over 350 of the authors' collective patients have been treated with this approach. The results have been highly satisfactory, with greater persistence, and have challenged traditional concepts of mechanism of action and biodegradation and suggest alternative options in formulations of currently available products. Reconstituted injectable hyaluronic acid is a highly effective and viable alternative for the aesthetic treatment of facial fine lines and other components of aging. Observations, treatment guidelines, and a rationale for this novel use and concepts to explain these finding are contained in this article.

  7. Growth and characterization of different human rhinovirus C types in three-dimensional human airway epithelia reconstituted in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Tapparel, Caroline, E-mail: Caroline.Tapparel@hcuge.ch [Laboratory of Virology, Division of Infectious Diseases and Division of Laboratory Medicine, University of Geneva Hospitals, 4 Rue Gabrielle-Perret-Gentil, 1211 Geneva 14 (Switzerland); Sobo, Komla [Laboratory of Virology, Division of Infectious Diseases and Division of Laboratory Medicine, University of Geneva Hospitals, 4 Rue Gabrielle-Perret-Gentil, 1211 Geneva 14 (Switzerland); Constant, Samuel; Huang, Song [Epithelix sárl, 14 Chemin des Aulx, 1228 Plan les Ouates, Geneva (Switzerland); Van Belle, Sandra; Kaiser, Laurent [Laboratory of Virology, Division of Infectious Diseases and Division of Laboratory Medicine, University of Geneva Hospitals, 4 Rue Gabrielle-Perret-Gentil, 1211 Geneva 14 (Switzerland)

    2013-11-15

    New molecular diagnostic tools have recently allowed the discovery of human rhinovirus species C (HRV-C) that may be overrepresented in children with lower respiratory tract complications. Unlike HRV-A and HRV-B, HRV-C cannot be propagated in conventional immortalized cell lines and their biological properties have been difficult to study. Recent studies have described the successful amplification of HRV-C15, HRV-C11, and HRV-C41 in sinus mucosal organ cultures and in fully differentiated human airway epithelial cells. Consistent with these studies, we report that a panel of clinical HRV-C specimens including HRV-C2, HRV-C7, HRV-C12, HRV-C15, and HRV-C29 types were all capable of mediating productive infection in reconstituted 3D human primary upper airway epithelial tissues and that the virions enter and exit preferentially through the apical surface. Similar to HRV-A and HRV-B, our data support the acid sensitivity of HRV-C. We observed also that the optimum temperature requirement during HRV-C growth may be type-dependent. - Highlights: • A 3D human upper airway epithelia reconstituted in vitro supports HRV-C growth. • HRV-Cs enter and exit preferentially at the apical side of this ALI culture system. • HRV-Cs are acid sensitive. • Temperature sensitivity may be type-dependent for HRV-Cs.

  8. Promotion of hair follicle development and trichogenesis by Wnt-10b in cultured embryonic skin and in reconstituted skin

    International Nuclear Information System (INIS)

    Ouji, Yukiteru; Yoshikawa, Masahide; Shiroi, Akira; Ishizaka, Shigeaki

    2006-01-01

    We previously showed that Wnt-10b promoted the differentiation of primary skin epithelial cells (MPSEC) toward hair shaft and inner root sheath of the hair follicle (IRS) cells in vitro. In the present study, we found that Wnt-10b promotes the development of hair follicles using a culture of mouse embryonic skin tissue and trichogenesis using a reconstitution experiment with nude mice. Hair follicle development was observed in skin taken from mouse embryos on embryonic day 10.5 following a 2-day culture with recombinant Wnt-10b (rWnt-10b), however, not without rWnt-10b. Brown hair growth was observed at the site of reconstituted skin in Balb/c nude mice where dermal fibroblasts and keratinocytes, derived from C3H/HeN new born mice, were transplanted with Wnt-10b-producing COS cells (Wnt-COS). Without the co-transplantation of Wnt-COS, no hair growth was observed. Our results suggest an important role of Wnt-10b in the initiation of hair follicle development and following trichogenesis

  9. Thymectomized, irradiated, and bone marrow-reconstituted chimeras have normal cytolytic T lymphocyte precursors but a defect in lymphokine production

    International Nuclear Information System (INIS)

    Duprez, V.; Maziarz, R.; Weinberger, O.; Burakoff, S.J.

    1984-01-01

    A model system has been developed to study extrathymic T cell differentiation; mice have been thymectomized, lethally irradiated, and reconstituted with bone marrow cells depleted of Thy-1 + cells. After 8 wk, the spleen cells of these athymic, bone marrow-reconstituted chimeras contain Thy-1 + precytolytic T lymphocytes (CTL) that are able to respond to antigen only if supernatant from Con A-activated T cells is added to culture. The phenotype of these pre-CTL is similar to that of thymocytes, suggesting that they may be immature T cells. Initial evaluation of the CTL repertoire of these athymic mice demonstrated that the CTL generated to trinitrophenyl-modified syngeneic cells are H-2-restricted, and that the CTL generated to alloantigens have many of the cross-reactivities observed in normal mice but not in nude mice. In this report, the authors demonstrate a helper T cell defect in these thymectomized chimeras. These chimeras lack an Ly-1 + helper cell required for thymocytes to differentiate to CTL. Further studies revealed that when spleen cells from these thymectomized chimeras were stimulated with Con A, they produced normal levels of interleukin 2. However, these splenocytes were defective in the production of another factor needed for CTL differentiation

  10. Critical role of CD4 T cells in maintaining lymphoid tissue structure for immune cell homeostasis and reconstitution.

    Science.gov (United States)

    Zeng, Ming; Paiardini, Mirko; Engram, Jessica C; Beilman, Greg J; Chipman, Jeffrey G; Schacker, Timothy W; Silvestri, Guido; Haase, Ashley T

    2012-08-30

    Loss of the fibroblastic reticular cell (FRC) network in lymphoid tissues during HIV-1 infection has been shown to impair the survival of naive T cells and limit immune reconstitution after antiretroviral therapy. What causes this FRC loss is unknown. Because FRC loss correlates with loss of both naive CD4 and CD8 T-cell subsets and decreased lymphotoxin-β, a key factor for maintenance of FRC network, we hypothesized that loss of naive T cells is responsible for loss of the FRC network. To test this hypothesis, we assessed the consequences of antibody-mediated depletion of CD4 and CD8 T cells in rhesus macaques and sooty mangabeys. We found that only CD4 T-cell depletion resulted in FRC loss in both species and that this loss was caused by decreased lymphotoxin-β mainly produced by the CD4 T cells. We further found the same dependence of the FRC network on CD4 T cells in HIV-1-infected patients before and after antiretroviral therapy and in other immunodeficiency conditions, such as CD4 depletion in cancer patients induced by chemotherapy and irradiation. CD4 T cells thus play a central role in the maintenance of lymphoid tissue structure necessary for their own homeostasis and reconstitution.

  11. Purification, Reconstitution, and Inhibition of Cytochrome P-450 Sterol Δ22-Desaturase from the Pathogenic Fungus Candida glabrata

    Science.gov (United States)

    Lamb, David C.; Maspahy, Segula; Kelly, Diane E.; Manning, Nigel J.; Geber, Antonia; Bennett, John E.; Kelly, Steven L.

    1999-01-01

    Sterol Δ22-desaturase has been purified from a strain of Candida glabrata with a disruption in the gene encoding sterol 14α-demethylase (cytochrome P-45051; CYP51). The purified cytochrome P-450 exhibited sterol Δ22-desaturase activity in a reconstituted system with NADPH–cytochrome P-450 reductase in dilaurylphosphatidylcholine, with the enzyme kinetic studies revealing a Km for ergosta-5,7-dienol of 12.5 μM and a Vmax of 0.59 nmol of this substrate metabolized/min/nmol of P-450. This enzyme is encoded by CYP61 (ERG5) in Saccharomyces cerevisiae, and homologues have been shown in the Candida albicans and Schizosaccharomyces pombe genome projects. Ketoconazole, itraconazole, and fluconazole formed low-spin complexes with the ferric cytochrome and exhibited type II spectra, which are indicative of an interaction between the azole moiety and the cytochrome heme. The azole antifungal compounds inhibited reconstituted sterol Δ22-desaturase activity by binding to the cytochrome with a one-to-one stoichiometry, with total inhibition of enzyme activity occurring when equimolar amounts of azole and cytochrome P-450 were added. These results reveal the potential for sterol Δ22-desaturase to be an antifungal target and to contribute to the binding of drugs within the fungal cell. PMID:10390230

  12. Efficient purification and reconstitution of ATP binding cassette transporter B6 (ABCB6) for functional and structural studies.

    Science.gov (United States)

    Chavan, Hemantkumar; Khan, Mohiuddin Md Taimur; Tegos, George; Krishnamurthy, Partha

    2013-08-02

    The mitochondrial ATP binding cassette transporter ABCB6 has been associated with a broad range of physiological functions, including growth and development, therapy-related drug resistance, and the new blood group system Langereis. ABCB6 has been proposed to regulate heme synthesis by shuttling coproporphyrinogen III from the cytoplasm into the mitochondria. However, direct functional information of the transport complex is not known. To understand the role of ABCB6 in mitochondrial transport, we developed an in vitro system with pure and active protein. ABCB6 overexpressed in HEK293 cells was solubilized from mitochondrial membranes and purified to homogeneity. Purified ABCB6 showed a high binding affinity for MgATP (Kd = 0.18 μM) and an ATPase activity with a Km of 0.99 mM. Reconstitution of ABCB6 into liposomes allowed biochemical characterization of the ATPase including (i) substrate-stimulated ATPase activity, (ii) transport kinetics of its proposed endogenous substrate coproporphyrinogen III, and (iii) transport kinetics of substrates identified using a high throughput screening assay. Mutagenesis of the conserved lysine to alanine (K629A) in the Walker A motif abolished ATP hydrolysis and substrate transport. These results suggest a direct interaction between mitochondrial ABCB6 and its transport substrates that is critical for the activity of the transporter. Furthermore, the simple immunoaffinity purification of ABCB6 to near homogeneity and efficient reconstitution of ABCB6 into liposomes might provide the basis for future studies on the structure/function of ABCB6.

  13. Droplet interface bilayer reconstitution and activity measurement of the mechanosensitive channel of large conductance from Escherichia coli.

    Science.gov (United States)

    Barriga, Hanna M G; Booth, Paula; Haylock, Stuart; Bazin, Richard; Templer, Richard H; Ces, Oscar

    2014-09-06

    Droplet interface bilayers (DIBs) provide an exciting new platform for the study of membrane proteins in stable bilayers of controlled composition. To date, the successful reconstitution and activity measurement of membrane proteins in DIBs has relied on the use of the synthetic lipid 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC). We report the functional reconstitution of the mechanosensitive channel of large conductance (MscL) into DIBs composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), a lipid of significantly greater biological relevance than DPhPC. MscL functionality has been demonstrated using a fluorescence-based assay, showing that dye flow occurs across the DIB when MscL is gated by the cysteine reactive chemical 2-(trimethylammonium)ethyl methane thiosulfonate bromide (MTSET). MscL has already been the subject of a number of studies investigating its interaction with the membrane. We propose that this method will pave the way for future MscL studies looking in detail at the effects of controlled composition or membrane asymmetry on MscL activity using biologically relevant lipids and will also be applicable to other lipid-protein systems, paving the way for the study of membrane proteins in DIBs with biologically relevant lipids.

  14. Metabolism of benzene and phenol by a reconstituted purified phenobarbital induced rat liver mixed function oxidase system

    International Nuclear Information System (INIS)

    Griffiths, J.C.

    1986-01-01

    Cytochrome P-450 and the electron-donor, NADPH-cytochrome c reductase were isolated from phenobarbital induced rat liver microsomes. Both benzene and its primary metabolite phenol, were substrates for the reconstituted purified phenobarbital induced rat liver mixed function oxidase system. Benzene was metabolized to phenol and the polyhydroxylated metabolites; catechol, hydroquinone and 1,2,4 benzenetriol. Benzene elicited a Type I spectral change upon its interaction with the cytochrome P-450 while phenol's interaction with the cytochrome P-450 produced a reverse Type I spectra. The formation of phenol showed a pH optimum of 7.0 compared with 6.6-6.8 for the production of the polyhyrdoxylated metabolites. Cytochrome P-450 inhibitors, such as metyrapone and SKF 525A, diminished the production of phenol from benzene but not the production of the polyhydroxylated metabolites from phenol. The radical trapping agents, DMSO, KTBA and mannitol, decreased the recovery of polyhydroxylated metabolites, from 14 C-labeled benzene and/or phenol. As KTBA and DMSO interacted with OH. There was a concomitant release of ethylene and methane, which was measured. Desferrioxamine, an iron-chelator and catalase also depressed the recovery of polyhydroxylated metabolites. In summary, benzene and phenol were both substrates for this reconstituted purified enzyme system, but they differed in binding to cytochrome P-450, pH optima and mode of hydroxylation

  15. Effect of radiation on reconstitution of skin equivalent (dermal alterations); Effet de l`irradiation sur la reconstruction d`une peau equivalente (alterations dermiques)

    Energy Technology Data Exchange (ETDEWEB)

    Gentilhomme, E.; Bergier, J.; Richard, M.; Neveux, Y.

    1994-12-31

    Dermal equivalents have been treated by single doses of gamma irradiation of 10, 20, 30 and 50 Gray. Numerations at different times show a dose and time dependant diminution of cellular population. This diminution is histologically observed in dermal part of reconstituted skin, in association with cellular and functional alterations of fibroblast cells. Modifications of epidermal epithelia are also noted in some reconstituted skin. This model would be useful to apprehend the effect of a dermal irradiation lesion on the later epidermization. (author). 4 refs.

  16. Analysis of some optical properties of a native and reconstituted photosystem II antenna complex, CP29 : Pigment binding sites can be occupied by chlorophyll a or chlorophyll b and determine spectral forms

    NARCIS (Netherlands)

    Giuffra, Elisabetta; Zucchelli, Giuseppe; Sandona, Dorianna; Croce, Roberta; Cugini, Daniela; Garlaschi, Flavio M.; Bassi, Roberto; Jennings, Robert C.

    1997-01-01

    The minor photosystem II antenna complex CP29(Lhcb-4) has been reconstituted in vitro with the Lhcb-4 apoprotein, overexpressed in Escherichia coli, and the native pigments. Modulation of the pigment composition during reconstitution yields binding products with markedly different chlorophyll a/b

  17. Chekhov’s dramatic art and cinematograph: implementation of author’s assumption in active shot mode

    OpenAIRE

    Bezrukov Andrei Nikolayevich

    2017-01-01

    The article analyses the art specificity of Chekhov’s legacy in dramatic art based on implementing in national film industry. Chekhov’s drama is divertive in its interpretation by the reader, the researcher, and in the director’s cut. Eccentricity frequently occurs in his dramaturgy. In literary text this device is not only a part of aesthetic picturing but also the form of overthinking, transformation of reality and changing the attitude to reality. The main division of the text form alters ...

  18. Dramatic Influence of an Anionic Donor on the Oxygen-Atom Transfer Reactivity of a MnV–Oxo Complex

    Science.gov (United States)

    Neu, Heather M; Quesne, Matthew G; Yang, Tzuhsiung; Prokop-Prigge, Katharine A; Lancaster, Kyle M; Donohoe, James; DeBeer, Serena; de Visser, Sam P; Goldberg, David P

    2014-01-01

    Addition of an anionic donor to an MnV(O) porphyrinoid complex causes a dramatic increase in 2-electron oxygen-atom-transfer (OAT) chemistry. The 6-coordinate [MnV(O)(TBP8Cz)(CN)]− was generated from addition of Bu4N+CN− to the 5-coordinate MnV(O) precursor. The cyanide-ligated complex was characterized for the first time by Mn K-edge X-ray absorption spectroscopy (XAS) and gives Mn–O=1.53 Å, Mn–CN=2.21 Å. In combination with computational studies these distances were shown to correlate with a singlet ground state. Reaction of the CN− complex with thioethers results in OAT to give the corresponding sulfoxide and a 2e−-reduced MnIII(CN)− complex. Kinetic measurements reveal a dramatic rate enhancement for OAT of approximately 24 000-fold versus the same reaction for the parent 5-coordinate complex. An Eyring analysis gives ΔH≠=14 kcal mol−1, ΔS≠=−10 cal mol−1 K−1. Computational studies fully support the structures, spin states, and relative reactivity of the 5- and 6-coordinate MnV(O) complexes. PMID:25256417

  19. Fasting, but Not Aging, Dramatically Alters the Redox Status of Cysteine Residues on Proteins in Drosophila melanogaster

    Directory of Open Access Journals (Sweden)

    Katja E. Menger

    2015-06-01

    Full Text Available Altering the redox state of cysteine residues on protein surfaces is an important response to environmental challenges. Although aging and fasting alter many redox processes, the role of cysteine residues is uncertain. To address this, we used a redox proteomic technique, oxidative isotope-coded affinity tags (OxICAT, to assess cysteine-residue redox changes in Drosophila melanogaster during aging and fasting. This approach enabled us to simultaneously identify and quantify the redox state of several hundred cysteine residues in vivo. Cysteine residues within young flies had a bimodal distribution with peaks at ∼10% and ∼85% reversibly oxidized. Surprisingly, these cysteine residues did not become more oxidized with age. In contrast, 24 hr of fasting dramatically oxidized cysteine residues that were reduced under fed conditions while also reducing cysteine residues that were initially oxidized. We conclude that fasting, but not aging, dramatically alters cysteine-residue redox status in D. melanogaster.

  20. Tensile Mechanical Properties and Dynamic Collagen Fiber Re-Alignment of the Murine Cervix are Dramatically Altered Throughout Pregnancy.

    Science.gov (United States)

    Barnum, Carrie E; Fey, Jennifer L; Weiss, Stephanie N; Barila, Guillermo; Brown, Amy G; Connizzo, Brianne K; Shetye, Snehal S; Elovitz, Michal A; Soslowsky, Louis J

    2017-06-01

    The cervix is a unique organ able to dramatically change its shape and function by serving as a physical barrier for the growing fetus and then undergoing dramatic dilation allowing for delivery of a term infant. As a result, the cervix endures changing mechanical forces from the growing fetus. There is an emerging concept that the cervix may change or remodel "early" in many cases of spontaneous preterm birth (sPTB). However, the mechanical role of the cervix in both normal and preterm birth remains unclear. Therefore, the primary objective of this study was to determine the mechanical and structural responses of murine cervical tissue throughout a normal gestational time course. In this study, both tissue structural and material properties were determined via a quasi-static tensile load-to-failure test, while simultaneously obtaining dynamic collagen fiber re-alignment via cross-polarization imaging. This study demonstrated that the majority of the mechanical properties evaluated decreased at midgestation and not just at term, while collagen fiber re-alignment occurred earlier in the loading curve for cervices at term. This suggests that although structural changes in the cervix occur throughout gestation, the differences in material properties function in combination with collagen fiber re-alignment as mechanical precursors to regulate term gestation. This work lays a foundation for investigating cervical biomechanics and the role of the cervix in preterm birth.