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Sample records for activities form g-646

  1. 75 FR 5097 - Agency Information Collection Activities: Form G-646, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-01

    ... Information Collection Under Review: Form G- 646, Sworn Statement of Refugee Applying for Admission to the... approved information collection. (2) Title of the Form/Collection: Sworn Statement of Refugee Applying for... eligibility for the admission of the applicants to the United states as refugees. (5) An estimate of the...

  2. 76 FR 11807 - Agency Information Collection Activities: Form G-646, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-03

    ... Information Collection Under Review: Form G- 646, Sworn Statement of Refugee Applying for Admission to the...) Title of the Form/Collection: Sworn Statement of Refugee Applying for Admission to the United States. (3... the applicants to the United States as refugees. (5) An estimate of the total number of...

  3. 77 FR 33760 - Agency Information Collection Activities: Form G-646, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-07

    ... Information Collection Under Review: Form G- 646, Sworn Statement of Refugee Applying for Admission to the... currently approved information collection. (2) Title of the Form/Collection: Sworn Statement of Refugee... eligibility for the admission of the applicants to the United States as refugees. (5) An estimate of the...

  4. 76 FR 61725 - Agency Information Collection Activities: Case Submission Form, Case Assistance Form; (Form DHS...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-05

    ... SECURITY Agency Information Collection Activities: Case Submission Form, Case Assistance Form; (Form DHS... CIS Ombudsman to identify the issue such as: (1) A case problem which is a request for information.... SUMMARY: The Department of Homeland Security, Office of the Citizenship and Immigration Service...

  5. Active superconducting devices formed of thin films

    DOEpatents

    Martens, Jon S.; Beyer, James B.; Nordman, James E.; Hohenwarter, Gert K. G.

    1991-05-28

    Active superconducting devices are formed of thin films of superconductor which include a main conduction channel which has an active weak link region. The weak link region is composed of an array of links of thin film superconductor spaced from one another by voids and selected in size and thickness such that magnetic flux can propagate across the weak link region when it is superconducting. Magnetic flux applied to the weak link region will propagate across the array of links causing localized loss of superconductivity in the links and changing the effective resistance across the links. The magnetic flux can be applied from a control line formed of a superconducting film deposited coplanar with the main conduction channel and weak link region on a substrate. The devices can be formed of any type to superconductor but are particularly well suited to the high temperature superconductors since the devices can be entirely formed from coplanar films with no overlying regions. The devices can be utilized for a variety of electrical components, including switching circuits, amplifiers, oscillators and modulators, and are well suited to microwave frequency applications.

  6. 75 FR 78263 - Agency Information Collection Activities: Extension of an Existing Information Collection...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-15

    ... under review; Form G- 646, Sworn Statement of Refugee Applying for Admission to the United States; OMB... Refugee Applying for Admission to the United States. (3) Agency form number, if any, and the applicable... determine eligibility for the admission of the applicants to the United States as refugees. (5) An...

  7. 77 FR 14407 - Agency Information Collection Activities: Extension of an Existing Information Collection...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-09

    ... under review; Form G- 646, Sworn Statement of Refugee Applying for Admission to the United States. The...: Sworn Statement of Refugee Applying for Admission to the United States. (3) Agency form number, if any... States as refugees. (5) An estimate of the total number of respondents and the amount of time...

  8. 78 FR 57870 - Agency Information Collection Activities: Registration for Classification as Refugee; Revision of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-20

    ... for Classification as Refugee; Revision of a Currently Approved Collection ACTION: 60-day notice...-590 provides a uniform method for applicants to apply for refugee status and contains the information... that have been transferred from Form G-646, Sworn Statement of Refugee Applying for Admission into...

  9. Active compounds release from semisolid dosage forms.

    PubMed

    Olejnik, Anna; Goscianska, Joanna; Nowak, Izabela

    2012-11-01

    The aim of this paper is to review all the aspects of the in vitro release testing (IVRT) from semisolid dosage forms. Although none of the official dissolution methods has been specified for use with semisolid dosage forms, their utility for assessing release rates of drugs from semisolid dosage forms has become a topic of considerable interest. One can expect to overcome such complexity in the future, when the official "Topical and Transdermal Drug Products-Product Performance Tests" will be published in an issue of the Pharmacopeial Forum. Many factors such as type of the dissolution medium, membrane, temperature, and speed have an influence on the mechanism and kinetics of the release testing from gels, creams, and ointments; therefore, those parameters have been widely discussed.

  10. Neural network with formed dynamics of activity

    SciTech Connect

    Dunin-Barkovskii, V.L.; Osovets, N.B.

    1995-03-01

    The problem of developing a neural network with a given pattern of the state sequence is considered. A neural network structure and an algorithm, of forming its bond matrix which lead to an approximate but robust solution of the problem are proposed and discussed. Limiting characteristics of the serviceability of the proposed structure are studied. Various methods of visualizing dynamic processes in a neural network are compared. Possible applications of the results obtained for interpretation of neurophysiological data and in neuroinformatics systems are discussed.

  11. 76 FR 42129 - Agency Information Collection Activities: Case Submission Form, Case Assistance Form

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-18

    ... SECURITY Agency Information Collection Activities: Case Submission Form, Case Assistance Form (Form DHS... the CIS Ombudsman to identify the issue such as: (1) A case problem which is a request for information.... SUMMARY: The Department of Homeland Security, Office of the Citizenship and Immigration Service...

  12. Pu-238 fuel form activities, January 1-31, 1982

    SciTech Connect

    Not Available

    1982-03-01

    This monthly report for /sup 238/Pu fuel form activities has two main sections: SRP-PuFF facility and SRL fuel form activities. The program status, budget information, and milestone schedules are discussed in each main section. The Work Breakdown Structure (WBS) for this program is shown. Only one monthly report per year is processed for EDB.

  13. New Forms of Student Activism: Lobbying, Trusteeing, and Collective Bargaining.

    ERIC Educational Resources Information Center

    Beeler, Kent D.

    1979-01-01

    This article focuses on three new forms of student activism: lobbying, trusteeing, and collective bargaining. Related aspects of student involvement in the political, legal, and consumer areas are discussed briefly. (Author)

  14. Pu-238 fuel form activities, June 1-30, 1980

    SciTech Connect

    Not Available

    1980-07-18

    This monthly report for Pu-238 Fuel Form Activities has two main sections: SRP-PuFF Pu-238 Fuel Form Production Processes and SRL Pu-238 Fuel Form Research and Development. The program status, budget information, and milestone information are discussed in each main section. The Work Breakdown Structures (WBS) for this program is outlined. Only one monthly report per year is processed for EDB.

  15. Form-Focused Discovery Activities in English Classes

    ERIC Educational Resources Information Center

    Ogeyik, Muhlise Cosgun

    2011-01-01

    Form-focused discovery activities allow language learners to grasp various aspects of a target language by contributing implicit knowledge by using discovered explicit knowledge. Moreover, such activities can assist learners to perceive and discover the features of their language input. In foreign language teaching environments, they can be used…

  16. Activated carbon fibers and engineered forms from renewable resources

    DOEpatents

    Baker, Frederick S.

    2010-06-01

    A method of producing activated carbon fibers (ACFs) includes the steps of providing a natural carbonaceous precursor fiber material, blending the carbonaceous precursor material with a chemical activation agent to form chemical agent-impregnated precursor fibers, spinning the chemical agent-impregnated precursor material into fibers, and thermally treating the chemical agent-impregnated precursor fibers. The carbonaceous precursor material is both carbonized and activated to form ACFs in a single step. The method produces ACFs exclusive of a step to isolate an intermediate carbon fiber.

  17. Activated carbon fibers and engineered forms from renewable resources

    DOEpatents

    Baker, Frederick S

    2013-02-19

    A method of producing activated carbon fibers (ACFs) includes the steps of providing a natural carbonaceous precursor fiber material, blending the carbonaceous precursor material with a chemical activation agent to form chemical agent-impregnated precursor fibers, spinning the chemical agent-impregnated precursor material into fibers, and thermally treating the chemical agent-impregnated precursor fibers. The carbonaceous precursor material is both carbonized and activated to form ACFs in a single step. The method produces ACFs exclusive of a step to isolate an intermediate carbon fiber.

  18. Pore-forming activity of clostridial binary toxins.

    PubMed

    Knapp, O; Benz, R; Popoff, M R

    2016-03-01

    Clostridial binary toxins (Clostridium perfringens Iota toxin, Clostridium difficile transferase, Clostridium spiroforme toxin, Clostridium botulinum C2 toxin) as Bacillus binary toxins, including Bacillus anthracis toxins consist of two independent proteins, one being the binding component which mediates the internalization into cell of the intracellularly active component. Clostridial binary toxins induce actin cytoskeleton disorganization through mono-ADP-ribosylation of globular actin and are responsible for enteric diseases. Clostridial and Bacillus binary toxins share structurally and functionally related binding components which recognize specific cell receptors, oligomerize, form pores in endocytic vesicle membrane, and mediate the transport of the enzymatic component into the cytosol. Binding components retain the global structure of pore-forming toxins (PFTs) from the cholesterol-dependent cytotoxin family such as perfringolysin. However, their pore-forming activity notably that of clostridial binding components is more related to that of heptameric PFT family including aerolysin and C. perfringens epsilon toxin. This review focuses upon pore-forming activity of clostridial binary toxins compared to other related PFTs. This article is part of a Special Issue entitled: Pore-Forming Toxins edited by Mauro Dalla Serra and Franco Gambale.

  19. Crystal Structure of an Active Form of Human MMP-1

    PubMed Central

    Iyer, Shalini; Visse, Robert; Nagase, Hideaki; Acharya, K. Ravi

    2006-01-01

    The extracellular matrix is a dynamic environment that constantly undergoes remodelling and degradation during vital physiological processes such as angiogenesis, wound healing, and development. Unbalanced extracellular matrix breakdown is associated with many diseases such as arthritis, cancer and fibrosis. Interstitial collagen is degraded by matrix metalloproteinases with collagenolytic activity by MMP-1, MMP-8 and MMP-13, collectively known as the collagenases. Matrix metalloproteinase 1 (MMP-1) plays a pivotal role in degradation of interstitial collagen types I, II, and III. Here, we report the crystal structure of the active form of human MMP-1 at 2.67 Å resolution. This is the first MMP-1 structure that is free of inhibitor and a water molecule essential for peptide hydrolysis is observed coordinated with the active site zinc. Comparing this structure with the human proMMP-1 shows significant structural differences, mainly in the relative orientation of the hemopexin domain, between the pro form and active form of the human enzyme. PMID:16890240

  20. Centrally formed acetaldehyde mediates ethanol-induced brain PKA activation.

    PubMed

    Tarragon, E; Baliño, P; Aragon, C M G

    2014-09-19

    Centrally formed acetaldehyde has proven to be responsible for several psychopharmacological effects induced by ethanol. In addition, it has been suggested that the cAMP-PKA signaling transduction pathway plays an important role in the modulation of several ethanol-induced behaviors. Therefore, we hypothesized that acetaldehyde might be ultimately responsible for the activation of this intracellular pathway. We used three pharmacological agents that modify acetaldehyde activity (α-lipoic acid, aminotriazole, and d-penicillamine) to study the role of this metabolite on EtOH-induced PKA activation in mice. Our results show that the injection of α-lipoic acid, aminotriazole and d-penicillamine prior to acute EtOH administration effectively blocks the PKA-enhanced response to EtOH in the brain. These results strongly support the hypothesis of a selective release of acetaldehyde-dependent Ca(2+) as the mechanism involved in the neurobehavioral effects elicited by EtOH.

  1. 76 FR 27077 - Agency Information Collection Activities: Form AR-11 and Form AR-11SR, Extension of an Existing...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-10

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form AR-11 and Form AR- 11SR, Extension of an Existing Information Collection; Comment Request ACTION: 60-Day Notice of Information Collection under Review: Form AR- 11 and Form AR-11SR, Alien's Change of Address...

  2. 76 FR 30738 - Agency Information Collection Activities: Form G-845 and Form G-845 Supplement, Revision of a...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-26

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form G-845 and Form G- 845 Supplement, Revision of a Currently Approved Information Collection; Comment Request ACTION: 30-Day Notice of Information Collection under Review: Form G- 845 and Form G-845 Supplement,...

  3. Mycelial forms of Pseudallescheria boydii present ectophosphatase activities.

    PubMed

    Kiffer-Moreira, Tina; Pinheiro, Ana Acacia S; Pinto, Márcia R; Esteves, Fabiano F; Souto-Padrón, Thais; Barreto-Bergter, Eliana; Meyer-Fernandes, José R

    2007-08-01

    Phosphatase activities were characterized in intact mycelial forms of Pseudallescheria boydii, which are able to hydrolyze the artificial substrate p-nitrophenylphosphate (p-NPP) to p-nitrophenol (p-NP) at a rate of 41.41+/-2.33 nmol p-NP per h per mg dry weight, linearly with increasing time and with increasing cell density. MgCl2, MnCl2 and ZnCl2 were able to increase the (p-NPP) hydrolysis while CdCl2 and CuCl2 inhibited it. The (p-NPP) hydrolysis was enhanced by increasing pH values (2.5-8.5) over an approximately 5-fold range. High sensitivity to specific inhibitors of alkaline and acid phosphatases suggests the presence of both acid and alkaline phosphatase activities on P. boydii mycelia surface. Cytochemical localization of the acid and alkaline phosphatase showed electron-dense cerium phosphate deposits on the cell wall, as visualized by electron microscopy. The product of p-NPP hydrolysis, inorganic phosphate (Pi), and different inhibitors for phosphatase activities inhibited p-NPP hydrolysis in a dose-dependent manner, but only the inhibition promoted by sodium orthovanadate and ammonium molybdate is irreversible. Intact mycelial forms of P. boydii are also able to hydrolyze phosphoaminoacids with different specificity.

  4. NUCLEAR ACTIVITY IS MORE PREVALENT IN STAR-FORMING GALAXIES

    SciTech Connect

    Rosario, D. J.; Lutz, D.; Berta, S.; Popesso, P.; Genzel, R.; Saintonge, A.; Tacconi, L.; Wuyts, S. E-mail: lutz@mpe.mpg.de E-mail: popesso@mpe.mpg.de E-mail: amelie@mpe.mpg.de E-mail: swuyts@mpe.mpg.de; and others

    2013-07-01

    We explore the question of whether low and moderate luminosity active galactic nuclei (AGNs) are preferentially found in galaxies that are undergoing a transition from active star formation (SF) to quiescence. This notion has been suggested by studies of the UV-optical colors of AGN hosts, which find them to be common among galaxies in the so-called Green Valley, a region of galaxy color space believed to be composed mostly of galaxies undergoing SF quenching. Combining the deepest current X-ray and Herschel/PACS far-infrared (FIR) observations of the two Chandra Deep Fields with redshifts, stellar masses, and rest-frame photometry derived from the extensive and uniform multi-wavelength data in these fields, we compare the rest-frame U - V color distributions and star formation rate distributions of AGNs and carefully constructed samples of inactive control galaxies. The UV-to-optical colors of AGNs are consistent with equally massive inactive galaxies at redshifts out to z {approx} 2, but we show that such colors are poor tracers of SF. While the FIR distributions of both star-forming AGNs and star-forming inactive galaxies are statistically similar, we show that AGNs are preferentially found in star-forming host galaxies, or, in other words, AGNs are less likely to be found in weakly star-forming or quenched galaxies. We postulate that, among X-ray-selected AGNs of low and moderate accretion luminosities, the supply of cold gas primarily determines the accretion rate distribution of the nuclear black holes.

  5. 75 FR 16492 - Agency Information Collection Activities: Form G-28, and Form G-28I, Revision of an Existing...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-01

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form G-28, and Form G- 28I, Revision of an Existing Information Collection; Comment Request ACTION: 60-Day Notice of Information Collection under Review: Form G- 28, Notice of Entry of Appearance as Attorney or...

  6. Optically active surfaces formed by ion implantation and thermal treatment

    SciTech Connect

    Gea, L.A.; Boatner, L.A.; Evans, H.M.; Zuhr, R.

    1996-08-01

    Embedded VO{sub 2} precipitates have been formed in single-crystal sapphire by the ion co-implantation of vanadium and oxygen and subsequent thermal annealing. The embedded VO{sub 2} particles have been shown to exhibit an optical switching behavior that is comparable to that of continuous thin films. In this work, the mechanisms of formation of these optically active particles are investigated. It is shown that precipitation of the vanadium dioxide phase is favored when the thermal treatment is performed on an ion-damaged but still crystalline (rather than amorphized) Al{sub 2}O{sub 3} substrate. The best optical switching behavior is observed in this case, and this behavior is apparently correlated with a more-favorable dispersion of VO{sub 2} small particles inside the matrix.

  7. Tectonic activity on Pluto after the Charon-forming impact

    NASA Astrophysics Data System (ADS)

    Barr, Amy C.; Collins, Geoffrey C.

    2015-01-01

    The Pluto-Charon system, likely formed from an impact, has reached the endpoint of its tidal evolution. During its evolution into the dual-synchronous state, the equilibrium tidal figures of Pluto and Charon would have also evolved as angular momentum was transferred from Pluto's spin to Charon's orbit. The rate of tidal evolution is controlled by Pluto's interior physical and thermal state. We examine three interior models for Pluto: an undifferentiated rock/ice mixture, differentiated with ice above rock, and differentiated with an ocean. For the undifferentiated case without an ocean, the Pluto-Charon binary does not evolve to its current state unless its internal temperature Ti > 200K , which would likely lead to strong tidal heating, melting, and differentiation. Without an ocean, Pluto's interior temperature must be higher than 240 K for Charon to evolve on a time scale less than the age of the Solar System. Further tidal heating would likely create an ocean. If New Horizons finds evidence of ancient tidally-driven tectonic activity on either body, the most likely explanation is that Pluto had an internal ocean during Charon's orbital evolution.

  8. Observations of actively forming lava tubes and associated structures, Hawaii.

    NASA Technical Reports Server (NTRS)

    Greeley, R.

    1971-01-01

    Fluid basalts were erupted in August, 1970, from a vent near Alae Crater and flowed southeast. Forming exclusively in pahoehoe basalt, tubes in general evolve from lava channels by crustal formation, although some tubes develop directly from the vent. The observation discussed shows that channel crusts and tube roofs form in several ways. Lava channels usually form along the axis of highest velocity within the flow and are often centered along older lava channels, stream beds, rifts, grabens, or fracture zones.

  9. The Essay: Theory and Pedagogy for an Active Form.

    ERIC Educational Resources Information Center

    Heilker, Paul

    Calling for a radical reexamination of the traditional foundation of composition instruction--the thesis/support form, this book argues that the essay, with its informality, conversational tone, meditative mood, and integration of form and content, is better suited to developmental, epistemological, ideological, and feminist rhetorical…

  10. Phosphorylated nuclear receptor CAR forms a homodimer to repress its constitutive activity for ligand activation.

    PubMed

    Shizu, Ryota; Osabe, Makoto; Perera, Lalith; Moore, Rick; Sueyoshi, Tatsuya; Negishi, Masahiko

    2017-03-06

    Nuclear receptor CAR (NR1I3) regulates hepatic drug and energy metabolism as well as cell fate. Its activation can be a critical factor in drug-induced toxicity and disease development such as diabetes and tumors. CAR inactivates its constitutive activity by phosphorylation at threonine 38. Utilizing receptor for protein kinase 1 (RACK1) as the regulatory subunit, protein phosphatase PP2A dephosphorylates threonine 38 to activate CAR. Here we have demonstrated that CAR undergoes its homodimer-monomer conversion to regulate this dephosphorylation. By co-expressing two differently-tagged CAR proteins in Huh-7 cells, mouse primary hepatocytes and mouse livers, co-immunoprecipitation and two-dimensional gel electrophoresis revealed that CAR can form a homodimer in a configuration in which the PP2A/RACK1 binding site is buried within its dimer interface. Epidermal growth factor (EGF) was found to stimulate CAR homo-dimerization, thus constraining CAR in its inactive form. The agonistic ligand CITCO binds directly to the CAR homodimer and dissociates phosphorylated CAR into its monomer, exposing the PP2A/RACK1 binding site for dephosphorylation. Phenobarbital, which is not a CAR ligand, binds the EGF receptor, reversing the EGF signal to monomerize CAR for its indirect activation. Thus, the homodimer-monomer conversion is the underlying molecular mechanism that regulates CAR activation, by placing phosphorylated threonine 38 as the common target for both direct and in direct activation of CAR.

  11. 76 FR 41279 - Agency Information Collection Activities; Form I-864, Form I-864A, Form I-864EZ, and Form I-864W...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-13

    ..., Affidavit of Support Under Section 213A of the Act; Form I-864W, Intending Immigrant's Affidavit of Support... by family-based and certain employment-based immigrants to have the petitioning relative execute...

  12. 75 FR 26782 - Agency Information Collection Activities: Form I-864, Form I-864A, Form I-864EZ, and Form I-864W...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-12

    ... Act; Form I-864W, Intending Immigrant's Affidavit of Support Exemption; OMB Control No. 1615-0075. The... immigrants to have the petitioning relative execute an Affidavit of Support on their behalf. (5) An...

  13. 75 FR 51093 - Agency Information Collection Activities: Form I-864, Form I-864A, Form I-864EZ, and Form I-864W...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-18

    ...-864 EZ, Affidavit of Support Under Section 213A of the Act; Form I-864W, Intending Immigrant's... immigrants to have the petitioning relative execute an Affidavit of Support on their behalf. (5) An...

  14. Canonical nucleosome organization at promoters forms during genome activation.

    PubMed

    Zhang, Yong; Vastenhouw, Nadine L; Feng, Jianxing; Fu, Kai; Wang, Chenfei; Ge, Ying; Pauli, Andrea; van Hummelen, Paul; Schier, Alexander F; Liu, X Shirley

    2014-02-01

    The organization of nucleosomes influences transcriptional activity by controlling accessibility of DNA binding proteins to the genome. Genome-wide nucleosome binding profiles have identified a canonical nucleosome organization at gene promoters, where arrays of well-positioned nucleosomes emanate from nucleosome-depleted regions. The mechanisms of formation and the function of canonical promoter nucleosome organization remain unclear. Here we analyze the genome-wide location of nucleosomes during zebrafish embryogenesis and show that well-positioned nucleosome arrays appear on thousands of promoters during the activation of the zygotic genome. The formation of canonical promoter nucleosome organization is independent of DNA sequence preference, transcriptional elongation, and robust RNA polymerase II (Pol II) binding. Instead, canonical promoter nucleosome organization correlates with the presence of histone H3 lysine 4 trimethylation (H3K4me3) and affects future transcriptional activation. These findings reveal that genome activation is central to the organization of nucleosome arrays during early embryogenesis.

  15. The pore-forming toxin proaerolysin is activated by furin.

    PubMed

    Abrami, L; Fivaz, M; Decroly, E; Seidah, N G; Jean, F; Thomas, G; Leppla, S H; Buckley, J T; van der Goot, F G

    1998-12-04

    Aerolysin is secreted as an inactive dimeric precursor by the bacterium Aeromonas hydrophila. Proteolytic cleavage within a mobile loop near the C terminus of the protoxin is required for oligomerization and channel formation. This loop contains the sequence KVRRAR432, which should be recognized by mammalian proprotein convertases such as furin, PACE4, and PC5/6A. Here we show that these three proteases cleave proaerolysin after Arg-432 in vitro, yielding active toxin. We also investigated the potential role of these enzymes in the in vivo activation of the protoxin. We found that Chinese hamster ovary cells were able to convert the protoxin to aerolysin in the absence of exogenous proteases and that activation did not require internalization of the toxin. The furin inhibitor alpha1-antitrypsin Portland reduced the rate of proaerolysin activation in vivo, and proaerolysin processing was even further reduced in furin-deficient FD11 Chinese hamster ovary cells. The cells were also less sensitive to proaerolysin than wild type cells; however, transient transfection of FD11 cells with the cDNA encoding furin conferred normal sensitivity to the protoxin. Together these findings argue that furin catalyzes the cell-surface activation of proaerolysin in vivo.

  16. Forming a Learning Culture to Promote Fracture Prevention Activities

    ERIC Educational Resources Information Center

    Hjalmarson, Helene V.; Strandmark, Margaretha

    2012-01-01

    Purpose: The purpose of this paper is to explore interprofessional experiences of incorporating fracture prevention activities in clinical practice inspired by an empowerment approach. Design/methodology/approach: Data collection consisted primarily of focus groups interviews, systematized and analyzed by the grounded theory method. The study took…

  17. 76 FR 31972 - Agency Information Collection Activities: Form I-508 and Form I-508F, Extension of a Currently...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-02

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-508 and... and Immigration Services (USCIS), will be submitting the following information collection request for... Immigration Services. (4) Affected public who will be asked or required to respond, as well as a...

  18. How Lipid Membranes Affect Pore Forming Toxin Activity.

    PubMed

    Rojko, Nejc; Anderluh, Gregor

    2015-12-15

    Pore forming toxins (PFTs) evolved to permeate the plasma membrane of target cells. This is achieved in a multistep mechanism that usually involves binding of soluble protein monomer to the lipid membrane, oligomerization at the plane of the membrane, and insertion of part of the polypeptide chain across the lipid membrane to form a conductive channel. Introduced pores allow uncontrolled transport of solutes across the membrane, inflicting damage to the target cell. PFTs are usually studied from the perspective of structure-function relationships, often neglecting the important role of the bulk membrane properties on the PFT mechanism of action. In this Account, we discuss how membrane lateral heterogeneity, thickness, and fluidity influence the pore forming process of PFTs. In general, lipid molecules are more accessible for binding in fluid membranes due to steric reasons. When PFT specifically binds ordered domains, it usually recognizes a specific lipid distribution pattern, like sphingomyelin (SM) clusters or SM/cholesterol complexes, and not individual lipid species. Lipid domains were also suggested to act as an additional concentration platform facilitating PFT oligomerization, but this is yet to be shown. The last stage in PFT action is the insertion of the transmembrane segment across the membranes to build the transmembrane pore walls. Conformational changes are a spontaneous process, and sufficient free energy has to be available for efficient membrane penetration. Therefore, fluid bilayers are permeabilized more readily in comparison to highly ordered and thicker liquid ordered lipid phase (Lo). Energetically more costly insertion into the Lo phase can be driven by the hydrophobic mismatch between the thinner liquid disordered phase (Ld) and large protein complexes, which are unable to tilt like single transmembrane segments. In the case of proteolipid pores, membrane properties can directly modulate pore size, stability, and even selectivity. Finally

  19. [The dynamics of forming an active defensive reflex in cats].

    PubMed

    Fokin, V F

    1975-01-01

    Active defensive reflexes were elaborated in cats with pain stimulations of the forepaw by means of an electrical pricking device with a target attached to it. The elaboration was carried out during action of a flickering light used for the convenience of the EEG analysis. Repeated pain stimulation led to elaboration of an aggressive attacking reaction, chiefly manifested in the paw striking the target. At the beginning of the elaboration, passive-defensive reactions were manifest, which did not completely disappear even after formation of a stable attacking reflex. Two types of active defensive reflexes were elaborated: A-type reflex which helped the animal to get rid of the pain stimulation at the very beginning; B-type reflex which prevented the pain stimulation. The difference beteween these two types is discussed.

  20. Active Curved Polymers Form Vortex Patterns on Membranes

    NASA Astrophysics Data System (ADS)

    Denk, Jonas; Huber, Lorenz; Reithmann, Emanuel; Frey, Erwin

    2016-04-01

    Recent in vitro experiments with FtsZ polymers show self-organization into different dynamic patterns, including structures reminiscent of the bacterial Z ring. We model FtsZ polymers as active particles moving along chiral, circular paths by Brownian dynamics simulations and a Boltzmann approach. Our two conceptually different methods point to a generic phase behavior. At intermediate particle densities, we find self-organization into vortex structures including closed rings. Moreover, we show that the dynamics at the onset of pattern formation is described by a generalized complex Ginzburg-Landau equation.

  1. 75 FR 51095 - Agency Information Collection Activities: Form N-336; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-18

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-336... Collection under Review; Form N- 336, Request for Hearing on a Decision in Naturalization Proceedings Under... the Form N-336. Should USCIS decide to revise Form N-336 we will advise the public when we publish...

  2. 78 FR 17220 - Agency Information Collection Activities: Application for Naturalization, Form N-400; Revision of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-20

    ... for Naturalization, Form N-400; Revision of a Currently Approved Collection ACTION: 30-day notice... questions to Form N-400. These additional questions will allow USCIS to make more informed decisions on the eligibility of respondents to the form. Form N-400 is the final information collection activity that...

  3. 75 FR 21013 - Agency Information Collection Activities: Form N-644; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-22

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-644... Collection Under Review; Form N- 644, Application for Posthumous Citizenship; OMB Control No. 1615-0059. The..., USCIS will be evaluating whether to revise the Form N-644. Should USCIS decide to revise Form N-644...

  4. 76 FR 21913 - Agency Information Collection Activities: Form N-644; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-19

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-644... Collection Under Review: Form N- 644, Application for Posthumous Citizenship; OMB Control No. 1615-0059. The..., USCIS will be evaluating whether to revise the Form N-644. Should USCIS decide to revise Form N-644...

  5. 76 FR 81517 - Agency Information Collection Activities: Form I-131, Revision of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-28

    ... the Form/Collection: Application for Travel Document. (3) Agency form number, if any, and the... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-131... Collection Under Review: Form I- 131, Application for Travel Document. The Department of Homeland Security,...

  6. 77 FR 15787 - Agency Information Collection Activities: Form I-131, Revision of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-16

    ... the Form/Collection: Application for Travel Document. (3) Agency Form Number, if any, and the... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-131... Collection Under Review: Form I- 131, Application for Travel Document. The Department of Homeland Security,...

  7. 75 FR 76021 - Agency Information Collection Activities: Passenger List/Crew List (CBP Form I-418)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-07

    ... SECURITY Customs and Border Protection Agency Information Collection Activities: Passenger List/Crew List... concerning the Passenger List/Crew List (CBP Form I-418). This request for comment is being made pursuant to...: Passenger List/Crew List. OMB Number: 1651-0103. Form Number: CBP Form I-418. Abstract: CBP Form I-418...

  8. 78 FR 26648 - Agency Information Collection Activities: Passenger List/Crew List (CBP Form I-418)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-07

    ... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Passenger List/Crew... concerning the Passenger List/Crew List (CBP Form I-418). This request for comment is being made pursuant to...: Passenger List/Crew List. OMB Number: 1651-0103. Form Number: CBP Form I-418. Abstract: CBP Form I-418...

  9. 77 FR 2561 - Agency Information Collection Activities: Passenger List/Crew List (CBP Form I-418)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-18

    ... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Passenger List/Crew... concerning the Passenger List/Crew List (CBP Form I-418). This request for comment is being made pursuant to...: Passenger List/Crew List. OMB Number: 1651-0103. Form Number: CBP Form I-418. Abstract: CBP Form I-418...

  10. Heat of Hydration of Low Activity Cementitious Waste Forms

    SciTech Connect

    Nasol, D.

    2015-07-23

    During the curing of secondary waste grout, the hydraulic materials in the dry mix react exothermally with the water in the secondary low-activity waste (LAW). The heat released, called the heat of hydration, can be measured using a TAM Air Isothermal Calorimeter. By holding temperature constant in the instrument, the heat of hydration during the curing process can be determined. This will provide information that can be used in the design of a waste solidification facility. At the Savannah River National Laboratory (SRNL), the heat of hydration and other physical properties are being collected on grout prepared using three simulants of liquid secondary waste generated at the Hanford Site. From this study it was found that both the simulant and dry mix each had an effect on the heat of hydration. It was also concluded that the higher the cement content in the dry materials mix, the greater the heat of hydration during the curing of grout.

  11. Surface active complexes formed between keratin polypeptides and ionic surfactants.

    PubMed

    Pan, Fang; Lu, Zhiming; Tucker, Ian; Hosking, Sarah; Petkov, Jordan; Lu, Jian R

    2016-12-15

    Keratins are a group of important proteins in skin and hair and as biomaterials they can provide desirable properties such as strength, biocompatibility, and moisture regaining and retaining. The aim of this work is to develop water-soluble keratin polypeptides from sheep wool and then explore how their surface adsorption behaves with and without surfactants. Successful preparation of keratin samples was demonstrated by identification of the key components from gel electrophoresis and the reproducible production of gram scale samples with and without SDS (sodium dodecylsulphate) during wool fibre dissolution. SDS micelles could reduce the formation of disulphide bonds between keratins during extraction, reducing inter-molecular crosslinking and improving keratin polypeptide solubility. However, Zeta potential measurements of the two polypeptide batches demonstrated almost identical pH dependent surface charge distributions with isoelectric points around pH 3.5, showing complete removal of SDS during purification by dialysis. In spite of different solubility from the two batches of keratin samples prepared, very similar adsorption and aggregation behavior was revealed from surface tension measurements and dynamic light scattering. Mixing of keratin polypeptides with SDS and C12TAB (dodecyltrimethylammonium bromide) led to the formation of keratin-surfactant complexes that were substantially more effective at reducing surface tension than the polypeptides alone, showing great promise in the delivery of keratin polypeptides via the surface active complexes. Neutron reflection measurements revealed the coexistence of surfactant and keratin polypeptides at the interface, thus providing the structural support to the observed surface tension changes associated with the formation of the surface active complexes.

  12. 78 FR 24187 - Commission Information Collection Activities (FERC Form 6-Q); Comment Request

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-24

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF ENERGY Federal Energy Regulatory Commission Commission Information Collection Activities (FERC Form 6-Q); Comment... Energy Regulatory Commission (Commission or FERC) is submitting the information collection FERC Form...

  13. Subunit interface mutants of rabbit muscle aldolase form active dimers.

    PubMed Central

    Beernink, P. T.; Tolan, D. R.

    1994-01-01

    We report the construction of subunit interface mutants of rabbit muscle aldolase A with altered quaternary structure. A mutation has been described that causes nonspherocytic hemolytic anemia and produces a thermolabile aldolase (Kishi H et al., 1987, Proc Natl Acad Sci USA 84:8623-8627). The disease arises from substitution of Gly for Asp-128, a residue at the subunit interface of human aldolase A. To elucidate the role of this residue in the highly homologous rabbit aldolase A, site-directed mutagenesis is used to replace Asp-128 with Gly, Ala, Asn, Gln, or Val. Rabbit aldolase D128G purified from Escherichia coli is found to be similar to human D128G by kinetic analysis, CD, and thermal inactivation assays. All of the mutant rabbit aldolases are similar to the wild-type rabbit enzyme in secondary structure and kinetic properties. In contrast, whereas the wild-type enzyme is a tetramer, chemical crosslinking and gel filtration indicate that a new dimeric species exists for the mutants. In sedimentation velocity experiments, the mutant enzymes as mixtures of dimer and tetramer at 4 degrees C. Sedimentation at 20 degrees C shows that the mutant enzymes are > 99.5% dimeric and, in the presence of substrate, that the dimeric species is active. Differential scanning calorimetry demonstrates that Tm values of the mutant enzymes are decreased by 12 degrees C compared to wild-type enzyme. The results indicate that Asp-128 is important for interface stability and suggest that 1 role of the quaternary structure of aldolase is to provide thermostability. PMID:7833800

  14. 75 FR 13776 - Agency Information Collection Activities: Form N-300; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-23

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-300... Collection Under Review; Form N- 300, Application to File Declaration of Intention; OMB Control No. 1615-0078... this 60-day period, USCIS will be evaluating whether to revise the Form N-300. Should USCIS decide...

  15. 76 FR 17144 - Agency Information Collection Activities: Form N-300; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-28

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-300... Collection Under Review; Form N- 300, Application to File Declaration of Intention; OMB Control No. 1615-0078... this 60 day period, USCIS will be evaluating whether to revise the Form N-300. Should USCIS decide...

  16. 76 FR 36560 - Agency Information Collection Activities; Form N-300; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-22

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities; Form N-300... Collection Under Review; Form N- 300, Application to File Declaration of Intention; OMB Control No. 1615-0078... Homeland Security sponsoring the collection: Form N-300; U.S. Citizenship and Immigration Services...

  17. 75 FR 13777 - Agency Information Collection Activities: Form I-824; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-23

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-824... Collection Under Review; Form I- 824; Application for Action on an Approved Application; OMB Control No. 1615... this 60-day period, USCIS will be evaluating whether to revise the Form I-824. Should USCIS decide...

  18. 77 FR 12072 - Agency Information Collection Activities: Form I-824; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-28

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-824... Collection Under Review; Form I- 824; Application for Action on an Approved Application. The Department..., 2012. During this 60 day period, USCIS will be evaluating whether to revise the Form I-824....

  19. 77 FR 27241 - Agency Information Collection Activities: Form I-824; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-09

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-824... Collection Under Review; Form I- 824; Application for Action on an Approved Application. The Department of... collection: Form I-824; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will...

  20. 75 FR 32801 - Agency Information Collection Activities: Form I-824; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-09

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-824... Collection Under Review; Form I- 824; Application for Action on an Approved Application; OMB Control No. 1615... Security sponsoring the collection: Form I-824; U.S. Citizenship and Immigration Services (USCIS)....

  1. 75 FR 70277 - Agency Information Collection Activities: Form N-336, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-17

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-336... Information Collection Under Review: Form N- 336, Request for Hearing on a Decision in Naturalization... collection: Form N-336; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will...

  2. 75 FR 78264 - Agency Information Collection Activities: Form N-336, Revision to an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-15

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-336... collection under review: Form N- 336, Request for Hearing on a Decision in Naturalization Proceedings Under... Register at 75 FR 51095 announcing the extension of the Form N-336. The 60-day notice announced that...

  3. 77 FR 18255 - Agency Information Collection Activities: Form N-565; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-27

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-565... collection under review; Form N- 565, Application for Replacement Naturalization/Citizenship Document; OMB... until May 29, 2012. During this 60 day period, USCIS will be evaluating whether to revise the Form...

  4. 75 FR 5098 - Agency Information Collection Activities: Form N-565, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-01

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-565... information collection under review: Form N- 565, Application for Replacement Naturalization/Citizenship... Homeland Security sponsoring the collection: Form N-565; U.S. Citizenship and Immigration Services...

  5. 75 FR 51096 - Agency Information Collection Activities: Form N-400; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-18

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-400... Collection Under Review; Form N- 400, Application for Naturalization; OMB Control No. 1615-0052. The... 60 day period, USCIS will be evaluating whether to revise the Form N-400. Should USCIS decide...

  6. 76 FR 69276 - Agency Information Collection Activities: Form N-336, Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-08

    ... SECURITY Citizenship and Immigration Services Agency Information Collection Activities: Form N-336... Information Collection Under Review: Form N- 336, Request for Hearing on a Decision in Naturalization...: Form N-336. U.S. Citizenship and Immigration Services. (4) Affected public who will be asked...

  7. 75 FR 43535 - Agency Information Collection Activities: Form N-644, Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-26

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-644... information collection under review: Form N- 644, Application for Posthumous Citizenship; OMB Control No. 1615... would be evaluating whether to revise the Form ] N-644. On July 15, 2010, USCIS published a...

  8. 76 FR 53144 - Agency Information Collection Activities: Form N-336; Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-25

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-336... Information Collection Under Review: Form N- 336, Request for Hearing on a Decision in Naturalization...: Form N-336; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will be asked...

  9. 75 FR 18871 - Agency Information Collection Activities: Form N-600K, Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-13

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-600K... Information Collection under Review: Form N- 600K, Application for Citizenship and Issuance of Certificate...: Form N-600K, U.S. Citizenship and Immigration Services. (4) Affected public who will be asked...

  10. 75 FR 30050 - Agency Information Collection Activities: Form N-648, Revision of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-28

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-648... Information Collection Under Review: Form N-648, Medical Certification for Disability Exceptions. OMB Control... applicable component of the Department of Homeland Security sponsoring the collection: Form N-648....

  11. 76 FR 78674 - Agency Information Collection Activities: Form N-470, Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-19

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-470... Information Collection Under Review: Form N- 470, Application To Preserve Residence for Naturalization; OMB..., and the applicable component of the Department of Homeland Security sponsoring the collection: Form...

  12. 75 FR 51096 - Agency Information Collection Activities: Form N-470; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-18

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-470... Collection Under Review; Form N- 470, Application To Preserve Residence for Naturalization; OMB Control No... until October 18, 2010. During this 60-day period, USCIS will be evaluating whether to revise the Form...

  13. 76 FR 63320 - Agency Information Collection Activities: Form N-470, Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-12

    ... Agency Information Collection Activities: Form N-470, Revision of a Currently Approved Information Collection; Comment Request ACTION: 60-Day Notice of Information Collection Under Review: Form N- 470... revision of Form N-470 instead of extending the current edition. USCIS invites members of the public...

  14. 76 FR 11807 - Agency Information Collection Activities: Form N-565, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-03

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-565... Information Collection Under Review: Form N- 565, Application for Replacement Naturalization/Citizenship..., and the applicable component of the Department of Homeland Security sponsoring the collection: Form...

  15. 76 FR 39415 - Agency Information Collection Activities: Form N-644, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-06

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-644... Information Collection Under Review: Form N- 644, Application for Posthumous Citizenship; OMB Control No. 1615... Homeland Security sponsoring the collection: Form N-644; U.S. Citizenship and Immigration Services...

  16. 75 FR 51094 - Agency Information Collection Activities: Form N-600; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-18

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-600... Collection under Review; Form N- 600, Application for Certificate of Citizenship; OMB Control No. 1615- 0057..., 2010. During this 60 day period, USCIS will be evaluating whether to revise the Form N-600....

  17. 77 FR 128 - Agency Information Collection Activities: Form N-600, Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-03

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-600... Information Collection Under Review: Form N- 600, Application for Certificate of Citizenship. The Department..., and the applicable component of the Department of Homeland Security sponsoring the collection: Form...

  18. 75 FR 71451 - Agency Information Collection Activities: Form N-470, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-23

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-470... Information Collection Under Review: Form N- 470, Application to Preserve Residence for Naturalization; OMB... collection: Form N-470; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will...

  19. 75 FR 70277 - Agency Information Collection Activities: Form N-400, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-17

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-400... Information Collection Under Review: Form N- 400, Application for Naturalization; OMB Control No. 1615-0052... applicable component of the Department of Homeland Security sponsoring the collection: Form N-400;...

  20. 76 FR 69275 - Agency Information Collection Activities: Form N-400, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-08

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-400... Information Collection Under Review: Form N- 400, Application for Naturalization; OMB Control No. 1615-0052... the Department of Homeland Security sponsoring the collection: Form N-400. U.S. Citizenship...

  1. 76 FR 58029 - Agency Information Collection Activities: Form N-600K, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-19

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-600K... Information Collection Under Review: Form N- 600K; Application for Citizenship and Issuance of Certificate... collection: Form N-600K; U.S. Citizenship and Immigration Services. (4) Affected public who will be asked...

  2. 75 FR 70278 - Agency Information Collection Activities: Form N-600, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-17

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-600... Information Collection Under Review: Form N- 600, Application for Certificate of Citizenship; OMB Control No..., and the applicable component of the Department of Homeland Security sponsoring the collection: Form...

  3. 76 FR 38197 - Agency Information Collection Activities; Form N-600K, Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-29

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities; Form N-600K... information collection under review: form N- 600K, application for citizenship and issuance of certificate... sponsoring the collection: Form N-600K, U.S. Citizenship and Immigration Services. (4) Affected public...

  4. 75 FR 5099 - Agency Information Collection Activities: Form N-648, Revision of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-01

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-648... information collection under review: Form N- 648, Medical Certification for Disability Exceptions. OMB Control... component of the Department of Homeland Security sponsoring the collection: Form N-648. U.S. Citizenship...

  5. 77 FR 24507 - Agency Information Collection Activities: Form N-25, Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-24

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-25... Information Collection Under Review: Form N- 25, Request for Verification of Naturalization. The Department of... component of the Department of Homeland Security sponsoring the collection: Form N-25. U.S. Citizenship...

  6. 75 FR 32800 - Agency Information Collection Activities: Form N-300; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-09

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-300... Collection Under Review; Form N- 300, Application To File Declaration of Intention; OMB Control No. 1615-0078... collection: Form N-300; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will...

  7. 75 FR 80835 - Agency Information Collection Activities: Form N-565; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-23

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-565... Collection Under Review; Form N- 565, Application for Replacement Naturalization/Citizenship Document; OMB... N-565. Should USCIS decide to revise Form N-565 we will advise the public when we publish the...

  8. 76 FR 59710 - Agency Information Collection Activities: Form N-600; Revision of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-27

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-600... Collection Under Review; Form N- 600, Application for Certificate of Citizenship; OMB Control No. 1615- 0057... the Department of Homeland Security sponsoring the collection: Form N-600; U.S. Citizenship...

  9. 76 FR 43336 - Agency Information Collection Activities: Form AR-11, Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-20

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form AR-11... Collection under Review: Form AR- 11, Alien's Change of Address Card; OMB Control No. 1615-0007. The... applicable component of the Department of Homeland Security sponsoring this collection: Form AR-11....

  10. 77 FR 12071 - Agency Information Collection Activities: Form G-28, Revision of a Currently Approved Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-28

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form G-28... Information Collection Under Review: Form G- 28, Notice of Entry of Appearance as Attorney or Accredited... other technological collection techniques, or other forms of information technology, e.g.,...

  11. 76 FR 28444 - Agency Information Collection Activities: Form G-884, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-17

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form G-884... information collection under review: Form G- 884, Request for the Return of Original Documents; OMB Control No... July 18, 2011. During this 60-day period, USCIS will be evaluating whether to revise the Form...

  12. 75 FR 47822 - Agency Information Collection Activities: Form G-639, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-09

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form G-639... Information Collection Under Review: Form G- 639, Freedom of Information/Privacy Act Request; OMB Control No... other forms of information technology, e.g., permitting electronic submission of responses. Overview...

  13. 76 FR 63322 - Agency Information Collection Activities: Form G-28, Revision of an Approved Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-12

    ... Information Collection Activities: Form G-28, Revision of an Approved Information Collection; Comment Request ACTION: 60-Day Notice of Information Collection Under Review: Form G- 28, Notice of Entry of Appearance..., mechanical, or other technological collection techniques, or other forms of information technology,...

  14. 76 FR 24908 - Agency Information Collection Activities: Form G-639; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-03

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form G-639... Collection Under Review; Form G- 639, Freedom of Information/Privacy Act Request; OMB Control No. 1615- 0102.... During this 60 day period, USCIS will be evaluating whether to revise the Form G-639. Should USCIS...

  15. 75 FR 23785 - Agency Information Collection Activities: Form G-639; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-04

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form G-639... Collection Under Review; Form G- 639, Freedom of Information/Privacy Act Request; OMB Control No. 1615- 0102... this 60 day period, USCIS will be evaluating whether to revise the Form G-639. Should USCIS decide...

  16. Composition and topology of activity cliff clusters formed by bioactive compounds.

    PubMed

    Stumpfe, Dagmar; Dimova, Dilyana; Bajorath, Jürgen

    2014-02-24

    The assessment of activity cliffs has thus far mostly focused on compound pairs, although the majority of activity cliffs are not formed in isolation but in a coordinated manner involving multiple active compounds and cliffs. However, the composition of coordinated activity cliff configurations and their topologies are unknown. Therefore, we have identified all activity cliff configurations formed by currently available bioactive compounds and analyzed them in network representations where activity cliff configurations occur as clusters. The composition, topology, frequency of occurrence, and target distribution of activity cliff clusters have been determined. A limited number of large cliff clusters with unique topologies were identified that were centers of activity cliff formation. These clusters originated from a small number of target sets. However, most clusters were of small to moderate size. Three basic topologies were sufficient to describe recurrent activity cliff cluster motifs/topologies. For example, frequently occurring clusters with star topology determined the scale-free character of the global activity cliff network and represented a characteristic activity cliff configuration. Large clusters with complex topology were often found to contain different combinations of basic topologies. Our study provides a first view of activity cliff configurations formed by currently available bioactive compounds and of the recurrent topologies of activity cliff clusters. Activity cliff clusters of defined topology can be selected, and from compounds forming the clusters, SAR information can be obtained. The SAR information of activity cliff clusters sharing a/one specific activity and topology can be compared.

  17. Structural Differences between Active Forms of Plasminogen Activator Inhibitor Type 1 Revealed by Conformationally Sensitive Ligands*

    PubMed Central

    Li, Shih-Hon; Gorlatova, Natalia V.; Lawrence, Daniel A.; Schwartz, Bradford S.

    2008-01-01

    Plasminogen activator inhibitor type 1 (PAI-1) is a serine protease inhibitor (serpin) in which the reactive center loop (RCL) spontaneously inserts into a central β-sheet, β-sheet A, resulting in inactive inhibitor. Available x-ray crystallographic studies of PAI-1 in an active conformation relied on the use of stabilizing mutations. Recently it has become evident that these structural models do not adequately explain the behavior of wild-type PAI-1 (wtPAI-1) in solution. To probe the structure of native wtPAI-1, we used three conformationally sensitive ligands: the physiologic cofactor, vitronectin; a monoclonal antibody, 33B8, that binds preferentially to RCL-inserted forms of PAI-1; and RCL-mimicking peptides that insert into β-sheet A. From patterns of interaction with wtPAI-1 and the stable mutant, 14-1B, we propose a model of the native conformation of wtPAI-1 in which the bottom of the central sheet is closed, whereas the top of the β-sheet A is open to allow partial insertion of the RCL. Because the incorporation of RCL-mimicking peptides into wtPAI-1 is accelerated by vitronectin, we further propose that vitronectin alters the conformation of the RCL to allow increased accessibility to β-sheet A, yielding a structural hypothesis that is contradictory to the current structural model of PAI-1 in solution and its interaction with vitronectin. PMID:18436534

  18. PARP promoter-mediated activation of a VSG expression site promoter in insect form Trypanosoma brucei.

    PubMed

    Urményi, T P; Van der Ploeg, L H

    1995-03-25

    In trypanosomes the rRNA, PARP and VSG gene promoters mediate alpha-amanitin-resistant transcription of protein coding genes, presumably by RNA polymerase (pol) I. We compared the activity of PARP and VSG promoters integrated at one of the alleles of the largest subunit of pol II genes in insect form trypanosomes. Even though both promoters are roughly equally active in transient transformation assays in insect form trypanosomes, only the PARP promoter functioned effectively when integrated at the pol II largest subunit or other loci. Promoter activity in transient transformation assays is therefore not necessarily predictive of transcriptional activity once integrated into the trypanosome genome. The integrated fully active PARP promoter could upregulate in cis an otherwise poorly active integrated VSG promoter. The PARP promoter nucleotide sequence elements responsible for VSG promoter activation coincided with most of the important PARP promoter elements mapped previously by linker scanning mutagenesis, indicating that it is not a single unique promoter element that was responsible for VSG promoter activation. The data suggest that PARP promoter-mediated activation of the VSG promoter does not result from complementation of the VSG promoter with a single insect form-specific transcription factor whose binding site is missing from the VSG promoter and present in the PARP promoter. We favor a model in which chromatin structure at the locus is altered by the PARP promoter, allowing VSG promoter activation in insect form trypanosomes. We discuss the significance of these observations for the control of VSG promoters in insect form trypanosomes.

  19. 77 FR 27539 - Reports, Forms, and Recordkeeping Requirements; Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-10

    ... instrumented motorcycles. The National Highway Traffic Safety Administration (NHTSA) will be conducting on-road... National Highway Traffic Safety Administration Reports, Forms, and Recordkeeping Requirements; Agency Information Collection Activity Under OMB Review AGENCY: National Highway Traffic Safety Administration...

  20. 76 FR 66127 - Reports, Forms and Recordkeeping Requirements; Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-25

    ... Maritime Administration Reports, Forms and Recordkeeping Requirements; Agency Information Collection Activity Under OMB Review AGENCY: Maritime Administration, DOT. ACTION: Notice and request for comments... November 25, 2011. FOR FURTHER INFORMATION CONTACT: Dennis Brennan, Maritime Administration, 1200...

  1. 76 FR 24561 - Reports, Forms and Recordkeeping Requirements; Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-02

    ... Maritime Administration Reports, Forms and Recordkeeping Requirements; Agency Information Collection Activity Under OMB Review AGENCY: Maritime Administration, DOT. ACTION: Notice and request for comments... Administration (MARAD). Title: MARAD's Marine Transportation Economic Impact Model Data Needs. OMB Control...

  2. 77 FR 27473 - Agency Information Collection Activities: Form I-924; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-10

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-924... Department of Homeland Security (DHS), U.S. Citizenship and Immigration Services (USCIS) will be submitted... sponsoring the collection: Form I-924; U.S. Citizenship and Immigration Services. (4) Affected public...

  3. 76 FR 9805 - Agency Information Collection Activities: Form G-845 and Supplement; Revision of a Currently...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-22

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form G-845 and.... Citizenship and Immigration Services (USCIS) will be submitting the following information collection request... collection: Form G-845 and Supplement. U.S. Citizenship and Immigration Services. (4) Affected public...

  4. 76 FR 70747 - Agency Information Collection Activities: Form I-90; Revision of a Currently Approved Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-15

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-90... Department of Homeland Security, U.S. Citizenship and Immigration Services (USCIS) will be submitting the...: Form I-90; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will be asked...

  5. 76 FR 52961 - Agency Information Collection Activities: Form N-300; Revision of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-24

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-300.... The Department Homeland Security, U.S. Citizenship and Immigration Services (USCIS) will be submitting... collection: Form N-300; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will...

  6. 76 FR 31971 - Agency Information Collection Activities: Form I-212; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-02

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-212.... Citizenship and Immigration Services (USCIS) will be submitting the following information collection request... Homeland Security sponsoring the collection: Form I-212; U.S. Citizenship and Immigration Services...

  7. 76 FR 72209 - Agency Information Collection Activities: Form N-300; Revision of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-22

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-300... Homeland Security, U.S. Citizenship and Immigration Services (USCIS) will be submitting the following... Homeland Security sponsoring the collection: Form N-300; U.S. Citizenship and Immigration Services...

  8. 77 FR 16047 - Agency Information Collection Activities: Form I-589; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-19

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-589.... 1615-0067. The Department Homeland Security, U.S. Citizenship and Immigration Services (USCIS) will be...: Form I-589; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will be asked...

  9. 77 FR 34398 - Agency Information Collection Activities: Form N-565, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-11

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-565... Immigration Services (USCIS) will be submitting the following information collection request to the Office of... the collection: Form N-565; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public...

  10. 75 FR 41216 - Agency Information Collection Activities: Form N-644, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-15

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-644... Information Collection Under Review: Form N- 644, Application for Posthumous Citizenship; OMB Control No. 1615... N-644; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will be asked...

  11. 77 FR 65702 - Agency Information Collection Activities: Refugee/Asylee Relative Petition, Form Number I-730...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-30

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Refugee/Asylee... time required to complete this benefit request. (2) Title of the Form/Collection: Refugee/Asylee... abstract: Primary: Individuals or households. Form I- 730 will be used by an asylee or refugee to file...

  12. 78 FR 5477 - Agency Information Collection Activities: InfoPass System, No Form Number; Extension, Without...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-25

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: InfoPass... Collection. (2) Title of the Form/Collection: InfoPass System. (3) Agency form number, if any, and the.... The InfoPass system allows an applicant or petitioner to schedule an interview appointment with...

  13. 78 FR 52824 - Proposed Information Collection (Bowel and Bladder Care Billing Form) Activity: Comment Request

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-26

    ... AFFAIRS Proposed Information Collection (Bowel and Bladder Care Billing Form) Activity: Comment Request... evaluate the Bowel and Bladder Care Billing Form used by caregivers of eligible Veterans to document time spent providing services related specifically to bowel and bladder care. DATES: Written comments...

  14. 76 FR 53929 - Agency Information Collection Activities: Form G-639, Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-30

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form G-639... Information Collection Under Review: Form G- 639, Freedom of Information/Privacy Act Request. * * * * * The... information technology, e.g., permitting electronic submission of responses. Overview of this...

  15. 76 FR 48874 - Agency Information Collection Activities: Form G-884, Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-09

    ... SECURITY Citizenship and Immigration Services Agency Information Collection Activities: Form G-884... Collection Under Review: Form G- 884, Request for the Return of Original Document(s). The Department of..., e.g., permitting electronic submission of responses. Overview of This Information Collection...

  16. 77 FR 66958 - Agency Information Collection Activities; Comment Request; Form for Maintenance of Effort Waiver...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-08

    ... Agency Information Collection Activities; Comment Request; Form for Maintenance of Effort Waiver Requests... notice will be considered public records. Title of Collection: Form for Maintenance of Effort Waiver... (SEA) finds that either the combined fiscal effort per student or the aggregate expenditures of the...

  17. 76 FR 9810 - Agency Information Collection Activities: Comment Request for the Ferrous Metals Surveys (17 Forms)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-22

    ... Geological Survey Agency Information Collection Activities: Comment Request for the Ferrous Metals Surveys... to supply the USGS with domestic consumption data of 13 ores, concentrates, metals, and ferroalloys... OMB Control Number: 1028-0068. Form Number: Various (17 forms). Title: Ferrous Metals Surveys. Type...

  18. Effect of chemical form of selenium on tissue glutathione peroxidase activity in developing rats

    NASA Technical Reports Server (NTRS)

    Lane, Helen W.; Strength, Ralph; Johnson, Janet; White, Marguerite T.

    1991-01-01

    The hypothesis that the stage of development of rats may affect the availability of various forms of selenium for the activity of glutathione peroxidase (GSHPx) in the rat was experimentally investigated. One experiment evaluated the availability of selenium as selenite or selenomethionine for GSPHx activity during three developmental states in rats: fetus and 7-day old and 14-day old nursing pups. In all tissues studied, GSHPx activity was highest in the 14-day-old pups whose dams were in the selenomethionine group. Rat pups given intraperitoneal selenite had higher liver and kidney GSHPx activity than pups given the same amount of selenium as intraperitoneal selenomethionine. In a second experiment, all dams were fed the same basal diet and pups were weaned to diets containing one of two levels of selenium and one of three forms of selenium (selenite, selenomethionine, or selenocystine). The results also supported the hypothesis these dietary forms of selenium are differentially available for GSHPx activity.

  19. Differential Expression of Extracellular Lipase and Protease Activities of Mycelial and Yeast Forms in Malassezia furfur.

    PubMed

    Juntachai, Weerapong; Kajiwara, Susumu

    2015-10-01

    Malassezia furfur is a dimorphic yeast that is part of the human skin microflora. This fungus is a pathogen of a certain skin diseases, such as pityriasis versicolor, and in rare cases causes systemic infection in neonates. However, the role of dimorphism in the pathogenicity remains unclear. A modified induction medium (IM) was successfully able to induce mycelial growth of M. furfur under both solid and liquid condition. Filamentous elements with branching hyphae were observed when cultured in the IM. Furthermore, addition of bovine fetus serum into the liquid IM did not promote hyphal formation; on the contrary, it retrograded hyphae to the yeast form. Plate-washing assay showed that M. furfur hyphae did not possess the ability of invasive growth. Secretory proteins from both yeast and hyphal forms were isolated, and lipase and protease activities were analyzed. Intriguingly, the hyphal form showed higher activities than those of the yeast form, particularly the protease activity.

  20. Effects of Three Forms of Reading-Based Output Activity on L2 Vocabulary Learning

    ERIC Educational Resources Information Center

    Rassaei, Ehsan

    2017-01-01

    The current study investigated the effects of three forms of output activity on EFL learners' recognition and recall of second language (L2) vocabulary. To this end, three groups of learners of English as a foreign language (EFL) were instructed to employ the following three output activities after reading two narrative texts: (1) summarizing the…

  1. Comparing Two Forms of Concept Map Critique Activities to Facilitate Knowledge Integration Processes in Evolution Education

    ERIC Educational Resources Information Center

    Schwendimann, Beat A.; Linn, Marcia C.

    2016-01-01

    Concept map activities often lack a subsequent revision step that facilitates knowledge integration. This study compares two collaborative critique activities using a Knowledge Integration Map (KIM), a form of concept map. Four classes of high school biology students (n?=?81) using an online inquiry-based learning unit on evolution were assigned…

  2. Education Technologies in Addressing the Problem of Forming the Socially Active Individual

    ERIC Educational Resources Information Center

    Popova, Irina N.

    2016-01-01

    The article is devoted to the analysis of technological support of the educational process in solving the problem of forming the socially active individual. The authors studied the value of the category "social activity" and analyzed educational technologies that have an impact on its formation. The obtained results gave the possibility…

  3. Differential cellulolytic activity of native-form and C-terminal tagged-form cellulase derived from coptotermes formosanus and expressed in E. coli

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The endogenous cellulase gene (CfEG3a) of Coptotermes formosanus, an economically important pest termite, was cloned and overexpressed in both native form (nCfEG) and C-terminal His-tagged form (tCfEG) in E.coli. Both forms of recombinant cellulases showed hydrolytic activity on cellulosic substrate...

  4. Solubilization of the O2(-)-forming activity responsible for the respiratory burst in human neutrophils.

    PubMed

    Gabig, T G; Kipnes, R S; Babior, B M

    1978-10-10

    On exposure to suitable activating agents, neutrophils sharply alter their oxygen metabolism, showing large increases in oxygen uptake, O2 and H2O2 production, and glucose consumption via the hexose monophosphate shunt. These metabolic alterations, which together are designated the "respiratory burst," are due to the activation of a system which catalyzes the reaction: 2O2 + NADPH leads to 2O2(-) + NADP. This O2(-)-forming system is found in a particulate fraction isolated from neutrophils which had been activated with opsonized zymosan. When these particles were treated with detergent under suitable conditions, the O2(-)-forming activity was released in a form which passed through a membrane filter capable of retaining species of Mr greater than 3000,000. Soluble O2(-)-forming activity was obtained from normal activated neutrophils, but not from normal resting neutrophils or from activated neutrophils obtained from patients with chronic granulomatous disease, an inherited condition in which the respiratory burst is defective. O2(-)production by the soluble system required a reduced pyridine nucleotide as electron donor, and showed a quadratic dependence on the concentration of the solubilized preparation.

  5. Expression, purification and characterization of inactive and active forms of ERK2 from insect expression system.

    PubMed

    Yan, Kelly; Merritt, Hanne; Crawford, Kenneth; Pardee, Gwynn; Cheng, Jan Marie; Widger, Stephania; Hekmat-Nejad, Mohammad; Zaror, Isabel; Sim, Janet

    2015-06-01

    Extracellular signal-regulated kinase 2 (ERK2) is a serine/threonine protein kinase involved in many cellular programs, such as cell proliferation, differentiation, motility and programed cell-death. It is therefore considered an important target in the treatment of cancer. In an effort to support biochemical screening and small molecule drug discovery, we established a robust system to generate both inactive and active forms of ERK2 using insect expression system. We report here, for the first time, that inactive ERK2 can be expressed and purified with 100% homogeneity in the unphosphorylated form using insect system. This resulted in a significant 20-fold yield improvement compared to that previously reported using bacterial expression system. We also report a newly developed system to generate active ERK2 in insect cells through in vivo co-expression with a constitutively active MEK1 (S218D S222D). Isolated active ERK2 was confirmed to be doubly phosphorylated at the correct sites, T185 and Y187, in the activation loop of ERK2. Both ERK2 forms, inactive and active, were well characterized by biochemical activity assay for their kinase function. Inactive and active ERK2 were the two key reagents that enabled successful high through-put biochemical assay screen and structural drug discovery studies.

  6. [Antirestriction activity of T7 Ocr protein in monomeric and dimeric forms].

    PubMed

    Zavil'gelskiĭ, G B; Kotova, V Iu

    2014-01-01

    The Ocr protein, encoded by 0.3 (ocr) gene of bacteriophage T7, belongs to the family of antirestriction proteins that specifically inhibit the type I restriction-modification systems. Native Ocr forms homodimer (Ocr)2 both in solution and in the crystalline state. The Ocr protein belongs to the family of mimicry proteins. F53D A57E and E53R V77D mutant proteins were obtained, which form monomers. It was shown that the values of the dissociation constants Kd for Ocr, Ocr F53D A57E and Ocr F53RV77D proteins with EcoKI enzyme differ in 1000 times: Kd (Ocr) = 10(-10) M, Kd (Ocr F53D A57E and Ocr F53R V77D) = 10(-7) M. Antimodification activity of the Ocr monomeric forms is significantly reduced. We have shown, that Ocr dimeric form has fundamental importance for high inhibitory activity.

  7. Activated Transport in the Separate Layers that Form the νT=1 Exciton Condensate

    NASA Astrophysics Data System (ADS)

    Wiersma, R. D.; Lok, J. G.; Kraus, S.; Dietsche, W.; von Klitzing, K.; Schuh, D.; Bichler, M.; Tranitz, H.-P.; Wegscheider, W.

    2004-12-01

    We observe the total filling factor νT=1 quantum Hall state in a bilayer two-dimensional electron system with virtually no tunneling. We find thermally activated transport in the balanced system with a monotonic increase of the activation energy with decreasing d/ℓB below 1.65. In the imbalanced system we find activated transport in each of the layers separately, yet the activation energies show a striking asymmetry around the balance point, implying a different excitation spectrum for the separate layers forming the condensed state.

  8. Optical imaging of MMP-12 active form in inflammation and aneurysm

    PubMed Central

    Razavian, Mahmoud; Bordenave, Thomas; Georgiadis, Dimitris; Beau, Fabrice; Zhang, Jiasheng; Golestani, Reza; Toczek, Jakub; Jung, Jae-Joon; Ye, Yunpeng; Kim, Hye-Yeong; Han, Jinah; Dive, Vincent; Devel, Laurent; Sadeghi, Mehran M.

    2016-01-01

    Matrix metalloproteinase (MMP)-12 plays a key role in the development of aneurysm. Like other members of MMP family, MMP-12 is produced as a proenzyme, mainly by macrophages, and undergoes proteolytic activation to generate an active form. Accordingly, molecular imaging of the MMP-12 active form can inform of the pathogenic process in aneurysm. Here, we developed a novel family of fluorescent probes based on a selective MMP-12 inhibitor, RXP470.1 to target the active form of MMP-12. These probes were stable in complex media and retained the high affinity and selectivity of RXP470.1 for MMP-12. Amongst these, probe 3 containing a zwitterionic fluorophore, ZW800-1, combined a favorable affinity profile toward MMP-12 and faster blood clearance. In vivo binding of probe 3 was observed in murine models of sterile inflammation and carotid aneurysm. Binding specificity was demonstrated using a non-binding homolog. Co-immunostaining localized MMP-12 probe binding to MMP-12 positive areas and F4/80 positive macrophages in aneurysm. In conclusion, the active form of MMP-12 can be detected by optical imaging using RXP470.1-based probes. This is a valuable adjunct for pathophysiology research, drug development, and potentially clinical applications. PMID:27917892

  9. Membrane-associated forms of peptidylglycine alpha-amidating monooxygenase activity in rat pituitary. Tissue specificity.

    PubMed

    May, V; Cullen, E I; Braas, K M; Eipper, B A

    1988-06-05

    Membrane-associated peptidylglycine alpha-amidating monooxygenase (PAM) activity was investigated in rat anterior and neurointermediate pituitary tissues and in pituitary AtT-20/D-16v and GH3 cell lines. A substantial fraction of total pituitary PAM activity was found to be membrane-associated. Triton X-100, N-octyl-beta-D-glucopyranoside, and Zwittergent were effective in solubilizing PAM activity from crude pituitary membranes. The distribution of enzyme activity between soluble and membrane-associated forms was tissue-specific. In the anterior pituitary lobe and pituitary cell lines, 40-60% of total PAM activity was membrane-associated while only 10% of the alpha-amidating activity in the neurointermediate lobe was membrane-associated. Soluble and membrane-associated forms of PAM shared nearly identical characteristics with respect to copper and ascorbate requirements, pH optima, and Km values. Upon subcellular fractionation of anterior and neurointermediate pituitary lobe homogenates on Percoll gradients, 12-18% of total PAM activity was found in the rough endoplasmic reticulum/Golgi fractions and 42-60% was localized to secretory granule fractions. For both tissues, membrane-associated PAM activity was enriched in the rough endoplasmic reticulum/Golgi pool, whereas most of the secretory granule-associated enzyme activity was soluble.

  10. Synthesis of catalytically active form III ribulose 1,5-bisphosphate carboxylase/oxygenase in archaea.

    PubMed

    Finn, Michael W; Tabita, F Robert

    2003-05-01

    Ribulose 1,5 bisphosphate carboxylase/oxygenase (RubisCO) catalyzes the biological reduction and assimilation of carbon dioxide gas to organic carbon; it is the key enzyme responsible for the bulk of organic matter found on earth. Until recently it was believed that there are only two forms of RubisCO, form I and form II. However, the recent completion of several genome-sequencing projects uncovered open reading frames resembling RubisCO in the third domain of life, the archaea. Previous work and homology comparisons suggest that these enzymes represent a third form of RubisCO, form III. While earlier work indicated that two structurally distinct recombinant archaeal RubisCO proteins catalyzed bona fide RubisCO reactions, it was not established that the rbcL genes of anaerobic archaea can be transcribed and translated to an active enzyme in the native organisms. In this report, it is shown not only that Methanococcus jannaschii, Archaeoglobus fulgidus, Methanosarcina acetivorans, and Methanosarcina barkeri possess open reading frames with the residues required for catalysis but also that the RubisCO protein from these archaea accumulates in an active form under normal growth conditions. In addition, the form III RubisCO gene (rbcL) from M. acetivorans was shown to complement RubisCO deletion strains of Rhodobacter capsulatus and Rhodobacter sphaeroides under both photoheterotrophic and photoautotrophic growth conditions. These studies thus indicate for the first time that archaeal form III RubisCO functions in a physiologically significant fashion to fix CO(2). Furthermore, recombinant M. jannaschii, M. acetivorans, and A. fulgidus RubisCO possess unique properties with respect to quaternary structure, temperature optima, and activity in the presence of molecular oxygen compared to the previously described Thermococcus kodakaraensis and halophile proteins.

  11. Hepatic overexpression of a constitutively active form of liver glycogen synthase improves glucose homeostasis.

    PubMed

    Ros, Susana; Zafra, Delia; Valles-Ortega, Jordi; García-Rocha, Mar; Forrow, Stephen; Domínguez, Jorge; Calbó, Joaquim; Guinovart, Joan J

    2010-11-26

    In this study, we tested the efficacy of increasing liver glycogen synthase to improve blood glucose homeostasis. The overexpression of wild-type liver glycogen synthase in rats had no effect on blood glucose homeostasis in either the fed or the fasted state. In contrast, the expression of a constitutively active mutant form of the enzyme caused a significant lowering of blood glucose in the former but not the latter state. Moreover, it markedly enhanced the clearance of blood glucose when fasted rats were challenged with a glucose load. Hepatic glycogen stores in rats overexpressing the activated mutant form of liver glycogen synthase were enhanced in the fed state and in response to an oral glucose load but showed a net decline during fasting. In order to test whether these effects were maintained during long term activation of liver glycogen synthase, we generated liver-specific transgenic mice expressing the constitutively active LGS form. These mice also showed an enhanced capacity to store glycogen in the fed state and an improved glucose tolerance when challenged with a glucose load. Thus, we conclude that the activation of liver glycogen synthase improves glucose tolerance in the fed state without compromising glycogenolysis in the postabsorptive state. On the basis of these findings, we propose that the activation of liver glycogen synthase may provide a potential strategy for improvement of glucose tolerance in the postprandial state.

  12. Functional Specificity of the Visual Word Form Area: General Activation for Words and Symbols but Specific Network Activation for Words

    ERIC Educational Resources Information Center

    Reinke, Karen; Fernandes, Myra; Schwindt, Graeme; O'Craven, Kathleen; Grady, Cheryl L.

    2008-01-01

    The functional specificity of the brain region known as the Visual Word Form Area (VWFA) was examined using fMRI. We explored whether this area serves a general role in processing symbolic stimuli, rather than being selective for the processing of words. Brain activity was measured during a visual 1-back task to English words, meaningful symbols…

  13. Project Activities as a Form of English Language Teaching Based on the Interdisciplinary Approach to Form Intercultural Communicative Competence

    ERIC Educational Resources Information Center

    Redchenko, Nadezhda N.

    2016-01-01

    The authors of this article suggest a thesis about the purpose of teaching a foreign language--it is student's communicative activities, i.e. learning a foreign language in practice. The teacher's task is to encourage activities of every student and to create situations to develop their creative activities in a learning process. New information…

  14. 76 FR 53144 - Agency Information Collection Activities: Form I-508 and Form I-508F, Extension of a Currently...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-25

    ..., Exemptions and Immunities. The Department of Homeland Security, U.S. Citizenship and Immigration Services..., Privileges, Exemptions and Immunities. (3) Agency form number, if any, and the applicable component of...

  15. 77 FR 27474 - Agency Information Collection Activities: Form I-687; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-10

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-687...-687. The Department Homeland Security (DHS), U.S. Citizenship and Immigration Services (USCIS) has...-687; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will be asked...

  16. 77 FR 34052 - Agency Information Collection Activities: Form I-102; Revision of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-08

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-102... Document. The Department of Homeland Security, U.S. Citizenship and Immigration Services (USCIS) will be... Immigration Services (USCIS). (4) Affected public who will be asked or required to respond, as well as a...

  17. 77 FR 2560 - Agency Information Collection Activities: Form I-90, Revision of a Currently Approved Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-18

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-90... Department of Homeland Security, U.S. Citizenship and Immigration Services (USCIS) will be submitting the.... Citizenship and Immigration Services. (4) Affected public who will be asked or required to respond, as well...

  18. 77 FR 64119 - Agency Information Collection Activities: Refugee/Asylee Adjusting Status, Form Number I-643...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-18

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Refugee/Asylee...: 30-Day Notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration... form when applying for adjustment of status, with the U.S. Citizenship and Immigration Services...

  19. 78 FR 26647 - Agency Information Collection Activities: Application To Replace Permanent Resident Card, Form I...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-07

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Application To... Notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration Services... Security sponsoring the collection: Form I-90, U.S. Citizenship and Immigration Services. (4)...

  20. 76 FR 70747 - Agency Information Collection Activities: Form I-526, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-15

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-526... Homeland Security, U.S. Citizenship and Immigration Services (USCIS) will be submitting the following... Immigration Services. (4) Affected public who will be asked or required to respond, as well as a...

  1. 77 FR 50710 - Agency Information Collection Activities: Employment Eligibility Verification, Form I-9, OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-22

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Employment... Verification, Form I-9. The Department of Homeland Security (DHS), U.S. Citizenship and Immigration Services... Immigration Services. (4) Affected public who will be asked or required to respond, as well as a...

  2. 77 FR 34053 - Agency Information Collection Activities: Form I-590, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-08

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-590... of Homeland Security, U.S. Citizenship and Immigration Services (USCIS) will be submitting the...-590; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will be asked...

  3. 75 FR 65022 - Agency Information Collection Activities: Form I-751, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-21

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-751.... 1615- 0038. The Department of Homeland Security, U.S. Citizenship and Immigration Services (USCIS) will...-751; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public who will be asked...

  4. 75 FR 65022 - Agency Information Collection Activities: Form I-698, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-21

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-698... Resident; OMB Control No. 1615-0035. The Department of Homeland Security, U.S. Citizenship and Immigration... Immigration Services (USCIS). (4) Affected public who will be asked or required to respond, as well as a...

  5. 77 FR 27474 - Agency Information Collection Activities: Form N-648, Revision of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-10

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form N-648... Department of Homeland Security (DHS), U.S. Citizenship and Immigration Services (USCIS) will be submitting.... Citizenship and Immigration Services (USCIS). (4) Affected public who will be asked or required to respond,...

  6. 75 FR 57480 - Agency Information Collection Activities: Customs Declaration (Form 6059B)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-21

    ... SECURITY U.S. Customs And Border Protection Agency Information Collection Activities: Customs Declaration (Form 6059B) AGENCY: U.S. Customs and Border Protection (CBP), Department of Homeland Security. ACTION... the Customs Declaration. This request for comment is being made pursuant to the Paperwork...

  7. 75 FR 71452 - Agency Information Collection Activities: Customs Declaration (Form 6059B)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-23

    ... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Customs Declaration (Form 6059B) AGENCY: U.S. Customs and Border Protection, Department of Homeland Security. ACTION: 30-Day.... Customs and Border Protection (CBP) of the Department of Homeland Security will be submitting...

  8. 77 FR 65898 - Agency Information Collection Activities: InfoPass System, No Form Number; Extension, Without...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-31

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: InfoPass...: Extension, Without Change, of a Currently Approved Collection. (2) Title of the Form/Collection: InfoPass... as a brief abstract: Primary: Individuals or households. The InfoPass system allows an applicant...

  9. 75 FR 5066 - Commission Information Collection Activities (FERC Form 60,1

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-01

    ... information (including balance sheet, assets, liabilities, billing and charges for associated and non... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF ENERGY Federal Energy Regulatory Commission Commission Information Collection Activities (FERC Form 60,\\1\\ FERC-61,...

  10. 75 FR 3245 - Agency Information Collection Activities: Aircraft/Vessel Report (Form I-92)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-20

    ... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Aircraft/Vessel Report (Form I-92) AGENCY: U.S. Customs and Border Protection, Department of Homeland Security. ACTION.... SUMMARY: U.S. Customs and Border Protection (CBP) of the Department of Homeland Security has submitted...

  11. 75 FR 5099 - Agency Information Collection Activities: Visa Waiver Program Carrier Agreement (Form I-775)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-01

    ... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Visa Waiver Program Carrier Agreement (Form I-775) AGENCY: U.S. Customs and Border Protection, Department of Homeland Security.... SUMMARY: U.S. Customs and Border Protection (CBP) of the Department of Homeland Security has submitted...

  12. 76 FR 9806 - Agency Information Collection Activities: Passenger List/Crew List (CBP Form I-418)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-22

    ... Management and Budget. Comments should be addressed to the OMB Desk Officer for Customs and Border Protection... SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Passenger List/Crew List (CBP Form I-418) AGENCY: U.S. Customs and Border Protection, Department of Homeland...

  13. 75 FR 1069 - Agency Information Collection Activities: Passenger List/Crew List (Form I-418)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-08

    ...: 2010-107] DEPARTMENT OF HOMELAND SECURITY U.S. Customs and Border Protection Agency Information Collection Activities: Passenger List/Crew List (Form I-418) AGENCY: U.S. Customs and Border Protection... information collection: 1651-0103. SUMMARY: U.S. Customs and Border Protection (CBP) of the Department...

  14. PROCESS FOR THE PRODUCTION OF AN ACTIVATED FORM OF UO$sub 2$

    DOEpatents

    Polissar, M.J.

    1957-09-24

    A process for producing a highly active form of UO/sub 2/ characterized both by rapid oxidation in air and by rapid chlorination with CCl/sub 4/ vapor at an elevated temperature is reported. In accordance with the process, commercial UO/sub 2/, is subjected to a series of oxidation-reduction operations to produce a form of UC/sub 2/ of enhanced reactivity. By treatimg commercial UO/sub 2/ at a temperature between 335 and 485 deg C with methane, then briefly with an oxygen containing gas and followimg this by a second treatment with a methane containing gas, the original relatively stable charge of UO/sub 2/ will be transformed into an active form of UO/sub 2/.

  15. 76 FR 25364 - Agency Information Collection Activities: Form I-864, Form I-864A, Form I-864EZ, and From I-864W...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-04

    ..., Affidavit of Support Under Section 213A of the Act; Form I-864W, Intending Immigrant's Affidavit of Support...-based immigrants to have the petitioning relative execute an Affidavit of Support on their behalf....

  16. Identification of dimethyl disulfide-forming bacteria isolated from activated sludge.

    PubMed Central

    Tomita, B; Inoue, H; Chaya, K; Nakamura, A; Hamamura, N; Ueno, K; Watanabe, K; Ose, Y

    1987-01-01

    Twenty-four strains with high dimethyl disulfide (DMDS)-forming ability were isolated from activated sludge and identified to the genus level. These bacteria were classified into four groups (A, B, C, and D) by the API ZYM System (API System S.A., Montalieu, France). Group A (three strains) was identified as genus Lactobacillus by the API 20B System, by the method of Cowan and Steel, and by production of lactic acid as confirmed by gas-liquid chromatography. Group B (eight strains) was identified as genus Corynebacterium by API 20B and the Cowan and Steel method. Group C (one strain) was suggested to belong to genus Corynebacterium by the API 20B System. Group D (12 strains) was identified as genus Pseudomonas or Alcaligenes by the API 20B System, as genus Alcaligenes by the Cowan and Steel method, and as Achromobacter group Vd by the API 20NE System. However, on the basis of guanine-plus-cytosine contents in DNA and form of flagella, these strains were identified as genus Pseudomonas. Formation of DMDS from DL-methionine and S-methyl-L-cysteine was tested. DMDS-forming bacteria isolated from activated sludge formed DMDS from both precursors. In genus Pseudomonas, P. aeruginosa could not form DMDS from either precursor, but P. acidovorans, P. alcaligenes, P. pseudoalcaligenes, and P. testosteroni formed DMDS. In genus Alcaligenes, A. denitrificans subsp. xylosoxydans, A. denitrificans subsp. denitrificans, A. faecalis, and A. odorans formed DMDS from both precursors. Achromobacter group Vd formed DMDS from S-methyl-L-cysteine, but could not from DL-methionine. PMID:3662505

  17. Binary Toxin Subunits of Lysinibacillus sphaericus Are Monomeric and Form Heterodimers after In Vitro Activation

    PubMed Central

    Choong, Yeu Khai; Torres, Jaume; Boonserm, Panadda

    2016-01-01

    The binary toxin from Lysinibacillus sphaericus has been successfully used for controlling mosquito-transmitted diseases. An activation step shortens both subunits BinA and BinB before their interaction with membranes and internalization in midgut cells, but the precise role of this activation step is unknown. Herein, we show conclusively using three orthogonal biophysical techniques that protoxin subunits form only monomers in aqueous solution. However, in vitro activated toxins readily form heterodimers. This oligomeric state did not change after incubation of these heterodimers with detergent. These results are consistent with the evidence that maximal toxicity in mosquito larvae is achieved when the two subunits, BinA and BinB, are in a 1:1 molar ratio, and directly link proteolytic activation to heterodimerization. Formation of a heterodimer must thus be necessary for subsequent steps, e.g., interaction with membranes, or with a suitable receptor in susceptible mosquito species. Lastly, despite existing similarities between BinB C-terminal domain with domains 3 and 4 of pore-forming aerolysin, no aerolysin-like SDS-resistant heptameric oligomers were observed when the activated Bin subunits were incubated in the presence of detergents or lipidic membranes. PMID:27341696

  18. Creating a Physical Activity Self-Report Form for Youth Using Rasch Methodology.

    PubMed

    DiStefano, Christine; Pate, Russell; McIver, Kerry; Dowda, Marsha; Beets, Michael; Murrie, Dale

    2016-01-01

    Measurement of youth's physical activity levels is recommended to ensure that children are meeting recommended activity guidelines. This article describes the creation of an instrument to measure youth's levels of physical activity, where a strong test validation perspective (Benson, 1998) was followed to create the scale. The development process involved a mixed-method (qualitative followed by quantitative) framework. First, focus groups were conducted, where results informed item creation. Next, three alternative forms were created with different response formats to measure childrens' frequency of participation in various physical activities and intensity of participation. Lastly, a sample of over 500 middle school children was obtained, where three different response scales were investigated. The optimal scale considered measurement of physical activity using a three-point Likert frequency; intensity of activity participation did not strongly contribute to the measurement of children's activity levels. The final version form is thought to be acceptable for use with children in surveillance and large-group studies, as well as in smaller sample applications.

  19. Creating a Physical Activity Self-Report Form for Youth Using Rasch Methodology

    PubMed Central

    DiStefano, Christine; Pate, Russell; McIver, Kerry; Dowda, Marsha; Beets, Michael; Murrie, Dale

    2016-01-01

    Measurement of youth’s physical activity levels is recommended to ensure that children are meeting recommended activity guidelines. This article describes the creation of an instrument to measure youth’s levels of physical activity, where a strong test validation perspective (Benson, 1998) was followed to create the scale. The development process involved a mixed-method (qualitative followed by quantitative) framework. First, focus groups were conducted where results informed item creation. Next, three alternative forms were created with different response formats to measure children’s’ frequency of participation in various physical activities and intensity of participation. Lastly, a sample of over 500 middle school children was obtained where three different response scales were investigated. The optimal scale considered measurement of physical activity using a three-point Likert frequency; intensity of activity participation did not strongly contribute to the measurement of children’s activity levels. The final version form is thought to be acceptable for use with children in surveillance and large-group studies, as well as in smaller sample applications. PMID:28009580

  20. Cytochrome P450-mediated activation of the fragrance compound geraniol forms potent contact allergens

    SciTech Connect

    Hagvall, Lina; Baron, Jens Malte; Boerje, Anna; Weidolf, Lars; Merk, Hans; Karlberg, Ann-Therese

    2008-12-01

    Contact sensitization is caused by low molecular weight compounds which penetrate the skin and bind to protein. In many cases, these compounds are activated to reactive species, either by autoxidation on exposure to air or by metabolic activation in the skin. Geraniol, a widely used fragrance chemical, is considered to be a weak allergen, although its chemical structure does not indicate it to be a contact sensitizer. We have shown that geraniol autoxidizes and forms allergenic oxidation products. In the literature, it is suggested but not shown that geraniol could be metabolically activated to geranial. Previously, a skin-like CYP cocktail consisting of cutaneous CYP isoenzymes, was developed as a model system to study cutaneous metabolism. In the present study, we used this system to investigate CYP-mediated activation of geraniol. In incubations with the skin-like CYP cocktail, geranial, neral, 2,3-epoxygeraniol, 6,7-epoxygeraniol and 6,7-epoxygeranial were identified. Geranial was the main metabolite formed followed by 6,7-epoxygeraniol. The allergenic activities of the identified metabolites were determined in the murine local lymph node assay (LLNA). Geranial, neral and 6,7-epoxygeraniol were shown to be moderate sensitizers, and 6,7-epoxygeranial a strong sensitizer. Of the isoenzymes studied, CYP2B6, CYP1A1 and CYP3A5 showed high activities. It is likely that CYP1A1 and CYP3A5 are mainly responsible for the metabolic activation of geraniol in the skin, as they are expressed constitutively at significantly higher levels than CYP2B6. Thus, geraniol is activated through both autoxidation and metabolism. The allergens geranial and neral are formed via both oxidation mechanisms, thereby playing a large role in the sensitization to geraniol.

  1. The proteolytic system of pineapple stems revisited: Purification and characterization of multiple catalytically active forms.

    PubMed

    Matagne, André; Bolle, Laetitia; El Mahyaoui, Rachida; Baeyens-Volant, Danielle; Azarkan, Mohamed

    2017-02-23

    Crude pineapple proteases extract (aka stem bromelain; EC 3.4.22.4) is an important proteolytic mixture that contains enzymes belonging to the cysteine proteases of the papain family. Numerous studies have been reported aiming at the fractionation and characterization of the many molecular species present in the extract, but more efforts are still required to obtain sufficient quantities of the various purified protease forms for detailed physicochemical, enzymatic and structural characterization. In this work, we describe an efficient strategy towards the purification of at least eight enzymatic forms. Thus, following rapid fractionation on a SP-Sepharose FF column, two sub-populations with proteolytic activity were obtained: the unbound (termed acidic) and bound (termed basic) bromelain fractions. Following reversible modification with monomethoxypolyethylene glycol (mPEG), both fractions were further separated on Q-Sepharose FF and SP-Sepharose FF, respectively. This procedure yielded highly purified molecular species, all titrating ca. 1 mol of thiol group per mole of enzyme, with distinct biochemical properties. N-terminal sequencing allowed identifying at least eight forms with proteolytic activity. The basic fraction contained previously identified species, i.e. basic bromelain forms 1 and 2, ananain forms 1 and 2, and comosain (MEROPS identifier: C01.027). Furthermore, a new proteolytic species, showing similarities with basic bomelain forms 1 and 2, was discovered and termed bromelain form 3. The two remaining species were found in the acidic bromelain fraction and were arbitrarily named acidic bromelain forms 1 and 2. Both, acidic bromelain forms 1, 2 and basic bromelain forms 1, 2 and 3 are glycosylated, while ananain forms 1 and 2, and comosain are not. The eight protease forms display different amidase activities against the various substrates tested, namely small synthetic chromogenic compounds (DL-BAPNA and Boc-Ala-Ala-Gly-pNA), fluorogenic compounds

  2. Integration of active pharmaceutical ingredient solid form selection and particle engineering into drug product design.

    PubMed

    Ticehurst, Martyn David; Marziano, Ivan

    2015-06-01

    This review seeks to offer a broad perspective that encompasses an understanding of the drug product attributes affected by active pharmaceutical ingredient (API) physical properties, their link to solid form selection and the role of particle engineering. While the crucial role of active pharmaceutical ingredient (API) solid form selection is universally acknowledged in the pharmaceutical industry, the value of increasing effort to understanding the link between solid form, API physical properties and drug product formulation and manufacture is now also being recognised. A truly holistic strategy for drug product development should focus on connecting solid form selection, particle engineering and formulation design to both exploit opportunities to access simpler manufacturing operations and prevent failures. Modelling and predictive tools that assist in establishing these links early in product development are discussed. In addition, the potential for differences between the ingoing API physical properties and those in the final product caused by drug product processing is considered. The focus of this review is on oral solid dosage forms and dry powder inhaler products for lung delivery.

  3. Structure of the active form of human origin recognition complex and its ATPase motor module

    PubMed Central

    Tocilj, Ante; On, Kin Fan; Yuan, Zuanning; Sun, Jingchuan; Elkayam, Elad; Li, Huilin; Stillman, Bruce; Joshua-Tor, Leemor

    2017-01-01

    Binding of the Origin Recognition Complex (ORC) to origins of replication marks the first step in the initiation of replication of the genome in all eukaryotic cells. Here, we report the structure of the active form of human ORC determined by X-ray crystallography and cryo-electron microscopy. The complex is composed of an ORC1/4/5 motor module lobe in an organization reminiscent of the DNA polymerase clamp loader complexes. A second lobe contains the ORC2/3 subunits. The complex is organized as a double-layered shallow corkscrew, with the AAA+ and AAA+-like domains forming one layer, and the winged-helix domains (WHDs) forming a top layer. CDC6 fits easily between ORC1 and ORC2, completing the ring and the DNA-binding channel, forming an additional ATP hydrolysis site. Analysis of the ATPase activity of the complex provides a basis for understanding ORC activity as well as molecular defects observed in Meier-Gorlin Syndrome mutations. DOI: http://dx.doi.org/10.7554/eLife.20818.001 PMID:28112645

  4. 76 FR 46781 - Commission Information Collection Activities (FERC Form 1 and FERC Form 1F); Comment Request...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-03

    ..., and 3Q \\2\\. The FERC Annual/Quarterly Report Forms provide the Commission, as well as others, with an... financial and operating report submitted for electric rate regulation and financial audits. Major is defined as having (1) One million Megawatt hours or more; (2) 100 megawatt hours of annual sales for...

  5. Crystallization and preliminary analysis of active nitroalkane oxidase in three crystal forms

    PubMed Central

    Nagpal, Akanksha; Valley, Michael P.; Fitzpatrick, Paul F.; Orville, Allen M.

    2006-01-01

    Nitroalkane oxidase (NAO), a flavoprotein cloned and purified from Fusarium oxysporum, catalyzes the oxidation of neutral nitroalkanes to the corresponding aldehydes or ketones, with the production of H2O2 and nitrite. In this paper, the crystallization and preliminary X-ray data analysis of three crystal forms of active nitroalkane oxidase are described. The first crystal form belongs to a trigonal space group (either P3121 or P3221, with unit-cell parameters a = b = 103.8, c = 487.0 Å) and diffracts to at least 1.6 Å resolution. Several data sets were collected using 2θ and κ geometry in order to obtain a complete data set to 2.07 Å resolution. Solvent-content and Matthews coefficient analysis suggests that crystal form 1 contains two homotetramers per asymmetric unit. Crystal form 2 (P212121; a = 147.3, b = 153.5, c = 169.5 Å) and crystal form 3 (P31 or P32; a = b = 108.9, c = 342.5 Å) are obtained from slightly different conditions and also contain two homotetramers per asymmetric unit, but have different solvent contents. A three-wavelength MAD data set was collected from selenomethionine-enriched NAO (SeMet-NAO) in crystal form 3 and will be used for phasing. PMID:15272176

  6. Amino acid sequence of homologous rat atrial peptides: natriuretic activity of native and synthetic forms.

    PubMed Central

    Seidah, N G; Lazure, C; Chrétien, M; Thibault, G; Garcia, R; Cantin, M; Genest, J; Nutt, R F; Brady, S F; Lyle, T A

    1984-01-01

    A substance called atrial natriuretic factor (ANF), localized in secretory granules of atrial cardiocytes, was isolated as four homologous natriuretic peptides from homogenates of rat atria. The complete sequence of the longest form showed that it is composed of 33 amino acids. The three other shorter forms (2-33, 3-33, and 8-33) represent amino-terminally truncated versions of the 33 amino acid parent molecule as shown by analysis of sequence, amino acid composition, or both. The proposed primary structure agrees entirely with the amino acid composition and reveals no significant sequence homology with any known protein or segment of protein. The short form ANF-(8-33) was synthesized by a multi-fragment condensation approach and the synthetic product was shown to exhibit specific activity comparable to that of the natural ANF-(3-33). PMID:6232612

  7. One-step purification of a functional, constitutively activated form of visual arrestin.

    PubMed

    Huang, Li; Mao, Xiang; Abdulaev, Najmoutin G; Ngo, Tony; Liu, Wei; Ridge, Kevin D

    2012-03-01

    Desensitization of agonist-activated G protein-coupled receptors (GPCRs) requires phosphorylation followed by the binding of arrestin, a ~48 kDa soluble protein. While crystal structures for the inactive, 'basal' state of various arrestins are available, the conformation of 'activated' arrestin adopted upon interaction with activated GPCRs remains unknown. As a first step towards applying high-resolution structural methods to study arrestin conformation and dynamics, we have utilized the subtilisin prodomain/Profinity eXact™ fusion-tag system for the high-level bacterial expression and one-step purification of wild-type visual arrestin (arrestin 1) as well as a mutant form (R175E) of the protein that binds to non-phosphorylated, light-activated rhodopsin (Rho∗). The results show that both prodomain/Profinity eXact™ fusion-tagged wild-type and R175E arrestins can be expressed to levels approaching 2-3 mg/l in Luria-Bertani media, and that the processed, tag-free mature forms can be purified to near homogeneity using a Bio-Scale™ Mini Profinity eXact™ cartridge on the Profinia™ purification system. Functional analysis of R175E arrestin generated using this approach shows that it binds to non-phosphorylated rhodopsin in a light-dependent manner. These findings should facilitate the structure determination of this 'constitutively activated' state of arrestin 1 as well as the monitoring of conformational changes upon interaction with Rho∗.

  8. Protease-Activated Pore-Forming Peptides for the Treatment and Imaging of Prostate Cancer

    PubMed Central

    LeBeau, Aaron M.; Denmeade, Samuel R.

    2015-01-01

    A common hallmark of cancers with highly aggressive phenotypes is increased proteolysis in the tumor and the surrounding microenvironment. Prostate cancer has a number of proteases uniquely associated with it that may play various important roles in disease progression. In this report, we utilize the peritumoral proteolytic activity of prostate cancer to activate engineered peptide constructs for the treatment and noninvasive imaging of prostate cancer. Using a modular "propeptide" approach, a cationic diastereomeric pore-forming peptide domain was linked to an inactivating acidic peptide domain. The inactivating acidic peptide domain was engineered to be a cleavable substrate for the secreted serine protease prostate-specific antigen (PSA) or the transmembrane metalloprotease prostate-specific membrane antigen (PSMA). The propeptides were then evaluated in a direct comparison study. Both the PSA and PSMA activated propeptides were found to be cytotoxic to prostate cancer cells in vitro. In vivo, however, treatment of LNCaP and CWR22Rv1 xenografts with the PSMA propeptide resulted in a pronounced cytostatic effect when compared with xenografts treated with the PSA propeptide or the cationic diastereomeric peptide alone. The PSMA activated propeptide also proved to be an effective optical imaging probe in vivo when labeled with a near-infrared fluorophore. These data suggest that protease-activated pore-forming peptides could potentially be used for both imaging and treating prostate cancer. PMID:25537662

  9. Antimicrobial and Antioxidant Activity of Chitosan/Hydroxypropyl Methylcellulose Film-Forming Hydrosols Hydrolyzed by Cellulase

    PubMed Central

    Zimoch-Korzycka, Anna; Bobak, Łukasz; Jarmoluk, Andrzej

    2016-01-01

    The aim of this study was to evaluate the impact of cellulase (C) on the biological activity of chitosan/hydroxypropyl methylcellulose (CH/HPMC) film-forming hydrosols. The hydrolytic activity of cellulase in two concentrations (0.05% and 0.1%) was verified by determination of the progress of polysaccharide hydrolysis, based on viscosity measurement and reducing sugar-ends assay. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging effect, the ferric reducing antioxidant power (FRAP), and microbial reduction of Pseudomonas fluorescens, Yersinia enterocolitica, Bacillus cereus, and Staphylococcus aureus were studied. During the first 3 h of reaction, relative reducing sugar concentration increased progressively, and viscosity decreased rapidly. With increasing amount of enzyme from 0.05% to 0.1%, the reducing sugar concentration increased, and the viscosity decreased significantly. The scavenging effect of film-forming solutions was improved from 7.6% at time 0 and without enzyme to 52.1% for 0.1% cellulase after 20 h of reaction. A significant effect of cellulase addition and reaction time on antioxidant power of the tested film-forming solutions was also reported. Film-forming hydrosols with cellulase exhibited a bacteriostatic effect on all tested bacteria, causing a total reduction. PMID:27608008

  10. Validity and Reliability of International Physical Activity Questionnaire-Short Form in Chinese Youth

    ERIC Educational Resources Information Center

    Wang, Chao; Chen, Peijie; Zhuang, Jie

    2013-01-01

    Purpose: The psychometric profiles of the widely used International Physical Activity Questionnaire-Short Form (IPAQ-SF) in Chinese youth have not been reported. The purpose of this study was to examine the validity and reliability of the IPAQ-SF using a sample of Chinese youth. Method: One thousand and twenty-one youth (M[subscript age] = 14.26 ±…

  11. A Natural Chimeric Pseudomonas Bacteriocin with Novel Pore-Forming Activity Parasitizes the Ferrichrome Transporter.

    PubMed

    Ghequire, Maarten G K; Kemland, Lieselore; Anoz-Carbonell, Ernesto; Buchanan, Susan K; De Mot, René

    2017-02-21

    Modular bacteriocins represent a major group of secreted protein toxins with a narrow spectrum of activity, involved in interference competition between Gram-negative bacteria. These antibacterial proteins include a domain for binding to the target cell and a toxin module at the carboxy terminus. Self-inhibition of producers is provided by coexpression of linked immunity genes that transiently inhibit the toxin's activity through formation of bacteriocin-immunity complexes or by insertion in the inner membrane, depending on the type of toxin module. We demonstrate strain-specific inhibitory activity for PmnH, a Pseudomonas bacteriocin with an unprecedented dual-toxin architecture, hosting both a colicin M domain, potentially interfering with peptidoglycan synthesis, and a novel colicin N-type domain, a pore-forming module distinct from the colicin Ia-type domain in Pseudomonas aeruginosa pyocin S5. A downstream-linked gene product confers PmnH immunity upon susceptible strains. This protein, ImnH, has a transmembrane topology similar to that of Pseudomonas colicin M-like and pore-forming immunity proteins, although homology with either of these is essentially absent. The enhanced killing activity of PmnH under iron-limited growth conditions reflects parasitism of the ferrichrome-type transporter for entry into target cells, a strategy shown here to be used as well by monodomain colicin M-like bacteriocins from pseudomonads. The integration of a second type of toxin module in a bacteriocin gene could offer a competitive advantage against bacteria displaying immunity against only one of both toxic activities.IMPORTANCE In their continuous struggle for ecological space, bacteria face a huge load of contenders, including phylogenetically related strains that compete for the same niche. One important group of secreted antibacterial proteins assisting in eliminating these rivals are modular bacteriocins of Gram-negative bacteria, comprising a domain for docking onto the

  12. Pore-forming activity of alpha-toxin is essential for clostridium septicum-mediated myonecrosis.

    PubMed

    Kennedy, Catherine L; Lyras, Dena; Cordner, Leanne M; Melton-Witt, Jody; Emmins, John J; Tweten, Rodney K; Rood, Julian I

    2009-03-01

    Clostridium septicum alpha-toxin is a beta-barrel pore-forming cytolysin that is functionally similar to aerolysin. Residues important in receptor binding, oligomerization, and pore formation have been identified; however, little is known about the activity of the toxin in an infection, although it is essential for disease. We have now shown that deletion of a small portion of the transmembrane domain, so that the toxin is no longer able to form pores, completely abrogates its ability to contribute to disease, as does replacement of the sole cysteine residue with leucine. However, although previous biochemical and cytotoxicity assays clearly indicated that mutations in residues important in oligomerization, binding, and prepore conversion greatly reduced activity or rendered the toxin inactive, once the mutated toxins were overexpressed by the natural host in the context of an infection it was found they were able to cause disease in a mouse model of myonecrosis. These results highlight the importance of testing the activity of virulence determinants in the normal host background and in an infectious disease context and provide unequivocal evidence that it is the ability of alpha-toxin to form a pore that confers its toxicity in vivo.

  13. Anthrax toxin: channel-forming activity of protective antigen in planar phospholipid bilayers.

    PubMed Central

    Blaustein, R O; Koehler, T M; Collier, R J; Finkelstein, A

    1989-01-01

    The three separate proteins that make up anthrax toxin--protective antigen (PA), edema factor (EF), and lethal factor (LF)--act in binary combinations to produce two distinct reactions in experimental animals: edema (PA + EF) and death (PA + LF). PA is believed to interact with a membrane receptor, and after proteolytic processing, to mediate endocytosis and subsequent translocation of EF or LF into the cytosol. PA can be separated, after mild trypsinolysis, into two fragments, PA65 (65 kDa) and PA20 (20 kDa). We demonstrate that trypsin-cleaved PA is capable of forming cation-selective channels in planar phospholipid bilayer membranes and that this activity is confined to the PA65 fragment; PA20, LF, and EF are devoid of channel-forming activity. These PA65 channels exhibit pH-dependent and voltage-dependent activity--a property reminiscent of the channels formed by the two-chain proteins diphtheria, tetanus, and botulinum toxins. Images PMID:2467303

  14. Nurse eggs form through an active process of apoptosis in the spionid Polydora cornuta (Annelida).

    PubMed

    Gibson, Glenys; Hart, Corban; Coulter, Claire; Xu, Haixin

    2012-07-01

    The production of nurse eggs is fundamental to poecilogony in some species of spionid annelids. In species such as Polydora cornuta, nurse-egg production varies among females and ingestion of nurse eggs varies among young, resulting in a form of poecilogony with divergent phenotypes for females (e.g., fecundity and per-offspring investment) as well as for larvae (e.g., trophic mode, size, and stage at hatching). We tested the hypothesis that nurse eggs of P. cornuta form through an active developmental process and specifically, through apoptosis. Results of a TUNEL assay indicate nuclear fragmentation occurs in a process that is characteristic of apoptosis. Cellular indicators of apoptosis in nurse eggs include activation of caspase-3, a positive Annexin V reaction indicating exposure of phosphatidylserine on the outer cell membrane, and invagination of the membrane to form yolk vesicles. These results indicate that formation of nurse eggs in this population of P. cornuta occurs through an active, adaptive process. Furthermore, while apoptosis also occurs in some cells of P. cornuta embryos, it was not detected until later in development. This suggests that nurse eggs originate through heterochrony in a developmental process (apoptosis) that is common to all young of P. cornuta.

  15. 77 FR 9259 - Agency Information Collection Activities: Forms G-1041 and G-1041A, Extension of a Currently...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-16

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Forms G-1041... Notice of Information Collection Under Review: Forms G- 1041 and G-1041A, Genealogy Index Search Request... evaluating whether to revise forms G-1041 and G-1041A. Should USCIS decide to revise forms G- 1041 and...

  16. Effects of pharmaceutical processing on pepsin activity during the formulation of solid dosage forms.

    PubMed

    Kristó, Katalin; Pintye-Hódi, Klára

    2013-02-01

    The main aim of this study was to investigate the effects of pharmaceutical technological methods on pepsin activity during the formulation of solid dosage forms. The circumstances of direct compression and wet granulation were modeled. During direct compression, the heat and the compression force must be taken into consideration. The effects of these parameters were investigated in three materials (pure pepsin, and 1:1 (w/w) pepsin-tartaric acid and 1:1 (w/w) pepsin-citric acid powder mixtures). It was concluded that direct compression is appropriate for the formulation of solid dosage forms containing pepsin through application without acids or with acids at low compression force. The effects of wet granulation were investigated with a factorial design for the same three materials. The factors were time, temperature and moisture content. There was no significant effect of the factors when acids were not applied. Temperature was a significant factor when acids were applied. The negative effect was significantly higher for citric acid than for tartaric acid. It was found that wet granulation can be utilized for the processing of pepsin into solid dosage forms under well-controlled circumstances. The application of citric acid is not recommended during the formulation of solid dosage forms through wet granulation. A mathematically based optimization may be necessary for preformulation studies of the preparation of dosage forms containing sensitive enzymes.

  17. Differential range and activity of various forms of the Hedgehog protein

    PubMed Central

    Dawber, Rebecca J; Hebbes, Stephen; Herpers, Bram; Docquier, France; van den Heuvel, Marcel

    2005-01-01

    Background The Hedgehog (Hh) family of secreted proteins act as extracellular messengers to control and coordinate growth and differentiation. The mechanism by which Hh protein travels across a field of cells, and results in a range of specific effects relating to the distance from the source, has been the subject of much debate. It has been suggested that the range and activity of the pathway can be linked to modifications of the Hh protein, specifically the addition of lipid groups at N- and C-terminal sites. Results Here we have addressed the potency of different forms of Hh protein by expressing these in Drosophila, where we are able to precisely establish pathway activity and range in naïve but responsive tissues. As expected, a construct that can produce all forms of Hh recapitulates endogenous signaling potencies. In comparison, expression of a form that lacks the cholesterol moiety (HhN) leads to an extended range, but the product is less effective at inducing maximal Hh responses. Expression of a point mutant that lacks the N-terminal palmitate binding site shows that the palmitoylation of Hh is absolutely required for activity in this system. Conclusion We conclude that the addition of the cholesterol moiety limits the range of the protein and is required for maximal activity, while addition of palmitate is required for all activity. These findings have implications for understanding how Hedgehog proteins move, and thus their potential at influencing distant sites, and concomitantly, how modifications of the signaling protein can affect the efficacy of the response in exposed cells. PMID:16197551

  18. Theoretical studies of chemical reactivity of metabolically activated forms of aromatic amines toward DNA.

    PubMed

    Shamovsky, Igor; Ripa, Lena; Blomberg, Niklas; Eriksson, Leif A; Hansen, Peter; Mee, Christine; Tyrchan, Christian; O'Donovan, Mike; Sjö, Peter

    2012-10-15

    The metabolism of aromatic and heteroaromatic amines (ArNH₂) results in nitrenium ions (ArNH⁺) that modify nucleobases of DNA, primarily deoxyguanosine (dG), by forming dG-C8 adducts. The activated amine nitrogen in ArNH⁺ reacts with the C8 of dG, which gives rise to mutations in DNA. For the most mutagenic ArNH₂, including the majority of known genotoxic carcinogens, the stability of ArNH⁺ is of intermediate magnitude. To understand the origin of this observation as well as the specificity of reactions of ArNH⁺ with guanines in DNA, we investigated the chemical reactivity of the metabolically activated forms of ArNH₂, that is, ArNHOH and ArNHOAc, toward 9-methylguanine by DFT calculations. The chemical reactivity of these forms is determined by the rate constants of two consecutive reactions leading to cationic guanine intermediates. The formation of ArNH⁺ accelerates with resonance stabilization of ArNH⁺, whereas the formed ArNH⁺ reacts with guanine derivatives with the constant diffusion-limited rate until the reaction slows down when ArNH⁺ is about 20 kcal/mol more stable than PhNH⁺. At this point, ArNHOH and ArNHOAc show maximum reactivity. The lowest activation energy of the reaction of ArNH⁺ with 9-methylguanine corresponds to the charge-transfer π-stacked transition state (π-TS) that leads to the direct formation of the C8 intermediate. The predicted activation barriers of this reaction match the observed absolute rate constants for a number of ArNH⁺. We demonstrate that the mutagenic potency of ArNH₂ correlates with the rate of formation and the chemical reactivity of the metabolically activated forms toward the C8 atom of dG. On the basis of geometric consideration of the π-TS complex made of genotoxic compounds with long aromatic systems, we propose that precovalent intercalation in DNA is not an essential step in the genotoxicity pathway of ArNH₂. The mechanism-based reasoning suggests rational design strategies to

  19. 78 FR 3900 - Generic Drug User Fee-Active Pharmaceutical Ingredient and Finished Dosage Form Facility Fee...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-17

    ... HUMAN SERVICES Food and Drug Administration Generic Drug User Fee--Active Pharmaceutical Ingredient and... drug active pharmaceutical ingredient (API) and finished dosage form (FDF) facilities user fees for... applications in the backlog as of October 1, 2012, on finished dosage form (FDF) and active...

  20. Parieto-frontal gamma band activity during the perceptual emergence of speech forms.

    PubMed

    Basirat, Anahita; Sato, Marc; Schwartz, Jean-Luc; Kahane, Philippe; Lachaux, Jean-Philippe

    2008-08-01

    The multistable perception of speech refers to the perceptual changes experienced while listening to a speech form cycled in rapid and continuous repetition, the so-called Verbal Transformation Effect. Because distinct interpretations of the same repeated stimulus alternate spontaneously, this effect provides an invaluable tool to examine how speech percepts are formed in the listener's mind. In order to track the temporal dynamics of brain activity specifically linked to perceptual changes, intracerebral EEG activity was recorded from two implanted epileptic patients while performing a verbal transformation task. To this aim, they were asked to carefully listen to a speech sequence played repeatedly and to press a button whenever they perceived a change in the repeated utterance. For both patients, 300-800 ms prior to the reported perceptual transitions, high frequency activity in the gamma band range (>40 Hz) was observed within the left inferior frontal and supramarginal gyri. An additional auditory decision task was used to rule out the possibility that the increased gamma band activity was due to the patients' motor responses. These results suggest that articulatory-based representations play a key part in the endogenously driven emergence of auditory speech percepts. The findings are interpreted in relation to theories assuming a link between perception and action in the human speech processing system.

  1. The Effects of Two Forms of Physical Activity on Eyeblink Classical Conditioning

    PubMed Central

    Green, John T.; Chess, Amy C.; Burns, Montana; Schachinger, Kira M.; Thanellou, Alexandra

    2011-01-01

    Voluntary exercise, in the form of free access to a running wheel in the home cage, has been shown to improve several forms of learning and memory. Acrobatic training, in the form of learning to traverse an elevated obstacle course, has been shown to induce markers of neural plasticity in the cerebellar cortex in rodents. In three experiments, we examined the effects of these two forms of physical activity on delay eyeblink conditioning in rats. In Experiment 1, exercising rats were given 17 days of free access to a running wheel in their home cage prior to 10 days of delay eyeblink conditioning. Rats that exercised conditioned significantly better and showed a larger reflexive eyeblink unconditioned response to the periocular stimulation unconditioned stimulus than rats that did not exercise. In Experiment 2, exercising rats were given 17 days of free access to a running wheel in their home cage prior to 10 days of explicitly unpaired stimulus presentations. Rats that exercised responded the same to tone, light, and periocular stimulation as rats that did not exercise. In Experiment 3, acrobatic training rats were given 15 days of daily training on an elevated obstacle course prior to 10 days of eyeblink conditioning. Activity control rats underwent 15 days of yoked daily running in an open field. Rats that underwent acrobatic training did not differ in eyeblink conditioning from activity control rats. The ability to measure the learned response precisely, and the well-mapped neural circuitry of eyeblink conditioning offer some advantages for the study of exercise effects on learning and memory. PMID:21238502

  2. Multifunctional porous titanium oxide coating with apatite forming ability and photocatalytic activity on a titanium substrate formed by plasma electrolytic oxidation.

    PubMed

    Akatsu, T; Yamada, Y; Hoshikawa, Y; Onoki, T; Shinoda, Y; Wakai, F

    2013-12-01

    Plasma electrolytic oxidation (PEO) was used to make a multifunctional porous titanium oxide (TiO2) coating on a titanium substrate. The key finding of this study is that a highly crystalline TiO2 coating can be made by performing the PEO in an ammonium acetate (CH3COONH4) solution; the PEO coating was formed by alternating between rapid heating by spark discharges and quenching in the solution. The high crystallinity of the TiO2 led to the surface having multiple functions, including apatite forming ability and photocatalytic activity. Hydroxyapatite formed on the PEO coating when it was soaked in simulated body fluid. The good apatite forming ability can be attributed to the high density of hydroxyl groups on the anatase and rutile phases in the coating. The degradation of methylene blue under ultraviolet radiation indicated that the coating had high photocatalytic activity.

  3. A comparison of the efficiency of G protein activation by ligand-free and light-activated forms of rhodopsin.

    PubMed Central

    Melia, T J; Cowan, C W; Angleson, J K; Wensel, T G

    1997-01-01

    Activation of the photoreceptor G protein transducin (Gt) by opsin, the ligand-free form of rhodopsin, was measured using rod outer segment membranes with densities of opsin and Gt similar to those found in rod cells. When GTPgammaS was used as the activating nucleotide, opsin catalyzed transducin activation with an exponential time course with a rate constant k(act) on the order of 2 x 10(-3)s(-1). Comparison under these conditions to activation by flash-generated metarhodopsin II (MII) revealed that opsin- and R*-catalyzed activation showed similar kinetics when MII was present at a surface density approximately 10(-6) lower than that of opsin. Thus, in contrast to some previous reports, we find that the catalytic potency of opsin is only approximately 10(-6) that of MII. In the presence of residual retinaldehyde-derived species present in membranes treated with hydroxylamine after bleaching, the apparent k(act) observed was much higher than that for opsin, suggesting a possible explanation for previous reports of more efficient activation by opsin. These results are important for considering the possible role of opsin in the diverse phenomena in which it has been suggested to play a key role, such as bleaching desensitization and retinal degeneration induced by continuous light or vitamin A deprivation. PMID:9414230

  4. Special Form Testing of Sealed Source Encapsulation for High-Alpha-Activity Actinide Materials

    SciTech Connect

    Martinez, Oscar A

    2016-01-01

    In the United States all transportation of radioactive material is regulated by the U.S. Department of Transportation (DOT). Beginning in 2008 a new type of sealed-source encapsulation package was developed and tested by Oak Ridge National Laboratory (ORNL). These packages contain high-alpha-activity actinides and are regulated and transported in accordance with the requirements for DOT Class 7 hazardous material. The DOT provides specific regulations pertaining to special form encapsulation designs. The special form designation indicates that the encapsulated radioactive contents have a very low probability of dispersion even when subjected to significant structural events. The special form designs have been shown to simplify the delivery, transport, acceptance, and receipt processes. It is intended for these sealed-source encapsulations to be shipped to various facilities making it very advantageous for them to be certified as special form. To this end, DOT Certificates of Competent Authority (CoCAs) have been sought for the design suitable for containing high-alpha-activity actinide materials. This design consists of the high-alpha-activity material encapsulated within a triangular zirconia canister, referred to as a ZipCan, tile that is then enclosed by a spherical shell. The spherical shell design, with ZipCan tile inside, was tested for compliance with the special form regulations found in 49 CFR 173.469. The spherical enclosure was subjected to 9-m impact, 1 m percussion, and 10-minute thermal tests at the Packaging Evaluation Facility located at the National Transportation Research Center in Knoxville, TN USA and operated by ORNL. Before and after each test, the test units were subjected to a helium leak check and a bubble test. The ZipCan tiles and core were also subjected to the tests required for ISO 2919:2012(E), including a Class IV impact test and heat test and subsequently subjected to helium leakage rate tests [49 CFR 173.469(a)(4)(i)]. The impact

  5. Staphylococcus aureus forms spreading dendrites that have characteristics of active motility

    PubMed Central

    Pollitt, Eric J. G.; Crusz, Shanika A.; Diggle, Stephen P.

    2015-01-01

    Staphylococcus aureus is historically regarded as a non-motile organism. More recently it has been shown that S. aureus can passively move across agar surfaces in a process called spreading. We re-analysed spreading motility using a modified assay and focused on observing the formation of dendrites: branching structures that emerge from the central colony. We discovered that S. aureus can spread across the surface of media in structures that we term ‘comets’, which advance outwards and precede the formation of dendrites. We observed comets in a diverse selection of S. aureus isolates and they exhibit the following behaviours: (1) They consist of phenotypically distinct cores of cells that move forward and seed other S. aureus cells behind them forming a comet ‘tail’; (2) they move when other cells in the comet tail have stopped moving; (3) the comet core is held together by a matrix of slime; and (4) the comets etch trails in the agar as they move forwards. Comets are not consistent with spreading motility or other forms of passive motility. Comet behaviour does share many similarities with a form of active motility known as gliding. Our observations therefore suggest that S. aureus is actively motile under certain conditions. PMID:26680153

  6. An Aqueous Extract of Marine Microalgae Exhibits Antimetastatic Activity through Preferential Killing of Suspended Cancer Cells and Anticolony Forming Activity

    PubMed Central

    Somasekharan, Syam Prakash; El-Naggar, Amal; Sorensen, Poul H.

    2016-01-01

    Research on marine natural products as potential anticancer agents is still limited. In the present study, an aqueous extract of a Canadian marine microalgal preparation was assessed for anticancer activities using various assays and cell lines of human cancers, including lung, prostate, stomach, breast, and pancreatic cancers, as well as an osteosarcoma. In vitro, the microalgal extract exhibited marked anticolony forming activity. In addition, it was more toxic, as indicated by increased apoptosis, to nonadherent cells (grown in suspension) than to adherent cells. In vivo, an antimetastatic effect of the extract was observed in NOD-SCID mice carrying subrenal capsule xenografts of PC3 prostate cancer cells. The results of the present study suggest that the antimetastatic effect of the aqueous microalgal extract is based on inhibition of colony forming ability of cancer cells and the preferential killing of suspended cancer cells. Further research aimed at identification of the molecular basis of the anticancer activities of the microalgal extract appears to be warranted. PMID:27656243

  7. Redox-sensitive extracellular gates formed by auxiliary beta subunits of calcium-activated potassium channels.

    PubMed

    Zeng, Xu-Hui; Xia, Xiao-Ming; Lingle, Christopher J

    2003-06-01

    An important step to understanding ion channels is identifying the structural components that act as the gates to ion movement. Here we describe a new channel gating mechanism, produced by the beta3 auxiliary subunits of Ca2+-activated, large-conductance BK-type K+ channels when expressed with their pore-forming alpha subunits. BK beta subunits have a cysteine-rich extracellular segment connecting two transmembrane segments, with small cytosolic N and C termini. The extracellular segments of the beta3 subunits form gates to block ion permeation, providing a mechanism by which current can be rapidly diminished upon cellular repolarization. Furthermore, this gating mechanism is abolished by reduction of extracellular disulfide linkages, suggesting that endogenous mechanisms may regulate this gating behavior. The results indicate that auxiliary beta subunits of BK channels reside sufficiently close to the ion permeation pathway defined by the alpha subunits to influence or block access of small molecules to the permeation pathway.

  8. Antifungal activity and pore-forming mechanism of astacidin 1 against Candida albicans.

    PubMed

    Choi, Hyemin; Lee, Dong Gun

    2014-10-01

    In a previous report, a novel antibacterial peptide astacidin 1 (FKVQNQHGQVVKIFHH) was isolated from hemocyanin of the freshwater crayfish Pacifastacus leniusculus. In this study, the antifungal activity and mechanism of astacidin 1 were evaluated. Astacidin 1 exhibited antifungal activity against Candida albicans, Trichosporon beigelii, Malassezia furfur, and Trichophyton rubrum. Also, astacidin 1 had fungal cell selectivity in human erythrocytes without causing hemolysis. To understand the antifungal mechanism, membrane studies were done against C. albicans and T. beigelii. Flow cytometric analysis and K(+) measurement showed membrane damage, resulting in membrane permeabilization and K(+) release-induced membrane depolarization. Furthermore, the calcein leakage from liposomes mimicking C. albicans membrane demonstrated that the membrane-active action was driven by pore-forming mechanism. Live cell imaging using fluorescein isothiocyanate-labeled dextrans of various sizes suggested that the radii of pores formed in the C. albicans membrane were 1.4-2.3 nm. Therefore, the present study suggests that astacidin 1 exerts its antifungal effect by damaging the fungal membrane via pore formation.

  9. Activated platelets form protected zones of adhesion on fibrinogen and fibronectin-coated surfaces

    PubMed Central

    1993-01-01

    Leukocytes form zones of close apposition when they adhere to ligand- coated surfaces. Because plasma proteins are excluded from these contact zones, we have termed them protected zones of adhesion. To determine whether platelets form similar protected zones of adhesion, gel-filtered platelets stimulated with thrombin or ADP were allowed to adhere to fibrinogen- or fibronectin-coated surfaces. The protein- coated surfaces with platelets attached were stained with either fluorochrome-conjugated goat anti-human fibrinogen or anti-human fibronectin antibodies, or with rhodamine-conjugated polyethylene glycol polymers. Fluorescence microscopy revealed that F(ab')2 anti- fibrinogen (100 kD) did not penetrate into the contact zones between stimulated platelets and the underlying fibrinogen-coated surface, while Fab antifibrinogen (50 kD) and 10 kD polyethylene glycol readily penetrated and stained the substrate beneath the platelets. Thrombin- or ADP-stimulated platelets also formed protected zones of adhesion on fibronectin-coated surfaces. F(ab')2 anti-fibronectin and 10 kD polyethylene glycol were excluded from these adhesion zones, indicating that they are much less permeable than those formed by platelets on fibrinogen-coated surfaces. The permeability properties of protected zones of adhesion formed by stimulated platelets on surfaces coated with both fibrinogen and fibronectin were similar to the zones of adhesion formed on fibronectin alone. mAb 7E3, directed against the alpha IIb beta 3 integrin blocked the formation of protected adhesion zones between thrombin-stimulated platelets and fibrinogen or fibronectin coated surfaces. mAb C13 is directed against the alpha 5 beta 1 integrin on platelets. Stimulated platelets treated with this mAb formed protected zones of adhesion on surfaces coated with fibronectin. These protected zones were impermeable to F(ab')2 antifibronectin but were permeable to 10 kD polyethylene glycol. These results show that activated

  10. [Suppression of telomerase activity leukemic cells by mutant forms of Rhodospirillum rubrum L-asparaginase].

    PubMed

    Pokrovskaya, M V; Zhdanov, D D; Eldarov, M A; Aleksandrova, S S; Veselovskiy, A V; Pokrovskiy, V S; Grishin, D V; Gladilina, Ju A; Sokolov, N N

    2017-01-01

    The active and stable mutant forms of short chain cytoplasmic L-asparaginase type I of Rhodospirillum rubrum (RrA): RrA+N17, D60K, F61L, RrA+N17, A64V, E67K, RrA+N17, E149R, V150P, RrAE149R, V150P and RrAE149R, V150P, F151T were obtained by the method of site-directed mutagenesis. It is established that variants RrA-N17, E149R, V150P, F151T and RrАE149R, V150P are capable to reduce an expression hTERT subunit of telomerase and, hence, activity of telomeres in Jurkat cells, but not in cellular lysates. During too time, L-asparaginases of Escherichia coli, Erwinia carotovora and Wolinella succinogenes, mutant forms RrА+N17, D60K, F61L and RrА+N17, A64V, E67K do not suppress of telomerase activity. The assumption of existence in structure RrA of areas (amino acids residues in the position 146-164, 1-17, 60-67) which are responsible for suppression of telomerase activity is made. The received results show that antineoplastic activity of some variants RrA is connected both with reduction of concentration of free L-asparagine, and with expression suppression of hTERT telomerase subunit, that opens new prospects for antineoplastic therapy.

  11. A Natural Chimeric Pseudomonas Bacteriocin with Novel Pore-Forming Activity Parasitizes the Ferrichrome Transporter

    PubMed Central

    Kemland, Lieselore; Anoz-Carbonell, Ernesto; Buchanan, Susan K.; De Mot, René

    2017-01-01

    ABSTRACT Modular bacteriocins represent a major group of secreted protein toxins with a narrow spectrum of activity, involved in interference competition between Gram-negative bacteria. These antibacterial proteins include a domain for binding to the target cell and a toxin module at the carboxy terminus. Self-inhibition of producers is provided by coexpression of linked immunity genes that transiently inhibit the toxin’s activity through formation of bacteriocin-immunity complexes or by insertion in the inner membrane, depending on the type of toxin module. We demonstrate strain-specific inhibitory activity for PmnH, a Pseudomonas bacteriocin with an unprecedented dual-toxin architecture, hosting both a colicin M domain, potentially interfering with peptidoglycan synthesis, and a novel colicin N-type domain, a pore-forming module distinct from the colicin Ia-type domain in Pseudomonas aeruginosa pyocin S5. A downstream-linked gene product confers PmnH immunity upon susceptible strains. This protein, ImnH, has a transmembrane topology similar to that of Pseudomonas colicin M-like and pore-forming immunity proteins, although homology with either of these is essentially absent. The enhanced killing activity of PmnH under iron-limited growth conditions reflects parasitism of the ferrichrome-type transporter for entry into target cells, a strategy shown here to be used as well by monodomain colicin M-like bacteriocins from pseudomonads. The integration of a second type of toxin module in a bacteriocin gene could offer a competitive advantage against bacteria displaying immunity against only one of both toxic activities. PMID:28223456

  12. Activation of large form galanin-LI by extracellular processing in small cell lung carcinoma tissue.

    PubMed

    Yamamoto, Hiroyuki; Iguchi, Kazuaki; Ohno, Satoshi; Yokogawa, Takashi; Nishikawa, Kazuya; Hoshino, Minoru

    2011-10-01

    Galanin is a neuropeptide that is widely distributed in the central and peripheral nervous systems. Some small cell lung carcinoma (SCLC) cell lines such as SBC-3A release only the high-molecular-mass form, with lower molecular mass forms being undetectable. To investigate the mechanism of processing of progalanin to active peptide, we studied galanin-LI in both the culture media of SBC-3A cells and in extracts from in vivo mouse SBC-3A tumors. SBC-3A cells were found to release high molecular mass galanin, but did not release active peptides. In contrast, tumor extract contained both high-molecular-mass galanin, and a cleaved lower-molecular-mass form of the peptide (8, 5 and 2 kDa). The lower-molecular-mass peptide was identified as galanin(1-20) by MALDI-TOF mass spectrometry. We then looked at MMP-2 and MMP-9 release from SBC-3A cells and tumor tissue treated with galanin and progalanin, as revealed by gelatin zymography. Galanin elicited pro-MMP-2 and pro-MMP-9 release from SBC-3A cells and tumor tissue; however, recombinant progalanin induced pro-MMP-2 and pro-MMP-9 release from tumor tissue only. This study has shown that the galanin-LI released from SCLC SBC-3A cells consisted of the high-molecular-mass peptide form, and was processed extracellularly to galanin(1-20). Furthermore, galanin was seen to induce pro-MMP-2 and pro-MMP-9 release from SBC-3A cells.

  13. Channel-forming activity in the venom of the cockroach-hunting wasp, Ampulex compressa.

    PubMed

    Gincel, Dan; Haspel, Gal; Libersat, Frederic

    2004-05-01

    The parasitoid solitary wasp Ampulex compressa uses the cockroach Periplaneta americana as a food supply for its larvae. To subdue its prey, the wasp injects a venom cocktail into the brain of the cockroach. We investigated channel activity of A. compressa venom by collecting venom and incorporating it into a planar lipid bilayer. The venom, reconstituted into the bilayer, showed ion channel activity, forming a fast-fluctuating channel with a small conductance of 20+/-0.1pS, with no voltage sensitivity. These channels were not observed when the venom was digested with proteases before application to the bilayer, but were not affected by exposure to protease after their incorporation into the bilayer, indicating that the active venom component is a peptide. The channels were found to be cation selective with similar selectivity for the monovalent cations K(+), Li(+) and Na(+), but showed high selectivity against anions (Cl(-)) and divalent cations (Ca(2+) and Mg(2+)). This study is the first demonstration and biophysical characterization of channel activity in the venom of A. compressa. The possible functional significance of this channel activity is discussed in light of the unusual nature of the effects of this wasp venom on the behavior of its prey.

  14. Stress and host immunity amplify Mycobacterium tuberculosis phenotypic heterogeneity and induce nongrowing metabolically active forms.

    PubMed

    Manina, Giulia; Dhar, Neeraj; McKinney, John D

    2015-01-14

    Nonreplicating and metabolically quiescent bacteria are implicated in latent tuberculosis infections and relapses following "sterilizing" chemotherapy. However, evidence linking bacterial dormancy and persistence in vivo is largely inconclusive. Here we measure the single-cell dynamics of Mycobacterium tuberculosis replication and ribosomal activity using quantitative time-lapse microscopy and a reporter of ribosomal RNA gene expression. Single-cell dynamics exhibit heterogeneity under standard growth conditions, which is amplified by stressful conditions such as nutrient limitation, stationary phase, intracellular replication, and growth in mouse lungs. Additionally, the lungs of chronically infected mice harbor a subpopulation of nongrowing but metabolically active bacteria, which are absent in mice lacking interferon-γ, a cytokine essential for antituberculosis immunity. These cryptic bacterial forms are prominent in mice treated with the antituberculosis drug isoniazid, suggesting a role in postchemotherapeutic relapses. Thus, amplification of bacterial phenotypic heterogeneity in response to host immunity and drug pressure may contribute to tuberculosis persistence.

  15. A Disintegrin and Metalloproteinase with Thrombospondin Motifs-5 (ADAMTS-5) Forms Catalytically Active Oligomers*

    PubMed Central

    Kosasih, Hansen J.; Last, Karena; Rogerson, Fraser M.; Golub, Suzanne B.; Gauci, Stephanie J.; Russo, Vincenzo C.; Stanton, Heather; Wilson, Richard; Lamande, Shireen R.; Holden, Paul; Fosang, Amanda J.

    2016-01-01

    The metalloproteinase ADAMTS-5 (A disintegrin and metalloproteinase with thrombospondin motifs) degrades aggrecan, a proteoglycan essential for cartilage structure and function. ADAMTS-5 is the major aggrecanase in mouse cartilage, and is also likely to be the major aggrecanase in humans. ADAMTS-5 is a multidomain enzyme, but the function of the C-terminal ancillary domains is poorly understood. We show that mutant ADAMTS-5 lacking the catalytic domain, but with a full suite of ancillary domains inhibits wild type ADAMTS activity, in vitro and in vivo, in a dominant-negative manner. The data suggest that mutant ADAMTS-5 binds to wild type ADAMTS-5; thus we tested the hypothesis that ADAMTS-5 associates to form oligomers. Co-elution, competition, and in situ PLA experiments using full-length and truncated recombinant ADAMTS-5 confirmed that ADAMTS-5 molecules interact, and showed that the catalytic and disintegrin-like domains support these intermolecular interactions. Cross-linking experiments revealed that recombinant ADAMTS-5 formed large, reduction-sensitive oligomers with a nominal molecular mass of ∼400 kDa. The oligomers were unimolecular and proteolytically active. ADAMTS-5 truncates comprising the disintegrin and/or catalytic domains were able to competitively block full-length ADAMTS-5-mediated aggrecan cleavage, measured by production of the G1-EGE373 neoepitope. These results show that ADAMTS-5 oligomerization is required for full aggrecanase activity, and they provide evidence that blocking oligomerization inhibits ADAMTS-5 activity. The data identify the surface provided by the catalytic and disintegrin-like domains of ADAMTS-5 as a legitimate target for the design of aggrecanase inhibitors. PMID:26668318

  16. The membrane protein Pannexin1 forms two open channel conformations depending on the mode of activation

    PubMed Central

    Wang, Junjie; Ambrosi, Cinzia; Qiu, Feng; Jackson, David G.; Sosinsky, Gina; Dahl, Gerhard

    2014-01-01

    Pannexin1 (Panx1) participates in several signaling events that involve ATP release, including the innate immune response, ciliary beat in airway epithelia and oxygen supply in the vasculature. The view that Panx1 forms a large ATP-release channel has been challenged by the association of a low conductance, small anion-selective channel with the presence of Panx1. We showed that Panx1 membrane channels can function in two distinct modes with different conductances and permeabilities when heterologously expressed in Xenopus oocytes. When stimulated by potassium ions (K+), Panx1 formed a high conductance channel of ~500 pS that was permeable to ATP. Various physiological stimuli can induce this ATP-permeable conformation of the channel in several cell types. In contrast, the channel had a low conductance (~50 pS) with no detectable ATP permeability when activated by voltage in the absence of K+. The two channel states were associated with different reactivities of the terminal cysteine of Panx1 to thiol reagents, suggesting different conformations. Single particle electron microscopic analysis revealed that K+ stimulated the formation of channels with a larger pore diameter than those formed in the absence of K+. These data suggest that different stimuli lead to distinct channel structures with distinct biophysical properties. PMID:25056878

  17. Trypanocidal activity of the essential oils in their conventional and nanoemulsion forms: in vitro tests.

    PubMed

    Baldissera, Matheus D; Da Silva, Aleksandro S; Oliveira, Camila B; Zimmermann, Carine E P; Vaucher, Rodrigo A; Santos, Roberto C V; Rech, Virginia C; Tonin, Alexandre A; Giongo, Janice L; Mattos, Cristiane B; Koester, Letícia; Santurio, Janio M; Monteiro, Silvia G

    2013-07-01

    The aim of this study was to investigate the susceptibility in vitro of Trypanosoma evansi to the essential oils of andiroba (Carapa guaianensis) and aroeira (Schinus molle), in their conventional and nanostructured forms. For that, pure oils at concentrations of 0.5%, 1.0% and 2.0% were used. A negative control (untreated) and a positive control (diminazene aceturate 0.5%) were used as comparative parameters. Later, the same tests were performed, using nanoemulsions oils at concentrations of 0.5% and 1.0%. The tests were carried out in triplicates and the numbers of parasites were quantified on 1, 3 and 6 h from onset of the study. A dose-dependent reduction in the number of parasites to the forms of two oils tested was observed after 1 h. The concentration of parasites was significantly reduced at low concentrations after 3 h, as well as at 6 h no alive parasites were observed for the essential oils tested. Ours findings indicate, for the first time, that oils of andiroba and aroeira (in their conventional and nanoemulsion forms) have high activity against T. evansi in vitro, leading to the suggestion that these oils may be applied as an alternative treatment for this disease.

  18. Hydroxyapatite formed on/in agarose gel induces activation of blood coagulation and platelets aggregation.

    PubMed

    Arimura, Shin-ichiro; Kawahara, Ko-ichi; Biswas, Kamal Krishna; Abeyama, Kazuhiro; Tabata, Masashi; Shimoda, Toru; Ogomi, Daisuke; Matsusaki, Michiya; Kato, Shinya; Ito, Takashi; Sugihara, Kazumasa; Akashi, Mitsuru; Hashiguchi, Teruto; Maruyama, Ikuro

    2007-05-01

    We reported earlier that hydroxyapatite (HA) formed on/in agarose gels (HA/agarose) produced by alternate soaking process is a bone-filling material possessing osteoconductive and hemostatic effects. This process could allow us to make bone-like apatite that was formed on/in organic polymer hydrogel matrices. Here, we investigated the mechanism of hemostasis induced by HA/agarose and found that HA/agarose, but not agarose or HA powder, significantly shortened activated partial thromboplastin time (APTT). While HA/agarose did not show significant platelet aggregation, it markedly enhanced adenosine diphosphate (ADP)-induced platelet aggregation. Moreover, Western blot analysis revealed selective adsorption of vitronectin onto HA/agarose. We also observed marked differences between HA powder and HA/agarose in their XRD patterns. The crystallinity of HA powder was much higher compared to that of HA/agarose. Furthermore, 50-100 nm of tube-form aggregations was observed in HA powder on the other hand 100-200 nm of particles was observed in HA/agarose by SEM observation. Thus 100-200 nm of low crystallized particles on the surface structure of HA/agarose may play an important role in hemostasis. Our results demonstrated a crucial role of HA/agarose in the mechanism of hemostasis and suggested a potential role for HA/agarose as a bone-grafting material.

  19. The TCP1γ subunit of Leishmania donovani forms a biologically active homo-oligomeric complex.

    PubMed

    Bhaskar; Mitra, Kalyan; Kuldeep, Jitendra; Siddiqi, Mohammad Imran; Goyal, Neena

    2015-12-01

    Chaperonins are a class of molecular chaperons that encapsulate nascent or stress-denatured proteins and assist their intracellular assembly and folding in an ATP-dependent manner. The ubiquitous eukaryotic chaperonin, TCP1 ring complex is a hetero-oligomeric complex comprising two rings, each formed of eight subunits that may have distinct substrate recognition and ATP hydrolysis properties. In Leishmania, only the TCP1γ subunit has been cloned and characterized. It exhibited differential expression at various growth stages of promastigotes. In the present study, we expressed the TCP1γ subunit in Escherichia coli to investigate whether it forms chaperonin-like complexes and plays a role in protein folding. LdTCP1γ formed high-molecular-weight complexes within E. coli cells as well as in Leishmania cell lysates. The recombinant protein is arranged into two back-to-back rings of seven subunits each, as predicted by homology modelling and observed by negative staining electron microscopy. This morphology is consistent with that of the oligomeric double-ring group I chaperonins found in mitochondria. The LdTCP1γ homo-oligomeric complex hydrolysed ATP, and was active as assayed by luciferase refolding. Thus, the homo-oligomer performs chaperonin reactions without partner subunit(s). Further, co-immunoprecipitation studies revealed that LdTCP1γ interacts with actin and tubulin proteins, suggesting that the complex may have a role in maintaining the structural dynamics of the cytoskeleton of parasites.

  20. Theoretical study of the phototoxicity of naproxen and the active form of nabumetone.

    PubMed

    Musa, Klefah A K; Eriksson, Leif A

    2008-10-30

    Density functional theory using the hybrid functional B3LYP has been employed in order to study the mechanisms of photoinduced decomposition of the closely related nonsteroidal anti-inflammatory drugs naproxen (NP) and 6-methoxy-2-naphthylacetic acid (MNAA; the active form of nabumetone). The photochemical properties and computed energies of various species obtained in this study show that both drugs dominate in their deprotonated forms at physiological pH. The deprotonated acids are unable to decarboxylate from their excited singlets; instead, they decarboxylate from their first excited triplet states with high efficiency, overcoming energy barriers less than 3 and 1 kcal/mol for MNAA and NP, respectively. The ultraviolet and visible spectra of the neutral, deprotonated, and decarboxylated moieties of MNAA and NP are more-or-less similar but with higher probabilites (oscillator strength) for the latter. This fact, as well as the higher reactivity of NP, is explained in terms of the electron-donating effect of the additional methyl group present in NP. Singlet oxygen, superoxide radical anion, and corresponding peroxyl radical species are expected to be formed in different steps throughout the proposed photodegradation pathways of both drugs, which give rise to their effects on biomolecules, for example, lipid peroxidation.

  1. Inter-molecular crosslinking activity is engendered by the dimeric form of transglutaminase 2.

    PubMed

    Kim, Nayeon; Lee, Won-Kyu; Lee, Seon-Hyeong; Jin, Kyeong Sik; Kim, Kyung-Hee; Lee, Younho; Song, Minsoo; Kim, Soo-Youl

    2017-03-01

    Transglutaminase 2 (TGase 2) catalyzes a crosslink between protein bound-glutamine and -lysine. We proposed the mechanism of TGase 2 activation depends on conformation change from unfolded monomer to unfolded dimer. We found that TGase 2 has temperature-sensitive conformation change system at 30 °C. Small-angle X-ray scattering analysis showed that the enzyme was maintained as an unfolded monomer at temperatures below 30 °C, but changed to an unfolded dimer at over 30 °C. Mass analysis revealed that the C-terminus of TGase 2 was the critical region for dimerization. Furthermore, this conformational switch creates new biochemical reactivity that catalyzed inter-molecular crosslink at above 30 °C as an unfolded dimer of TGase 2 while catalyzed intra-molecular crosslink at below 30 °C as an unfolded monomer of TGase 2. The mechanism of TGase 2 activation depends on temperature-sensitive conformation change from unfolded monomer to unfolded dimer at over 30 °C. Furthermore, inter-molecular crosslinking activity is generated by the dimeric form of TGase 2. TGase 2 switches its conformation from a monomer to a dimer following a change in temperature, which engendered unique catalytic function of enzyme as inter-molecular crosslinking activity with calcium.

  2. Prospective study of urban form and physical activity in the Black Women's Health Study.

    PubMed

    Coogan, Patricia F; White, Laura F; Adler, Thomas J; Hathaway, Kevin M; Palmer, Julie R; Rosenberg, Lynn

    2009-11-01

    The authors used data from the Black Women's Health Study to assess the association between neighborhood urban form and physical activity. Women reported hours/week of utilitarian and exercise walking and of vigorous activity in 1995 and on biennial follow-up questionnaires through 2001. Housing density, road networks, availability of public transit, sidewalks, and parks were characterized for the residential neighborhoods of 20,354 Black Women's Health Study participants living in New York, New York; Chicago, Illinois; and Los Angeles, California. The authors quantified the associations between features of the environment and physical activity using odds ratios for >or=5 relative to <5 hours/week of physical activity. For all women, housing density had the strongest association with utilitarian walking (odds ratio for the most- compared with the least-dense quintile = 2.72, 95% confidence interval: 2.22, 3.31), followed by availability of public transit. Women who moved during follow-up to neighborhoods of lower density were 36% more likely to decrease their levels of utilitarian walking, and those who moved to neighborhoods of higher density were 23% more likely to increase their levels of utilitarian walking, relative to women who moved to neighborhoods of similar density. These data suggest that increases in housing density may lead to increases in utilitarian walking among African-American women.

  3. 77 FR 23735 - Agency Information Collection Activities: Forms G-1041 and G-1041A, Extension of a Currently...

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  4. 77 FR 64388 - Agency Information Collection (Former POW Medical History), VA Form 10-0048 Activities Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-19

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  5. 75 FR 65499 - Agency Information Collection Activities: Form I-600/I-600A, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

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  6. 75 FR 35824 - Agency Information Collection Activities: Forms I-600/I-600A, Extension of a Currently Approved...

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    2010-06-23

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  7. Active subtilisin-like protease from a hyperthermophilic archaeon in a form with a putative prosequence.

    PubMed

    Kannan, Y; Koga, Y; Inoue, Y; Haruki, M; Takagi, M; Imanaka, T; Morikawa, M; Kanaya, S

    2001-06-01

    The gene encoding subtilisin-like protease T. kodakaraensis subtilisin was cloned from a hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. T. kodakaraensis subtilisin is a member of the subtilisin family and composed of 422 amino acid residues with a molecular weight of 43,783. It consists of a putative presequence, prosequence, and catalytic domain. Like bacterial subtilisins, T. kodakaraensis subtilisin was overproduced in Escherichia coli in a form with a putative prosequence in inclusion bodies, solubilized in the presence of 8 M urea, and refolded and converted to an active molecule. However, unlike bacterial subtilisins, in which the prosequence was removed from the catalytic domain by autoprocessing upon refolding, T. kodakaraensis subtilisin was refolded in a form with a putative prosequence. This refolded protein of recombinant T. kodakaraensis subtilisin which is composed of 398 amino acid residues (Gly(-82) to Gly(316)), was purified to give a single band on a sodium dodecyl sulfate (SDS)-polyacrylamide gel and characterized for biochemical and enzymatic properties. The good agreement of the molecular weights estimated by SDS-polyacrylamide gel electrophoresis (44,000) and gel filtration (40,000) suggests that T. kodakaraensis subtilisin exists in a monomeric form. T. kodakaraensis subtilisin hydrolyzed the synthetic substrate N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide only in the presence of the Ca(2+) ion with an optimal pH and temperature of pH 9.5 and 80 degrees C. Like bacterial subtilisins, it showed a broad substrate specificity, with a preference for aromatic or large nonpolar P1 substrate residues. However, it was much more stable than bacterial subtilisins against heat inactivation and lost activity with half-lives of >60 min at 80 degrees C, 20 min at 90 degrees C, and 7 min at 100 degrees C.

  8. 12 CFR Appendix to Part 1253 - Prior Approval for Enterprise Products-Instructions and Notice of New Activity Form

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Approval for Enterprise Products—Instructions and Notice of New Activity Form ER02JY09.000 ER02JY09.001... 12 Banks and Banking 7 2010-01-01 2010-01-01 false Prior Approval for Enterprise Products-Instructions and Notice of New Activity Form Appendix to Part 1253 Banks and Banking FEDERAL HOUSING...

  9. Biofilm-forming activity of bacteria isolated from toilet bowl biofilms and the bactericidal activity of disinfectants against the isolates.

    PubMed

    Mori, Miho; Gomi, Mitsuhiro; Matsumune, Norihiko; Niizeki, Kazuma; Sakagami, Yoshikazu

    2013-01-01

    To evaluate the sanitary conditions of toilets, the bacterial counts of the toilet bowl biofilms in 5 Kansai area and 11 Kansai and Kanto area homes in Japan were measured in winter and summer seasons, respectively. Isolates (128 strains) were identified by analyzing 16S ribosomal RNA sequences. The number of colonies and bacterial species from biofilms sampled in winter tended to be higher and lower, respectively, than those in summer. Moreover, the composition of bacterial communities in summer and winter samples differed considerably. In summer samples, biofilms in Kansai and Kanto areas were dominated by Blastomonas sp. and Mycobacterium sp., respectively. Methylobacterium sp. was detected in all toilet bowl biofilms except for one sample. Methylobacterium sp. constituted the major presence in biofilms along with Brevundimonas sp., Sphingomonas sp., and/or Pseudomonas sp. The composition ratio of the sum of their genera was 88.0 from 42.9% of the total bacterial flora. The biofilm formation abilities of 128 isolates were investigated, and results suggested that Methylobacterium sp. and Sphingomonas sp. were involved in biofilm formation in toilet bowls. The biofilm formation of a mixed bacteria system that included bacteria with the highest biofilm-forming ability in a winter sample was greater than mixture without such bacteria. This result suggests that isolates possessing a high biofilm-forming activity are involved in the biofilm formation in the actual toilet bowl. A bactericidal test against 25 strains indicated that the bactericidal activities of didecyldimethylammonium chloride (DDAC) tended to be higher than those of polyhexamethylene biguanide (PHMB) and N-benzyl-N,N-dimethyldodecylammonium chloride (ADBAC). In particular, DDAC showed high bactericidal activity against approximately 90% of tested strains under the 5 h treatment.

  10. Extensive Post-translational Modification of Active and Inactivated Forms of Endogenous p53*

    PubMed Central

    DeHart, Caroline J.; Chahal, Jasdave S.; Flint, S. J.; Perlman, David H.

    2014-01-01

    The p53 tumor suppressor protein accumulates to very high concentrations in normal human fibroblasts infected by adenovirus type 5 mutants that cannot direct assembly of the viral E1B 55-kDa protein-containing E3 ubiquitin ligase that targets p53 for degradation. Despite high concentrations of nuclear p53, the p53 transcriptional program is not induced in these infected cells. We exploited this system to examine select post-translational modifications (PTMs) present on a transcriptionally inert population of endogenous human p53, as well as on p53 activated in response to etoposide treatment of normal human fibroblasts. These forms of p53 were purified from whole cell lysates by means of immunoaffinity chromatography and SDS-PAGE, and peptides derived from them were subjected to nano-ultra-high-performance LC-MS and MS/MS analyses on a high-resolution accurate-mass MS platform (data available via ProteomeXchange, PXD000464). We identified an unexpectedly large number of PTMs, comprising phosphorylation of Ser and Thr residues, methylation of Arg residues, and acetylation, ubiquitinylation, and methylation of Lys residues—for example, some 150 previously undescribed modifications of p53 isolated from infected cells. These modifications were distributed across all functional domains of both forms of the endogenous human p53 protein, as well as those of an orthologous population of p53 isolated from COS-1 cells. Despite the differences in activity, including greater in vitro sequence-specific DNA binding activity exhibited by p53 isolated from etoposide-treated cells, few differences were observed in the location, nature, or relative frequencies of PTMs on the two populations of human p53. Indeed, the wealth of PTMs that we have identified is consistent with a far greater degree of complex, combinatorial regulation of p53 by PTM than previously anticipated. PMID:24056736

  11. FUNCTION FOLLOWS FORM: ACTIVATION OF SHAPE & FUNCTION FEATURES DURING OBJECT IDENTIFICATION

    PubMed Central

    Yee, Eiling; Huffstetler, Stacy; Thompson-Schill, Sharon L.

    2011-01-01

    Most theories of semantic memory characterize knowledge of a given object as comprising a set of semantic features. But how does conceptual activation of these features proceed during object identification? We present the results of a pair of experiments that demonstrate that object recognition is a dynamically unfolding process in which function follows form. We used eye movements to explore whether activating one object’s concept leads to the activation of others that share perceptual (shape) or abstract (function) features. Participants viewed four-picture displays and clicked on the picture corresponding to a heard word. In critical trials, the conceptual representation of one of the objects in the display was similar in shape or function (i.e., its purpose) to the heard word. Importantly, this similarity was not apparent in the visual depictions (e.g., for the target “frisbee,” the shape-related object was a triangular slice of pizza – a shape that a frisbee cannot take); preferential fixations on the related object were therefore attributable to overlap of the conceptual representations on the relevant features. We observed relatedness effects for both shape and function, but shape effects occurred earlier than function effects. We discuss the implications of these findings for current accounts of the representation of semantic memory. PMID:21417543

  12. A Carboxyl Ester Lipase (CEL) Mutant Causes Chronic Pancreatitis by Forming Intracellular Aggregates That Activate Apoptosis.

    PubMed

    Xiao, Xunjun; Jones, Gabrielle; Sevilla, Wednesday A; Stolz, Donna B; Magee, Kelsey E; Haughney, Margaret; Mukherjee, Amitava; Wang, Yan; Lowe, Mark E

    2016-10-28

    Patients with chronic pancreatitis (CP) frequently have genetic risk factors for disease. Many of the identified genes have been connected to trypsinogen activation or trypsin inactivation. The description of CP in patients with mutations in the variable number of tandem repeat (VNTR) domain of carboxyl ester lipase (CEL) presents an opportunity to study the pathogenesis of CP independently of trypsin pathways. We tested the hypothesis that a deletion and frameshift mutation (C563fsX673) in the CEL VNTR causes CP through proteotoxic gain-of-function activation of maladaptive cell signaling pathways including cell death pathways. HEK293 or AR42J cells were transfected with constructs expressing CEL with 14 repeats in the VNTR (CEL14R) or C563fsX673 CEL (CEL maturity onset diabetes of youth with a deletion mutation in the VNTR (MODY)). In both cell types, CEL MODY formed intracellular aggregates. Secretion of CEL MODY was decreased compared with that of CEL14R. Expression of CEL MODY increased endoplasmic reticulum stress, activated the unfolded protein response, and caused cell death by apoptosis. Our results demonstrate that disorders of protein homeostasis can lead to CP and suggest that novel therapies to decrease the intracellular accumulation of misfolded protein may be successful in some patients with CP.

  13. Auditory selective attention to speech modulates activity in the visual word form area.

    PubMed

    Yoncheva, Yuliya N; Zevin, Jason D; Maurer, Urs; McCandliss, Bruce D

    2010-03-01

    Selective attention to speech versus nonspeech signals in complex auditory input could produce top-down modulation of cortical regions previously linked to perception of spoken, and even visual, words. To isolate such top-down attentional effects, we contrasted 2 equally challenging active listening tasks, performed on the same complex auditory stimuli (words overlaid with a series of 3 tones). Instructions required selectively attending to either the speech signals (in service of rhyme judgment) or the melodic signals (tone-triplet matching). Selective attention to speech, relative to attention to melody, was associated with blood oxygenation level-dependent (BOLD) increases during functional magnetic resonance imaging (fMRI) in left inferior frontal gyrus, temporal regions, and the visual word form area (VWFA). Further investigation of the activity in visual regions revealed overall deactivation relative to baseline rest for both attention conditions. Topographic analysis demonstrated that while attending to melody drove deactivation equivalently across all fusiform regions of interest examined, attending to speech produced a regionally specific modulation: deactivation of all fusiform regions, except the VWFA. Results indicate that selective attention to speech can topographically tune extrastriate cortex, leading to increased activity in VWFA relative to surrounding regions, in line with the well-established connectivity between areas related to spoken and visual word perception in skilled readers.

  14. Bactericidal activity of N-chlorotaurine against biofilm-forming bacteria grown on metal disks.

    PubMed

    Coraça-Huber, Débora C; Ammann, Christoph G; Fille, Manfred; Hausdorfer, Johann; Nogler, Michael; Nagl, Markus

    2014-01-01

    Many orthopedic surgeons consider surgical irrigation and debridement with prosthesis retention as a treatment option for postoperative infections. Usually, saline solution with no added antimicrobial agent is used for irrigation. We investigated the activity of N-chlorotaurine (NCT) against various biofilm-forming bacteria in vitro and thereby gained significant information on its usability as a soluble and well-tolerated active chlorine compound in orthopedic surgery. Biofilms of Staphylococcus aureus were grown on metal alloy disks and in polystyrene dishes for 48 h. Subsequently, they were incubated for 15 min to 7 h in buffered solutions containing therapeutically applicable concentrations of NCT (1%, 0.5%, and 0.1%; 5.5 to 55 mM) at 37°C. NCT inactivated the biofilm in a time- and dose-dependent manner. Scanning electron microscopy revealed disturbance of the biofilm architecture by rupture of the extracellular matrix. Assays with reduction of carboxanilide (XTT) showed inhibition of the metabolism of the bacteria in biofilms. Quantitative cultures confirmed killing of S. aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa biofilms on metal alloy disks by NCT. Clinical isolates were slightly more resistant than ATCC type strains, but counts of CFU were reduced at least 10-fold by 1% NCT within 15 min in all cases. NCT showed microbicidal activity against various bacterial strains in biofilms. Whether this can be transferred to the clinical situation should be the aim of future studies.

  15. 76 FR 34739 - Agency Information Collection Activities: HRIFA Instructions for Form I-485, Supplement C...

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  16. 77 FR 65703 - Agency Information Collection Activities: Application for Family Unity Benefits, Form I-817...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-30

    ... for Family Unity Benefits, Form I-817, Revision of a Currently Approved Collection ACTION: 60-Day...) Title of the Form/Collection: Application for Family Unity Benefits. (3) Agency form number, if any,...

  17. ESR dating of barite in sulphide deposits formed by the sea-floor hydrothermal activities.

    PubMed

    Toyoda, Shin; Fujiwara, Taisei; Uchida, Ai; Ishibashi, Jun-ichiro; Nakai, Shun'ichi; Takamasa, Asako

    2014-06-01

    Barite is a mineral newly found to be practically useful for electron spin resonance (ESR) dating of sulphide deposits formed by the sea-floor hydrothermal activities. The recent studies for the properties of the ESR dating signal in barite are summarised in the present paper as well as the formulas for corrections for accurate dose-rate estimation are developed including the dose-rate conversion factors, shape correction for gamma-ray dose and decay of (226)Ra. Although development of the techniques for ESR dating of barite has been completed, further comparative studies with other dating techniques such as U-Th and (226)Ra-(210)Pb dating are necessary for the technique to be widely used.

  18. Identification of Triclosan-O-Sulfate and other transformation products of Triclosan formed by activated sludge.

    PubMed

    Chen, Xijuan; Casas, Mònica Escolà; Nielsen, Jeppe Lund; Wimmer, Reinhard; Bester, Kai

    2015-02-01

    Aerobic degradation experiments of Triclosan were performed in activated sludge to identify possible transformation products for this compound. During 7 days, the formation of biotransformation products such as 2,4-Dichlorophenol, 4-Chlorocatechol, 5-Hydroxy-Triclosan and other Monohydroxy-Triclosan derivatives as well as Dihydroxy-Triclosan-derivatives were observed. The structure of 5-Hydroxy-Triclosan was elucidated by NMR data for the first time in sludge degradation experiments. Additionally the production of a hitherto unknown transformation product in sludge, i.e., Triclosan-O-Sulfate was detected. During the incubations, the concentrations of this transformation product changed from zero to 330 μg L(-1). Based on the analysis of the biodegradation products, three types of reactions were identified: 1) chemical scission of ether bond to form phenols and catechols, 2) addition of OH moieties to the aromatic ring, and 3) adding of methyl or sulfate groups to the original hydroxyl group.

  19. [Sports and recreational activities as a form of functional treatment of patients with ankylosing spondilytis].

    PubMed

    Grubisić, Frane; Grazio, Simeon; Znika, Matea

    2007-01-01

    Ankylosing spondylitis is a chronic inflammatory rheumatic disease that primarily affects the sacroiliac joints and spine, although it may involve entheses, peripheral joints and extraarticular organs. Disease treatment is directed toward the suppression of the inflammatory process and the improvement of the musculoskeletal system function. There are several treatment modalities: education of the patient and members of the family, pharmacological treatment, physical therapy and, in some cases, surgical treatment. An important segment of various modalities of physical therapy belongs to kinesitherapy, sports and recreation whose duration and intensity largely depends not only on the actual functional impairments, but also on the presence of some other disease or contraindications. Kinesitherapy is directed toward maintenance and improvement of the function of the spine, thoracic cavity and large synovial joints as well as the prevention of deformities or contractures. Kinesitherapy and sports programmes may involve individual or group approach. Patients are encouraged to participate in sports activities that may imitate or substitute specific forms of exercises.

  20. The role of biofilms in the sedimentology of actively forming gypsum deposits at Guerrero Negro, Mexico.

    PubMed

    Vogel, Marilyn B; Des Marais, David J; Turk, Kendra A; Parenteau, Mary N; Jahnke, Linda L; Kubo, Michael D Y

    2009-11-01

    Actively forming gypsum deposits at the Guerrero Negro sabkha and saltern system provided habitats for stratified, pigmented microbial communities that exhibited significant morphological and phylogenetic diversity. These deposits ranged from meter-thick gypsum crusts forming in saltern seawater concentration ponds to columnar microbial mats with internally crystallized gypsum granules developing in natural anchialine pools. Gypsum-depositing environments were categorized as forming precipitation surfaces, biofilm-supported surfaces, and clastic surfaces. Each surface type was described in terms of depositional environment, microbial diversity, mineralogy, and sedimentary fabrics. Precipitation surfaces developed in high-salinity subaqueous environments where rates of precipitation outpaced the accumulation of clastic, organic, and/or biofilm layers. These surfaces hosted endolithic biofilms comprised predominantly of oxygenic and anoxygenic phototrophs, sulfate-reducing bacteria, and bacteria from the phylum Bacteroidetes. Biofilm-supported deposits developed in lower-salinity subaqueous environments where light and low water-column turbulence supported dense benthic microbial communities comprised mainly of oxygenic phototrophs. In these settings, gypsum granules precipitated in the extracellular polymeric substance (EPS) matrix as individual granules exhibiting distinctive morphologies. Clastic surfaces developed in sabkha mudflats that included gypsum, carbonate, and siliclastic particles with thin gypsum/biofilm components. Clastic surfaces were influenced by subsurface brine sheets and capillary evaporation and precipitated subsedimentary gypsum discs in deeper regions. Biofilms appeared to influence both chemical and physical sedimentary processes in the various subaqueous and subaerially exposed environments studied. Biofilm interaction with chemical sedimentary processes included dissolution and granularization of precipitation surfaces, formation of

  1. Characteristics of dissolved organic matter formed in aerobic and anaerobic digestion of excess activated sludge.

    PubMed

    Du, Haixia; Li, Fusheng

    2017-02-01

    The characteristics of dissolved organic matter (DOM) formed in aerobic and anaerobic digestion of excess activated sludge (EAS) was investigated for three total solid (TS) concentrations (1.2, 2.3 and 5.2%) and three temperatures (5, 20 and 35 °C). The results on the overall concentration of DOM evaluated by TOC showed significantly higher values in anaerobic than aerobic digestion (2.8-6.9 times for TS 1.2-5.2% at 20 °C). Data analysis with a first-order sequential reaction model revealed that higher occurrence of DOM in anaerobic digestion was a result of comparatively faster hydrolysis (1.3-5.5 times for TS 1.2-5.2% at 20 °C; 1.4-49.3 times for temperatures 5-35 °C with TS 1.2%) and slower degradation (0.3-1.0 times for TS 1.2-5.2% at 20 °C; 0.5-8.3 times for temperatures 5-35 °C with TS 1.2%). In aerobic digestion, more humic substances were formed; while, in anaerobic digestion, proteins and aromatic amino acids were the major constituents. For both digestions, except for a few exceptions, proteins and humic substances increased as the TS concentration increased; and increasing the temperature led to a decrease in the content of proteins formed in both aerobic and anaerobic digestion, and an increase in the content of humic substances in the aerobic digestion. The UV-absorbing DOM constituents were highly heterogeneous, and were comparatively larger in anaerobic digestion; and did not change significantly with the TS concentrations and temperatures.

  2. Active monomeric and dimeric forms of Pseudomonas putida glyoxalase I: evidence for 3D domain swapping.

    PubMed

    Saint-Jean, A P; Phillips, K R; Creighton, D J; Stone, M J

    1998-07-21

    3D domain swapping of proteins involves the interconversion of a monomer containing a single domain-domain interface and a 2-fold symmetrical dimer containing two equivalent intermolecular interfaces. Human glyoxalase I has the structure of a domain-swapped dimer [Cameron, A. D., Olin, B., Ridderström, M., Mannervik, B., and Jones, T. A. (1997) EMBO J. 16, 3386-3395] but Pseudomonas putida glyoxalase I has been reported to be monomeric [Rhee, H.-I., Murata, K., and Kimura, A. (1986) Biochem. Biophys. Res. Commun. 141, 993-999]. We show here that recombinant P. putida glyoxalase I is an active dimer (kcat approximately 500 +/- 100 s-1; KM approximately 0.4 +/- 0.2 mM) with two zinc ions per dimer. The zinc is required for structure and function. However, treatment of the dimer with glutathione yields an active monomer (kcat approximately 115 +/- 40 s-1; KM approximately 1.4 +/- 0.4 mM) containing a single zinc ion. The monomer is metastable and slowly reverts to the active dimer in the absence of glutathione. Thus, glyoxalase I appears to be a novel example of a single protein able to exist in two alternative domain-swapped forms. It is unique among domain-swapped proteins in that the active site and an essential metal binding site are apparently disassembled and reassembled by the process of domain swapping. Furthermore, it is the only example to date in which 3D domain swapping can be regulated by a small organic ligand.

  3. Obscured active galactic nuclei triggered in compact star-forming galaxies

    NASA Astrophysics Data System (ADS)

    Chang, Yu-Yen; Le Floc'h, Emeric; Juneau, Stéphanie; da Cunha, Elisabete; Salvato, Mara; Civano, Francesca; Marchesi, Stefano; Gabor, J. M.; Ilbert, Olivier; Laigle, Clotilde; McCracken, H. J.; Hsieh, Bau-Ching; Capak, Peter

    2017-03-01

    We present a structural study of 182 obscured active galactic nuclei (AGNs) at z ≤ 1.5, selected in the Cosmic Evolution Survey field from their extreme infrared to X-ray luminosity ratio and their negligible emission at optical wavelengths. We fit optical to far-infrared spectral energy distributions and analyse deep Hubble Space Telescope imaging to derive the physical and morphological properties of their host galaxies. We find that such galaxies are more compact than normal star-forming sources at similar redshift and stellar mass, and we show that it is not an observational bias related to the emission of the AGN. Based on the distribution of their UVJ colours, we also argue that this increased compactness is not due to the additional contribution of a passive bulge. We thus postulate that a vast majority of obscured AGNs reside in galaxies undergoing dynamical compaction, similar to processes recently invoked to explain the formation of compact star-forming sources at high redshift.

  4. Cooperation of Doxycycline with Phytochemicals and Micronutrients Against Active and Persistent Forms of Borrelia sp.

    PubMed

    Goc, Anna; Niedzwiecki, Alexandra; Rath, Matthias

    2016-01-01

    Phytochemicals and micronutrients represent a growing theme in antimicrobial defense; however, little is known about their anti-borreliae effects of reciprocal cooperation with antibiotics. A better understanding of this aspect could advance our knowledge and help improve the efficacy of current approaches towards Borrelia sp. In this study, phytochemicals and micronutrients such as baicalein, luteolin, 10-HAD, iodine, rosmarinic acid, and monolaurin, as well as, vitamins D3 and C were tested in a combinations with doxycycline for their in vitro effectiveness against vegetative (spirochetes) and latent (rounded bodies, biofilm) forms of Borrelia burgdorferi and Borrelia garinii. Anti-borreliae effects were evaluated according to checkerboard assays and supported by statistical analysis. The results showed that combination of doxycycline with flavones such as baicalein and luteolin exhibited additive effects against all morphological forms of studied Borrelia sp. Doxycycline combined with iodine demonstrated additive effects against spirochetes and biofilm, whereas with fatty acids such as monolaurin and 10-HAD it produced FICIs of indifference. Additive anti-spirochetal effects were also observed when doxycycline was used with rosmarinic acid and both vitamins D3 and C. Antagonism was not observed in any of the cases. This data revealed the intrinsic anti-borreliae activity of doxycycline with tested phytochemicals and micronutrients indicating that their addition may enhance efficacy of this antibiotic in combating Borrelia sp. Especially the addition of flavones balcalein and luteolin to a doxycycline regimen could be explored further in defining more effective treatments against these bacteria.

  5. Isozymic forms of rat brain CA/sup 2 +/-activated and phospholipid-dependent protein kinase

    SciTech Connect

    Huang, K.P.; Huang, F.L.

    1986-05-01

    Three forms of Ca/sup 2 +/-activated and phospholipid-dependent protein kinase (protein kinase C) were purified from the cytosolic fraction of rat brain. These enzymes, designated as type I, II, and III protein kinase C, all have the similar molecular weight of 80 Kd, bind (/sup 3/H)-phorbol dibutyrate in the presence of Ca/sup 2 +/, and undergo autophosphorylation in the presence of Ca/sup 2 +/, phosphatidylserine, and diolein. Autophosphorylation of these kinases resulted in an incorporation of 1- 1.5 mol /sup 32/P/mol of enzyme. Analysis of the /sup 32/P-labeled tryptic peptides derived from the autophosphorylated protein kinase C by two-dimensional peptide mapping revealed that these kinases had different sites of autophosphorylation. Phosphoamino acid analysis revealed that the type I and type III protein kinase C mainly phosphorylated at Ser residue while the type II kinase phosphorylated at both Ser and Thr residues. In addition, polyclonal antibodies previously prepared against a mixed enzyme fraction preferentially inhibited the type I and type II enzymes but less effectively toward the type III enzyme. Monoclonal antibody specifically against the type II protein kinase C did not inhibit the type I or type III enzymes. These kinases also had different susceptibility to limited proteolysis by trypsin and upon proteolytic degradation they generate distinct fragments. These results demonstrate the presence of isozymic forms of protein kinase C in rat brain.

  6. Cooperation of Doxycycline with Phytochemicals and Micronutrients Against Active and Persistent Forms of Borrelia sp

    PubMed Central

    Goc, Anna; Niedzwiecki, Alexandra; Rath, Matthias

    2016-01-01

    Phytochemicals and micronutrients represent a growing theme in antimicrobial defense; however, little is known about their anti-borreliae effects of reciprocal cooperation with antibiotics. A better understanding of this aspect could advance our knowledge and help improve the efficacy of current approaches towards Borrelia sp. In this study, phytochemicals and micronutrients such as baicalein, luteolin, 10-HAD, iodine, rosmarinic acid, and monolaurin, as well as, vitamins D3 and C were tested in a combinations with doxycycline for their in vitro effectiveness against vegetative (spirochetes) and latent (rounded bodies, biofilm) forms of Borrelia burgdorferi and Borrelia garinii. Anti-borreliae effects were evaluated according to checkerboard assays and supported by statistical analysis. The results showed that combination of doxycycline with flavones such as baicalein and luteolin exhibited additive effects against all morphological forms of studied Borrelia sp. Doxycycline combined with iodine demonstrated additive effects against spirochetes and biofilm, whereas with fatty acids such as monolaurin and 10-HAD it produced FICIs of indifference. Additive anti-spirochetal effects were also observed when doxycycline was used with rosmarinic acid and both vitamins D3 and C. Antagonism was not observed in any of the cases. This data revealed the intrinsic anti-borreliae activity of doxycycline with tested phytochemicals and micronutrients indicating that their addition may enhance efficacy of this antibiotic in combating Borrelia sp. Especially the addition of flavones balcalein and luteolin to a doxycycline regimen could be explored further in defining more effective treatments against these bacteria. PMID:27570483

  7. Microwave-assisted extraction of active pharmaceutical ingredient from solid dosage forms.

    PubMed

    Hoang, T H; Sharma, R; Susanto, D; Di Maso, M; Kwong, E

    2007-07-13

    The microwave assisted extraction (MAE) technique has been evaluated for the extraction of active pharmaceutical ingredients (API) from various solid dosage forms. Using immediate release tablets of Compound A as a model, optimization of the extraction method with regards to extraction solvent composition, extraction time and temperature was briefly discussed. Complete recovery of Compound A was achieved when samples were extracted using acetonitrile as the extraction solvent under microwave heating at a constant cell temperature of 50 degrees C for 5 min. The optimized MAE method was applied for content uniformity (single tablet extraction) and potency (multiple tablets extraction) assays of release and stability samples of two products of Compound A (5 and 25mg dose strength) stored at various conditions. To further demonstrate the applicability of MAE, the instrumental extraction conditions (50 degrees C for 5 min) were adopted for the extraction of montelukast sodium (Singulair) from various solid dosage forms using methanol-water (75:25, v/v) as the extraction solvent. The MAE procedure demonstrated an extraction efficiency of 97.4-101.9% label claim with the greatest RSD at 1.4%. The results compare favorably with 97.6-102.3% label claim with the greatest RSD at 2.9% obtained with validated mechanical extraction procedures. The system is affordable, user-friendly and simple to operate and troubleshoot. Rapid extraction process (7 min/run) along with high throughput capacity (up to 23 samples simultaneously) would lead to reduced cycle time and thus increased productivity.

  8. Activation of human natural killer cells by the soluble form of cellular prion protein.

    PubMed

    Seong, Yeon-Jae; Sung, Pil Soo; Jang, Young-Soon; Choi, Young Joon; Park, Bum-Chan; Park, Su-Hyung; Park, Young Woo; Shin, Eui-Cheol

    2015-08-21

    Cellular prion protein (PrP(C)) is widely expressed in various cell types, including cells of the immune system. However, the specific roles of PrP(C) in the immune system have not been clearly elucidated. In the present study, we investigated the effects of a soluble form of recombinant PrP(C) protein on human natural killer (NK) cells. Recombinant soluble PrP(C) protein was generated by fusion of human PrP(C) with the Fc portion of human IgG1 (PrP(C)-Fc). PrP(C)-Fc binds to the surface of human NK cells, particularly to CD56(dim) NK cells. PrP(C)-Fc induced the production of cytokines and chemokines and the degranulation of granzyme B from NK cells. In addition, PrP(C)-Fc facilitated the IL-15-induced proliferation of NK cells. PrP(C)-Fc induced phosphorylation of ERK-1/2 and JNK in NK cells, and inhibitors of the ERK or the JNK pathways abrogated PrP(C)-Fc-induced cytokine production in NK cells. In conclusion, the soluble form of recombinant PrP(C)-Fc protein activates human NK cells via the ERK and JNK signaling pathways.

  9. Parallel or serial activation of word forms in speech production? Neurolinguistic evidence from an aphasic patient.

    PubMed

    Blanken, Gerhard; Dittmann, Jürgen; Wallesch, Claus-W

    2002-05-31

    We report the oral picture naming performance of the German aphasic MW who presented with frequent meaning related word substitutions (e.g. tiger ==> lion) and word finding blockings (omissions) while his phonological capacities at the single word level were nearly preserved. Targets were controlled for their 'semantic competitiveness', that is, whether there exist closely meaning related lexical competitors or not. Semantic errors were far more numerous with the highly competitive targets than with the low competitive ones. However, omissions were more frequent with the low competitive items so that the sum of the semantic errors and of the omissions was comparable in both conditions. This inverse and compensatory relationship suggests that both error types are not mutually independent. The found pattern is at odds with serial psycholinguistic theories which locate word selection (and misselection) and word form access (and blockings) at different and serially connected stages of word production but supports theories which allow for a parallel architecture in lexical activation and selection involving the word form level.

  10. Activation of human natural killer cells by the soluble form of cellular prion protein

    SciTech Connect

    Seong, Yeon-Jae; Sung, Pil Soo; Jang, Young-Soon; Choi, Young Joon; Park, Bum-Chan; Park, Su-Hyung; Park, Young Woo; Shin, Eui-Cheol

    2015-08-21

    Cellular prion protein (PrP{sup C}) is widely expressed in various cell types, including cells of the immune system. However, the specific roles of PrP{sup C} in the immune system have not been clearly elucidated. In the present study, we investigated the effects of a soluble form of recombinant PrP{sup C} protein on human natural killer (NK) cells. Recombinant soluble PrP{sup C} protein was generated by fusion of human PrP{sup C} with the Fc portion of human IgG{sub 1} (PrP{sup C}-Fc). PrP{sup C}-Fc binds to the surface of human NK cells, particularly to CD56{sup dim} NK cells. PrP{sup C}-Fc induced the production of cytokines and chemokines and the degranulation of granzyme B from NK cells. In addition, PrP{sup C}-Fc facilitated the IL-15-induced proliferation of NK cells. PrP{sup C}-Fc induced phosphorylation of ERK-1/2 and JNK in NK cells, and inhibitors of the ERK or the JNK pathways abrogated PrP{sup C}-Fc-induced cytokine production in NK cells. In conclusion, the soluble form of recombinant PrP{sup C}-Fc protein activates human NK cells via the ERK and JNK signaling pathways. - Highlights: • Recombinant soluble PrP{sup C} (PrP{sup C}-Fc) was generated by fusion of human PrP{sup C} with IgG1 Fc portion. • PrP{sup C}-Fc protein induces the production of cytokines and degranulation from human NK cells. • PrP{sup C}-Fc protein enhances the IL-15-induced proliferation of human NK cells. • PrP{sup C}-Fc protein activates human NK cells via the ERK and JNK signaling pathways.

  11. CTEPP DATA COLLECTION FORM 08 (PERIODS 1-5 AND FOOD, FRUIT & VEG): CHILD ACTIVITY DIARY AND FOOD SURVEY

    EPA Science Inventory

    This data collection form is divided into two parts: Child Activity Diary and Food Survey. The Child Activity Diary collects information on the child's activities at home over the 48-hr monitoring period. The diary is divided into five time periods over the 48-hr monitoring inter...

  12. CTEPP DATA COLLECTION FORM 09 (PERIODS 1-4 AND FOOD, FRUIT & VEG): CHILD ACTIVITY DIARY AND FOOD SURVEY

    EPA Science Inventory

    This data collection form is divided into two parts: Child Activity Diary and Food Survey. The Child Activity Diary collects information on the child's activities at home over the 48-hr monitoring period. The diary is divided into four time periods over the 48-hr monitoring inter...

  13. Antibody nanoparticle dispersions formed with mixtures of crowding molecules retain activity and in vivo bioavailability.

    PubMed

    Miller, Maria A; Khan, Tarik A; Kaczorowski, Kevin J; Wilson, Brian K; Dinin, Aileen K; Borwankar, Ameya U; Rodrigues, Miguel A; Truskett, Thomas M; Johnston, Keith P; Maynard, Jennifer A

    2012-10-01

    Monoclonal antibodies continue to command a large market for treatment of a variety of diseases. In many cases, the doses required for therapeutic efficacy are large, limiting options for antibody delivery and administration. We report a novel formulation strategy based on dispersions of antibody nanoclusters that allows for subcutaneous injection of highly concentrated antibody (≈ 190 mg/mL). A solution of monoclonal antibody 1B7 was rapidly frozen and lyophilized using a novel spiral-wound in-situ freezing technology to generate amorphous particles. Upon gentle stirring, a translucent dispersion of approximately 430 nm protein clusters with low apparent viscosity (≈ 24 cp) formed rapidly in buffer containing the pharmaceutically acceptable crowding agents such as trehalose, polyethylene glycol, and n-methyl-2-pyrrolidone. Upon in vitro dilution of the dispersion, the nanoclusters rapidly reverted to monomeric protein with full activity, as monitored by dynamic light scattering and antigen binding. When administered to mice as an intravenous solution, subcutaneous solution, or subcutaneous dispersion at similar (4.6-7.3 mg/kg) or ultra-high dosages (51.6 mg/kg), the distribution and elimination kinetics were within error and the protein retained full activity. Overall, this method of generating high-concentration, low-viscosity dispersions of antibody nanoclusters could lead to improved administration and patient compliance, providing new opportunities for the biotechnology industry.

  14. Pore-forming activity of OmpA protein of Escherichia coli.

    PubMed

    Sugawara, E; Nikaido, H

    1992-02-05

    Escherichia coli outer membrane protein OmpA was purified to homogeneity, as a monomer, from a K12 derivative deficient in both OmpF and OmpC porins. When proteoliposomes reconstituted from the purified OmpA, phospholipids, and lithium dodecyl sulfate were tested for permeability to small molecules by osmotic swelling, it was found that OmpA produced apparently nonspecific diffusion channels that allowed the penetration of various solutes. The pore-forming activity was destroyed by the heat denaturation of the OmpA protein, and the use of an OmpA-deficient mutant showed that the activity was not caused by copurifying contaminants. The size of the OmpA channel, estimated by comparison of diffusion rates of solutes of different sizes, was rather similar to that of E. coli OmpF and OmpC porins, i.e. about 1 nm in diameter. The rate of penetration of L-arabinose caused by a given amount of OmpA protein, however, was about a hundredfold lower than the rate produced by the same amount of E. coli OmpF porin. The addition of large amounts of lithium dodecyl sulfate to the reconstitution mixture increased the permeability through the OmpA channel, apparently by facilitating the correct insertion of OmpA into the bilayer.

  15. Alarin but not its alternative-splicing form, GALP (Galanin-like peptide) has antimicrobial activity

    SciTech Connect

    Wada, Akihiro; Wong, Pooi-Fong; Hojo, Hironobu; Hasegawa, Makoto; Ichinose, Akitoyo; Llanes, Rafael; Kubo, Yoshinao; Senba, Masachika; Ichinose, Yoshio

    2013-05-03

    Highlights: • Alarin inhibits the growth of E. coli but not S. aureus. • Alarin’s potency is comparable to LL-37 in inhibiting the growth of E. coli. • Alarin can cause bacterial membrane blebbing. • Alalin does not induce hemolysis on erythrocytes. -- Abstract: Alarin is an alternative-splicing form of GALP (galanin-like peptide). It shares only 5 conserved amino acids at the N-terminal region with GALP which is involved in a diverse range of normal brain functions. This study seeks to investigate whether alarin has additional functions due to its differences from GALP. Here, we have shown using a radial diffusion assay that alarin but not GALP inhibited the growth of Escherichia coli (strain ML-35). The conserved N-terminal region, however, remained essential for the antimicrobial activity of alarin as truncated peptides showed reduced killing effect. Moreover, alarin inhibited the growth of E. coli in a similar potency as human cathelicidin LL-37, a well-studied antimicrobial peptide. Electron microscopy further showed that alarin induced bacterial membrane blebbing but unlike LL-37, it did not cause hemolysis of erythrocytes. In addition, alarin is only active against the gram-negative bacteria, E. coli but not the gram-positive bacteria, Staphylococcus aureus. Thus, these data suggest that alarin has potentials as an antimicrobial and should be considered for the development in human therapeutics.

  16. WIDESPREAD AND HIDDEN ACTIVE GALACTIC NUCLEI IN STAR-FORMING GALAXIES AT REDSHIFT >0.3

    SciTech Connect

    Juneau, Stephanie; Bournaud, Frederic; Daddi, Emanuele; Elbaz, David; Alexander, David M.; Mullaney, James R.; Magnelli, Benjamin; Hwang, Ho Seong; Willner, S. P.; Coil, Alison L.; Rosario, David J.; Trump, Jonathan R.; Faber, S. M.; Kocevski, Dale D.; Cooper, Michael C.; Frayer, David T.; and others

    2013-02-20

    We characterize the incidence of active galactic nuclei (AGNs) in 0.3 < z < 1 star-forming galaxies by applying multi-wavelength AGN diagnostics (X-ray, optical, mid-infrared, radio) to a sample of galaxies selected at 70 {mu}m from the Far-Infrared Deep Extragalactic Legacy survey (FIDEL). Given the depth of FIDEL, we detect 'normal' galaxies on the specific star formation rate (sSFR) sequence as well as starbursting systems with elevated sSFR. We find an overall high occurrence of AGN of 37% {+-} 3%, more than twice as high as in previous studies of galaxies with comparable infrared luminosities and redshifts but in good agreement with the AGN fraction of nearby (0.05 < z < 0.1) galaxies of similar infrared luminosities. The more complete census of AGNs comes from using the recently developed Mass-Excitation (MEx) diagnostic diagram. This optical diagnostic is also sensitive to X-ray weak AGNs and X-ray absorbed AGNs, and reveals that absorbed active nuclei reside almost exclusively in infrared-luminous hosts. The fraction of galaxies hosting an AGN appears to be independent of sSFR and remains elevated both on the sSFR sequence and above. In contrast, the fraction of AGNs that are X-ray absorbed increases substantially with increasing sSFR, possibly due to an increased gas fraction and/or gas density in the host galaxies.

  17. Photoluminescence Mechanism and Photocatalytic Activity of Organic-Inorganic Hybrid Materials Formed by Sequential Vapor Infiltration.

    PubMed

    Akyildiz, Halil I; Stano, Kelly L; Roberts, Adam T; Everitt, Henry O; Jur, Jesse S

    2016-05-03

    Organic-inorganic hybrid materials formed by sequential vapor infiltration (SVI) of trimethylaluminum into polyester fibers are demonstrated, and the photoluminescence of the fibers is evaluated using a combined UV-vis and photoluminescence excitation (PLE) spectroscopy approach. The optical activity of the modified fibers depends on infiltration thermal processing conditions and is attributed to the reaction mechanisms taking place at different temperatures. At low temperatures a single excitation band and dual emission bands are observed, while, at high temperatures, two distinct absorption bands and one emission band are observed, suggesting that the physical and chemical structure of the resulting hybrid material depends on the SVI temperature. Along with enhancing the photoluminescence intensity of the PET fibers, the internal quantum efficiency also increased to 5-fold from ∼4-5% to ∼24%. SVI processing also improved the photocatalytic activity of the fibers, as demonstrated by photodeposition of Ag and Au metal particles out of an aqueous metal salt solution onto fiber surfaces via UVA light exposure. Toward applications in flexible electronics, well-defined patterning of the metallic materials is achieved by using light masking and focused laser rastering approaches.

  18. Antibody nanoparticle dispersions formed with mixtures of crowding molecules retain activity and in vivo bioavailability

    PubMed Central

    Miller, Maria A.; Khan, Tarik A.; Kaczorowski, Kevin J.; Wilson, Brian K.; Dinin, Aileen K.; Borwankar, Ameya U.; Rodrigues, Miguel A.; Truskett, Thomas M.; Johnston, Keith P.; Maynard, Jennifer A.

    2013-01-01

    Monoclonal antibodies continue to command a large market for treatment of a variety of diseases. In many cases, the doses required for therapeutic efficacy are large, limiting options for antibody delivery and administration. We report a novel formulation strategy based on dispersions of antibody nanoclusters that allows for subcutaneous injection of highly concentrated antibody (~190 mg/ml). A solution of monoclonal antibody 1B7 was rapidly frozen and lyophilized using a novel spiral-wound in situ freezing technology (SWIFT) to generate amorphous particles. Upon gentle stirring, a translucent dispersion of ~430 nm protein clusters low apparent viscosity (~24 cp) formed rapidly in buffer containing the pharmaceutically acceptable crowding agents, trehalose, polyethylene glycol and n-methyl-2-pyrrolidone. Upon in vitro dilution of the dispersion, the nanoclusters rapidly reverted to monomeric protein with full activity, as monitored by dynamic light scattering and antigen binding. When administered to mice as an intravenous solution, subcutaneous solution or subcutaneous dispersion at similar (4.6-7.3 mg/kg) or ultra-high dosages (51.6 mg/kg), the distribution and elimination kinetics were within error and the protein retained full activity. Overall, this method of generating high-concentration, low-viscosity dispersions of antibody nanoclusters could lead to improved administration and patient compliance, providing new opportunities for the biotechnology industry. PMID:22777686

  19. Locally formed dopamine inhibits Na sup + -K sup + -ATPase activity in rat renal cortical tubule cells

    SciTech Connect

    Seri, I.; Kone, B.C.; Gullans, S.R.; Aperia, A.; Brenner, B.M.; Ballermann, B.J. Karolinska Institute, Stockholm )

    1988-10-01

    Dopamine, generated locally from L-dopa, inhibits Na{sup +}-K{sup +}-ATPase in permeabilized rat proximal tubules under maximum transport rate conditions for sodium. To determine whether locally formed dopamine inhibits Na{sup +}-K{sup +}-ATPase activity in intact cortical tubule cells we studied the effect of L-dopa on ouabain-sensitive oxygen consumption rate ({dot Q}o{sub 2}) and {sup 86}Rb uptake in renal cortical tubule cell suspensions. L-Dopa did not affect ouabain-insensitive {dot Q}o{sub 2} or mitochondrial respiration. However, L-dopa inhibited ouabain-sensitive {dot Q}o{sub 2} in a concentration-dependent manner, with half-maximal inhibition (K{sub 0.5}) of 5 {times} 10{sup {minus}7} M and a maximal inhibition of 14.1 {plus minus} 1.5% at 10{sup {minus}4}M. L-Dopa also blunted the nystatin-stimulated {dot Q}o{sub 2} in a concentration-dependent manner, indicating the L-dopa directly inhibits Na{sup +}-K{sup +}-ATPase activity and not sodium entry. Ouabain-sensitive {sup 86}Rb uptake was also inhibited by L-dopa. Carbidopa, an inhibitor of the conversion of L-dopa to dopamine, eliminated the effect of L-dopa on ouabain-sensitive {dot Q}o{sub 2} and {sup 86}Rb uptake, indicating that dopamine rather than L-dopa was the active agent. The finding that the L-dopa concentration-response curve was shifted to the left by one order of magnitude in the presence of nystatin suggests that the inhibitory effect is enhanced when the intracellular sodium concentration is increased. By studying the effect of L-dopa on ouabain-sensitive {dot Q}o{sub 2} at increasing extracellular sodium concentrations in the presence of nystatin, the authors demonstrated that the inhibitory effect of locally formed dopamine on the Na{sup +}-K{sup +}-ATPase is indeed dependent on the sodium available for the enzyme and occurs in an uncompetitive manner.

  20. 78 FR 27965 - Agency Information Collection Activities: Submission for OMB Review; Comment Request Re Forms...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-13

    ... Defined Contribution Plan, Form 7200/10 Declaration for IRA/KEOGH Deposit, Form 7200/11 Declaration for... Declaration for IRA/KEOGH Deposit, Form 7200/11 0.50 5 2.5 Declaration for Defined Benefit Plan, Form...

  1. 78 FR 73875 - Agency Information Collection Activities: Canadian Border Boat Landing Permit (CBP Form I-68)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-09

    ... Landing Permit (CBP Form I-68) AGENCY: U.S. Customs and Border Protection (CBP), Department of Homeland... requirement concerning the Canadian Border Boat Landing Permit (Form I-68). This request for comment is being... Form I-68. Abstract: The Canadian Border Boat Landing Permit (CBP Form I-68) allows...

  2. 76 FR 2917 - Agency Information Collection Activities: Canadian Border Boat Landing Permit (CBP Form I-68)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-18

    ... Landing Permit (CBP Form I-68) AGENCY: U.S. Customs and Border Protection, Department of Homeland Security... (CBP Form I-68). This is a proposed extension of an information collection that was previously approved... Landing Permit. OMB Number: 1651-0108. Form Number: CBP Form I-68. Abstract: The Canadian Border...

  3. 75 FR 61508 - Agency Information Collection Activities: Canadian Border Boat Landing Permit (CBP Form I-68)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-05

    ... Landing Permit (CBP Form I-68) AGENCY: U.S. Customs and Border Protection (CBP), Department of Homeland... concerning the Canadian Border Boat Landing Permit (Form I- ] 68). This request for comment is being made... Form I-68. Abstract: The Canadian Border Boat Landing Permit (CBP Form I-68) allows...

  4. 76 FR 72967 - Agency Information Collection Activities: Form I-602; Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-28

    ... Collection Under Review; Form I- 602, Application by Refugee for Waiver of Grounds of Excludability; OMB... the Form/Collection: Application by Refugee for Waiver of Grounds of Excludability. (3) Agency form... Application by Refugee for Waiver of Grounds of Excludability, Form I- 602, is necessary to...

  5. 77 FR 65706 - Agency Information Collection Activities: Immigrant Petition for Alien Worker, Form I-140...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-30

    ... Petition for Alien Worker, Form I-140; Revision of a Currently Approved Collection ACTION: 60-Day Notice... Form/Collection: Immigrant Petition for Alien Worker. (3) Agency form number, if any, and the... information furnished on Form I-140 will be used by USCIS to classify aliens under sections 203(b)(1),...

  6. Phospholipase A2-activating protein is associated with a novel form of leukoencephalopathy.

    PubMed

    Falik Zaccai, Tzipora C; Savitzki, David; Zivony-Elboum, Yifat; Vilboux, Thierry; Fitts, Eric C; Shoval, Yishay; Kalfon, Limor; Samra, Nadra; Keren, Zohar; Gross, Bella; Chasnyk, Natalia; Straussberg, Rachel; Mullikin, James C; Teer, Jamie K; Geiger, Dan; Kornitzer, Daniel; Bitterman-Deutsch, Ora; Samson, Abraham O; Wakamiya, Maki; Peterson, Johnny W; Kirtley, Michelle L; Pinchuk, Iryna V; Baze, Wallace B; Gahl, William A; Kleta, Robert; Anikster, Yair; Chopra, Ashok K

    2017-02-01

    Leukoencephalopathies are a group of white matter disorders related to abnormal formation, maintenance, and turnover of myelin in the central nervous system. These disorders of the brain are categorized according to neuroradiological and pathophysiological criteria. Herein, we have identified a unique form of leukoencephalopathy in seven patients presenting at ages 2 to 4 months with progressive microcephaly, spastic quadriparesis, and global developmental delay. Clinical, metabolic, and imaging characterization of seven patients followed by homozygosity mapping and linkage analysis were performed. Next generation sequencing, bioinformatics, and segregation analyses followed, to determine a loss of function sequence variation in the phospholipase A2-activating protein encoding gene (PLAA). Expression and functional studies of the encoded protein were performed and included measurement of prostaglandin E2 and cytosolic phospholipase A2 activity in membrane fractions of fibroblasts derived from patients and healthy controls. Plaa-null mice were generated and prostaglandin E2 levels were measured in different tissues. The novel phenotype of our patients segregated with a homozygous loss-of-function sequence variant, causing the substitution of leucine at position 752 to phenylalanine, in PLAA, which causes disruption of the protein's ability to induce prostaglandin E2 and cytosolic phospholipase A2 synthesis in patients' fibroblasts. Plaa-null mice were perinatal lethal with reduced brain levels of prostaglandin E2 The non-functional phospholipase A2-activating protein and the associated neurological phenotype, reported herein for the first time, join other complex phospholipid defects that cause leukoencephalopathies in humans, emphasizing the importance of this axis in white matter development and maintenance.

  7. Round-robin testing of a reference glass for low-activity waste forms

    SciTech Connect

    Ebert, W. L.; Wolf, S. F.

    1999-12-06

    A round robin test program was conducted with a glass that was developed for use as a standard test material for acceptance testing of low-activity waste glasses made with Hanford tank wastes. The glass is referred to as the low-activity test reference material (LRM). The program was conducted to measure the interlaboratory reproducibility of composition analysis and durability test results. Participants were allowed to select the methods used to analyze the glass composition. The durability tests closely followed the Product Consistency Test (PCT) Method A, except that tests were conducted at both 40 and 90 C and that parallel tests with a reference glass were not required. Samples of LRM glass that had been crushed, sieved, and washed to remove fines were provided to participants for tests and analyses. The reproducibility of both the composition and PCT results compare favorably with the results of interlaboratory studies conducted with other glasses. From the perspective of reproducibility of analysis results, this glass is acceptable for use as a composition standard for nonradioactive components of low-activity waste forms present at >0.1 elemental mass % and as a test standard for PCTS at 40 and 90 C. For PCT with LRM glass, the expected test results at the 95% confidence level are as follows: (1) at 40 C: pH = 9.86 {+-} 0.96; [B] = 2.30 {+-} 1.25 mg/L; [Na] = 19.7 {+-} 7.3 mg/L; [Si] = 13.7 {+-} 4.2 mg/L; and (2) at 90 C: pH = 10.92 {+-} 0.43; [B] = 26.7 {+-} 7.2 mg/L; [Na] = 160 {+-} 13 mg/L; [Si] = 82.0 {+-} 12.7 mg/L. These ranges can be used to evaluate the accuracy of PCTS conducted at other laboratories.

  8. The pretranslocation ribosome is targeted by GTP-bound EF-G in partially activated form

    PubMed Central

    Hauryliuk, Vasili; Mitkevich, Vladimir A.; Eliseeva, Natalia A.; Petrushanko, Irina Yu.; Ehrenberg, Måns; Makarov, Alexander A.

    2008-01-01

    Translocation of the tRNA·mRNA complex through the bacterial ribosome is driven by the multidomain guanosine triphosphatase elongation factor G (EF-G). We have used isothermal titration calorimetry to characterize the binding of GDP and GTP to free EF-G at 4°C, 20°C, and 37°C. The binding affinity of EF-G is higher to GDP than to GTP at 4°C, but lower at 37°C. The binding enthalpy and entropy change little with temperature in the case of GDP binding but change greatly in the case of GTP binding. These observations are compatible with a large decrease in the solvent-accessible hydrophobic surface area of EF-G on GTP, but not GDP, binding. The explanation we propose is the locking of the switch 1 and switch 2 peptide loops in the G domain of EF-G to the γ-phosphate of GTP. From these data, in conjunction with previously reported structural data on guanine nucleotide-bound EF-G, we suggest that EF-G enters the pretranslocation ribosome as an “activity chimera,” with the G domain activated by the presence of GTP but the overall factor conformation in the inactive form typical of a GDP-bound multidomain guanosine triphosphatase. We propose that the active overall conformation of EF-G is attained only in complex with the ribosome in its “ratcheted state,” with hybrid tRNA binding sites. PMID:18836081

  9. Purified TMEM16A is sufficient to form Ca2+-activated Cl− channels

    PubMed Central

    Terashima, Hiroyuki; Picollo, Alessandra; Accardi, Alessio

    2013-01-01

    Ca2+-activated Cl− channels (CaCCs) are key regulators of numerous physiological functions, ranging from electrolyte secretion in airway epithelia to cellular excitability in sensory neurons and muscle fibers. Recently, TMEM16A (ANO1) and -B were shown to be critical components of CaCCs. It is still unknown whether they are also sufficient to form functional CaCCs, or whether association with other subunits is required. Recent reports suggest that the Ca2+ sensitivity of TMEM16A is mediated by its association with calmodulin, suggesting that functional CaCCs are heteromultimers. To test whether TMEM16A is necessary and sufficient to form functional CaCCs, we expressed, purified, and reconstituted human TMEM16A. The purified protein mediates Ca2+-dependent Cl− transport with submicromolar sensitivity to Ca2+, consistent with what is seen in patch–clamp experiments. The channel is synergistically gated by Ca2+ and voltage, so that opening is promoted by depolarizing potentials. Mutating two conserved glutamates in the TM6-7 intracellular loop selectively abolishes the Ca2+ dependence of reconstituted TMEM16A, in a manner similar to what was reported for the heterologously expressed channel. Well-characterized CaCC blockers inhibit Cl− transport with Kis comparable to those measured for native and heterologously expressed CaCCs. Finally, direct physical interactions between calmodulin and TMEM16A could not be detected in copurification experiments or in functional assays. Our results demonstrate that purified TMEM16A is necessary and sufficient to recapitulate the biophysical and pharmacological properties of native and heterologously expressed CaCCs. Our results also show that association of TMEM16A with other proteins, such as calmodulin, is not required for function. PMID:24167264

  10. Enhanced bone-forming activity of side population cells in the periodontal ligament.

    PubMed

    Ninomiya, Tadashi; Hiraga, Toru; Hosoya, Akihiro; Ohnuma, Kiyoshi; Ito, Yuzuru; Takahashi, Masafumi; Ito, Susumu; Asashima, Makoto; Nakamura, Hiroaki

    2014-04-01

    Regeneration of alveolar bone is critical for the successful treatment of periodontal diseases. The periodontal ligament (PDL) has been widely investigated as a source of cells for the regeneration of periodontal tissues. In the present study where we attempted to develop an effective strategy for alveolar bone regeneration, we examined the osteogenic potential of side population (SP) cells, a stem cell-containing population that has been shown to be highly abundant in several kinds of tissues, in PDL cells. Isolated SP cells from the rat PDL exhibited a superior ability to differentiate into osteoblastic cells compared with non-SP (NSP) and unsorted PDL cells in vitro. The mRNA expressions of osteoblast markers and bone morphogenetic protein (BMP) 2 were significantly upregulated in SP cells and were further increased by osteogenic induction. To examine the bone-forming activity of SP cells in vivo, PDL SP cells isolated from green fluorescent protein (GFP)-transgenic rats were transplanted with hydroxyapatite (HA) disks into wild-type animals. SP cells exhibited a high ability to induce the mineralized matrix compared with NSP and unsorted PDL cells. At 12 weeks after the implantation, some of the pores in the HA disks with SP cells were filled with mineralized matrices, which were positive for bone matrix proteins, such as osteopontin, bone sialoprotein, and osteocalcin. Furthermore, osteoblast- and osteocyte-like cells on and in the bone-like mineralized matrices were GFP positive, suggesting that the matrices were directly formed by the transplanted cells. These results suggest that PDL SP cells possess enhanced osteogenic potential and could be a potential source for cell-based regenerative therapy for alveolar bone.

  11. Residues involved in the pore-forming activity of the Clostridium perfringens iota toxin.

    PubMed

    Knapp, Oliver; Maier, Elke; Waltenberger, Eva; Mazuet, Christelle; Benz, Roland; Popoff, Michel R

    2015-02-01

    Clostridium perfringens iota toxin is a binary toxin that is organized into enzyme (Ia) and binding (Ib) components. Ib forms channels in lipid bilayers and mediates the transport of Ia into the target cells. Here we show that Ib residues 334-359 contain a conserved pattern of alternating hydrophobic and hydrophilic residues forming two amphipathic β-strands involved in membrane insertion and channel formation. This stretch of amino acids shows remarkable structural and functional analogies with the β-pore-forming domain of C. perfringens epsilon toxin. Several mutations within the two amphipathic β-strands affected pore formation, single-channel conductance and ion selectivity (S339E-S341E, Q345H N346E) confirming their involvement in channel formation. F454 of Ib corresponds to the Φ-clamp F427 of anthrax protective antigen and F428 of C2II binary toxins. The mutation F454A resulted in a loss of cytotoxicity and strong increase in single-channel conductance (500 pS as compared with 85 pS in 1 M KCl) with a slight decrease in cation selectivity, indicating that the Φ-clamp is highly conserved and crucial for binary toxin activity. In contrast, the mutants Q367D, N430D, L443E had no or only minor effects on Ib properties, while T360I, T360A and T360W caused a dramatic effect on ion selectivity and single-channel conductance, indicating gross disturbance of the oligomer structure. This suggests that, at least in the iota toxin family, T360 has a structural role in the pore organization. Moreover, introduction of charged residues within the channel (S339E-S341E) or in the vestibule (Q367D, N430D and L443E) had virtually no effect on chloroquine or Ia binding, whereas F454A, T360I, T360A and T360W strongly decreased the chloroquine and Ia affinity to Ib. These results support that distinct residues within the vestibule interact with chloroquine and Ia or are responsible for channel structure, while the channel lining amino acids play a less important role.

  12. Light-activated, in situ forming gel for sustained suprachoroidal delivery of bevacizumab.

    PubMed

    Tyagi, Puneet; Barros, Matthew; Stansbury, Jeffrey W; Kompella, Uday B

    2013-08-05

    A light-activated polycaprolactone dimethacrylate (PCM) and hydroxyethyl methacrylate (HEMA) based gel network was developed to sustain the release of stable, active bevacizumab (an anti-VEGF antibody used to treat choroidal neovascularization) and used to assess sustained ex vivo delivery in rabbit eyes and in vivo delivery in rat eyes following in situ gel formation in the suprachoroidal space. PCM was synthesized from polycaprolactone diol (PCD) and evaluated using NMR spectroscopy. PCM was used to cross-link HEMA in the presence of 365 nm UV light and 2,2-dimethoxy-2-phenylacetophenone (DMPA) as a photoinitiator. Bevacizumab was entrapped in the gel using three different cross-linking durations of 3, 7, and 10 min. In vitro release of bevacizumab in PBS pH 7.4 at 37 °C during a 4 month study was quantified using a VEGF-binding based ELISA. The stability of released bevacizumab was monitored by size exclusion chromatography (SEC) and circular dichroism. Alexa Fluor 488 dye conjugated bevacizumab mixed with polymers was injected suprachoroidally in rabbit eyes to study the effect of different cross-linking durations on the spread of the dye conjugated bevacizumab. In vivo delivery was assessed in Sprague-Dawley (SD) rats by injecting Alexa Fluor 488 dye conjugated bevacizumab mixed with polymers followed by cross-linking for 10 min. Spread in the rabbit eyes and in vivo delivery in rat eyes was monitored noninvasively using a fundus camera and Fluorotron Master. The formation of PCM was confirmed by the disappearance of hydroxyl peak in NMR spectra. A cross-linking duration of 10 min resulted in a burst release of 21% of bevacizumab. Other cross-linking durations had ≥62% burst release. Bevacizumab release from 10 min cross-linked gel was sustained for ∼4 months. Release samples contained ≥96.1% of bevacizumab in the monomeric form as observed in SEC chromatograms. Circular dichroism confirmed that secondary β-sheet structure of bevacizumab was maintained

  13. 76 FR 28239 - Agency Information Collection Activities: Arrival and Departure Record (Forms I-94 and I-94W) and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-16

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  14. Active form of AKT controls cell proliferation and response to apoptosis in hepatocellular carcinoma

    PubMed Central

    KUNTER, IMGE; ERDAL, ESRA; NART, DENIZ; YILMAZ, FUNDA; KARADEMIR, SEDAT; SAGOL, OZGUL; ATABEY, NESE

    2014-01-01

    Hepatocellular carcinoma (HCC) is the third most common cause of cancer-related mortality worldwide. Deregulation of the AKT signaling pathway has been found in HCC. However, the effect of AKT activation on the proliferation and apoptosis in HCC is not clear. Herein, expression of phosphorylated form of AKT (Ser 473) was investigated in HCC tumor (n=73), cirrhosis (n=17), normal liver (n=22) samples and in HCC cell lines (n=8). The results showed that expression of p-AKT was higher in tumor (53%) than in cirrhotic tissues (12%) while it was absent in normal liver (p<0.0001). p-AKT expression was also associated with number of tumor nodules and differentiation status (p<0.05). LY294002 induced cell cycle arrest at G0/G1 in SNU-449 and Mahlavu cells by decreasing expression of CDK2, CDK4, CycD1, CycD3, CycE, CycA and increasing expression of p21 and p27 as well; it also caused a decrease in the E2F1 transcriptional activity through declining phosphorylated Rb. LY294002 did not affect the basal level of apoptosis; however, it amplified cisplatin-induced apoptosis in SNU-449 cells. When the p-AKT level was decreased specifically after transfection with the DN-AKT plasmid, SNU-449 cells became more sensitive to cisplatin-induced apoptosis. HuH-7 cells with no basal p-AKT, were markedly affected by the treatment of doxorubicin. Thus, Akt signaling controls growth and chemical-induced apoptosis in HCC and p-AKT may be a potential target for therapeutic interventions in HCC patients. PMID:24337632

  15. [Sports activity as a form of increasing efficiency in rehabilitation of handicapped people].

    PubMed

    Sobiecka, Joanna

    2002-01-01

    The aim of the research was to find out whether systematic sport training by disabled, fulfills its therapeutic aim in rehabilitation process and gives them back a place in healthy people community. Polish paralympians who took active part in Paralympic Games in years 1992-1998 were questioned (group I). 89% of the respondents of the whole community filled in the questionnaire. The data were compared to the results of the researches among participants of 1972-1988 Paralympic games (group II). The analysis of the influence of systematic training shown that 70% of athletes group I and II, except training, did not have any treatment and sport for 81% from group I and 94% from group II was the only form of rehabilitation and continuation of that process. In spite of professional practising sport, none of paralympians from 1992-1998 noticed worsening of their general health state. Thanks to training, paralympians accepted their disability, and for 62% of respondents in group I and 70% in group II, it was not a barrier to get married. Furthermore, one showed that sport was an agent helping to get better education (23% in years 1992-1998 and 16% in years 1972-1988). 65% respondents (group I) and 60% (group II) assumed that the experience which they got by sport, is helpful in occupational or school duties. The results suggest a need of showing disabled people--if there are no contraindications to physical activity--as early as possible even on hospital rehabilitation state, possibility of continuation of rehabilitation by sport, according to their own interests and physical ability. Medical personnel working in health service should be obliged to qualify patients to training groups, and giving them information on the place where training take part.

  16. 78 FR 2415 - Agency Information Collection Activities: Application for Family Unity Benefits, Form Number I...

    Federal Register 2010, 2011, 2012, 2013, 2014

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  17. 76 FR 69274 - Agency Information Collection Activities: Form I-817, Extension of a Currently Approved...

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    2011-11-08

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  18. 77 FR 21105 - Agency Information Collection Activities: Form G-1145, Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-09

    ... their cell phone that their application or petition has been accepted, they are requested to provide their email address and/or cell phone number on Form G-1145, and attach the form to the application...

  19. 76 FR 51997 - Agency Information Collection Activities: Form I-602; Extension of an Existing Information...

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    2011-08-19

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  20. 76 FR 24908 - Agency Information Collection Activities: Form I-693, Revision of a Currently Approved...

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    ... Information Collection Under Review: Form I- 693, Report of Medical Examination and Vaccination Record, OMB... Vaccination Record. (3) Agency form number, if any, and the applicable component of the Department of...

  1. 76 FR 41282 - Agency Information Collection Activities: Form I-693, Revision of a Currently Approved...

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    2011-07-13

    ... Information Collection Under Review: Form I- 693, Report of Medical Examination and Vaccination Record. The.../Collection: Report of Medical Examination and Vaccination Record. (3) Agency form number, if any, and...

  2. 77 FR 36285 - Agency Information Collection Activities: Form I-693, Revision of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-18

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  3. 75 FR 65500 - Agency Information Collection Activities: Form I-360, Extension of a Currently Approved...

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  4. 76 FR 45844 - Agency Information Collection Activities: Form N-426, Extension of a Currently Approved...

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    2011-08-01

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  5. 76 FR 27078 - Agency Information Collection Activities: Form N-426, Extension of a Currently Approved...

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  6. 77 FR 33758 - Agency Information Collection Activities: Form I-829, Extension of a Currently Approved...

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  8. 75 FR 35003 - Commission Information Collection Activities (FERC Form No. 580); Request; Submitted for OMB...

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    2010-02-01

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  11. 75 FR 11192 - Agency Information Collection Activities: Form I-730, Revision of an Existing Information...

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    2010-03-10

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  13. 75 FR 71451 - Agency Information Collection Activities: Form I-130, Extension of a Currently Approved...

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  14. 75 FR 37820 - Agency Information Collection Activities: Form I-360, Extension of an Existing Information...

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    ... Collection Under Review: Form I- 360, Petition for Amerasian, Widow, or Special Immigrant. OMB Control No... 83-C, Correction Worksheet to amend the instructions to the Form I-360 to add special immigrant... information collection. (2) Title of the Form/Collection: Petition for Amerasian, Widow or Special...

  15. The Main Sequences of Star-forming Galaxies and Active Galactic Nuclei at High Redshift

    NASA Astrophysics Data System (ADS)

    Mancuso, C.; Lapi, A.; Shi, J.; Cai, Z.-Y.; Gonzalez-Nuevo, J.; Béthermin, M.; Danese, L.

    2016-12-01

    We provide a novel, unifying physical interpretation on the origin, average shape, scatter, and cosmic evolution for the main sequences of star-forming galaxies and active galactic nuclei (AGNs) at high redshift z≳ 1. We achieve this goal in a model-independent way by exploiting: (i) the redshift-dependent star formation rate functions based on the latest UV/far-IR data from HST/Herschel, and related statistics of strong gravitationally lensed sources; (ii) deterministic evolutionary tracks for the history of star formation and black hole accretion, gauged on a wealth of multiwavelength observations including the observed Eddington ratio distribution. We further validate these ingredients by showing their consistency with the observed galaxy stellar mass functions and AGN bolometric luminosity functions at different redshifts via the continuity equation approach. Our analysis of the main sequence for high-redshift galaxies and AGNs highlights that the present data are consistently interpreted in terms of an in situ coevolution scenario for star formation and black hole accretion, envisaging these as local, time-coordinated processes.

  16. Investigating microbial colonization in actively forming hydrothermal deposits using thermocouple arrays

    NASA Astrophysics Data System (ADS)

    Tivey, M. K.; Reysenbach, A. L.; Hirsch, M.; Steinberg, J.; Flores, G. E.

    2010-12-01

    Investigations of microbial colonization of very young hydrothermal deposits were carried out in 2009 at hydrothermal vents in the Lau Basin (SW Pacific), and in Guaymas Basin, Gulf of California, with a test deployment at the Rainbow vent field on the Mid-Atlantic Ridge in 2008. Our method entailed razing active chimneys and placing arrays of temperature probes (8 titanium-encased probes with their tips placed within a titanium cage) over the active flow. The chimneys that grew back through each array, encasing the temperature probe tips, were recovered after 2 to 15 days, along with temperature records. Molecular phylogenetic methods are being used to reveal the members of the microbial communities that developed in each chimney of known age and thermal history. A total of 15 array deployments were made at 10 vents in 6 different vent fields. Similar morphology beehives (with porous fine-grained interiors and steep temperature gradients across the outermost more-consolidated “wall”) formed at 2 of the 3 vents in Guaymas Basin (in 2 and 5 days at one vent and 3 and 15 days at a second), and at one vent each in the Kilo Moana (in 3 days), Tahi Moana (in 2.5 days), and Tui Malila (in 3 and 8 days) vent fields in the Lau Basin. In contrast, open conduit, thin walled chimneys grew within arrays at the Mariner vent field, Lau Basin, at 3 different vents (in 3 days at one vent, in 3 and 11 days at a second vent, and in 13 days at a third vent). A lower temperature (<280C) diffuser/spire with a filamentous biofilm formed in 15 days in an array at a hydrocarbon-rich vent in the Guaymas Basin. A similar biofilm formed after 6 days within an array placed earlier at this same vent, with little mineralization. Preliminary diversity data from the 6 and 15 day Guaymas deployments show an increased diversity of bacteria with time with initial colonizers being primarily sulfur-oxidizing Epsilonproteobacteria, with members of the Aquificales and Deltaproteobacteria appearing

  17. How Much Structuring Is Beneficial with Regard to Examination Scores? A Prospective Study of Three Forms of Active Learning

    ERIC Educational Resources Information Center

    Reinhardt, Claus H.; Rosen, Evelyne N.

    2012-01-01

    Many studies have demonstrated a superiority of active learning forms compared with traditional lecture. However, there is still debate as to what degree structuring is necessary with regard to high exam outcomes. Seventy-five students from a premedical school were randomly attributed to an active lecture group, a cooperative group, or a…

  18. In contrast to agonist monoclonal antibodies, both C-terminal truncated form and full length form of Pleiotrophin failed to activate vertebrate ALK (anaplastic lymphoma kinase)?

    PubMed

    Mathivet, Thomas; Mazot, Pierre; Vigny, Marc

    2007-12-01

    Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase essentially and transiently expressed during development in specific regions of the central and peripheral nervous system. ALK expression persists at a lower level in the adult brain. Thus, it might play an important role in both the normal development and function of the nervous system. The nature of the cognate ligand of this receptor in vertebrates is still a matter of debate. Pleiotrophin and midkine have been proposed as ligands of ALK but several independent studies do not confirm this hypothesis. Interestingly, a recent study proposed that a C-terminal truncated form of Pleiotrophin (Pleiotrophin.15) and not the full length form (Pleiotrophin.18) promotes glioblastoma proliferation in an ALK-dependent fashion. These data were obviously a strong basis to conciliate the conflicting results so far reported in the literature. In the present study, we first purified to homogeneity the two forms of Pleiotrophin secreted by HEK 293 cells. In contrast to agonist monoclonal antibodies, both Pleiotrophin.15 and Pleiotrophin.18 failed to activate ALK in neuroblastoma and glioblastoma cells expressing this receptor. Thus, for our point of view, ALK is still an orphan receptor in vertebrates.

  19. Active form Notch4 promotes the proliferation and differentiation of 3T3-L1 preadipocytes

    SciTech Connect

    Lai, Peng-Yeh; Tsai, Chong-Bin; Tseng, Min-Jen

    2013-01-18

    Highlights: ► Notch4IC modulates the ERK pathway and cell cycle to promote 3T3-L1 proliferation. ► Notch4IC facilitates 3T3-L1 differentiation by up-regulating proadipogenic genes. ► Notch4IC promotes proliferation during the early stage of 3T3-L1 adipogenesis. ► Notch4IC enhances differentiation during subsequent stages of 3T3-L1 adipogenesis. -- Abstract: Adipose tissue is composed of adipocytes, which differentiate from precursor cells in a process called adipogenesis. Many signal molecules are involved in the transcriptional control of adipogenesis, including the Notch pathway. Previous adipogenic studies of Notch have focused on Notch1 and HES1; however, the role of other Notch receptors in adipogenesis remains unclear. Q-RT-PCR analyses showed that the augmentation of Notch4 expression during the differentiation of 3T3-L1 preadipocytes was comparable to that of Notch1. To elucidate the role of Notch4 in adipogenesis, the human active form Notch4 (N4IC) was transiently transfected into 3T3-L1 cells. The expression of HES1, Hey1, C/EBPδ and PPARγ was up-regulated, and the expression of Pref-1, an adipogenic inhibitor, was down-regulated. To further characterize the effect of N4IC in adipogenesis, stable cells expressing human N4IC were established. The expression of N4IC promoted proliferation and enhanced differentiation of 3T3-L1 cells compared with those of control cells. These data suggest that N4IC promoted proliferation through modulating the ERK pathway and the cell cycle during the early stage of 3T3-L1 adipogenesis and facilitated differentiation through up-regulating adipogenic genes such as C/EBPα, PPARγ, aP2, LPL and HSL during the middle and late stages of 3T3-L1 adipogenesis.

  20. DNA Recombinase Proteins, their Function and Structure in the Active Form, a Computational Study

    NASA Technical Reports Server (NTRS)

    Carra, Claudio; Cucinotta, Francis A.

    2007-01-01

    Homologous recombination is a crucial sequence of reactions in all cells for the repair of double strand DNA (dsDNA) breaks. While it was traditionally considered as a means for generating genetic diversity, it is now known to be essential for restart of collapsed replication forks that have met a lesion on the DNA template (Cox et al., 2000). The central stage of this process requires the presence of the DNA recombinase protein, RecA in bacteria, RadA in archaea, or Rad51 in eukaryotes, which leads to an ATP-mediated DNA strand-exchange process. Despite many years of intense study, some aspects of the biochemical mechanism, and structure of the active form of recombinase proteins are not well understood. Our theoretical study is an attempt to shed light on the main structural and mechanistic issues encountered on the RecA of the e-coli, the RecA of the extremely radio resistant Deinococcus Radiodurans (promoting an inverse DNA strand-exchange repair), and the homolog human Rad51. The conformational changes are analyzed for the naked enzymes, and when they are linked to ATP and ADP. The average structures are determined over 2ns time scale of Langevian dynamics using a collision frequency of 1.0 ps(sup -1). The systems are inserted in an octahedron periodic box with a 10 Angstrom buffer of water molecules explicitly described by the TIP3P model. The corresponding binding free energies are calculated in an implicit solvent using the Poisson-Boltzmann solvent accessible surface area, MM-PBSA model. The role of the ATP is not only in stabilizing the interaction RecA-DNA, but its hydrolysis is required to allow the DNA strand-exchange to proceed. Furthermore, we extended our study, using the hybrid QM/MM method, on the mechanism of this chemical process. All the calculations were performed using the commercial code Amber 9.

  1. Broca's region and Visual Word Form Area activation differ during a predictive Stroop task.

    PubMed

    Wallentin, Mikkel; Gravholt, Claus Højbjerg; Skakkebæk, Anne

    2015-12-01

    Competing theories attempt to explain the function of Broca's area in single word processing. Studies have found the region to be more active during processing of pseudo words than real words and during infrequent words relative to frequent words and during Stroop (incongruent) color words compared to Non-Stroop (congruent) words. Two related theories explain these findings as reflecting either "cognitive control" processing in the face of conflicting input or a linguistic prediction error signal, based on a predictive coding approach. The latter implies that processing cost refers to violations of expectations based on the statistical distributions of input. In this fMRI experiment we attempted to disentangle single word processing cost originating from cognitive conflict and that stemming from predictive expectation violation. Participants (N = 49) responded to whether the words "GREEN" or "RED" were displayed in green or red (incongruent vs congruent colors). One of the colors, however, was presented three times as often as the other, making it possible to study both congruency and frequency effects independently. Auditory stimuli saying "GREEN" or "RED" had the same distribution, making it possible to study frequency effects across modalities. We found significant behavioral effects of both incongruency and frequency. A significant effect (p < .05 FWE) of incongruency was found in Broca's region, but no effect of frequency was observed and no interaction. Conjoined effects of incongruency and frequency were found in parietal regions as well as in the Visual Word Form Area (VWFA). No interaction between perceptual modality and frequency was found in VWFA suggesting that the region is not strictly visual. These findings speak against a strong version of the prediction error processing hypothesis in Broca's region. They support the idea that prediction error processes in the intermediate timeframe are allocated to more posterior parts of the brain.

  2. Comparison between two forms of granular activated carbon for the removal of pharmaceuticals from different waters.

    PubMed

    Lima, Lisandra; Baêta, Bruno E L; Lima, Diego R S; Afonso, Robson J C F; de Aquino, Sérgio F; Libânio, Marcelo

    2016-01-01

    The aim of this study was to evaluate the performance of two forms of basic granular activated carbon (GAC), mineral (pH = 10.5) and vegetal (pH = 9), for the removal of three pharmaceuticals, as sulphamethoxazole (SMX), diclofenac (DCF) and 17β-estradiol (E2), from two different matrices: fortified distilled (2.4-3.0 mg L(-1) and pH from 5.5 to 6.5) and natural (∼1.0 mg L(-1) and pH from 7.1 to 7.2) water in a bench scale. The Rapid Small-Scale Column Test used to assess the ability of mineral and vegetal GAC on removal of such pharmaceuticals led to removal capacities varying from 14.9 to 23.5 mg g(-1) for E2, from 23.7 to 24.2 mg g(-1) for DCF and from 20.5 to 20.6 mg g(-1) for SMX. Removal efficiencies of 71%, 88% and 74% for DCF, SMX and E2, respectively, were obtained at breakthrough point when using mineral GAC, whereas for the vegetal GAC the figures were 76%, 77% and 65%, respectively. The carbon usage rate at the breakthrough point varied from 11.9 to 14.5 L g(-1) for mineral GAC and from 8.8 to 14.8 L g(-1) for vegetal GAC. Mineral CAG also exhibited the best performance when treating fortified natural water, since nearly complete removal was observed for all contaminants in the column operated for 22 h at a carbon usage rate of 2.9 L g(-1).

  3. Expression of protease-activated receptor 1 and 2 and anti-tubulogenic activity of protease-activated receptor 1 in human endothelial colony-forming cells.

    PubMed

    Fortunato, Tiago M; Vara, Dina S; Wheeler-Jones, Caroline P; Pula, Giordano

    2014-01-01

    Endothelial colony-forming cells (ECFCs) are obtained from the culture of human peripheral blood mononuclear cell (hPBMNC) fractions and are characterised by high proliferative and pro-vasculogenic potential, which makes them of great interest for cell therapy. Here, we describe the detection of protease-activated receptor (PAR) 1 and 2 amongst the surface proteins expressed in ECFCs. Both receptors are functionally coupled to extracellular signal-regulated kinase (ERK) 1 and 2, which become activated and phosphorylated in response to selective PAR1- or PAR2-activating peptides. Specific stimulation of PAR1, but not PAR2, significantly inhibits capillary-like tube formation by ECFCs in vitro, suggesting that tubulogenesis is negatively regulated by proteases able to stimulate PAR1 (e.g. thrombin). The activation of ERKs is not involved in the regulation of tubulogenesis in vitro, as suggested by use of the MEK inhibitor PD98059 and by the fact that PAR2 stimulation activates ERKs without affecting capillary tube formation. Both qPCR and immunoblotting showed a significant downregulation of vascular endothelial growth factor 2 (VEGFR2) in response to PAR1 stimulation. Moreover, the addition of VEGF (50-100 ng/ml) but not basic Fibroblast Growth Factor (FGF) (25-100 ng/ml) rescued tube formation by ECFCs treated with PAR1-activating peptide. Therefore, we propose that reduction of VEGF responsiveness resulting from down-regulation of VEGFR2 is underlying the anti-tubulogenic effect of PAR1 activation. Although the role of PAR2 remains elusive, this study sheds new light on the regulation of the vasculogenic activity of ECFCs and suggests a potential link between adult vasculogenesis and the coagulation cascade.

  4. 75 FR 30098 - Reports, Forms and RecordKeeping Requirements; Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

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  1. 76 FR 60852 - Agency Information Collection Activities; Form I-130; Extension of an Existing Information...

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  4. Effect of parental age and developmental rate on the production of active form of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae).

    PubMed

    Sano-Fujii, I

    1979-05-01

    Three similar experiments were conducted to see the effect of parental age and parental developmental speed on the production of "active-form" (i.e., capable of flight) offspring in Callosobruchus maculatus (F.). In experiment I, using the adults which emerged on three different days (as fast, intermediate, and slow in development), eggs were collected on three different days (at young, middle, and old age) during the adult life span. The results showed that the older parents produced more active-form offspring irrespective of the parents' own rate of development. In experiment II, the parental rate of development was investigated in more detail by using the adults that emerged over the entire span of their emergence period. In experiment III, the effect of parental age was investigated in more detail by collecting the eggs daily during the female adult's entire life span. Both experiments confirmed the trend observed in experiment I. From these results it can be concluded that the older the parents, the higher the percentage of the active form in their offspring. However, there was no significant systematic trend due to the length of the developmental period of the parents as seen in the parental age. This trend, that more active forms emerge from older parents, seems to be related to the decreased viability of older parents. It is thought that the less viable eggs produced by older parents are more susceptible to stimuli which induce the active form.

  5. Application of a heterogeneous immunoassay for the quality control testing of release-active forms of diclofenac.

    PubMed

    Pschenitza, Michael; Gavrilova, Elena S; Tarasov, Sergey А; Knopp, Dietmar; Niessner, Reinhard; Epstein, Oleg I

    2014-07-01

    We report on a specially designed diclofenac-ELISA for the determination of diclofenac in the presence of release-active forms of diclofenac in lactose dissolved in water solutions according to a predefined schedule in single-blind experiments. In accordance with the objective of this project, a number of experiments were conducted to determine the optimal ELISA conditions for detecting potential modulatory effects of release-active forms of diclofenac depending on their ability to affect the binding of diclofenac to anti-diclofenac antibodies. As a feature, the diclofenac antibodies were previously incubated with manufactured pharmaceutical samples containing release-active forms of diclofenac or placebo. For comparison of the sample types, measured in ELISA optical densities were chosen. For statistic analysis, Student's two-sample t-test and single-factor ANOVA were applied. The extremely low concentrations of diclofenac of 0.01, 0.05 and 0.1 ng mL(-1) seem most appropriate for routine assay performance. The source of diclofenac used for standard solution preparation is not important but it could be important as the source of diclofenac for release active form of diclofenac preparation. As an outcome, the ELISA appeared to be suitable for the detection of the modifying effects of release-active forms of diclofenac toward the pharmaceutical substance in vitro.

  6. DEVELOPMENT QUALIFICATION AND DISPOSAL OF AN ALTERNATIVE IMMOBILIZED LOW-ACTIVITY WASTE FORM AT THE HANFORD SITE

    SciTech Connect

    SAMS TL; EDGE JA; SWANBERG DJ; ROBBINS RA

    2011-01-13

    Demonstrating that a waste form produced by a given immobilization process is chemically and physically durable as well as compliant with disposal facility acceptance criteria is critical to the success of a waste treatment program, and must be pursued in conjunction with the maturation of the waste processing technology. Testing of waste forms produced using differing scales of processing units and classes of feeds (simulants versus actual waste) is the crux of the waste form qualification process. Testing is typically focused on leachability of constituents of concern (COCs), as well as chemical and physical durability of the waste form. A principal challenge regarding testing immobilized low-activity waste (ILAW) forms is the absence of a standard test suite or set of mandatory parameters against which waste forms may be tested, compared, and qualified for acceptance in existing and proposed nuclear waste disposal sites at Hanford and across the Department of Energy (DOE) complex. A coherent and widely applicable compliance strategy to support characterization and disposal of new waste forms is essential to enhance and accelerate the remediation of DOE tank waste. This paper provides a background summary of important entities, regulations, and considerations for nuclear waste form qualification and disposal. Against this backdrop, this paper describes a strategy for meeting and demonstrating compliance with disposal requirements emphasizing the River Protection Project (RPP) Integrated Disposal Facility (IDF) at the Hanford Site and the fluidized bed steam reforming (FBSR) mineralized low-activity waste (LAW) product stream.

  7. A study of renaturation of reduced hen egg white lysozyme. Enzymically active intermediates formed during oxidation of the reduced protein.

    PubMed

    Acharya, A S; Taniuchi, H

    1976-11-25

    The material obtained from reduced hen egg white lysozyme after complete air oxidation at pH 8.0 and 37 degrees has yielded, by gel filtration on a Bio-Gel P-30 column, enzymically active species and an enzymically inactive form which eluted sooner than the active species but later than expected for a dimer of lysozyme. Reduced lysozyme also elutes at the same position as this inactive material. Examination of the fragments produced on CNBr cleavage of the inactive form indicates that at least 24% of the population contains incorrect disulfide bonds involving half-cystine residues 6, 30, 115, and 127. Tryptophan fluorescence and the intrinsic viscosity of the inactive form show an enlarged molecular domain with a disordered conformation. The yield of the inactive form increases as the oxidation of reduced lysozyme is accelerated using cupric ion. In the presence of 4 X 10(-5) M cupric ion, reduced lysozyme forms almost quantitatively the inactive form, which is almost completely converted to the native form by sulfhydryl-disulfide interchange catalyzed by thiol groups of either reduced lysozyme or beta-mercaptoethanol. The material trapped by alkylation of the free sulfhydryl groups with [1-14C]iodoacetic acid during the early stage of air oxidation of reduced lysozyme was fractionated by gel filtration to permit separation of the active species from the inactive form. Ion exchange chromatography of the active species yielded completely renatured lysozyme and three major enzymically active radioactive derivatives. Two of these derivatives contained approximately 2 mol of S-carboxymethylcysteine. Isolation and characterization of radioactive tryptic peptides from each of the three active forms, permitted the identification of Cys 6 and Cys 127, Cys 76 and 94, and Cys 80 as the sulfhydryl groups alkylated in these three incompletely oxidized, partially active forms. Thus, it appears that the interatomic interactions maintaining the compact three-dimensional structure

  8. Observational analysis of the physical conditions in galactic and extragalactic active star forming regions

    NASA Astrophysics Data System (ADS)

    Kristensen, L. E.

    2007-10-01

    In my thesis observations of near-infrared rovibrational H_2 emission in active star-forming regions are presented and analysed. The main subject of this work concerns new observations of the Orion Molecular Cloud (OMC1) and in particular the BN-KL region. Data consist of images of individual H_2 lines with high spatial resolution obtained both at the Canada-France-Hawaii Telescope and at the ESO Very Large Telescope (VLT). With the high spatial resolution of the VLT it is possible to analyse in detail (down to 60 AU ~ 0.13") individual objects in the region. I have also analysed H_2 and [FeII] emission from outflows in two dark clouds (Bok globules BHR71 and BHR137) and a high excitation blob in the Magellanic Clouds (N159-5). In the latter, data consist of long-slit spectra obtained at the ESO-VLT. In order to facilitate this work I ran a large grid of ~25000 shock models, producing almost 400 Gb of results. These models are state-of-the-art and there is a large number of free parameters which can be adjusted. A big part of my project has been to analyse the results from this grid and make it publically available. Furthermore, as it turned out, not all results are equally reliable and I have had to develop methods for checking the consistency of the wealth of results obtained. But with the model results and a sound knowledge of shock physics it is now relatively straightforward to interpret the H_2 and [FeII] data. The models allow me to predict the large-scale physical conditions in OMC1 such as density, shock velocities, magnetic field strengths, etc. Overall the preshock density is of the order of ~10^5-10^7 cm(-3) and shock velocities are in the interval 10-40 km/s. Another very interesting result is a new method developed for analysing bow shocks observed at high spatial resolution. For one isolated bow shock in OMC1 I predict a shock velocity of 50 km/s and a preshock density of the order of 5x10^5 cm(-3). The 3D velocity has recently been measured to 55 km

  9. 40 CFR 761.205 - Notification of PCB waste activity (EPA Form 7710-53).

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... handling activity and now wishes to engage in another PCB waste activity (e.g., previously only... 40 Protection of Environment 31 2014-07-01 2014-07-01 false Notification of PCB waste activity..., DISTRIBUTION IN COMMERCE, AND USE PROHIBITIONS PCB Waste Disposal Records and Reports § 761.205 Notification...

  10. 40 CFR 761.205 - Notification of PCB waste activity (EPA Form 7710-53).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... handling activity and now wishes to engage in another PCB waste activity (e.g., previously only... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Notification of PCB waste activity..., DISTRIBUTION IN COMMERCE, AND USE PROHIBITIONS PCB Waste Disposal Records and Reports § 761.205 Notification...

  11. Advantages of Gardening as a Form of Physical Activity in an After-School Program

    ERIC Educational Resources Information Center

    Phelps, Joshua; Hermann, Janice R.; Parker, Stephany P.; Denney, Barbara

    2010-01-01

    Children who normally abstain from physical activity may view gardening as a viable non-competitive alternative. The study reported here evaluated the effect of an Oklahoma Cooperative Extension Service after-school gardening program on self-reported physical activity level of children in 3rd through 5th grade using the ACTIVITY self-report…

  12. Physicochemical properties and anti-Propionibacterium acnes activity of film-forming solutions containing alpha-mangostin-rich extract.

    PubMed

    Asasutjarit, Rathapon; Larpmahawong, Papassara; Fuongfuchat, Asira; Sareedenchai, Vipaporn; Veeranondha, Sukitaya

    2014-04-01

    The objective of this study was to study the effect of formulation compositions on physicochemical properties and anti-Propionibacterium acnes activity of film-forming solutions containing alpha-mangostin-rich extract (AM). Film-forming solution bases and film-forming solutions containing AM were prepared by using Eudragit RL PO or Klucel LF or combinations of them as film-forming polymers. Rheological properties, pH values of the solutions, and mechanical properties of the dry films were investigated. An optimized formulation was selected and evaluated for the film surface, in vitro AM release, an anti-P. acnes activity, and potential for being a skin irritant. It was found that mechanical properties of the dry films were affected by total polymer contents, ratios of Klucel LF/Eudragit RL PO, AM, and contents of triethyl citrate. The film-forming solutions containing AM had pH values around 7.0. Their flow curves exhibited Newtonian flow behaviors. The optimized formulation provided films possessing smooth and nonporous surfaces. These films showed greater anti-P. acnes activity than their base films without toxicity to skin fibroblasts. Furthermore, AM released from the film matrix obeyed Higuchi's equation. In conclusion, the film-forming solutions containing AM had potential for treatment of acne vulgaris caused by P. acnes. However, further in vivo study is necessary to determine their efficacy and safety for using in patients suffering from acne vulgaris.

  13. Film-forming formulations containing porous silica for the sustained delivery of actives to the skin.

    PubMed

    Heck, Rouven; Hermann, Sabrina; Lunter, Dominique J; Daniels, Rolf

    2016-11-01

    The purpose of this study was to develop film-forming formulations facilitating long-term treatment of chronic pruritus with capsaicinoids. To this end, an oily solution of nonivamide was loaded into porous silica particles which were then suspended in the dispersion of a sustained release polymer. Such formulations form a film when applied to the skin and encapsulate the drug loaded silica particles in a dry polymeric matrix. Dermal delivery and permeation of the antipruritic drug nonivamide (NVA) are controlled by the matrix. The film-forming formulations were examined regarding homogeneity, storage stability, substantivity and ex vivo skin permeation. Confocal Raman spectral imaging proved the stability of silica-based film-forming formulations over a period of 6 months. Substantivity was found to be enhanced substantially compared to a conventional semisolid formulation. Permeation rates of nonivamide from film-forming formulations through the skin are much lower compared to those achieved with a conventional immediate release formulation with the same drug amount. Due to the drug reservoir in the polymer matrix, a sustained permeation is enabled. Film-forming formulations may therefore improve the treatment of chronic pruritus with capsaicinoids by enhancing patient compliance through a sustained release regime.

  14. Structural data suggest that the active and inactive forms of the RecA filament are not simply interconvertible.

    PubMed

    Yu, X; Egelman, E H

    1992-09-05

    We have used electron microscopy to examine the two major conformational states of the helical filament formed by the RecA protein of Escherichia coli. The compressed filament, formed in the absence of a nucleotide cofactor either as a self-polymer or on a single-stranded DNA molecule, is characterized in solution by about 6.1 subunits per turn of a 76 A pitch helix, and appears to be inactive with respect to all RecA activity. The active state of the filament, formed with ATP or an ATP analog on either a single or double-stranded DNA substrate, has about 6.2 subunits per turn of a 94 A pitch helix. Measurements of the contour length of RecA-covered single-stranded DNA circles in ice, formed in the absence of nucleotide cofactor, indicate that each RecA subunit binds five bases, in contrast to the three bases or base-pairs per subunit in the active state. The different stoichiometries of DNA binding suggests that the two polymeric forms are not interconvertible, as has been suggested on biochemical grounds. A three-dimensional reconstruction of the inactive state shows the same general features as the 83 A pitch filament present in the RecA crystal. This structural similarity and the fact that the crystal does not contain ATP or DNA suggests that the crystal structure is more similar to the compressed filament than the active, extended filament.

  15. Pathogenic forms of tau inhibit kinesin-dependent axonal transport through a mechanism involving activation of axonal phosphotransferases.

    PubMed

    Kanaan, Nicholas M; Morfini, Gerardo A; LaPointe, Nichole E; Pigino, Gustavo F; Patterson, Kristina R; Song, Yuyu; Andreadis, Athena; Fu, Yifan; Brady, Scott T; Binder, Lester I

    2011-07-06

    Aggregated filamentous forms of hyperphosphorylated tau (a microtubule-associated protein) represent pathological hallmarks of Alzheimer's disease (AD) and other tauopathies. While axonal transport dysfunction is thought to represent a primary pathogenic factor in AD and other neurodegenerative diseases, the direct molecular link between pathogenic forms of tau and deficits in axonal transport remain unclear. Recently, we demonstrated that filamentous, but not soluble, forms of wild-type tau inhibit anterograde, kinesin-based fast axonal transport (FAT) by activating axonal protein phosphatase 1 (PP1) and glycogen synthase kinase 3 (GSK3), independent of microtubule binding. Here, we demonstrate that amino acids 2-18 of tau, comprising a phosphatase-activating domain (PAD), are necessary and sufficient for activation of this pathway in axoplasms isolated from squid giant axons. Various pathogenic forms of tau displaying increased exposure of PAD inhibited anterograde FAT in squid axoplasm. Importantly, immunohistochemical studies using a novel PAD-specific monoclonal antibody in human postmortem tissue indicated that increased PAD exposure represents an early pathogenic event in AD that closely associates in time with AT8 immunoreactivity, an early marker of pathological tau. We propose a model of pathogenesis in which disease-associated changes in tau conformation lead to increased exposure of PAD, activation of PP1-GSK3, and inhibition of FAT. Results from these studies reveal a novel role for tau in modulating axonal phosphotransferases and provide a molecular basis for a toxic gain-of-function associated with pathogenic forms of tau.

  16. Life form and water source interact to determine active time and environment in cryptogams: an example from the maritime Antarctic.

    PubMed

    Schlensog, Mark; Green, T G Allan; Schroeter, Burkhard

    2013-09-01

    Antarctica, with its almost pristine conditions and relatively simple vegetation, offers excellent opportunities to investigate the influence of environmental factors on species performance, such information being crucial if the effects of possible climate change are to be understood. Antarctic vegetation is mainly cryptogamic. Cryptogams are poikilohydric and are only metabolically and photosynthetically active when hydrated. Activity patterns of the main life forms present, bryophytes (10 species, ecto- and endohydric), lichens (5 species) and phanerogams (2 species), were monitored for 21 days using chlorophyll a fluorescence as an indicator of metabolic activity and, therefore, of water regime at a mesic (hydration by meltwater) and a xeric (hydration by precipitation) site on Léonie Island/West Antarctic Peninsula (67°36'S). Length of activity depended mainly on site and form of hydration. Plants at the mesic site that were hydrated by meltwater were active for long periods, up to 100 % of the measurement period, whilst activity was much shorter at the xeric site where hydration was entirely by precipitation. There were also differences due to life form, with phanerogams and mesic bryophytes being most active and lichens generally much less so. The length of the active period for lichens was longer than in continental Antarctica but shorter than in the more northern Antarctic Peninsula. Light intensity when hydrated was positively related to the length of the active period. High activity species were strongly coupled to the incident light whilst low activity species were active under lower light levels and essentially uncoupled from incident light. Temperatures were little different between sites and also almost identical to temperatures, when active, for lichens in continental and peninsular Antarctica. Gradients in vegetation cover and growth rates across Antarctica are, therefore, not likely to be due to differences in temperature but more likely to

  17. An activated form of ADAM10 is tumor selective and regulates cancer stem-like cells and tumor growth

    PubMed Central

    Saha, Nayanendu; Eissman, Moritz F.; Xu, Kai; Llerena, Carmen; Kusebauch, Ulrike; Ding, Bi-Sen; Cao, Zhongwei; Rafii, Shahin; Ernst, Matthias; Scott, Andrew M.; Nikolov, Dimitar B.; Lackmann, Martin

    2016-01-01

    The transmembrane metalloprotease ADAM10 sheds a range of cell surface proteins, including ligands and receptors of the Notch, Eph, and erbB families, thereby activating signaling pathways critical for tumor initiation and maintenance. ADAM10 is thus a promising therapeutic target. Although widely expressed, its activity is normally tightly regulated. We now report prevalence of an active form of ADAM10 in tumors compared with normal tissues, in mouse models and humans, identified by our conformation-specific antibody mAb 8C7. Structure/function experiments indicate mAb 8C7 binds an active conformation dependent on disulfide isomerization and oxidative conditions, common in tumors. Moreover, this active ADAM10 form marks cancer stem-like cells with active Notch signaling, known to mediate chemoresistance. Importantly, specific targeting of active ADAM10 with 8C7 inhibits Notch activity and tumor growth in mouse models, particularly regrowth after chemotherapy. Our results indicate targeted inhibition of active ADAM10 as a potential therapy for ADAM10-dependent tumor development and drug resistance. PMID:27503072

  18. A facile reflux procedure to increase active surface sites form highly active and durable supported palladium@platinum bimetallic nanodendrites

    NASA Astrophysics Data System (ADS)

    Wang, Qin; Li, Yingjun; Liu, Baocang; Xu, Guangran; Zhang, Geng; Zhao, Qi; Zhang, Jun

    2015-11-01

    A series of well-dispersed bimetallic Pd@Pt nanodendrites uniformly supported on XC-72 carbon black are fabricated by using different capping agents. These capping agents are essential for the branched morphology control. However, the surfactant adsorbed on the nanodendrites surface blocks the access of reactant molecules to the active surface sites, and the catalytic activities of these bimetallic nanodendrites are significantly restricted. Herein, a facile reflux procedure to effectively remove the capping agent molecules without significantly affecting their sizes is reported for activating supported nanocatalysts. More significantly, the structure and morphology of the nanodendrites can also be retained, enhancing the numbers of active surface sites, catalytic activity and stability toward methanol and ethanol electro-oxidation reactions. The as-obtained hot water reflux-treated Pd@Pt/C catalyst manifests superior catalytic activity and stability both in terms of surface and mass specific activities, as compared to the untreated catalysts and the commercial Pt/C and Pd/C catalysts. We anticipate that this effective and facile removal method has more general applicability to highly active nanocatalysts prepared with various surfactants, and should lead to improvements in environmental protection and energy production.

  19. Keep Fit. Suggested Fitness Activities for Forms 1-4 Girls.

    ERIC Educational Resources Information Center

    Dinataci, S.

    This booklet is designed for teachers who conduct physical education programs and who coach sports in schools where modern training equipment is not available. Most of the equipment described or shown in this booklet can be improvised. The activities are designed for girls in grades seven through ten. The following activities are covered in the…

  20. 76 FR 52014 - Agency Information Collection Activities: Applicant Information Form (1-783)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-19

    ... of Investigation (FBI), Criminal Justice Information Services (CJIS) Division will be submitting the... Form); CJIS Division, FBI, DOJ. (4) Affected public who will be asked or required to respond, as well... for an average respondent to respond: Annually, the FBI receives 225,000 identification...

  1. 77 FR 3484 - Agency Information Collection Activities: Form I-914, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-24

    ...] [FR Doc No: 2012-1277] DEPARTMENT OF HOMELAND SECURITY U.S. Citizenship and Immigration Services.... 1615-0099. The Department of Homeland Security, U.S. Citizenship and Immigration Services (USCIS) will... Homeland Security sponsoring the collection: Form I-914. U.S. Citizenship and Immigration Services....

  2. 77 FR 3485 - Agency Information Collection Activities: Form I-129F, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-24

    ...] [FR Doc No: 2012-1354] DEPARTMENT OF HOMELAND SECURITY U.S. Citizenship and Immigration Services..., Petition for Alien Fiance(e). The Department of Homeland Security, U.S. Citizenship and Immigration... collection: Form I-129F. U.S. Citizenship and Immigration Services. (4) Affected public who will be asked...

  3. 77 FR 3486 - Agency Information Collection Activities: Form I-539, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-24

    ... No: 2012-1349] DEPARTMENT OF HOMELAND SECURITY U.S. Citizenship and Immigration Services Agency... Extend/Change Nonimmigrant Status. The Department of Homeland Security, U.S. Citizenship and Immigration... Security sponsoring the collection: Form I-539. U.S. Citizenship and Immigration Services. (4)...

  4. 75 FR 16895 - Reports, Forms and Record Keeping Requirements; Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-02

    ... National Highway Traffic Safety Administration Reports, Forms and Record Keeping Requirements; Agency... Street, NW., Washington, DC 20503, Attention: NHTSA Desk Officer. FOR FURTHER INFORMATION CONTACT: Ms... maintenance costs of keeping the data up to date, etc. as tests come in throughout the year. In the...

  5. 76 FR 35266 - Reports, Forms and Record Keeping Requirements; Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-16

    ... National Highway Traffic Safety Administration Reports, Forms and Record Keeping Requirements; Agency... Management and Budget, 725 17th Street, NW., Washington, DC 20503, Attention NHTSA Desk Officer. Comments are... collection of information on respondents, including the use of automated collection techniques or other...

  6. 78 FR 73237 - Reports, Forms and Record Keeping Requirements Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-05

    ... National Highway Traffic Safety Administration Reports, Forms and Record Keeping Requirements Agency... Regulatory Affairs, Office of Management and Budget, 725-17th Street NW., Washington, DC 20503, Attention... of 49 CFR Part 512. Requesters are not required to keep copies of any records or reports ]...

  7. 78 FR 67212 - Reports, Forms and Record Keeping Requirements; Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-08

    ... National Highway Traffic Safety Administration Reports, Forms and Record Keeping Requirements; Agency... has submitted required information shall keep its entry current, accurate and complete by submitting... Regulatory Affairs, Office of Management and Budget, 725 17th Street NW., Washington, DC 20503,...

  8. 77 FR 56232 - Agency Information Collection Activities; Submission for OMB Review; Comment Request; Forms for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-12

    ... Department of Labor (DOL) is submitting the Employment ] and Training Administration (ETA) sponsored... this request to the Office of Information and Regulatory Affairs, Attn: OMB Desk Officer for DOL-ETA... contractors complete forms ETA-790 (the Agricultural and Food Processing Clearance Order) and ETA-795...

  9. 75 FR 65447 - Agency Information Collection Activities: Proposed Collection; Comment Request Form FNS-648, WIC...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-25

    ... collection, the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) Local Agency..., Director, Supplemental Food Programs Division, Food and Nutrition Service, U.S. Department of Agriculture... Collection Form. Abstract: FNS administers the Special Supplemental Nutrition Program for Women, Infants,...

  10. 76 FR 37059 - Agency Information Collection Activities: Proposed Collection; Comment Request-Form FNS-339...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-24

    ... Request--Form FNS-339, Federal-State Supplemental Nutrition Program(s) Agreement AGENCY: Food and... FNS-339, the Federal and State Agreement for the Special Supplemental Nutrition Program for Women...-2746. SUPPLEMENTARY INFORMATION: Title: Federal-State Supplemental Nutrition Program(s) Agreement....

  11. 76 FR 40386 - Agency Information Collection Activities: Form I-905, Extension of an Existing Information...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-08

    ..., Extension of an Existing Information Collection; Comment Request ACTION: 30-Day Notice of Information Collection Under Review: Form I- 905, Application for Authorization to Issue Certification for Health Care... FR 20362, allowing for a 60-day public comment period. USCIS received one comment after...

  12. 75 FR 74144 - Reports, Forms and Recordkeeping Requirements; Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-30

    .... SUPPLEMENTARY INFORMATION: Maritime Administration (MARAD) Title of Collection: War Risk Insurance, Applications... in MARAD's war risk insurance program. Form Numbers: MA-355; MA-528; MA-742; MA-828, and MA-942... the U.S. Department of Transportation may provide war risk insurance adequate for the needs of...

  13. 77 FR 65705 - Agency Information Collection Activities: Monthly Report on Naturalization Papers, Form Number N...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-30

    ... Report on Naturalization Papers. (3) Agency form number, if any, and the applicable component of the DHS... Collection ACTION: 30-Day Notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and... Officer and to DHS. Comments should be submitted to the OMB USCIS Desk Officer via facsimile at...

  14. 77 FR 65708 - Agency Information Collection Activities: Petition To Remove the Conditions on Residence, Form...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-30

    ... Conditions on Residence. (3) Agency form number, if any, and the applicable component of the DHS sponsoring...: 60-Day Notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration... directed to: DHS, USCIS, Office of Policy and Strategy, Chief, Regulatory Coordination Division,...

  15. 77 FR 67018 - Agency Information Collection Activities: Notice of Naturalization Oath Ceremony, Form Number N...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-11-08

    ... Naturalization Oath Ceremony. (3) Agency form number, if any, and the applicable component of the DHS sponsoring... ACTION: 60-Day Notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and... estimated public burden and associated response time should be directed to: DHS, USCIS, Office of Policy...

  16. 78 FR 5194 - Agency Information Collection Activities: Petition To Remove the Conditions on Residence, Form I...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-24

    ... Conditions on Residence. (3) Agency form number, if any, and the applicable component of the DHS sponsoring...-Day notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration... at 77 FR 65708, allowing for a 60-day public comment period. USCIS did receive 3 comments...

  17. Arousal Model Components in Television Programming: Form Activity and Violent Content.

    ERIC Educational Resources Information Center

    Watt, James H., Jr.; Krull, Robert

    In research reported in this paper, an attempt was made to isolate arousal components due to the "form" of a television program from arousal components due to the "content" of the program. The following hypotheses were formulated: (1) emotional arousal will take place in programing segments depicting violent acts, (2) arousal due to the cognitive…

  18. 78 FR 26766 - Commission Information Collection Activities (FERC Form 580); Comment Request; Revision

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-08

    ... on Fuel and Energy Purchase Practices). DATES: Comments on the collections of information are due.... SUPPLEMENTARY INFORMATION: Title: Interrogatory on Fuel and Energy Purchase Practices (FERC Form No. 580). OMB... Commission collected ``Interrogatory on Fuel and Energy Purchase Practices'' data every other year. \\2\\...

  19. 75 FR 58443 - Agency Information Collection Activities; Submission for OMB Review; Comment Request; Form 5500...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-24

    ... Employee Benefits Security Administration (EBSA) titled, ``Annual Information Return/Report of Employee..._PRA_PUBLIC@dol.gov . SUPPLEMENTARY INFORMATION: The ``Annual Return/Report of Employee Benefit Plan...; Form 5500, Annual Return/Report of Employee Benefit Plan ACTION: Notice. SUMMARY: The Department...

  20. 76 FR 11806 - Agency Information Collection Activities: Form G-1145, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-03

    ... be submitted to: USCIS, Chief, Regulatory Products Division, Office of the Executive Secretariat... notified via e-mail and/or text message on their cell phone that their application or petition has been accepted, they are requested to provide their e-mail address and/or cell phone number on Form G-1145,...

  1. 76 FR 10385 - Agency Information Collection Activities: Various Contract Related Forms That Will be Included in...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-24

    ... Management and Budget (OMB) for review and clearance in accordance with the Paperwork Reduction Act of 1995..., DHS FORM 0700-04 AGENCY: Office of Chief Procurement Officer, Acquisition Policy and Legislation...: The Department of Homeland Security, Office of Chief Procurement Officer, Acquisition Policy...

  2. 77 FR 75112 - Agency Information Collection Activities: Existing Collection; Comment Request-Forms FNS-806-A...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-19

    ...), and FNS-806-B, Claim for Reimbursement (Special Milk Program for Children) AGENCY: Food and Nutrition... Special Milk Program. DATES: Written comments must be received on or before February 19, 2013. ADDRESSES... Lunch Program, Part 220 School Breakfast Program, and Part 215 Special Milk Program. Form Number:...

  3. 77 FR 23734 - Agency Information Collection Activities: Form I-361, Extension of a Currently Approved...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-20

    ... Legal Custody for Public Law 97-359 Amerasian. The Department of Homeland Security, U.S. Citizenship and... Financial Support and Intent to Petition for Legal Custody for Public Law 97-359 Amerasian. (3) Agency form... ensure financial support for Public Law 97-359 Amerasian. The affidavit is used only to...

  4. 76 FR 41330 - Reports, Forms and Recordkeeping Requirements; Agency Information Collection Activity Under OMB...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-13

    ... ships for national defense and safety purposes. This collection consists of vessels that transmit their... automated collection techniques or other forms of information technology. A comment to OMB is best assured of having its full effect if OMB receives it within 30 days of publication. Authority: 49 CFR...

  5. 75 FR 65021 - Agency Information Collection Activities: Form I-485 and Supplements A and E, Extension of a...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-21

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form I-485 and... Security, U.S. Citizenship and Immigration Services (USCIS) will be submitting the following information..., and Supplements A and E; U.S. Citizenship and Immigration Services (USCIS). (4) Affected public...

  6. CTEPP DATA COLLECTION FORM 10 (PERIODS 1-3): DAY CARE CENTER CHILD ACTIVITY DIARY AND FOOD SURVEY

    EPA Science Inventory

    This data collection form collects information on the child's activities at the day care center over the 48-hr monitoring period. The diary is divided into three time periods over the 48-monitoring interval. The Food Survey collects information on the frequency and types of frui...

  7. High Energy Gamma Rays and Neutrinos from Star-forming Activities in the Galactic and Extragalactic Sources

    NASA Astrophysics Data System (ADS)

    Razzaque, Soebur

    2017-01-01

    The origin of the IceCube astrophysical neutrinos is an outstanding question. Star-forming activities which can accelerate particles to very high energies have been suggested as possible origin of these neutrinos. I will present a scenario where a subset of the neutrino events originate from the Galactic center region and Fermi Bubbles, resulting from star-forming activities. Multi-messenger signal in high energy gamma rays and neutrinos can probe this scenario. I will also present an analysis of the statistical association of the star-forming sources in our Galaxy and outside, with astrophysical neutrinos, as well as expected neutrino signal from these sources by fitting gamma-ray data.

  8. Anion- and pH-dependent activation of the soluble form of dopamine beta-hydroxylase.

    PubMed

    Terland, Ole; Flatmark, Torgeir

    2003-02-01

    The present study presents evidence for the conclusion that the catalytic activity of dopamine beta-hydroxylase (DBH; dopamine beta-mono-oxygenase, EC 1.14.17.1) is regulated independently by pH and by anions. In the absence of activating anions (i.e. in 50 mM Mes buffer) the activity was essentially zero at low pH (5.1-5.3) when assayed with the artificial electron donors ferrocyanide (0.25 mM), N, N, N', N'-tetramethyl-p-phenylenediamine (TMPD, 2 mM) or N, N-dimethyl-p-phenylenediamine (1 mM) and tyramine (8 mM) as the substrate to be hydroxylated. However, in the presence of activating anions (e.g. 0.05-0.6 M Cl(-) in 50 mM Mes buffer, 0.1 M phosphate buffer or 0.2 M acetate buffer) a high catalytic activity was observed at pH 5.1-5.3. The pronounced effect of anions at this pH may be related to the postulated anion-induced conformational change of DBH [Syvertsen, Melø and Ljones (1987) Biochim. Biophys. Acta 914, 6-18] resulting in a facilitated access of the substrates to the active site(s). The anion-activated DBH was inhibited when assayed with ferrocyanide and activated when assayed with TMPD as electron donors by increasing the pH (5.1 to 6.0). By contrast, in the absence of anions the activity increased from essentially zero at pH 5.1-5.3 to high values at pH 6.0, irrespective of the electron donor used. The data suggest that the conformational change induced by anion activation exposes a negatively charged group at or near the electron-donor-binding site(s) imposing an electrostatic repulsion towards ferrocyanide (four negative charges) and an electrostatic attraction towards the positively charged TMPD, thus explaining the different pH-activity curves obtained for the two electron donors. In contrast to the artificial electron donors, the physiological donor ascorbate [Terland and Flatmark (1975) FEBS Lett. 59, 52-56] supports hydroxylation of tyramine at low pH also in the absence of Cl(-), acetate or phosphate, confirming that ascorbate also

  9. Biologically active macromolecular forms of oxytocin. [8-Lysine]oxytocin as a suitable ligand.

    PubMed Central

    Snell, C R; Smyth, D G

    1977-01-01

    [8-Lysine]oxytocin was synthesized on a solid support and possessed an oxytocic activity of 100 +/- 6 units mumol on the isolated rat uterus. The epsilon-carbamoyl, epsilon-3-carboxypropionyl and epsilon-3-carboxybutryl derivatives were prepared and had uterotonic activities of 400, 55 and 50 units/mumol respectively. [8-Lysine]oxytocin was coupled unambiguously through the epsilon-amino group to the carboxyl groups of carboxymethylated dextrans or epsilon-3-carboxypropionly-gelatin. The macromolecular oxytocins were water-soluble and retained signigicant oxytocic activity. [8-Lysine]oxytocin should prove a useful ligand for affinity chromatography of oxytocin-binding proteins. PMID:889573

  10. 78 FR 77095 - Agency Information Collection Activities: Proposed Collection; Comment Request-Form FNS-648, WIC...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-20

    ... collection, the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) Local Agency..., Director, Supplemental Food Programs Division, Food and Nutrition Service, U.S. Department of Agriculture... Food and Nutrition Service Agency Information Collection Activities: Proposed Collection;...

  11. Closed-form expressions of some stochastic adapting equations for nonlinear adaptive activation function neurons.

    PubMed

    Fiori, Simone

    2003-12-01

    In recent work, we introduced nonlinear adaptive activation function (FAN) artificial neuron models, which learn their activation functions in an unsupervised way by information-theoretic adapting rules. We also applied networks of these neurons to some blind signal processing problems, such as independent component analysis and blind deconvolution. The aim of this letter is to study some fundamental aspects of FAN units' learning by investigating the properties of the associated learning differential equation systems.

  12. "Active surfaces" formed by immobilization of enzymes on solid-supported polymer membranes.

    PubMed

    Draghici, Camelia; Kowal, Justyna; Darjan, Alina; Meier, Wolfgang; Palivan, Cornelia G

    2014-10-07

    In various domains ranging from catalysis to medical and environmental sciences, there is currently much focus on the design of surfaces that present active compounds at the interface with their environments. Here, we describe the design of "active surfaces" based on solid-supported monolayers of asymmetric triblock copolymers, which serve as templates for the attachment of enzymes. A group of poly(ethylene glycol)-block-poly(γ-methyl-ε-caprolactone)-block-poly[(2-dimethylamino) ethyl methacrylate] amphiphilic copolymers, with different hydrophilic and hydrophobic domains (PEG45-b-PMCLx-b-PDMAEMAy) was selected to generate solid-supported polymer membranes. The behavior of the copolymers in terms of their molecular arrangements at the air-water interface was established by a combination of Langmuir isotherms and Brewster angle microscopy. Uniform thin layers of copolymers were obtained by transferring films onto silica solid supports at optimal surface pressure. These solid-supported polymer membranes were characterized by assessing various properties, such as monolayer thickness, hydrophilic/hydrophobic balance, topography, and roughness. Laccase, used as an enzyme model, was successfully attached to copolymer membranes by stable interactions as followed by quartz crystal microbalance with dissipation measurements, and its activity was preserved, as indicated by activity assays. The interaction between the amphiphilic triblock copolymer films and immobilized enzymes represents a straightforward approach to engineer "active surfaces", with biomolecules playing the active role by their intrinsic bioactivity.

  13. Elimination of Channel-Forming Activity by Insertional Inactivation of the p13 Gene in Borrelia burgdorferi

    PubMed Central

    Östberg, Yngve; Pinne, Marija; Benz, Roland; Rosa, Patricia; Bergström, Sven

    2002-01-01

    P13 is a chromosomally encoded 13-kDa integral outer membrane protein of the Lyme disease agent, Borrelia burgdorferi. The aim of this study was to investigate the function of the P13 protein. Here, we inactivated the p13 gene by targeted mutagenesis and investigated the porin activities of outer membrane proteins by using lipid bilayer experiments. Channel-forming activity was lost in the p13 mutant compared to wild-type B. burgdorferi, indicating that P13 may function as a porin. We purified native P13 to homogeneity by fast performance liquid chromatography and demonstrated that pure P13 has channel-forming activity with a single-channel conductance in 1 M KCl of 3.5 nS, the same as the porin activity that was lost in the p13 mutant. Further characterization of the channel formed by P13 suggested that it is cation selective and voltage independent. In addition, no major physiological effects of the inactivated p13 gene could be detected under normal growth conditions. The inactivation of p13 is the first reported inactivation of a gene encoding an integral outer membrane protein in B. burgdorferi. Here, we describe both genetic and biophysical experiments indicating that P13 in B. burgdorferi is an outer membrane protein with porin activity. PMID:12446631

  14. Polyclonal Anti-Peptide Antibodies Against Both Activated and Unactivated Forms of the Human Glucocorticoid Receptor

    DTIC Science & Technology

    1993-04-30

    1982, 198 7 ), using a variety of p r oteolytic enzymes , and insert i on and dele tion mutagenesis (Gigue:::-e et al . , 198~i Kumar et al ., 1986...synthetic peptide. To confirm the specificity of the antibody-GR interaction, antisera were preadsorbed with free peptide before reaction with the GR...immunization is a reproduc ible phenomenon. However, none of the three rabbits yielded high-titer serum for reaction with tne native forms of the hGR

  15. Additional Observations of Actively Forming Lava Tubes and Associated Structures, Hawaii

    NASA Technical Reports Server (NTRS)

    Greeley, Ronald

    1972-01-01

    Extensive changes occurred after the initial observations (Greeley, 1971) of lava tube and channel formation associated with the eruption of Mauna Ulu. Individual vents, which apparently acted somewhat independently, merged by collapse of intervening sections to form an elongate trench. Lava erupted from the summit vent flowed down the trench to the lower end and drained through lava tubes into Alae lava lake. Alae lava lake is in turn drained occasionally by other lava tubes and lava tube networks.

  16. Human embryonic stem cell-derived neuronal cells form spontaneously active neuronal networks in vitro.

    PubMed

    Heikkilä, Teemu J; Ylä-Outinen, Laura; Tanskanen, Jarno M A; Lappalainen, Riikka S; Skottman, Heli; Suuronen, Riitta; Mikkonen, Jarno E; Hyttinen, Jari A K; Narkilahti, Susanna

    2009-07-01

    The production of functional human embryonic stem cell (hESC)-derived neuronal cells is critical for the application of hESCs in treating neurodegenerative disorders. To study the potential functionality of hESC-derived neurons, we cultured and monitored the development of hESC-derived neuronal networks on microelectrode arrays. Immunocytochemical studies revealed that these networks were positive for the neuronal marker proteins beta-tubulin(III) and microtubule-associated protein 2 (MAP-2). The hESC-derived neuronal networks were spontaneously active and exhibited a multitude of electrical impulse firing patterns. Synchronous bursts of electrical activity similar to those reported for hippocampal neurons and rodent embryonic stem cell-derived neuronal networks were recorded from the differentiated cultures until up to 4 months. The dependence of the observed neuronal network activity on sodium ion channels was examined using tetrodotoxin (TTX). Antagonists for the glutamate receptors NMDA [D(-)-2-amino-5-phosphonopentanoic acid] and AMPA/kainate [6-cyano-7-nitroquinoxaline-2,3-dione], and for GABAA receptors [(-)-bicuculline methiodide] modulated the spontaneous electrical activity, indicating that pharmacologically susceptible neuronal networks with functional synapses had been generated. The findings indicate that hESC-derived neuronal cells can generate spontaneously active networks with synchronous communication in vitro, and are therefore suitable for use in developmental and drug screening studies, as well as for regenerative medicine.

  17. Hand-cycling: an active form of wheeled mobility, recreation, and sports.

    PubMed

    Hettinga, F J; Valent, L; Groen, W; van Drongelen, S; de Groot, S; van der Woude, L H V

    2010-02-01

    By studying exercise and performance in hand-cycling in both activities of daily living and in Paralympic sport settings, new insights can be gained for rehabilitation practice, adapted physical activity, and sports. This review looks into the pros and cons of hand-cycling in both rehabilitation and optimal sports performance settings as suggested from the current-but still limited-scientific literature and experimentation. Despite the limited evidence-base and the diversity of study approaches and methodologies, this study suggests an important role for hand-cycling during and after rehabilitation, and in wheeled mobility recreation and sports. An approach that combines biomechanical, physiological, and psychosocial elements may lead to a better understanding of the benefits of hand-cycling and of the fundamentals of exercise in rehabilitation, activities of daily living, and sports.

  18. Radiolabeling of intact dosage forms by neutron activation: effects on in vitro performance

    SciTech Connect

    Parr, A.; Jay, M.

    1987-12-01

    Compressed tablets containing various quantities of stable isotopes of Ba, Er, and Sm for use in neutron activation studies were evaluated for the effect of stable isotope incorporation on tablet hardness and disintegration times. At concentrations likely to be used in scintigraphic studies employing neutron activation as a radiolabeling method, no significant effect on in vitro parameters were observed. While the incorporation of stable isotopes influenced tablet hardness to a greater degree than disintegration time, irradiation of tablets in a neutron flux of 4.4 x 10(13) n/cm2 sec had a direct effect on tablet disintegration time. Thus, future neutron activation studies should focus on minimizing the amount of stable isotope to be incorporated with the formulation while using the shortest feasible irradiation time.

  19. Activation of Methane by the Pyridine Radical Cation and its Substituted Forms in the Gas Phase

    NASA Astrophysics Data System (ADS)

    Wu, Guohua; Stewart, Hamish; Liu, Zeyu; Wang, Yongcheng; Stace, Anthony J.

    2015-08-01

    We present an experimental study of methane activation by pyridine cation and its substituents in the gas phase. Mass spectrometric experiments in an ion trap demonstrate that pyridine cation and some of its substituent cations are able to react with methane. The deuterated methane experiment has confirmed that the hydrogen atom in the ionic product of reaction does come from methane. The collected information about kinetic isotope effects has been used to distinguish the nature of the bond activation as a hydrogen abstraction. Furthermore, experimental results demonstrated that the substituent groups on the pyridine ring can crucially influence their reactivity in methane bond activation processes. Density functional calculation (DFT) was employed to study the electronic structures of the complex and reaction mechanism of CH4+C5H5N+. The calculations confirmed the hypothesis from the experimental observation, namely, the reaction is rapid with no energy barrier.

  20. Gender differences in circulating endothelial progenitor cell colony-forming capacity and migratory activity in middle-aged adults.

    PubMed

    Hoetzer, Greta L; MacEneaney, Owen J; Irmiger, Heather M; Keith, Rebecca; Van Guilder, Gary P; Stauffer, Brian L; DeSouza, Christopher A

    2007-01-01

    Middle-aged women have a lower prevalence and incidence of cardiovascular events compared with men. The mechanisms responsible for this gender-specific difference are unclear. Numeric and functional impairments of bone marrow-derived circulating endothelial progenitor cells (EPCs) are associated with increased cardiovascular and cerebrovascular morbidity and mortality. It is currently unknown whether there are gender-related differences in EPC number and function in middle-aged adults. We tested the hypothesis that EPCs isolated from middle-aged women demonstrate greater colony-forming capacity and migratory activity compared with men of similar age. Peripheral blood samples were collected from 50 sedentary adults, 25 men (59 +/- 1 years of age) and 25 women (58 +/- 1 years of age). Mononuclear cells were isolated and preplated for 2 days, and nonadherent cells were further cultured for 7 days to determine EPC colony-forming units. Migratory activity of EPCs was determined using a modified Boyden chamber. The number of EPC colony-forming units was significantly higher (approximately 150%) in samples collected from women (16 +/- 3) compared with that collected from men (7 +/- 1). In addition, EPC migration (relative fluorescent units) was approximately 40% greater in women (729 +/- 74) than in men (530 +/- 67). In conclusion, these results demonstrate that EPC colony-forming capacity and migratory activity are higher in middle-aged women than in men.

  1. Pea formaldehyde-active class III alcohol dehydrogenase: common derivation of the plant and animal forms but not of the corresponding ethanol-active forms (classes I and P).

    PubMed Central

    Shafqat, J; El-Ahmad, M; Danielsson, O; Martínez, M C; Persson, B; Parés, X; Jornvall, H

    1996-01-01

    A plant class III alcohol dehydrogenase (or glutathione-dependent formaldehyde dehydrogenase) has been characterized. The enzyme is a typical class III member with enzymatic parameters and substrate specificity closely related to those of already established animal forms. Km values with the pea enzyme are 6.5 microM for NAD+, 2 microM for S-hydroxymethylglutathione, and 840 microM for octanol versus 9, 4, and 1200 microM, respectively, with the human enzyme. Structurally, the pea/human class III enzymes are closely related, exhibiting a residue identity of 69% and with only 3 of 23 residues differing among those often considered in substrate and coenzyme binding. In contrast, the corresponding ethanol-active enzymes, the long-known human liver and pea alcohol dehydrogenases, differ more (47% residue identities) and are also in functionally important active site segments, with 12 of the 23 positions exchanged, including no less than 7 at the usually much conserved coenzyme-binding segment. These differences affect functionally important residues that are often class-distinguishing, such as those at positions 48, 51, and 115, where the plant ethanol-active forms resemble class III (Thr, Tyr, and Arg, respectively) rather than the animal ethanol-active class I forms (typically Ser, His, and Asp, respectively). Calculations of phylogenetic trees support the conclusions from functional residues in subgrouping plant ethanol-active dehydrogenases and the animal ethanol-active enzymes (class I) as separate descendants from the class III line. It appears that the classical plant alcohol dehydrogenases (now called class P) have a duplicatory origin separate from that of the animal class I enzymes and therefore a paralogous relationship with functional convergence of their alcohol substrate specificity. Combined, the results establish the conserved nature of class III also in plants, and contribute to the molecular and functional understanding of alcohol dehydrogenases by

  2. Observations of actively forming lava tubes and associated structures, Hawaii, part 2

    NASA Technical Reports Server (NTRS)

    Greeley, R.

    1971-01-01

    A ground examination is made of lave tubes and channels. The surface morphology and the changes noted through lava flow activity are cited, and compared to earlier aerial observations. The lava activity was believed to be caused by a small lava lake exposed by the collapse of a crust covering it. Drainage of the lake was caused by a fissure erruption. New tubes or extensions of existing ones were noted from the flow. Molten lava was not seen in any tubes examined on the ground, but some of the flows were not sufficiently cooled to allow subsurface examination and survey of the tubes.

  3. Eravacycline (TP-434) Is Active In Vitro against Biofilms Formed by Uropathogenic Escherichia coli

    PubMed Central

    O'Brien, William; Kerstein, Kathryn O.; Sutcliffe, Joyce A.

    2015-01-01

    Eravacycline (formerly TP-434) was evaluated in vitro against pre-established biofilms formed by a uropathogenic Escherichia coli strain. Biofilms were eradicated by 0.5 μg/ml eravacycline, which was within 2-fold of the MIC for planktonic cells. In contrast, colistin and meropenem disrupted biofilms at 32 and 2 μg/ml, respectively, concentrations well above their respective MICs of 0.5 and 0.03 μg/ml. Gentamicin and levofloxacin eradicated biofilms at concentrations within 2-fold of their MICs. PMID:25624334

  4. Apoptotic activity of a nuclear form of mitogaligin, a cell death protein

    SciTech Connect

    Gonzalez, Patrick; Robinet, Pauline; Charpentier, Stephane; Mollet, Lucile; Normand, Thierry; Dubois, Martine; Legrand, Alain

    2009-01-23

    Galig, an internal gene to the galectin-3 gene, encodes two proteins and induces cell death in human cells. Mitogaligin, one of these proteins, contains a mitochondrial targeting sequence and promotes the release of cytochrome c into the cytosol. Here, we show that mitogaligin can also localize to nucleus. The nuclear form of mitogaligin induced cell death through a pathway exhibiting typical properties of apoptosis. These observations indicate for the first time that mitogaligin expresses cytotoxic properties not only when addressed to mitochondria but also when targeted to the nucleus.

  5. The Forming of Prospective Music Teacher's Readiness to Professional Activity in a Multicultural Society

    ERIC Educational Resources Information Center

    Salpykova, Indira M.; Politaeva, Tatyana I.

    2016-01-01

    The relevance of the researched problem is caused by the fact, that in modern social circumstances the special attention is given to the formation of a modern highly qualified music teacher, who should be prepared to implement his professional activity in a multicultural society, be able to treat the representatives of various social groups, their…

  6. 76 FR 64379 - Agency Information Collection Activities: Proposed Collection; Comments Requested; eForm 6 Access...

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  7. 77 FR 55486 - Agency Information Collection Activities: Employment Eligibility Verification, Form I-9, OMB...

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  8. 78 FR 63236 - Agency Information Collection Activities: Application To Replace Permanent Resident Card, Form...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-23

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Application To...-Day Notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration Services (USCIS) will be submitting the following information collection request to the Office...

  9. Hydroxynonenal inactivates cathepsin B by forming Michael adducts with active site residues.

    PubMed

    Crabb, John W; O'Neil, June; Miyagi, Masaru; West, Karen; Hoff, Henry F

    2002-04-01

    Oxidation of plasma low-density lipoprotein (oxLDL) generates the lipid peroxidation product 4-hydroxy-2 nonenal (HNE) and also reduces proteolytic degradation of oxLDL and other proteins internalized by mouse peritoneal macrophages in culture. This leads to accumulation of undegraded material in lysosomes and formation of ceroid, a component of foam cells in atherosclerotic lesions. To explore the possibility that HNE contributes directly to the inactivation of proteases, structure-function studies of the lysosomal protease cathepsin B have been pursued. We found that treatment of mouse macrophages with HNE reduces degradation of internalized maleyl bovine serine albumin and cathepsin B activity. Purified bovine cathepsin B treated briefly with 15 microM HNE lost approximately 76% of its protease activity and also developed immunoreactivity with antibodies to HNE adducts in Western blot analysis. After stabilization of the potential Michael adducts by sodium borohydride reduction, modified amino acids were localized within the bovine cathepsin B protein structure by mass spectrometric analysis of tryptic peptides. Michael adducts were identified by tandem mass spectrometry at cathepsin B active site residues Cys 29 (mature A chain) and His 150 (mature B chain). Thus, covalent interaction between HNE and critical active site residues inactivates cathepsin B. These results support the hypothesis that the accumulation of undegraded macromolecules in lysosomes after oxidative damage are caused in part by direct protease inactivation by adduct formation with lipid peroxidation products such as HNE.

  10. Function Follows Form: Activation of Shape and Function Features during Object Identification

    ERIC Educational Resources Information Center

    Yee, Eiling; Huffstetler, Stacy; Thompson-Schill, Sharon L.

    2011-01-01

    Most theories of semantic memory characterize knowledge of a given object as comprising a set of semantic features. But how does conceptual activation of these features proceed during object identification? We present the results of a pair of experiments that demonstrate that object recognition is a dynamically unfolding process in which function…

  11. 40 CFR 761.205 - Notification of PCB waste activity (EPA Form 7710-53).

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 32 2012-07-01 2012-07-01 false Notification of PCB waste activity... (CONTINUED) TOXIC SUBSTANCES CONTROL ACT POLYCHLORINATED BIPHENYLS (PCBs) MANUFACTURING, PROCESSING, DISTRIBUTION IN COMMERCE, AND USE PROHIBITIONS PCB Waste Disposal Records and Reports § 761.205 Notification...

  12. 40 CFR 761.205 - Notification of PCB waste activity (EPA Form 7710-53).

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 32 2013-07-01 2013-07-01 false Notification of PCB waste activity... (CONTINUED) TOXIC SUBSTANCES CONTROL ACT POLYCHLORINATED BIPHENYLS (PCBs) MANUFACTURING, PROCESSING, DISTRIBUTION IN COMMERCE, AND USE PROHIBITIONS PCB Waste Disposal Records and Reports § 761.205 Notification...

  13. 40 CFR 761.205 - Notification of PCB waste activity (EPA Form 7710-53).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Notification of PCB waste activity... (CONTINUED) TOXIC SUBSTANCES CONTROL ACT POLYCHLORINATED BIPHENYLS (PCBs) MANUFACTURING, PROCESSING, DISTRIBUTION IN COMMERCE, AND USE PROHIBITIONS PCB Waste Disposal Records and Reports § 761.205 Notification...

  14. An activated form of UFO alters leaf development and produces ectopic floral and inflorescence meristems.

    PubMed

    Risseeuw, Eddy; Venglat, Prakash; Xiang, Daoquan; Komendant, Kristina; Daskalchuk, Tim; Babic, Vivijan; Crosby, William; Datla, Raju

    2013-01-01

    Plants are unique in their ability to continuously produce new meristems and organ primordia. In Arabidopsis, the transcription factor LEAFY (LFY) functions as a master regulator of a gene network that is important for floral meristem and organ specification. UNUSUAL FLORAL ORGANS (UFO) is a co-activator of LEAFY and is required for proper activation of APETALA3 in the floral meristem during the specification of stamens and petals. The ufo mutants display defects in other parts of the flower and the inflorescence, suggestive of additional roles. Here we show that the normal determinacy of the developing Arabidopsis leaves is affected by the expression of a gain-of-function UFO fusion protein with the VP16 transcriptional activator domain. In these lines, the rosette and cauline leaf primordia exhibit reiterated serration, and upon flowering produce ectopic meristems that develop into flowers, bract leaves and inflorescences. These striking phenotypes reveal that developing leaves maintain the competency to initiate flower and inflorescence programs. Furthermore, the gain-of-function phenotypes are dependent on LFY and the SEPALLATA (SEP) MADS-box transcription factors, indicative of their functional interactions with UFO. The findings of this study also suggest that UFO promotes the establishment of the lateral meristems and primordia in the peripheral zone of the apical and floral meristems by enhancing the activity of LFY. These novel phenotypes along with the mutant phenotypes of UFO orthologs in other plant species suggest a broader function for UFO in plants.

  15. Phosphorylation and activation of a transducible recombinant form of human HSP20 in E. coli

    PubMed Central

    Flynn, Charles R.; Smoke, Christopher C.; Furnish, Elizabeth; Komalavilas, Padmini; Thresher, Jeffrey; Yi, Zhengping; Mandarino, Lawrence J.; Brophy, Colleen M.

    2007-01-01

    Protein based cellular therapeutics have been limited by getting the molecules into cells and the fact that many proteins require post-translational modifications for activation. Protein transduction domains (PTDs), including that from the HIV TAT protein (TAT), are small arginine rich peptides that carry molecules across the cell membrane. We have shown that the heat shock-related protein, HSP20 is a downstream mediator of cyclic nucleotide-dependent relaxation of vascular smooth muscle and is activated by phosphorylation. In this study, we co-expressed in E. coli the cDNAs encoding the catalytic subunit of protein kinase G and a TAT-HSP20 fusion protein composed of the TAT PTD (-YGRKKRRQRRR-) fused to the N-terminus of human HSP20. Immunoblot and HPLC-ESI-MS/MS analysis of the purified TAT-HSP20 demonstrated that it was phosphorylated at serine 40 (equivalent to serine 16 in wild-type human HSP20). This phosphorylated TAT-HSP20 was physiologically active in intact smooth muscles in that it inhibited 5-hydroxytryptamine-induced contractions by 57% ± 4.5. The recombinant phosphorylated protein also led to changes in actin cytoskeletal morphology in 3T3 cells. These results delineate strategies for the expression and activation of therapeutic molecules for intracellular protein based therapeutics. PMID:17084643

  16. 78 FR 62644 - Agency Information Collection Activities: Application for Temporary Protected Status, Form I-821...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-22

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  17. 78 FR 63487 - Agency Information Collection Activities: Petition for a Nonimmigrant Worker, Form I-129...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-24

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  18. 78 FR 35639 - Agency Information Collection Activities: Application To Extend/Change Nonimmigrant Status, Form...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-13

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Application To...-Day Notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration... utility; (2) Evaluate the accuracy of the agency's estimate of the burden of the proposed collection...

  19. 78 FR 44141 - Agency Information Collection Activities: Affidavit of Support, Form I-134; Extension, Without...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-23

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Affidavit of.... SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration Services (USCIS... accuracy of the agency's estimate of the burden of the proposed collection of information, including...

  20. An Activated Form of UFO Alters Leaf Development and Produces Ectopic Floral and Inflorescence Meristems

    PubMed Central

    Risseeuw, Eddy; Venglat, Prakash; Xiang, Daoquan; Komendant, Kristina; Daskalchuk, Tim; Babic, Vivijan; Crosby, William; Datla, Raju

    2013-01-01

    Plants are unique in their ability to continuously produce new meristems and organ primordia. In Arabidopsis, the transcription factor LEAFY (LFY) functions as a master regulator of a gene network that is important for floral meristem and organ specification. UNUSUAL FLORAL ORGANS (UFO) is a co-activator of LEAFY and is required for proper activation of APETALA3 in the floral meristem during the specification of stamens and petals. The ufo mutants display defects in other parts of the flower and the inflorescence, suggestive of additional roles. Here we show that the normal determinacy of the developing Arabidopsis leaves is affected by the expression of a gain-of-function UFO fusion protein with the VP16 transcriptional activator domain. In these lines, the rosette and cauline leaf primordia exhibit reiterated serration, and upon flowering produce ectopic meristems that develop into flowers, bract leaves and inflorescences. These striking phenotypes reveal that developing leaves maintain the competency to initiate flower and inflorescence programs. Furthermore, the gain-of-function phenotypes are dependent on LFY and the SEPALLATA (SEP) MADS-box transcription factors, indicative of their functional interactions with UFO. The findings of this study also suggest that UFO promotes the establishment of the lateral meristems and primordia in the peripheral zone of the apical and floral meristems by enhancing the activity of LFY. These novel phenotypes along with the mutant phenotypes of UFO orthologs in other plant species suggest a broader function for UFO in plants. PMID:24376756

  1. 75 FR 19630 - Commission Information Collection Activities (FERC Form 80); Comment Request; Extension

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-15

    ... will send an automatic acknowledgement to the sender's e-mail address upon receipt of eFiled comments... receiving automatic notification of activity in this docket may do so through eSubscription at http://www..., printed or downloaded remotely through FERC's eLibrary at:...

  2. 78 FR 1218 - Agency Information Collection Activities: Nonimmigrant Petition Based on Blanket L Petition; Form...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-08

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Nonimmigrant...-Day Notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration..., Regulatory Coordination Division, Office of Policy and Strategy, U.S. Citizenship and Immigration...

  3. Effect of human serum albumin upon the permeabilizing activity of sticholysin II, a pore forming toxin from Stichodactyla heliantus.

    PubMed

    Celedón, Gloria; González, Gustavo; Gulppi, Felipe; Pazos, Fabiola; Lanio, María E; Alvarez, Carlos; Calderón, Cristian; Montecinos, Rodrigo; Lissi, Eduardo

    2013-12-01

    Sticholysin II (St II) is a haemolytic toxin isolated from the sea anemone Stichodactyla helianthus. The high haemolytic activity of this toxin is strongly dependent on the red cell status and the macromolecule conformation. In the present communication we evaluate the effect of human serum albumin on St II haemolytic activity and its capacity to form pores in the bilayer of synthetic liposomes. St II retains its pore forming capacity in the presence of large concentrations (up to 500 μM) of human serum albumin. This effect is observed both in its capacity to produce red blood cells haemolysis and to generate functional pores in liposomes. In particular, the capacity of the toxin to lyse red blood cells increases in the presence of human serum albumin (HSA). Regarding the rate of the pore forming process, it is moderately decreased in liposomes and in red blood cells, in spite of an almost total coverage of the interface by albumin. All the data obtained in red cells and model membranes show that St II remains lytically active even in the presence of high HSA concentrations. This stubbornness can explain why the toxin is able to exert its haemolytic activity on membranes immersed in complex plasma matrixes such as those present in living organisms.

  4. Action of a cationic surfactant on the activity and removal of bacterial biofilms formed under different flow regimes.

    PubMed

    Simões, Manuel; Pereira, Maria Olivia; Vieira, Maria João

    2005-01-01

    The action of the cationic surfactant cetyltrimethylammonium bromide (CTAB) was investigated to control biofilms (aged 7d) formed by Pseudomonas fluorescens on stainless-steel slides, using flow cells reactors, under turbulent and laminar flow. The effect of CTAB was also investigated using planktonic cells in the presence and absence of BSA, by measuring the cellular respiratory activity and the ATP released. The action of CTAB on biofilms was assessed by means of cellular respiratory activity and variation of biofilm mass, immediately and 3, 7 and 12h after the application of CTAB. The physical stability of the biofilm was also assessed using a rotating device, where the effect of the surfactant on the biofilm stability was evaluated through the variation of the mass remaining on the surface. CTAB significantly reduced the activity of the planktonic cells probably due to the rupture of the cells. This effect was significantly reduced in the presence of BSA. Planktonic cells were more easily inactivated than bacteria in biofilms. Biofilms formed under laminar flow were more susceptible than those formed under turbulent flow, but in both cases total inactivation was not achieved. Biofilm recovery was observed, in terms of respiratory activity, in almost all the cases studied. CTAB application by itself did not promote the detachment of biofilms. The physical stability tests showed that the synergistic action of the surfactant and the application of high shear stress to the biofilm increase its detachment.

  5. Pharmacotechnical caracterization and effectiveness study of a dermopharmaceutical form: rosemary honey contributions as a moisturizing active.

    PubMed

    Jiménez Soriano, M M; Fresno Contreras, M J; Sellés Flores, E

    1999-09-01

    We have designed, elaborated and studied a dermopharmaceutical form formulated on the basis of a modern self-emulsifying excipient and rosemary honey (known as Miel de La Alcarria--Spain--according to the Governing Council), in order to obtain a hight degree of cutaneous hydratation. The formulation is typified and characterized from a pharmacotechnical and rheological points of view. In this sense, the experimental protocol has emphasized rheological essays which give relevant practical information. Also, we have performed a complete study of it physical and structural stability, and, lastly, we evaluated the dermopharmaceutical effectiveness. The work plan included the following tests: 1) Pharmacotechnical Essays--organoleptic characteristics, photomicrograh study, type of interposition, pH-determination, rheological and thixotropic study and physical stability tests; 2) Dermopharmaceutical Effectiveness Assays--Corneometric and Sebumetric measurements. From the results, we have deduced that the emulsified binary system that is proposed, stable from a physical and structural points of view, presents confirmed properties and a very good cosmetological adequation. In this sense, our emulsion presents a high degree of moisturizing/emollient power that qualifies it not only as a magnificent eudermic dermopharmaceutical form, but also as a very appropriate vehicle for Dermopharmaceutical and/or Dermatological Formulation.

  6. Pore-forming Activity of the Escherichia coli Type III Secretion System Protein EspD*

    PubMed Central

    Chatterjee, Abhishek; Caballero-Franco, Celia; Bakker, Dannika; Totten, Stephanie; Jardim, Armando

    2015-01-01

    Enterohemorrhagic Escherichia coli is a causative agent of gastrointestinal and diarrheal diseases. Pathogenesis associated with enterohemorrhagic E. coli involves direct delivery of virulence factors from the bacteria into epithelial cell cytosol via a syringe-like organelle known as the type III secretion system. The type III secretion system protein EspD is a critical factor required for formation of a translocation pore on the host cell membrane. Here, we show that recombinant EspD spontaneously integrates into large unilamellar vesicle (LUV) lipid bilayers; however, pore formation required incorporation of anionic phospholipids such as phosphatidylserine and an acidic pH. Leakage assays performed with fluorescent dextrans confirmed that EspD formed a structure with an inner diameter of ∼2.5 nm. Protease mapping indicated that the two transmembrane helical hairpin of EspD penetrated the lipid layer positioning the N- and C-terminal domains on the extralumenal surface of LUVs. Finally, a combination of glutaraldehyde cross-linking and rate zonal centrifugation suggested that EspD in LUV membranes forms an ∼280–320-kDa oligomeric structure consisting of ∼6–7 subunits. PMID:26324713

  7. Calcium Oxalate Stone Agglomeration Inhibition [tm] Reflects Renal Stone-Forming Activity.

    PubMed

    Lindberg, J S; Cole, F E; Romani, W; Husserl, F E; Fuselier, H A; Kok, D J; Erwin, D T

    2000-04-01

    Louisiana and other Gulf South states comprise a "Stone Belt" where calcium oxalate stone formers (CaOx SFs) are found at a high rate of approximately 5%. In these patients, the agglomeration of small stone crystals, which are visible in nearly all morning urine collections, forms stones that can become trapped in the renal parenchyma and the renal pelvis. Without therapy, about half of CaOx SFs repeatedly form kidney stones, which can cause excruciating pain that can be relieved by passage, fragmentation (lithotripsy), or surgical removal. The absence of stones in "normal" patients suggests that there are stone inhibitors in "normal" urines.At the Ochsner Renal Stone Clinic, 24-hour urine samples are collected by the patient and sent to the Ochsner Renal Stone Research Program where calcium oxalate stone agglomeration inhibition [tm] measurements are performed. Urine from healthy subjects and inactive stone formers has demonstrated strongly inhibited stone growth [tm] in contrast to urine from recurrent CaOx SFs. [tm] data from 1500 visits of 700 kidney stone patients have been used to evaluate the risk of recurrence in Ochsner's CaOx SF patients. These data have also been used to demonstrate the interactive roles of certain identified urinary stone-growth inhibitors, citrate and Tamm-Horsfall protein (THP), which can be manipulated with medication to diminish recurrent stone formation. Our goal is to offer patients both financial and pain relief by reducing their stones with optimized medication, using medical management to avoid costly treatments.

  8. Antioxidant activity evaluation of new dosage forms as vehicles for dehydrated vegetables.

    PubMed

    Romero-de Soto, María Dolores; García-Salas, Patricia; Fernández-Arroyo, Salvador; Segura-Carretero, Antonio; Fernández-Campos, Francisco; Clares-Naveros, Beatriz

    2013-06-01

    A dehydrated vegetables mixture loaded in four pharmaceutical dosage forms as powder, effervescent granulate, sugar granulate and gumdrops were investigated for their antioxidant capacity using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging capacity assay, oxygen radical absorbance capacity assay and ferric reducing antioxidant potential assay. Total phenolic content of dehydrated vegetables powder mixture was also measured by the Folin-Ciocalteu method, so as to evaluate its contribution to their total antioxidant function. The effect of different temperatures on stability of these systems after 90 days storage was also evaluated. These formulations presented strong antioxidant properties and high phenolic content (279 mg gallic acid equivalent/g of sample) and thus could be potential rich sources of natural antioxidants. Antioxidant properties differed significantly among selected formulations (p < 0.05). Generally, the losses were lower in samples stored under refrigeration. To interpret the antioxidant properties a kinetic approach was performed. Degradation kinetics for the phenolic content and antioxidant capacity followed a zero-order function. Effervescent granulate was the formulation which underwent faster degradation. Contrary, sugar granulate and gumdrops were much more slowly. Time required to halve the initial amount of phenolic compounds was 589 ± 45 days for samples stored at 4 º C, and 312 ± 16 days for samples stored at room temperature. These developed dosage forms are new and innovative approach for vegetable intakes in population with special requirements providing an improvement in the administration of vegetables and fruits.

  9. Five of 12 forms of vaccinia virus-expressed human hepatic cytochrome P450 metabolically activate aflatoxin B sub 1

    SciTech Connect

    Aoyama, Toshifumi; Yamano, Shigeru; Gelboin, H.V.; Gonzalez, F.J. ); Guzelian, P.S. )

    1990-06-01

    Twelve forms of human hepatic cytochrome P450 were expressed in hepatoma cells by means of recombinant vaccinia viruses. The expressed P450s were analyzed for their abilities to activate the potent hepatocarcinogen aflatoxin B{sub 1} to metabolites having mutagenic or DNA-binding properties. Five forms, P450s IA2, IIA3, IIB7, IIIA3, and IIIA4, activated aflatoxin B{sub 1} to mutagenic metabolites as assessed by the production of His revertants of Salmonella typhimurium in the Ames test. The same P450s catalyzed conversion of aflatoxin B{sub 1} to DNA-bound derivatives as judged by an in situ assay in which the radiolabeled carcinogen was incubated with cells expressing the individual P450 forms. Seven other human P450s, IIC8, IIC9, IID6, IIE1, IIF1, and IIIA5, and IVB1, did not significantly activate aflatoxin B{sub 1} as measured by both the Ames test and the DNA-binding assay. Moreover, polyclonal anti-rat liver P450 antibodies that crossreact with individual human P450s IA2, IIA3, IIIA3, and IIIA4 each inhibited aflatoxin B{sub 1} activation catalyzed by human liver S-9 extracts. Inhibition ranged from as low as 10% with antibody against IIA3 to as high as 65% with antibody against IIIA3 and IIIA4. These results establish that metabolic activation of aflatoxin B{sub 1} in human liver involves the contribution of multiple forms of P450.

  10. Early Activation of STAT3 Regulates Reactive Astrogliosis Induced by Diverse Forms of Neurotoxicity

    PubMed Central

    O'Callaghan, James P.; Kelly, Kimberly A.; VanGilder, Reyna L.; Sofroniew, Michael V.; Miller, Diane B.

    2014-01-01

    Astrogliosis, a cellular response characterized by astrocytic hypertrophy and accumulation of GFAP, is a hallmark of all types of central nervous system (CNS) injuries. Potential signaling mechanisms driving the conversion of astrocytes into “reactive” phenotypes differ with respect to the injury models employed and can be complicated by factors such as disruption of the blood-brain barrier (BBB). As denervation tools, neurotoxicants have the advantage of selective targeting of brain regions and cell types, often with sparing of the BBB. Previously, we found that neuroinflammation and activation of the JAK2-STAT3 pathway in astrocytes precedes up regulation of GFAP in the MPTP mouse model of dopaminergic neurotoxicity. Here we show that multiple mechanistically distinct mouse models of neurotoxicity (MPTP, AMP, METH, MDA, MDMA, KA, TMT) engender the same neuroinflammatory and STAT3 activation responses in specific regions of the brain targeted by each neurotoxicant. The STAT3 effects seen for TMT in the mouse could be generalized to the rat, demonstrating cross-species validity for STAT3 activation. Pharmacological antagonists of the neurotoxic effects blocked neuroinflammatory responses, pSTAT3tyr705 and GFAP induction, indicating that damage to neuronal targets instigated astrogliosis. Selective deletion of STAT3 from astrocytes in STAT3 conditional knockout mice markedly attenuated MPTP-induced astrogliosis. Monitoring STAT3 translocation in GFAP-positive cells indicated that effects of MPTP, METH and KA on pSTAT3tyr705 were localized to astrocytes. These findings strongly implicate the STAT3 pathway in astrocytes as a broadly triggered signaling pathway for astrogliosis. We also observed, however, that the acute neuroinflammatory response to the known inflammogen, LPS, can activate STAT3 in CNS tissue without inducing classical signs of astrogliosis. Thus, acute phase neuroinflammatory responses and neurotoxicity-induced astrogliosis both signal through

  11. Electrocatalytically Active Nickel-Based Electrode Coatings Formed by Atmospheric and Suspension Plasma Spraying

    NASA Astrophysics Data System (ADS)

    Aghasibeig, M.; Mousavi, M.; Ben Ettouill, F.; Moreau, C.; Wuthrich, R.; Dolatabadi, A.

    2014-01-01

    Ni-based electrode coatings with enhanced surface areas, for hydrogen production, were developed using atmospheric plasma spray (APS) and suspension plasma spray (SPS) processes. The results revealed a larger electrochemical active surface area for the coatings produced by SPS compared to those produced by APS process. SEM micrographs showed that the surface microstructure of the sample with the largest surface area was composed of a large number of small cauliflower-like aggregates with an average diameter of 10 μm.

  12. Postnatal Chick Choroids Exhibit Increased Retinaldehyde Dehydrogenase Activity During Recovery From Form Deprivation Induced Myopia

    PubMed Central

    Harper, Angelica R.; Wang, Xiang; Moiseyev, Gennadiy; Ma, Jian-Xing; Summers, Jody A.

    2016-01-01

    Purpose Increases in retinaldehyde dehydrogenase 2 (RALDH2) transcript in the chick choroid suggest that RALDH2 may be responsible for increases observed in all-trans-retinoic acid (atRA) synthesis during recovery from myopic defocus. The purpose of the present study was to examine RALDH2 protein expression, RALDH activity, and distribution of RALDH2 cells in control and recovering chick ocular tissues. Methods Myopia was induced in White Leghorn chicks for 10 days, followed by up to 15 days of unrestricted vision (recovery). Expression of RALDH isoforms in chick ocular tissues was evaluated by Western blot. Catalytic activity of RALDH was measured in choroidal cytosol fractions using an in vitro atRA synthesis assay together with HPLC quantification of synthesized atRA. Distribution of RALDH2 cells throughout the choroid was evaluated by immunohistochemistry. Results RALDH2 was expressed predominately in the chick choroid (P < 0.001) and increased after 24 hours and 4 days of recovery (76%, 74%, and 165%, respectively; P < 0.05). Activity of RALDH was detected solely in the choroid and was elevated at 3 and 7 days of recovery compared to controls (70% and 48%, respectively; P < 0.05). The number of RALDH2 immunopositive cells in recovering choroids was increased at 24 hours and 4 to 15 days of recovery (P < 0.05) and were concentrated toward the RPE side compared to controls. Conclusions The results of this study suggest that RALDH2 is the major RALDH isoform in the chick choroid and is responsible for the increased RALDH activity seen during recovery. PMID:27654415

  13. Nitroxide polymer networks formed by Michael addition: on site-cured electrode-active organic coating.

    PubMed

    Ibe, Takeshi; Frings, Rainer B; Lachowicz, Artur; Kyo, Soichi; Nishide, Hiroyuki

    2010-05-28

    Highly and homogeneously crosslinked poly(beta-ketoester) networks densely bearing robust nitroxide radicals were prepared via a click-type and stepwise Michael polyaddition. A half-battery cell composed of the thermally-cured radical network coatings displayed a rapid, reversible, and almost stoichiometric redox-activity even with a thickness of ca. 10 mum, which may be applicable as the electrode of organic-based rechargeable devices.

  14. Voltammetric determination of ruthenium in the form of complexes with biologically active ligands

    SciTech Connect

    Medyantseva, E.P.; Budnikov, G.K.; Balakaeva, T.A.

    1992-02-10

    The interest in the analytical chemistry of ruthenium and its compounds has recently been increasing. Ruthenium compounds can be used an antitumor agents and in the treatment of tuberculosis and fungal infections. It has been suggested that there is a specific relationship between the reduction potentials of the compounds and their biological activity. Of greatest interest among the biologically active compounds are the compounds with nitrogen-containing heterocycles. In order to obtain information on the degree of oxidation of the central atom in the complexes and to select the optimum conditions for the determination of the mono- and bi-nuclear complexes of ruthenium compounds with biologically active ligands such as imidazole (Im), histidine (His), benzimidazole (BIm) and its methyl derivative [1,2(CH{sub 3}){sub 2} - BIm], benzohyroxamic acid (Bha), and 1-phenyl-2-methylamino-1-propanol or ephedrine (Eph) in the present work, the authors studied their electrochemical behavior at dropping mercury (dme) and a platinum electrodes. 6 refs., 1 fig., 2 tabs.

  15. Inhibition of poly(adenosine diphosphate-ribose) polymerase by the active form of vitamin D

    PubMed Central

    MABLEY, JON G.; WALLACE, REBECCA; PACHER, PÁL; MURPHY, KANNEGANTI; SZABÓ, CSABA

    2008-01-01

    Vitamin D is well characterized for its role in mineral homeostasis and maintenance of normal skeletal architecture. Vitamin D has been demonstrated to exert anti-inflammatory effects in a variety of disease states including diabetes, arthritis and inflammatory bowel disease. In these diseases poly[adenosine diphosphate (ADP)-ribose] polymerase (PARP) inhibitors have also proved effective as anti-inflammatory agents. Here we present data demonstrating that the active metabolite of vitamin D, 1α,25-dihydroxyvitamin D3, is a PARP inhibitor. UV irradiation-mediated PARP activation in human keratinocytes can be inhibited by treatment with vitamin D, 7-dehydrocholesterol or 1α,25-dihydroxyvitamin D3. Inhibition of cytochrome P450 reversed the PARP inhibitory action of vitamin D and 7-dehydrocholesterol, indicating that conversion to 1α,25-dihydroxyvitamin D3 mediates their PARP inhibitory action. Vitamin D may protect keratinocytes against over-activation of PARP resulting from exposure to sunlight. PARP inhibition may contribute to the pharmacological and anti-inflammatory effects of vitamin D. PMID:17487428

  16. Altered nucleosomes of active nucleolar chromatin contain accessible histone H3 in its hyperacetylated forms

    SciTech Connect

    Johnson, E.M.; Sterner, R.; Allfrey, V.G.

    1987-05-25

    Chromatin of the organism Physarum polycephalum contains a class of conformationally altered nucleosomes previously localized to the transcribing regions of ribosomal genes in nucleoli. When nuclei are treated with 2-iodo(2-tritium)acetate, the histone H3 sulfhydryl group of the altered nucleosomes is derivatized while that of folded nucleosomes is not, and the labeled histones can then be identified by autoradiography of gels that separate H3 isoforms. The H3 derivatized is predominantly of tri- and tetraacetylated forms. In contrast, total free histone reacted with iodoacetate shows no preferential labeling of isoforms. Selective reaction of acetylated H3 is prevalent in both nucleolar and non-nucleolar chromatin. The results link specific patterns of H3 acetylation to changes in nucleosome conformation that occur during transcription.

  17. The role of sulfatide lipid domains in the membrane pore-forming activity of cobra cardiotoxin.

    PubMed

    Wu, Po-Long; Chiu, Chang-Ru; Huang, Wei-Ning; Wu, Wen-Guey

    2012-05-01

    Cobra CTX A3, the major cardiotoxin (CTX) from Naja atra, is a cytotoxic, basic β-sheet polypeptide that is known to induce a transient membrane leakage of cardiomyocytes through a sulfatide-dependent CTX membrane pore formation and internalization mechanism. The molecular specificity of CTX A3-sulfatide interaction at atomic levels has also been shown by both nuclear magnetic resonance (NMR) and X-ray diffraction techniques to reveal a role of CTX-induced sulfatide conformational changes for CTX A3 binding and dimer formation. In this study, we investigate the role of sulfatide lipid domains in CTX pore formation by various biophysical methods, including fluorescence imaging and atomic force microscopy, and suggest an important role of liquid-disordered (ld) and solid-ordered (so) phase boundary in lipid domains to facilitate the process. Fluorescence spectroscopic studies on the kinetics of membrane leakage and CTX oligomerization further reveal that, although most CTXs can oligomerize on membranes, only a small fraction of CTXs oligomerizations form leakage pores. We therefore suggest that CTX binding at the boundary between the so and so/ld phase coexistence sulfatide lipid domains could form effective pores to significantly enhance the CTX-induced membrane leakage of sulfatide-containing phosphatidylcholine vesicles. The model is consistent with our earlier observations that CTX may penetrate and lyse the bilayers into small aggregates at a lipid/protein molar ratio of about 20 in the ripple P(β)' phase of phosphatidylcholine bilayers and suggest a novel mechanism for the synergistic action of cobra secretary phospholipase A2 and CTXs.

  18. Permission Forms

    ERIC Educational Resources Information Center

    Zirkel, Perry A.

    2005-01-01

    The prevailing practice in public schools is to routinely require permission or release forms for field trips and other activities that pose potential for liability. The legal status of such forms varies, but they are generally considered to be neither rock-solid protection nor legally valueless in terms of immunity. The following case and the…

  19. Archaeal orthologs of Cdc45 and GINS form a stable complex that stimulates the helicase activity of MCM

    PubMed Central

    Xu, Yuli; Gristwood, Tamzin; Hodgson, Ben; Trinidad, Jonathan C.; Albers, Sonja-Verena; Bell, Stephen D.

    2016-01-01

    The regulated recruitment of Cdc45 and GINS is key to activating the eukaryotic MCM(2-7) replicative helicase. We demonstrate that the homohexameric archaeal MCM helicase associates with orthologs of GINS and Cdc45 in vivo and in vitro. Association of these factors with MCM robustly stimulates the MCM helicase activity. In contrast to the situation in eukaryotes, archaeal Cdc45 and GINS form an extremely stable complex before binding MCM. Further, the archaeal GINS•Cdc45 complex contains two copies of Cdc45. Our analyses give insight into the function and evolution of the conserved core of the archaeal/eukaryotic replisome. PMID:27821767

  20. Arf1 and Arf6 Promote Ventral Actin Structures formed by acute Activation of Protein Kinase C and Src

    PubMed Central

    Caviston, Juliane P.; Cohen, Lee Ann; Donaldson, Julie G.

    2016-01-01

    Arf proteins regulate membrane traffic and organelle structure. Although Arf6 is known to initiate actin-based changes in cell surface architecture, Arf1 may also function at the plasma membrane. Here we show that acute activation of protein kinase C (PKC) induced by the phorbol ester PMA led to the formation of motile actin structures on the ventral surface of Beas-2b cells, a lung bronchial epithelial cell line. Ventral actin structures also formed in PMA-treated HeLa cells that had elevated levels of Arf activation. For both cell types, formation of the ventral actin structures was enhanced by expression of active forms of either Arf1 or Arf6, and by the expression of guanine nucleotide exchange factors that activate these Arfs. By contrast, formation of these structures was blocked by inhibitors of PKC and Src, and required phosphatidylinositol 4, 5-bisphosphate, Rac, Arf6 and Arf1. Furthermore, expression of ASAP1, an Arf1 GTPase activating protein (GAP) was more effective at inhibiting the ventral actin structures than was ACAP1, an Arf6 GAP. This study adds to the expanding role for Arf1 in the periphery and identifies a requirement for Arf1, a “Golgi Arf”, in the reorganization of the cortical actin cytoskeleton on ventral surfaces, against the substratum. PMID:24916416

  1. Investigation of Channel-Forming Activity of Polyene Macrolide Antibiotics in Planar Lipid Bilayers in the Presence of Dipole Modifiers

    PubMed Central

    Efimova, S. S.; Schagina, L. V.; Ostroumova, O. S.

    2014-01-01

    The role of membrane components, sterols, phospholipids and sphingolipids in the formation and functioning of ion-permeable nanopores formed by antifungal macrolide antibiotics, amphotericin B, nystatin and filipin in planar lipid bilayers was studied. Dipole modifiers, flavonoids and styryl dyes, were used as a tool to study the molecular mechanisms of polyene channel-forming activity. The introduction of dipole modifiers into the membrane bathing solutions was shown to change the conductance of single channels and the steadystate transmembrane current induced by polyene antibiotics in the sterol-containing phospholipid-bilayers. The conductance of single amphotericin B channels was found to depend on the dipole potential of the membrane. The experiments with various phospholipids, sterols, and polyenes led to the assumption that the shape of a phospholipid molecule, the presence of double bonds at the positions 7 and 22 of a sterol molecule, the number of conjugated double bonds, and the presence of an amino sugar in the polyene antibiotic molecule are important factors impacting the stability of polyene-lipid complexes forming ion-permeable pores. Experimental and literature data presented in the paper suggest that the channel-forming activity of polyene antibiotics is also affected by the physicochemical properties of polyene-enriched ordered membrane domains. PMID:25558397

  2. A monoclonal antibody that distinguishes latent and active forms of the proteasome (multicatalytic proteinase complex)

    NASA Technical Reports Server (NTRS)

    Weitman, D.; Etlinger, J. D.

    1992-01-01

    Monoclonal antibodies (mAbs) were generated to proteasome purified from human erythrocytes. Five of six proteasome-specific mAbs reacted with three subunits in the molecular mass range of 25-28 kDa, indicating a common epitope. The other mAb (AP5C10) exhibited a more restricted reactivity, recognizing a 32-kDa subunit of the proteasome purified in its latent state. However, when the proteasome is isolated in its active state, AP5C10 reacts with a 28-kDa subunit, evidence for processing of the proteasome subunits during purification. Purified proteasome preparations which exhibited partial latency have both AP5C10 reactive subunits. Although the 32-kDa subunit appears required for latency, loss of this component and generation of the 28-kDa component are not obligatory for activation. The 32- and 28-kDa subunits can each be further resolved into three components by isoelectric focusing. The apparent loss of 4 kDa during the conversion of the 32- to 28-kDa subunit is accompanied by a shift to a more basic pI for each polypeptide. Western blots of the early steps of proteasome purification reveal an AP5C10-reactive protein at 41 kDa. This protein was separated from proteasomes by sizing chromatography and may represent a pool of precursor subunits. Since the 32-kDa subunit appears necessary for latency, it is speculated to play a regulatory role in ATP-dependent proteolytic activity.

  3. Activity of MKT 077, a rhodacyanine dye, against human tumor colony-forming units.

    PubMed

    Petit, T; Izbicka, E; Lawrence, R A; Nalin, C; Weitman, S D; Von Hoff, D D

    1999-03-01

    MKT 077 is related to rhodamine 123 dye and demonstrates preferential accumulation in the mitochondria of cancer cells compared to normal cells. This difference in retention between cancer and normal cells led to the finding that MKT 077 selectively inhibits the growth of cancer cells in vitro. To define the preclinical activity profile of MKT 077, the compound was tested in vivo against a large variety of human tumors utilizing the human tumor-cloning assay. MKT 077 was studied using a sequential 2 h exposure separated by 24 h (2-24-2 h) and a 24 h exposure at final concentrations of 0.1, 0.2, 1.0, 2.0, 10.0 and 20.0 microg/ml. MKT 077 was also studied using continuous exposure at final concentrations of 0.1, 1.0 and 10 microg/ml. A decrease in tumor colony formation was considered significant if survival of colonies treated with MKT 077 was 50% or less compared to untreated controls. A total of 149 specimens was treated with MKT 077 with 51, 58 and 34 evaluable specimens with the 2-24-2 h, the 24 h and the continuous exposure, respectively. The results of the present study suggest a positive relationship between concentration and response. No relationship between exposure schedule and activity was observed. Inhibitory effects were obtained against multiple tumor types. High cytotoxic activity was obtained against breast, ovary, endometrial, colon and non-small cell lung cancer with concentrations of 2 microg/ml or above. In conclusion, the broad spectrum of cytotoxicity of MKT 077 in the human tumor-cloning assay and the unique mechanism of action of MKT 077 encourage additional preclinical and clinical studies with this compound and other rhodacyanine dyes.

  4. Gentian violet exhibits activity against biofilms formed by oral Candida isolates obtained from HIV-infected patients.

    PubMed

    Traboulsi, Rana S; Mukherjee, Pranab K; Chandra, Jyotsna; Salata, Robert A; Jurevic, Richard; Ghannoum, Mahmoud A

    2011-06-01

    The effect of gentian violet against Candida albicans and non-Candida albicans biofilms formed on polymethylmethacrylate strips was evaluated using a dry weight assay and confocal laser scanning microscopy. The ability of gentian violet to inhibit Candida albicans germination was also assessed. Gentian violet activity against Candida biofilms was demonstrated by a reduction in dry weight, disruption of biofilm architecture, and reduced biofilm thickness. Additionally, gentian violet inhibited Candida germination in a concentration-dependent manner.

  5. Magnetohydrodynamic (MHD) analyses of various forms of activity and their propagation through helio spheric space

    NASA Technical Reports Server (NTRS)

    Wu, S. T.

    1987-01-01

    Theoretical and numerical modeling of solar activity and its effects on the solar atmosphere within the context of magnetohydrodynamics were examined. Specifically, the scientific objectives were concerned with the physical mechanisms for the flare energy build-up and subsequent release. In addition, transport of this energy to the corona and solar wind was also investigated. Well-posed, physically self-consistent, numerical simulation models that are based upon magnetohydrodynamics were sought. A systematic investigation of the basic processes that determine the macroscopic dynamic behavior of solar and heliospheric phenomena was conducted. A total of twenty-three articles were accepted and published in major journals. The major achievements are summarized.

  6. Influence of different forms of acidities on soil microbiological properties and enzyme activities at an acid mine drainage contaminated site.

    PubMed

    Sahoo, Prafulla Kumar; Bhattacharyya, Pradip; Tripathy, Subhasish; Equeenuddin, Sk Md; Panigrahi, M K

    2010-07-15

    Assessment of microbial parameters, viz. microbial biomass, fluorescence diacetate, microbial respiration, acid phosphatase, beta-glucosidase and urease with respect to acidity helps in evaluating the quality of soils. This study was conducted to investigate the effects of different forms of acidities on soil microbial parameters in an acid mine drainage contaminated site around coal deposits in Jainta Hills of India. Total potential and exchangeable acidity, extractable and exchangeable aluminium were significantly higher in contaminated soil compared to the baseline (p<0.01). Different forms of acidity were significantly and positively correlated with each other (p<0.05). Further, all microbial properties were positively and significantly correlated with organic carbon and clay (p<0.05). The ratios of microbial parameters with organic carbon were negatively correlated with different forms of acidity. Principal component analysis and cluster analyses showed that the microbial activities are not directly influenced by the total potential acidity and extractable aluminium. Though acid mine drainage affected soils had higher microbial biomass and activities due to higher organic matter content than those of the baseline soils, the ratios of microbial parameters/organic carbon indicated suppression of microbial growth and activities due to acidity stress.

  7. Activation of CD4+ T lymphocytes form interleukin 2-deficient mice by costimulatory B7 molecules.

    PubMed Central

    Razi-Wolf, Z; Höllander, G A; Reiser, H

    1996-01-01

    Interleukin 2 (IL-2)-deficient (IL-2-/-) mice develop hemolytic anemia and chronic inflammatory bowel disease. Importantly, the induction of disease in IL-2-deficient mice is critically dependent on CD4+ T cells. We have studied the requirements of T cells from IL-2-deficient mice for costimulation with B7 antigens. Stable B7-1 or B7-2 chinese hamster ovary (CHO) cell transfectants could synergize with anti-CD3 monoclonal antibody (mAb) to induce the proliferation of CD4+ T cells from IL-2-/- mutant mice. Further mechanistic studies established that B7-induced activation resulted in surface expression of the alpha chain of the IL-2 receptor. B7-induced proliferation occurred independently of IL-4 and was largely independent of the common gamma chain of the IL-2, IL-4, IL-7, IL-9, and IL-15 receptors. Finally, anti-B7-2 but not anti-B7-1 mAb was able to inhibit the activation of IL-2-/- T cells induced by anti-CD3 mAb in the presence of syngeneic antigen-presenting cells. The results of our experiments indicate that IL-2-/- CD4+ T cells remain responsive to B7 stimulation and raise the possibility that B7 antagonists have a role in the prevention/treatment of inflammatory bowel disease. Images Fig. 3 PMID:8610140

  8. Spectrofluorimetric determination of certain biologically active phenothiazines in commercial dosage forms and human plasma.

    PubMed

    Mohamed, Abdel-Maaboud I; Abdelmageed, Osama H; Salem, Hesham; Nagy, Dalia M; Omar, Mahmoud A

    2013-01-01

    A validated simple and sensitive spectrofluorimetric method was developed for the determination of chlorpromazine hydrochloride, promethazine hydrochloride, trifluperazine hydrochloride, thioridazine hydrochloride, perazine maleate and oxomemazine. The method was based on condensation of malonic acid/acetic anhydride (MAA) under the catalytic effect of the tertiary amine moiety of the studied phenothiazines to provide a deep yellow to brown colour with green fluorescence. Relative fluorescence intensity of the products was measured at λ exc 398 nm and λ em 432 nm. Different variables affecting the reaction were studied and optimized. The method was successfully applied for the determination of the studied drugs in commercial dosage forms. The lower detection limits allowed the application of this method for the determination of the compounds in plasma as an example of a biological fluid. In addition, the method was considered specific for the determination of tertiary amines in the presence of primary and secondary amines; as a result, it was deemed suitable for the determination of the cited drugs in the presence of their degradation products resulting from N-dealkylation or oxidation of the corresponding sulphoxides or sulphones.

  9. Amphipathic polymers: tools to fold integral membrane proteins to their active form.

    PubMed

    Pocanschi, Cosmin L; Dahmane, Tassadite; Gohon, Yann; Rappaport, Fabrice; Apell, Hans-Jürgen; Kleinschmidt, Jörg H; Popot, Jean-Luc

    2006-11-28

    Among the major obstacles to pharmacological and structural studies of integral membrane proteins (MPs) are their natural scarcity and the difficulty in overproducing them in their native form. MPs can be overexpressed in the non-native state as inclusion bodies, but inducing them to achieve their functional three-dimensional structure has proven to be a major challenge. We describe here the use of an amphipathic polymer, amphipol A8-35, as a novel environment that allows both beta-barrel and alpha-helical MPs to fold to their native state, in the absence of detergents or lipids. Amphipols, which are extremely mild surfactants, appear to favor the formation of native intramolecular protein-protein interactions over intermolecular or protein-surfactant ones. The feasibility of the approach is demonstrated using as models OmpA and FomA, two outer membrane proteins from the eubacteria Escherichia coli and Fusobacterium nucleatum, respectively, and bacteriorhodopsin, a light-driven proton pump from the plasma membrane of the archaebacterium Halobacterium salinarium.

  10. Systematic assessment of coordinated activity cliffs formed by kinase inhibitors and detailed characterization of activity cliff clusters and associated SAR information.

    PubMed

    Dimova, Dilyana; Stumpfe, Dagmar; Bajorath, Jürgen

    2015-01-27

    From currently available kinase inhibitors and their activity data, clusters of coordinated activity cliffs were systematically derived and subjected to cluster index and index map analysis. Type I-like inhibitors with well-defined IC50 measurements were found to provide a large knowledge base of activity cliff clusters for 266 targets from nine kinase groups. On the basis of index map analysis, these clusters were systematically organized according to structural similarity of inhibitors and activity cliff diversity and prioritized for structure-activity relationship (SAR) analysis. From prioritized clusters, interpretable SAR information can be extracted. It is also shown that activity cliff clusters formed by ATP site-directed inhibitors often represent local SAR environments of rather different complexity and interpretability. In addition, activity cliff clusters including promiscuous kinase inhibitors have been determined. Only a small subset of inhibitors was found to change activity cliff roles in different clusters. The activity cliff clusters described herein and their index map organization substantially enrich SAR information associated with kinase inhibitors in compound subsets of limited size. The cluster and index map information is made available upon request to provide opportunities for further SAR exploration. On the basis of our analysis and the data provided, activity cliff clusters and corresponding inhibitor series for kinase targets of interest can be readily selected.

  11. Structure of the unliganded form of the proprotein convertase furin suggests activation by a substrate-induced mechanism

    PubMed Central

    Dahms, Sven O.; Arciniega, Marcelino; Steinmetzer, Torsten; Huber, Robert; Than, Manuel E.

    2016-01-01

    Proprotein convertases (PCs) are highly specific proteases required for the proteolytic modification of many secreted proteins. An unbalanced activity of these enzymes is connected to pathologies like cancer, atherosclerosis, hypercholesterolaemia, and infectious diseases. Novel protein crystallographic structures of the prototypical PC family member furin in different functional states were determined to 1.8–2.0 Å. These, together with biochemical data and modeling by molecular dynamics calculations, suggest essential elements underlying its unusually high substrate specificity. Furin shows a complex activation mechanism and exists in at least four defined states: (i) the “off state,” incompatible with substrate binding as seen in the unliganded enzyme; (ii) the active “on state” seen in inhibitor-bound furin; and the respective (iii) calcium-free and (iv) calcium-bound forms. The transition from the off to the on state is triggered by ligand binding at subsites S1 to S4 and appears to underlie the preferential recognition of the four-residue sequence motif of furin. The molecular dynamics simulations of the four structural states reflect the experimental observations in general and provide approximations of the respective stabilities. Ligation by calcium at the PC-specific binding site II influences the active-site geometry and determines the rotamer state of the oxyanion hole-forming Asn295, and thus adds a second level of the activity modulation of furin. The described crystal forms and the observations of different defined functional states may foster the development of new tools and strategies for pharmacological intervention targeting furin. PMID:27647913

  12. Solubility Enhancement of a Poorly Water Soluble Drug by Forming Solid Dispersions using Mechanochemical Activation

    PubMed Central

    Rojas-Oviedo, I.; Retchkiman-Corona, B.; Quirino-Barreda, C. T.; Cárdenas, J.; Schabes-Retchkiman, P. S.

    2012-01-01

    Mechanochemical activation is a practical cogrinding operation used to obtain a solid dispersion of a poorly water soluble drug through changes in the solid state molecular aggregation of drug-carrier mixtures and the formation of noncovalent interactions (hydrogen bonds) between two crystalline solids such as a soluble carrier, lactose, and a poorly soluble drug, indomethacin, in order to improve its solubility and dissolution rate. Samples of indomethacin and a physical mixture with a weight ratio of 1:1 of indomethacin and lactose were ground using a high speed vibrating ball mill. Particle size was determined by electron microscopy, the reduction of crystallinity was determined by calorimetry and transmission electron microscopy, infrared spectroscopy was used to find evidence of any interactions between the drug and the carrier and the determination of apparent solubility allowed for the corroboration of changes in solubility. Before grinding, scanning electron microscopy showed the drug and lactose to have an average particle size of around 50 and 30 μm, respectively. After high speed grinding, indomethacin and the mixture had a reduced average particle size of around 5 and 2 μm, respectively, showing a morphological change. The ground mixture produced a solid dispersion that had a loss of crystallinity that reached 81% after 30 min of grinding while the drug solubility of indomethacin within the solid dispersion increased by 2.76 fold as compared to the pure drug. Drug activation due to hydrogen bonds between the carboxylic group of the drug and the hydroxyl group of lactose as well as the decrease in crystallinity of the solid dispersion and the reduction of the particle size led to a better water solubility of indomethacin. PMID:23798775

  13. Hsp90-binding immunophilin FKBP51 forms complexes with hTERT enhancing telomerase activity.

    PubMed

    Lagadari, Mariana; Zgajnar, Nadia R; Gallo, Luciana I; Galigniana, Mario D

    2016-08-01

    FK506-binding proteins are members of the immunophilin family of proteins. Those immunophilins associated to the 90-kDa-heat-shock protein, Hsp90, have been proposed as potential modulators of signalling cascade factors chaperoned by Hsp90. FKBP51 and FKBP52 are the best characterized Hsp90-bound immunophilins first described associated to steroid-receptors. The reverse transcriptase subunit of telomerase, hTERT, is also an Hsp90 client-protein and is highly expressed in cancer cells, where it is required to compensate the loss of telomeric DNA after each successive cell division. Because FKBP51 is also a highly expressed protein in cancer tissues, we analyzed its potential association with hTERT·Hsp90 complexes and its possible biological role. In this study it is demonstrated that both immunophilins, FKBP51 and FKBP52, co-immunoprecipitate with hTERT. The Hsp90 inhibitor radicicol disrupts the heterocomplex and favors the partial cytoplasmic relocalization of hTERT in similar manner as the overexpression of the TPR-domain peptide of the immunophilin. While confocal microscopy images show that FKBP51 is primarily localized in mitochondria and hTERT is totally nuclear, upon the onset of oxidative stress, FKBP51 (but not FKBP52) becomes mostly nuclear colocalizing with hTERT, and longer exposure times to peroxide favors hTERT export to mitochondria. Importantly, telomerase activity of hTERT is significantly enhanced by FKBP51. These observations support the emerging role assigned to FKBP51 as antiapoptotic factor in cancer development and progression, and describe for the first time the potential role of this immunophilin favoring the clonal expansion by enhancing telomerase activity.

  14. Activity of a Carboxyl-Terminal Truncated Form of Catechol 2,3-Dioxygenase from Planococcus sp. S5

    PubMed Central

    2014-01-01

    Catechol 2,3-dioxygenases (C23Os, E.C.1.13.12.2) are two domain enzymes that catalyze degradation of monoaromatic hydrocarbons. The catalytically active C-domain of all known C23Os comprises ferrous ion ligands as well as residues forming active site pocket. The aim of this work was to examine and discuss the effect of nonsense mutation at position 289 on the activity of catechol 2,3-dioxygenase from Planococcus strain. Although the mutant C23O showed the same optimal temperature for activity as the wild-type protein (35°C), it exhibited activity slightly more tolerant to alkaline pH. Mutant enzyme exhibited also higher affinity to catechol as a substrate. Its Km (66.17 µM) was approximately 30% lower than that of wild-type enzyme. Interestingly, removal of the C-terminal residues resulted in 1.5- to 1.8-fold (P < 0.05) increase in the activity of C23OB61 against 4-methylcatechol and 4-chlorocatechol, respectively, while towards catechol the activity of the protein dropped to about 80% of that of the wild-type enzyme. The results obtained may facilitate the engineering of the C23O for application in the bioremediation of polluted areas. PMID:24693238

  15. Human liver mitochondrial aldehyde dehydrogenase: three-dimensional structure and the restoration of solubility and activity of chimeric forms.

    PubMed Central

    Ni, L.; Zhou, J.; Hurley, T. D.; Weiner, H.

    1999-01-01

    Human liver cytosolic and mitochondrial isozymes of aldehyde dehydrogenase share 70% sequence identity. However, the first 21 residues are not conserved between the human isozymes (15% identity). The three-dimensional structures of the beef mitochondrial and sheep cytosolic forms have virtually identical three-dimensional structures. Here, we solved the structure of the human mitochondrial enzyme and found it to be identical to the beef enzyme. The first 21 residues are found on the surface of the enzyme and make no contact with other subunits in the tetramer. A pair of chimeric enzymes between the human isozymes was made. Each chimera had the first 21 residues from one isozyme and the remaining 479 from the other. When the first 21 residues were from the mitochondrial isozyme, an enzyme with cytosolic-like properties was produced. The other was expressed but was insoluble. It was possible to restore solubility and activity to the chimera that had the first 21 cytosolic residues fused to the mitochondrial ones by making point mutations to residues at the N-terminal end. When residue 19 was changed from tyrosine to a cysteine, the residue found in the mitochondrial form, an active enzyme could be made though the Km for NAD+ was 35 times higher than the native mitochondrial isozyme and the specific activity was reduced by 75%. This residue interacts with residue 203, a nonconserved, nonactive site residue. A mutation of residue 18, which also interacts with 203, restored solubility, but not activity. Mutation to residue 15, which interacts with 104, also restored solubility but not activity. It appears that to have a soluble or active enzyme a favorable interaction must occur between a residue in a surface loop and a residue elsewhere in the molecule even though neither make contact with the active site region of the enzyme. PMID:10631996

  16. Waste Form Release Data Package for the 2001 Immobilized Low-Activity Waste Performance Assessment

    SciTech Connect

    McGrail, B. Peter; Icenhower, Jonathan P.; Martin, Paul F.; Schaef, Herbert T.; O'Hara, Matthew J.; Rodriguez, Eugenio; Steele, Jackie L.

    2001-02-01

    This data package documents the experimentally derived input data on the representative waste glasses LAWABP1 and HLP-31 that will be used for simulations of the immobilized lowactivity waste disposal system with the Subsurface Transport Over Reactive Multiphases (STORM) code. The STORM code will be used to provide the near-field radionuclide release source term for a performance assessment to be issued in March of 2001. Documented in this data package are data related to 1) kinetic rate law parameters for glass dissolution, 2) alkali-H ion exchange rate, 3) chemical reaction network of secondary phases that form in accelerated weathering tests, and 4) thermodynamic equilibrium constants assigned to these secondary phases. The kinetic rate law and Na+-H+ ion exchange rate were determined from single-pass flow-through experiments. Pressurized unsaturated flow and vapor hydration experiments were used for accelerated weathering or aging of the glasses. The majority of the thermodynamic data were extracted from the thermodynamic database package shipped with the geochemical code EQ3/6. However, several secondary reaction products identified from laboratory tests with prototypical LAW glasses were not included in this database, nor are the thermodynamic data available in the open literature. One of these phases, herschelite, was determined to have a potentially significant impact on the release calculations and so a solubility product was estimated using a polymer structure model developed for zeolites. Although this data package is relatively complete, final selection of ILAW glass compositions has not been done by the waste treatment plant contractor. Consequently, revisions to this data package to address new ILAW glass formulations are to be regularly expected.

  17. Channel-forming activity of syringomycin E in two mercury-supported biomimetic membranes.

    PubMed

    Becucci, Lucia; Tramonti, Vania; Fiore, Alberto; Fogliano, Vincenzo; Scaloni, Andrea; Guidelli, Rolando

    2015-04-01

    The lipodepsipeptide syringomycin E (SR-E) interacts with two mercury-supported biomimetic membranes, which consist of a self-assembled phospholipid monolayer (SAM) and of a tethered bilayer lipid membrane (tBLM) separated from the mercury surface by a hydrophilic tetraethyleneoxy (TEO) spacer that acts as an ionic reservoir. SR-E interacts more rapidly and effectively with a SAM of dioleoylphosphatidylserine (DOPS) than with one of dioleoylphosphatidylcholine (DOPC). The proximal lipid monolayer of the tBLM has no polar head region, being linked to the TEO spacer via an ether bond, while the distal monolayer consists of either a DOPC or a DOPS leaflet. The ion flow into or out of the spacer through the lipid bilayer moiety of the tBLM was monitored by potential step chronocoulometry and cyclic voltammetry. With the distal monolayer bathed by aqueous 0.1M KCl and 0.8μM SR-E, an ion flow in two stages was monitored with DOPC at pH3 and 5.4 and with DOPS at pH3, while a single stage was observed with DOPS at pH5.4. This behavior was compared with that already described at conventional bilayer lipid membranes. The sigmoidal shape of the chronocoulometric charge transients points to an aggregation of SR-E monomers forming an ion channel via a mechanism of nucleation and growth. The ion flow is mainly determined by potassium ions, and is inhibited by calcium ions. The contribution to the transmembrane potential from the distal leaflet depends more on the nature of the lipid than that of the ion channel.

  18. Tooth development in Ambystoma mexicanum: phosphatase activities, calcium accumulation and cell proliferation in the tooth-forming tissues.

    PubMed

    Wistuba, Joachim; Ehmcke, Jens; Clemen, Günter

    2003-06-01

    Prerequisites of tooth formation, cell proliferation in the tooth-forming tissues, calcium accumulation and the enzymatic activities of alkaline (ALP) and acid phosphatases (ACP) were investigated by immunohistochemical and histochemical methods in various developmental stages of the Mexican Axolotl, Ambystoma mexicanum. During the growth of replacement teeth, the tooth-forming tissues continually recruit cells from the surrounding regions. The basal layer of the oral epithelium, the dental lamina and sometimes even the outer enamel epithelium provide cells for the differentiated inner enamel epithelium, in which the active ameloblasts are localized. The differentiating odontoblasts are derived from proliferating cells situated basally to the replacement teeth in the mesenchymal tissue. When differentiation has started and the cells have become functional, proliferative activity can no longer be observed. Calcium is accumulated close to the site of mineralization in the inner enamel epithelium and in the odontoblasts as it is in mammals, elasmobranchii and teleostei. The activities of ACP and ALP related to the mineralization of the replacement teeth are separated spatially and not sequentially as they are in mammals. However, the results indicate a similar function of these enzymatic components in relation to tooth formation and maturation of mineral deposition. Most of the substantial processes related to tooth formation reported from other vertebrates occur in a manner similar to that in Ambystoma mexicanum, but there also seem to be basic mechanisms present that are realised in a unique way in this urodele.

  19. PLTP secreted by HepG2 cells resembles the high-activity PLTP form in human plasma.

    PubMed

    Siggins, Sarah; Jauhiainen, Matti; Olkkonen, Vesa M; Tenhunen, Jukka; Ehnholm, Christian

    2003-09-01

    Plasma phospholipid transfer protein (PLTP) is an important regulator of plasma HDL levels and HDL particle distribution. PLTP is present in plasma in two forms, one with high and the other with low phospholipid transfer activity. We have used the human hepatoma cell line, HepG2, as a model to study PLTP secreted from hepatic cells. PLTP activity was secreted by the cells into serum-free culture medium as a function of time. However, modification of a previously established ELISA assay to include a denaturing sample pretreatment with the anionic detergent sodium dodecyl sulphate was required for the detection of the secreted PLTP protein. The HepG2 PLTP could be enriched by Heparin-Sepharose affinity chromatography and eluted in size-exclusion chromatography at a position corresponding to the size of 160 kDa. PLTP coeluted with apolipoprotein E (apoE) but not with apoB-100 or apoA-I. A portion of PLTP was retained by an anti-apoE immunoaffinity column together with apoE, suggesting an interaction between these two proteins. Furthermore, antibodies against apoE but not those against apoB-100 or apoA-I were capable of inhibiting PLTP activity. These results show that the HepG2-derived PLTP resembles in several aspects the high-activity form of PLTP found in human plasma.

  20. Fluidized bed steam reformed mineral waste form performance testing to support Hanford Supplemental Low Activity Waste Immobilization Technology Selection

    SciTech Connect

    Jantzen, C. M.; Pierce, E. M.; Bannochie, C. J.; Burket, P. R.; Cozzi, A. D.; Crawford, C. L.; Daniel, W. E.; Fox, K. M.; Herman, C. C.; Miller, D. H.; Missimer, D. M.; Nash, C. A.; Williams, M. F.; Brown, C. F.; Qafoku, N. P.; Neeway, J. J.; Valenta, M. M.; Gill, G. A.; Swanberg, D. J.; Robbins, R. A.; Thompson, L. E.

    2015-10-01

    This report describes the benchscale testing with simulant and radioactive Hanford Tank Blends, mineral product characterization and testing, and monolith testing and characterization. These projects were funded by DOE EM-31 Technology Development & Deployment (TDD) Program Technical Task Plan WP-5.2.1-2010-001 and are entitled “Fluidized Bed Steam Reformer Low-Level Waste Form Qualification”, Inter-Entity Work Order (IEWO) M0SRV00054 with Washington River Protection Solutions (WRPS) entitled “Fluidized Bed Steam Reforming Treatability Studies Using Savannah River Site (SRS) Low Activity Waste and Hanford Low Activity Waste Tank Samples”, and IEWO M0SRV00080, “Fluidized Bed Steam Reforming Waste Form Qualification Testing Using SRS Low Activity Waste and Hanford Low Activity Waste Tank Samples”. This was a multi-organizational program that included Savannah River National Laboratory (SRNL), THOR® Treatment Technologies (TTT), Pacific Northwest National Laboratory (PNNL), Oak Ridge National Laboratory (ORNL), Office of River Protection (ORP), and Washington River Protection Solutions (WRPS). The SRNL testing of the non-radioactive pilot-scale Fluidized Bed Steam Reformer (FBSR) products made by TTT, subsequent SRNL monolith formulation and testing and studies of these products, and SRNL Waste Treatment Plant Secondary Waste (WTP-SW) radioactive campaign were funded by DOE Advanced Remediation Technologies (ART) Phase 2 Project in connection with a Work-For-Others (WFO) between SRNL and TTT.

  1. The active form of the norovirus RNA-dependent RNA polymerase is a homodimer with cooperative activity.

    PubMed

    Högbom, Martin; Jäger, Katrin; Robel, Ivonne; Unge, Torsten; Rohayem, Jacques

    2009-02-01

    Norovirus (NV) is a leading cause of gastroenteritis worldwide and a major public health concern. So far, the replication strategy of NV remains poorly understood, mainly because of the lack of a cell system to cultivate the virus. In this study, the function and the structure of a key viral enzyme of replication, the RNA-dependent RNA polymerase (RdRp, NS7), was examined. The overall structure of the NV NS7 RdRp was determined by X-ray crystallography to a 2.3 A (0.23 nm) resolution (PDB ID 2B43), displaying a right-hand fold typical of the template-dependent polynucleotide polymerases. Biochemical analysis evidenced that NV NS7 RdRp is active as a homodimer, with an apparent K(d) of 0.649 microM and a positive cooperativity (Hill coefficient n(H)=1.86). Crystals of the NV NS7 homodimer displayed lattices containing dimeric arrangements with high shape complementarity statistics. This experimental data on the structure and function of the NV RdRp may set the cornerstone for the development of polymerase inhibitors to control the infection with NV, a medically relevant pathogen.

  2. Fate and Transport of Methane Formed in the Active Layer of Alaskan Permafrost

    NASA Astrophysics Data System (ADS)

    Conrad, M. E.; Curtis, J. B.; Smith, L. J.; Bill, M.; Torn, M. S.

    2015-12-01

    Over the past 2 years a series of tracer tests designed to estimate rates of methane formation via acetoclastic methanogenesis in the active layer of permafrost soils were conducted at the Barrow Environmental Observatory (BEO) in northernmost Alaska. The tracer tests consisted of extracting 0.5 to 1.0 liters of soil water in gas-tight bags from different features of polygons at the BEO, followed by addition of a tracer cocktail including acetate with a 13C-labeled methyl group and D2O (as a conservative tracer) into the soil water and injection of the mixture back into the original extraction site. Samples were then taken at depths of 30 cm (just above the bottom of the active layer), 20 cm, 10 cm and surface flux to determine the fate of the 13C-labeled acetate. During 2014 (2015 results are pending) water, soil gas, and flux gas were sampled for 60 days following injection of the tracer solution. Those samples were analyzed for concentrations and isotopic compositions of CH4, DIC/CO2 and water. At one site (the trough of a low-centered polygon) the 13C acetate was completely converted to 13CH4 within the first 2 days. The signal persisted for throughout the entire monitoring period at the injection depth with little evidence of transport or oxidation in any of the other sampling depths. In the saturated center of the same polygon, the acetate was also rapidly converted to 13CH4, but water turnover caused the signal to rapidly dissipate. High δ13C CO2 in flux samples from the polygon center indicate oxidation of the 13CH4 in near-surface waters. Conversely, CH4 production in the center of an unsaturated, flat-centered polygon was relatively small 13CH4 and dissipated rapidly without any evidence of either 13CH4 transport to shallower levels or oxidation. At another site in the edge of that polygon no 13CH4 was produced, but significant 13CO2/DIC was observed indicating direct aerobic oxidation of the acetate was occurring at this site. These results suggest that

  3. Dependence of photosynthesis and energy dissipation activity upon growth form and light environment during the winter.

    PubMed

    Adams, W W; Demmig-Adams, B; Rosenstiel, T N; Ebbert, V

    2001-01-01

    Two very distinctive responses of photosynthesis to winter conditions have been identified. Mesophytic species that continue to exhibit growth during the winter typically exhibit higher maximal rates of photosynthesis during the winter or when grown at lower temperatures compared to individuals examined during the summer or when grown at warmer temperatures. In contrast, sclerophytic evergreen species growing in sun-exposed sites typically exhibit lower maximal rates of photosynthesis in the winter compared to the summer. On the other hand, shaded individuals of those same sclerophytic evergreen species exhibit similar or higher maximal rates of photosynthesis in the winter compared to the summer. Employment of the xanthophyll cycle in photoprotective energy dissipation exhibits similar characteristics in the two groups of plants (mesophytes and shade leaves of sclerophytic evergreens) that exhibit upregulation of photosynthesis during the winter. In both, zeaxanthin + antheraxanthin (Z + A) are retained and PS II remains primed for energy dissipation only on nights with subfreezing temperatures, and this becomes rapidly reversed upon exposure to increased temperatures. In contrast, Z + A are retained and PS II remains primed for energy dissipation over prolonged periods during the winter in sun leaves of sclerophytic evergreen species, and requires days of warming to become fully reversed. The rapid disengagement of this energy dissipation process in the mesophytes and shade sclerophytes apparently permits a rapid return to efficient photosynthesis and increased activity on warmer days during the winter. This may be associated with a decreasing opportunity for photosynthesis in source leaves relative to the demand for photosynthesis in the plant's sinks. In contrast, the sun-exposed sclerophytes - with a relatively high source to sink ratio - maintain PS II in a state primed for high levels of energy dissipation activity throughout much of the winter. Independent

  4. Structure, Aggregation, and Activity of a Covalent Insulin Dimer Formed During Storage of Neutral Formulation of Human Insulin.

    PubMed

    Hjorth, Christian Fogt; Norrman, Mathias; Wahlund, Per-Olof; Benie, Andrew J; Petersen, Bent O; Jessen, Christian M; Pedersen, Thomas Å; Vestergaard, Kirsten; Steensgaard, Dorte B; Pedersen, Jan Skov; Naver, Helle; Hubálek, František; Poulsen, Christian; Otzen, Daniel

    2016-04-01

    A specific covalently linked dimeric species of insulin high molecular weight products (HMWPs), formed during prolonged incubation of a neutral pharmaceutical formulation of human insulin, were characterized in terms of tertiary structure, self-association, biological activity, and fibrillation properties. The dimer was formed by a covalent link between A21Asn and B29Lys. It was analyzed using static and dynamic light scattering and small-angle X-ray scattering to evaluate its self-association behavior. The tertiary structure was obtained using nuclear magnetic resonance and X-ray crystallography. The biological activity of HMWP was determined using 2 in vitro assays, and its influence on fibrillation was investigated using Thioflavin T assays. The dimer's tertiary structure was nearly identical to that of the noncovalent insulin dimer, and it was able to form hexamers in the presence of zinc. The dimer exhibited reduced propensity for self-association in the absence of zinc but significantly postponed the onset of fibrillation in insulin formulations. Consistent with its dimeric state, the tested species of HMWP showed little to no biological activity in the used assays. This study is the first detailed characterization of a specific type of human insulin HMWP formed during storage of a marketed pharmaceutical formulation. These results indicate that this specific type of HMWP is unlikely to antagonize the physical stability of the formulation, as HMWP retained a tertiary structure similar to the noncovalent dimer and participated in hexamer assembly in the presence of zinc. In addition, increasing amounts of HMWP reduce the rate of insulin fibrillation.

  5. Constitutively active form of natriuretic peptide receptor 2 ameliorates experimental pulmonary arterial hypertension

    PubMed Central

    Nawa, Nobutoshi; Ishida, Hidekazu; Katsuragi, Shinichi; Baden, Hiroki; Takahashi, Kunihiko; Higeno, Ryota; Torigoe, Fumiko; Mihara, Seiko; Narita, Jun; Miura, Kohji; Nakamura, Kazufumi; Kogaki, Shigetoyo; Ozono, Keiichi

    2016-01-01

    We recently found a constitutively active mutant of natriuretic peptide receptor 2 (caNPR2; V883M), which synthesizes larger amounts of cyclic guanosine monophosphate (cGMP) intracellularly without any ligand stimulation than existing drugs. The aim of this study was to investigate the therapeutic effects of gene transduction using caNPR2 for pulmonary arterial hypertension (PAH). In vitro gene transduction into human pulmonary arterial smooth muscle cells using Sendai virus (SeV) vectors carrying caNPR2 induced 10,000-fold increases in the synthesis of cGMP without ligand stimulation, and the proliferation of caNPR2-expressing cells was significantly attenuated. The PAH model rats generated by hypoxia and the administration of SU5416 were then treated with SeV vectors through a direct injection into the left pulmonary artery. Right ventricular systolic pressure was significantly decreased 2 weeks after the treatment, while systemic blood pressure remained unchanged. Histological analyses revealed that the medial wall thickness and occlusion rate of pulmonary arterioles were significantly improved in caNPR2-treated lungs. Neither the systemic integration of virus vectors nor side effects were observed. The massive stimulation of cGMP synthesis by gene therapy with caNPR2 was safe and effective in a PAH rat model and, thus, has potential as a novel therapy for patients with severe progressive PAH. PMID:27419193

  6. Constitutively active form of natriuretic peptide receptor 2 ameliorates experimental pulmonary arterial hypertension.

    PubMed

    Nawa, Nobutoshi; Ishida, Hidekazu; Katsuragi, Shinichi; Baden, Hiroki; Takahashi, Kunihiko; Higeno, Ryota; Torigoe, Fumiko; Mihara, Seiko; Narita, Jun; Miura, Kohji; Nakamura, Kazufumi; Kogaki, Shigetoyo; Ozono, Keiichi

    2016-01-01

    We recently found a constitutively active mutant of natriuretic peptide receptor 2 (caNPR2; V883M), which synthesizes larger amounts of cyclic guanosine monophosphate (cGMP) intracellularly without any ligand stimulation than existing drugs. The aim of this study was to investigate the therapeutic effects of gene transduction using caNPR2 for pulmonary arterial hypertension (PAH). In vitro gene transduction into human pulmonary arterial smooth muscle cells using Sendai virus (SeV) vectors carrying caNPR2 induced 10,000-fold increases in the synthesis of cGMP without ligand stimulation, and the proliferation of caNPR2-expressing cells was significantly attenuated. The PAH model rats generated by hypoxia and the administration of SU5416 were then treated with SeV vectors through a direct injection into the left pulmonary artery. Right ventricular systolic pressure was significantly decreased 2 weeks after the treatment, while systemic blood pressure remained unchanged. Histological analyses revealed that the medial wall thickness and occlusion rate of pulmonary arterioles were significantly improved in caNPR2-treated lungs. Neither the systemic integration of virus vectors nor side effects were observed. The massive stimulation of cGMP synthesis by gene therapy with caNPR2 was safe and effective in a PAH rat model and, thus, has potential as a novel therapy for patients with severe progressive PAH.

  7. Use of silicon carbide sludge to form porous alkali-activated materials for insulating application

    NASA Astrophysics Data System (ADS)

    Prud'homme, E.; Joussein, E.; Rossignol, S.

    2015-07-01

    One of the objectives in the field of alkali-activated materials is the development of materials having greater thermal performances than conventional construction materials such as aerated concrete. The aim of this paper is to present the possibility to obtain controlled porosity and controlled thermal properties with geopolymer materials including a waste like silicon carbide sludge. The porosity is created by the reaction of free silicon contains in silicon carbide sludge leading to the formation of hydrogen. Two possible ways are investigated to control the porosity: modification of mixture formulation and additives introduction. The first way is the most promising and allowed the formation of materials presenting the same density but various porosities, which shows that the material is adaptable to the application. The insulation properties are logically linked to the porosity and density of materials. A lower value of thermal conductivity of 0.075 W.m-1.K-1 can be reached for a material with a low density of 0.27 g.cm-3. These characteristics are really good for a mineral-based material which always displays non-negligible resistance to manipulation.

  8. Artefactual nanoparticle activation of the inflammasome platform: in vitro evidence with a nano-formed calcium phosphate

    PubMed Central

    Pele, Laetitia; Haas, Carolin T; Hewitt, Rachel; Faria, Nuno; Brown, Andy; Powell, Jonathan

    2015-01-01

    Aim To determine whether in vitro experimental conditions dictate cellular activation of the inflammasome by apatitic calcium phosphate nanoparticles. Material & methods The responses of blood-derived primary human cells to in situ-formed apatite were investigated under different experimental conditions to assess the effect of aseptic culture, cell rest and duration of particle exposure. Cell death and particle uptake were assessed, while IL-1β and caspase 1 responses, with and without lipopolysaccharide prestimulation, were evaluated as markers of inflammasome activation. Results Under carefully addressed experimental conditions, apatitic nanoparticles did not induce cell death or engage the inflammasome platform, although both could be triggered through artefacts of experimentation. Conclusion In vitro studies often predict that engineered nanoparticles, such as synthetic apatite, are candidates for inflammasome activation and, hence, are toxic. However, the experimental setting must be very carefully considered as it may promote false-positive outcomes. PMID:24991724

  9. Spontaneous activity in electromyography may differentiate certain benign lower motor neuron disease forms from amyotrophic lateral sclerosis.

    PubMed

    Jokela, Manu E; Jääskeläinen, Satu K; Sandell, Satu; Palmio, Johanna; Penttilä, Sini; Saukkonen, Annamaija; Soikkeli, Raija; Udd, Bjarne

    2015-08-15

    There is limited data on electromyography (EMG) findings in other motor neuron disorders than amyotrophic lateral sclerosis (ALS). We assessed whether the distribution of active denervation detected by EMG, i.e. fibrillations and fasciculations, differs between ALS and slowly progressive motor neuron disorders. We compared the initial EMG findings of 43 clinically confirmed, consecutive ALS patients with those of 41 genetically confirmed Late-onset Spinal Motor Neuronopathy and 14 Spinal and Bulbar Muscular Atrophy patients. Spontaneous activity was more frequently detected in the first dorsal interosseus and deltoid muscles of ALS patients than in patients with the slowly progressive motor neuron diseases. The most important observation was that absent fibrillations in the first dorsal interosseus muscle identified the benign forms with sensitivities of 66%-77% and a specificity of 93%. The distribution of active denervation may help to separate ALS from mimicking disorders at an early stage.

  10. [Kinetic modeling of energy metabolism and generation of active forms of oxygen in hepatocyte mitochondria].

    PubMed

    Demin, O V; Gorianin, I I; Kholodenko, B N; Westerhoff, H V

    2001-01-01

    Direct nonenzymatic oxidation of semiquinone by oxygen is one of the main sources of superoxide radicals (O2.-) in mitochondria. By using all the known data on hepatocyte mitochondria, we have revealed the correlation between the rate of superoxide generation by the bc1 complex and the transmembrane potential (delta psi). If the main electrogenic stage of the Q cycle is suggested to be the electron transfer between the cytochrome b hemes, then the rate of superoxide generation sharply increases when delta psi grows from 150 mV to 180 mV. However, this interrelation is ambiguous. Indeed, the increase of the generation rate with the growth of the potential can occur faster when succinate dehydrogenase is inhibited by malonate than when external ADP is exhausted. When the potential is changed by adding phosphate or potassium (K+), the rate of O2.- production remains constant, although the comparison of the rate values at the same delta psi reveals the effect of phosphate or potassium. It turned out that the rate of O2.- generation is a function of delta mu H rather than any of its components. Phosphate and K+ have practically no influence on delta mu H, since the change in delta psi is compensated by delta pH. The rate of superoxide generation by the bc1 complex is a multiple function of the electron-transfer activity of enzymes, the processes determining the membrane potential (e.g., loading), and of the oxygen concentration. The kinetic model proposed in this work may serve a tool to understand how the superoxide production is regulated.

  11. The biologically active form of the sea urchin egg receptor for sperm is a disulfide-bonded homo-multimer

    PubMed Central

    1994-01-01

    Since many cell surface receptors exist in their active form as oligomeric complexes, we have investigated the subunit composition of the biologically active sperm receptor in egg plasma membranes from Strongylocentrotus purpuratus. Electrophoretic analysis of the receptor without prior reduction of disulfide bonds revealed that the surface receptor exists in the form of a disulfide-bonded multimer, estimated to be a tetramer. These findings are in excellent agreement with the fact that the NH2-terminus of the extracellular domain of the sperm receptor is rich in cysteine residues. Studies with cross-linking agents of various length and hydrophobicity suggest that no other major protein is tightly associated with the receptor. Given the multimeric structure of the receptor, we investigated the effect of disulfide bond reduction on its biological activity. Because in quantitative bioassays fertilization was found to be inhibited by treatment of eggs with 5 mM dithiothreitol, we undertook more direct studies of the effect of reduction on properties of the receptor. First, we studied the effect of addition of isolated, pure receptor on fertilization. Whereas the non-reduced, native receptor complex inhibited fertilization in a dose- dependent manner, the reduced and alkylated receptor was inactive. Second, we tested the ability of the isolated receptor to mediate binding of acrosome-reacted sperm to polystyrene beads. Whereas beads coated with native receptor bound sperm, those containing reduced and alkylated receptor did not. Thus, these results demonstrate that the biologically active form of the sea urchin sperm receptor consists only of 350 kD subunits and that these must be linked as a multimer via disulfide bonds to produce a complex that is functional in sperm recognition and binding. PMID:8188748

  12. Demonstration of interleukin 1 activity in apparently homogeneous specimens of the pI 5 form of rabbit endogenous pyrogen.

    PubMed

    Hanson, D F; Murphy, P A

    1984-08-01

    Rabbit mononuclear cells from oil-induced peritoneal exudates were purified by centrifugation on Percoll gradients, suspended in tissue culture medium, and stimulated with opsonized Staphylococcus epidermidis. The supernatants from these macrophages caused fever when injected intravenously into rabbits (endogenous pyrogen [EP] activity). The EP activity was contained in two protein fractions, with pIs of 7.3 and ca. 5.0. The same fractions caused mouse thymocytes to incorporate tritiated thymidine when incubated in vitro with small quantities of phytohemagglutinin (interleukin 1 [IL-1] activity). The pI 5.0 form of EP was purified to apparent homogeneity by sequential use of ammonium sulfate precipitation, gel filtration, ion-exchange chromatography, hydrophobic chromatography, and high-resolution isoelectric focusing. EP and IL-1 activities were not separable by any of these procedures. Active fractions from isoelectric focusing were analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Only one band was visible as judged by a silver staining method, and IL-1 activity could be recovered by renaturing eluates from the same region of sodium dodecyl sulfate gels run in parallel. An estimate of specific activity was made by comparing the intensity of stained bands of EP with the intensity of bands containing known quantities of lysozyme or RNase. By this criterion, the specific activity of purified pI 5 EP was between 17,000 and 58,000 degrees C U/mg of protein, and the specific activity in terms of IL-1 was between 59 million and 360 million U per mg of protein. These observations suggest that both EP and IL-1 activities can be expressed by a single molecular species. The implications of this coincidence are discussed. It was also shown that highly purified pI 5 EP obtained from macrophages stimulated in the presence of 14C-labeled amino acids contained significant 14C radioactivity. This suggests that the pI 5.0 EP, like the pI 7

  13. Bimolecular recombination reactions: K-adiabatic and K-active forms of the bimolecular master equations and analytic solutions.

    PubMed

    Ghaderi, Nima

    2016-03-28

    Expressions for a K-adiabatic master equation for a bimolecular recombination rate constant krec are derived for a bimolecular reaction forming a complex with a single well or complexes with multiple well, where K is the component of the total angular momentum along the axis of least moment of inertia of the recombination product. The K-active master equation is also considered. The exact analytic solutions, i.e., the K-adiabatic and K-active steady-state population distribution function of reactive complexes, g(EJK) and g(EJ), respectively, are derived for the K-adiabatic and K-active master equation cases using properties of inhomogeneous integral equations (Fredholm type). The solutions accommodate arbitrary intermolecular energy transfer models, e.g., the single exponential, double exponential, Gaussian, step-ladder, and near-singularity models. At the high pressure limit, the krec for both the K-adiabatic and K-active master equations reduce, respectively, to the K-adiabatic and K-active bimolecular Rice-Ramsperger-Kassel-Marcus theory (high pressure limit expressions). Ozone and its formation from O + O2 are known to exhibit an adiabatic K. The ratio of the K-adiabatic to the K-active recombination rate constants for ozone formation at the high pressure limit is calculated to be ∼0.9 at 300 K. Results on the temperature and pressure dependence of the recombination rate constants and populations of O3 will be presented elsewhere.

  14. Identification of two forms of Q{beta} replicase with different thermal stabilities but identical RNA replication activity.

    PubMed

    Ichihashi, Norikazu; Matsuura, Tomoaki; Hosoda, Kazufumi; Yomo, Tetsuya

    2010-11-26

    The enzyme Qβ replicase is an RNA-dependent RNA polymerase, which plays a central role in infection by the simple single-stranded RNA virus bacteriophage Qβ. This enzyme has been used in a number of applications because of its unique activity in amplifying RNA from an RNA template. Determination of the thermal stability of Qβ replicase is important to gain an understanding of its function and potential applications, but data reported to date have been contradictory. Here, we provide evidence that these previous inconsistencies were due to the heterogeneous forms of the replicase with different stabilities. We purified two forms of replicase expressed in Escherichia coli, which differed in their thermal stability but showed identical RNA replication activity. Furthermore, we found that the replicase undergoes conversion between these forms due to oxidation, and the Cys-533 residue in the catalytic β subunit and Cys-82 residue in the EF-Tu subunit of the replicase are essential prerequisites for this conversion to occur. These results strongly suggest that the thermal stable replicase contains the intersubunit disulfide bond between these cysteines. The established strategies for isolating and purifying a thermally stable replicase should increase the usefulness of Qβ replicase in various applications, and the data regarding thermal stability obtained in this study may yield insight into the precise mechanism of infection by bacteriophage Qβ.

  15. Study of the Protein Complex, Pore Diameter, and Pore-forming Activity of the Borrelia burgdorferi P13 Porin*

    PubMed Central

    Bárcena-Uribarri, Iván; Thein, Marcus; Barbot, Mariam; Sans-Serramitjana, Eulalia; Bonde, Mari; Mentele, Reinhard; Lottspeich, Friedrich; Bergström, Sven; Benz, Roland

    2014-01-01

    P13 is one of the major outer membrane proteins of Borrelia burgdorferi. Previous studies described P13 as a porin. In the present study some structure and function aspects of P13 were studied. P13 showed according to lipid bilayer studies a channel-forming activity of 0.6 nanosiemens in 1 m KCl. Single channel and selectivity measurements demonstrated that P13 had no preference for either cations or anions and showed no voltage-gating up to ±100 mV. Blue native polyacrylamide gel electrophoresis was used to isolate and characterize the P13 protein complex in its native state. The complex had a high molecular mass of about 300 kDa and was only composed of P13 monomers. The channel size was investigated using non-electrolytes revealing an apparent diameter of about 1.4 nm with a 400-Da molecular mass cut-off. Multichannel titrations with different substrates reinforced the idea that P13 forms a general diffusion channel. The identity of P13 within the complex was confirmed by second dimension SDS-PAGE, Western blotting, mass spectrometry, and the use of a p13 deletion mutant strain. The results suggested that P13 is the protein responsible for the 0.6-nanosiemens pore-forming activity in the outer membrane of B. burgdorferi. PMID:24825899

  16. Antibacterial Activity of Euphorbia hebecarpa Alcoholic Extracts Against Six Human Pathogenic Bacteria in Planktonic and Biofilm Forms

    PubMed Central

    Mohsenipour, Zeinab; Hassanshahian, Mehdi

    2016-01-01

    Background Biofilm formation is a primary cause of considerable bacterial destruction. Objectives In an effort to combat these industrial and medical bacterial biofilm problems, our study aims to determine the antimicrobial effect of Euphorbia hebecarpa. Materials and Methods The inhibition efficiency of alcoholic extracts on the planktonic form of six pathogenic bacteria was evaluated using a disk diffusion technique. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) values were determined by means of a macrobroth dilution method. The effects of the extracts on biofilms were calculated using a microtiter plate method. Results The results of the disk diffusion assay (MBC and MIC) confirmed that E. hebecarpa ethanolic extracts were more efficient than methanolic extracts in the inhibition of planktonic forms of bacteria. Also, the inhibitory effect of the extracts in a broth medium was greater than in a solid medium. Extracts of E. hebecarpa were found to inhibit biofilm formation better than demolish of biofilm and preventing metabolic activity of bacteria in biofilm structures. The greatest inhibitory effects of E. hebecarpa extracts were observed for the biofilm formation of B. cereus (92.81%). In addition, the greatest demolition was observed for the S. aureus biofilm (74.49%), and the metabolic activity decrement of this bacteria was highest (78.21%) of all the tested bacteria. Conclusions The results of this study suggest that E. hebecarpa extracts can be used to inhibit the planktonic and biofilm forms of these selected bacteria. PMID:27635214

  17. The transition between active and de-activated forms of NADH:ubiquinone oxidoreductase (Complex I) in the mitochondrial membrane of Neurospora crassa.

    PubMed Central

    Grivennikova, Vera G; Serebryanaya, Darya V; Isakova, Elena P; Belozerskaya, Tatyana A; Vinogradov, Andrei D

    2003-01-01

    The mammalian mitochondrial NADH:ubiquinone oxidoreductase (Complex I) has been shown to exist in two kinetically and structurally distinct slowly interconvertible forms, active (A) and de-activated (D) [Vinogradov and Grivennikova (2001) IUBMB Life 52, 129-134]. This work was undertaken to investigate the putative Complex I A-D transition in the mitochondrial membrane of the lower eukaryote Neurospora crassa and in plasma membrane of the prokaryote Paracoccus denitrificans, organisms that are eligible for molecular genetic manipulations. The potential interconversion between A and D forms was assessed by examination of the initial and steady-state rates of NADH oxidation catalysed by inside-out submitochondrial ( N. crassa ) and sub-bacterial ( P. denitrificans ) particles and their sensitivities to N -ethylmaleimide and Mg(2+). All diagnostic tests provide evidence that slow temperature- and turnover-dependent A-D transition is an explicit feature of eukaryotic N. crassa Complex I, whereas the phenomenon is not seen in the membranes of the prokaryote P. denitrificans. Significantly lower activation energy for A-to-D transition characterizes the N. crassa enzyme compared with that determined previously for the mammalian Complex I. Either a lag or a burst in the onset of the NADH oxidase assayed in the presence of Mg(2+) is seen when the reaction is initiated by the thermally de-activated or NADH-activated particles, whereas the delayed final activities of both preparations are the same. We conclude that continuous slow cycling between A and D forms occurs during the steady-state operation of Complex I in N. crassa mitochondria. PMID:12379145

  18. Sensitivity tests of the waste-form-alone design for the low-activity-waste disposal system

    SciTech Connect

    Fayer, M.J.; White, M.D.; Kincaid, C.T.

    1997-09-01

    Computer simulations were performed to assess the performance of the waste-form-alone (WFA) design for the low-activity-waste (LAW) disposal system. In FY 1997, PNNL performed additional simulations for Lockheed Martin Hanford Company (LMHC) to address specific questions about the disposal. LMHC manages the Glass Performance Assessment Project for DOE. The objectives of the additional simulations were to demonstrate the impact of grid resolution, diffusion, fracture flow within the waste form, and consumption of water by the waste form. It was assumed that the waste form would be the only part of the engineered disposal system that inhibits radionuclide release, referred to as WFA assumption. All calculations were performed with the latest version of the STOMP (Subsurface Transport Over Multiple Phases) simulator. Multiple simulations of the WFA disposal were performed to identify parameter and conceptual model sensitivities. The corrosion rate, recharge rate, well interception factor, hydraulic properties, and hydraulic and retardation models were shown to be important. Diffusion was shown to be important for the gravel model of glass but not the soil model. The impact of temperature changes was discussed and determined to be negligible. Water consumption during corrosion was evaluated and found to have a minimal effect on the dose calculations. Fracture flow within the glass was evaluated and found to have only a minimal effect on the dose calculations.

  19. An 11-kDa form of human immunodeficiency virus protease expressed in Escherichia coli is sufficient for enzymatic activity.

    PubMed Central

    Graves, M C; Lim, J J; Heimer, E P; Kramer, R A

    1988-01-01

    In order to define the protease domain of human immunodeficiency virus 1, various regions of the pol open reading frame were cloned and expressed in Escherichia coli. Antiserum directed against the conserved retroviral protease active site was used to identify pol precursor and processed species containing the presumed protease domain. The smallest product that accumulates is about 11 kDa as measured by NaDodSO4/PAGE. This size agrees with that predicted from the presence in this region of two Phe-Pro sequences, which is one of the cleavage sites recognized by HIV protease. DNA encoding only the predicted 11-kDa protein was cloned, bypassing the need for autoprocessing, and the protein was expressed to a high level in E. coli. This form is active as demonstrated by its ability to specifically cleave protease-deficient pol protein in vivo in E. coli. Extracts of E. coli containing the 11-kDa protease also process human immunodeficiency virus gag substrates in vitro. These results demonstrate that the 11-kDa protease is sufficient for enzymatic activity and are consistent with a major role for this form in virus maturation. Images PMID:3282230

  20. Spatial arrangement and functional role of α subunits of proteasome activator PA28 in hetero-oligomeric form

    SciTech Connect

    Sugiyama, Masaaki; Sahashi, Hiroki; Kurimoto, Eiji; Takata, Shin-ichi; Yagi, Hirokazu; Kanai, Keita; Sakata, Eri; Minami, Yasufumi; Tanaka, Keiji; Kato, Koichi

    2013-03-01

    Highlights: ► Homologous α and β subunits are alternatively arranged in the PA28 heptameric ring. ► The flexible loops of the three α subunits surround the site of substrate entry. ► The loops serve as gatekeepers that selectively hinder passage of longer peptides. - Abstract: A major form of proteasome activator PA28 is a heteroheptamer composed of interferon-γ-inducible α and β subunits, which share approximately 50% amino acid identity and possess distinct insert loops. This activator forms a complex with the 20S proteasome and thereby stimulates proteasomal degradation of peptides in an ATP-independent manner, giving rise to smaller antigenic peptides presented by major histocompatibility complex class I molecules. In this study, we performed biophysical and biochemical characterization of the structure and function of the PA28 hetero-oligomer. Deuteration-assisted small-angle neutron scattering demonstrated three α and four β subunits are alternately arranged in the heptameric ring. In this arrangement, PA28 loops surround the central pore of the heptameric ring (site for peptide entry). Activating the 20S proteasome with a PA28 mutant that lacked the α subunit loops cleaved model substrates longer than a nonapeptide with better efficiency when compared to wild-type PA28. Based on these data, we hypothesize that the flexible PA28 loops act as gatekeepers, which function to select the length of peptide substrates to be transported between the proteolytic chamber and the extra-proteasomal medium.

  1. Low-molecular-weight adiponectin is more closely associated with disease activity of rheumatoid arthritis than other adiponectin multimeric forms.

    PubMed

    Li, Ping; Yang, Li; Ma, Cui-Li; Liu, Bo; Zhang, Xin; Ding, Rui; Bi, Li-qi

    2015-06-01

    Adiponectin is divided into high-molecular-weight (HMW), medium-molecular-weight (MMW), and low-molecular-weight (LMW) forms. These forms differ not only in the number of adiponectin molecules but also in their biological activity. There are conflicting findings regarding the role of adiponectin in rheumatoid arthritis (RA). Moreover, few reports have described the relationships between serum adiponectin multimers levels and RA. Therefore, we examined the association of total adiponectin and its multimers with RA. Two study groups were examined: 180 recently diagnosed untreated RA patients with disease duration less than 1 year (RA group) and 160 age- and sex-matched control subjects (control group). RA-related factors, blood pressure, body mass index, glucose, complete lipid profile, and adiponectin multimers were measured. The levels of total adiponectin and each multimer of adiponectin were significantly lower in the RA than in the control (P < 0.01). Serum levels of total, HMW, MMW, and LMW were positively correlated with triglycerides levels and negatively correlated with the Disease Activity Score for 28 joints (DAS28). Multivariate regression analysis showed that total, HMW, and MMW adiponectin were independently associated with serum triglycerides level. LMW adiponectin was independently correlated with serum triglycerides level and DAS28. The decreased LMW adiponectin levels may be associated with disease activity of RA.

  2. Anti-Biofilm and Immunomodulatory Activities of Peptides That Inhibit Biofilms Formed by Pathogens Isolated from Cystic Fibrosis Patients.

    PubMed

    de la Fuente-Núñez, César; Mansour, Sarah C; Wang, Zhejun; Jiang, Lucy; Breidenstein, Elena B M; Elliott, Melissa; Reffuveille, Fany; Speert, David P; Reckseidler-Zenteno, Shauna L; Shen, Ya; Haapasalo, Markus; Hancock, Robert E W

    2014-01-01

    Cystic fibrosis (CF) patients often acquire chronic respiratory tract infections due to Pseudomonas aeruginosa and Burkholderia cepacia complex (Bcc) species. In the CF lung, these bacteria grow as multicellular aggregates termed biofilms. Biofilms demonstrate increased (adaptive) resistance to conventional antibiotics, and there are currently no available biofilm-specific therapies. Using plastic adherent, hydroxyapatite and flow cell biofilm models coupled with confocal and scanning electron microscopy, it was demonstrated that an anti-biofilm peptide 1018 prevented biofilm formation, eradicated mature biofilms and killed biofilms formed by a wide range of P. aeruginosa and B. cenocepacia clinical isolates. New peptide derivatives were designed that, compared to their parent peptide 1018, showed similar or decreased anti-biofilm activity against P. aeruginosa biofilms, but increased activity against biofilms formed by the Gram-positive bacterium methicillin resistant Staphylococcus aureus. In addition, some of these new peptide derivatives retained the immunomodulatory activity of 1018 since they induced the production of the chemokine monocyte chemotactic protein-1 (MCP-1) and suppressed lipopolysaccharide-mediated tumor necrosis factor-α (TNF-α) production by human peripheral blood mononuclear cells (PBMC) and were non-toxic towards these cells. Peptide 1018 and its derivatives provide promising leads for the treatment of chronic biofilm infections and hyperinflammatory lung disease in CF patients.

  3. Adhesive Properties and Acid-Forming Activity of Lactobacilli and Streptococci Under Inhibitory Substances, Such as Nitrates.

    PubMed

    Hakobyan, L; Harutyunyan, K; Harutyunyan, N; Melik-Andreasyan, G; Trchounian, A

    2016-06-01

    One of the main requirements for probiotics is their ability to survive during passage through gastrointestinal tract and to maintain their activity at different adverse conditions. The aim of the study was to look for the strains of lactobacilli and streptococci with high adhesive properties even affected by inhibitory substances, such as nitrates (NO3 (-)). To study the adhesion properties hemagglutination reaction of bacterial cells with red blood cells of different animals and humans was used. The acid formation ability of bacteria was determined by the method of titration after 7 days of incubation in the sterile milk. These properties were investigated at different concentrations of NO3 (-). The high concentration (mostly ≥2.0 %) NO3 (-) inhibited the growth of both lactobacilli and streptococci, but compared with streptococcal cultures lactobacilli, especially Lactobacillus acidophilus Ep 317/402, have shown more stability and higher adhesive properties. In addition, the concentrations of NO3 (-) of 0.5-2.0 % decreased the acid-forming activity of the strains, but even under these conditions they coagulated milk and, in comparison to control, formed low acidity in milk. Thus, the L. acidophilus Ep 317/402 with high adhesive properties has demonstrated a higher activity of NO3 (-) transformation.

  4. Cytotoxic activities of Leptospira interrogans hemolysin SphH as a pore-forming protein on mammalian cells.

    PubMed

    Lee, Seoung Hoon; Kim, Sangduk; Park, Seung Chul; Kim, Min Ja

    2002-01-01

    Leptospirosis is a spirochetal zoonosis that causes an acute febrile systemic illness in humans. Leptospira sp. hemolysins have been shown to be virulence factors for the pathogenesis of leptospirosis. Previously, we cloned a hemolysin SphH of Leptospira interrogans serovar lai, a homologue of L. borgpetersenii sphingomyelinase (SphA), from a genomic library (S. H. Lee, K. A. Kim, Y. K. Kim, I. W. Seong, M. J. Kim, and Y. J. Lee, Gene 254:19-28, 2000). Escherichia coli lysate harboring the sphH showed high hemolytic activities on sheep erythrocytes. However, it neither showed sphingomyelinase nor phospholipase activities, in contrast to SphA which was known to have sphingomyelinase activity. Interestingly, the SphH-mediated hemolysis on erythrocytes was osmotically protected by PEG 5000, suggesting that the SphH might have caused pore formation on the erythrocyte membrane. In the present study, we have prepared the Leptospira hemolysin SphH and investigated its hemolytic and cytotoxic activities on mammalian cells. SphH was shown to be a pore-forming protein on several mammalian cells: When treated with the SphH, the sheep erythrocyte membranes formed pores, which were morphologically confirmed by transmission electron microscopy. Furthermore, the SphH-mediated cytotoxicities on mammalian cells were demonstrated by the release of LDH and by inverted microscopic examinations. Finally, the immune serum against the full-length hemolysin could effectively neutralize the SphH-mediated hemolytic and cytotoxic activities. In conclusion, these results suggest that the virulence of Leptospira SphH was due to the pore formation on mammalian cell membranes.

  5. Forms of selenium affect its transport, uptake and glutathione peroxidase activity in the Caco-2 cell model.

    PubMed

    Wang, Yanbo; Fu, Linglin

    2012-10-01

    The experiment was designed to investigate the effect of selenium (Se) chemical forms (sodium selenite, selenium nanoparticle [nano-Se] and selenomethionine) on the transport, uptake and glutathione peroxidase (GSH-Px) activity in the Caco-2 cell model. The transport and uptake of different forms of Se (0.1 μmol l(-1)) across the Caco-2 cell monolayer were carried out in two directions (apical [AP] to basolateral [BL] and BL to AP) for 2 h, respectively, and the apparent permeability coefficient (P(app)), transport efficiency and uptake efficiency were all calculated. In the present study, the transport and uptake of three forms of Se were time-dependent both in AP to BL and BL to AP directions. By the end of 2 h, the transport efficiencies of selenomethionine and nano-Se were higher than that of sodium selenite (P<0.05). The highest uptake efficiency (P<0.05) was observed in cells treated with nano-Se and significant difference (P<0.05) was also observed between the cells incubated with sodium selenite and selenomethionine. As for the P(app), sodium selenite (P<0.05) had the lowest values compared with that of selenomethionine and nano-Se, in both AP-BL and BL-AP. However, no significant differences were observed in GSH-Px activities. These results indicated that the efficiency of Se in the Caco-2 cells varied with its chemical forms, which might be associated with the differences in Se transport and uptake.

  6. Paradoxical antifungal activity and structural observations in biofilms formed by echinocandin-resistant Candida albicans clinical isolates.

    PubMed

    Walraven, Carla J; Bernardo, Stella M; Wiederhold, Nathan P; Lee, Samuel A

    2014-02-01

    Echinocandin-resistant clinical isolates of Candida albicans have been reported, and key-hot spot mutations in the FKS1 gene, which encodes a major glucan synthase subunit, have been identified in these (caspofungin-resistant [CAS-R]) strains. Although these mutations result in phenotypic resistance to echinocandins in planktonic cells, there is little data on antifungal susceptibilities of CAS-R C. albicans strains within biofilms. Thus, we analyzed biofilms formed by 12 C. albicans CAS-R clinical strains in which we previously identified FKS1 hot-spot mutations and compared the sessile antifungal and paradoxical activity of anidulafungin (ANID), caspofungin (CAS), and micafungin (MICA). Biofilms were formed in a 96-well static microplate model and assayed using both tetrazolium-salt reduction and crystal violet assays, as well as examination by scanning electron microscopy. We first sought to assess biofilm formation and structure in these fks1 mutants and found that the biofilm mass and metabolic activities were reduced in most of the fks1 mutants as compared with reference strain SC5314. Structural analyses revealed that the fks1 mutant biofilms were generally less dense and had a clear predominance of yeast and pseudohyphae, with unusual "pit"-like cell surface structures. We also noted that sessile minimum inhibitory concentrations (MICs) to ANID, CAS, and MICA were higher than planktonic MICs of all but one strain. The majority of strains demonstrated a paradoxical effect (PE) to particular echinocandins, in either planktonic or sessile forms. Overall, biofilms formed by echinocandin-resistant clinical isolates demonstrated varied PEs to echinocandins and were structurally characterized by a preponderance of yeast, pseudohyphae, and pit-like structures.

  7. GOODS-HERSCHEL: SEPARATING HIGH-REDSHIFT ACTIVE GALACTIC NUCLEI AND STAR-FORMING GALAXIES USING INFRARED COLOR DIAGNOSTICS

    SciTech Connect

    Kirkpatrick, Allison; Pope, Alexandra; Charmandaris, Vassilis; Daddi, Emmanuele; Elbaz, David; Pannella, Maurilio; Aussel, Herve; Dasyra, Kalliopi; Leiton, Roger; Scott, Douglas; Magnelli, Benjamin; Popesso, Paola; Altieri, Bruno; Coia, Daniela; Valtchanov, Ivan; Dannerbauer, Helmut; Dickinson, Mark; Kartaltepe, Jeyhan; Magdis, Georgios

    2013-02-15

    We have compiled a large sample of 151 high-redshift (z = 0.5-4) galaxies selected at 24 {mu}m (S {sub 24} > 100 {mu}Jy) in the GOODS-N and ECDFS fields for which we have deep Spitzer IRS spectroscopy, allowing us to decompose the mid-infrared spectrum into contributions from star formation and activity in the galactic nuclei. In addition, we have a wealth of photometric data from Spitzer IRAC/MIPS and Herschel PACS/SPIRE. We explore how effective different infrared color combinations are at separating our mid-IR spectroscopically determined active galactic nuclei from our star-forming galaxies. We look in depth at existing IRAC color diagnostics, and we explore new color-color diagnostics combining mid-IR, far-IR, and near-IR photometry, since these combinations provide the most detail about the shape of a source's IR spectrum. An added benefit of using a color that combines far-IR and mid-IR photometry is that it is indicative of the power source driving the IR luminosity. For our data set, the optimal color selections are S {sub 250}/S {sub 24} versus S {sub 8}/S {sub 3.6} and S {sub 100}/S {sub 24} versus S {sub 8}/S {sub 3.6}; both diagnostics have {approx}10% contamination rate in the regions occupied primarily by star-forming galaxies and active galactic nuclei, respectively. Based on the low contamination rate, these two new IR color-color diagnostics are ideal for estimating both the mid-IR power source of a galaxy when spectroscopy is unavailable and the dominant power source contributing to the IR luminosity. In the absence of far-IR data, we present color diagnostics using the Wide-field Infrared Survey Explorer mid-IR bands which can efficiently select out high-z (z {approx} 2) star-forming galaxies.

  8. Evaluation of antinociceptive activity of nanoliposome-encapsulated and free-form diclofenac in rats and mice.

    PubMed

    Goh, Jun Zheng; Tang, Sook Nai; Chiong, Hoe Siong; Yong, Yoke Keong; Zuraini, Ahmad; Hakim, Muhammad Nazrul

    2015-01-01

    Diclofenac is a nonsteroidal anti-inflammatory drug (NSAID) that exhibits anti-inflammatory, antinociceptive, and antipyretic activities. Liposomes have been shown to improve the therapeutic efficacy of encapsulated drugs. The present study was conducted to compare the antinociceptive properties between liposome-encapsulated and free-form diclofenac in vivo via different nociceptive assay models. Liposome-encapsulated diclofenac was prepared using the commercialized proliposome method. Antinociceptive effects of liposome-encapsulated and free-form diclofenac were evaluated using formalin test, acetic acid-induced abdominal writhing test, Randall-Selitto paw pressure test, and plantar test. The results of the writhing test showed a significant reduction of abdominal constriction in all treatment groups in a dose-dependent manner. The 20 mg/kg liposome-encapsulated diclofenac demonstrated the highest antinociceptive effect at 78.97% compared with 55.89% in the free-form group at equivalent dosage. Both liposome-encapsulated and free-form diclofenac produced significant results in the late phase of formalin assay at a dose of 20 mg/kg, with antinociception percentages of 78.84% and 60.71%, respectively. Significant results of antinociception were also observed in both hyperalgesia assays. For Randall-Sellito assay, the highest antinociception effect of 71.38% was achieved with 20 mg/kg liposome-encapsulated diclofenac, while the lowest antinociceptive effect of 17.32% was recorded with 0 mg/kg liposome formulation, whereas in the plantar test, the highest antinociceptive effect was achieved at 56.7% with 20 mg/kg liposome-encapsulated diclofenac, and the lowest effect was shown with 0 mg/kg liposome formulation of 8.89%. The present study suggests that liposome-encapsulated diclofenac exhibits higher antinociceptive efficacy in a dose-dependent manner in comparison with free-form diclofenac.

  9. Evaluation of antinociceptive activity of nanoliposome-encapsulated and free-form diclofenac in rats and mice

    PubMed Central

    Goh, Jun Zheng; Tang, Sook Nai; Chiong, Hoe Siong; Yong, Yoke Keong; Zuraini, Ahmad; Hakim, Muhammad Nazrul

    2015-01-01

    Diclofenac is a nonsteroidal anti-inflammatory drug (NSAID) that exhibits anti-inflammatory, antinociceptive, and antipyretic activities. Liposomes have been shown to improve the therapeutic efficacy of encapsulated drugs. The present study was conducted to compare the antinociceptive properties between liposome-encapsulated and free-form diclofenac in vivo via different nociceptive assay models. Liposome-encapsulated diclofenac was prepared using the commercialized proliposome method. Antinociceptive effects of liposome-encapsulated and free-form diclofenac were evaluated using formalin test, acetic acid-induced abdominal writhing test, Randall–Selitto paw pressure test, and plantar test. The results of the writhing test showed a significant reduction of abdominal constriction in all treatment groups in a dose-dependent manner. The 20 mg/kg liposome-encapsulated diclofenac demonstrated the highest antinociceptive effect at 78.97% compared with 55.89% in the free-form group at equivalent dosage. Both liposome-encapsulated and free-form diclofenac produced significant results in the late phase of formalin assay at a dose of 20 mg/kg, with antinociception percentages of 78.84% and 60.71%, respectively. Significant results of antinociception were also observed in both hyperalgesia assays. For Randall–Sellito assay, the highest antinociception effect of 71.38% was achieved with 20 mg/kg liposome-encapsulated diclofenac, while the lowest antinociceptive effect of 17.32% was recorded with 0 mg/kg liposome formulation, whereas in the plantar test, the highest antinociceptive effect was achieved at 56.7% with 20 mg/kg liposome-encapsulated diclofenac, and the lowest effect was shown with 0 mg/kg liposome formulation of 8.89%. The present study suggests that liposome-encapsulated diclofenac exhibits higher antinociceptive efficacy in a dose-dependent manner in comparison with free-form diclofenac. PMID:25678786

  10. PROCESS OF TREATING OR FORMING AN INSOLUBLE PLUTONIUM PRECIPITATE IN THE PRESENCE OF AN ORGANIC ACTIVE AGENT

    DOEpatents

    Balthis, J.H.

    1961-07-18

    Carrier precipitation processes for the separation of plutonium from fission products are described. In a process in which an insoluble precipitate is formed in a solution containing plutonium and fission products under conditions whereby plutonium is carried by the precipitate, and the precipitate is then separated from the remaining solution, an organic surface active agent is added to the mixture of precipitate and solution prior to separation of the precipitate from the supernatant solution, thereby improving the degree of separation of the precipitate from the solution.

  11. Purification in an active form of the phage phi 29 protein p4 that controls the viral late transcription.

    PubMed Central

    Barthelemy, I; Lázaro, J M; Méndez, E; Mellado, R P; Salas, M

    1987-01-01

    The phage phi 29 protein p4, that controls viral late transcription, was highly purified from Escherichia coli cells harbouring a gene 4-containing plasmid. This protein, representing about 6% of the total cellular protein, was obtained in a highly purified form. The protein was characterized as p4 by amino acid analysis and NH2-terminal sequence determination. The purified protein was active in an in vitro transcription assay, allowing specific initiation of transcription at the phi 29 A3 late promoter in the presence of Bacillus subtilis sigma 43-RNA polymerase holoenzyme. Images PMID:3671066

  12. Auxin efflux by PIN-FORMED proteins is activated by two different protein kinases, D6 PROTEIN KINASE and PINOID.

    PubMed

    Zourelidou, Melina; Absmanner, Birgit; Weller, Benjamin; Barbosa, Inês C R; Willige, Björn C; Fastner, Astrid; Streit, Verena; Port, Sarah A; Colcombet, Jean; de la Fuente van Bentem, Sergio; Hirt, Heribert; Kuster, Bernhard; Schulze, Waltraud X; Hammes, Ulrich Z; Schwechheimer, Claus

    2014-06-19

    The development and morphology of vascular plants is critically determined by synthesis and proper distribution of the phytohormone auxin. The directed cell-to-cell distribution of auxin is achieved through a system of auxin influx and efflux transporters. PIN-FORMED (PIN) proteins are proposed auxin efflux transporters, and auxin fluxes can seemingly be predicted based on the--in many cells--asymmetric plasma membrane distribution of PINs. Here, we show in a heterologous Xenopus oocyte system as well as in Arabidopsis thaliana inflorescence stems that PIN-mediated auxin transport is directly activated by D6 PROTEIN KINASE (D6PK) and PINOID (PID)/WAG kinases of the Arabidopsis AGCVIII kinase family. At the same time, we reveal that D6PKs and PID have differential phosphosite preferences. Our study suggests that PIN activation by protein kinases is a crucial component of auxin transport control that must be taken into account to understand auxin distribution within the plant.

  13. Modification biological activity of S and R forms of Proteus mirabilis and Burkholderia cepacia lipopolysaccharides by carrageenans.

    PubMed

    Arabski, Michał; Barabanova, Anna; Gałczyńska, Katarzyna; Węgierek-Ciuk, Aneta; Dzidowska, Kamila; Augustyniak, Daria; Drulis-Kawa, Zuzanna; Lankoff, Anna; Yermak, Irina; Molinaro, Antonio; Kaca, Wiesław

    2016-09-20

    The modification of biological features of S and R forms of Proteus mirabilis and Burkholderia cepacia LPS by kappa/iota and kappa/beta carrageenans was shown in Limulus activation test, ELISA, human complement activation and apoptotic assay. The role of positively charged substituent Ara4N in lipid A was evaluated as a suspected major domain for interactions with sulphate groups of carrageenans.The experiments obtained by three serological methods indicated that not only lipid A part of LPS but also polysaccharide elements such as core and O-specific chain are involved in interaction with carrageenes. Carrageenans turned out to be non-cytotoxic for A549 cells and were able to inhibit the apoptotic effect caused by lipid A of P. mirabilis and B. cepacia.

  14. Selection of suitable detergents for obtaining an active dengue protease in its natural form from E. coli.

    PubMed

    Liew, Lynette Sin Yee; Lee, Michelle Yueqi; Wong, Ying Lei; Cheng, Jinting; Li, Qingxin; Kang, CongBao

    2016-05-01

    Dengue protease is a two-component enzyme and is an important drug target against dengue virus. The protease activity and protein stability of dengue nonstructural protein 3 (NS3) require a co-factor region from a four-span membrane protein NS2B. A natural form of dengue protease containing full-length NS2B and NS3 protease domain NS2BFL-NS3pro will be useful for dengue drug discovery. In current study, detergents that can be used for protease purification were tested. Using a water soluble protease construct, 39 detergents were selected for both NS2B and NS2BFL-NS3pro purification. The results showed that 18 detergents were able to sustain the activity of the natural dengue protease and 11 detergents could be used for NS2B purification. The results obtained in this study will be useful for biochemical and biophysical studies on dengue protease.

  15. Structural investigation of zymogenic and activated forms of human blood coagulation factor VIII: a computational molecular dynamics study

    PubMed Central

    2010-01-01

    Background Human blood coagulation factor VIII (fVIII) is a large plasma glycoprotein with sequential domain arrangement in the order A1-a1-A2-a2-B-a3-A3-C1-C2. The A1, A2 and A3 domains are interconnected by long linker peptides (a1, a2 and a3) that possess the activation sites. Proteolysis of fVIII zymogen by thrombin or factor Xa results in the generation of the activated form (fVIIIa) which serves as a critical co-factor for factor IXa (fIXa) enzyme in the intrinsic coagulation pathway. Results In our efforts to elucidate the structural differences between fVIII and fVIIIa, we developed the solution structural models of both forms, starting from an incomplete 3.7 Å X-ray crystal structure of fVIII zymogen, using explicit solvent MD simulations. The full assembly of B-domainless single-chain fVIII was built between the A1-A2 (Ala1-Arg740) and A3-C1-C2 (Ser1669-Tyr2332) domains. The structural dynamics of fVIII and fVIIIa, simulated for over 70 ns of time scale, enabled us to evaluate the integral motions of the multi-domain assembly of the co-factor and the possible coordination pattern of the functionally important calcium and copper ion binding in the protein. Conclusions MD simulations predicted that the acidic linker peptide (a1) between the A1 and A2 domains is largely flexible and appears to mask the exposure of putative fIXa enzyme binding loop (Tyr555-Asp569) region in the A2 domain. The simulation of fVIIIa, generated from the zymogen structure, predicted that the linker peptide (a1) undergoes significant conformational reorganization upon activation by relocating completely to the A1-domain. The conformational transition led to the exposure of the Tyr555-Asp569 loop and the surrounding region in the A2 domain. While the proposed linker peptide conformation is predictive in nature and warrants further experimental validation, the observed conformational differences between the zymogen and activated forms may explain and support the large body of

  16. Structural and biochemical studies on Vibrio cholerae Hsp31 reveals a novel dimeric form and Glutathione-independent Glyoxalase activity

    PubMed Central

    Dey, Sanjay

    2017-01-01

    Vibrio cholerae experiences a highly hostile environment at human intestine which triggers the induction of various heat shock genes. The hchA gene product of V. cholerae O395, referred to a hypothetical intracellular protease/amidase VcHsp31, is one such stress-inducible homodimeric protein. Our current study demonstrates that VcHsp31 is endowed with molecular chaperone, amidopeptidase and robust methylglyoxalase activities. Through site directed mutagenesis coupled with biochemical assays on VcHsp31, we have confirmed the role of residues in the vicinity of the active site towards amidopeptidase and methylglyoxalase activities. VcHsp31 suppresses the aggregation of insulin in vitro in a dose dependent manner. Through crystal structures of VcHsp31 and its mutants, grown at various temperatures, we demonstrate that VcHsp31 acquires two (Type-I and Type-II) dimeric forms. Type-I dimer is similar to EcHsp31 where two VcHsp31 monomers associate in eclipsed manner through several intersubunit hydrogen bonds involving their P-domains. Type-II dimer is a novel dimeric organization, where some of the intersubunit hydrogen bonds are abrogated and each monomer swings out in the opposite directions centering at their P-domains, like twisting of wet cloth. Normal mode analysis (NMA) of Type-I dimer shows similar movement of the individual monomers. Upon swinging, a dimeric surface of ~400Å2, mostly hydrophobic in nature, is uncovered which might bind partially unfolded protein substrates. We propose that, in solution, VcHsp31 remains as an equilibrium mixture of both the dimers. With increase in temperature, transformation to Type-II form having more exposed hydrophobic surface, occurs progressively accounting for the temperature dependent increase of chaperone activity of VcHsp31. PMID:28235098

  17. Specifically Formed Corona on Silica Nanoparticles Enhances Transforming Growth Factor β1 Activity in Triggering Lung Fibrosis.

    PubMed

    Wang, Zhenzhen; Wang, Chunming; Liu, Shang; He, Wei; Wang, Lintao; Gan, JingJing; Huang, Zhen; Wang, Zhenheng; Wei, Haoyang; Zhang, Junfeng; Dong, Lei

    2017-02-28

    A corona is a layer of macromolecules formed on a nanoparticle surface in vivo. It can substantially change the biological identity of nanomaterials and possibly trigger adverse responses from the body tissues. Dissecting the role of the corona in the development of a particular disease may provide profound insights for understanding toxicity of nanomaterials in general. In our present study, we explored the capability of different silica nanoparticles (SiNPs) to induce silicosis in the mouse lung and analyzed the composition of coronas formed on these particles. We found that SiNPs of certain size and surface chemistry could specifically recruit transforming growth factor β1 (TGF-β1) into their corona, which subsequently induces the development of lung fibrosis. Once embedded into the corona on SiNPs, TGF-β1 was remarkably more stable than in its free form, and its fibrosis-triggering activity was significantly prolonged. Our study meaningfully demonstrates that a specific corona component on a certain nanoparticle could initiate a particular pathogenic process in a clinically relevant disease model. Our findings may shed light on the understanding of molecular mechanisms of human health risks correlated with exposure to small-scale substances.

  18. Study on the effects of nylon-chitosan-blended membranes on the spheroid-forming activity of human melanocytes.

    PubMed

    Lin, Sung-Jan; Hsiao, Wen-Chu; Jee, Shiou-Hwa; Yu, Hsin-Su; Tsai, Tsen-Fang; Lai, Juin-Yih; Young, Tai-Horng

    2006-10-01

    Though reported limitedly in tissue engineering, modification of cellular functions can be achieved by culturing them into multicellular spheroids. We have shown melanocytes form spheroids on chitosan surface. However, how biomaterials promote spheroid formation has never been systemically investigated. In this work, nylon, which inhibits melanocyte spheroid formation, and chitosan, which promotes melanocyte spheroid formation, are used to prepare nylon/chitosan-blended membranes. Membranes composed of pure nylon, pure chitosan and various ratios of nylon and chitosan are employed to examine their effects on spheroid formation. Melanocytes show better adhesion to nylon membranes than that to chitosan membranes. In blended membranes, as more nylon is incorporated, cell adhesion increases and the trend for spheroid formation decreases. Melanocytes can only form spheroids on membranes with poorer cell adhesion. Examining the surface of the blended membranes shows phase separation of nylon and chitosan. As nylon content increases, the nylon phase on the membrane surface increases and thereby enhances cell adhesion. The opposite trend for cell adhesion and spheroid formation substantiates our hypothesis of spheroid formation on biomaterials: a balance between cell-substrate interaction and cell-cell interaction. The decrease in cell-substrate interaction tilts the balance to a state more favorable for spheroid formation. Our work can serve as a model to investigate the relative strengths of cell-cell and cell-substrate interactions and also pave way to design blended membranes with desired physical properties while preserving the spheroid-forming activity.

  19. Activated and nonactivated forms of zinc powder: Reactivity toward chlorocarbons in water and AFM studies of surface morphologies

    SciTech Connect

    Boronina, T.N.; Lagadic, I.; Klabunde, K.J.; Sergeev, G.B.

    1998-09-01

    This work was carried out with the purpose of developing effective reagents for decontamination of groundwater contaminated with chlorocarbons. Zinc metal as a reducing agent for carbon tetrachloride (CT), chloroform (Chl), and methylene chloride (MC) in aqueous solution has been studied in some detail, especially regarding activated forms of the metal. Chlorocarbon concentrations were monitored at certain time intervals by gas chromatography/mass spectrometry (GC/MS) analysis of the headspace and water phase. Reaction mixture headspace was additionally studied by a GC/headspace analysis system to detect the formation of hydrocarbons. Chloroform, methylene chloride, methyl chloride, methane, and acetylene were found to be products from CT reduction. For methylene chloride reduction, traces of cis and trans-1,2-dichloroethene (DCE) were also found. Activated by cryo or mechanical treatment, metallic zinc caused an increase in CT dechlorination rate and conversion into methane. After the first 2.5 h, more than 20% of CT was converted into methane by cryochemically activated zinc in comparison to 1.2% by conventional zinc dust. Furthermore, CT reduction by activated zinc caused the formation of DCEs and TCE. Pathways are proposed to account for the observed methane/methylene chloride ratio and DCEs and TCE formation that include sequential reductive dechlorination through organometallic and carbonoid species on the Zn surface.

  20. Production of biologically active recombinant goose FSH in a single chain form with a CTP linker sequence.

    PubMed

    Li, Hui; Zhu, Huanxi; Qin, Qinming; Lei, Mingming; Shi, Zhendan

    2017-02-01

    FSH is a glycoprotein hormone secreted by the pituitary gland that is essential for gonadal development and reproductive function. In avian reproduction study, especially in avian reproduction hormone study, it is hindered by the lack of biologically active FSH. In order to overcome this shortcoming, we prepared recombinant goose FSH as a single chain molecule and tested its biological activities in the present study. Coding sequences for mature peptides of goose FSH α and β subunits were amplified from goose pituitary cDNA. A chimeric gene containing α and β subunit sequences linked by the hCG carboxyl terminal peptide coding sequence was constructed. The recombinant gene was inserted into the pcDNA3.1-Fc eukaryotic expression vector to form pcDNA-Fc-gFSHβ-CTP-α and then transfected into 293-F cells. A recombinant, single chain goose FSH was expressed and verified by SDS-PAGE and western blot analysis, and was purified using Protein A agarose affinity and gel filtration chromatography. Biological activity analysis results showed that the recombinant, chimeric goose FSH possesses the function of stimulating estradiol secretion and cell proliferation, in cultured chicken granulosa cells. These results indicated that bioactive, recombinant goose FSH has been successfully prepared in vitro. The recombinant goose FSH will have the potential of being used as a research tool for studying avian reproductive activities, and as a standard for developing avian FSH bioassays.

  1. Trypanosoma cruzi: antiprotozoal activity of parthenolide obtained from Tanacetum parthenium (L.) Schultz Bip. (Asteraceae, Compositae) against epimastigote and amastigote forms.

    PubMed

    Izumi, Erika; Morello, Luis Gustavo; Ueda-Nakamura, Tânia; Yamada-Ogatta, Sueli Fumie; Filho, Benedito Prado Dias; Cortez, Diógenes Aparício Garcia; Ferreira, Izabel Cristina Piloto; Morgado-Díaz, José Andrés; Nakamura, Celso Vataru

    2008-03-01

    This study reports the activity of crude extracts, fractions and parthenolide (pure compound) obtained from Tanacetum parthenium against two forms of the parasite Trypanosoma cruzi. Feverfew is a traditional herbal medicine that has been used for the treatment of migraine, fever and arthritis. Activity against epimastigote forms was observed for crude extracts, fractions and parthenolide, and a progressive increase in the antitrypanosomal effect was observed in the course of the purification process. The pure compound showed IC50/96h and IC90/96h of 0.5 microg/ml and 1.25 microg/ml, respectively. The cytotoxic effect of parthenolide in LLMCK2 cells was 3.2 microg/ml (CC50/96h) and the selectivity index was 6.4. No hemolysis was detected for the pure compound. The internalization index of T. cruzi in LLMCK2 cells was reduced almost 51% at the concentration of 2 microg/ml of parthenolide, and 96.6% at 4 microg/ml. Scanning and transmission electron microscopy permitted observation of morphological modifications and ultrastructural alterations.

  2. A Staphylococcus aureus Pore-Forming Toxin Subverts the Activity of ADAM10 to Cause Lethal Infection

    PubMed Central

    Inoshima, Ichiro; Inoshima, Naoko; Wilke, Georgia; Powers, Michael; Frank, Karen; Wang, Yang; Wardenburg, Juliane Bubeck

    2011-01-01

    Staphylococcus aureus is a major cause of human disease, responsible for half a million infections and approximately 20,000 deaths per year in the United States alone 1,2. This pathogen secretes α-hemolysin, a pore-forming cytotoxin that contributes to the pathogenesis of pneumonia 3–5. α-hemolysin injures epithelial cells by interacting with its receptor, the zinc-dependent metalloprotease ADAM10 6. We show that mice harboring a conditional disruption of the Adam10 gene in lung epithelium are resistant to lethal pneumonia. Investigation of the molecular mechanism of toxin-receptor function revealed that α-hemolysin upregulates ADAM10 metalloprotease activity in alveolar epithelial cells, resulting in cleavage of the adherens junction protein E-cadherin. Cleavage is associated with disruption of epithelial barrier function, contributing to the pathogenesis of lethal acute lung injury. A metalloprotease inhibitor of ADAM10 prevents E-cadherin cleavage; similarly, E-cadherin proteolysis and barrier disruption is attenuated in ADAM10 knockout mice. Together, these data attest to the function of ADAM10 as the cellular receptor for α-hemolysin. The observation that Hla can usurp the metalloprotease activity of its receptor reveals a novel mechanism of pore-forming cytotoxin action in which pathologic insults are not solely the result of irreversible membrane injury, and defines ADAM10 inhibition as a strategy for disease modification. PMID:21926978

  3. Non-dimensionalised closed-form parametric analysis of semi-active vehicle suspensions using a quarter-car model

    NASA Astrophysics Data System (ADS)

    Ahmadian, Mehdi; Blanchard, Emmanuel

    2011-02-01

    This article provides a non-dimensionalised closed-form analysis of semi-active vehicle suspensions, using a quarter-car model. The derivation of the closed-form solutions for three indices that can be used for ride comfort, vehicle handling, and stability are presented based on non-dimensionalised suspension parameters. The behaviour of semi-active vehicle suspensions is evaluated using skyhook, groundhook, and hybrid control policies, and compared with passive suspensions. The relationship between vibration isolation, suspension deflection, and road holding is studied, using three performance indices based on the mean square of the sprung mass acceleration, rattle space, and tyre deflection, respectively. The results of the study indicate that the hybrid control policy yields significantly better comfort than a passive suspension, without reducing the road-holding quality or increasing the suspension displacement for typical passenger cars. The results also indicate that for typical passenger cars, the hybrid control policy results in a better compromise between comfort, road holding and suspension travel requirements than both the skyhook and groundhook control methods.

  4. An Early and Robust Activation of Caspases Heads Cells for a Regulated Form of Necrotic-like Cell Death*

    PubMed Central

    Garcia-Belinchón, Mercè; Sánchez-Osuna, María; Martínez-Escardó, Laura; Granados-Colomina, Carla; Pascual-Guiral, Sònia; Iglesias-Guimarais, Victoria; Casanelles, Elisenda; Ribas, Judit; Yuste, Victor J.

    2015-01-01

    Apoptosis is triggered by the activation of caspases and characterized by chromatin condensation and nuclear fragmentation (type II nuclear morphology). Necrosis is depicted by a gain in cell volume (oncosis), swelling of organelles, plasma membrane leakage, and subsequent loss of intracellular contents. Although considered as different cell death entities, there is an overlap between apoptosis and necrosis. In this sense, mounting evidence suggests that both processes can be morphological expressions of a common biochemical network known as “apoptosis-necrosis continuum.” To gain insight into the events driving the apoptosis-necrosis continuum, apoptotically proficient cells were screened facing several apoptotic inducers for the absence of type II apoptotic nuclear morphologies. Chelerythrine was selected for further studies based on its cytotoxicity and the lack of apoptotic nuclear alterations. Chelerythrine triggered an early plasma membrane leakage without condensed chromatin aggregates. Ultrastructural analysis revealed that chelerythrine-mediated cytotoxicity was compatible with a necrotic-like type of cell death. Biochemically, chelerythrine induced the activation of caspases. Moreover, the inhibition of caspases prevented chelerythrine-triggered necrotic-like cell death. Compared with staurosporine, chelerythrine induced stronger caspase activation detectable at earlier times. After using a battery of chemicals, we found that high concentrations of thiolic antioxidants fully prevented chelerythrine-driven caspase activation and necrotic-like cell death. Lower amounts of thiolic antioxidants partially prevented chelerythrine-mediated cytotoxicity and allowed cells to display type II apoptotic nuclear morphology correlating with a delay in caspase-3 activation. Altogether, these data support that an early and pronounced activation of caspases can drive cells to undergo a form of necrotic-like regulated cell death. PMID:26124276

  5. An Early and Robust Activation of Caspases Heads Cells for a Regulated Form of Necrotic-like Cell Death.

    PubMed

    Garcia-Belinchón, Mercè; Sánchez-Osuna, María; Martínez-Escardó, Laura; Granados-Colomina, Carla; Pascual-Guiral, Sònia; Iglesias-Guimarais, Victoria; Casanelles, Elisenda; Ribas, Judit; Yuste, Victor J

    2015-08-21

    Apoptosis is triggered by the activation of caspases and characterized by chromatin condensation and nuclear fragmentation (type II nuclear morphology). Necrosis is depicted by a gain in cell volume (oncosis), swelling of organelles, plasma membrane leakage, and subsequent loss of intracellular contents. Although considered as different cell death entities, there is an overlap between apoptosis and necrosis. In this sense, mounting evidence suggests that both processes can be morphological expressions of a common biochemical network known as "apoptosis-necrosis continuum." To gain insight into the events driving the apoptosis-necrosis continuum, apoptotically proficient cells were screened facing several apoptotic inducers for the absence of type II apoptotic nuclear morphologies. Chelerythrine was selected for further studies based on its cytotoxicity and the lack of apoptotic nuclear alterations. Chelerythrine triggered an early plasma membrane leakage without condensed chromatin aggregates. Ultrastructural analysis revealed that chelerythrine-mediated cytotoxicity was compatible with a necrotic-like type of cell death. Biochemically, chelerythrine induced the activation of caspases. Moreover, the inhibition of caspases prevented chelerythrine-triggered necrotic-like cell death. Compared with staurosporine, chelerythrine induced stronger caspase activation detectable at earlier times. After using a battery of chemicals, we found that high concentrations of thiolic antioxidants fully prevented chelerythrine-driven caspase activation and necrotic-like cell death. Lower amounts of thiolic antioxidants partially prevented chelerythrine-mediated cytotoxicity and allowed cells to display type II apoptotic nuclear morphology correlating with a delay in caspase-3 activation. Altogether, these data support that an early and pronounced activation of caspases can drive cells to undergo a form of necrotic-like regulated cell death.

  6. How much structuring is beneficial with regard to examination scores? A prospective study of three forms of active learning.

    PubMed

    Reinhardt, Claus H; Rosen, Evelyne N

    2012-09-01

    Many studies have demonstrated a superiority of active learning forms compared with traditional lecture. However, there is still debate as to what degree structuring is necessary with regard to high exam outcomes. Seventy-five students from a premedical school were randomly attributed to an active lecture group, a cooperative group, or a collaborative learning group. The active lecture group received lectures with questions to resolve at the end of the lecture. At the same time, the cooperative group and the collaborative group had to work on a problem and prepare presentations for their answers. The collaborative group worked in a mostly self-directed manner; the cooperative group had to follow a time schedule. For the additional work of preparing the poster presentation, the collaborative and cooperative groups were allowed 50% more working time. In part 1, all groups worked on the citric acid cycle, and in part 2, all groups worked on molecular genetics. Collaborative groups had to work on tasks and prepare presentations for their answers. At the end of each part, all three groups were subjected to the same exam. Additionally, in the collaborative and cooperative groups, the presentations were marked. All evaluations were performed by two independent examiners. Exam results of the active lecture groups were highest. Results of the cooperative group were nonsignificantly lower than the active lecture group and significantly higher than the collaborative group. The presentation quality was nonsignificantly higher in the collaborative group compared with the cooperative group. This study shows that active lecturing produced the highest exam results, which significantly differed from collaborative learning results. The additional elaboration in the cooperative and collaborative learning setting yielded the high presentation quality but apparently could not contribute further to exam scores. Cooperative learning seems to be a good compromise if high exam and

  7. The General Form of the Gibbs-Duhem Equation for Multiphase/Multicomponent Systems and Its Application to Solid-State Activity Measurements

    NASA Astrophysics Data System (ADS)

    Sacchetti, Mark

    2001-02-01

    The general form of the Gibbs-Duhem equation is derived for multiphase/multicomponent thermodynamic systems. The result illustrates that the form of the equation is invariant with respect to the number of phases present in the material. Therefore, calculation of activities or activity coefficients is possible using the same form of equation as has been applied to single-phase systems. The general form of the equation is applied to two organic solids, crystalline albuterol sulfate and amorphous poly(vinyl pyrrolidone). Physical chemistry instructors may find the derivation of the general form of the Gibbs-Duhem equation and its application to pharmaceutical materials a useful exercise for students.

  8. Excess TNF-α in the blood activates monocytes with the potential to directly form cholesteryl ester-laden cells.

    PubMed

    Zhu, Ming; Lei, Lei; Zhu, Zhenhua; Li, Qin; Guo, Dongqing; Xu, Jiajia; Chen, Jia; Sha, Huifang; Zhang, Xiaowei; Yang, Xinying; Song, Baoliang; Li, Boliang; Yan, Yan; Xiong, Ying

    2015-11-01

    The tumor necrosis factor-α (TNF-α) and monocytic cells play a critical role in the development of atherosclerosis, which is the major cause of coronary heart disease (CHD). In this work, we investigated the effect of excess TNF-α on monocytes in the blood and found that blood monocytes from the CHD patients had the potential to directly form cholesteryl ester (CE)-laden cells under the in vitro incubation with oxLDL. The plasma levels of proinflammatory cytokines, such as TNF-α, interleukin 6 (IL-6), and C reactive protein (CRP), in the CHD patients were significantly higher than those in the control healthy volunteers. However, only the plasma level of TNF-α, but not of IL-6 or CRP, is positively correlated with the potential of blood monocytes to directly form CE-laden cells. By using human blood monocytes and monocytic THP-1 cells, the activating effect of TNF-α on the formation of the CE-laden cells was demonstrated, which could be specifically blocked by the anti-TNF-α antibody. Furthermore, it was also revealed that TNF-α could boost adhesion and oxLDL uptake of the monocytes by enhancing the expression of the functional adhesion molecules and scavenger receptors, respectively. Finally, the results of in vivo and in vitro experiments with a mouse model confirmed that excess TNF-α in the blood activates monocytes with the potential to directly form CE-laden cells. These data demonstrate that excess TNF-α in the blood is the primary trigger for the development of atherosclerosis and CHD.

  9. Isolation, characterization and structure-elicitor activity relationships of hibernalin and its two oxidized forms from Phytophthora hibernalis Carne 1925.

    PubMed

    Capasso, Renato; Di Maro, Antimo; Cristinzio, Gennaro; De Martino, Antonio; Chambery, Angela; Daniele, Addolorata; Sannino, Filomena; Testa, Antonino; Parente, Augusto

    2008-01-01

    Three alpha-elicitins, named hibernalin1, hibernalin2 and hibernalin3 (hib1, hib2 and hib3, respectively), were isolated by reverse phase-low-pressure liquid chromatography from culture filtrates of Phytophthora hibernalis Carne 1925, the causal agent of citrus lemon brown rot. Hib1 proved to be identical to syringicin previously isolated from culture filtrates of Phytophthora syringae. Hib2 and hib3 shared the same primary structure with hib1, but contained, at position 50, Met sulphoxide or sulphone, respectively. By SDS-PAGE, the three proteins showed the same electrophoretic mobility, corresponding to about 10 kDa. Exact M(r) values were obtained by MALDI-TOF-MS (10,194.82 for hib1, 10,209.33 for hib2 and 10,223.80 for hib3), while by ESI-MS an M(r) value of 10,194.90 was found for hib1 and no results for hib2 and hib3. The hibernalin forms showed a high propensity to self-association, after exposure to acetonitrile. Hib1 showed to be active in both the hypersensitivity response and electrolytes leakage assays; the sample containing hib1 and hib2 was only weakly active in the first assay and inactive in the second assay, while the sample containing all three hibernalin forms proved to be inactive in both tests. It is proposed that the different activities of the three hibernalin samples could be very likely attributed to both Met50 oxidation and aggregation.

  10. The role of highly purified forms of rat liver cytochrome P-450 in the dimethylation of dimethylnitrosamine and its activation to mutagens.

    PubMed

    Masson, H A; Ioannides, C; Gibson, G G

    1983-06-01

    Highly purified NADPH-cytochrome P-450 reductase and the major phenobarbital (PB) and beta-naphthoflavone (beta NF) forms of cytochrome P-450 were used in reconstituted systems to study the demethylation and subsequent activation of dimethylnitrosamine (DMN) to mutagenic intermediates. Both forms of cytochrome P-450 were active in the demethylation of DMN, cytochrome P-450 from PB-treated animals being more efficient, generating nearly twice as much formaldehyde per nmol of haemoprotein. Neither form of the cytochrome could activate DMN to mutagens in the Ames test. These findings indicate that DMN demethylation does not lead to its activation to mutagenic products.

  11. Substrate-Mediated Oxygen Activation by Homoprotocatechuate 2,3-Dioxygenase: Intermediates Formed by a Tyrosine 257 Variant

    PubMed Central

    Mbughuni, Michael M.; Meier, Katlyn K.; Münck, Eckard; Lipscomb, John D.

    2012-01-01

    Homoprotocatechuate (HPCA; 3,4-dihydroxyphenylacetate or 4-carboxymethyl catechol) and O2 bind in adjacent ligand sites of the active site FeII of Homoprotocatechuate 2,3-Dioxygenase (FeHPCD). We have proposed that electron transfer from the chelated aromatic substrate through the FeII to O2 gives both substrates radical character. This would promote reaction between the substrates to form an alkylperoxo intermediate as the first step in aromatic ring cleavage. Several active site amino acids are thought to promote these reactions through acid/base chemistry, hydrogen bonding, and electrostatic interactions. Here the role of Tyr257 is explored by using the Tyr257Phe (Y257F) variant, which decreases kcat by about 75%. The crystal structure of the FeHPCD-HPCA complex has shown that Tyr257 hydrogen bonds to the deprotonated C2-hydroxyl of HPCA. Stopped-flow studies show that at least two reaction intermediates, termed Y257FInt1HPCA and Y257FInt2HPCA, accumulate during the Y257F-HPCA + O2 reaction prior to formation of the ring-cleaved product. Y257FInt1HPCA is colorless and is formed as O2 binds reversibly to the HPCA-enzyme complex. Y257FInt2HPCA forms spontaneously from Y257FInt1HPCA and displays a chromophore at 425 nm (ε425 = 10,500 M-1 cm−1). Mössbauer spectra of the intermediates trapped by rapid freeze quench show that both intermediates contain FeII. The lack of a chromophore characteristic of a quinone or semiquinone form of HPCA, the presence of FeII, and the low O2 affinity suggests that Y257FInt1HPCA is an HPCA-FeII-O2 complex with little electron delocalization onto the O2. In contrast, the intense spectrum of Y257FInt2HPCA suggests the intermediate is most likely an HPCA quinone-FeII-(hydro)peroxo species. Steady-state and transient kinetic analyses show that steps of the catalytic cycle are slowed by as much as 100-fold by the mutation. These effects can be rationalized by a failure of Y257F to facilitate the observed distortion of the bound HPCA

  12. Comparison of the Physical Activity and Sedentary Behaviour Assessment Questionnaire and the Short-Form International Physical Activity Questionnaire: An Analysis of Health Survey for England Data

    PubMed Central

    Scholes, Shaun; Bridges, Sally; Ng Fat, Linda; Mindell, Jennifer S.

    2016-01-01

    Background The Physical Activity and Sedentary Behaviour Assessment Questionnaire (PASBAQ), used within the Health Survey for England (HSE) at 5-yearly intervals, is not included annually due to funding and interview-length constraints. Policy-makers and data-users are keen to consider shorter instruments such as the Short-form International Physical Activity Questionnaire (IPAQ) for the annual survey. Both questionnaires were administered in HSE 2012, enabling comparative assessment in a random sample of 1252 adults. Methods Relative agreement using prevalence-adjusted bias-adjusted Kappa (PABAK) statistics was estimated for: sufficient aerobic activity (moderate-to-vigorous physical activity [MVPA] ≥150minutes/week); inactivity (MVPA<30minutes/week); and excessive sitting (≥540minutes/weekday). Cross-sectional associations with health outcomes were compared across tertiles of MVPA and tertiles of sitting time using logistic regression with tests for linear trend. Results Compared with PASBAQ data, IPAQ-assessed estimates of sufficient aerobic activity and inactivity were higher and lower, respectively; estimates of excessive sitting were higher. Demographic patterns in prevalence were similar. Agreement using PABAK statistics was fair-to-moderate for sufficient aerobic activity (0.32–0.49), moderate-to-substantial for inactivity (0.42–0.74), and moderate-to-substantial for excessive sitting (0.49–0.75). As with the PASBAQ, IPAQ-assessed MVPA and sitting each showed graded associations with mental well-being (women: P for trend = 0.003 and 0.004, respectively) and obesity (women: P for trend = 0.007 and 0.014, respectively). Conclusions Capturing habitual physical activity and sedentary behaviour through brief questionnaires is complex. Differences in prevalence estimates can reflect differences in questionnaire structure and content rather than differences in reported behaviour. Treating all IPAQ-assessed walking as moderate-intensity contributed to the

  13. Evaluation of in vivo antimycobacterial activity of some folklore medicinal plants and enumeration of colony forming unit in murine model

    PubMed Central

    Barua, Acheenta Gohain; Raj, Himangshu; Konch, Pranab; Hussain, P.; Barua, Chandana C.

    2016-01-01

    Objectives: The present study was carried out to investigate the in vivo antimycobacterial activity of methanol extract of Alstonia scholaris and Mucuna imbricata in murine model. Materials and Methods: Female BALB/c mice were infected with the Mycobacterium tuberculosis H37Rv suspension. Extracts were administered orally for 2 weeks from 7th day postinfection at a dose of 200 mg/kg and rifampicin at 20 mg/kg as standard. The synergistic groups were 10 and 100 mg/kg for rifampicin and extract, respectively. Results: The final body weight of mycobacteria-infected group was significantly reduced (15.41 ± 0.42, P < 0.01), but following treatment with the plant extract plus rifampicin could elevate the body weight. Colony forming unit (CFU) count of lung (8.71 ± 0.01) and spleen (8.59 ± 0.01) was significantly higher in infected and untreated group (P < 0.01). It was observed that activity of the synergistic group displayed powerful and maximum response against tuberculosis (TB) infection with lower CFU counts. Histopathology study showed cells such as lymphocytes, epithelioid, Langhans giant cell, and fibrous tissue proliferation in lungs; depletion of lymphocytes in the spleen. Conclusions: The data indicate that methanol extract of A. scholaris has potential antimycobacterial activity, and the synergistic group consisting of rifampicin and A. scholaris could be a rational choice for the treatment of TB. PMID:27721538

  14. Proteolytic cleavage of chemerin protein is necessary for activation to the active form, Chem157S, which functions as a signaling molecule in glioblastoma.

    PubMed

    Yamaguchi, Yasuto; Du, Xiao-Yan; Zhao, Lei; Morser, John; Leung, Lawrence L K

    2011-11-11

    Chemerin is a chemoattractant involved in innate and adaptive immunity as well as an adipokine implicated in adipocyte differentiation. Chemerin circulates as an inactive precursor in blood whose bioactivity is closely regulated through proteolytic processing at its C terminus. We developed methodology for production of different recombinant chemerin isoforms (chem163S, chem157S, and chem155A) which allowed us to obtain large quantities of these proteins with purity of >95%. Chem158K was generated from chem163S by plasmin cleavage. Characterization by mass spectrometry and Edman degradation demonstrated that both the N and C termini were correct for each isoform. Ca(2+) mobilization assays showed that the EC(50) values for chem163S and chem158K were 54.2 ± 19.9 nm and 65.2 ± 13.2 nm, respectively, whereas chem157S had a ∼50-fold higher potency with an EC(50) of 1.2 ± 0.7 nm. Chem155A had no agonist activity and weak antagonist activity, causing a 50% reduction of chem157S activity at a molar ratio of 100:1. Similar results were obtained in a chemotaxis assay. Because chem158K is the dominant form in cerebrospinal fluid from patients with glioblastoma (GBM), we examined the significance of chemerin in GBM biology. In silico analysis showed chemerin mRNA was significantly increased in tissue from grade III and IV gliomas. Furthermore, U-87 MG cells, a human GBM line, express the chemerin receptors, chemokine-like receptor 1 and chemokine receptor-like 2, and chem157S triggered Ca(2+) flux. This study emphasized the necessity of appropriate C-terminal proteolytic processing to generate the likely physiologic form of active chemerin, chem157S, and suggested a possible role in malignant GBM.

  15. Importance of clustered 2'-O-(2-aminoethyl) residues for the gene targeting activity of triple helix-forming oligonucleotides.

    PubMed

    Puri, Nitin; Majumdar, Alokes; Cuenoud, Bernard; Miller, Paul S; Seidman, Michael M

    2004-02-10

    We are developing triple helix-forming oligonucleotides (TFOs) as gene targeting reagents in living mammalian cells. We have described psoralen-linked TFOs with 2'-O-methyl and 2'-O-(2-aminoethyl) (2'-AE) substitutions that are active in a gene knockout assay in cultured cells. The assay is based on mutagenesis by psoralen, a photoactive DNA cross-linker. Previous work showed that TFOs with three or four 2'-AE residues were disproportionately more active than those with one or two substitutions. Here we demonstrate that for optimal bioactivity the 2'-AE residues must be clustered rather than dispersed. We have further characterized bioactive and inactive TFOs in an effort to identify biochemical and biophysical correlates of biological activity. While thermal stability is a standard monitor of TFO biophysical activity, we find that T(m) values do not distinguish bioactive and inactive TFOs. In contrast, measurements of TFO association rates appear to correlate well with bioactivity, in that triplex formation occurs disproportionately faster with the TFOs containing three or four 2'-AE residues. We asked if extending the incubation time prior to photoactivation would enhance the bioactivity of a TFO with a slow on rate relative to the TFO with a faster association rate. However, there was no change in bioactivity differential. These results are compatible with a model in which TFO binding in vivo is followed by relatively rapid elution by cellular functions, similar to that described for transcription factors. Under these circumstances, TFOs with faster on rates would be favored because they would be more likely to be in triplexes at the time of photoactivation.

  16. Channel-forming activity of syringopeptin 25A in mercury-supported lipid bilayers with a phosphatidylcholine distal leaflet.

    PubMed

    Becucci, Lucia; Rossi, Marta; Fiore, Alberto; Scaloni, Andrea; Guidelli, Rolando

    2016-04-01

    The channel-forming activity of the lipodepsipeptide syringopeptin 25A (SP25A) was investigated at a tethered bilayer lipid membrane (tBLM) with a dioleoylphosphatidylcholine distal leaflet, anchored to a mercury electrode through a hydrophilic tetraethyleneoxy spacer. SP25A was incorporated in the tBLM from different aqueous solutions by recording a series of impedance spectra over a potential range encompassing non-physiological transmembrane potential (Δϕ) values. Once incorporated, SP25A forms stable ion channels over the narrower range of physiological Δϕ values. Ion flow into and out of the spacer, through the lipid bilayer moiety of the tBLM, was monitored by potential step chronocoulometry and cyclic voltammetry at pH3, 5.4 and 6.8. Potassium ion flow into the hydrophilic spacer along the SP25A channels, during the negative potential scan, proceeds in two stages, except at the higher pH and lower SP25A concentration adopted, where it proceeds in a single stage. In light of the behavior of SP25A single channel currents reported in the literature, the first stage is ascribed to large channels resulting from the aggregation of small ones, while the second more negative stage is associated with the small channels resulting from the disaggregation of the large ones.

  17. Novel Synthesis Method of Micronized Ti-Zeolite Na-A and Cytotoxic Activity of Its Silver Exchanged Form

    PubMed Central

    Youssef, H. F.; Hegazy, W. H.; Abo-almaged, H. H.; El-Bassyouni, G. T.

    2015-01-01

    The core-shell method is used as a novel synthetic process of micronized Ti-Zeolite Na-A which involves calcination at 700°C of coated Egyptian Kaolin with titanium tetrachloride in acidic medium as the first step. The produced Ti-coated metakaolinite is subjected to microwave irradiation at low temperature of 80°C for 2 h. The prepared micronized Ti-containing Zeolites-A (Ti-Z-A) is characterized by FTIR, XRF, XRD, SEM, and EDS elemental analysis. Ag-exchanged form of Ti-Z-Ag is also prepared and characterized. The Wt% of silver exchanged onto the Ti-Zeolite structure was determined by atomic absorption spectra. The in vitro cytotoxic activity of Ti-Z-Ag against human hepatocellular carcinoma cell line (HePG2), colon cell line carcinoma (HCT116), lung carcinoma cell line (A549), and human Caucasian breast adenocarcinoma (MCF7) is reported. The results were promising and revealed that the exchanged Ag form of micronized Ti-Zeolite-A can be used as novel antitumor drug. PMID:25705142

  18. Disrupting a key hydrophobic pair in the oligomerization interface of the actinoporins impairs their pore-forming activity.

    PubMed

    Mesa-Galloso, Haydeé; Delgado-Magnero, Karelia H; Cabezas, Sheila; López-Castilla, Aracelys; Hernández-González, Jorge E; Pedrera, Lohans; Alvarez, Carlos; Peter Tieleman, D; García-Sáez, Ana J; Lanio, Maria E; Ros, Uris; Valiente, Pedro A

    2017-03-01

    Crystallographic data of the dimeric and octameric forms of fragaceatoxin C (FraC) suggested the key role of a small hydrophobic protein-protein interaction surface for actinoporins oligomerization and pore formation in membranes. However, site-directed mutagenesis studies supporting this hypothesis for others actinoporins are still lacking. Here, we demonstrate that disrupting the key hydrophobic interaction between V60 and F163 (FraC numbering scheme) in the oligomerization interface of FraC, equinatoxin II (EqtII), and sticholysin II (StII) impairs the pore formation activity of these proteins. Our results allow for the extension of the importance of FraC protein-protein interactions in the stabilization of the oligomeric intermediates of StII and EqtII pointing out that all of these proteins follow a similar pathway of membrane disruption. These findings support the hybrid pore proposal as the universal model of actinoporins pore formation. Moreover, we reinforce the relevance of dimer formation, which appears to be a functional intermediate in the assembly pathway of some different pore-forming proteins.

  19. Ubiquitin-dependent proteolytic pathway in wheat germ: Isolation of multiple forms of ubiquitin-activating enzyme, E1

    SciTech Connect

    Hatfield, P.M.; Vierstra, R.D. )

    1989-01-24

    Ubiquitin is a highly conserved protein involved in several important regulatory processes through its ATP-dependent, covalent ligation to a variety of eukaryotic target proteins. The authors describe here the characterization of ubiquitin conjugation in wheat germ extracts and the subsequent isolation of enzymes involved in conjugation. With {sup 125}I-ubiquitin as a substrate, wheat germ extracts form conjugates with either endogenous or added proteins. Ubiquitin-activating enzyme (E1) was purified from wheat germ extracts by using a modification of the covalent affinity chromatography procedure of Ciechanover et al. E1 from wheat germ, like that from rabbit reticulocytes, formed thiol ester intermediates with ubiquitin in the presence of ATP. Purified E1 preparations contained three polypeptides of apparent molecular masses of 117, 123, and 126 kDa after NaDodSO{sub 4}-PAGE. Under nondenaturing conditions, these proteins have native molecular masses of {approx}115 kDa, indicating that they exist as monomers. They concluded that all three species were E1 on the basis of their coelution with E1 activity, by immunorecognition by anti-human E1 antibodies, and by the similarity of their peptide maps. Furthermore, antibodies prepared against wheat germ E1's recognized E1 from rabbit reticulocytes. All three wheat germ E1's were detected in crude extracts prepared under conditions that minimized proteolysis, suggesting that the heterogeneity of the purified E1 preparations was not the result of posthomogenization breakdown. The immunological similarity of animal and plant E1's indicates that this conjugation enzyme, like ubiquitin, has been conserved through evolution.

  20. Active optics and modified-Rumsey wide-field telescopes: MINITRUST demonstrators with vase- and tulip-form mirrors

    NASA Astrophysics Data System (ADS)

    Lemaître, Gérard R.; Montiel, Pierre; Joulié, Patrice; Dohlen, Kjetil; Lanzoni, Patrick

    2005-12-01

    Wide-field astronomy requires the development of larger aperture telescopes. The optical properties of a three-mirror modified-Rumsey design provide significant advantages when compared to other telescope designs: (i) at any wavelength, the design has a flat field and is anastigmatic; (ii) the system is extremely compact, i.e., it is almost four times shorter than a Schmidt. Compared to the equally compact flat-field Ritchey-Chrétien with a doublet-lens corrector, as developed for the Sloan digital sky survey - and which requires the polishing of six optical surfaces - the proposed modified-Rumsey design requires only a two-surface polishing and provides a better imaging quality. All the mirrors are spheroids of the hyperboloid type. Starting from the classical Rumsey design, it is shown that the use of all eight available free parameters allows the simultaneous aspherization of the primary and tertiary mirrors by active optics methods from a single deformable substrate. The continuity conditions between the primary and the tertiary hyperbolizations are achieved by an intermediate narrow ring of constant thickness that is not optically used. After the polishing of a double vase form in a spherical shape, the primary-tertiary hyperbolizations are achieved by in situ stressing. The tulip-form secondary is hyperbolized by stress polishing. Other active optics alternatives are possible for a space telescope. The modified-Rumsey design is of interest for developing large space- and ground-based survey telescopes in UV, visible, or IR ranges, such as currently demonstrated with the construction of identical telescopes MINITRUST-1 and -2, f/5 - 2° field of view. Double-pass optical tests show diffraction-limited images.

  1. Active optics and modified-Rumsey wide-field telescopes: MINITRUST demonstrators with vase- and tulip-form mirrors.

    PubMed

    Lemaître, Gérard R; Montiel, Pierre; Joulié, Patrice; Dohlen, Kjetil; Lanzoni, Patrick

    2005-12-01

    Wide-field astronomy requires the development of larger aperture telescopes. The optical properties of a three-mirror modified-Rumsey design provide significant advantages when compared to other telescope designs: (i) at any wavelength, the design has a flat field and is anastigmatic; (ii) the system is extremely compact, i.e., it is almost four times shorter than a Schmidt. Compared to the equally compact flat-field Ritchey-Chrétien with a doublet-lens corrector, as developed for the Sloan digital sky survey-and which requires the polishing of six optical surfaces-the proposed modified-Rumsey design requires only a two-surface polishing and provides a better imaging quality. All the mirrors are spheroids of the hyperboloid type. Starting from the classical Rumsey design, it is shown that the use of all eight available free parameters allows the simultaneous aspherization of the primary and tertiary mirrors by active optics methods from a single deformable substrate. The continuity conditions between the primary and the tertiary hyperbolizations are achieved by an intermediate narrow ring of constant thickness that is not optically used. After the polishing of a double vase form in a spherical shape, the primary-tertiary hyperbolizations are achieved by in situ stressing. The tulip-form secondary is hyperbolized by stress polishing. Other active optics alternatives are possible for a space telescope. The modified-Rumsey design is of interest for developing large space- and ground-based survey telescopes in UV, visible, or IR ranges, such as currently demonstrated with the construction of identical telescopes MINITRUST-1 and -2, f/5-2 degrees field of view. Double-pass optical tests show diffraction-limited images.

  2. The closed/open model for lipase activation. Addressing intermediate active forms of fungal enzymes by trapping of conformers in water-restricted environments.

    PubMed

    González-Navarro, H; Bañó, M C; Abad, C

    2001-03-13

    The behavior of prototypic fungal lipases in a water-restricted environment has been investigated by exploiting the reported experimental strategy that allows the trapping (freeze-drying) of the enzyme in the conformation present in aqueous solution and to subsequently assay it in nonaqueous media [Mingarro, I., Abad, C., and Braco, L. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 3308-3312]. We now report, using simple esterification as well as acidolysis (triglycerides as substrates) as nonaqueous model reactions, that the presence of a detergent (n-octyl-beta-glucopyranoside) in the freeze-drying buffer, at concentrations below the critical micellar concentration, generates different catalytically active (kinetically trapped) conformational states of the enzyme. These activated forms exquisitely discriminate between short- and long-chain fatty acids, suggesting that they can be correlated with intermediate conformations of the protein sufficiently open to permit the access of relatively small but not large substrates. Additional data obtained from aqueous solution activity measurements in the presence of detergent revealed that the fungal lipase retains an active conformation induced by high detergent concentration (30 mM) for a long period of time, a 'memory effect', which is stabilized in the absence of a well-defined interface by few detergent molecules. Together these results provide support to a model of lipase action involving several equilibrium states (closed, intermediate, and open), which can be modulated by the composition of the microenvironment, i.e., by the detergent concentration.

  3. The mRNA-edited form of GABRA3 suppresses GABRA3-mediated Akt activation and breast cancer metastasis.

    PubMed

    Gumireddy, Kiranmai; Li, Anping; Kossenkov, Andrew V; Sakurai, Masayuki; Yan, Jinchun; Li, Yan; Xu, Hua; Wang, Jian; Zhang, Paul J; Zhang, Lin; Showe, Louise C; Nishikura, Kazuko; Huang, Qihong

    2016-02-12

    Metastasis is a critical event affecting breast cancer patient survival. To identify molecules contributing to the metastatic process, we analysed The Cancer Genome Atlas (TCGA) breast cancer data and identified 41 genes whose expression is inversely correlated with survival. Here we show that GABAA receptor alpha3 (Gabra3), normally exclusively expressed in adult brain, is also expressed in breast cancer, with high expression of Gabra3 being inversely correlated with breast cancer survival. We demonstrate that Gabra3 activates the AKT pathway to promote breast cancer cell migration, invasion and metastasis. Importantly, we find an A-to-I RNA-edited form of Gabra3 only in non-invasive breast cancers and show that edited Gabra3 suppresses breast cancer cell invasion and metastasis. A-to-I-edited Gabra3 has reduced cell surface expression and suppresses the activation of AKT required for cell migration and invasion. Our study demonstrates a significant role for mRNA-edited Gabra3 in breast cancer metastasis.

  4. Enhanced electrochemical performance of porous activated carbon by forming composite with graphene as high-performance supercapacitor electrode material

    NASA Astrophysics Data System (ADS)

    Wang, Zhi-Hang; Yang, Jia-Ying; Wu, Xiong-Wei; Chen, Xiao-Qing; Yu, Jin-Gang; Wu, Yu-Ping

    2017-02-01

    In this work, a novel activated carbon containing graphene composite was developed using a fast, simple, and green ultrasonic-assisted method. Graphene is more likely a framework which provides support for activated carbon (AC) particles to form hierarchical microstructure of carbon composite. Scanning electron microscope (SEM), transmission electron microscope (TEM), Brunauer-Emmett-Teller (BET) surface area measurement, thermogravimetric analysis (TGA), Raman spectra analysis, XRD, and XPS were used to analyze the morphology and surface structure of the composite. The electrochemical properties of the supercapacitor electrode based on the as-prepared carbon composite were investigated by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), charge/discharge, and cycling performance measurements. It exhibited better electrochemical performance including higher specific capacitance (284 F g-1 at a current density of 0.5 A g-1), better rate behavior (70.7% retention), and more stable cycling performance (no capacitance fading even after 2000 cycles). It is easier for us to find that the composite produced by our method was superior to pristine AC in terms of electrochemical performance due to the unique conductive network between graphene and AC.

  5. Active methylotrophs in the sediments of Lonar Lake, a saline and alkaline ecosystem formed by meteor impact.

    PubMed

    Antony, Chakkiath Paul; Kumaresan, Deepak; Ferrando, Lucia; Boden, Rich; Moussard, Hélène; Scavino, Ana Fernández; Shouche, Yogesh S; Murrell, J Colin

    2010-11-01

    Lonar Lake is a unique saline and alkaline ecosystem formed by meteor impact in the Deccan basalts in India around 52,000 years ago. To investigate the role of methylotrophy in the cycling of carbon in this unusual environment, stable-isotope probing (SIP) was carried out using the one-carbon compounds methane, methanol and methylamine. Denaturing gradient gel electrophoresis fingerprinting analyses performed with heavy (13)C-labelled DNA retrieved from sediment microcosms confirmed the enrichment and labelling of active methylotrophic communities. Clone libraries were constructed using PCR primers targeting 16S rRNA genes and functional genes. Methylomicrobium, Methylophaga and Bacillus spp. were identified as the predominant active methylotrophs in methane, methanol and methylamine SIP microcosms, respectively. Absence of mauA gene amplification in the methylamine SIP heavy fraction also indicated that methylamine metabolism in Lonar Lake sediments may not be mediated by the methylamine dehydrogenase enzyme pathway. Many gene sequences retrieved in this study were not affiliated with extant methanotrophs or methylotrophs. These sequences may represent hitherto uncharacterized novel methylotrophs or heterotrophic organisms that may have been cross-feeding on methylotrophic metabolites or biomass. This study represents an essential first step towards understanding the relevance of methylotrophy in the soda lake sediments of an unusual impact crater structure.

  6. The mRNA-edited form of GABRA3 suppresses GABRA3-mediated Akt activation and breast cancer metastasis

    PubMed Central

    Gumireddy, Kiranmai; Li, Anping; Kossenkov, Andrew V.; Sakurai, Masayuki; Yan, Jinchun; Li, Yan; Xu, Hua; Wang, Jian; Zhang, Paul J.; Zhang, Lin; Showe, Louise C.; Nishikura, Kazuko; Huang, Qihong

    2016-01-01

    Metastasis is a critical event affecting breast cancer patient survival. To identify molecules contributing to the metastatic process, we analysed The Cancer Genome Atlas (TCGA) breast cancer data and identified 41 genes whose expression is inversely correlated with survival. Here we show that GABAA receptor alpha3 (Gabra3), normally exclusively expressed in adult brain, is also expressed in breast cancer, with high expression of Gabra3 being inversely correlated with breast cancer survival. We demonstrate that Gabra3 activates the AKT pathway to promote breast cancer cell migration, invasion and metastasis. Importantly, we find an A-to-I RNA-edited form of Gabra3 only in non-invasive breast cancers and show that edited Gabra3 suppresses breast cancer cell invasion and metastasis. A-to-I-edited Gabra3 has reduced cell surface expression and suppresses the activation of AKT required for cell migration and invasion. Our study demonstrates a significant role for mRNA-edited Gabra3 in breast cancer metastasis. PMID:26869349

  7. The importance of the activation volume for the description of the molecular dynamics of glass-forming liquids

    NASA Astrophysics Data System (ADS)

    Kaminski, K.; Pawlus, S.; Adrjanowicz, K.; Wojnarowska, Z.; Wlodarczyk, P.; Paluch, M.

    2012-02-01

    High pressure dielectric measurements were carried out on hydrogen bonded d-glucose and two van der Waals peracetyl saccharides, i.e. α pentaacetyl glucose and α octaacetyl maltose. In this study we found that after removing H bonds, the molecular dynamics of both modified saccharides is very sensitive to pressure, as reflected by a large value of the pressure coefficient of the glass transition temperature, equal to 270 K GPa-1 and 280 K GPa-1 for α pentaacetyl glucose and α octaacetyl maltose, respectively. On the other hand, dTg/dP for d-glucose is much lower, equal to 67 K GPa-1. Our result confirms the general rule that the hydrogen bonding glass-forming liquids exhibit much lower values of dTg/dP compared to the van der Waals systems. Additionally, on the basis of results reported herein and also recent literature data for polyalcohols, we point out that the activation volume correlates fairly well with the molecular volume in the case of hydrogen bonding liquids. On the other hand, much larger values of the activation volumes at Tg with respect to the molecular volumes were found for both peracetyl saccharides.

  8. A novel form of synaptic plasticity in field CA3 of hippocampus requires GPER1 activation and BDNF release.

    PubMed

    Briz, Victor; Liu, Yan; Zhu, Guoqi; Bi, Xiaoning; Baudry, Michel

    2015-09-28

    Estrogen is an important modulator of hippocampal synaptic plasticity and memory consolidation through its rapid action on membrane-associated receptors. Here, we found that both estradiol and the G-protein-coupled estrogen receptor 1 (GPER1) specific agonist G1 rapidly induce brain-derived neurotrophic factor (BDNF) release, leading to transient stimulation of activity-regulated cytoskeleton-associated (Arc) protein translation and GluA1-containing AMPA receptor internalization in field CA3 of hippocampus. We also show that type-I metabotropic glutamate receptor (mGluR) activation does not induce Arc translation nor long-term depression (LTD) at the mossy fiber pathway, as opposed to its effects in CA1, and it only triggers LTD after GPER1 stimulation. Furthermore, this form of mGluR-dependent LTD is associated with ubiquitination and proteasome-mediated degradation of GluA1, and is prevented by proteasome inhibition. Overall, our study identifies a novel mechanism by which estrogen and BDNF regulate hippocampal synaptic plasticity in the adult brain.

  9. High-level expression and purification of human xylosyltransferase I in High Five insect cells as biochemically active form.

    PubMed

    Kuhn, Joachim; Müller, Sandra; Schnölzer, Martina; Kempf, Tore; Schön, Sylvia; Brinkmann, Thomas; Schöttler, Manuela; Götting, Christian; Kleesiek, Knut

    2003-12-19

    Human xylosyltransferase I (XT-I) catalyzes the transfer of xylose from UDP-xylose to consensus serine residues of proteoglycan core proteins. Expression of a soluble form of recombinant histidine-tagged XT-I (rXT-I-HIS) was accomplished at a high level with High Five/pCG255-1 insect cells in suspension culture. The recombinant protein was purified to homogeneity by a combination of heparin affinity chromatography and metal (Ni(2+)) chelate affinity chromatography. Using the modern technique of perfusion chromatography, a rapid procedure for purification of the rXT-I-HIS from insect cell culture supernatant was developed. The purified, biologically active enzyme was homogeneous on SDS-PAGE, was detected with anti-XT-I-antibodies, and had the expected tryptic fragment mass spectrum. N-terminal amino acid sequencing demonstrated that the N-terminal signal sequence of the expressed protein was quantitatively cleaved. The total yield of the enzyme after purification was 18% and resulted in a specific XT-I activity of 7.9mU/mg. The K(m) of the enzyme for recombinant [Val(36),Val(38)](delta1),[Gly(92),Ile(94)](delta2)bikunin was 0.8microM. About 5mg purified enzyme could be obtained from 1L cell culture supernatant. The availability of substantial quantities of active, homogeneous enzyme will be of help in future biochemical and biophysical characterization of XT-I and for the development of a immunological XT-I assay.

  10. Neural progenitor cells isolated from the subventricular zone present hemichannel activity and form functional gap junctions with glial cells

    PubMed Central

    Talaverón, Rocío; Fernández, Paola; Escamilla, Rosalba; Pastor, Angel M.; Matarredona, Esperanza R.; Sáez, Juan C.

    2015-01-01

    The postnatal subventricular zone (SVZ) lining the walls of the lateral ventricles contains neural progenitor cells (NPCs) that generate new olfactory bulb interneurons. Communication via gap junctions between cells in the SVZ is involved in NPC proliferation and in neuroblast migration towards the olfactory bulb. SVZ NPCs can be expanded in vitro in the form of neurospheres that can be used for transplantation purposes after brain injury. We have previously reported that neurosphere-derived NPCs form heterocellular gap junctions with host glial cells when they are implanted after mechanical injury. To analyze functionality of NPC-glial cell gap junctions we performed dye coupling experiments in co-cultures of SVZ NPCs with astrocytes or microglia. Neurosphere-derived cells expressed mRNA for at least the hemichannel/gap junction channel proteins connexin 26 (Cx26), Cx43, Cx45 and pannexin 1 (Panx1). Dye coupling experiments revealed that gap junctional communication occurred among neurosphere cells (incidence of coupling: 100%). Moreover, hemichannel activity was also detected in neurosphere cells as evaluated in time-lapse measurements of ethidium bromide uptake. Heterocellular coupling between NPCs and glial cells was evidenced in co-cultures of neurospheres with astrocytes (incidence of coupling: 91.0 ± 4.7%) or with microglia (incidence of coupling: 71.9 ± 6.7%). Dye coupling in neurospheres and in co-cultures was inhibited by octanol, a gap junction blocker. Altogether, these results suggest the existence of functional hemichannels and gap junction channels in postnatal SVZ neurospheres. In addition, they demonstrate that SVZ-derived NPCs can establish functional gap junctions with astrocytes or microglia. Therefore, cell-cell communication via gap junctions and hemichannels with host glial cells might subserve a role in the functional integration of NPCs after implantation in the damaged brain. PMID:26528139

  11. Characterization of the clotting activities of structurally different forms of activated factor IX. Enzymatic properties of normal human factor IXa alpha, factor IXa beta, and activated factor IX Chapel Hill.

    PubMed Central

    Griffith, M J; Breitkreutz, L; Trapp, H; Briet, E; Noyes, C M; Lundblad, R L; Roberts, H R

    1985-01-01

    Two structurally different forms of activated human Factor IX (Factor IXa alpha and IXa beta) have been previously reported to have essentially identical clotting activity in vitro. Although it has been shown that activated Factor IX Chapel Hill, an abnormal Factor IX isolated from the plasma of a patient with mild hemophilia B, and normal Factor IXa alpha are structurally very similar, the clotting activity of activated Factor IX Chapel Hill is much lower (approximately fivefold) than that of normal Factor IXa beta. In the present study we have prepared activated Factor IX by incubating human Factor IX with calcium and Russell's viper venom covalently bound to agarose. Fractionation of the activated Factor IX by high-performance liquid chromatography demonstrated the presence of both Factors IXa alpha and IXa beta. On the basis of active site concentration, determined by titration with antithrombin III, the clotting activities of activated Factor IX Chapel Hill and IXa alpha were similar, but both activities were less than 20% of the clotting activity of Factor IXa beta. Activated Factor IX activity was also measured in the absence of calcium, phospholipid, and Factor VIII, by determination of the rate of Factor X activation in the presence of polylysine. In the presence of polylysine, the rates of Factor X activation by activated Factor IX Chapel Hill, Factor IXa alpha, and Factor IXa beta were essentially identical. We conclude that the clotting activity of activated Factor IX Chapel Hill is reduced when compared with that of Factor IXa beta but essentially normal when compared with that of Factor IXa alpha. PMID:3871202

  12. Prognostic Significance of Active and Modified forms of Endothelin 1 in Patients with Heart Failure with Reduced Ejection Fraction

    PubMed Central

    Gottlieb, Stephen S.; Harris, Kristie; Todd, John; Estis, Joel; Christenson, Robert H.; Torres, Victoria; Whittaker, Kerry; Rebuck, Heather; Wawrzyniak, Andrew; Krantz, David S.

    2015-01-01

    Objectives Concentrations of endothelin I (ET1) are elevated in CHF patients, and, like other biomarkers that reflect hemodynamic status and cardiac pathophysiology, are prognostic. The Singulex assay (Sgx-ET1) measures the active form of ET1, with a short in-vivo half-life and C-terminal endothelin-1 (CT-ET1) is measured by the Brahms assay and is a modified (degraded) product with longer half-life. We aimed to determine the prognostic importance of active and modified forms of endothelin 1 (Singulex and Brahms assays) in comparison with other commonly measured biomarkers of inflammation, hemodynamic status and cardiac physiology in CHF. Design & Methods Plasma biomarkers (Sgx-ET1, CT-ET1, NTproBNP, IL-6, TNFα, cTnI, VEGF, hs-CRP, Galectin-3, ST2) were measured in 134 NYHA class II and III CHF patients with systolic dysfunction. Prognostic importance of biomarkers for hospitalization or death were calculated by both logistic regression and Kaplan-Meier survival analyses. Results CT-ET1 (OR 5.2, 95% CI 1.7–15.7) and Sgx-ET1 (OR 2.9, CI 1.1–7.7) were independent predictors of hospitalization and death and additively predicted events after adjusting for age, sex and other significant biomarkers. Other biomarkers did not improve the model. Similarly, in Cox regression analysis, only CT-ET1 (HR 3.4, 95% CI 1.4–8.4), VEGF (2.7, 95% CI 1.3–5.4) and Sgx-ET1 (HR 2.6, 95% CI 1.2–5.6) were independently prognostic. Conclusions Elevated concentrations of endothelin 1 predict mortality and hospitalizations in HF patients. Endothelin 1 was more prognostic than commonly obtained hemodynamic, inflammatory and fibrotic biomarkers. Two different assays of endothelin 1 independently and synergistically were prognostic, suggesting either complementary information or extreme prognostic importance. PMID:25541019

  13. Spectroscopic and crystallographic characterization of "alternative resting" and "resting oxidized" enzyme forms of bilirubin oxidase: implications for activity and electrochemical behavior of multicopper oxidases.

    PubMed

    Kjaergaard, Christian H; Durand, Fabien; Tasca, Federico; Qayyum, Munzarin F; Kauffmann, Brice; Gounel, Sébastien; Suraniti, Emmanuel; Hodgson, Keith O; Hedman, Britt; Mano, Nicolas; Solomon, Edward I

    2012-03-28

    While there is broad agreement on the catalytic mechanism of multicopper oxidases (MCOs), the geometric and electronic structures of the resting trinuclear Cu cluster have been variable, and their relevance to catalysis has been debated. Here, we present a spectroscopic characterization, complemented by crystallographic data, of two resting forms occurring in the same enzyme and define their interconversion. The resting oxidized form shows similar features to the resting form in Rhus vernicifera and Trametes versicolor laccase, characterized by "normal" type 2 Cu electron paramagnetic resonance (EPR) features, 330 nm absorption shoulder, and a short type 3 (T3) Cu-Cu distance, while the alternative resting form shows unusually small A(||) and high g(||) EPR features, lack of 330 nm absorption intensity, and a long T3 Cu-Cu distance. These different forms are evaluated with respect to activation for catalysis, and it is shown that the alternative resting form can only be activated by low-potential reduction, in contrast to the resting oxidized form which is activated via type 1 Cu at high potential. This difference in activity is correlated to differences in redox states of the two forms and highlights the requirement for efficient sequential reduction of resting MCOs for their involvement in catalysis.

  14. Urban forms, physical activity and body mass index: a cross-city examination using ISS Earth Observation photographs

    NASA Technical Reports Server (NTRS)

    Lin, Ge

    2005-01-01

    Johnson Space Center has archived thousands of astronauts acquired Earth images. Some spectacular images have been widely used in news media and in k-12 class room, but their potential utilizations in health promotion and disease prevention have relatively untapped. The project uses daytime ISS photographs to define city forms and links them to city or metropolitan level health data in a multicity context. Road connectivity, landuse mix and Shannon's information indices were used in the classification of photographs. In contrast to previous remote-sensing studies, which tend to focus on a single city or a portion of a city, this project utilized photographs of 39 U.S. cities. And in contrast to previous health-promotion studies on the built environment, which tend to rely on survey respondents' responses to evaluate road connectivity or mixed land use for a single study site, the project examined the built environments of multiple cities based on ISS photos. It was found that road connectivity and landuse mix were not statistically significant by themselves, but the composite measure of the Shannon index was significantly associated with physical activity, but not BMI. Consequently, leisure-time physical activity seems to be positively associated with the urban complexity scale. It was also concluded that unless they are planned or designed in advance, photographs taken by astronauts generally are not appropriate for a study of a single-site built environment nor are they appropriate for a study of infectious diseases at a local scale. To link urban built environment with city-wide health indicators, both the traditional nadir view and oblique views should be emphasized in future astronauts' earth observation photographs.

  15. In vitro and in vivo activities of the hydroxynaphthoquinone 566C80 against the cyst form of Toxoplasma gondii.

    PubMed Central

    Araujo, F G; Huskinson-Mark, J; Gutteridge, W E; Remington, J S

    1992-01-01

    The in vitro and in vivo activities of the hydroxynaphthoquinone 566C80 against the cyst form of Toxoplasma gondii were evaluated. In vitro treatment (100 micrograms of 566C80 per ml for 3 days) of cysts isolated from brains of mice infected for 1, 2, 3, 4, or 9 months resulted in loss of viability of the cysts and did not reveal any influence of the duration of in vivo infection on sensitivity to the drug. In vivo experiments to determine the effect of prolonged treatment with 200 mg of 566C80 per kg of body weight per day on cysts in brains of CBA/Ca mice infected with strain ME49 revealed a steady and significant decline in the numbers of cysts compared with the numbers in untreated controls. Histopathology of brains from control mice revealed inflammatory infiltrates around capillaries and in the parenchymas and meninges which were consistently less evident in the brains of treated mice. In addition, cysts were rarely observed in treated mice, whereas extensive inflammation and large numbers of cysts were found throughout the entire brain in control mice infected for the same period. The reduction in the numbers of cysts was evident as early as day 5 of treatment but was more marked at 8 weeks of treatment. The numbers of cysts in the brains of Swiss Webster mice infected for 3 or 6 months also significantly decreased following treatment for 15 or 30 days with the same dose of 566C80. Our results indicate that 566C80 has excellent activity against cysts of T. gondii both in vivo and in vitro and that sensitivity of the cysts to 566C80 is not affected by the duration of the infection in vivo. Images PMID:1605598

  16. 75 FR 5061 - Commission Information Collection Activities (FERC Form Nos. 6 and 6-Q); Comment Request; Extensions

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-01

    ... information reported on the annual FERC Form 6 and quarterly FERC Form 6-Q provides FERC, as well as customers... jurisdictional entities assists the Commission in its analysis of profitability, efficiency, risk and in...

  17. 76 FR 17426 - Agency Information Collection Activities: Application for Waiver of Passport and/or Visa (Form I...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-29

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  18. 78 FR 24429 - Agency Information Collection Activities: Inter-Agency Alien Witness and Informant Record, Form I...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-25

    ... Alien Witness and Informant Record, Form I-854; Extension, Without Change, of a Currently Approved... Collection. (2) Title of the Form/Collection: Inter-Agency Alien Witness and Informant Record. (3) Agency...: State, local or Tribal Government. Form I-854 is used by law enforcement agencies to bring...

  19. 77 FR 61776 - Agency Information Collection Activities: Petition for Alien Fiancé(e), Form I-129F, Revision of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-11

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    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-25

    ... Alien Witness and Informant Record, Form I-854, Extension Without Change, of a Currently Approved... Alien Witness and Informant Record. (3) Agency form number, if any, and the applicable component of the...: Primary: Individuals or Households. Form I- 854 is used by law enforcement agencies to bring...