Recombinant Collagen Engineered to Bind to Discoidin Domain Receptors Functions as a Receptor Inhibitor

A bacterial collagen-like protein Scl2 has been developed as a recombinant collagen model system to host human collagen ligand-binding sequences, with the goal of generating biomaterials with selective collagen bioactivities. Defined binding sites in human collagen for integrins, fibronectin, heparin, and MMP-1 have been introduced into the triple-helical domain of the bacterial collagen and led to the expected biological activities. The modular insertion of activities is extended here to the discoidin domain receptors (DDRs), which are collagen-activated receptor tyrosine kinases. Insertion of the DDR-binding sequence from human collagen III into bacterial collagen led to specific receptor binding. However, even at the highest testable concentrations, the construct was unable to stimulate DDR autophosphorylation. The recombinant collagen expressed in Escherichia coli does not contain hydroxyproline (Hyp), and complementary synthetic peptide studies showed that replacement of Hyp by Pro at the critical Gly-Val-Met-Gly-Phe-Hyp position decreased the DDR-binding affinity and consequently required a higher concentration for the induction of receptor activation. The ability of the recombinant bacterial collagen to bind the DDRs without inducing kinase activation suggested it could interfere with the interactions between animal collagen and the DDRs, and such an inhibitory role was confirmed in vitro and with a cell migration assay. This study illustrates that recombinant collagen can complement synthetic peptides in investigating structure-activity relationships, and this system has the potential for the introduction or inhibition of specific biological activities

Archaeal DNA Polymerase-B as a DNA Template Guardian: Links between Polymerases and Base/Alternative Excision Repair Enzymes in Handling the Deaminated Bases Uracil and Hypoxanthine

In Archaea repair of uracil and hypoxanthine, which arise by deamination of cytosine and adenine, respectively, is initiated by three enzymes: Uracil-DNA-glycosylase (UDG, which recognises uracil); Endonuclease V (EndoV, which recognises hypoxanthine); and Endonuclease Q (EndoQ), (which recognises both uracil and hypoxanthine). Two archaeal DNA polymerases, Pol-B and Pol-D, are inhibited by deaminated bases in template strands, a feature unique to this domain. Thus the three repair enzymes and the two polymerases show overlapping specificity for uracil and hypoxanthine. Here it is demonstrated that binding of Pol-D to primer-templates containing deaminated bases inhibits the activity of UDG, EndoV, and EndoQ. Similarly Pol-B almost completely turns off EndoQ, extending earlier work that demonstrated that Pol-B reduces catalysis by UDG and EndoV. Pol-B was observed to be a more potent inhibitor of the enzymes compared to Pol-D. Although Pol-D is directly inhibited by template strand uracil, the presence of Pol-B further suppresses any residual activity of Pol-D, to near-zero levels. The results are compatible with Pol-D acting as the replicative polymerase and Pol-B functioning primarily as a guardian preventing deaminated base-induced DNA mutations

Novel inhibition of archaeal family-D DNA polymerase by uracil.

International audienceArchaeal family-D DNA polymerase is inhibited by the presence of uracil in DNA template strands. When the enzyme encounters uracil, following three parameters change: DNA binding increases roughly 2-fold, the rate of polymerization slows by a factor of ≈ 5 and 3'-5' proof-reading exonuclease activity is stimulated by a factor of ≈ 2. Together these changes result in a significant decrease in polymerization activity and a reduction in net DNA synthesis. Pol D appears to interact with template strand uracil irrespective of its distance ahead of the replication fork. Polymerization does not stop at a defined location relative to uracil, rather a general decrease in DNA synthesis is observed. 'Trans' inhibition, the slowing of Pol D by uracil on a DNA strand not being replicated is also observed. It is proposed that Pol D is able to interact with uracil by looping out the single-stranded template, allowing simultaneous contact of both the base and the primer-template junction to give a polymerase-DNA complex with diminished extension ability

Early (<<0.3 day) R-band light curve of the optical afterglow of GRB030329

We observed the optical afterglow of the bright gamma-ray burst GRB030329 on the nights of 2003 March 29, using the Kiso observatory (the University of Tokyo) 1.05 m Schmidt telescope. Data were taken from March 29 13:21:26 UT to 17:43:16 (0.072 to 0.253 days after the burst), using an $Rc$-band filter. The obtained $Rc$-band light curve has been fitted successfully by a single power law function with decay index of $0.891\pm0.004$. These results remain unchanged when incorporating two early photometric data points at 0.065 and 0.073 days, reported by Price et al.(2003) using the SSO 40 inch telescope, and further including RTT150 data (Burenin et al. 2003) covering at about 0.3 days. Over the period of 0.065-0.285 days after the burst, any deviation from the power-law decay is smaller than $\pm$0.007 mag. The temporal structure reported by Uemura et al. (2003) does not show up in our $R$-band light curve.Comment: 9 pages, 2 figures, 1 table, accepted for publication in ApJ

Low energy high angular resolution neutral atom detection by means of micro-shuttering techniques: the BepiColombo SERENA/ELENA sensor

The neutral sensor ELENA (Emitted Low-Energy Neutral Atoms) for the ESA cornerstone BepiColombo mission to Mercury (in the SERENA instrument package) is a new kind of low energetic neutral atoms instrument, mostly devoted to sputtering emission from planetary surfaces, from E ~20 eV up to E~5 keV, within 1-D (2x76 deg). ELENA is a Time-of-Flight (TOF) system, based on oscillating shutter (operated at frequencies up to a 100 kHz) and mechanical gratings: the incoming neutral particles directly impinge upon the entrance with a definite timing (START) and arrive to a STOP detector after a flight path. After a brief dissertation on the achievable scientific objectives, this paper describes the instrument, with the new design techniques approached for the neutral particles identification and the nano-techniques used for designing and manufacturing the nano-structure shuttering core of the ELENA sensor. The expected count-rates, based on the Hermean environment features, are shortly presented and discussed. Such design technologies could be fruitfully exported to different applications for planetary exploration.Comment: 11 page

Development of High Ic Long REBCO Tapes with High Production Rate by PLD Method

AbstractWe have been developing long REBa2Cu3O7-δ coated conductors with high performance by the combination of the IBAD and the PLD methods. To realize the low production cost for REBa2Cu3O7-δ coated conductors, growth conditions were optimized for long tape fabrication in the “in-plume PLD method”. As a result, the Ic performance was confirmed with a high production rate under the high oxygen gas pressure and high laser energy density of > 800 mTorr and > 3J/cm2, respectively. We successfully fabricated a 35 m long GdBa2Cu3O7-δ coated conductor with high Ic value of 619 A/cm-w by the production rate of 30 m/h

Precise Control of Band Filling in NaxCoO2

Electronic properties of the sodium cobaltate NaxCoO2 are systematically studied through a precise control of band filling. Resistivity, magnetic susceptibility and specific heat measurements are carried out on a series of high-quality polycrystalline samples prepared at 200 C with Na content in a wide range of 0.35 =< x =< 0.70. It is found that dramatic changes in electronic properties take place at a critical Na concentration x* that lies between 0.58 and 0.59, which separates a Pauli paramagnetic and a Curie-Weiss metals. It is suggested that at x* the Fermi level touches the bottom of the a1g band at the gamma point, leading to a crucial change in the density of states across x* and the emergence of a small electron pocket around the gamma point for x > x*.Comment: 4 pages, 5 figures, submitted to J. Phys. Soc. Jp

Structural and biochemical characterization of the novel CTXM-151 extended-spectrum β-lactamase and its inhibition by avibactam

The diazabicyclooctane (DBO) inhibitor avibactam (AVI) reversibly inactivates most serine β-lactamases, including the CTX-M β-lactamases. Currently, more than 230 unique CTX-M members distributed in five clusters with less than 5% amino acid sequence divergence within each group have been described. Recently, a variant named CTX-M-151 was isolated from a Salmonella enterica subsp. enterica serovar Choleraesuis strain in Japan. This variant possesses a low degree of amino acid identity with the other CTX-Ms (63.2% to 69.7% with respect to the mature proteins), and thus it may represent a new subgroup within the family. CTX-M-151 hydrolyzes ceftriaxone better than ceftazidime (kcat/Km values 6,000-fold higher), as observed with CTX-Ms. CTX-M-151 is well inhibited by mechanism-based inhibitors like clavulanic acid (inactivation rate [kinact]/inhibition constant [Ki] = 0.15μM-1 · s-1). For AVI, the apparent inhibition constant (Ki app), 0.4mM, was comparable to that of KPC-2; the acylation rate (k2/K) (37,000 M-1 · s-1) was lower than that for CTX-M-15, while the deacylation rate (koff) (0.0015 s21) was 2- to 14-fold higher than those of other class A β-lactamases. The structure of the CTX-M-151/AVI complex (1.32 Å) reveals that AVI adopts a chair conformation with hydrogen bonds between the AVI carbamate and Ser70 and Ser237 at the oxyanion hole. Upon acylation, the side chain of Lys73 points toward Ser130, which is associated with the protonation of Glu166, supporting the role of Lys73 in the proton relay pathway and Glu166 as the general base in deacylation. To our knowledge, this is the first chromosomally encoded CTX-M in Salmonella Choleraesuis that shows similar hydrolytic preference toward cefotaxime (CTX) and ceftriaxone (CRO) to that toward ceftazidime (CAZ).Fil: Ghiglione, Barbara. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquimica. Instituto de Investigaciones En Bacteriologia y Virologia Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Rodríguez, María Margarita. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquimica. Instituto de Investigaciones En Bacteriologia y Virologia Molecular; ArgentinaFil: Brunetti, Florencia Lourdes. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquimica. Instituto de Investigaciones En Bacteriologia y Virologia Molecular; ArgentinaFil: Papp Wallace, Krisztina M.. Case Western Reserve University; Estados UnidosFil: Yoshizumi, Ayumi. Toho University; JapónFil: Ishii, Yoshikazu. Toho University; JapónFil: Bonomo, Robert A.. Case Western Reserve University; Estados UnidosFil: Gutkind, Gabriel Osvaldo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquimica. Instituto de Investigaciones En Bacteriologia y Virologia Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Klinke, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Power, Pablo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquimica. Instituto de Investigaciones En Bacteriologia y Virologia Molecular; Argentin

Conventional and CT angiography in children: dosimetry and dose comparisons

Tremendous advances have been made in imaging in children with both congenital and acquired heart disease. These include technical advances in cardiac catheterization and conventional angiography, especially with advancements in interventional procedures, as well as noninvasive imaging with MR and CT angiography. With rapid advances in multidetector CT (MDCT) technology, most recently 64-detector array systems (64-slice MDCT), have come a number of advantages over MR. However, both conventional and CT angiography impart radiation dose to children. Although the presence of radiation exposure to children has long been recognized, it is apparent that our ability to assess this dose, particularly in light of the rapid advancements, has been limited. Traditional methods of dosimetry for both conventional and CT angiography are somewhat cumbersome or involve a potential for substantial uncertainty. Recent developments in dosimetry, including metal oxide semiconductor field effect transistors (MOSFET) and the availability of anthropomorphic, tissue-equivalent phantoms have provided new opportunities for dosimetric assessments. Recent work with this technology in state-of-the-art cardiac angiography suites as well as with MDCT have offered direct comparisons of doses in infants and children undergoing diagnostic cardiac evaluation. It is with these dose data that assessment of risks, and ultimately the assessment of risk-benefit, can be better achieved